Background::Programmed death 1 (PD-1) blockade plus chemotherapy has become the new first-line standard of care for patients with advanced non-small-cell lung cancer (NSCLC). Yet not all NSCLC patients benefit from this regimen. This study aimed to investigate the predictors of PD-1 blockade plus chemotherapy in untreated advanced NSCLC.Methods::We integrated clinical, genomic, and survival data from 287 patients with untreated advanced NSCLC who were enrolled in one of five registered phase 3 trials and received PD-1 blockade plus chemotherapy or chemotherapy alone. We randomly assigned these patients into a discovery cohort ( n = 125), a validation cohort ( n = 82), and a control cohort ( n = 80). The candidate genes that could predict the response to PD-1 blockade plus chemotherapy were identified using data from the discovery cohort and their predictive values were then evaluated in the three cohorts. Immune deconvolution was conducted using transcriptome data of 1014 NSCLC patients from The Cancer Genome Atlas dataset. Results::A genomic variation signature, in which one or more of the 15 candidate genes were altered, was correlated with significantly inferior response rates and survival outcomes in patients treated with first-line PD-1 blockade plus chemotherapy in both discovery and validation cohorts. Its predictive value held in multivariate analyses when adjusted for baseline parameters, programmed cell death ligand 1 (PD-L1) expression level, and tumor mutation burden. Moreover, applying both the 15-gene panel and PD-L1 expression level produced better performance than either alone in predicting benefit from this treatment combination. Immune landscape analyses revealed that tumors with one or more variation in the 15-gene panel were associated with few immune infiltrates, indicating an immune-desert tumor microenvironment.Conclusion::These findings indicate that a 15-gene panel can serve as a negative prediction biomarker for first-line PD-1 blockade plus chemotherapy in patients with advanced NSCLC.

Objective:To investigate the correlation of the expression of Ephrin A receptor 2(EphA2)and its promoter region DNA hypomethylation with the occurrence of pyroptosis in invasive breast cancer. Methods:The expression level of pyroptosis-related protein EphA2 in normal breast tissue,paracancerous tissues and cancer tissues from 42 breast cancer patients was examined by Immunofluorescence staining and Western blotting.The expression level of pyroptosis related protein nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)was analyzed by immunofluorescence staining and Western blotting.The expression levels of apoptosis related proteins Caspase 1 and inflammatory cytokine interleukin-1β(IL-1 β)were studied by Western blotting.The DNA methylation level in the promoter region of EphA2 was investigated by nested methylation-specific PCR(nMS-PCR).The expression levels of DNA methyltransferase 1(DNMT1)and DNA methyltransferase 3a(DNMT3a)were examined by Western blotting.The correlation of the protein expression and methylation level of EphA2 in cancer tissues with the expression NLRP3,Caspase 1,IL-1 β,DNMT1 and DNMT3a was analyzed by Pearson correlation coefficient. Results:Compared with normal breast tissues and paracancerous tissues,the expression level of EphA2 protein was significantly increased(P＜0.01),while that of NLRP3,Caspasel and IL-1 βwas significantly decreased(P＜0.05)in breast cancer tissues.Meanwhile,compared with normal breast tissues and paracancerous tissues,the DNA methylation level of EphA2 promoter in breast cancer tissues was significantly decreased(P＜0.05),the expression level DNMT3a protein was significantly decreased(P＜0.01,P＜0.05),and the difference in the expression level of DNMT1 protein was not statistically significant.Pearson correlation analysis showed that the expression level of EphA2 protein is negatively correlated with that of NLRP3(r=-0.651 2,P＜0.05),Caspasel(r=-0.571 2,P＜0.05),IL-1β(r=-0.654 6,P＜0.05)or DNMT3a(r=-0.537 4,P＜0.05),while the methylation level of EphA2 was positively correlated with the protein expression level of NLRP3(r=0.634 1,P=0.026 8),Caspase1(r=0.672 8,P=0.01 6 5),IL-1 β(r=0.694 0,P=0.01 2 3)and DNMT3a(r=0.687 1,P=0.01 3 6). Conclusion:The expression of EphA2 protein is upregulated in breast cancer tissues is negatively correlated with pyroptosis.DNMT3a may be involved in the process of DNA hypomethylation in the promoter region of EphA2.

Objective:To identify the transmission relationship between HIV infection cases the non-marital non-commercial heterosexual contact in Zhejiang Province.Methods:When HIV positive was informed during January 2020 to January 2022, the staff conducted an epidemiological investigation to collect cases information on sociodemographic characteristics, mobility information, past HIV testing history, high-risk sexual behaviors, sexual partners, and etcetera. At the same time, 6-8 ml of blood from the new diagnosis of people infected with HIV before antiviral treatment was collected to separate the bleeding plasma. pol gene was amplified by nucleic acid extraction and PCR, sequenced by Sequencer 5.0 software, and Cytoscape 3.6.0 software was used to draw HIV molecular transmission network. Results:From January 2020 to January 2022, 88 HIV infected individuals were found in Pujiang County, of which 74 were transmitted through heterosexual transmission, of which 31 were infected through non-marital non-commercial heterosexual contact. Preliminary case studies have found that three female cases have engaged in unprotected non-marital non-commercial heterosexual contact with one male case. Among the 4 infected individuals, 2 of their spouses tested positive for HIV antibodies. Molecular transmission network monitoring was carried out on 65 newly diagnosed cases of heterosexual transmission with acquired sequences, forming 9 transmission clusters. The largest cluster contained 10 cases. A total of 11 HIV-infected individuals were involved in this HIV cluster epidemic. They were 3 males and 8 females, all over 50 years old and were farmers or rural housewives. They were traced to 7 sexual partners (6 negatives of HIV, 1 undetected). Among the 18 respondents' sexual social network relationships, there were 6 couples, 8 permanent partners, and 3 temporary partners. Among 11 HIV infected individuals, there were 9 cases of non-marital non-commercial heterosexual transmission and 2 cases of intramarital transmission. The epidemiological association between 7 non-married non-commercial heterosexual partners and case 2 (56-year-old male farmer), 3 cases confirmed by epidemiological investigation and molecular transmission cluster results, 3 cases confirmed by molecular transmission cluster and epidemiological investigation results, and 1 case confirmed by epidemiological investigation results.Conclusions:The transmission mode of this cluster epidemic was to spread HIV through heterosexual sex with a male case as the core, then cause the transmission within marriage and between fixed sexual partners. The combination of epidemiological investigation and molecular transmission network traceability survey supports the conclusion of this study.

Objective: To analyze the characteristics and influencing factors of the newly diagnosed cases locally infected with HIV through sexual contact in Jinhua,so as to provide reference for AIDS prevention and control. Methods: An epidemiological survey was conducted among the HIV/AIDS cases diagnosed in 2017 in Jinhua to collect the information about demographic characteristics,local infection and sexual behaviors. The logistic regression model was used to analyze the related factors for local HIV infection. Results: A total of 438 HIV/AIDS cases infected through sexual contact were recruited,with 272（62.10%）cases infected in Jinhua. The proportion of local infection was 86.67%,79.47%,63.04%,69.09%,77.46% and 77.97%,respectively,among those people aged 60 years or over,permanent residents in Jinhua,employees / students,farmers,those who had lived in Jinhua for more than five years,and those who had a HIV testing in one year. The Results of the multivariate logistic regression analysis showed that heterosexual transmission cases who were permanent residents in Jinhua（OR=3.437,95%CI：1.250-9.451）,who had lived in Jinhua for more than five years（OR=3.609,95%CI：1.403-9.284）,who had commercial heterosexual behaviors in Jinhua（OR=5.463,95%CI：2.529-11.803）were more likely to be infected with HIV in Jinhua;homosexual transmission cases who were permanent residents in Jinhua（OR=4.812,95%CI：1.744-13.275）and who had non-commercial,temporary homosexual behaviors in Jinhua（OR=10.641,95%CI：4.369-25.916）were more likely to be infected with HIV in Jinhua. Conclusion Among the HIV/AIDS cases diagnosed in 2017 infected through sexual contact in Jinhua,having permanent residence,long-term residence,commercial heterosexual behaviors and non-commercial,temporary homosexual behaviors were risk factors for local infection.

Porcine epidemic diarrhea virus (PEDV) is one of the major etiologies responsible for the acute, highly contagious disease in the digestive tract of pigs, especially neonatal piglets. Since PEDV was first identified in Europe in the late 1970s, it has resulted in significant economic losses in many Asian swine-raising countries, including China. Recently, reverse genetics techniques including targeted RNA recombination, bacteria artificial chromosome system and in vitro ligation have been successfully used to manipulate the genome of PEDV, which providing new strategies for the clear delineation of the functions of the viral proteins, the mechanisms behind PEDV pathogenesis and the design of novel vaccines against PEDV. Here, we review the progresses of different reverse genetics platforms developed for PEDV and their applications, covering the roles of trypsin in PEDV propagation, functions of S and ORF3 protein and the development of next generation PED vaccines, and the perspectives of reverse genetics for PEDV.

Objective To investigate the effect of all-trans retinoic acid (ATRA) on transforming growth factor β1 (TGF-β1)/Notch signaling pathway in injured podocytes induced by adriamycin (ADR) in vitro.Methods Podocytes cultured in vitro were randomly divided into normal group,model group,ATRA treatment control group,12-hour ATRA intervention group and 24-hour ATRA intervention group.Morphological changes were observed by using light microscope.The expressions of TGF-β1,podocin,Notch 1,Jagged 1 mRNA were evaluated through real-time polymerase chain reaction (RT-PCR) and the corresponding proteins were detected by using Western blot.Results (1) No obvious changes between normal group and ATRA treatment control group were revealed as the plump podocytes and distinct outline were found in light microscope,while podocytes in model group showed disordered arrangement,fuzzy boundary,atrophy,hypertrophy and increased cellular debris.Of note,the podocytes in 12-hour ATRA intervention group and 24-hour ATRA intervention group almost returned to normal.(2) In contrast with those in model group,the amounts of TGF-β1,Notchl,Jaggedl mRNA levels decreased in 12-hour ATRA intervention group (1.34 ±0.43 vs.4.16 ±0.31,1.67 ±0.2 vs.4.21 ±0.92,2.08 ±0.27 vs.5.14 ±0.63,q =23.83,11.45,19.67,all P ＜0.05) and 24-hour ATRA intervention group (1.22 ± 0.16 vs.4.16 ± 0.31,1.73 ± 0.53 vs.4.21 ± 0.92,2.08 ± 0.29 vs.5.14 ± 0.63,q =24.85,11.18,19.67,all P ＜ 0.05),and the differences were significant;similar trend was detected in the protein levels (1.04 ± 0.03 vs.4.31 ± 0.10,1.06 ± 0.04 vs.4.47 ± 0.24,1.07 ± 0.04 vs.4.20 ± 0.16,1.06 ±0.03 vs.4.31 ±0.10,1.07 ±0.03 vs.4.47 ±0.24,1.09 ±0.03 vs.4.20 ±0.16,q =163.50,69.61,90.36,162.50,69.40,89.78,all P ＜ 0.05),and the differences were significant;whereas the level of podocin mRNA (1.13 ±0.05 vs.0.40 ± 0.06,1.16 ± 0.03 vs.0.40 ± 0.06,q =36.50,38.00,all P ＜ 0.05) and protein (1.01 ± 0.01 vs.0.44 ±0.01,1.02 ±0.01 vs.0.44 ±0.01,q =180.25,183.41,all P ＜0.05) increased,and the differences were sig nificant.(3) The expressions of Notch1,Jagged1 mRNA were positively correlated with TGF-β1 mRNA (r =0.84,1.00,all P ＜ 0.05),but negatively correlated with podocin mRNA (r =-0.95,-0.94,all P ＜ 0.05) in model group.Conclusions ATRA might alleviate podocyte injury through cutting the expressions of TGF-β1,Notch1,Jagged1 and raising the expression of podocin in injured podocytes induced by ADR.

Objective To investigate the effect of double filtration plasmapheresis (DFPP) upon the removal of donor specific antibody (DSA) in highly sensitized recipients with renal transplantation. Methods Four highly sensitized recipients undergoing renal transplantation received 7 cycles of DFPP. Luminex technology was adopted to monitor the changes of DSA. Clinical efficacy, incidence of acute rejection and adverse reactions were observed. Results After DFPP, the DSA MFI [1036 (0-4113)] was significantly declined than that before treatment [6446 (2999-12905), Z= -2.503, P=0.012]. No hyperacute rejections occurred in four highly sensitized recipients undergoing renal transplantation.Acute rejection was noted in one case, which was mitigated by postoperative DFPP and adjustment of immunosuppressive agents. During postoperative follow-up, the function of transplant kidney was normal and no rejection reactions occurred. The level of albumin was decreased after DFPP. Conclusions DFPP can effectively remove the DSA in the recipients.It is an efficacious and safe approach to prevent the incidence of acute rejections in highly sensitized recipients after renal transplantation.

Retinoic acid (RA) receptor, is a group of ligand-regulated nuclear transcription factors and widely distributed in various tissue cells. The different subtypes of RA receptor can polymerize into homologous or heterogenous dimmers, and then participate in the physiological and pathological processes of embryonic development, cell proliferation and differentiation, and apoptosis. However, the relationship between the expression of RA receptor and the diseases is different in diverse cells or tissues. Recent researches demonstrated that RA receptor is closely related to the pathogenesis and development of renal diseases. This article aims to review the role of the RA receptor and its signal pathway in the development of renal diseases.

Aim To explore the effect of miRNA-143 ( miR-1 4 3 ) on homocysteine ( Hcy ) induced-vascular smooth muscle cells ( VSMCs ) proliferation and the mechanism .Methods VSMCs were cultured and in-cubated with Hcy by using primary cultured method . Then, cells were treated with different concentrations of Hcy and folate .VSMCs proliferation was determined with MTT assay , miR-143 was measured by qRT-PCR, and methylation of miR-143 was determined with meth-ylated PCR.Results After cells were treated with dif-ferent concentrations of Hcy , the proliferation of VSMCs was significantly increased , mRNA expression of miR-143 was decreased and methylation of miR-143 was increased .The proliferation of VSMCs was signifi-cantly decreased when transfected VSMCs with miR-143 precursor , and cell proliferation was increased by using miR-143 inhibitor transfection .Conclusion Hy-pomethylation of miR-143 may inhibit VSMCs prolifera-tion.

Objective To establish a method of gas chromatography with head-space sampling for determination of five residual solvents ( acetone, ethyl acetate, isopropanol, dichloromethane, acetonitrile ) in raw material drug of cefotiam hexetil hydrochloride. Methods Agilent DB-624 capillary column(30 m×0.53 mm,3.0 μm)was used,with FID served as detector and DMF as the solvent. Results Linear relationships were obtained for the 5 residual solvents in their respective concentration ranges ( r=0.999 6-0.999 9,n=5) ,and the detection range was from 0.071 to 0.847μg. The stabilities measured as relative standard deviations ( RSD) for the 5 residual solvents were from 0.40% to 2.12% ( n=3) . The average recovery rates were 99.03% to 103.33%,and RSD were 0.54% to 3.41% ( n=3) . Conclusion The method is simple,sensitive and accurate for the residual solvent analysis in raw material drug of cefotiam hexetil hydrochloride.