Table of Content

30 December 2024, Volume 42 Issue 6

Previous Issue    Next Issue

SPECIAL REPORT

National surveillance on soil-transmitted helminth infections in China, 2021

ZHAO Luyuan, HUANG Jilei, ZHOU Changhai, ZHU Tingjun, ZHU Huihui, ZHOU Xiaonong, LI Shizhu, QIAN Menbao

2024, 42(6):  687-693.  doi:10.12140/j.issn.1000-7423.2024.06.001

Asbtract ( )   HTML ( )   PDF (640KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the prevalence and epidemiological characteristics of soil-transmitted helminth (STH) infections in China in 2021, so as to provide data support for improving soil-transmitted helminthiasis control strategies in high-prevalence areas and implementing transmission control and interruption of soil-transmitted helminthiasis in low-prevalence areas. Methods National surveillance on soil-transmitted helminthiasis was performed in 412 surveillance counties across 31 provinces (municipalities and autonomous regions) in China in 2021. Each surveillance county was divided into eastern, western, southern, northern, and central parts according to geographical locations, and one administrative village was sampled from one township in each part. Then, 200 permanent residents over 3 years old were sampled from each administrative village, and at least 1 000 residents of different age groups were recruited in each surveillance county. Subjects’ stool samples were collected for detection of STH eggs using a modified Kato-Katz thick smear method (two smears from one stool). In addition, 5 households were randomly sampled from each administrative village, and soil samples were collected from their farmlands or vegetable gardens to detect hookworm larvae and Ascaris lumbricoides eggs. Comparisons of proportions were done with the Chi-square test. Results A total of 424 306 residents were investigated in 412 soil-transmitted helminthiasis surveillance counties across China in 2021, and the overall prevalence of STH infections was 0.88% (3 730/424 306), with 0.67% (2 845/424 306) of hookworm infection, 0.11% (461/424 306) of A. lumbricoides infection and 0.12% (526/424 306) of Trichuris trichura infection, respectively. The highest prevalence of STH infections was demonstrated in Yunnan Province (5.74%, 917/15 967), the highest prevalence of hookworm infection was also in Yunnan Province (4.93%, 787/15 967), the highest prevalence of A. lumbricoides infection was in Qinghai Province (0.52%, 32/6 106), and the highest prevalence of T. trichura infection was in Hainan Province (1.10%, 46/4 184). The prevalence of STH infections was higher in females (0.94%, 2 052/217 245) than in males (0.81%, 1 678/207 061) (χ² = 21.90, P < 0.01), and the highest prevalence of STH infections was shown among residents aged over 60 years (1.62%, 1 921/118 413) (χ² = 1 175.93, P < 0.01). The proportions of light-intensity infections in hookworm, A. lumbricoides and T. trichura were 92.16% (2 622/2 845), 90.46% (417/461) and 93.54% (492/526), and the proportions of moderate infections were 4.04% (115/2 845), 9.54% (44/461) and 6.08% (32/526). There were 108 residents detected with heavy-intensity hookworm infections (3.80%, 108/2 845), and 2 participants with heavy-intensity T. trichura infections (0.38%, 2/526); however, no heavy-intensity A. lumbricoides infection was found. A. lumbricoides eggs were detected in 73 out of 2 475 households of soil samples (2.95%, 73/2 475), and hookworm larvae were identified in 58 households of soil samples (2.34%, 58/2 475). Both A. lumbricoides eggs and hookworm larvae were detected in soil samples from farmlands and vegetable gardens. In addition, the detection rates of Necator americanus and Ancylostoma duodenale larvae were 1.86% (46/2 475) and 0.40% (10/2 475), respectively. Conclusion The prevalence of STH infections was reduced to less than 1% in national surveillance counties of China in 2021; however, it varied greatly in regions. More attention is required to be paid to residents at ages of over 60 years and females. Precise control strategies tailored to local circumstances are recommended, and initiation of transmission control and interruption of soil-transmitted helminthiasis is encouraged in low-prevalence areas.

ORIGINAL ARTICLES

Serological epidemic characteristics of transient population at national schistosomiasis surveillance sites of China, 2020-2022

GUO Suying, LI Yinlong, LI Shizhen, DANG Hui, CAO Chunli, XU Jing, LI Shizhu

2024, 42(6):  694-700.  doi:10.12140/j.issn.1000-7423.2024.06.002

Asbtract ( )   HTML ( )   PDF (2455KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the seroepidemiological characteristics in transient population and the moving direction of the individuals with positive serum anti-schistosome antibody at national schistosomiasis surveillance sites of China during 2020-2022, and to provide a scientific basis for formulating surveillance program for schistosomiasis in the transient population. Methods Data of the national population-based schistosomiasis surveillance and individual cases recorded from transient population during 2020-2022 were collected from the Information System for Infectious Diseases Prevention and Control and the Information System for Parasitic Disease Prevention and Control. The data include demographic characteristics of sex, age, occupation and the counties of residence registered, and the people migrating information regarding the locations of moving in or moving out from, as well as the results of serological and pathogenic examinations for schistosomiasis. Descriptive epidemiological methods were employed to analyze the spatial, temporal, and demographic distribution of the serum anti-schistosome level among the transient population. Chi-square analysis was used to compare the differences in the positive rates of serum antibodies among the groups of sex, age, and occupation. Partitions of χ² method was used for multiple comparisons of the positive rates. Sankey diagram was used to describe the moving direction of the individuals with positives serum antibodies. Results A total of 302 460 person-time among the transient population was surveyed from 455 surveillance counties in 13 provinces (or autonomous regions or municipalities) in China from 2020 to 2022, and 1 199 was found positive with serum antibodies, and one was positive with fecal examination. During 2020-2022, the positive rates of serum antibodies among the transient population were 0.47% (450/96 596), 0.44% (442/101 558), and 0.29% (307/104 306), respectively. The rates in 2020 and 2021 were both higher than the rate in 2022 (χ² = 39.310, 28.178; both P < 0.01). During 2020-2022, the highest positive rates of serum antibodies were detected in Jiangxi, showing 1.24% (91/7 316), 1.33% (112/8 412), and 0.65% (56/8 648), respectively. The positive rates of serum antibodies were 0.48% (277/57 550), 0.44% (260/59 458), and 0.31% (191/62 237) in males and 0.44% (173/39 046), 0.43% (182/42 100), and 0.28% (116/42 069) in females, respectively. No statistically significant differences were observed between sex groups in each year (χ² = 0.734, 0.014, 0.830; all P > 0.05). The positive rates of serum antibodies were 0.82% (225/27 561), 0.78% (226/28 928) and 0.53% (158/29 661) in farmers, 0.86% (20/2 333), 0.75% (16/2 141) and 0.91% (23/2 523) in boatmen/fishermen, and 0.31% (205/66 702), 0.28% (200/70 489) and 0.17% (126/72 122) in other occupation group, respectively. The positive rates of serum antibodies were higher in the farmers and boatmen/fishermen groups compared to the other occupation group (χ² = 111.287, 118.991, 96.799; 20.985, 15.060, 66.450; all P < 0.01). The positive rates of serum antibodies in group of 6-15 years were the highest (1.00%, 16/1 597) in 2020, the second highest (0.70%, 10/1 436) in 2021, and the lowest (0.05%, 1/1 836) in 2022, respectively. The differences among different age groups were statistically significant for each year (χ² = 41.282, 56.515, 54.425; all P < 0.01). Among the seropositive individuals who provided complete information on their migration regions, higher proportions of emigration location were seen from Sichuan (21.86%, 256/1 171), Jiangxi (21.18%, 248/1 171), and Hubei (18.02%，211/1 171), and the highest two destinations were Jiangxi (18.87%, 221/1 171) and Sichuan (18.87%, 221/1 171). Among the seropositive individuals who provided complete migration information, higher proportions of emigration location were seen from Sichuan (21.86%, 256/1 171), Jiangxi (21.18%, 248/1 171), and Hubei (18.02%, 211/1 171). The higher proportions of immigration location were seen in Jiangxi (18.87%, 221/1 171) and Sichuan (18.87%, 221/1 171); the proportion of immigration location within or between non-eliminated provinces was 73.87% (865/1 171), and the proportion of migrants moving between non-eliminated and eliminated provinces was 19.98% (234/1 171). Forty-six individuals with positive serum antibodies migrated from non-endemic areas to endemic areas. Conclusion According to the survey of the national surveillance sites, the prevalence of schistosomiasis in transient population in China remained at a very low level from 2020 to 2022. It is imperative to strengthen the surveillance for the transient population in Jiangxi, younger age group and the migrants moving in or out from non-elimination areas, as well as the proper management of people with past infections.

Epidemiology and determinants of Clonorchis sinensis infection in Binyang County, Guangxi in 2022

ZHU Yuying, ZHAO Jiaguang, ZHOU Changhai, ZHU Tingjun, HUANG Jilei, MENG Jun, JIANG Zhihua, ZHOU Xiaonong, LI Shizhu, QIAN Menbao

2024, 42(6):  701-714.  doi:10.12140/j.issn.1000-7423.2024.06.003

Asbtract ( )   HTML ( )   PDF (2137KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the epidemiological characteristics of, and knowledge, attitude, and practice towards Clonorchis sinensis infections in humans in Binyang County, Guangxi in 2022, for providing insights into formulation of targeted control measures. Methods Binyang County in Guangxi was geographically divided into five regions (eastern, western, southern, northern, and central) in 2022, and one township was sampled from each geographical region, followed by one administrative village (community) sampled from each township. Two hundred residents at ages of over 3 years were sampled from each administrative village (community) using a cluster sampling method, and participants’ stool specimens were sampled for detection of C. sinensis eggs using a modified Kato-Katz thick smear method (two smears from one stool sample). One questionnaire survey was performed in each household, and 40 residents were sampled from each village for individual questionnaire surveys on knowledge, attitude, and practice towards clonorchiasis control. The differences in the awareness of clonorchiasis control knowledge, practice towards consumption of raw freshwater fish, and attitude to consume raw freshwater fish were compared among different participants with Chi-square test or Fisher’s exact test. In addition, univariate and multivariate logistic regression models were employed to identify factors affecting the prevalence of C. sinensis infections in humans, practice of consuming raw freshwater fish, and attitude towards consumption of raw freshwater fish in future among participants. Results A total of 1 007 residents received stool examinations, and 372 household questionnaire surveys and 202 individual questionnaire surveys were completed, including 175 individuals completing both stool examinations and questionnaire surveys. The overall prevalence of C. sinensis infections was 15.49% [95% confidential interval (CI): (13.25%, 17.73%)] among participants, with the highest prevalence in Silong Township [22.00%; 95% CI: (15.84%, 27.38%)] (P < 0.05), and the prevalence was higher in males [26.32%; 95% CI: (22.27%, 30.06%)] than females [5.07%; 95% CI: (3.16%, 6.97%)] (P < 0.05). The prevalence of C. sinensis infections in humans raised and then decreased following the increase of ages, with a peak at ages of 45 to 59 years [24.31%; 95% CI: (19.01%, 29.61%)] (P < 0.05), and the highest prevalence was found among participants with an educational level of junior high school [23.77%; 95% CI: (19.89%, 27.64%)] (P < 0.05). Among the 372 households surveyed, the proportions of households preparing raw freshwater fish less than once, once to 10 times, 11 to 50 times and more than 50 times annually were 4.03% (15/372), 21.77% (81/372), 15.06% (56/372) and 8.60% (32/372), respectively. Notably, Binzhou Township recorded the highest proportion of households preparing raw freshwater fish once to 10 times annually (83.64%, 46/55), and Xinxu Township recorded the highest proportion of households preparing raw freshwater fish 11 to 50 times annually (50.47%, 54/107) and more than 50 times annually (29.91%, 32/107), respectively (P < 0.05). The highest proportion of households that had purchased raw freshwater fish from outside was found in Silong Township (35.09%, 20/57) (P < 0.05), and the highest proportion of households that had presented raw freshwater fish to their neighbors was seen in Binzhou Township (51.81%, 43/55) (P < 0.05), while both the highest proportion of households that had received raw freshwater fish from neighbors (99.07%, 106/107) and did not separate raw and cooked chopping boards in daily life (98.08%, 102/104) was found in Xinxu Township (P < 0.05). The awareness of hearing of clonorchiasis, knowing the transmission route of clonorchiasis and knowing the harm of clonorchiasis was 54.46% (110/202), 50.00% (101/202) and 38.12% (77/202) among all participants, with 63.00% (63/100), 59.00% (59/100), and 46.00% (46/100) in males and 46.08% (47/102), 41.18% (42/102), and 30.39% (31/102) in females, respectively (P < 0.05), and the awareness was higher in Gula Township (92.50%, 37/40; 92.50%, 37/40; 90.00%, 36/40), among the elderly at ages of 60 years and older (71.43%, 45/63; 63.49%, 40/63; 50.79%, 32/63), Han ethnic group (58.29%, 102/175; 54.29%, 95/175; 41.14%, 72/175), and participants with an educational level of senior high school and above (87.50%, 14/16; 81.25%, 13/16; 62.50%, 10/16) (P < 0.05). There were 26.73% (54/202) of respondents reporting ingestion of raw freshwater fish, with a higher percentage in Silong Township (45.00%, 18/40) (P < 0.05). The percentage of consuming raw freshwater fish was higher in males (45.00%, 45/100) than females (8.82%, 9/102) (P < 0.05), and the highest proportion of consuming raw freshwater fish was seen among participants at ages of 30 to 44 years (42.31%, 11/26) (P < 0.05). There were 80.69% (163/202) of respondents that were willing to stop consuming raw freshwater fish in future, with a higher proportion in females (97.06%, 99/102) than males (64.00%, 64/100) (P < 0.05). Univariate logistic regression analysis showed that C. sinensis infection was associated with gender, age, awareness of clonorchiasis, frequency of preparing and consuming raw freshwater fish and way of acquiring raw freshwater fish (P < 0.05), and the practice of consuming raw freshwater fish was associated with gender, awareness of clonorchiasis, understanding routes of C. sinensis infection, having experiences of preparing raw freshwater fish, having experiences of purchasing raw freshwater fish from outside, and having experiences of presenting or receiving raw freshwater fish (P < 0.05), while factors affecting the attitude towards consumption of raw freshwater fish in future included gender, age, positive fecal test, frequency of consuming raw freshwater fish, and having experiences of receiving raw freshwater fish from neighbors (P < 0.05). Multivariate logistic regression analysis revealed that the prevalence of C. sinensis infections was 30.01 [95% CI: (7.56, 152.04)] times among respondents consuming raw freshwater fish once to 5 times annually and 81.40 [95% CI: (7.78, 287.37)] times than among those consuming raw freshwater fish more than 5 times annually relative to individuals without ingestion of raw freshwater fish, and the prevalence was 15.18 [95% CI: (2.15, 123.25)] times higher among respondents that had received raw freshwater fish from their neighbors than among those who had not. The proportion of consuming raw freshwater fish was 17.58 [95% CI: (6.21, 61.88)] times in males than females, and the proportion of consuming raw freshwater fish was 15.75 [95% CI: (1.40, 185.99)] times among respondents that knew clonorchiasis than among those who did not, while the percentages of continuing to consume raw freshwater fish were 3.66 [95% CI: (1.85, 4.32)] times among respondents with consumption of raw freshwater fish once to five times annually and 3.55 [95% CI: (1.17, 3.03)] times among those with consumption of raw freshwater fish more than five times annually than among individuals without consumption of raw freshwater fish. Conclusion The prevalence of C. sinensis infection is high in Binyang County, Guangxi, notably among adult males, and the special dietary habit of consuming raw freshwater fish is the major cause of C. sinensis infections among human. Intensified health education and changes of the behavior of consuming raw freshwater fish are required to achieve sustainable clonorchiasis control.

Prevalence of Fasciola infections and molecular identification of Fasciola in livestock in Dali Bai Autonomous Prefecture, Yunnan Province

LI Shiyun, LI Cihuai, CHEN Feng, HAO Mingming, LI Xindi, ZHAO Yinjiao, ZHANG Dongqi, ZHANG Quanfu, LI Qincui, ZHANG Jianping, CHEN Shaorong, LUO Tianpeng, LIU Yuhua

2024, 42(6):  715-720.  doi:10.12140/j.issn.1000-7423.2024.06.004

Asbtract ( )   HTML ( )   PDF (1062KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the prevalence of Fasciola infections and to characterize the Fasciola species in livestock in Dali Bai Autonomous Prefecture. Methods Cattle, sheep, horse and pig stool samples were collected from 12 counties (city) in Dali Bai Autonomous Prefecture during the period from August 2023 to March 2024 for detection of Fasciola eggs. Parasite eggs were sampled from three Fasciola-infected stool specimens in each county, and photographed and measured with a microscopic imaging system. Then, genomic DNA was extracted from Fasciola eggs, and the ribosomal internal transcribed spacer (ITS) gene was amplified using a PCR assay for characterization of Fasciola species. The sequence of the ITS gene was aligned using the BLAST program in NCBI, and a neighbor-joining phylogenetic tree of Fasciola spp. was built based on the ITS gene sequence using the software MEGA 11.0. All statistical analyses were performed using the software SPSS 19.0, and differences in the prevalence of Fasciola infections were compared with chi-square test. Results A total of 1 302 livestock stool samples were collected, including 1 228 cattle stool samples (94.3%), 45 sheep stool samples (3.5%), 18 horse stool samples (1.4%), and 11 pig stool samples (0.8%), and there were 212 stool samples detected with suspected Fasciola eggs (16.3%). The detection of Fasciola eggs was 17.2% in cattle stool samples (211/1 228) and 2.2% in sheep stool samples (1/45), and no Fasciola eggs were detected in horse or pig stool samples. The highest detection of Fasciola eggs was seen in livestock stool samples from Weishan County (45.0%, 49/109), followed by from Eryuan County (42.7%, 44/103), Yunlong County (28.2%, 29/103), Midu County (25.4%, 30/118), Binchuan County (18.0%, 18/100), Heqing County (16.5%, 17/103), Xiangyun County (8.7%, 9/103), Jianchuan County (6.9%, 8/116), Nanjian County (4.7%, 5/107), Yangbi County (2.2%, 3/137), and no Fasciola eggs were detected in livestock stool samples from Dali City or Yongping County. There was a region-specific prevalence of Fasciola infections in livestock in Dali Bai Autonomous Prefecture (χ² = 222.975, P < 0.05), and the prevalence of Fasciola infections was higher in female livestock (19.0%, 165/873) than in males (11.0%, 47/429) (χ² = 13.319, P < 0.05). The eggs of F. gigantica and F. hepatica measured (182.30 ± 4.35) μm × (97.80 ± 5.11) μm and (149.49 ± 5.90) μm × (80.46 ± 4.37) μm, respectively. A total of 7 ITS gene sequences were obtained, including 2 from F. hepatica and 5 from F. gigantica, and these 7 sequences had been uploaded to the National Microbiology Data Center (NMDC), with NMDC numbers of NMDCN0005QTQ (sequence number: Binchuan County A1), NMDCN0005TN (sequence number: Weishan County A32), NMDCN0005QTO (sequence number: Jianchuan County A70), NMDCN0005ELF (sequence number: Yangbi County A19), NMDCN0005QTR (sequence number: Heqing County A7), NMDCN0005ELH (sequence number: Midu County A35) and NMDCN0005QTP (sequence number: Xiangyun County A18). Four sequences numbered Binchuan County A1, Weishan County A32, Jianchuan County A70 and Heqing County A7 had 100% sequence homology to those from F. gigantica (GenBank accession numbers: JF496711, OQ064778, MN608171 and MN97009), and the sequence numbered Xiangyun County A18 had 93% sequence homology to that from F. gigantica (GenBank accession number: MN97009), while two sequences numbered Yangbi County A19 and Weishan County A35 had 100% sequence homology to those from F. hepatica (GenBank accession number: JF432071 and JF496716). Phylogenetic analysis showed that the sequence numbered Yangbi A19 and sequences of F. hepatica from Australia (GenBank accession number: MN970007) and Iran (GenBank accession number: JF432071) were clustered into the same clade, and the sequence numbered Midu A35 and that of F. hepatica from Dali, China (GenBank accession number: JF496716) was clustered into the same clade, indicating a close genetic relationship. In addition, the remaining five sequences and those of F. gigantica from Dehong, China (GenBank accession number: JF496711), Bangladesh (GenBank accession number: OQ064778), Nigeria (GenBank accession number: MN608171), and Vietnam (GenBank accession numbers: MN970009 and MN970008) were clustered into the same clade, indicating a close genetic relationship. Conclusions The prevalence of Fasciola infections is high in livestock Dali Bai Autonomous Prefecture, Yunnan Province, and F. gigantica is the predominant species of Fasciola.

Epidemiological characteristics of visceral leishmaniasis in Linfen City from 2013 to 2022

SHAO Hongli, FENG Meijuan, HUANG Li, SHI Lin’ai, LIANG Hongyun, ZHANG Jian, ZHANG Chonghui

2024, 42(6):  721-725.  doi:10.12140/j.issn.1000-7423.2024.06.005

Asbtract ( )   HTML ( )   PDF (654KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the epidemiological characteristics of visceral leishmaniasis in Linfen City, Shanxi Province from 2013 to 2022, so as to provide insights into prevention and control of visceral leishmaniasis. Methods All data pertaining to reported visceral leishmaniasis cases in Linfen City, Shanxi Province from January 1, 2013 to December 31, 2022 were collected from the Infectious Diseases Report Information Management System of China Disease Prevention and Control Information System. The temporal, regional, age and occupation distributions of reported visceral leishmaniasis cases were analyzed using a descriptive epidemiological method. In addition, the sandfly density was monitored using sandfly light traps in Hongdong County, Jixian County and Huozhou City in July 2023, and the sandfly species was identified. Results A total of 78 visceral leishmaniasis cases were reported in Linfen City from 2013 to 2022, with no deaths, and the annual average incidence was 0.199 6/10⁵, which appeared a tendency towards a rise over years (χ² = 78.000, P < 0.01). Among them, a total of 3 cases were reported from 2013 to 2016, and the number has increased year by year since 2017. In 2021 and 2022, there were 19 and 28 cases respectively. Visceral leishmaniasis cases were reported in each month, with the highest number in April (14 cases, 17.95% of total cases), and the lowest number in October (1 case). Local cases of visceral leishmaniasis were reported in 10 counties (cities, districts) of Linfen City, and the three counties with the highest numbers of local cases included Xiangfen County (20 cases), Quwo County (20 cases), and Xiangning County (18 cases). The number of counties reporting cases has increased from 1 in 2013 to 8 in 2022. Among the 78 reported visceral leishmaniasis cases, there were 57 men and 21 women, with a male to female ratio of 2.71∶1, and the reported cases were predominantly aged 0 to 9 years (23 cases) and 60 to 69 years (19 cases), with a significant difference in the age distribution (χ² = 20.919, P < 0.01). Farmers (41 cases) and diaspora children (22 cases) were predominant occupations, and there were 42 cases (53.85%) diagnosed in other provinces (municipalities), and 36 cases (46.15%) diagnosed in Linfen City. A total of 2 299 female sandflies were captured and all were identified as Phlebotomus chinensis, with an average density of 1.58 sandflies/(lamp•night). Conclusion The epidemics of visceral leishmaniasis have appeared a tendency towards a rise in Linfen City since 2017, with expansion of epidemic areas and severe epidemics. Improved control awareness and intensified control efforts are recommended to control the transmission of visceral leishmaniasis.

Effect of harmine combined with rad51 inhibitor on the Echinococcus granulosus larvae and its DNA damage in vitro

GONG Yuehong, PAN Meichi, SUN Jiajia, MA Ruijia, WUSMANJIANG Aimaiti, ZHAO Yicong, ZHAO Jun, WANG Jianhua

2024, 42(6):  726-736.  doi:10.12140/j.issn.1000-7423.2024.06.006

Asbtract ( )   HTML ( )   PDF (4492KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the effect of harmine combined with rad51 inhibitor (RI-1) on Echinococcus granulosus in vitro and the level of DNA damage. Methods E. granulosus from the cysts of sheep liver were collected and randomly divided into groups. Each group contains a mixture of harmine (0, 20, 30, 40, 50, 100 μg/ml) and RI-1 (0, 5, 10, 20, 40, 80 μg/ml), cultured for 5 days, and stained with 0.1% methylene blue to calculate the survival rate. The group which is close to the half inhibitory concentration of harmine against E. granulosus was selected as the fixed concentration. The groups were treated against E. granulosus with a mixture of harmine and RI-1 in ratios of 40∶5, 40∶10 and 40∶20, respectively. The optimal synergistic ratio was screened. Set up a mixed drug group of harmine and RI-1, a harmine group and RI-1 group acting alone at the same concentration, and set up a positive control group (adding albendazole sulfoxide) and a blank control group. Observe the morphology of E. granulosus and vesicles after 3 days of intervention. DCFH-DA method was used to detect the reactive oxygen species content of E. granulosus after intervention in each group. Each group underwent comet assay (the positive control was doxorubicin), and the Olive tail moment was analyzed and measured. The reagent kit is used to detect the oxidative stress level and Caspase-3 content of E. granulosus after drug intervention, as well as the levels of adenosine triphosphate (ATP) and lactate dehydrogenase (LDH). qRT-PCR reaction is used to detect the mRNA expression levels of DNA damage repair related genes. Results The survival rates of E. granulosus after 5 days of intervention with 0, 20, 30, 40, 50 and 100 μg/ml harmine were (94.67 ± 1.25)%, (83.00 ± 3.27)%, (64.67 ± 3.86)%, (55.33 ± 3.40)%, (29.67 ± 2.49)%, and (9.33 ± 1.70)%, respectively. The survival rates of E. granulosus after 5 days of intervention with 0, 5, 10, 20, 40 and 80 μg/ml RI were (94.33 ± 0.94)%, (70.00 ± 2.94)%, (55.67 ± 3.86)%, and (30.00 ± 1.70)%, 5.09%, (2.67 ± 1.25)% and 0, respectively. After 3 days of intervention with a mixed drug ratio of 40 μg/ml∶10 μg/ml as the optimal synergistic ratio for E. granulosus, the survival rate of the mixed drug group was (48.67 ± 2.62)%, which was lower than that of the blank control group [(96.33 ± 1.25)%], positive control group [(91.67 ± 1.25)%], and harmine group [(72.67 ± 4.50)%] (F = 78.85, P < 0.01; F = 437.92, P < 0.05; F = 42.49, P < 0.01). The structure of the mixed drug group’s worm body collapsed, with severe morphological damage, significant shrinkage of the worm body, and insect like changes in the epidermal layer. The average fluorescence intensity of reactive oxygen species in the mixed drug group (45.71 ± 1.52) was enhanced compared with the blank control group (3.40 ± 0.55), the positive control group (12.98 ± 0.67), and the dehydroberberine group (39.79 ± 1.11) (F = 339.795, 773.96, 76.99, all P < 0.01). The Olive tail moment value of the mixed drug group (17.73 ± 1.11) was higher than that of the blank control group (0.03 ± 0.02), positive control group (6.46 ± 0.56), and dehydrocamel base group (8.80 ± 0.61) (F = 230.32, 165.59, 151.44, all P < 0.01). The SOD content in the mixed drug group was (0.71 ± 0.07) U/mg, which was lower than that in the blank control group [(2.32 ± 0.24) U/mg] and the dehydroberberine group [(1.26 ± 0.12) U/mg] (F = 39.57, 15.74, both P < 0.01). The MDA content in the mixed drug group was (0.62 ± 0.03) mmol/mg, which was lower than that in the blank control group [(0.82 ± 0.05) mmol/mg] and the dehydroberberine group [(0.70 ± 0.03) mmol/mg] (F = 22.78, 28.37, P < 0.01). The level of Caspase-3 in the mixed drug group was (1.82 ± 0.05) U/mg, which was higher than that in the blank control group [(0.70 ± 0.02) U/mg] and the dehydroberberine group [(1.36 ± 0.19) U/mg] (F = 26.24, P < 0.01; F = 7.52, P < 0.05). The mitochondrial ATP level of E. granulosus in the mixed drug group was (112.21 ± 2.72) µmol/ml, which was lower than that of the blank control group [(157.50 ± 5.23) µmol/ml] and the dehydrocoumarin group [(123.10 ± 4.73) µmol/ml] (F = 60.73, P < 0.01; F = 67.42, P < 0.05). The release of LDH from E. granulosus in the mixed drug group was (1.83 ± 0.09) mU, which was higher than that in the blank control group [(0.70 ± 0.05) mU] (F = 146.79, P < 0.01), but there was no statistically significant difference compared with the dehydrocoumarin group [(1.51 ± 0.05) mU] (F = 62.78, P > 0.05). The relative expression levels of h2ax, rad51, brca1 and mrel1 genes in the mixed drug group of E. granulosus were 2.34 ± 0.15, 1.19 ± 0.14, 1.35 ± 0.08, and 1.43 ± 0.10, respectively, which were upregulated compared with the blank control group (all 1.00 ± 0.00) (F = 23.79, P < 0.01; F = 8.96, P < 0.05; F = 30.75, P < 0.05; F = 39.81, P < 0.01). Conclusion The combination of harmine and rad51 inhibitor RI-1 can enhance the in vitro efficacy of harmine against E. granulosus, while also increasing DNA damage in E. granulosus.

Research on the application of 3D visualization-based technology to guide surgical plan for hepatic alveolar echinococcosis

LI Zhenwei, WANG Cheng, WANG Zhixin, LIU Jinming, ZHAO Qian, WANG Haijiu, XIE Zhi

2024, 42(6):  737-743.  doi:10.12140/j.issn.1000-7423.2024.06.007

Asbtract ( )   HTML ( )   PDF (2742KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective Exploring the innovative application of 3D visualization technology in assisting the design of surgical treatment plans for hepatic alveolar echinococcosis (HAE). Methods Fifty HAE patients who underwent preoperative examination with 3D visualization technology at the Affiliated Hospital of Qinghai University from January 2020 to March 2023 were enrolled to participate the research. The patient’s raw data was transmitted to a human 3D visualization virtual surgical system to process 3D visual reconstruction modeling for determining the whole liver volume, planned liver resection volume, the liver volume to be reserved, and the hydatid lesion volume, thereby,the residual liver ratio was calculated, and compared with what actually seen in the operation to evaluate the application value of 3D visualization technology for HAE. Another 50 patients with HAE who did not undergo 3D visualization technology during the same period were selected as the control group. The control group received 2D enhanced CT imaging and precise liver resection. Compare the general and perioperative data between the two groups were compared and the generalized estimating equation (GEE) model was used to analyze the influencing factors on patients’ postoperative prognosis. Results There was no statistically significant difference in general information such as age, gender, body mass index (BMI), resection site, lesion diameter, excised liver volume, remaining liver volume, as well as differences in CT features and MRI imaging data between two groups of patients (P > 0.05). Compared with the control group, the study group had shorter surgical time [(228.56 ± 29.47) min], pringle method total liver block time [(26.65 ± 4.78) min], less intraoperative blood loss [(435.14 ± 98.76) ml], shorter hospital stay [(15.27 ± 2.13) days], lower hospitalization costs [(4.58 ± 0.79) yuan], and higher satisfaction with doctor-patient communication (89.65 ± 9.23) (t = 3.856, 3.561, 6.069, 3.561, 5.400, 3.812; all P < 0.05). The reconstructed 3D model can be rotated, scaled, and translated, and can also hide or render semi-transparent structures such as liver parenchyma, veins, and arteries, providing a clear and intuitive understanding of the spatial relationships between important anatomical structures and lesions in the liver. The average volume of planned liver resection predicted by 3D visualization technology is 2 154.43 ml, the average volume of reserved liver is 1 203.27 ml, and the average residual liver ratio is 41.22%. The average volume of liver resection during the operation was 2 138.75 ml, and the average volume of remaining liver was 1 193.46 ml. There was no statistically significant difference (t = 1.135, P > 0.05) between the estimated and actual liver volumes obtained through 3D visualization reconstruction. The GEE analysis results showed that the invasion of hepatic Echinococcus multilocularis edge infiltration and 3D visualization reconstruction significantly affected the prognosis of patients (χ² = 7.890, 3.876; P < 0.05). Conclusion Three dimensional visualization technology can be used to develop a rational liver resection plan for HAE patients before surgery, improving the success rate of surgery and enhancing patients’ prognosis.

Establishment of a PCR-RFLP method for detecting mutations in genes related to insecticide resistance in Aedes albopictus and Culex tritaeniorhynchus

LIU Juan, LIU Peng, WANG Yawei, YU Xiaomei, QIU Xinghui

2024, 42(6):  744-747.  doi:10.12140/j.issn.1000-7423.2024.06.008

Asbtract ( )   HTML ( )   PDF (1494KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To establish a rapid detection method for detecting mutations in genes related to insecticide resistance in Aedes albopictus and Culex tritaeniorhynchus based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology. Methods PCR primers were designed based on the gene sequences of voltage-gated sodium channel (VGSC) in Ae. albopictus, VGSC and acetylcholinesterase (AChE) in Cx. tritaeniorhynchus, and the restriction enzyme cutting sites were optimized to establish a PCR-RFLP method for detecting mutations in genes related to insecticide resistance in Ae. albopictus and Cx. tritaeniorhynchus. Mosquitoes were collected by light trap, and genomic DNA was extracted from individual mosquitoes. PCR was performed using genomic DNA from individual mosquitoes as templates to amplify fragments containing the 1016 site of the Ae. albopictus VGSC gene, the 1014 site of the Cx. tritaeniorhynchus VGSC gene and the 455 site of the Cx. tritaeniorhynchus AChE gene. The PCR products of 3 genes were digested with the restriction enzymes BsaJ Ⅰ, Dde Ⅰ and Mbo Ⅰ, respectively, and the genotypes of individuals were distinguished based on the length polymorphism of the digested fragments. Results The detection method for the VGSC-V1016G mutation in Ae. albopictus, the VGSC-L1014F mutation and the AChE-F455W mutation in Cx. tritaeniorhynchus was established successfully. The enzyme digestion results showed that among the five Ae. albopictus VGSC amplification products, three were homozygous for the wild-type 1016V, and two were heterozygous for the wild-type 1016V and mutant 1016G. Among the five Cx. tritaeniorhynchus VGSC amplification products, two were homozygous for the wild-type 1014L, two were heterozygous for the wild-type 1014L and mutant 1014F, and one was homozygous for the mutant 1014F. Among the five Cx. tritaeniorhynchus AChE amplification products, two were heterozygous for the wild-type 455F and mutant 455W, and three were homozygous for the mutant 455W. Conclusions The PCR-RFLP method established for detecting mutations in genes related to insecticide resistance in Ae. albopictus and Cx. tritaeniorhynchus is convenient, cost-effective and highly accurate.

Establishment and assessment of nucleic acid detection method for Ancylostoma duodenale based on RPA-CRISPR/Cas12a technology

YAN Shuning, YANG Hanyin, CAI Yuchun, XU Bin, YU Chenghang, MO Ziran, LU Yan, YANG Shuo, XIN Yi, ZHENG Bin

2024, 42(6):  748-755.  doi:10.12140/j.issn.1000-7423.2024.06.009

Asbtract ( )   HTML ( )   PDF (3432KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To develop a rapid, convenient, and efficient nucleic acid method based on RPA-CRISPR/Cas12a technology for detecting Ancylostoma duodenale in human stool samples to facilitate timely intervention, reduce the risk of disease transmission, and enhance the level of public health. Methods Based on the conserved sequence of mitochondrial cytochrome c oxidase subunit Ⅰ (cox1) of A. duodenale, RPA primers and CRISPR RNA (crRNAs) were designed and screened, while the concentration of crRNA and ssDNA, along with other reaction conditions, were optimized to establish the RPA-CRISPR/Cas12a fluorescence detection system. The minimum detection limits were determined using A. duodenale plasmid samples as templates, at the concentrations of 10⁵, 10⁴, 10³, 10², 10¹, 10⁰, 10⁻¹ and 10⁻² copies/μl, employing both the RPA method and the RPA-CRISPR/Cas12a method. The specificity of the RPA-CRISPR/Cas12a method was determined using genomic DNA templates from A. duodenale (10⁵ copies/μl), Necator americanus (20 ng/µl), Anisakis (30 ng/µl), and Trichinella spiralis (60 ng/µl). To assess the sensitivity and specificity of the RPA-CRISPR/Cas12a method, Kato-Katz combined with semi-nested PCR were used as reference standards, and 42 fecal samples (31 positive and 11 negative for A. duodenale based on the Kato-Katz method) were tested. Sensitivity and specificity values for the RPA-CRISPR/Cas12a method were calculated, and the agreement between detection results was analyzed using Kappa statistics. Results A RPA-CRISPR/Cas12a fluorescence detection system targeting the A. duodenale cox1 gene sequence was developed. In the optimized CRISPR/Cas12a system, the final concentrations of LbCas12a nuclease and crRNA probe were both 50 nmol/L, while the ssDNA reporter concentration was 100 nmol/L. The reaction was performed at 37 ℃ for 40 minutes. The minimum detection limit for the RPA assay was 10⁰ copies/μl, while the RPA-CRISPR/Cas12a system achieved a lower detection limit of 10 copies/μl, demonstrating enhanced sensitivity. The system showed no cross-reactivity with N. americanus, Anisakis, and T. spiralis. When compared with the Kato-Katz combined with semi-nested PCR, the RPA-CRISPR/Cas12a assay exhibited a sensitivity of 100% (19/19), a specificity of 91.30% (21/23), and good consistency (Kappa = 0.9048). Conclusion The RPA-CRISPR/Cas12a fluorescent detection system established in this study possesses rapid, sensitive, and specific detectability, providing reliable technical support for public health surveillance and the prevention and control of hookworm disease.

Analysis of factors and warning indicators for the intensification of imported malaria in Changsha

ZHOU Quan, WEI Chaoxia, CAI Chunlin, LI Jinqiang

2024, 42(6):  756-762.  doi:10.12140/j.issn.1000-7423.2024.06.010

Asbtract ( )   HTML ( )   PDF (1469KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the characteristics of imported malaria in Changsha and explore the influencing factors and early warning indicators of severe malaria patients in combination with laboratory indicators. Methods A retrospective analysis was conducted on the case data of malaria patients hospitalized in the First Hospital of Changsha from 2012 to 2023. Differences between non-severe and severe malaria patients were compared. Bivariate correlation-Spearman correlation coefficient and logistic regression analysis were used to observe the correlation and influence degree between indicators and severe malaria. Indicators strongly related to prognosis were screened to identify independent impact factors for the progression of malaria patients to severe cases. Receiver operating characteristic (ROC) curve analysis was performed to assess the efficacy of individual and combined indicators in early prediction of severe malaria, and the area under the curve (AUC) was calculated. Results Case data from 127 malaria patients were collected, which including 38 severe malaria patients and 89 non-severe patients. Compared with non-severe malaria patients, severe malaria patients had longer durations from onset to hospitalization [4.00 (2.75, 6.00) d] and longer hospitalization time [7.00 (4.75, 12.00) d] (Z = -2.20, -3.05, P < 0.05, P < 0.01). More severe malaria patients [78.9% (30/38)] were presenting with occult blood in urine (χ² = 18.67, P < 0.01). They had lower red blood cell count [4.11 (3.46, 4.65) × 10¹²/L], platelet count [(53.50 ± 53.73) × 10⁹/L] and albumin level [(34.99 ± 5.36) g/L] (Z = -2.37, t = 6.72, t = 4.10; P < 0.05, P < 0.01, P < 0.01), but higher levels of alanine aminotransferase (ALT) [53.50 (38.63, 73.75) U/L], aspartate aminotransferase (AST) [50.85 (34.00, 97.05) U/L], lactate dehydrogenase (LDH) [449.50 (321.50, 625.75) U/L], total bilirubin [(74.18 ± 78.09) μmol/L] and procalcitonin (PCT) [2.95 (0.91, 19.47) ng/ml] (Z = -3.51, Z = -3.48, Z = -3.58, t = -3.91, Z = -4.63; all P < 0.01) than non-severe patients. Durations from onset to hospitalization, hospitalization time, urinary occult blood and levels of ALT, AST, total bilirubin, LDH, and PCT were positively correlated with severe malaria (r > 0, all P < 0.05), while red blood cell count, platelet count and albumin level were negatively correlated with severe malaria (r < 0, all P < 0.05). Among them, platelet count was a good indicator of malaria severity (|r| ≥ 0.5, P < 0.01). Urinary occult blood, AST, total bilirubin, LDH, serum creatinine, urea nitrogen and PCT were risk factors for severe malaria (OR > 1, P < 0.05), while red blood cell count, platelet count, hemoglobin and albumin level were protective factors against severe malaria (OR < 1, P < 0.05). Platelet count and total bilirubin level were independent impact factors for the progression of malaria patients to severe cases (P < 0.05), with AUCs of 0.867 and 0.769, respectively. The AUC for the combined detection of the both was 0.900, which was higher than that for single indicator detection (P < 0.01). Conclusion Platelet count and total bilirubin level can serve as independent impact factors for early warning of severe malaria. Combined detection is more helpful than single detection for early warning of severe malaria.

STANDARD INTERPRETATION

Interpretation of criteria for Detection of Leishmania: Smear Microscopy Exanimation

WEI Furong, YU Chenghang, ZHENG Bin, GAO Chunhua

2024, 42(6):  763-766.  doi:10.12140/j.issn.1000-7423.2024.06.011

Asbtract ( )   HTML ( )   PDF (1140KB) ( )  

References | Related Articles | Metrics

In accordance with the Management Measures for Health Criteria and GB/T 1.1-2020 Standardization Work Guidelines, Detection of Leishmania - Smear Microscopic Exanimation (WS/T 10018-2024) (referred to as the Criteria) was compiled. The Criteria consists of eight chapters, including “the range of application”, “normative references”, “terms and definitions”, “instruments and equipment”, “reagent or materials”, “detection steps”, “result determination”, and “waste disposal”, attached with four informative appendices (three detection methods, reagent preparation). The Criteria was issued by the National Disease Control and Prevention Administration (National Center for Disease Control Circular [2024] No. 1) and will be formally implemented at start on March 1, 2025. The Criteria provides technical guideline specifications for smear microscopy in detecting Leishmania in medical institutions and disease prevention and control institutions. Combined with the current endemic situation of kala-azar in China, this paper interprets the main content of the Criteria to promote the publicity and implementation.

REVIEWS

Tick-derived anticoagulant proteins and their potential value in the medical field

HE Xiaofei, GAO Jinliang

2024, 42(6):  767-775.  doi:10.12140/j.issn.1000-7423.2024.06.012

Asbtract ( )   HTML ( )   PDF (1241KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Ticks are hematophagous insects that infest both humans and animals. During a blood meal, they inevitably activate the host’s coagulation system. To evade the host’s defense mechanisms, ticks secrete various anticoagulant proteins that regulate the host’s coagulation system, ensuring continuous blood flow. Most anticoagulation proteins are protease inhibitors capable of interacting with coagulation factors or molecules associated with platelet activity to effectively regulate the host’s blood coagulation process. So far, numerous tick-derived anticoagulant molecules with diverse structures have been identified. These molecules exhibit high specificity and strong affinity for their target molecules in the host. Anticoagulants developed from these tick-derived molecules are expected to offer effective anticoagulant activity with a lower risk of bleeding. This article reviews the research progress of some tick-derived anticoagulant molecules, delving into their mechanisms of action and potential clinical application, aiming to inform the development of new anticoagulant drugs.

Research progress on diagnostic techniques for sparganosis mansoni

GUO Jiatian, LIN Lin, LU Yan

2024, 42(6):  776-782.  doi:10.12140/j.issn.1000-7423.2024.06.013

Asbtract ( )   HTML ( )   PDF (617KB) ( )  

References | Related Articles | Metrics

Sparganosis mansoni is a zoonotic parasitic disease caused by infection with plerocercoid, the larval stage of Sparganum mansoni, which is prevalent globally and poses a great threat to human health. S. mansoni plerocercoid may migrate in the human body, and misdiagnosis and missed diagnosis of sparganosis mansoni are common because of variable sites of parasitization, diverse clinical symptoms and lack of specific manifestations. Precision diagnosis of sparganosis is therefore of great importance. Currently, a variety of assays have been used for the diagnosis of human sparganosis mansoni. This review summarizes the advances in the pathogenic, imaging, immunological and molecular biological techniques for diagnosis of sparganosis mansoni, so as to provide insights into further studies on the diagnostic techniques of sparganosis mansoni.

Advances in surgical treatment of hepatic alveolar echinococcosis

ZULIPIKAER Tusunniyazi, JIANG Tiemin, WEN Hao

2024, 42(6):  783-789.  doi:10.12140/j.issn.1000-7423.2024.06.014

Asbtract ( )   HTML ( )   PDF (1251KB) ( )  

References | Related Articles | Metrics

Hepatic alveolar echinococcosis (HAE) is a zoonotic disease. It is predominantly found in regions such as Xinjiang, Qinghai, Xizang and Sichuan in China. HAE can cause severe and irreversible damage, and significantly impacting human health and regional economies. The prevention and control of HAE remains a key focus of national disease prevention and control efforts. Radical surgical resection is the preferred treatment for HAE; while palliative surgery and liver transplantation are alternative options for late-stage HAE cases radical resection is not suitable. This article provides an overview of the current treatment status and latest advancements in the management of HAE.

Research progress of macrophage regulation of liver fibrosis induced by alveolar echinococcosis

YIN Xianmin, CHONG Shigui, CHEN Gen, QIN Junmei, ZHAO Yumin

2024, 42(6):  790-795.  doi:10.12140/j.issn.1000-7423.2024.06.015

Asbtract ( )   HTML ( )   PDF (1662KB) ( )  

Figures and Tables | References | Related Articles | Metrics

Alveolar echinococcosis (AE) is a zoonotic parasitic disease that poses a serious threat to human health and the development of animal husbandry. The augmented infiltration of various immune cells (including macrophages) around AE lesions can lead to fibrosis in various important organs, primarily the liver, and might progress into cirrhosis, liver cancer, and other serious disorders. Several types of macrophages manifest as M1 or M2 macrophages at different stages of AE liver inflammation and fibrosis formation, secreting pro-inflammatory and anti-inflammatory cytokines to promote the clearance of echinococcus in the early stage and exerting an anti-inflammatory effect in the middle and late stages, thereby playing a regulatory role in AE liver fibrosis formation. In this article, the regulatory mechanisms of several types of macrophages in AE-induced liver fibrosis are discussed.

SHORT COMMUNICATIONS

Molecular detection and genetic characteristics of Cryptosporidium spp. in Equus animals in some areas of Anhui Province

WANG Jipu, ZHANG Yujin, WU Xinghua, WANG Mingxing, ZHU Yuqiao, LUO Hui, GAO Sichao, LIU Xinchao, LI Wenchao

2024, 42(6):  796-801.  doi:10.12140/j.issn.1000-7423.2024.06.016

Asbtract ( )   HTML ( )   PDF (1501KB) ( )  

Figures and Tables | References | Related Articles | Metrics

To investigate the infection status and genetic characteristics of Cryptosporidium in Equus animals in Anhui Province, fresh fecal samples were collected from three horse farms in Chuzhou, Suzhou and Anqing, as well as one donkey farm in Fuyang between November 2019 and January 2022. DNA was extracted from the fecal samples, and nested PCR was used to amplify the Cryptosporidium small subunit ribosomal RNA (SSU rRNA). For samples that tested positive for SSU rRNA, nested PCR was further performed to amplify the 60-kDa glycoprotein (gp60) gene. Sequencing of the nested PCR positive products was conducted, followed by sequence alignment to analyze the Cryptosporidium species/genotypes/subgenotypes. A total of 455 fecal samples from Equus animals were collected, with 7 samples positive for the Cryptosporidium SSU rRNA, resulting in an infection rate of 1.5% (7/455). The infection rates for horses and donkeys were 1.9% (3/159) and 1.3% (4/296), respectively. The 3 horse-derived SSU rRNA sequences showed 100% identity with the sequences of Cryptosporidium horse genotype (MK775040, ON384432), and the gp60 sequences exhibited 100% identity with the sequence of ⅥaA15G4 subtype (MK761060). The 4 donkey-derived SSU rRNA sequences had 99.58% identity with the C. hominis (KR296813) sequence, and the gp60 sequences showed 100% identity with the ⅠkA16G1 subtype (KU200963, KU200958 and KU200962) sequences. The results of this study indicate that Cryptosporidium infections exist in Equus animals in Anhui Province, with the main species being the Cryptosporidium horse genotype and C. hominis.

Establishment and application of a recombinant polymerase amplification method for detecting Otodectes cynotis

SONG Shaozheng, MENG Yaqin, WU Yingchao, YU Kangying, ZHOU Zhe

2024, 42(6):  802-805.  doi:10.12140/j.issn.1000-7423.2024.06.017

Asbtract ( )   HTML ( )   PDF (718KB) ( )  

Figures and Tables | References | Related Articles | Metrics

To establish a recombinant polymerase amplification (RPA) method for detecting Otodectes cynotis, this study used the conserved 18S-ITS1 gene sequence of O. cynotis as the target sequence, designed and synthesized RPA amplification primers and probes, established an RPA reaction system, and evaluated the sensitivity and specificity of the method. After identifying 150 clinical samples of suspected O. cynotis infection in canine earwax using fluorescence quantitative PCR, the RPA detection method and microscopic examination method were used to detect the difference in positive detection rate between the two methods. The results showed that the RPA reaction system constructed consisted of 5 mg recombinant polymerase, 10 μl reaction buffer, 2 μl OC18-F primers, 2 μl OC18-R primers, 1 μl labelled fluorescent probe, 1 μl DNA template, 0.5 μl 500 mmol/L magnesium acetate, and sterilized ultrapure water added to make up to 50 μl. The amplification conditions were 39 ℃ for 30 minutes, and the sensitivity of detecting O. cynotis was 1 copy/μl, with high specificity. There was no cross-reactivity with other mite species such as O. cati, Psoroptes cuniculi, Sarcoptes scabiei, Demodex brevis, D. folliculorum and P. natalensis. The positive detection rate of RPA on clinical samples of canine earwax suspected of infection with O. cynotis was 71.33% (107/150), which was higher than the detected by microscopy with 42.67% (64/150) (χ² = 10.813, P < 0.05). The positive consistency rate between RPA detection and fluorescence quantitative PCR detection is 100%. The RPA method established in this study for detecting O. cynotis has the characteristics of fast detection speed, high sensitivity, strong specificity and high accuracy, which provid reference for the clinical diagnosis of O. cynotis disease.

Investigation on the Toxoplasma gondii serum antibodies among patients with malignant tumors in Dali, 2020-2022

DU Shutong, YANG Yiwei, WU Tianfei

2024, 42(6):  806-809.  doi:10.12140/j.issn.1000-7423.2024.06.018

Asbtract ( )   HTML ( )   PDF (1336KB) ( )  

Figures and Tables | References | Related Articles | Metrics

The study included 721 patients with 12 types of malignant tumors treated at the First Affiliated Hospital of Dali University from November 2020 to May 2022 and a control group of 100 healthy individuals undergoing physical examinations during the same period. Basic information and serum samples were collected, and ELISA was used to detect the presence of anti-Toxoplasma gondii IgG and IgM antibodies in the serum samples. Statistical analysis was performed using SPSS 26.0 software. The results showed that among the 721 patients with malignant tumors, the breakdown by cancer type included lung cancer (94 cases), rectal cancer (90 cases), breast cancer (92 cases), colon cancer (69 cases), stomach cancer (74 cases), liver cancer (63 cases), thyroid cancer (60 cases), bladder cancer (40 cases), cervical cancer (30 cases), endometrial cancer (31 cases), esophageal cancer (43 cases), and prostate cancer (35 cases). There were 418 male cases and 403 female cases, with an average age of (57.27 ± 12.23) years. A total of 361 cases were of the Han ethnic group and 460 cases were of the minority group. Three hundred and twenty-four cases were from urban areas, while 497 cases were from rural areas. The positivity rate of anti-T. gondii antibodies in malignant tumor patients (32.04%, 231/721) was significantly higher than that in the control group (13.00%, 13/100) (χ² = 14.14, P ˂ 0.01). The positive rate of anti-T. gondii IgG antibodies (21.22%, 153/721) were higher than that of healthy controls (8.00%, 8/100) (χ² = 9.74, P < 0.05). There was no statistically significant difference in the positive rate of anti-T. gondii IgM antibodies (10.82%, 78/721) compared to healthy controls (5.00%, 5/100) (χ² = 2.57, P > 0.05). The positive rates of anti-T. gondii IgG antibodies in patients with lung cancer, rectal cancer, breast cancer, colon cancer, stomach cancer, liver cancer, thyroid cancer, bladder cancer, and cervical cancer were 21.28% (20/94), 22.22% (20/90), 17.39% (16/92), 26.09% (18/69), 25.68% (19/74), 19.05% (12/63), 21.67% (13/60), 25.00% (10/40), 26.67% (8/30), respectively, which were higher than those in the healthy control group of 8.00% (8/100) (χ² = 6.92, 7.63, 3.86, 10.26, 10.14, 4.38, 6.14, 7.37, 5.82; P ˂ 0.05). There was no significant difference in the positive rate of anti-T. gondii IgG antibody between other cancer types and the control group (P > 0.05). The positive rate of anti-T. gondii IgM antibodies were the highest in patients with breast cancer at 18.48% (17/92), which was significantly higher than that in the control group of 5.00% (5/100) (χ² = 8.58, P ˂ 0.05). There was no significant difference in the positive rate of anti-T. gondii IgM antibody in other types of cancer compared with the healthy control group (P > 0.05). Among 12 different types of malignant tumor patients, cervical cancer patients had the highest anti-T. gondii IgG positive rate at 26.67% (8/30). The positive rate of anti-T. gondii IgM antibody in breast cancer patients was 18.48% (17/92). However, there was no statistically significant difference in the positivity rates of anti-T. gondii IgG and IgM antibodies among different cancer (χ² = 3.96, 11.26, P > 0.05). The positive rate of T. gondii serum in patients with malignant tumors in Dali is higher than that in healthy individuals. It is recommended that T. gondii testing be conducted alongside cancer treatment.

Genetic identification on Meotrchis orientalis infection in human populations of Pucheng County, Fujian Province

CHEN Yunhong, XIE Xianliang, CHEN Baojian, CHEN Fangmei, YU Wenwu

2024, 42(6):  810-812.  doi:10.12140/j.issn.1000-7423.2024.06.019

Asbtract ( )   HTML ( )   PDF (1390KB) ( )  

Figures and Tables | References | Related Articles | Metrics

To understand the infection status of Meotrchis orientalis among the population in Pucheng County, Fujian Province and supplement the biological information of M. orientalis. Fecal samples of individuals infected with M. orientalis reported from the national clonorchiasis surveillance site of Pucheng County from 2016 to 2020 were collected. Modified Kato-Katz thick smear method (two slides per sample) was used to examine the eggs of M. orientalis. The DNA was extracted from fecal samples. The internal transcribed spacer (ITS) sequence of M. orientalis ribosomal DNA was amplified by PCR and sequenced. The positive rates of 2 methods were calculated and compared by Chi-square test. A total of 13 fecal samples of individuals infected with M. orientalis were collected. Among them, 11 tested positive using Kato-Katz method; Target bands at 262 bp were amplified using PCR method in 10 fecal samples, which showed a 99.6% identity to the ITS sequence of M. orientalis (MK482055). The positive rates of Kato-Katz method and PCR method were 11/13 and 10/13, respectively, with no statistically significant difference (χ² = 0.25, P > 0.05). The results indicated that there was an infection of M. orientalis among the population in Pucheng County, which was easily confused with Clonorchis sinensis infection. The ITS sequence used in this study can distinguish between M. orientalis and C. sinensis, and can be used as a genetic marker for M. orientalis.

CASE REPORTS

A child case of complicated Paragonimus infection

CAI Zezheng, YANG Ping, LIAO Qianqian

2024, 42(6):  813-816.  doi:10.12140/j.issn.1000-7423.2024.06.020

Asbtract ( )   HTML ( )   PDF (1977KB) ( )  

Figures and Tables | References | Related Articles | Metrics

A 13-year-old boy, a student from Jianhe County, Guizhou Province, was admitted to the People’s Hospital of Qiandongnan Miao and Dong Autonomous Prefecture on September 15, 2023, due to “repeated coughing for more than 20 days”. On August 26, 2023, he was diagnosed with "pleural effusion and community-acquired pneumonia" at an external hospital and received pleural effusion drainage and medication, which improved his condition. The chest CT scan showed a small amount of pleural effusion. Physical examination revealed a subcutaneous mass around the navel with skin itching. Laboratory tests revealed an elevated eosinophil count (2.28 × 10⁹/L), eosinophil percentage (20.1%), and serum total IgE antibodies (1 101.77 IU/ml). IgG antibodies against Paragonimus, Clonorchis sinensis, and Sparganum were all positive. The epidemiological investigation results showed that the child had a history of eating snake meat and drinking unboiled water. Therefore, a diagnosis of parasitic infection with a high possibility of Paragonimus infection was made. After two courses of oral praziquantel treatment (1.2 g, three times daily, for 5 days and then 3 days), the child’s cough, subcutaneous mass around the navel, and skin itching were significantly relieved, and the eosinophil count returned to normal. He was discharged on September 26, 2023. After 4 months, a follow-up showed slight encapsulated pleural effusion with mild pleural thickening. The pleural effusion resolved after 10 months of follow-up.

A case report of hepatic cystic echinococcosis with a latent infection period of 25 years

SUN Li, LU Enchang, CHENG Meng, AI Yanhong

2024, 42(6):  817-819.  doi:10.12140/j.issn.1000-7423.2024.06.021

Asbtract ( )   HTML ( )   PDF (1286KB) ( )  

Figures and Tables | References | Related Articles | Metrics

A 45-year-old male patient, a driver from Xiangyang, Hubei, presented to Xiangyang Hospital of Traditional Chinese Medicine on July 9, 2023, with a “liver mass detected for more than 4 days”. Upper abdominal MRI revealed a huge cystic lesion in the right lobe of the liver, with T1 long/T2 slightly long signal intensity on MRI. The DWI shows a slightly hyperintense signal, with dimensions of approximately 139 mm × 119 mm × 147 mm. Multiple small cysts were visible within the lesion, arranged in a wheel-like pattern around the periphery. Immunoserological testing of the patient’s venous blood showed positive IgG antibodies against Echinococcus. On July 15, a partial hepatectomy was performed. The resected mass was cystic, grayish-white and measured 160 mm × 150 mm × 150 mm in size. The contents of the mass were a pale yellow, clear liquid, with varying-sized cystic cavities inside. Pathological examination of the mass revealed a cyst wall thickness of approximately 1 mm with multiple protoscoleces present. The patient had served in the military in Xizang from 1996 to 1999, during which time he had a history of contact with cattle and sheep feces, as well as a history of drinking unboiled water or consuming food in the wild. Based on the epidemiological investigation, clinical data, and relevant examination results, the patient was ultimately diagnosed with imported hepatic alveolar echinococcosis from Xizang, with an incubation period of up to 25 years. The patient recovered well after surgery and was discharged one week later. Upon discharge, the patient was instructed to take albendazole orally (15 mg/kg per day). Two weeks after surgery, the patient reported no significant discomfort.

A case of visceral leishmaniasis with Aspergillus niger infection in Shaanxi Province

XING Jia, YU Yanli, WANG Jianjun, XU Shijuan

2024, 42(6):  820-822.  doi:10.12140/j.issn.1000-7423.2024.06.022

Asbtract ( )   HTML ( )   PDF (1097KB) ( )  

Figures and Tables | References | Related Articles | Metrics

A woman aged 55 years was admitted to Department of Hematology, Shaanxi Nuclear Industry 215 Hospital on April 17, 2024 because of abnormal blood cell parameters for more than 9 months. Multiple routine blood tests showed progressive reductions of blood cells, with the lowest white blood cell count of 1.4 × 10⁹/L, hemoglobin levels of 90 g/L, and platelet levels of 90 × 10⁹/L. The patient was given intermittently cell-boosting treatments with Diyushengbai tablets and granulocyte colony stimulating factors, resulting in difficulty in maintaining white blood cell counts. The case had worked outdoors for a long period of time, and complained of mosquito and sandfly bites. Admission physical examinations showed a body temperature of 37 ℃, blood pressure of 73/43 mmHg (1 mmHg = 0.133 kPa), poor mental health, facial appearance of chronic diseases and soybean-sized scabs and scales scattered on the skin. CT shows slightly enlarged liver, enlarged spleen, widened portal vein, and enlarged spleen. Bone marrow aspiration revealed delayed maturation of megakaryocytes, hypochromic anemia of small cells, and detection of Leishmania-Donovan body in macrophages. Bone marrow biopsy showed phagocytosis of phagocytic cells containing parasites or microorganisms. Metagenomics capture (MetaCAP) pathogen detection showed that the sequence number of L. infantis was 371 554, and the sequence number of Aspergillus niger was 86 880. The diagnosis was visceral leishmaniasis with A. niger infection. Following administer sodium antimony gluconate (6 ml/d) in combination with light shielded pump administration of amphotericin B cholesterol sulfate complex (50 mg/d) for treatment for two weeks, the case had improved symptoms and was discharged from hospital. One-month follow-up showed that routine blood tests returned to normal, and no other remarkable abnormalities.

A case of eosinophilia secondary to cutaneous fly maggot disease

DONG Cong, YIN Qichao, LI Yuhong

2024, 42(6):  823-826.  doi:10.12140/j.issn.1000-7423.2024.06.023

Asbtract ( )   HTML ( )   PDF (3580KB) ( )  

Figures and Tables | References | Related Articles | Metrics

A 43-year-old female patient from Yushu, Qinghai, presented to the respiratory department of the Affiliated Hospital of Qinghai University due to “intermittent chest and back pain discomfort for more than 4 months” In September 2022. Physical examination upon admission revealed subcutaneous nodules approximately 1 cm × 0.5 cm in size, palpable on the anterior chest, above the right upper limb elbow joint, and on the left lower limb knee joint, with significant tenderness. Blood test showed an eosinophil count of 15.58 × 10⁹/L, an eosinophil percentage of 67.4% and an erythrocyte sedimentation rate of 17.00 mm/h. Bone marrow examination results indicated active hematopoietic tissue proliferation in the bone marrow, with a significant increase in eosinophils, accounting for approximately 62% of nucleated cells. Skin biopsy pathology revealed focal lymphocyte, plasma cell, and eosinophil infiltration around blood vessels and between adipose lobules. Treatment with oral prednisone acetate tablets (60 mg/d) for one month did not alleviate the pain in the subcutaneous nodules. In November 2022, a total of 5 fly larvae emerged successively from subcutaneous sites on the patient’s left ear, right lower limb, left upper limb, neck and anterior chest. Morphological identification confirmed them as Hypoderma bovis larvae. The patient is a herdsman residing in a plateau pastoral area for a long time, with a history of sleeping on grass and drying clothes on the grass during summer. Based on the epidemiological investigation, clinical manifestations, and relevant examination results, the final diagnosis was secondary eosinophilia due to cutaneous myiasis caused by H. bovis larvae. After confirmation of the diagnosis, the patient was treated with oral albendazole (0.4 g/d) and loratadine tablets (10 mg/d) for 13 days. A follow-up blood test showed that the eosinophil count returned to normal, and the patient’s symptoms improved. During a one-year telephone follow-up, the patient did not report any recurrence of skin pain or other discomfort.