Objective:To obtain a high specificity and high affinity anti-human PD-L1 monoclonal antibody which can be used for clinical diagnosis and block PD-L1 and PD-1 binding.Methods:BALB/c mice were immunized with recombinant PD-L1 protein.The positive cell clones stably secreting anti-human PD-L1 monoclonal antibody were obtained by classical hybridoma cell fusion technique.The specificity,affinity,subtype and other characteristics of the antibody were identified by ELISA.Immunofluorescence and indirect immunofluorescence were used to detect the tumor cells.Antibody blocking activity was confirmed by tumor killing test.Results:Two cell strains stably secreting monoclonal antibodies against human PD-LI were screened out.Abl and Ab2 had high titer and affinity.The antibody titers were 1:2.56×106 and 1:3×105,and the affinity was 1.5×109 L/mol and 2.5×10s L/mol respectively.There was no cross reaction between these two antibodies and PD-L2.Immunoblotting,indirect immunofluorescence confirmed that the antibody can be used to the diagnosis.Experiment showed that PD-L1 antibodies can increases tumor-killing activity of CIK cells.Conclusion:Two hybridoma cell lines capable of stably secreting highly specific and high affinity anti-human PD-L1 monoclonal antibody are obtained.They can specifically bind to PD-L1 molecules on tumor cells and can be used to the diagnosis of tumor phenotype and prognosis.Antibody blocking function can be applied to combined CIK cell immunotherapy.

Objective To investigate the effect of moderate treadmill exercise together with modified hydroxyapatite chitosan composite hydrogel (CS/HA-g-CS) implantation on repair of full-thickness defects of articular cartilage in rats.Methods Full-thickness cartilage defects were drilled in the patellar groove of bilateral femoral condyles in a total of 24 male SD rats before they were randomly assigned into 4 groups.The control group (BC group) was subjected to no exercise or CS/HA-g-CS implantation;the chitosan group (CHI group) to CS/HA-g-CS implantation without exercise;the moderate treadmill exercise group (MIR group) to exercise 4 weeks after modeling without CS/HA-g-CS implantation;the CHI + MIR group to moderate treadmill exercise plus CS/HA-g-CS implantation 4 weeks after modeling.Half of the animals were sacrificed at week 8 and half at week 16 after operation.Femoral condyles were harvested for gross observation and histochemical measurement by O' Driscoll scoring system.mRNA expressions of glycosaminoglycan,collagen type Ⅱ and BMP-2 were detected by RT-PCR.Results Gross observation revealed:at 8 weeks after modeling,the CHI + MIR group was significantly better than the other 3 groups,with the BC group in the poorest (P ＜ 0.05);at 16 weeks after modeling,the BC group was significantly poorer than the other 3 groups (P ＜0.05) among which there were no significant differences (P ＞ 0.05).O 'Driscoll scoring revealed:at both 8 and 16 weeks after modeling,the CHI + MIR group was significantly better than the other 2 groups and the BC group significantly poorer than the other 3 groups (P ＜ 0.05) but there was no significant difference between the MIR and CHI + MIR groups(P ＞ 0.05).The expressions of collagen type Ⅱ,glycosaminoglycan and BMP-2 were significantly higher in the CHI + MIR group than in the other 3 groups (P ＜ 0.05) and significantly lower in the BC group than in the other 3 groups (P ＜ 0.05).Conclusions Moderate treadmill exercise together with CS/HA-g-CS implantation has significant positive effects on repair of full-thickness defects of articular cartilage in rats than merely moderate treadmill exercise or CS/HA-g-CS implantation alone.The defective cartilage repaired by moderate treadmill exercise together with CS/HA-g-CS implantation contains more collagen type Ⅱ and glycosaminoglycan and shows morphology of nearly normal cartilage.

OBJECTIVE:To evaluate the efficacy and safety of cerebrolysin in adjuvant treatmenut of acute cerebral infarction systematically,and to provide evidence-based reference in clinic.METHODS:Retrieved from SCI,Cochrane Library,EMBase,PubMed,CJFD,VIP and Wanfang Database,RCTs about cerebrolysin combined with routine plan (trial group) vs.routine plan alone or combined with placebo (control group) in adjuvant treatment of acute cerebral infarction were collected.After data extraction and quality evaluation by using Cochrane systematic review manual 5.1.0,Meta-analysis was conducted by using Rev Man 5.2 statistical software.RESULTS:A total of 20 RCTs were included,involving 3 313 patients.Meta-analysis showed that NIHSS score [MD=-1.77,95％CI(-2.33,-1.21),P＜0.001],response rate [OR=2.85,95％CI(1.75,4.63),P＜0.001] and Barthel index (BI) score [MD =7.30,95 ％ CI (3.48,11.13),P＜ 0.001] in trial group were significantly higher than control group,with statistical significance.There was no statistical significance in disability rate [OR=0.46,95％ CI(0.20,1.03),P=0.06],mortality [OR=0.79,95％ CI (0.52,1.19),P=0.25],the incidence of ADR [OR=1.04,95％ CI (0.85,1.27),P=0.72] or the incidence of severe ADR [OR=0.01,95％CI(-0.02,0.04),P=0.51] between 2 groups.CONCLUSIONS:Cerebrolysin is good for adjanctive therapy of acute cerebral infarction,can significantly improve neurologic impairment and life quality and dosen't increase the incidence of ADR.

Objective To investigate the prevalence of restless legs syndrome (RLS) in peritoneal dialysis patients and analyze the related risk factors.Methods This study was a cross-sectional study.The patients receiving maintenance peritoneal dialysis from January 2017 to December 2017 in the Peritoneal Dialysis Center of the Second Hospital Affiliated to Soochow University were selected as the study subjects.RLS was screened for peritoneal dialysis patients by epidemiological field investigation based on the RLS diagnostic criteria of the International Restless Leg Syndrome Research Group in 2014.Clinical data and laboratory examinations of selected patients were collected and the differences of clinical indicators between RLS and non-RLS patients were compared.The risk factors related to RLS were analyzed by logistic regression.Results Seventy-six cases of RLS were screened out from 396 PD patients.The prevalence of RLS was 19.2％.Compared with non-RLS group,RLS group patients had longer dialysis age,less 24 hours urine volume,and elevated blood intact Parathormone (iPTH) and alkaline phosphatase (AKP) (all P ＜ 0.05).There was no significant difference in primary disease ratio,sex,age,body mass index,blood pressure,hemoglobin,creatinine,urea nitrogen,uric acid,ferritin,serum iron,transferrin saturation,blood calcium,blood phosphorus,total cholesterol,triglyceride,low density lipoprotein,high density lipoprotein,eGFR,Kt/V,Ccr between RLS and non-RLS group patients (all P ＞ 0.05).Multivariate logistic regression analysis showed that long dialysis age (OR=1.010,95％CI 1.001-1.018,P=0.022) and high blood AKP (OR=1.005,95％CI 1.001-1.010,P=0.021) were independent risk factors for RLS in peritoneal dialysis patients (both P ＜ 0.05).Conclusions The prevalence of RLS is high in peritoneal dialysis patients.Long dialysis age and high blood AKP are independent risk factors for RLS.

Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults and is poorly controlled. Previous studies have shown that both macrophages and angiogenesis play significant roles in GBM progression, and co-targeting of CSF1R and VEGFR is likely to be an effective strategy for GBM treatment. Therefore, this study developed a novel and selective inhibitor of CSF1R and VEGFR, SYHA1813, possessing potent antitumor activity against GBM. SYHA1813 inhibited VEGFR and CSF1R kinase activities with high potency and selectivity and thus blocked the cell viability of HUVECs and macrophages and exhibited anti-angiogenetic effects both in vitro and in vivo. SYHA1813 also displayed potent in vivo antitumor activity against GBM in immune-competent and immune-deficient mouse models, including temozolomide (TMZ) insensitive tumors. Notably, SYHA1813 could penetrate the blood-brain barrier (BBB) and prolong the survival time of mice bearing intracranial GBM xenografts. Moreover, SYHA1813 treatment resulted in a synergistic antitumor efficacy in combination with the PD-1 antibody. As a clinical proof of concept, SYHA1813 achieved confirmed responses in patients with recurrent GBM in an ongoing first-in-human phase I trial. The data of this study support the rationale for an ongoing phase I clinical study (ChiCTR2100045380).