Objective In order to investigate the influence of reelin deficiency on the hippocampal development, the histochemical characteristics of hippocampal pyramidal cells and granule cells of reeler mice were analyzed, therefore, reelin's function would be better understood with more morphological evidences. Methods With immunofluorescent double labeling, the pyramidal cells, granule cells and mossy cells in hippocampi between wild type and reeler mice were labeled. Results The development and lamination of hippocampal cortical plate were in disorder. Pyramidal cells and granule cells dispersed, and moreover, granule cells proliferated rapidly and migrated into hilus, so that the bound between granule layer and hilus disappeared. Conclusion As a stop signal and regulatory signal, reelin plays important roles in neuronal migration of developing cortical plate, especially in the regulation of granule cell proliferation.

Objective In order to make a epileptic model in organotypic slice culture and to further investigate the epileptic pathology. Methods Immunohistochemistry, Fluoro-Jade B staining and BrdU labeling were carried out in the hippocampal slices with domoic acid treatment after 7 days of culture.Results Domoic acid could induce a series of pathological changes, such as, dispersion of granule cells(0.105±0.063) mm vs (0.057±0.012) mm, t = 4.8, P<0.01), neurogenesis in granular layer and cell loss of pyramidal cell and Mossy cell as well. Conclusions In hippocampal organotypic slices, domoic acid induced such pathological changes as human temporal lobe epilepsy and epileptic animal model. It is an ideal epileptic model for pathological, pharmacological and electronic physiological studies with great applicable prospect.

Objective To evaluate the effect of single-nucleotide polymorphisms at the miRNA binding site rs3660 in the 3'-untranslated region of the KRT81 gene (miR-SNPs) on the cancer risk and clinical prognosis of non-Hodgkin's lymphomas (NHL).Methods The single-nucleo-tide polymorphisms of rs3660 was genotyped with ligation detection reaction method.The association of rs3660 with NHL survival was calculated with log-rank test using Kaplan-Meier method.Multivariate survival analysis was performed using a Cox proportional hazards model.Results The rs3660 genotype distribution difference was not statistically significant between the case and control group (P =0.50).Patients carrying the rs3660 CG/CC genotype exhibited a significantly longer survival time than patients carrying the GG genotype (P =0.012).In addition,rs3660 was associated independently with the survival of NHL patients in multivariate analysis (RR=0.589,95％ CI:0.415-0.832,P =0.004).The association of this miR-SNP with NHL survival was further confirmed in the peripheral T cell lymphoma (PTCL) subtype.Conclusion Our results indicate that KRT81 rs3660 GG type is an independent prognostic marker in NHL.

The bovine enterovirus (BEV) is a pathogen found the digestive tracts of cattle. Recently, the BEV was discovered in cattle in a province in China. A rapid and effective detection method for the BEV is essential. An assay was carried out using two specific primers designed to amplify a highly conserved sequence of the 3D gene. A recombinant plasmid containing the target gene 3D was constructed as a standard control. The limit of detection of the reaction was 7.13 x 10(1) plasmid copies/μL of initial templates, which was tenfold more sensitive than the conventional reverse-transcription-polymerase chain reaction (RT-PCR). Moreover, the assay was highly specific because all negative controls and other viruses of clinical relevance did not develop positive results. Assay performance on field samples was evaluated on 44 (41 diarrhea and 3 aerosol) samples and compared with the conventional RT-PCR assay. Sixteen diarrhea samples were positive (16/41, 39. 02%) and 3 aerosol samples were positive (3/3, 100%). Preliminary results for clinical detection showed that the SYBR Green I real-time PCR assay was highly sensitive, specific and reproducible. The robustness and high-throughput performance of the developed assay make it a powerful tool in diagnostic applications for epidemics and in BEV research.
Animals ; Cattle ; Cattle Diseases ; diagnosis ; virology ; DNA Primers ; chemistry ; genetics ; Enterovirus Infections ; diagnosis ; veterinary ; virology ; Enterovirus, Bovine ; genetics ; isolation & purification ; Organic Chemicals ; chemistry ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

AIM:To investigate the effect of AG 490 on the expression of VEGF and HIF-1α, and the capacity of invasion in human erythroleukemia (HEL) cells.METHODS:The HEL cells were treated with AG490 at different con-centrations .The cell viability was detected by CCK-8 assay.The apoptosis was detected by Hoechst staining .The apoptosis and the cell cycle were analyzed by flow cytometry .The capacity of migration was evaluated by Transwell assay .The mRNA expression level of JAK2 was measured by RT-PCR.The protein levels of p-JAK2, VEGF and HIF-1αwere determined by Western blot.RESULTS:The HEL cell viabilities were 88%, 75%, 48%, 10%and 0.12%after treated with AG490 at 20, 40, 60, 80 and 100 μmol/L for 48 h, respectively.The results of Hoechst staining showed that brilliant blue cells in 80 μmol/L AG490 group was significantly increased compared with control group for 48 h.The apoptosis rate of 80μmol/L AG490 group was significantly increased compared with control group at 48 h after AG490 treatment.The number of membrane-permeating HEL cells in 20μmol/L AG490 group at 24 h after AG490 treatment was significantly lower than that in control group (P<0.05).The mRNA level of JAK2 decreased in a concentration-dependent manner after the HEL cells were treated with different concentrations of AG 490 for 48 h.The protein levels of p-JAK2, VEGF and HIF-1αwere lower in AG490 treatment groups than those in control group (P<0.05).CONCLUSION: AG490 inhibits the expression of VEGF and HIF-1αin HEL cells by inhibiting JAK2 pathway.

Objective To study the influence of 4-aminopyridine(4-AP)on proliferation,production,and apoptosis through inhibiting voltage-gated K+channel(Kv)in ovarian luteinized granulosa cells.nethods Ovarian luteinized granulosa cells were recovered from 25 women with regular menses who underwent in vitro fertilization programme.The cultured granulosa cells were divided into 4 groups:blank group,4-AP treated group,human chorionic gonadotropin(hCG)-induced group and hCG+4-AP cotreated group.The final concentrations of hCG and 4-AP were 1250 U/L and 5 nmol/L respectively.The progesterone production WaS detected by the chemoluminescence method.The expression of Kv mRNA on human ovarian luteinized granulosa cell was detected by RT.PCR The influence on the early apoptosis of gTanulosa cells bv 4-AP was observed by flow eytometry.Cellular caSpage-3 activities were observed with colorimetric method and the inhibition of the cell proliferation was studied using methyl thiazolyl tetrazolium(MTT)method.Results(1)Kv mRNA wag expressed in granulosa cell.(2)The progesterone production64),(206±32),(1991±172)and(763±79)nmol/L,respectively after24 hours culture.Exposure of the(3)The flow cytometry analysis and the cellular caapase-3 A405 showed that 4-AP increased the percentage ofearly phase apoptosis(P<0.01):4-AP treated group VS blank group[(40±5)%and 0.049 ±0.009]VS[(17±4)% and 0.029±0.008],hCG+4-AP CO-treated group VS hCG-induced group[(25±4)%and0.039 ±0.0081 VS[(15±3)%and 0.022 ±0.007].(4)24 hours after treated with 4-AP and hCG,theinhibitory rate of cultured granulosa cells of 4-AP treated group was higher than the blank group(19.7%VS0).and that of hCG+4-AP co-treated group was obviously higher than hCG-induced group(34.6% VS O,P<0.01).Conclusions The voltage-gated K+ channels expressed by ovarian luteinized granulosa cellplay an important role in cell proliferation,production,and apoptosis.4-AP may inhibit differentiation ofprogesterone in granulosa ceHs through the inhibition of proliferation and induction of apoptosis.

Objective To observe the prevalence of hyperuricemia(HUA)among obese children,and to ex-plore the association between uric acid(UA)levels and cardiometabolic risk factors,acanthosis nigricans and non - al-cohol fatty liver disease(NAFLD). Methods By using representative sampling method,1 753 obese children aged 6 -17 years old from 18 schools in 3 districts of Beijing(Xicheng,Haidian,Miyun)were selected to participate in the clini-cal examinations,including anthropometric measurements(height,weight)and blood pressure. Serum biochemical pa-rameters were assessed,including fasting plasma glucose(FPG),total cholesterol(TC),triglyceride(TG),high - densi-ty lipoprotein cholesterol(HDL - C),low - density lipoprotein cholesterol(LDL - C)and UA. Acanthosis nigricans and B - model ultrasonography of the liver were conducted. Results The prevalence of hypertension,impaired fasting glu-cose,dyslipidemia,acanthosis nigricans,and NAFLD among these 1 753 obese children was 33. 6%(589 cases), 66. 5%(1 156 cases),54. 3%(943 cases),23. 3%(408 cases),and 17. 0%(298 cases),respectively. The preva-lence of HUA was 40. 70%(714 / 1 753 cases),with 50. 17%(581 / 1 158 cases)in boys and 22. 34%(133 / 595 ca-ses)in girls. There was a significant increase in body mass index,systolic blood pressure,diastolic blood pressure, FPG,TG and LDL - C with the increase of UA,but there was a decrease in HLD - C with the increase of UA(all P ﹤0. 05). In boys,the adjusted odds ratios( OR)and 95% CI of the highest quartile of UA for hypertension,impaired fasting glucose,dyslipidemia,acanthosis nigricans,and NAFLD were 1. 16(0. 77 - 1. 74),1. 34(0. 90 - 1. 99),1. 29 (0. 89 - 1. 87),1. 89(1. 17 - 3. 04),and 1. 71(1. 03 - 2. 84),respectively;in girls,the adjusted OR and 95% CI of the highest quartile of UA for hypertension,impaired fas-ting glucose,dyslipidemia,acanthosis nigricans,and NAFLD was 0. 70(0. 40 - 1. 24),0. 60(0. 40 - 1. 00),1. 69(1. 04 - 2. 70),1. 67(0. 80 - 3. 49),and 1. 33(0. 48 - 3. 66),re-spectively. Conclusions The prevalence of HUA is relatively high in obese children and there is a strong association between UA and some car-diovascular metabolic disorders,acanthosis nigricans and NAFLD.

Objective To explore the effects of Chinese herbal compound Tangbikang on the expressions of p38 MAPK of sciatic nerve and plasma TNF-α in diabetic rats. Methods Ten of the sixty male SD rats were selected randomly as normal group, and the rest were fed with high-fat diet and low-dosage STZ was used to induce type Ⅱdiabetic rat models. Model rats were randomly divided into model group, mecobalamine group and Tangbikang low-, medium-, and high-dosage groups, 10 rats in each group. Each medication group was intervened with relevant medicine. Rat unilaterals sciatic nerves were taken after 16 weeks. The content of TNF-α in plasma was determined by radioimmunoassay. Western blot method was used to detect the expressions of p38 and p-p38 MAPK protein of sciatic nerve. Results Compared with normal group, the expressions of p38 and p-p38 protein and content of TNF-αin model group significantly increase (P<0.05, P<0.01). Compared with the model group, the expressions of p-p38 protein and the content of TNF-α significantly decreased after medicine intervention in different doses Tangbikang groups and mecobalamin group (P<0.05, P<0.01). The expression of p38 protein in Tangbikang high-dose group significantly decreased (P<0.05), with statistical significance (P<0.05). Conclusion Tangbikang can reduce the expression of p38 and p-p38 MAPK protein of the rat sciatic nerve, and reduce the content of TNF-α protein in rat plasma, which may be one of the effective targets of neuroprotection and abirritation of diabetic peripheral neuropathy.

Objective:To assess the predictive values of bioelectrical impedance analysis(BIA)-measured body fat indices to abnormal lipid profiles, and to preliminary propose optimal cut-off values of body fat in children and adolescents.Methods:Children and adolescents, aged 6-16 years, were selected from 30 schools (8 primary schools, 21 middle schools and one 12-year education school) in Dongcheng, Tongzhou, Fangshan and Miyun districts of Beijing by adopting a stratified cluster sampling method from November 2017 to January 2018.Questionnaire survey, body mass index(BMI), body fat mass index (FMI), fat mass percentage (FMP) and four lipid profiles were conducted.Results:A total of 14 309 participants, aged (11.0±3.3) years, were enrolled in the analysis, with 49.9% boys.In boys and girls, the percentile values ( P60- P95) fitted by FMI and FMP with K-median-coefficient of variation(LMS) method were taken as the cutting points, and P75 values were selected as the cut-off points of excessive body fat for their better sensitivity, specificity, predictive value and area under curve (AUC) for identification of abnormal lipid profiles.Boys with FMI above P75 accounted for 28% of the total population, and controlling boys with FMI below P75 could prevent dyslipidemia of 8%-57%.FMI in girl population occupied about 26% of the above, and controlling FMI in girl population below this cut-off point may prevent dyslipidemia from 8%-42%.FMP observed similar results to FMI.Assessed by FMI or FMP with P75 cut-off values, adiposity performed better than BMI for recognizing abnormal lipid profiles in boys (AUC: 52.4%-69.6% vs.50.2%-67.1%, P<0.05) rather than in girls ( P>0.05). In addition, when FMI or FMP beyond P90, the specificity of each abnormal lipid profiles was around 90%. Conclusions:The recommend cut-off points for body fat may be to assess children′s adiposity, and can be applied in preventive activities.

Paratuberculosis (Johne's disease) is a chronic debilitating disease of domestic and wild ruminants. However, widespread point-of-care testing is infrequent due to the lack of a robust method. The isothermal recombinase polymerase amplification (RPA) technique has applied for rapid diagnosis. Herein, RPA combined with a lateral flow dipstick (LFD) assay was developed to estimate DNA from Mycobacterium avium subsp. paratuberculosis. First, analytical specificity and sensitivity of the RPA-nfo primer and probe sets were assessed. The assay successfully detected M. paratuberculosis DNA in 30 min at 39℃ with a detection limit of up to eight copies per reaction, which was equivalent to that of the real-time quantitative polymerase chain reaction (qPCR) assay. The assay was specific, as it did not amplify genomes from five other Mycobacterium spp. or five pathogenic enteric bacteria. Six hundred-twelve clinical samples (320 fecal and 292 serum) were assessed by RPA-LFD, qPCR, and enzyme-linked immunosorbent assay, respectively. The RPA-LFD assay yielded 100% sensitivity, 97.63% specificity, and 98.44% concordance rate with the qPCR results. This is the first report utilizing an RPA-LFD assay to visualize and rapidly detect M. paratuberculosis. Our results show this assay should be a useful method for the diagnosis of paratuberculosis in resource-constrained settings.
Animals ; Diagnosis ; DNA ; Enterobacteriaceae ; Enzyme-Linked Immunosorbent Assay ; Genome ; Limit of Detection ; Methods ; Mycobacterium avium subsp. paratuberculosis ; Mycobacterium avium ; Mycobacterium ; Paratuberculosis ; Point-of-Care Testing ; Polymerase Chain Reaction ; Recombinases ; Ruminants ; Sensitivity and Specificity