Objective:To investigate the effect of PD-1/PD-L1 inhibitor BMS-1 on LPS-induced inflammation in mouse microg-lia cells(BV-2 cells).Methods:Bv-2 cells were divided into Control group,LPS group and LPS+BMS-1 group.Bv-2 cells in Control group were cultured in DMEM medium for 78 hours,cells in LPS group were stimulated with 100 ng/ml LPS for 6 hours after 72 hours of normal culture,Bv-2 cells in LPS+BMS-1 group were treated with 50 nmol/ml BMS-1 for 72 hours and then stimulated with 100 ng/ml LPS for 6 hours.Expressions of PD-1 and iNOS mRNA in each group were detected by RT-qPCR,and expressions of PD-1 and iNOS protein in microglia were detected by Western blot.Flow cytometry was used to detect cell apoptosis in each group.Levels of inflamma-tory cytokines IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA.Results:RT-qPCR and Western blot results showed that com-pared with Control group,LPS group had significantly increased expression of PD-1 and iNOS(P<0.05).Compared with LPS group,LPS+BMS-1 group had significantly decreased expression of PD-1(P<0.05)and significantly increased expression of iNOS(P<0.05).Flow cytometry showed that compared with Control group,LPS group had a significantly increased in apoptosis of microglia(P<0.000 1).Compared with LPS group,LPS+BMS-1 group had a significantly increased in apoptosis of microglia(P<0.000 1).ELISA results showed that compared with Control group,LPS group had no significantly increased in pro-inflammatory factors IL-1β and IL-6(P>0.05),while significantly increased in TNF-α(P<0.000 1)and anti-inflammatory factor IL-10(P<0.000 1).Pro-inflammatory cyto-kine IL-1β in LPS+BMS-1 group was significantly higher than that in LPS group(P=0.000 1),IL-6 and TNF-α were also significantly higher than those in LPS group(P<0.000 1),while anti-inflammatory cytokine IL-10 in LPS+BMS-1 group was significantly lower than that in LPS group(P<0.000 1).Conclusion:BMS-1 can promote LPS-induced inflammatory response or impede the recovery of inflammation,and increase apoptosis of microglia.PD-1/PD-L1 pathway may be a potential therapeutic target for neuroinflammation.

Objective:To investigate the effect of PD-1/PD-L1 inhibitor BMS-1 on LPS-induced inflammation in mouse microg-lia cells(BV-2 cells).Methods:Bv-2 cells were divided into Control group,LPS group and LPS+BMS-1 group.Bv-2 cells in Control group were cultured in DMEM medium for 78 hours,cells in LPS group were stimulated with 100 ng/ml LPS for 6 hours after 72 hours of normal culture,Bv-2 cells in LPS+BMS-1 group were treated with 50 nmol/ml BMS-1 for 72 hours and then stimulated with 100 ng/ml LPS for 6 hours.Expressions of PD-1 and iNOS mRNA in each group were detected by RT-qPCR,and expressions of PD-1 and iNOS protein in microglia were detected by Western blot.Flow cytometry was used to detect cell apoptosis in each group.Levels of inflamma-tory cytokines IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA.Results:RT-qPCR and Western blot results showed that com-pared with Control group,LPS group had significantly increased expression of PD-1 and iNOS(P<0.05).Compared with LPS group,LPS+BMS-1 group had significantly decreased expression of PD-1(P<0.05)and significantly increased expression of iNOS(P<0.05).Flow cytometry showed that compared with Control group,LPS group had a significantly increased in apoptosis of microglia(P<0.000 1).Compared with LPS group,LPS+BMS-1 group had a significantly increased in apoptosis of microglia(P<0.000 1).ELISA results showed that compared with Control group,LPS group had no significantly increased in pro-inflammatory factors IL-1β and IL-6(P>0.05),while significantly increased in TNF-α(P<0.000 1)and anti-inflammatory factor IL-10(P<0.000 1).Pro-inflammatory cyto-kine IL-1β in LPS+BMS-1 group was significantly higher than that in LPS group(P=0.000 1),IL-6 and TNF-α were also significantly higher than those in LPS group(P<0.000 1),while anti-inflammatory cytokine IL-10 in LPS+BMS-1 group was significantly lower than that in LPS group(P<0.000 1).Conclusion:BMS-1 can promote LPS-induced inflammatory response or impede the recovery of inflammation,and increase apoptosis of microglia.PD-1/PD-L1 pathway may be a potential therapeutic target for neuroinflammation.

Objective To investigate the influence of serum cystatin C (Cys-C) and hypersensitivity C-reactive protein (hs-CRP) levels on the 3-year survival of patients undergoing maintenance hemodialysis (MHD). Methods A total of 358 patients with chronic renal failure who underwent MHD at the Second Affiliated Hospital of Nantong University from April 2011 to October 2020 were selected as study subjects. General clinical data, pre-dialysis laboratory test indicators, and echocardiographic indicators 3 months after dialysis were recorded. The survival status of patients after 3 years of dialysis was followed up, and the general clinical data, pre-dialysis laboratory test indicators, and echocardiographic indicators 3 months after dialysis were compared between surviving and deceased patients. Univariate and multivariate Cox regression analyses were performed to screen influencing factors of 3-year survival in MHD patients. Results At the 3-year follow-up, of the 302 MHD patients' 203 survived, and 99 died. Statistically significant differences were observed in age, primary disease, diabetes status, congestive heart failure, statin use, antiplatelet drug use, diuretic use, dialysis mode, estimated glomerular filtration rate (eGFR) and gamma-glutamyl transferase (GGT), alkaline phosphatase (AKP), total bilirubin (TBIL), β₂-microglobulin (β₂-MG), creatinine (Cr), low-density lipoprotein cholesterol (LDL-C), hypersensitive C-reactive protein (hs-CRP), and serum phosphorus (P) levels between surviving patients and deaths(P < 0.05 or P < 0.01). Univariate Cox regression results showed that age of MHD patients, primary disease (diabetic nephropathy, hypertensive nephropathy, polycystic kidney disease, others), comorbidities(diabetes, congestive heart failure, other cardiovascular diseases), drug use (statins, antiplatelet drugs), dialysis method (hemodialysis, hemodialysis + perfusion), laboratory test indicators [GGT, AKP, TBIL, total bile acid (TBA), albumin (ALB), Cr, Cys-C, eGFR, apolipoprotein A (APO-A), hs-CRP]were all influential factors of 3-year survival rate of MHD patients. Multivariate Cox regression analysis revealed that age, primary disease, other cardiovascular diseases, dialysis mode, AKP, ALB, TBIL, Cys-C, and hs-CRP were significant influencing factors for the survival of patients with chronic renal failure (P < 0.05). Conclusion Serum Cys-C and hs-CRP levels before MHD in patients with chronic renal failure may be associated with their 3-year survival after dialysis treatment. High serum Cys-C may reduce the risk of poor clinical prognosis, while high serum hs-CRP may increase the risk of poor clinical prognosis.

Objective:To investigate the distributions of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) genotypes among newly reported HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province from 2016 to 2019.Methods:A total of 1 289 newly reported HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture were collected through the National Acquired Immunodeficiency Syndrome Comprehensive Prevention and Control Data Information System From January 2016 to December 2019. Among them, 996 subjects with a plasma volume of ≥200 μL were selected to perform HIV and HCV genotyping. The HIV pol gene, the HCV core protein-binding envelope protein ( CE1) gene and non-structural protein 5B ( NS5 B) gene were amplified using the nested polymerase chain reaction.The phylogenetic tree was constructed by MEGA 7.0 software to classify the genotypes. Chi-square test was used for statistical analysis. Trend chi-square test was used to analyze the trend of HIV and HCV genotypes. Results:Among the 996 cases with HIV/HCV co-infection, HIV and HCV sequences from a total of 554 subjects (55.6%, 554/996) were successfully obtained, and the genotypes of HIV and HCV were diverse. HIV genotype C (40.3%, 223/554) and BC recombinant (33.6%, 186/554) were the most prevalent, followed by genotype B (6.5%, 36/554) and circulating recombinant form (CRF)01_AE (3.6%, 20/554). HCV genotype 3b was the most prevalent (31.2%, 173/554), followed by genotype 6u (19.5%, 108/554), 1a (17.5%, 97/554), 6n (11.4%, 63/554), 3a (8.7%, 48/554) and 6xg (6.3%, 35/554). The prevalence of HIV genotype C showed a downward trend ( χtrend2=7.23, P<0.001), while the prevalence of BC recombinant showed an upward trend ( χtrend2=5.97, P<0.001), and the proportion of BC recombinant was higher than genotype C in 2019 (54.9%(101/184) vs 21.7%(40/184)). However, there were no statistically significant differences in the proportions of genotype 3b, 6u and 1a from 2016 to 2019 ( χtrend2=1.43, 1.79 and 0.39, respectively, P=0.152, 0.074 and 0.695, respectively). The HIV genotype distribution among patients with different ethnic groups were significantly different ( χ2=22.06, P=0.037). Conclusions:The diversity of HIV and HCV genotypes is high and complex among HIV/HCV co-infected Burmese patients in Dehong Dai and Jingpo Autonomous Prefecture. BC recombinant shows a trend of becoming the predominant HIV genotype among these co-infected patients. Therefore, surveillance of the prevalence of HCV and HIV genotypes in Burmese population needs to be further strengthened.

Objective:To search and summarize the best evidence for blood pressure management in patients with intracerebral hemorrhage to provide evidence for blood pressure management in these patients.Methods:PIPOST model was used to establish evidence-based questions; with "intracerebral hemorrhage/cerebral hemorrhage/ICH/HICH/stroke" AND "blood pressure/hypertension" as English search terms, and with "脑出血/颅内出血/脑卒中"+"血压" as Chinese search terms, the official websites of related associations and domestic and foreign databases, such as GUIDE.com, UpToDate, Cochrane Library, CNKI, VIP Chinese journal Service platform, and Wanfang Data Knowledge Service platform, were searched. Two researchers independently evaluated the quality of the literature, including guidelines, systematic reviews, and expert consensus, and extracted the evidence.Results:A total of 17 articles were selected; 2 were clinical decision type, 7 were guideline type, 7 were systematic review type, and 1 was expert consensus type. Twenty-six pieces of evidence were summarized into 5 aspects: timing and methods of lowering blood pressure, benefits and risks of lowering blood pressure, drug selection and selection methods, recurrence prevention of stroke, and matters for attention.Conclusion:It is necessary to pay attention to blood pressure management in patients with intracerebral hemorrhage; in the process of evidence application, it's suggested to make an appropriate blood pressure management plan for the patients according to clinical situation.

Objective:To comparatively analyze for the fundus characteristics of acquired immune deficiency syndrome (AIDS) with cytomegalovirus retinitis (CMVR) between first-visit and non-first-visit in ophthalmology.Methods:A retrospective study was performed for 22 patients (41 eyes) diagnosed as CMVR with AIDS by ophthalmology in the Affiliated Municipal Hospital of Xuzhou Medical University from July 2004 to September 2017. The patients were divided into two groups: one with the first-visit in ophthalmology (FVO) and the other with the first-visit in non-ophthalmology (FVNO). All patients underwent visual acuity, intraocular pressure, slit lamp microscope and indirect ophthalmoscope examinations. Thirty-nine eyes of 21 patients with clear refractive media were examined by color fundus photography, of which 5 patients were examined by FFA. Five patients examined by OCT and B-mode ultrasound. The CMVR were characterized as fulminant type or indolent type. All the 22 patients (41 eyes) except 2 patients (4 eyes) accepted highly active antiroviral therapy, and all patients were treated with ganciclovir intravenously. Nine patients (12 eyes) received intravitreal injection of ganciclovir, 7 patients (10 eyes) underwent vitrectomy because of retinal detachment, 6 patients (7 eyes) gave up surgery because of extensive retinal detachment, and the other 12 eyes did not undergo any eye surgery. All patients were followed for 6-58 months (average 23±39 months). The incidences and fundus characteristics of the patients with fulminant or indolent CMVR were compared and analyzed. Between the two groups, the difference of visual acuity and CD4 +T cell count at the first vist and the last follow-up were analyzed by a t-test, and the macular involvement and spread of the two groups were compared by a chi-square test. Results:Fulminant CMVR showed dense yellow-white necrotic lesions along the great vessels with or without satellite lesions, while indolent CMVR showed sparse yellow-white granular lesions with a little bleeding. The concomitant signs included frost-like dendritic vascular sheath, retinal artery occlusion and optic disc edema. Fourteen eyes of 7 patients in FVO group were fulminant, including 2 patients (2 eyes) with retinal artery occlusion and 1 patient (two eyes) with optic disc edema. In FVNO group, there were 27 eyes of 15 patients, including 21 eyes of 13 cases identified as fulminant type and 6 eyes of 4 patients as indolent type. In the fulminant type, there were 4 patients (6 eyes) with frost-like dendritic vascular sheath, 10 patients (12 eyes) with retinal artery occlusion and 4 patients (4 eyes) with optic disc edema. There was no significant difference in initial visual acuity ( t=-1.534, P=0.133), but there was a significant difference in visual acuity at the last follow-up ( t=-3.420, P=0.001). There was no significant difference in CD4 + T cells between the two groups at the first visit ( t=-0.902, P=0.378). The proportions of macular involvement and 3-4 quadrant involvement in FVO group were significantly higher than those in FVNO group ( χ 2=7.552, 7.865; P=0.006, 0.005). Conclusion:For AIDS patients with CMVR, the first-visit in ophthalmology showed more dense necrotic lesions involving macular and a wider range of lesions than the first-visit in non-ophthalmology.

Objective To investigate the effects of NANOG/deleted in breast cancer 1 (DBC1)pathway on biological behavior of gastric cancer cells.Methods From May 2014 to May 2015,25 patients who underwent gastric cancer resection were selected.The expression of NANOG and DBC1 was detected by real time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting in N-tera,SGC-7901,HGC-27,MKN-45,MGC803,NCI-N87,BGC823 cell lines,normal gastric epithelial cell line GES-1 and gastric cancer tissues.The proliferation,apoptosis and colony formation ability of MKN-45 cells in short hairpin (sh)NANOG-1,shNANOG-2,sh-control and shDBC1 groups were determined by MTT assay,flow cytometry and colony forming assay.The effects on the expression of the two genes in MKN-45 cells were verified by polymerase chain reaction (PCR) in shNANOG,sh-control,shDBC1 and shDBC1+NANOG groups and the effects of down regulation of DBC1 on cell biological behavior were further investigated.The differences in gene expression profile after interference which were screened by gene chips and bioinformatics were analyzed.The mechanism of NANOG regulating DBC1 was explored by Dual-luciferase assay.T test was used for two groups comparison while one-way analysis of variance was for multiple groups.Results NANOG and NANOG mRNA were highly expressed in N-tera cells,which were 1.02±0.08 and 0.95 ±0.03,respectively,and the expressions in SGC-7901,HGC-27,MKN-45 and NCI-N87 cell lines were 0.67±0.03 and 0.64±0.04,0.58±0.02 and 0.28±0.02,0.83±0.03 and 1.04 ± 0.05,and 0.61 ± 0.02 and 0.64 ± 0.08,respectively;no expression was detected in normal gastric epithelial cell line GES-1,and the expressions in MKN-45 cells were the highest in gastric cancer cells (F=21.51 and 85.53,both P＜0.01).The expression of DBC1 in HGC-27,MGC803,NCIN87,SGC-7901,BGC823 and MKN-45 cells were 0.37±0.02,0.33±0.02,0.42±0.01,0.58±0.04,0.33±0.05,and 0.87±0.02,respectively;while there was no expression of NANOG,NANOG mRNA and DBC1 in GES-1 cells.The expression of NANOG mRNA and DBC1 was detected in gastric cancer tissues of 24.0％ (6/25) patients.Compared with that of the sh control group,the apoptosis rates of MKN-45 cells in the shNANOG-1,shNANOG-2 groups were increased ((2.24±0.17)％ vs (6.03±0.24) ％ and (6.95 ± 0.38) ％),and the difference was statistically significant (F =81.18,P ＜ 0.01).Compared with that of the sh-eontrol group,the colony forming abilities of MKN-45 cells in the shNANOG-1 and shNANOG-2 groups were significantly decreased (172.03±6.35 vs 74.32±5.32 and 53.08±3.82),and the difference was statistically significant(F=171.61,P＜0.01).The results of PCR showed that compared with that of sh-eontrol group,the expression levels of NANOG mRNA and DBC1 mRNA in shNANOG group were lower (1.04±0.05 vs 0.54±0.03,1.08±0.08 vs 0.42±0.03),the level of DBC1 mRNA in shDBC1 group was lower (1.08±0.08 vs 0.50±0.04),and the differences were statistically significant (t=9.15,7.37 and 6.06,all P＜0.01).The expression level of NANOG mRNA in shDBC1 + NANOG group was higher (1.04 ± 0.05 vs 3.01 ± 0.08),while the expression level of DBC1 mRNA was lower (1.08 ± 0.08 vs 0.71 ± 0.06),and the difference was statistically significant (t=-20.22 and 3.74,both P＜0.05).The expression level of DBC1 mRNA in shDBC1±NANOG group was higher than that in shDBC1 group (0.71±0.06 vs 0.50±0.04),and the difference was statistically significant (t=4.00,P＜0.05).Bioinformatic analysis showed that DBC1 gene promoter region had the potential NANOG protein binding sites.Dual-luciferase assay indicated NANOG played the role in transcription activation in DBC1 promoter regions.Conclusion NANOG and DBC1 are highly expressed in various gastric cancer cell lines.NANOG may affect the proliferation,apoptosis and colony formation of MKN-45 cells by regulating the expression of DBC1.NANOG/DBC1 pathway may be a promising new target of gastric cancer treatment.

In order to understand the existing problems in the management of medical ethical review,this paper summed up related domestic literature published in 2001 to 2015 with the method of literature analysis.There existed several problems in the construction and management,the ability of ethical review,external supervision,and other aspects of the current medical ethics committee.It suggests that strengthening external continuous supervision,strengthening the ethics committee itself construction and management and participating in standardized certification can help medical ethics committee to solve the above problems and have important significance for the standardized development.

Objective To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the generation of human myeloid derived suppressor cells (MDSCs) relied on peripheral blood monocytes,and to establish efficient induction system in vitro of MDSCs.Methods Kidney transplantation recipients between January and March 2017 were included in this study.Purified CD14 + cells isolated from peripheral blood were cultured in the presence of GM-CSF with different concentrations for 7 days.Phenotypes and immunosuppressive abilities of induced MDSCs (iMDSCs) were investigated with FACS analyses.Inducible nitric oxide synthase (iNOS) mRNA expression was detected by qRT-PCR to determine the influence of iNOS-pathway on the immunosuppressive abilities of iMDSCs.Results A total of 11 recipients were included in this study.HLA-DR expression decreased sharply after the culture with GM-CSF.iMDSCs showed the similar phenotype characteristics with monocytic-MDSCs (M-MDSCs) as well as significant ability to suppress T cells proliferation and cytokines production.iMDSCs expressed higher levels of iNOS than monocytes,and the inhibitor effects of iMDSCs were significantly reduced after treatment with L-NMMA (1 mmol/L).The variations of phenotype and suppressive ability were concentrationdependent,and more significant changes could be revealed in the group of 10 μg/L GM-CSF.Conclusion GM-CSF-treated peripheral blood monocytes can be efficiently transformed to M-MDSCs,and suppress T cells proliferation and cytokines secretion via iNOS-dependent pathway.These results may contribute to establish MDSCs induction system,which would provide a basis for the clinical application of MDSCs.

Objective To investigate the effects of commonly used inductive agents on peripheral blood monocytic myeloid-derived suppressor cells (M-MDSCs) in renal transplantation recipients and to discuss their possible mechanism.Methods The enrolled patients received rabbit anti-thymocyte globulin (rATG) or basiliximab for induction therapy,with the maintenance immunosuppressive regimen of tacrolimus,mycophenolate mofetil and steroid.The number of CD11 b + CD33 + HLA-DR-CD14 + CD1 5-M-MDSCs and cytokine levels in peripheral blood,including interferon-γ(IFN-γ),interleukin-2 (IL-2),IL-4 and IL-6,were measured by flow cytometry before and 1 week,2 weeks,1 month,2 months,3 months after operation.Results A total of 47 recipients (29 given rATG 29,and 18 given basiliximab) were included in this study.Compared to the patients with basiliximab,asignificant increase in the frequency of M-MDSCs was observed in the rATG group at 2nd month after operation (5.5％ ± 2.8％ vs.3.8％ ± 1.6％,P＜0.001) and at 3rd month after operation (7.0 ％ ± 3.1％vs.4.1％ ± 2.3 ％,P＜ 0.001),while there was no significant difference in the cell number between the two groups.In the cytokine detection,levels of IL-2 and IL-4 in the rATG-treated recipients were significantly higher at 2nd weekpostoperation (Pr2 =0.032,and PIL-4 =0.019)and 1st month postoperation (PIL-2 =0.024,PIL-4 ＜0.001) than the basiliximab group.Conclusions ATG promotes the expansion of M-MDSCs,which is associated with the secretion of IL-2 and IL-4 due to the lymphocytes depletion.The synergistic immunosuppressive effect may contribute to the induction of immune tolerance.