Objective To understand the situation of soil-transmitted nematode infection in China in 2020 and provide support for evaluating the development of surveillance on soil-transmitted nematodiasis in various provinces/municipalities/autonomous regions, and improving and perfecting the control strategies. Methods Surveillance was carried out in 408 national surveillance sites (counties) in 31 provinces (municipalities, autonomous regions) in China in 2020. With the county as unit, each site was divided into 5 areas geographically: east, west, south, north, and central part, followed by selecting one township (town), and therein one administrative village was selected from wherein, 200 permanent residents over the age of 3 were sampled. A total of 1 000 people were surveyed at each surveillance site. Fecal samples were collected from the sampled villagers, and examined by using the modified Kato-Katz thick smear method (two slide-reading for each sample) for infection of hookworm, Ascaris lumbricoides, Trichuris trichiura and Enterobius vermicularis, to calculate the infection rate and intensity, respectively. In addition, soil samples were collected from fields or vegetable gardens of each village in the survey site, and examined for hookworm larvae using 5% saline at 45 ℃, and for Ascaris eggs by saturated sodium nitrate flotation method. Results In 2020, the overall infection rate of soil-transmitted nematode in residents was 0.84% (3 485/415 672) in 408 surveillance sites of 31 provinces (municipalities/autonomous regions), with the highest found in Hainan (6.34%, 199/3 141), followed by Yunnan (5.80%, 963/16 616) and Sichuan (3.66%, 592/16 168); infection rate in females was 0.91% (1 944/213 591), which was higher than that of 0.76% in males (1 541/202 081) (χ² = 27.20, P < 0.01). The soil-transmitted nematode infection rate was the highest in the age group ≥ 60-years-old, which is 1.26% (1 376/109 251). The difference between each age group was statistically significant (χ² = 382.28, P < 0.01). The infection rates of hookworm, A. lumbricoides, T. trichiura were 0.51% (2 016/415 672), 0.19% (805/415 672) and 0.16% (673/415 672), respectively. Among them, hookworm and T. trichiura had only mildly infected cases. The proportions of mild and moderate A. lumbricoides infections were 99.25% (799/805) and 0.75% (6/805), respectively. In 2020, 2 604 soil samples were examined and found that the positive rate of Ascaris eggs and hookworms was 3.07% (80/2 604) and 2.42% (63/2 604), respectively. Conclusion In 2020, the infection rate of soil-transmitted nematode in China remains at a low level in general, but the regional differences are still significant, and the areas with high infection rates still exist. At the same time, it is necessary to strengthen the control measures for the key groups of people over age of 60, women and children, and carry out health education.

Objective To analyze the epidemiological characteristics of visceral leishmaniasis cases in Longnan City, Gansu Province from 2005 to 2021, to provide scientific basis for formulating prevention and control strategy for visceral leishmaniasis. Methods The information on the visceral leishmaniasis cases residing at the current address in Longnan City, reported by the National Infectious Disease Surveillance Reporting and Management System from 2005 to 2021, was collected. A database was established using Microsoft Excel 2013. Three-dimentional distribution characteristics of visceral leishmaniasis cases were analyzed with descriptive epidemiological method. The distribution map of reported cases was plotted by Arc GIS 10.2. Statistical analysis was performed using SPSS 25.0 software. Results From 2005 to 2021, a total of 1 109 cases of visceral leishmaniasis were reported in Longnan City,among them, 577 cases were of clinical diagnosis, and 532 confirmed cases. The annual average incidence rate was reported at 2.49/100 000, showing a low prevalence status. The incidence rate reached 2.76/100 000 in 2005, rose to 4.53/100 000 in 2009, and gradually dropped to 0.46/100 000 in 2021, indicating a trend of first increase and then decrease (χ² = 267.561, P < 0.01). The visceral leishmaniasis cases were mainly distributed in Wudu District (606 cases), Wenxian County (323 cases) and Tanchang County (148 cases), accounting for 97.11% of reported cases (1 077/1 109). In some villages/towns of endemic counties, a cluster of high incidence was found. There were 633 male cases and 476 female cases, and the sex ratio was 1.33∶1; the incidence rates in males and females were 2.73/100 000 and 2.23/100 000, respectively (χ² = 2.699, P > 0.05). The number of reported cases in the 0-4 age group accounted for 50.50% (560/1 109) of the total number of reported cases, with the highest incidence rate of 19.62/100 000. The second is the 5-9 age group, with the number of reported cases accounting for 12.80% (142/1 109) of the total number of reported cases, and the incidence rate is 4.84/100 000. It indicated that the incidence rate tends to decrease with age (χ² = 14.942, P < 0.01). The first incidence rate was occupied in children of scattered living, accounting for 41.12% (456/1 109), and the second in peasants, accounting for 24.71% (274/1 109). Conclusion In Longnan City of Gansu Province, visceral leishmaniasis remains at a low prevalent status. The reported cases are mainly of children. There is a recurgence trend in historically endemic counties.

Zoonotic diseases pose a serious threat to human health and ecological security. Climate change facilitates the crossover and spread of zoonotic diseases by affecting pathogens, hosts, vectors, and human activities, therefore threatening global public health. This article summarizes the impact of climate change on the spread of zoonotic diseases and explores the effective strategies based on the One Health approach to protect human health and safety.

Toxoplasma gondii is an obligate intracellular parasite that widely distributes in the world and causes zoonotic toxoplasmosis. In recent years, parasite infection in the brain has been paid more and more attention. T. gondii can cause central nervous system damage, often manifested as depression, schizophrenia, epilepsy and other symptoms. In this paper, the process and mechanism of T. gondii establishing chronic infection through blood-brain barrier, causing central nervous system injury and disease are reviewed.

A 57-year-old female patient from Lucheng District was admitted to Wenzhou Central Hospital on September 28, 2021. The examination on admission showed the chronic disease face and clear consciousness; the skin and mucous membranes had no yellowish staining nor oedema but were scared. The patient was admitted to the cardiology department with “chest tightness pending further investigation” and was later transferred to the chemotherapy department due to cancerous pleural fluid. The patient had a recurrent fever since October 8. Laboratory tests showed the haemoglobin was 86 g/L, the red blood cell count was 2.74 × 10¹²/L suggesting moderate anaemia. The leukocyte count was 12.7 × 10⁹/L, the absolute neutrophil count was 10.6 × 10⁹/L. The patient also had mildly elevated total and indirect bilirubin; urine culture detects mixed growth of more than 3 bacteria, persistent urinary occult blood 2 +. Wright’s stain of the blood smear was performed on October 25, microscopic examination showed that 1-4 ringlets with purplish-red nuclei and blue cytoplasmic could be found in some red blood cells, which was considered to be a parasitic infection. The patient was a long-term resident of urban Wenzhou with no history of domestic or international travel, and no clear history of tick bites. The patient underwent breast-conserving surgery for left breast cancer in 2005, received chemotherapy and targeted drug therapy for multiple lung metastases between 2014 and 2021 and had 12 blood transfusions from June to October 2021. The PCR product of blood DNA amplificated by primers specific to Babesia was sequenced and identified with 99.43% homology to B. microti (GenBank accession: MG674832.1). The diagnosis of Babesia microti infection was confirmed. The patient was treated with oral chloroquine phosphate tablets (0.5 g/d, double the first dose for 5 d) and intravenous clindamycin (600 mg/d + saline 500 ml intravenously for 10 d), with some relief of symptoms, and parasites were still detected on microscopic examination after 5 days. The symptoms of high fever, jaundice and haemolysis caused by Babesia infection somewhat exacerbate the progression of terminal-stage cancer. On November 8, the patient died of multi-organ failure and unsuccessful resuscitation.

Objective To investigate the parasitic infection in domestic dogs in Da Hinggan Ling area, and provide reference for the prevention and control of local zoonotic parasitic diseases. Methods In September 2020, fresh domestic dog feces were collected from dog owner’s household yard of 6 natural villages including Jiabei Village, Baihua Village, Dongshan Village, Tuqiang Town, Xingfu Village and Guyuan Town in the Da Hinggan Ling area, and total DNA was extracted from the fecal samples. PCR amplification was conducted using 13 pairs of parasite universal primers targeting 8 taxonomy groups including Apicomplexa, Amoeba, Diplomonadida, Kinetoplastida, Parabasalia, Nematoda, Platyhelminthes and Microsporidia, and the target gene fragments were obtained after high-throughput sequencing of the amplified products. The product sequences were compared and analyzed in the NCBI database to identify the parasite species, and the detection rate of parasites in domestic dog feces was calculated. Results Parasite DNA were detected in 37 of 202 domestic dog feces, and the total detection rate was 18.31%. A total of 19 species of parasites which belongs to 15 families from 7 phyla were detected, and the detection rate of different species were significantly different (χ² = 69.488, P < 0.05). Thirteen protozoa species and 6 helminth species were detected. The detection rate of Parabodo caudatus in the family Parabodonidae was the highest at 6.44% (13/202). Among the 37 samples in which parasite DNA was detected, the mixed positive ratio was 32.43% (12/37), of which 72.97% (27/37) detected 1 type of parasite, and 2 types, 3 kinds and 4 kinds accounted for 27.03% (10/37), 0.27% (1/37) and 0.27% (1/37) respectively. Jiabei Village had the highest detection rate of parasite DNA in domestic dog feces, which was 37.70% (23/61), followed by Tuqiang Town, with a detection rate of 36.36% (20/55), and Baihua Village, which was not detected (0/13). There was a statistically significant difference in the detection rate of different sampling sites (χ² = 19.717, P < 0.05). Conclusion In Da Hinggan Ling area, domestic dogs were found infected with multiple parasites, showing considerably higher positive rate detected in some parts of the area, thus, there exists risk of infection caninee-source parasites to local residents.

Objective To analyze the genotype and sequence polymorphisms of mitochondrial cytochrome oxidase 1（co1）and NADPH dehydrogenase subunit 1 (nd1) in Echinococcus granulosus from Yunnan Province. Methods Echinococcus of animal origin was collected from cysts isolated from liver or lung lesions of cattle, sheep and free-range pigs at slaughterhouses in Shangri-La, Daguan, Eryuan, Lushui and Weixi counties of Yunnan Province. Echinococcus of human origin was obtained from the focal tissues surgically removed from hospitals in Jianchuan, Yunlong, Longyang and Yulong Counties, then the E. granulosus was selected after pathogenic identification, and used to extract genomic DNA with a DNA extraction kit, followed by PCR amplification and sequencing of co1 and nd1 genes. The sequence homology and single nucleotide polymorphisms were analyzed using BLAST aligment. Phylogenetic trees based on co1 and nd1 were constructed by the neighbor-joining method using MEGA-X software. Results A total of 62 samples of Echinococcus were collected, 36 of which were pathogenically confirmed as E. granulosus. A total of 32 gene co1 sequences were successfully sequenced, of which 15 were type G1 and 17 were type G5, with a length of 824 bp and 807 bp, respectively. After submitting to GenBank database, the login numbers obtained were OP413393-OP413402, OP413498-OP413506 and OP420520. A total of 34 nd1 genes were sequenced successfully, of which 11 were type G1 and 23 were type G5, with lengths of 882 bp and 888 bp, respectively. The entry numbers obtained by submitting the GenBank database were OP471626-OP471638. The results of sequence polymorphism analysis showed that 1.94% (16/824) mutation sites in the co1 type G1 sequence and 3.10% (25/807) mutation sites in the G5 type sequence were found. In the nd1, 0.45% (4/882) and 2.93% (26/888) mutation sites were found in the type G1 sequence and the G5 type sequence, respectively. The results of multiple genotypic sequence comparison showed that there were 6 homologous sequences of the co1 gene, 4 of which were type G1, the genetic distances of homologous sequences were 0.000 0-0.001 1, 0.000 0-0.000 6, and 0.000 0-0.001 7, respectively. Also included two G5 types. The genetic distances of homologous sequences were 0.000 0-0.002 5 and 0.000 0-0.001 2, respectively. There were 3 homologous sequences in nd1 gene, of which 1 was type G1, and the genetic distance was 0.000 0-0.001 1, 2 of which were type G5, genetic distance was 0.000 0 and 0.000 0-0.001 1. Phylogenetic tree analysis showed that the phylogenetic tree based on co1 and nd1 genes formed two branches of type G1 and type G5, type G1 was closely related to the gene sequences of central and western China (Qinghai, Ningxia, Gansu, Sichuan), Bhutan, Iran and Jordan in the Middle East (GenBank entry number AF297617.1, KJ628328.1, EU072106.1, MW138946.1, AB688602.1). The G5 Type was closely related to Vietnam, China's Guangxi, the United Kingdom, Poland, Zambia, France, Pakistan, Brazil, Kenya and Namibia (GenBank login numbers MW558412.1, MN058591.1, KU378107.1, MZ322608.1, KU743915.1, KU743919.1, MN886291.1, KX010903.1, KU743918.1 and KX138068.1). Conclusion The prevalent isolate of E. granulosus in Yunnan Province are of genotype G1 and G5, and both co1 and nd1 gene reveals single nucleotide polymorphisms.

Objective To establish a rolling circle amplification (RCA) method for detection of Cysticercus pisiformis infection in rabbits. Methods The novel-miR1 derived from C. pisiformis in rabbit serum served as the diagnostic target to establish an RCA diagnostic method for cysticercosis pisiformis fection in rabbit. To establish the RCA method, ligation sequence and the locking probe sequence were designed, and five important reaction conditions were optimized, including the ratio of ligation sequence and padlock probe, ligation reaction time, ligase dosage, amplification reaction time, and dNTP dosage. The sensitivity of the RCA method was assessed, the novel-miR1 standard was serially diluted into 9 samples of different concentrations ranging from 1 fmol/L to 100 nmol/L. The sensitivity and specificity of the optimized RCA methods were assayed using the serum miRNA from 20 healthy rabbits and 20 C. pisiformis infected rabbits (Laboratory preserved samples), and analyzed by the receiver operating characteristic curve (ROC curve) method. To evaluate the application effectiveness of the RCA, 20 female rabbits were infected by gavage with 1 000 eggs of T. pisiformis for collection of serum every month post-infection, which were used to prepare miRNA for application in RCA method under optimized condition. Results The agarose gel electrophoresis results showed that the RCA amplification products remained in the sampling wells of agarose gel, forming a bright band. The tests to optimize RCA condition indicated that the concentration of ligation sequence was 2 μmol/L, padlock probe was 1 μmol/L (ratio of ligation sequence and padlock probe was 2∶1), T4 DNA ligase was 350 U, and the efficacy of ligation reaction was found the highest when ligating for 180 min. The optimal amplification reaction system was 0.5 μmol/L dNTP and amplification reacted 240 min in 100 μl amplification reaction system. Lastly, the RCA method limit of detection was proved to be 10 pmol/L. The RCA method detected showed that the average serum miRNA fluorescence intensity of the samples from healthy rabbits and C. pisiformis infected rabbits were 53.298 ± 1.707 and 97.498 ± 5.892, respectively, which was statistically significant (t = 7.206, P < 0.01). ROC curve showed that the RCA method cut-off value was 61.69 and both sensitivity and specificity were 95%, the area under the curve (AUC) is 0.955 0, likelihood ratio was 19.00. Using the RCA method to detect 20 healthy rabbits’ serum miRNA found 19 samples were negative and 1 positive. The results of RCA detection of the C. pisiformis infected rabbits serum miRNA at different times found that 17 were positive, 2 were suspected, 1 were negative 1 month post-infection. One was negative, and 19 were positive 2 mouths post-infection. Three were negative, and 17 were positive 3 months post-infection. Conclusion An RCA method was established for detecting C. pisiformis infection in rabbits. It was demonstrated that novel-miR1 could be detected by RCA in the serum of rabbits within 3 months post-infection of C. pisiformis, showing good application potential.

In order to identify the infection of Toxoplasma gondii in dogs and cats in Beijing, according to the program of animal disease surveillance in Beijing in 2022, 920 dog serum samples and 816 cat serum samples were collected from scatters, animal hospitals and farms in the whole city. Indirect heamagglutination assay were uesd to detecte Toxoplasma gondii antibodies in dog and cat serum samples. The result shows that the T. gondii antibody positive rate was 5.2% (48/920) in dogs and 7.5% (61/816) in cats. The positive rate of T. gondii antibody in dogs in autumn was 6.8% (30/440), which was higher than that in spring at 3.8% (18/480) with moderate intensity correlation (χ² = 4.37，P < 0.05，0.4 < OR < 0.6). And the positive rate of T. gondii antibody in cats in autumn was 12.5% (50/401), which was higher than that in spring at 2.7% (11/415) with strongly correlated material (χ² = 28.42，P < 0.05，0.1 < OR < 0.3）. In diverse sources, the positive rates of T. gondii antibody in dogs and cats from veterinary clinics were the highest, which were 8.0% (47/590) and 8.5% (61/715) (χ² = 25.31, 9.31，all P < 0.05), respectively. The positive rate of T. gondii antibody in dogs from the six central districts was 11.4%（40/350）. The positive rate of T. gondii antibody in dogs from the outer suburban districts was 1.4% (8/570) with strongly correlated material (χ² = 44.07，P < 0.05，3.0 < OR < 9.9). And the positive rate of T. gondii antibody in cats from the six central districts was 10.3% (37/360). The positive rate of T. gondii antibody in cats from the outer suburban districts was 5.3% (24/456) with moderate intensity correlation (χ² = 7.31，P < 0.05，1.5 < OR < 2.9). This study provided a basis for the further prevention and control of toxoplasmosis in Beijing.

Objective This study investigates the effect of thioredoxin peroxidase (TPx) in the excretory-secretory antigen (ESA) of Cysticercus cellulosae on activation of dendritic cell (DC) in piglets. Methods Healthy piglet medulla-derived DC were cultured in vitroo, in which lipopolysaccharide (LPS) was added at a final concentration of 100 ng/ml on 7 d for stimulation for 2 days, and then continuously cultured for 2 more days to collect immature DC (imDC) and mature DC (mDC) separately. The morphological changes of DCs were observed by light microscopy and scanning electron microscopy on 1 to 9 d of culture. The expression of surface markers CD1 and major histocompatibility complex (MHC-Ⅱ) was detected by flow cytometry. The 7 d imDC was used in the assay with the assigned groups of negative control, TPx, ESA and LPS positive control, to which RPMI 1640 medium, TPx (50 μg/ml), ESA (50 μg/ml) and LPS (100 ng/ml) was added, respectively, to stimulate for 48 h for examining the expression of DC surface markers MHC-Ⅱ, CD80 and CD86 using flow cytometry and for detecting secretion levels of tumor necrosis factor-α (TNF-α), interleukin (IL-6), IL-10, IL-12 in DC culture supernatant by ELISA. One-way ANOVA was used for comparison between multiple groups, and LSD-t test was used for two-way comparison between groups. Results Under ligth microscope, imDC were ovoid in shape with single form at the first day of culture; with the extension of culture time, DC increased in size, appeared pseudopods and spines and other features, and changed from ovoid to irregular shape. Scanning electron microscopy showed that compared with imDC, mDC had irregular morphology, roughly long shuttle shape, and numerous protrusions of different lengths radiating from the cytosol, which were distributed in a dendritic pattern, a typical dendritic structure. Flow cytometry showed that the expression of CD1 and MHC-Ⅱ in imDC was (0.113 ± 0.005)% and (0.430 ± 0.016)%, respectively, which was lower than that of mDC (21.400 ± 0.327)% and (21.333 ± 0.450)% (t = 130.341, 92.906, both P < 0.05). The expression levels of MHC-Ⅱ, CD80, and CD86 in the TPx group were (15.300 ± 0.245)%, (22.900 ± 0.374)% and (13.033 ± 0.249)%, respectively, which were lower than those in the LPS positive control group (19.000 ± 0.374)%, (31.600 ± 0.082)%, and (21.300 ± 0.245)% (t = 11.53, 46.32, 43.84, all P < 0.05) and the ESA group (18.365 ± 0.618)%, (40.400 ± 0.356)% and (30.300 ± 0.283)%] (t = 9.55, 93.17, 91.57, all P < 0.05). The MHC-Ⅱ expression level in the TPx group was higher than that of the negative control group (12.133 ± 0.492)% (t = 9.87, P < 0.05). ELISA results showed that IL-6 level in DC of the TPx group was 15.682 ± 0.660, which was ower than that of 21.041 ± 0.901 in the control group (t = 6.51, P < 0.05); TNF-α (35.711 ± 4.196), IL-6, IL-10 (22.216 ± 1.357) and IL-12 (16.799 ± 0.523) were all lower than those of the LPS positive control group 169.037 ± 7.823, 42.118 ± 1.932, 34.730 ± 1.772, 52.504 ± 2.431 (t = 36.79, 32.09, 13.09, 35.05, all P < 0.05); IL-12 level were lower than that of the ESA group at 23.854 ± 1.020 (t = 6.93, P < 0.05). Conclusion TPx mediates immune tolerance by inducing high expression of DC surface molecules MHC-Ⅱ, low expression of CD80 and CD86, and reducing the secretion levels of IL-6.

Objective To investigate the regulatory effect of galectin-receptor interaction on the small intestine immunopathology of Toxoplasma gondii-infected mice. Methods Eighteen female BALB/c mice were randomly divided into 4 groups: 4 mice in uninfected group (naive group), 4 mice in lactose group (lactose group), 5 mice in T. gondii infection group (Tg group) and 5 mice in T. gondii infection + lactose group (Tg+lactose group). Each mouse in the Tg group and the Tg+lactose group was intraperitoneally (i.p.) injected with 1 000 tachyzoites of T. gondii RH strain, while the naive group and lactose group were i.p. injected with 0.2 ml PBS. Starting from day 0 post infection, each mouse in the Tg+lactose group and the lactose group was i.p. injected with 0.2 ml 0.2 mol/L of lactose, while each mouse in the naive group and the Tg group was i.p. injected with an equal volume of PBS, once in the morning and once in the evening for 7 consecutive days. After infection with T. gondii, the mice survival time in each group was recorded. The mice were euthanized on the 7th day after infection to collect middle segment of jejunum from each mouse for prepareing paraffin sections, which were stained with hematoxylin and eosin (HE) to observe the pathological changes; from the lower segment of jejunum of each mouse, total RNA was extracted and reverse-transcribed, and used in quantitative real-time reverse transcription PCR (qRT-PCR) with β-actin as an internal reference gene to detect the relative mRNA expression level of surface antigen 1 (SAG1), galectin-3, galectin-9, T cell immunoglobulin mucin 3 (Tim-3), leukocyte differentiation antigen 137 (CD137), interleukin 12 (IL-12), interferon-γ (IFN-γ), IL-10, IL-4, transforming growth factor β (TGF-β), chemokine receptor 2 (CCR2) and chitinase 3 like molecule 3 (Ym1). Results After infection with T. gondii, there was no mice died in the naive group and the lactose group. The survival time of the mice in the Tg group was 182-188 h, and the survival time of the mice in the Tg+lactose group was 180-182 h; the difference of the survival time between the two groups was statistically significant (χ² = 19.52, P < 0.05). HE staining showed no inflammation in the mice jejunal tissue in the naive group and lactose group. Shortened intestinal villus, shallower intestinal crypts, necrosis of epithelial cells at the top of villi and inflammatory cell infiltration in the intestinal mucosa were observed in the mice small intestine from the Tg group and Tg+lactose group. Compared with the Tg group, the pathological change of the mice small intestine in the Tg+lactose group was more severe. The qRT-PCR results showed that the relative mRNA expression of SAG1 in the mice small intestine of the Tg+lactose group was 9.17 ± 1.65, which was higher than that in the Tg group (1.00 ± 0.84, t = 4.40, P < 0.05). The relative mRNA expression levels of galectin-3 in the mce small intestine of the naive group, lactose group, Tg group, and Tg+lactose group were 1.00 ± 0.28, 1.71 ± 0.31, 2.46 ± 1.11, and 7.10 ± 1.57, respectively (F = 10.15, P < 0.01). The mRNA expression levels of galectin-9 in the 4 groups were 1.00 ± 0.31, 1.44 ± 0.26, 3.21 ± 1.01, and 7.00 ± 1.08, respectively (F = 14.53, P < 0.01). The mRNA expression levels of Tim-3 in the 4 groups were 1.00 ± 0.12, 0.88 ± 0.28, 1.64 ± 0.31, and 4.89 ± 0.69, respectively (F = 19.15, P < 0.01). The mRNA expression levels of CD137 in the 4 groups were 1.00 ± 0.42, 1.03 ± 0.30, 0.89 ± 0.11, and 3.84 ± 0.77, respectively (F = 8.46, P < 0.01). The mRNA expression levels of IL-12 in the 4 groups were 1.00 ± 0.35, 1.14 ± 0.56, 12.37 ± 4.43, and 18.42 ± 3.89, respectively (F = 10.18, P < 0.01). The mRNA expression levels of IFN-γ in the 4 groups were 1.00 ± 0.56, 1.65 ± 0.53, 5.57 ± 1.84, and 21.26 ± 6.48, respectively (F = 10.38, P < 0.01). The mRNA expression levels of IL-10 in the 4 groups were 1.00 ± 0.20, 1.10 ± 0.25, 8.65 ± 2.52, and 21.98 ± 3.96, respectively (F = 20.84, P < 0.01). The mRNA expression levels of IL-4, TGF-β, and CCR2 in the mice small intestine among the naive group, lactose group, Tg group, and Tg+lactose group had no statistically significant differences (F = 1.09, 4.74, and 2.03, P > 0.05). Ym1 mRNA expression was not detected in the naive group and lactose group, and Ym1 mRNA expression levels between the Tg group and the Tg+lactose group had no statistically significant difference (t = 0.24, P > 0.05). Conclusion Blockage of galectins-receptor interaction in mice infected with T. gondii leads to increased parasite burden in small intesting tissues, and aggravated pathological impairment, as well as upregulated expression of galectin-3, galectin-9, Tim-3, CD137, IL-10 and IFN-γ.

To investigate the molecular identification of Giardia lamblia isolated from an infected case and analyze the source of infection. Fresh fecal samples of the patient and his parents and the nanny were collected for iodine solution staining and microscopic examination. Fecal DNA was extracted, and nested PCR was used to amplify the triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh) and beta-giardin (bg) gene for sequencing. The homology comparison and phylogenetic analysis of gene sequences were performed by BLAST, ChromasPro and MEGA 11.0 software to determine the assemblages. The results showed that there were trophozoites and cysts in the fecal of the patient through microscopic examination. Nested PCR amplified bands of about 500 bp, which were consistent with the tpi, gdh and bg gene fragments of G. lamblia, confirming the infection of G. lamblia. None of the patient’s parents and nanny had diarrhea symptoms, but G. lamblia cysts were found in the stool specimen of his father. Combined with nested PCR and sequencing results, the father was a G. lamblia carrier. The results of the gene comparison showed that the percent identity of tpi, gdh and bg genes sequence amplified from fecal samples of the patient and his father was 99.8%, 100% and 98.5%, respectively. The percent identity of tpi gene sequence amplified from the fecal samples of the patient and his father showed 100% and 99.8% with G. lamblia assemblages AⅡ (GenBank accession no. LC183963), respectively. The amplified gdh gene sequence showed 99.6% and 100% with assemblages AⅡ (GenBank accession no. KF843931), respectively. The percent identity between the bg gene sequence amplified from the patient fecal sample and the sequence of assemblages AⅢ (GenBank accession no. LC183968) is 99.2%, while the percent identity between the bg gene sequence amplified from the father’s fecal sample and the sequence of assemblages AⅡ (GenBank accession no. LC183975) is 100%. The results of phylogenetic tree analysis showed that both the patient and his father were infected with G. lamblia and clustered in the same cluster as assemblages A, and sub-genotype analysis using tpi and gdh as target genes were conducted, which clustered in the same cluster as assemblages AⅡ. Therefore, it can be determined that the patient was infected with G. lamblia assemblages AⅡ, and it could be a family cluster infection caused by internal transmission within the family.

Objective To screen the differentially expressed genes (DEGs) by comparing the transcriptome of the brain tissues between the mice chronically infected with Toxoplasma gondii and normal mice, to analyze the relative transcription level of DEGs in the depression-related kynurenine (KYN) pathway and to provide a theoretical basis for exploring the mechanism of depression-like symptoms caused by Toxoplasma gondii chronic infecttion in mice. Methods SV129 male mice (n = 18) were randomly and equally divided into the infection group and the control group. Mice in the infection group were intraperitoneally injected with 120 tachyzoites of T. gondii ME49 strain (200 μl), and mice in the control group were injected with the same volume of PBS. After 3 months post-infection, mice brain tissues of the two groups were collected for extraction of total RNA to undertake transcriptome sequencing for screening DEG. With the DEGs obtained, cluster analysis, gene ontology (GO) functional annotation analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation and enrichment analysis were performed. Eight DEGs [interferon-γ (IFN-γ), indoleamine 2,3-dioxygenase 1 (IDO1), IDO2, kynurenine-3-monooxygenase (KYNU), kynurenine-3-monooxygenase (KMO), 3-hydroxyanthranilate 3,4-dioxygenase (3-HAO), vimentin (Vim) and brain-derived neurotrophic factor (BDNF)] related to KYN pathway associated with depression were selected to examine each gene’s relative transcription level by quantitative real-time PCR (qRT-PCR), using glyceraldehyde-3-phosphate dehydrogenase gene as an internal reference. Results Transcriptome sequencing found 2 295 DEGs in the brain of the mice from the infection and control groups, of which 2 016 were up-regulated and 279 were down-regulated. GO analysis showed that localisation was the most significantly enriched biological process, with a total of 257 DEGs. The most significantly enriched in cellular components was the protein-containing complex, with a total of 425 DEGs. The most significantly enriched molecular function was molecular transducer activity, with 177 DEGs. The largest number of DEGs enriched in biological process, cell component and molecular function were cell process, cell part and binding, with 1 039, 1 240 and 1 088 DEGs, respectively. KEGG analysis showed that the top three up-regulated metabolic pathways were the immune system, signaling transduction, and viral infectious disease, and the top three down-regulated pathways were signal transduction, signaling molecules and interaction and immune system. Functional enrichment analysis showed that 77 pathways were significantly enriched. The signaling pathways related to depression include tumor necrosis factor signaling pathway, neuroactive ligand-receptor interaction, NF-kappa B signaling pathway, JAK-STAT signaling pathway, necroptosis, apoptosis, chemokine signaling pathway, KYN pathway. The qRT-PCR results showed that the relative transcription levels of IFN-γ, IDO1, IDO2, KYNU, KMO, 3-HAO and Vim genes in the infection group were 3 023.08%, 355.52%, 190.17%, 496.55%, 339.92%, 212.74% and 507.34%, if the relative transcript level of control mice was taken as 100%. Compared with the control group, the transcription was significantly up-regulated (t = 3.782, 3.749, 3.226, 2.908, 2.533, 5.656, 2.948; all P < 0.05 or 0.01). The relative transcription level of BDNF was 63.32%, which was significantly down-regulated (t = 2.398, P < 0.05). The fold change of IFN-γ, IDO1, IDO2, KYNU, KMO, 3-HAO, BDNF, Vim obtained by qRT-PCR was 4.96, 1.74, 0.89, 2.10, 1.60, 1.06, -0.94, 2.18, respectively. The fold change obtained by transcriptome sequencing was 7.30, 0.55, 0.80, 3.83, 2.75, 3.53, -0.86 and 1.93, respectively. The transcriptional trend obtained by qRT-PCR was consistent with that obtained by transcriptome sequencing. Conclusion DEGs from brain tissues of mice chronically infected with T. gondii were screened. Transcriptome analysis revealed that the immune response of central nervous system of the mice with chronic infection of T. gondii was continuously activated. Seven DEGs in KYN pathway related to depression showed up-regulated transcription level.

To understand the malaria epidemiological characteristics in Guizhou Province from 2017 to 2021, the malaria endemic data and individual case information in Guizhou from 2017 to 2021 were collected and analyzed using descriptive statistical methods, species of Plasmodium, source of infection, distribution of cases, clinical visits and diagnosis were analyzed. According to the analysis, 84 malaria cases were reported in Guizhou from 2017 to 2021. Most cases were P. falciparum infection (69.0%, 58/84). All the cases were imported from overseas, 91.7% (77/84) of which were imported from African countries and others were imported from Asia and Oceania. The cases were reported throughout the year, particularly higher in January (15.5%, 13/84). All the cities in Guizhou Province had malaria cases reported, of which Guiyang had the highest number of reported cases (41.7%, 35/84). Of all the reported cases, 92.9% (78/84) were male, and 69.0% (58/84) were aged 30 to 49; 73.8% of the cases were initially diagnosed as malaria. The Center for Disease Control and Prevention (CDC) at the county level had the highest diagnostic accuracy (15/15). The median interval from the onset of symptoms to initial diagnosis was 1 day, and from initial diagnosis to final diagnosis was 3 days. All the cases were reported timely within 1 day after diagnosis, 90.5% (76/84) were undertaken epidemiological investigation within 3 days after reporting, then 98.8% (83/84) of the epidemic sites were surveyed and performed prevention measures within 7 days. This study suggested that to consolidate the achievement of malaria elimination in Guizhou, continuous surveillance of imported malaria cases and regulated disposal measures should be maintained, and the diagnostic and tertiary care capacity should be improved.

After more than 70 years of effective control programme, China has made remarkable achievements in the control of key parasitic diseases, and is moving towards the goal of control and elimination. This paper analyzes the epidemic status and challenges of schistosomiasis, malaria, echinococcosis, leishmaniasis, clonorchiasis and soil-transmitted helminthiasis in recent years, and puts forward the future directions and key tasks of those diseases, in order to provide reference for accelerating the control and elimination programmes on key parasitic diseases in China.

Objective To investigate the effects of soil-transmitted nematode infection of mother and child on wheezing, asthma, and skin prick test (SPT) positive in rural preschool children. Methods Preschool children and their mohers in rural area were enrolled respectively, who underwent physical examination in the Yuncheng Central Hospital outpatient sector from April 2020 to March 2021. Basic information of mothers and children and children’s wheezing/asthma condition were collected through questionnaires. Fecal samples were collected from participants, and examined for soil-transmitted nematode eggs by the Kato-Katz thick smaear method, The SPT test was performed for children to collect allergic and atopic data. Multivariate unconditional logistic regression analysis was used to explore the relevant factors affecting asthma, wheezing, and SPT positivity in children. Results In this study, there were 2 014 children and their mothers each. The infection rate of soil-transmitted nematodes among the mothers was 14.40% (290/2 014), and among the preschool children was 7.99% (161/2 014). There were 117 children had wheezing symptoms, with an occurrence rate of 5.81% (117/2 014); 149 cases had asthma, with an occurrence rate of 7.40% (149/2 014) and 304 cases were positive for SPT, with a positive rate of 15.09% (304/2 014). Premature infants, mothers with a history of allergies, and family members smoking are risk factors for wheezing in children (P < 0.05, OR > 1); female gender and complete breastfeeding are protective factors for wheezing in children (P < 0.05, OR < 1). Mothers with a history of allergies, having someone smoking at home, and planting flowers and plants at home are risk factors for childhood asthma (P < 0.05, OR > 1); female and complete breastfeeding are protective factors for childhood asthma (P < 0.05, OR < 1). Mothers with a history of allergies is a risk factor for SPT positivity in children (P < 0.05, OR > 1), while the gender of the child, presence of soil-transmitted nematode infection in the child, and presence of soil-transmitted nematode infection in the mother are protective factors for SPT positivity in children (P < 0.05, OR < 1). Conclusion Mothers and children infected with soil-transmitted nematode may reduce the occurrence of allergic diseases in children.

Antimalarial drug resistance presents the biggest challenge for treatment against malaria. Plasmodium parasites have developed different degrees of resistance to common traditional antimalarial drugs including artemisinin. Therefore, the improvement of traditional drugs and the research and development of new drugs are urgently needed. This paper discusses the malaria control strategies based on a systematic review of the resistance mechanisms against traditional antimalarial drugs, the improvement strategies and optimisation achievements based on traditional drugs, and the research advances of new antimalarial drugs.

Objective To study the function of Anopheles stephensi peptidoglycan recognition protein S2 (PGRP-S2) and its role in regulating the homeostasis of symbiotic microbiota. Methods The female An. stephensi mosquitoes aged 2-4 days after eclosion were divided into control group (by biting the 10% sucrose solution), infected blood group (by biting the Plasmodium berghei ANKA-infected mice with 4%-8% parasitemia) and healthy blood group (by biting healthy mice), 60 mosquitoes in each group, the midgut and carcass were isolated after 24 h of full blood meal. The RNA was extracted by TRIzol, and relative transcription level of the pgrp-s2 gene was detected by real-time fluorescence quantitative PCR (RT-qPCR). Female An. stephensi mosquitoes 0-1 day after eclosion were divided into control group (fed with 10% sucrose solution) and antibiotic treatment group (fed with 10% sucrose solution containing penicillin, streptomycin and gentamicin), 30 mosquitoes in each group, midgut and carcass were isolated after fed for 5 days to detect the relative transcription level of pgrp-s2 gene by RT-qPCR. Female An. stephensi mosquitoes were divided into pgrp-s2 knockdown group and green fluorescent protein (gfp) control group, 60 mosquitoes in each group, each group were injected with pgrp-s2 double-stranded RNA (dsRNA) or green fluorescent protein gene dsRNA (69 nl/mosquito) respectively. After 2 days, mosquito RNA was extracted and RT-qPCR was used to detect the relative transcription level of pgrp-s2, mosquito DNA was extracted and PCR was used to detect the total amount of symbiotic bacteria in the mosquitoes using 16S rRNA specific primers. RNA was extracted from 30 An. stephensi mosquitoes of the knockdown group and control group respectively, which were fed with cotton balls soaked with 10⁷/ml Morganella morganii for 5 days, and subsequently, the relative total amount of 16S rRNA specific symbiotic bacteria and the relative number of M. morganii were detected by RT-qPCR; the RNA extracted from the midgut tissues of the knockdown group and gfp control group mosquitoes was use for transcriptome sequencing, cluster analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation and enrichment analysis. The PGRP-S2 amino acid sequence was predicted and analyzed using SMART website, and aligned using CLC Main Workbench software. The pgrp-s2 gene was cloned from An. stephensi mosquito cDNA, and the PGRP-S2 recombinant protein was expressed in SF9 cells using the insect baculovirus expression system (pFastbacI), and the protein expression was verified by Western blotting. The purified PGRP-S2 recombinant protein solution of 10, 20 and 40 μg/ml (protein buffer as control) was separately incubated with 40 μg of Lys-type peptidoglycan or DAP-type peptidoglycan, and the relative absorbance value (A₅₄₀) was detected every 12 h to check the degradation of peptidoglycan and verify the amidase activity of PGRP-S2. Results The RT-qPCR results showed that 24 h after the blood meal, the An. stephensi mosquitoes midgut pgrp-s2 relative transcription levels in the infected blood group, healthy blood group and control group were 1 590.0 ± 665.2, 126.8 ± 100.4 and 15.84 ± 6.92, respectively. The pgrp-s2 relative transcription levels of the infected blood group was higher than that of the control group (t = 2.38, P < 0.05); the pgrp-s2 of An. stephensi midgut relative transcription level of the control group was higher than that of the carcass (1.71 ± 0.51) (t = 2.04, P < 0.05). The pgrp-s2 of An. stephensi midgut relative transcription level of antibiotic treatment group was 0.33 ± 0.18, which was lower than that of the control group (117.9 ± 54.5) (t = 2.16, P < 0.05). The relative total amount of 16S rRNA of symbiotic bacteria in the pgrp-s2 knockdown group was 3 653 ± 2 023, which was lower than 14 982 ± 3 892 in the gfp control group (t = 2.58, P < 0.05); after feeding with M. morganii, the relative number of M. morganii in the An. stephensi of pgrp-s2 knockdown group was 571 517 ± 61 258, which was lower than 919 754 ± 123 397 of GFP control group (t = 2.53, P < 0.05). Transcriptome analysis results showed that pgrp-s2 knockdown could up-regulate An. stephensi Toll/Imd pathway, mTOR pathway, FoxO pathway and immune-related genes, while down-regulate metabolism-related genes such as fatty acid metabolism and tricarboxylic acid cycle. After incubation of PGRP-S2 recombinant protein at concentrations of 10, 20 and 40 μg/ml with DAP-type peptidoglycan for 48 h, the relative A₅₄₀ values were 0.49 ± 0.07, 0.40 ± 0.10 and 0.44 ± 0.07, respectively, which were lower than 0.90 ± 0.09 of control (t = 3.53, 3.65, 3.97; all P < 0.05); after incubation with Lys-type peptidoglycan for 48 h, the relative A₅₄₀ was 0.52 ± 0.03, 0.62 ± 0.03 and 0.65 ± 0.04, respectively, and there was no difference from 0.64 ± 0.05 of control (t = 1.95, 0.31, 0.11; all P > 0.05). Conclusion pgrp-s2 is mainly expressed in the midgut of An. stephensi, and the expression level is regulated by symbiotic bacteria. PGRP-S2 recombinant protein can degrade DAP-type peptidoglycan, having amidase activity, and may regulate the level of symbiotic bacteria in An. stephensi by negatively regulating immune responses and regulating metabolic responses.

The medical records of 4 cases of imported coronavirus disease 2019 (COVID-19) complicated with severe falciparum malaria diagnosed in Kunming Third People's Hospital in August 2022 were collected. The epidemiology, clinical manifestations, laboratory and imaging examinations, course of treatment and prognosis of 4 cases were analysed retrospectively. All the 4 cases were male, aged 40-54 years, and had a history of work and life in Africa. All cases had acute onset of symptoms, including fever (4 cases), chills (3 cases), shivering (3 cases), nausea and vomiting (3 cases), diarrhea (4 cases), and poor appetite (4 cases) as the main clinical manifestations. Headache, dizziness, unclear consciousness were reported in 2 cases, urinary incontinence in 1 case, muscle soreness in 2 cases, cough in 2 cases, phlegm in 1 case, and pharyngalgia in 1 case. All cases were positive for coronavirus. Nucleic acid test by throat swabs, and Plasmodium falciparum was found by microscopy examination of peripheral blood smear. All cases met the diagnostic criteria for severe malaria and were accompanied by abnormal liver function and severe hypoproteinemia, 2 cases of hyperbilirubinemia, 3 cases of dyslipidemia, 3 cases of abnormal blood picture involving the third system, increased calcitonin to varying degrees, 2 cases of lactic acid acidosis, and 1 case of hypoglycemia. Chest CT showed changes of viral pneumonia in 1 case. Antiviral drug and artesunate (intravenous injection 120 mg/time, once at 0, 12 and 24 hours, and once a day later for 7 consecutive days) were given, and different schemes of precise treatment were given based on the condition of the patient. All the patients were cured and discharged, and the follow-up was stable. It is suggested that the awareness of COVID-19 and malaria co-prevention should be established among the entry personnel in the high malaria epidemic areas abroad, to avoid serious cases and deaths.

Objective To understand the different ultrasonic manifestations of hepatic alveolar echinococcosis (HAE), to provide a reliable basis for improving the detection and diagnostic accuracy of HAE under conventional ultrasonography. Methods From January 2018 to April 2021, the patients who underwent routine ultrasound examination in the First Affiliated Hospital of Xinjiang Medical University and were diagnosed as HAE through surgery and pathology were recruited in the study. The relevant data, including gender, age, nationality, and ultrasonic image classification were collected. The clearest and complete images of HAE lesions in the original ultrasound images of the study subjects were selected for analysis, and the ultrasound manifestations of the lesions (location, size, boundary and shape, solid portion echo, calcification, liquefaction necrosis, blood flow signals, bile ducts and vascular invasion, etc.) were recorded, and values were assigned. Multiple lesions were described separately. The coincidence between post-operation pathological findings and the ultrasound diagnosis of the lesion was determined. Univariate analysis was conducted on the various ultrasound manifestations of HAE lesions, with statistically significant factors set as independent variables, while the coincidence with ultrasound diagnosis of lesions as dependent variables. A multivariate logistic regression analysis was conducted to establish a regression model, which was validated with the receiver operating characteristic (ROC) curve. The nomogram was constructed based on the results screened using multivariate logistic regression analysis, and its performance was evaluated using the ROC curve, calibration curve and decision analysis curve. Results The total number of HAE patients included in this study was 141. The age ranged from 9 to 65 years old, with an average age of (37.4 ± 13.6) years. There were 71 males and 70 females in this study, including 87 Tibetan patients accounting for 61.7%. Among the 141 HAE patients, 28 had single lesions, and 113 had multiple lesions. The final number of HAE lesions with inclusive criteria was 170. The results of univariate analysis showed that the difference in lesion location was not statistically significant (χ² = 1.952, P > 0.05). The differences in lesion size, boundary and shape, internal echo, calcification, liquefaction necrosis, blood flow signal, and bile ducts and vascular invasion were statistically significant (χ² = 39.026, 18.601, 15.743, 47.205, 34.151, 6.597, 21.766, all P < 0.05). The 4 factors including lesion size (OR = 0.180, 95% CI: 0.020-1.645), calcification (OR = 0.037, 95% CI: 0.002-0.590), liquefaction necrosis (OR = 0.282, 95% CI: 0.042-1.867), and blood flow signal (OR = 20.746, 95% CI: 3.720-115.686) were correlated with the ultrasound diagnosis compliance of HAE lesions. The accuracy analysis results of the logistic regression model predicted that the sensitivity of the regression model was 96.7% (118/122), the specificity was 83.3% (40/48), the positive predictive value was 93.7% (118/126) and the negative predictive value was 90.9% (40/44). The area under the ROC curve was 0.918 (95% CI: 0.859-0.977), the sensitivity was 95.1% and the specificity was 85.4%. The area under the ROC curve of the nomogram constructed based on the results of multivariate logistic regression analysis was 0.833. The sensitivity was 86.9% and the specificity was 70.8%. The calibration curve was close to the reference curve, and the decision analysis curve deviated significantly from both reference curves. Conclusion The size, calcification, liquefaction necrosis, and blood flow signal of the HAE lesions were important factors affecting ultrasound diagnosis.