Aim To explore the protective effect of PPARδ agonist GW501516 on neuro-vascular unit(NVU)injury induced by high glucose in vitro and its mechanism.Methods SD rat hippocampal neurons(Neu),astrocytes(Ast)and brain microvascular en-dothelial cells(BMEC)were isolated,purified and cultured in vitro,and NVU co-culture system was estab-lished.NVU co-culture system cells were divided into the control group,high glucose group(HG group),HG+GW501516 low,medium and high concentration groups(25,50 and 100 nmol·L-1)and HG+GW501516(100 nmol·L-1)+ANA12(TrkB inhibi-tor,5 μmol·L-1)group.NUV barrier function was e-valuated by transendothelial resistance(TEER)test and leakage test;the proliferative activity of Neu cells in co-culture system was detected by CCK-8 assay;the levels of inflammatory cytokines TNF-α,IL-6 and IL-1β in cell supernatant were detected by ELISA;the levels of SOD,MDA and NO in Neu cells were detected by chemical method;the apoptosis level was detected by flow cytometry;the protein expression levels of PPARδ,Bax,Bcl-2,cleaved caspase-3,and BDNF/TrkB pathway-related proteins BDNF,p-TrkB,and TrkB in Neu cells were detected by Western blot.Re-sults Compared with the control group,the TEER val-ue decreased and leakage value increased in HG group;the proliferation activity of Neu cells decreased,the levels of TNF-α,IL-6,IL-1β in supernatant and MDA and NO in Neu cells increased,and the SOD lev-el decreased;Neu cell apoptosis rate and expression levels of Bax and Cleaved caspase-3 increased,while the expression levels of PPARδ,Bcl-2,BDNF and p-TRKB/TrkB decreased(P＜0.05).Compared with the HG group,after treatment with different concentra-tions of GW501516,TEER value increased,leakage value decreased,proliferation activity of Neu cells in-creased,levels of TNF-α,IL-6,IL-1β in supernatant and MDA and NO in Neu cells decreased,and SOD level increased,and apoptosis rate of Neu cells and ex-pression levels of Bax and cleaved caspase-3 were de-creased,and expression levels of PPARδ,Bcl-2,BDNF and p-TRKB/TrkB increased(P＜0.05)in a dose-dependent manner.However,ANA12 intervention re-versed the effect of GW501516 on NVU damage under high glucose conditions.Conclusion PPARδ agonist GW501516 improves in vitro NVU injury induced by high glucose by activating BDNF/TrkB signaling path-way.

Objective:To explore the relationship between diffusion tensor imaging (DTI) parameter of fractional anisotropy (FA) and recovery of upper-extremity motor function in patients with ischemic stroke. Methods:From January to December, 2019, 20 ischemic stroke patients accepted routine medication and rehabilitation for three weeks. They received DTI examination and were measured FA of the infarct and the corresponding area on the contralateral side, the cerebral foot and the posterior limb of internal capsule of affected and unaffected sides, while the bilateral FA ratio (rFA) of them were calculated, before and after treatment. Meanwhile, all the patients were assessed with Fugl-Meyer Assessment-Upper Extremity (FMA-UE). Results:The FMA-UE score improved after treatment (t = 9.074, P < 0.001), while FA and rFA increased in infarct area (t > 14.519, P < 0.001). The difference before and after treatment of FA and rFA in all of the areas positively correlated with that of FMA-UE scores (r = 0.445~0.565, P < 0.05), which was the most in posterior limb of internal capsule of the affected side. Conclusion:FA of DTI alters with the recovery of motor function of the upper extremity after ischemic stroke, especially in the posterior limb of internal capsule.

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires

To compare the polysaccharides( LBLP),total flavonoids( LBLTF) and total phenols( LBLTP) of Lycium barbarum leaves from 14 production areas from Ningxia,Qinghai and Gansu at different harvesting time. Our results showed that there were significant difference of LBLP,LBLTF and LBLTP among different production areas. The LBLP collected from Xiangride township production area,Dulan,Qinghai was as high as 15. 02%. The LBLTF collected from Dagele township production area,Geermu,Qinghai was as high as28. 63%. The LBLTP collected from Keluke township production area,Delingha,Qinghai was as high as 16. 7%. There were also significant difference of these 3 components between different harvest periods. The average LBLP in May( 10. 20%) was significantly higher than that in June( 8. 49%). However,the average LBLTF( 17. 71%) and LBLTP( 12. 77%) in June was significantly higher than that in May( 14. 15%,11. 19%). The LBLTF and LBLTP have a positive correlation with the altitude of production areas,but there was no significant association between LBLP and altitude. The LBLP,LBLTF and LBLTP in different production areas and harvesting time was significant difference,the results provide some references for furthermore development and utilization of L. barbarum leaves.
Analysis of Variance ; China ; Flavonoids ; analysis ; Geography ; Lycium ; chemistry ; Phenols ; analysis ; Plant Leaves ; chemistry ; Polysaccharides ; analysis ; Seasons

OBJECTIVETo investigate the factors related to clinical pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET) in women with secondary infertility.

METHODSThe clinical, laboratory and follow-up data of 1129 cycles in 1099 patients with secondary infertility undergoing IVF-ET in Women's Hospital, Zhejiang University School of Medicine between July 2012 to July 2014 were retrospectively reviewed. The factors related to pregnancy outcomes were analyzed by univariate and logistic regression methods. The clinical pregnancy rates in women with different age and different number of embryos transferred were compared. The clinical outcomes of stimulation with gonadotropin releasing hormone (GnRH) agonist long protocol, GnRH agonist short protocol and GnH antagonist protocol were evaluated in secondary infertile patients aged ≥ 40 years.

RESULTSAmong 1129 cycles, 376 cases (33.30%) had clinical pregnancy and 753 cases (66.70%) had no clinical pregnancy. There were significant differences in age, body mass index, basal follicle-stimulating hormone level, antral follicle number,paternal age and number of embryos transferred between pregnancy and no pregnancy groups (P<0.05); while only maternal age (OR=0.900, 95% CI: 0.873~0.928, P<0.001) and the number of embryos transferred (OR=2.248, 95% CI: 1.906~2.652, P<0.001) were the independent factors affecting the clinical pregnancy outcome of IVF-ET. There was no significant difference in clinical pregnancy rate between women aged 30~40 years with two embryos transferred and those aged<30 years with two or three embryos transferred(P>0.05). There were no significances in clinical pregnancy rate among women aged ≥ 40 years using GnRH agonist long protocol,GnRH agonist short protocol and GnRH antagonist protocol for stimulation (P>0.05).

CONCLUSIONMaternal age and number of embryos transferred have independent effect on IVF-ET clinical pregnancy outcome of secondary infertile women. We suggest that no more than two embryos should be transferred for women in their thirties to minimize the risk of multiple pregnancy while still having an acceptable pregnancy rate. The pregnancy rate of patients over 40 years decreases significantly, and there is no difference in pregnancy rate by using GnRH agonist long protocol, GnRH agonist short protocol or GnRH antagonist protocol.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; Gonadotropin-Releasing Hormone ; agonists ; Gonadotropins ; Hormone Antagonists ; therapeutic use ; Humans ; Infertility, Female ; Maternal Age ; Ovarian Follicle ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies

Objective: To investigate the factors related to clinical pregnancy outcomes of in vitor fertilization-embryo transfer ( IVF-ET )in women with secondary infertility .Methods:The clinical , laboratory and follow-up data of 1129 cycles in 1099 patients with secondary infertility undergoing IVF-ET in Women’s Hospital, Zhejiang University School of Medicine between July 2012 to July 2014 were retrospectively reviewed.The factors related to pregnancy outcomes were analyzed by univariate and logistic regression methods .The clinical pregnancy rates in women with different age and different number of embryos transferred were compared .The clinical outcomes of stimulation with gonadotropin releasing hormone ( GnRH ) agonist long protocol , GnRH agonist short protocol and GnRH antagonist protocol were evaluated in secondary infertile patients aged≥40 years.Results: Among 1129 cycles, 376 cases( 333.0 %) had clinical pregnancy and 753 cases(66.70%)had no clinical pregnancy.There were significant differences in age , body mass index , basal follicle-stimulating hormone level , antral follicle number , paternal age and number of embryos transferred between pregnancy and no pregnancy groups(P<0.05);while only maternal age(OR=0.900, 95%CI:0 .873 ~0 .928 , P <0 .001 )and the number of embryos transferred (O R =2.248,95%CI:1.906~2.652,P<0.001)were the independent factors affecting the clinical pregnancy outcome of IVF-ET.There was no significant difference in clinical pregnancy rate between women aged 30 ~40 years with two embryos transferred and those aged<30 years with two or three embryos transferred ( P>0.05 ) .There were no significances in clinical pregnancy rate among women aged ≥40 years using GnRH agonist long protocol , GnRH agonist short protocol and GnRH antagonist protocol for stimulation(P>0.05).Conclusion: Maternal age and number of embryos transferred have independent effect on IVF-ET clinical pregnancy outcome of secondary infertile women.We suggest that no more than two embryos should be transferred for women in their thirties to minimize the risk of multiple pregnancy while still having an acceptable pregnancy rate .The pregnancy rate of patients over 40 years decreases significantly , and there is no difference in pregnancy rate by using GnRH agonist long protocol ,GnRH agonist short protocol or GnRH antagonist protocol .

Objective To study the efficacy of auxiliary early-adoption of Xinnao shutong capsules on anti-cerebral ischemia and neurological protection in senile patients with acute ischemic stroke. Methods Eighty senile patients with acute ischemic stroke were equally randomized into control group and treatment group. Conventional treatment was given to the control group; besides the conventional treatment, the Xinnao shutong capsules at a dosage of 3 pills and three- times daily for 30 d was given to the treatment group. The clinical efficacy and European stroke scale (ESS) scores were recorded and evaluated. Results The ESS scores in the treatment and control groups before the treatment were (56.24±10.48) and (57.73±9.85), respectively. The ESS scores in the treatment group on the 15th and 30th d of treatment (65.41±11.82, 72.52±13.21) was significantly improved as compared with those in the control group (59.52±10.13, 62.48±12.73) (P＜0.05). Significant differences on the clinical efficacy were noted between the 2 groups (Z=-2.813, P=0.005). Conclusion Xinnao shutong capsules have the effect of anti-cerebral ischemia and decreasing the neurological deficits in patients with acute ischemic stroke.

UNLABELLEDTo reduce the huge labor-cost in the screening in traditional monoclonal antibody generation, We established a new system for monoclonal antibody generation integrating with protein array. BALB/c mice were immunized by eight recombinant proteins respectively, and the positive hybridoma cells were obtained by cell fusion and ELISA screening. All the eight kinds of positive hybridoma cells were mixed, cloned, screened by protein array, and definite dilution cloned.

RESULTS175 single cell clones were obtained by complex cloning, and 119 of those were positive clones. Then 8 positive cell lines were generated by the following 2 rounds definite dilution cloning. By comparing with the traditional method, we got 8 monoclonal antibodies using the combined protein array screening and multiplex cloning method in 1 cycle, and fewer amounts of antigens were used. As a result, the combined protein array and multiplex cloning method could be used as an economical, rapid and simple tool applying in high throughput monoclonal antibody generation.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Female ; Humans ; Hybridomas ; metabolism ; Mice ; Mice, Inbred BALB C ; Protein Array Analysis