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Objective To investigate the prevalence and molecular features of Cryptosporidium in captive-bred Mustela putorius furo in Jiangsu Province.. Methods A total of 290 fresh stool samples were collected from a ferret farm in Jiangsu Province on May 2017, and the small subunit rRNA (SSU rRNA) gene of Cryptosporidium was amplified in stool samples using nested PCR assay. The actin, cowp and gp60 genes were amplified in positive samples and sequenced to characterize Cryptosporidium species/genotypes. Results A total of 18 stool samples were tested positive for Cryptosporidium SSU rRNA gene, with a detection rate of 6.2%. Sequence and phylogenetic analyses of SSU rRNA, actin and cowp genes characterized Cryptosporidium isolated from captive-bred ferrets as Cryptosporidium sp. ferret genotype. In addition, gp60 gene was amplified in 10 out of 18 stool samples tested positive for Cryptosporidium. Conclusions Cryptosporidium is widely prevalent in captive-bred ferrets in Jiangsu Province, and Cryptosporidium sp. ferret genotype is the only Cryptosporidium genotype in ferrets.
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Purpose@#The purpose of the current study was to explore the functions and potential mechanism of miR-451a in breast cancer (BC). @*Methods@#Quantitative reverse transcription real-time polymerase chain reaction was used to analyze the expression of miR-451a in human normal mammary cells (MCF-10A) and BC cells. Colony formation assay, terminal-deoxynucleoitidyl transferase mediated nick end labeling assay and transwell assays were conducted to validate the effect of miR-451a on proliferation, apoptosis, migration and invasion of BC cells, respectively. RNA pull-down, RNA immunoprecipitation and luciferase reporter assays were applied to investigate the upstream and downstream mechanisms of miR-451a in BC cells. @*Results@#MiR-451a was expressed at a low level in BC cells. Overexpression of miR-451a repressed BC cells proliferation, migration and invasion. Moreover, long non-coding RNA AC092127.1 acted as a sponge of miR-451a to enhance the expression level of AE binding protein 2 (AEBP2) that was demonstrated to be the target gene of miR-451a in BC cells. Finally, rescue experiments validated that miR-451a and AEBP2 involved in AC092127.1-mediated BC cell growth, migration and invasion. @*Conclusion@#In a word, AC092127.1/miR-451a/AEBP2 axis contributes to BC cell growth, migration and invasion. Our results may help to find novel potential targets for BC treatment.
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Purpose@#The purpose of the current study was to explore the functions and potential mechanism of miR-451a in breast cancer (BC). @*Methods@#Quantitative reverse transcription real-time polymerase chain reaction was used to analyze the expression of miR-451a in human normal mammary cells (MCF-10A) and BC cells. Colony formation assay, terminal-deoxynucleoitidyl transferase mediated nick end labeling assay and transwell assays were conducted to validate the effect of miR-451a on proliferation, apoptosis, migration and invasion of BC cells, respectively. RNA pull-down, RNA immunoprecipitation and luciferase reporter assays were applied to investigate the upstream and downstream mechanisms of miR-451a in BC cells. @*Results@#MiR-451a was expressed at a low level in BC cells. Overexpression of miR-451a repressed BC cells proliferation, migration and invasion. Moreover, long non-coding RNA AC092127.1 acted as a sponge of miR-451a to enhance the expression level of AE binding protein 2 (AEBP2) that was demonstrated to be the target gene of miR-451a in BC cells. Finally, rescue experiments validated that miR-451a and AEBP2 involved in AC092127.1-mediated BC cell growth, migration and invasion. @*Conclusion@#In a word, AC092127.1/miR-451a/AEBP2 axis contributes to BC cell growth, migration and invasion. Our results may help to find novel potential targets for BC treatment.
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@#Objective To identify the genotype of Toxoplasma gondii isolated strains from a congenital teras（KS strain）and an HIV⁃Toxoplasma co⁃infected patient in Jiangsu Province. Methods T. gondii DNA of tachyzoites of a isolate from a congenital teras（KS strain）and blood DNA of an HIV⁃Toxoplasma co⁃infected patient in Jiangsu Province were extracted，and 11 loci were identified for the genotype by polymerase chain reaction restriction fragment length polymorphism（PCR⁃RFLP）. Results The complete bands were obtained from the congenital teras（KS strain）and HIV⁃Toxoplasma co⁃infected patient in Jiangsu Province，and identified as T. gondii gene type I. Conclusion T. gondii gene type I may be the dominant genotype strain of T. gondii among the women who have the abnormal pregnant outcomes and HIV⁃Toxoplasma co⁃infected patients in Jiangsu Province.
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Objective To investigate the expression and effect of high mobility group box 1(HMGB1) and its signaling pathway(HMGB1 RAGE/TLR4-NF-κB-cytokines)in rats with dilatd cardiomyopathy(DCM).Methods The rats were divided into two groups:normal control group (control,n=20) which treated with physiological saline,and DCM group(n=22) which treated with adriamycin(1 mg/kg twice a week)for 6 weeks,and then observed for 2 weeks.Echocardiography was performed at the end of the study.Plasma IL-1,IL-6,TNF-α level were measured by the flow cytometry.The CRP,BNP concentrations were measured by enzyme linked immunosorbent assay (ELISA).The expression of HMGB1 mRNA,TLR4 mRNA,RAGE mRNA,NF κB mRNA were measured by real time PCR.Results There were four rats dead in the DCM group;two rats were randomly selected from the DCM group to certified modeled successfully by echocardiography and pathological examination.Left ventricular end-diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) in DCM group were significantly higher than those in the normal control group(P＜0.05);left ventricular ejection fraction (LVEF),left ventricular short axis contractility(FS) in DCM group was significantly lower than that in normal control group(P＜0.05).The expression of H MGB1 mRNA,TLR4 mRNA,RAGE mRNA and NF-κB mRNA in myocardial tissue were significantly increased in DCM group than in the normal control saline group (P＜ 0.05),The expression of HMGB1 mRNA were positively correlated with TLR4 mRNA,RAGE mRNA and NF κB mRNA(r=0.873,P=0.005;r=0.949;P=0.000;r=0.898,P=0.002).The serum levels of IL-1,IL 6,TNF α and CRP were significantly higher in DCM group.The expression of HMGB1 mRNA in myocardial tissue was positively correlated with IL 1,IL-6,TNF-α and CRP(r=0.944,P=0.002;r=0.988,P=0.000;r=0.968,P=0.000;r=0.961,P=0.000).Conclusion HMGB1 and it's inflammation signaling pathway (HMGB1-TLR4/RAGE-NF-κB-cytokines) were highly expressed in dilated cardiomyopathy,and have relationship with left ventricular diameter and cardiac function,they may be involved in the development of DCM.
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Objective: To explore the application of18F-lfuorodeoxyglucose (FDG) micro- positron emission tomography (PET) myocardial metabolism imaging for evaluating dilated cardiomyopathy model (DCM) in experimental rats. Methods: A total of 12 male SD rats were randomly divided into 2 groups: DCM group, the rats received intraperitoneal injection of adriamycin at 1.0 mg/kg twice per week and Control group, the rats received intraperitoneal injection of normal saline, all animals were treated for 6 weeks followed by 2 weeks observation.n=6 in each group. Echocardiography was performed at pre- and post-modeling,18F-FDG micro-PET myocardial metabolism imaging was conducted after modeling and plasma level of BNP was examined as well. Finally, the rats were scariifed to observe the pathological changes of myocardial tissue. Results: 1 rat died in DCM group and the rest were with successful modeling conifrmed by echocardiography and pathology. Compared with Control group, DCM group showed decreased standard uptake value of18F-FDG (1.23 ± 0.55) vs (6.65 ± 0.41),P<0.01; the standard uptake value of18F-FDG was negatively related to left ventricular end diastolic diameter (LVEDD) (R=-0.709,P=0.015), LVESD (R=-0.924, P=0.000) and plasma level of BNP (R=-0.948,P=0.000), while positively related to LVEF (R=0.968,P=0.000) and fractional shortening (R=0.863,P=0.001). Conclusion:18F-FDG micro-PET myocardial metabolism imaging combining echocardiography, biochemical and pathological examinations may evaluate DCM modeling in rats, which provide a non-invasive and intravital tool for small animal experiment.
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Objective To observe the effect of artificially construction of the life cycle of Angiostrongylus cantonensis in the laboratory condition so as to provide the basis for the research of angiostrongyliasis. Methods SD rats were infected orally with the third?stage larvae of A. cantonensis collected from infected Pomacea canaliculata. Six weeks after the infection the first?stage larvae were isolated and counted from fresh feces of the rats and then were used to infect P. canaliculata. Three weeks lat?er the snails were dissected for counting the third?staged larvae of A. cantonensis. Results The first?stage larvae were detect?ed in the feces of the rats 6 weeks after the infection and the third?staged larvae were successfully isolated after the infection of P. canaliculata. Conclusion The animal model of the entire life cycle of A. cantonensis is successfully established in the labo?ratory with the infection of 50 larvae per rat.
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Objective To discuss the significance of region Ⅵ lymph nodes dissection in treatment of patients with clinical cervical nodes negative(cN0)papillary thyroid carcinoma(PTC).Methods Clinical data of 38 cases of cN0 PTC treated with region Ⅵ lymph nodes dissection were retrospectively analyzed.The related literature was reviewed.The cervical nodes dissection scope and the key operation points in treatment of cN0 PTC were discussed.Results No permanent recurrent laryngeal nerve injury or parathyroid damage happened.14 cases(36.84％)had occult lymph node metastasis to region Ⅵ lymph nodes.After more than 5 years of follow-up,all the cases had excellent life quality.3 cases(7.89％)were found lymph node metastasis to lateral cervical region and they were given functional neck dissection.No recurrence or metastasis occurred to the 3 cases during more than 2 years of follow-up.Conclusion Region Ⅵ lymph node dissection in treatment of cN0 PTC is safe,reliable,and with less complications.
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Objective To study the effects of lipopolysaccharide(LPS) on the expression of toll like receptor 4 (TLR4), reactive oxygen species(ROS) and on the proliferation of cells as well as secretion of six proinflammmatory cytokines including TNF-α, IL-1, IL-6, IL-8, IL-10, IL-12 levels in SKOV3 cells. And to explore the mechanism of SKOV3 cells in regulation. Methods Cultured primary SKOV3 cells were stimulated with different concentrations of LPS (0.01 μg/ml, 0.1 μg/ml, 1 μg/ml, 10 μg/ml and 20 μg/ml) for 4 h, the TLR4 expression in SKOV3 cells were examined by flow cytometry;1 μg/ml LPS stimulated SKOV3 for 4 h, 8 h, 12 h, 24 h respectively, the TLR4 expression and cell cycle in SKOV3, cell proliferation, ROS level as well as cells and TNF-α and IL-1, IL-6, IL-8, IL-10, IL-12 levels in the culture medium were assayed by flow cytometry, MTT, CBA assay respectively. Results LPS with different concentrations of LPS stimulation in-duced an increased TLR4 expression, however, the expression was reduced when LPS concentration up to 10 μg/ml. LPS stimulation for 4 h, 8 h induced an increased TLR4 expression and cell proliferation. Stimulated for 24 h, however, the TLR4 expression and cell growth were inhibited in S period. Meanwhile, LPS stimulation for 4 h, 8 h, 12 h, 24 h induced a higher ROS secretion in comparison with control group. LPS stimulation induced a stronger cytokine response in comparison with control group, as demonstrated by the production of TNF-α, IL-1, IL-6, IL-8 secretion in cultured SKOV3 cells, while IL-10 and IL-12 with low expression have no obvious difference in the all medium samples. Conclusion TLR4 expression, cell proliferation, ROS and proin-flammmatory cytokine secretion could be induced in SKOV3 through LPS stimulation. The study provide new ex-periment evidences for human ovarian cells SKOV3 immunity regulation and inflammation reaction to promote cells inhibition after LPS stimulation.
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Objective To explore the impact of acute Toxoplasma gondii infection on cerebral proteins and nerve growth in mice by 2D electrophoresis. Methods The cerebral proteins from C57BL/6J mice infected with Toxoplasma gondii and normal paired mice were extracted. The discrepant proteins were checked by 2D electrophoresis. Isoelectric focusing was determined as the first direction (immobilized pH gradient gel 3-10) ,SDS-PAGE as the second direction to execute 2D electrophoresis, and PDQuest 1.0 software was used to analyze 2D electrophoretogram. Results The protein spots in Toxoplasma gondii infected mice and normal paired mice were (132 ±10) and (170 ± 13) , respectively. After the analysis by PDQuest 1. 0 software, only 19 protein spots were found to express in infected mice and only 37 protein spots were found to express in normal paired mice. Additionally, the obvious quantitative changes in a part of proteins of the cerebrum in the both group occurred. Conclusion There are obvious changes in cerebral proteins from mice with acute Toxoplasma gondii infection, which provids useful clues for studying the cerebral proteins injury in acute Toxoplasma gondii infected mice and the new cure drug.
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Objective To study the male reproductive ability of male rats with Toxoplasma gondii ( Tg) infection and investigate the variation of Toxoplasma infection in seminal plasma of infertile patients and explore its mechanism. Methods Thirty SD rats were randomly divided into 3 groups. The rats in the Toxoplasma infection group were administrated intraperitoneally with tachyzoites of Tg. in a dosage of 2 × 10~ 5/ml(2ml) , the rats in the treated group were administered with the same dosage of the tachyzoites and from the second day after the infection they were treated with 200 mg/kg azithromycin for 7 days, and the normal group was given physiological saline. Nine weeks after the infection, the serum sex hormone level, number,vitality, activity and quantity of spermatozoa and activities of enzymes in testa's of the testicular tissues were determined in the male rats. The female rats infected with Tg were matched with normal female rats at a ratio of 1: 2 for one week, and on the 21st day of pregnancy, the number of corpora luteum, sex ratio and the weight, body length and tail length of fetus were measured. The ELISA method was used todetermine the seminal plasma's anti-Tg IgG antibody of the 169 patients with infertility and 35 males with normal fertility. Meanwhile the NO levels in their semina were determined by means of nitric acid reducase. Results The number, activity .vitality, serum level of sex hormones were all lower in the infected rats than those in the normal and treated groups. The number of fetus in the pregnant rats matched with the infected male rats was significantly fewer, but the average body weight, body length, tail length of the fetuses and sex proportion showed no significant difference in comparison with those of the control group. The anti-Toxoplasma gondii antibody positive rate in the masculine infertility patients was 18.35% , being significantly higher than 2. 86% in the normal fertility group(P < 0.05 ). The mean NO level in the semina from the infertility group was (146.68 ± 38. 87) μnol/L , which was significantly higher than (84.92 ± 26.72) μnol/L( P < 0.01) in the fertility group. Conclusion Toxoplasma gondii infection can cause certain influences on the male reproductive ability.
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To investigate the effect of Toxoplasma gondii infection upon the expression of brain-derived neurotrophic factor (BDNF) mRNA and N-methyl-D-aspirate receptor (NMDA) subunits NR2A and NR2B,Wistar rats of 4 weeks old were randomly divided into 3 groups with 10 rats in each group in which 2 mL suspensions of T.gondii tachyzoits in the concentrations of 2×10~7/mL and 2×10~5/mL were injected intra-peritoneally to rats in group A and group B respectively, serving as the experimental groups, while 2 mL of sterile physiologic saline was injected intra-peritoneally in group C serving as the control group. Four weeks after injection, the expressions of BDNF mRNA and BDNF protein in the brain tissues were detected by in situ hybridization and immunohistochemical assay and the expressions of NR2A and NR2B immune activity in the hippocampal CA1,CA3 and DG were investigated by using computer-assisted image analysis system. Compared with the control group, the expression of BDNF protein in the hippocampus of the experimental groups was significantly enhanced [(64.27±23.18), (50.39±19.34) vs (44.68±22.74)/mm~2,P＜0.05]. In addition, the increased expressions of BDNF mRNA in the hippocampus of the experimental groups were also demonstrated [(0.13±0.02), (0.12±0.02) vs (0.09±0.01); P＜0.05]. In the expression of the NR2A protein, their expressions in group A and B of rats were significantly lower than that of group C in CA3 (P＜0.05),but there was no significant change in CA1 and DG. In the expression of NR2B protein, the expressions in group A and B were also lower than that of group C in CA1 and CA3, and had no significant change in DG. It is evident that the expressions of BDNF mRNA and BDNF protein in hippocampal tissues were significantly increased following chronic infection with T.gondii, supporting the hypothesis that BDNF may be involved in the intrinsic neuro-protective mechanism.
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Objective To search for a biomarker from the serum of Schistosoma japonicum infected rabbits for early diagnosis of schistosomiasis. Methods The sera were obtained from different periods of the infected rabbits. The serum proteins were generated by WCX kit (Bruker Daltonics GmBH) and analyzed by the technique of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Results In the mass range from 1 000 to 12 000 Da, sixty-three proteins were captured by WCX kit. ClinProTools software was used to find the differential expressed proteins. The result revealed 7 distinct proteins compared with normal serum. Among them,1 787 Da protein expression was increased (P
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Objective To explore the relationship between tumor necrosis factor-?(TNF-?)level in the serum of maternal women with intrauterine infection of Toxoplasma gondii and absortion.Methods An examination was carried out on TNF-? in maternal sera and DNA of Toxoplasma gondii in the cervical secretions,and the aborted tissues of the abortion group,and the cervical secretions of the control group by ELISA and PCR,respectively.Results The TNF-? levels increased in the sera of women with intrauterine infection of Toxoplasma before abortion as compared with those of the control group and those of themselves after abortions.There were high TNF-? levels in the sera of prognostication abortion women with Toxoplasma infection before the anti-Toxoplasma treatment as compared with those of the control group and those of themselves after the treatment with azithromycin.Conclusion The results suggest that Toxoplasma gondii infection is an important factor to increase the TNF-? level in maternal serum.
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Objective To research the therapeutic effect of dihydroarteminsinin/piperaquine phosphate(duo-cotecxin)on rats with Pneumocystis pneumonia(PCP),and explain its potential mechanism.Methods The PCP rat model was established through the subcutaneous injection with cortisone acetate twice a week for 6 weeks.The model rats were orally treated with duo-cotecxin at a dose of 40 mg/kg once a day for 3 days,and with cotecxin at a dose of 60 mg/kg once a day for 6 days,respectively.The therapy control group was treated orally with SMZco(sulfamethoxazol 250 mg/kg +Trimethoprim 50 mg/kg),positive control group and normal group were established,respectively.The drug efficacy was based on rat survival rate,increase rate of body weight,lung weight/body weight ratio,mean numbers of the cysts per field in the lung print smear,and the changes of CD4+ T cells,CD8+ T cells,NO,IFN-? and TNF-? in the blood.Results The survival rates and the body weight of rats treated with duo-cotecxin and cotecxin were higher and heavier than those of the positive control group respectively,which approached to those of the group treated with SMZco.The lung weight/body weight ratio,and mean cyst number per field of lung print smear of the rats treated with duo-cotecxin and cotecxin were lower than those of the positive control group,respectively.NO and TNF-? in the blood of the rats treated with duo-cotecxin and cotecxin were lower than those of the positive control group,respectively(P
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Objective To explore the influence of Toxoplasma tachyzoites infection on motility of human spermatozoa in vitro, and explain its possible mechanism. Methods Semen samples obtained from 10 healthy volunteers by masturbation were prepared by the swim-up technique. The samples were then inoculated at 37 ℃ with different concentrations of live Toxoplasma tachyzoites varying among 1?103/ml (Group A), 1?104/ml (Group B), 1?105/ml (Group C), 1?106/ml (Group D), 1?107/ml (Group E) and Group F containing Ham’s F-10 only as the negative control. Motion parameters were analysed by Computer-aided sperm analyzer(CASA) in 0 hour, 1 hour, 2 hours, 4 hours respectively. Modalities of spermatozoa and possible adherence and/or agglutination were observed under the light microscope. Results Distinct adhesion of spermatozoa to Toxoplasma tachyzoites and agglutination were noticed. In all the motion parameters, progressive motility was affected most and dependent upon the incubation time and tachyzoites concentration. Progressive motility showed a significant difference between Group E and the control (P
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Objective To determine the levels of anti-Toxoplasma gondii antibody (ATAb) and antisperm antibody (AsAb) in serum samples and interferon-?(IFN-?) in supernatants of peripheral blood cell culture in females with infertility, and to analyze their correlation. Methods ELISA was applied to determine and compare the ATAb and AsAb in serum samples of 182 women with infertility and those of 94 women with normal pregnant history. The level of IFN-? in supernatants of peripheral blood cell culture was also measured and compared by ELISA. Results The positive rates of serum ATAb and AsAb in the cases of infertility were 15.38% and 18.13% respectively, which were significantly higher than those of women with normal pregnant history( 6.38% and 5.32%). The supernatant IFN-? level in infertile women was significantly higher than that of women with normal pregnant history (0.970?0.493) ?g/L vs (0.531?0.274) ?g/L(P
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Objective To explore the effect of reducing sub-clinical infections of Toxoplasma gondii in newborn babies with preventive treatment. Methods Forty-four infected pregnant women were treated in different period and their umbilical blood and/or placenta of their newborn babies were tested for IgG, IgM, cAg and DNA of Toxoplasma gondii. Results The positive rate of Toxoplasma DNA in umbilical blood was 37.8%(14/37). The lowest rate was found in the preventive treatment group with a significant difference compared with the group of general treatment. Conclusion Preventive treatment can reduce sub-clinical infection rate of newborn babies effectively.
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Objective To inquire into the relationship between infections of Toxoplasma gondii and apoptosis of spermatogenic cells. Methods Apoptotic spermatogenic cells of mice infected with Toxoplasma gondii were examined by Wright-Giemsa staining and terminal deoxynucleotidyl trans-Icrase (TdT) mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labeling (TUNED techniques. Results Apoptosis rate of infective group of Toxoplasma was significantly higher than thai oi control group (F
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Objective To observe the changes of peripheral blood T lymphocytes,IFN-?,TNF-? and IL-4 in rats infected by T.gondii.Methods 48 Sprague-Dawley(SD)rats were intra-abdominally injected with 2?105/L of cellulose purified living tachyzoites in 2 ml and randomly divided into 8 groups.Six rat was intra-abdominally injected 2 ml of saline as control and 4 rats were remained as normal control.Peripheral blood was collected and the level of IFN-?,TNF-?,IL-4 was analyzed by ELISA on day 1,3,7,14,28,35,42,60.Results Level of IFN-?(6.73 pg/ml)and IL-4(6.91 pg/ml)increased in experimental rats on day 7(P