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Objective To investigate the prevalence and influencing factors of Blastocystis hominis infection among children with diarrhea under five years of age in Guangzhou City. Methods Children with diarrhea under 5 years of age admitted to Guangzhou Children’s hospital, Guangzhou Maternity and Child Healthcare Hospital and Guangzhou Women and Children’s Medical Center during the period between January 1 and December 31, 2020, were enrolled. Participants’ demographics, living environments and health status were collected using questionnaire surveys. Stool samples were collected from participants and nucleic acid was extracted. B. hominis infection was identified using PCR assay and sequence alignment, and the factors affecting B. hominis infection among children with diarrhea under 5 years of age were identified using univariate analysis and multivariate logistic regression analysis. Results A total of 684 children with diarrhea under 5 years of age were enrolled, including 468 male children and 216 female children, with a mean age of (1.79 ± 1.12) years. The overall prevalence of B. hominis infection was 4.97% [34/684, 95% confidential interval (CI): (3.59%, 6.86%)] among participants, and there was no significant difference in the prevalence of B. hominis infection between children with chronic [7.52% (20/266), 95% CI: (4.92%, 11.33%)] and acute diarrhea [3.35% (14/418), 95% CI: (2.01%, 5.54%)] (χ2 = 5.983, P = 0.014). Multivariate logistic regression analysis identified keeping pet [odds ratio (OR) = 6.298, 95% CI: (2.711, 14.633)], drinking non-tap water [OR = 4.522, 95% CI: (1.769, 11.561)], lactose intolerance [OR = 4.221, 95% CI: (1.043, 17.087)], antibiotic use [OR = 0.125, 95% CI: (0.017, 0.944)] and chronic diarrhea [OR = 2.172, 95% CI: (1.018, 4.637)] as factors affecting B. hominis infection among children with diarrhea under 5 years of age in Guangzhou City. Conclusions B. hominis infections is detected in children with diarrhea under five years of age in Guangzhou City. Improving home environments and pet-keeping hygiene is recommended to reduce the likelihood of B. hominis infection among children.
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Objective To evaluate the immunoprotective effect of active immunization with recombinant peptidyl-prolyl cis-trans isomerase from Babesia microti against B. microti infection in mice. Methods Female BALB/c mice at 6 weeks of age, each weighing approximately 20 g, were divided into the recombinant protein immunization group, the infection control group and the normal control group, of 25, 18, 15 mice in each group, respectively. Mice in the recombinant protein immunization group were given active immunization with recombinant BmPPIase protein, and 18 mice with the highest antibody titers were intraperitoneally injected with 100 μL of B. microti-infected whole blood 2 weeks after the last immunization. Mice in the infection control group were intraperitoneally injected with 100 μL of B. microti-infected whole blood, while 15 mice in the normal control group received no treatment. Blood samples were collected from mice in the recombinant protein immunization group and the infection control group on days 0 to 30 post-immunization for detection of B. microti infection, and blood samples were collected on days 0, 7, 14, 21, and 28 post-immunization for routine blood tests with a blood cell analyzer and for detection of serum cytokines using cytometric bead array. Results Anti-BmPPIase antibodies were detected in 25 mice in the recombinant protein immunization group 2 weeks after the last immunization, with titers of 5 × 103 to 8 × 104. B. microti infection rate peaked in mice in both the recombinant protein immunization and the infection control group on day 7 post-immunization, with positive infection rates of 13.3% and 50.0%, and there were significant differences between the two groups in terms of B. microti infection rate on days 3 (χ2= 113.18, P < 0.01), 5 (χ2 = 475.22, P < 0.01), 7 (χ2 = 465.98, P < 0.01) and 9 post-infection (χ2= 18.71, P < 0.01), while the B. microti infection rate tended to be 0 in both groups on day 11 post-immunization. Routine blood tests showed higher red blood cell counts [(5.30 ± 0.50) × 1012 to (9.87 ± 0.24) × 1012 counts/L)] and hemoglobin levels [(89.67 ± 22.80) to (148.60 ± 3.05) g/L)] in the recombinant protein immunization group than in the infection control group on days 0 to 28 post-immunization. Cytometric bead array detected higher serum interferon-γ [(748.59 ± 17.56) to (3 858.28 ± 1 049.10) fg/mL], tumor necrosis factor-α [(6 687.34 ± 1 016.64) to (12 708.13 ± 1 629.79) fg/mL], interleukin (IL)-6 [(611.05 ± 75.60) to (6 852.68 ± 1 554.00) fg/mL] and IL-17a [(167.68 ± 185.00) to (10 849.27 ± 355.40) fg/mL] and lower IL-10 levels [(247.65 ± 138.00) to (18 787.20 ± 2 830.22) fg/mL] in the recombinant protein immunization group than in the infection control group during the study period. Conclusions Recombinant BmPPIase protein induces up-regulation of interferon-γ, tumor necrosis factor-α and presents a high immunoprotective activity against B. microti infection in mice, which is a potential vaccine candidate protein.
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Echinococcosis is a zoonotic parasitic disease caused by Echinococcus infections, and this disorder may cause fibrosis of multiple vital organs, which may further progress into cirrhosis. Early-stage hepatic fibrosis is reversible, and unraveling the mechanisms underlying hepatic fibrosis induced by Echinococcus infections is of great significance for the prevention and treatment of early-stage hepatic fibrosis. Recently, the studies pertaining to hepatic fibrosis associated with Echinococcus infections focus on cytokines and immune cells. This review summarizes the advances in the mechanisms underlying host immune cells- and cytokines-mediated hepatic fibrosis in humans or mice following Echinococcus infections.
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OBJECTIVE@#Depression and metabolic disorders have overlapping psychosocial and pathophysiological causes. Current research is focused on the possible role of adiponectin in regulating common biological mechanisms. Xiaoyao San (XYS), a classic Chinese medicine compound, has been widely used in the treatment of depression and can alleviate metabolic disorders such as lipid or glucose metabolism disorders. However, the ability of XYS to ameliorate depression-like behavior as well as metabolic dysfunction in mice and the underlying mechanisms are unclear.@*METHODS@#An in vivo animal model of depression was established by chronic social defeat stress (CSDS). XYS and fluoxetine were administered by gavage to the drug intervention group. Depression-like behaviors were analyzed by the social interaction test, open field test, forced swim test, and elevated plus maze test. Glucose levels were measured using the oral glucose tolerance test. The involvement of certain molecules was validated by immunofluorescence, histopathology, and Western blotting. In vitro, hypothalamic primary neurons were exposed to high glucose to induce neuronal damage, and the neuroprotective effect of XYS was evaluated by cell counting kit-8 assay. Immunofluorescence and Western blotting were used to evaluate the influences of XYS on adiponectin receptor 1 (AdipoR1), adenosine 5'-monophosphate-activated protein kinase (AMPK), acetyl-coenzyme A carboxylase (ACC) and other related proteins.@*RESULTS@#XYS ameliorated CSDS-induced depression-like behaviors and glucose tolerance impairment in mice and increased the level of serum adiponectin. XYS also restored Nissl bodies in hypothalamic neurons in mice that exhibited depression-like behaviors and decreased the degree of neuronal morphological damage. In vivo and in vitro studies indicated that XYS increased the expression of AdipoR1 in hypothalamic neurons.@*CONCLUSION@#Adiponectin may be a key regulator linking depression and metabolic disorders; regulation of the hypothalamic AdipoR1/AMPK/ACC pathway plays an important role in treatment of depression by XYS.
Subject(s)
Animals , Mice , AMP-Activated Protein Kinases/metabolism , Acetyl-CoA Carboxylase/metabolism , Adiponectin/metabolism , Antidepressive Agents/pharmacology , China , Depression/drug therapy , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Glucose , Hypothalamus/metabolism , Receptors, Adiponectin/metabolismABSTRACT
Human sparganosis is a food-borne parasitic disease caused by the plerocercoids of Spirometra species. Clinical diagnosis of sparganosis is crucial for effective treatment, thus it is important to identify sensitive and specific antigens of plerocercoids. The aim of the current study was to identify and characterize the immunogenic proteins of Spirometra erinaceieuropaei plerocercoids that were recognized by patient sera. Crude soluble extract of the plerocercoids were separated using 2-dimensional gel electrophoresis coupled with immunoblot and mass spectrometry analysis. Based on immunoblotting patterns and mass spectrometry results, 8 antigenic proteins were identified from the plerocercoid. Among the proteins, cysteine protease protein might be developed as an antigen for diagnosis of sparganosis.
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Objective To construct a cDNA library of Sparganum mansoni and immunoscreen antigen candidates for immunodiagnosis of sparganosis mansoni. Methods Total RNA was extracted from S. mansoni, and reversely transcribed into cDNA, which was ligated into the phage vector. These recombinant vectors were packaged in vitro to construct the SMART cDNA library of S. mansoni. Then, the cDNA library was immunoscreened with sera from patients with sparganosis mansoni to yield positive clones. The inserted fragments of positive clones were sequenced and subjected to homology analyses, and the structure and functions of the coding proteins were predicted. Results The SMATR cDNA library of S. mansoni was successfully constructed. The titer of the cDNA library was 6.25 × 106 pfu/mL, with a recombinant efficiency of 100%, and the mean length of the inserted fragments in the library was larger than 1 100 bp. A total of 12 positive clones were obtained by immunoscreening, and were categorized into Sm-I (Sm60-1), Sm-II (Sm58-1), Sm-III (Sm20-1) and Sm-IV (Sm22-3), with 1 134, 1 063, 883 bp and 969 bp long inserted fragments. Their coding proteins were highly homologous with the Spirometra erinaceieuropaei antigenic polypeptide, cytoplasmic antigen, ribosomal protein S4-like protein and unnamed protein product, respectively. Conclusions A SMART cDNA library of S. mansoni has been successfully constructed and 4 categories of positive clones have been identified, which provides a basis for further studies on diagnostic antigens for sparganosis mansoni.
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OBJECTIVE: To explore the value of nerve trunk stimulation in the rehabilitation of lower limb function in the patients with cerebral apoplexy at convalescence stage. METHODS: According the random number table, the patients with the lower limb dysfunction of cerebral apoplexy at convalescence stage were divided into a control group and a treatment group, 42 cases in each group. The drug therapy and the routine rehabilitation training were provided in the two groups. Additionally, in the treatment group, the nerve trunk stimulation therapy was adopted， in which, Chize (LU5，stimulating point of radial nerve), Neiguan (PC6, stimulating point of median nerve), Xiaohai (SI8, stimulating point of ulnar nerve) were selected. In the control group, acupuncture intervention was supplemented. Before and after treatment, the peak torque (PT) of the lower flexor-extensor muscle of the knee joint, gait parameters，the score of the modified Ashworth spasm scale (MAS), the score of Fugl-Meyer motor assessment (FMA) and the score of Fugl-Meyer balance scale (FBS) were recorded. RESULTS: After the treatment, the PT of the lower flexor-extensor muscle of the knee joint，the scores of FMA and FBS，the step speed and frequency were all increased, the score of MAS and the difference in the stride between the left and the right were decreased as compared with those before treatment (P<0.01). After the treatment, The PT of the lower flexor-extensor muscle of the knee joint，the scores of FMA and FBS，the step speed and frequency in the treatment group were higher than those in the control group (P<0.01). The score of MAS and the difference in the stride between the left and the right in the treatment group were lower than those in the control group (P<0.01). CONCLUSION: Nerve trunk stimulation therapy quite effectively increases the muscle strength and relieves the muscle tension as well as improves the motor function, the balance and the walking pattern of the lower limbs. This therapy is significantly valuable in the rehabilitation of the lower limbs in the patients with cerebral apoplexy at convalescence stage.
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Objective To investigate the prevalence and risk factors of Blastocystis infections among primary school students in Jiangjin District, Chongqing City. Methods A cross-sectional questionnaire survey was conducted among students sampled from a primary school in Jiangjin District, Chongqing City on April, 2018, and their stool samples were collected for microscopic examinations, in vitro culture and PCR assays to analyze the prevalence of Blastocystis infections and subtype of the parasite. In addition, the risk factors of Blastocystis infections among primary school students were identified using univariate analysis and multivariate logistic regression analysis. Results A total of 466 primary students were surveyed, and the subjects had a mean age of (9.81±1.66) years and included 236 males (50.64%) and 230 females (49.36%). The prevalence of Blastocystis infections was 15.24% (71/466) among the study students, and there was no significance difference in the prevalence between male and fe- male students (16.52% vs. 13.91%; χ2 = 0.616, P = 0.433). In addition, there was a significant difference in the prevalence of Blastocystis infections among grade 1 (6.35%, 4/63), grade 2 (5.17%, 3/58), grade 3 (21.74%, 15/69), grade 4 (25.30%, 21/83), grade 5 (10.19%, 11/108) and grade 6 students (20.00%, 17/85) (χ2 = 15.410, P = 0.009). There were four Blastocystis subtypes characterized (ST1, ST3, ST6 and ST7), in which ST6 was the most common subtype (45.07%, 32/71), followed by ST3 (25.35%, 18/71). Multivariate logistic regression analysis revealed that minority ethnicity [odds ratio (OR) = 4.259, 95% confidential inter- val (CI) : (1.161, 15.621)] and low maternal education level (primary school and below) [OR = 9.038, 95% CI: (1.125, 72.642)] were identified as risk factors of Blastocystis infection among primary school students in Jiangjin District, Chongqing City. Conclusions There is a high prevalence of Blastocystis infections detected among primary school students in Jiangjin District, Chongqing City, and ST6 and ST3 are predominant subtypes. Minority ethnicity and low maternal education level (primary school and below) are risk factors for Blastocystis infections in primary school students.
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ObjectiveTo investigate the prevalence and risk factors of Blastocystis hominis infections among AIDS patients in Nanchang City. MethodsA cross-sectional questionnaire survey was conducted among AIDS patients in Nanchang City during the period between May and September, 2016. B. hominis infection was detected in patients’stool samples using a PCR assay, and the CD4+ T cell count was measured in subjects’blood samples. In addition, the risk factors of B. hominis infection in AIDS patients were identified using univariate and multivariate logistic regression analyses. Results A survey was conducted in Nanchang City from May to September 2016. A total of 505 AIDS patients were investigated, and the prevalence of B. hominis infection was 4.16%. Univariate analysis revealed that B. hominis infection correlated with the occupation (χ2 = 8.595, P = 0.049), education level (χ2 = 14.494, P = 0.001), type of daily drinking water (χ2 = 10.750, P = 0.020), root of HIV infections (χ2 = 8.755, P = 0.026) and receiving anti-HIV therapy (χ2 = 23.083, P = 0.001) among AIDS patients, and multivariate logistic regression analysis identified daily direct drinking of tap water as a risk factor of B. hominis infections [odds ratio (OR) = 7.988, 95% confidential interval (CI): (1.160, 55.004)] and anti-HIV therapy as a protective factor of B. hominis infection [OR = 0.183, 95% CI: (0.049, 0.685)]. Conclusions The prevalence of B. hominis is 4.16% among AIDS patients in Nanchang City. Daily direct drinking of tap water is a risk factor, and anti-HIV therapy is a protective factor of B. hominis infection among AIDS patients living in Nanchang City.
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Objective To evaluate the effects of intravenous injection of different blood components containing Babesia microti on B. microti infection in mice. Methods Healthy mice were infected with B. microti, and then blood samples were collected from the mouse orbit to prepare whole blood, serum-free blood components and pure red blood cells containing B. microti. Twenty seven BALB/c mice were divided into three groups, including the whole blood group, the serum-free blood component group and the pure red blood cell group, of 9 mice in each group, and then, each group was divided into three subgroups, of 3 mice in each subgroup, which were injected with 100 μL of blood components containing B. microti at concentrations of 9.00, 0.90, 0.09 B. microti parasites/μL (900, 90, 9 B. microti parasites) via the tail vein, respectively. Blood samples were collected from the mouse tail tip every other day since one day post-injection to prepare thin blood smears. Following Giemsa staining of blood smears, B. microti infection was identified in red blood cells using microscopy. Results Following injection of 900 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group and the serum-free blood component group 3 days post-injection, and the density of B. microti parasites started to increase 15 days post-injection and peaked 21 days post-injection, with 2.21% and 1.76% rates of B. microti infection in red blood cells, respectively. Subsequently, the density of B. microti parasites declined, and the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was found in the peripheral blood in the pure red blood cell group. Following injection of 90 B. microti parasites, B. microti was identified in the peripheral blood in the whole blood group 3 days post-injection, and the density of B. microti parasites increased 15 days post-injection and peaked 21 days post-injection, with a 1.35% rate of B. microti infection in red blood cells, while the percentage of B. microti infection in red blood cells tended to be 0 31 days post-injection. During the study period, no B. microti was detected in the peripheral blood in the serum-free blood component group or the pure red blood cell group. Following injection of 9 B. microti parasites, no B. microti was detected in the peripheral blood in the whole blood group, the serum-free blood component group or the pure red blood cell group. Conclusion Blood components and dose of B. microti parasites may affect intravenous injection of B. microti injection in mice, and transfusion of blood components may case a risk of Babesia infection.