Objective:Currently,traditional cervical cancer screening methods,such as high-risk human papillomavirus testing and liquid based cytology(LBC),still possess limitations.This study aims to identify new diagnostic biomarkers to achieve the goal of"precision screening"via exploring the clinical value of DNA methylation[ΔCtP:paired box gene 1(PAX1)and ΔCtJ:junctional adhesion molecule 3(JAM3)]detection in cervical exfoliated cells for the diagnosis of high-grade cervical lesions. Methods:A total of 136 patients who underwent gynecological examinations in the vaginal room of the Department of Gynecology at the Third Xiangya Hospital of Central South University from June 2021 to June 2022 were retrospectively studied.Among them,122 patients had non-high-grade cervical lesions,and 14 patients had high-grade cervical lesions.The variables included general information(age,body mass index,and menopause status),LBC,high-risk human papillomavirus,cervical tissue pathology,vaginal examination results,and the ΔCt values of JAM3 and PAX1 gene methylation.Logistic regression analysis was used to identify the factors affecting the diagnosis of high-grade cervical lesions,followed by correlation analysis and construction of a conditional inference tree model. Results:Logistic regression analysis showed that the methylation ΔCt values of PAX1 and JAM3 genes and LBC detection results were statistically significant between the high-grade cervical lesions group and the non-high-grade cervical lesions group(all P<0.05).Correlation analysis revealed a negative correlation between cervical pathological changes and ΔCtP(r=-0.36,P<0.001),ΔCtJ(r=-0.448,P<0.001),LBC(r=-0.305,P<0.001),or bacterial diversity(r=-0.183,P=0.037).The conditional inference tree showed that when ΔCtJ>10.13,all of patients had non-high-grade cervical lesions,while ΔCtP>6.22,the number of non-high-grade lesions accounted for 97.5%(117/120),and high-grade lesions accounted for only 2.5%(3/120).When ΔCtJ>8.61 and LBC were atypical squamous cell of undetermined significance or negative for intraepithelial lesions or malignancy(NILM),105(99.1%)patients were non-high-grade cervical lesions,only 1(0.9%)patient was high-grade lesion.When the results of LBC were high-grade lesions,only 9 patients'histopathological examination was the high-grade lesions and 3 non-high-grade lesions.When LBC indicated low-grade lesions,atypical squamous cell of undetermined significance,no intraepithelial lesions,and ΔCtP>6.22,117(97.5%)of patients'histopathological examination was the non-high-grade lesions. Conclusion:The JAM3/PAX1 gene methylation test can be used independently for the stratified diagnosis of high-grade/non-high-grade cervical lesions in women with high-risk human papillomavirus infection,independent of the cytological results of cervical excision.The JAM3/PAX1 gene methylation test can also be used in combination with LBC to make up for the shortcomings of low sensitivity of LBC.In addition,the application of methylation kit in large-scale cervical cancer screening in the future will be good to the detection of more patients with high-grade cervical lesions,and achieve early screening and early treatment for cervical lesions/cancer.

Objective To investigate the effect of overexpression of ubiquitin-conjugating enzyme 2T(UBE2T)on radiosensitivity of hepatocellular carcinoma(HCC).Methods Hepa1-6 cells were transfected with a UBE2T-overexpressing or a control lentiviral vector,and the changes in their radiotherapy sensitivity and concentrations of glucose and lactate in the supernatant were assessed using colony-forming assay and colorimetric assay.The transfected cells were inoculated subcutaneously in nude mice or C57BL/6 mice,and tumor growth following irradiation were recorded.The xenografts were collected for analyzing infiltration of CD4+T cells and regulatory T cells(Tregs)using flow cytometry and detecting expressions of HK1 and LDHA using Western blotting.The correlations of UBE2T expression with immune cell infiltration,glycolysis and Tregs in HCC were analyzed using CIBERSORT algorithm and TCGA database,and the results were verified in a co-culture system of Hepa1-6 cells and Tregs.Results UBE2T overexpression caused radiotherapy resistance in both cultured Hepa1-6 cells and xenografts in the tumor-bearing mouse models(especially in C57BL/6 mice).CIBERSORT analysis suggested that a high expression of UBE2T was associated with increased percentages of dendritic cells,T follicular helper cells,M2 macrophages,monocytes,lymphocytes and Tregs in HCC.The UBE2T-overexpressing xenografts showed an increased percentage of Tregs and enhanced expressions of HK1 and LDHA,and irradiation increased infiltration of CD4+T cells and Tregs in the tumor microenvironment.Hepa1-6 cells overexpressing UBE2T showed a decreased glucose concentration and an increased lactate concentration.GSEA analysis suggested that a high UBE2T expression was positively correlated with increased glycolysis and Tregs infiltration in HCC.In the cell co-culture system,UBE2T overexpression significantly enhanced lactate production,proliferation and immunosuppressive functions of Tregs.Conclusion A high UBE2T expression results in radiotherapy resistance of HCC possibly by enhancing glycolysis and cause enrichment of Tregs in the tumor microenvironment.

Objective To investigate the effect of overexpression of ubiquitin-conjugating enzyme 2T(UBE2T)on radiosensitivity of hepatocellular carcinoma(HCC).Methods Hepa1-6 cells were transfected with a UBE2T-overexpressing or a control lentiviral vector,and the changes in their radiotherapy sensitivity and concentrations of glucose and lactate in the supernatant were assessed using colony-forming assay and colorimetric assay.The transfected cells were inoculated subcutaneously in nude mice or C57BL/6 mice,and tumor growth following irradiation were recorded.The xenografts were collected for analyzing infiltration of CD4+T cells and regulatory T cells(Tregs)using flow cytometry and detecting expressions of HK1 and LDHA using Western blotting.The correlations of UBE2T expression with immune cell infiltration,glycolysis and Tregs in HCC were analyzed using CIBERSORT algorithm and TCGA database,and the results were verified in a co-culture system of Hepa1-6 cells and Tregs.Results UBE2T overexpression caused radiotherapy resistance in both cultured Hepa1-6 cells and xenografts in the tumor-bearing mouse models(especially in C57BL/6 mice).CIBERSORT analysis suggested that a high expression of UBE2T was associated with increased percentages of dendritic cells,T follicular helper cells,M2 macrophages,monocytes,lymphocytes and Tregs in HCC.The UBE2T-overexpressing xenografts showed an increased percentage of Tregs and enhanced expressions of HK1 and LDHA,and irradiation increased infiltration of CD4+T cells and Tregs in the tumor microenvironment.Hepa1-6 cells overexpressing UBE2T showed a decreased glucose concentration and an increased lactate concentration.GSEA analysis suggested that a high UBE2T expression was positively correlated with increased glycolysis and Tregs infiltration in HCC.In the cell co-culture system,UBE2T overexpression significantly enhanced lactate production,proliferation and immunosuppressive functions of Tregs.Conclusion A high UBE2T expression results in radiotherapy resistance of HCC possibly by enhancing glycolysis and cause enrichment of Tregs in the tumor microenvironment.

BACKGROUND: SMARCA2 (SWI/SNF Related, Matrix Associated, Actin Dependent Regulator of Chromatin, Subfamily A, Member 2) is an important ATPase catalytic subunit in the switch-sucrose nonfermenting (SWI/SNF) complex. However, its relationship with the pathological features of NSCLC and its prognosis remain unclear. METHODS: We retrospectively reviewed 2390 patients with surgically resected NSCLC, constructed tissue microarrays (TMAs) and performed immunohistochemical assays. We analyzed the correlation of SAMRCA2 with clinicopathological features and evaluated its prognostic value. RESULTS: Among 2390 NSCLC cases, the negative expression ratios of SAMRCA2, SMARCA4, ARID1A, ARID1B and INI1 were 9.3%, 1.8%, 1.2%, 0.4% and 0%, respectively. In NSCLC, male sex, T3 and T4 stage, moderate and poor differentiation, tumor ≥ 2 cm, Ki67 ≥ 15%, SOX-2 negative expression, middle lobe lesion and adenocarcinoma were relative risk factors affecting SMARCA2-negative expression. In lung adenocarcinomas, high-grade nuclei, histological morphology of acinar and papillary, solid and micropapillary and TTF-1-negative expression were relative risk factors affecting SMARCA2-negative expression. Kaplan-Meier survival analysis showed that the OS was shorter in the SMARCA2-negative group. Multivariate survival analysis revealed that SMARCA2-negative expression was an independent factor correlated with a poor prognosis in NSCLC. CONCLUSION In conclusion, SMARCA2-negative expression is an independent predictor of a poor outcome of NSCLC and is a potential target for NSCLC treatment.
Adenosine Triphosphatases/metabolism* ; Carcinoma, Non-Small-Cell Lung/genetics* ; Humans ; Male ; Retrospective Studies ; Transcription Factors/genetics*

【Objective】 To analyze the HIV-, HCV- and HBV- NAT yield rate in different areas of Henan province, so as to provide the basis for disease prevention and control as well as the establishment of a unified quality control standard for nucleic acid testing(NAT) in the Henan province. 【Methods】 The number and prevalence of NAT yielding samples with isolated infectious virus, namely HIV, HCV and HBV, in 18 blood stations in Henan province from 2017~2019, as well as the trends were analyzed. The NAT quality of each laboratory and each testing system was analyzed according to the ratio of reactive individual donation(ID) results to reactive minipools(MP). 【Results】 The HBV, HCV and HIV ID-NAT yield numbers in 3 501 251 blood donations were HBV 2 606(74/100 000), HCV 21 (0.63/100 000), and HIV 34(1.00/100 000). The HBV ID-NAT yield rate showed an upward trend in the whole province from 2017 to 2019, while the prevalence of HIV and HCV ID-NAT yield didn′t differ significantly during three years. 5 kinds of NAT detection systems were applied in 18 blood centers. among which Ⅰ, Ⅱ, Ⅳ and Ⅴ were triplex detection systems. 2661 ID-reactive samples were implicated in 5 595 MP-reactive samples, with a resolution rate of 47.56%. The resolution rate of triplex NAT system Ⅰ, Ⅱ, Ⅳ and Ⅳ was 39.63%~47.95%, 40.43%~54.36%, 51.61% and 70.00%~45.45%, respectively. An upward trend in triplex NAT resolution rate was observed in 8 laboratories, i. e.B, D, E, F, I, K, L and Q, and an descending trend in A and C. The NAT system Ⅲ, a ID-NAT system, was used only by laboratory C, presenting a NAT-yield rate of 0.19% (282/145 474) and resolution rate of 46.45% (131/282). 【Conclusion】 The majority of NAT-yield of one infectious virus in Henan province is HBV, presenting annual increasing trend. The quality management of NAT laboratories should be strengthened as the divergence was seen in the performance of different NAT laboratories.

【Objective】 To analyze the cause of single-ELISA reactive of four blood screening items in 18 blood stations in Henan, so as to provide the basis for improving the quality of blood screening. 【Methods】 The single-ELISA reactive rate of HBsAg, anti-HCV, HIV Ag/Ab and anti-TP of 18 blood station laboratories in Henan throughout 2019 was calculated, and the causes were analyzed according to different ELISA reagent combinations and gray area settings in each laboratory. 【Results】 The overall single-ELISA reactive rates of HBsAg, anti-HCV, HIV Ag/Ab and anti-TP were 1.740(2 154/1 237 789), 0.564‰(698/1 237 789), 1.421‰(1 759/1 237 789) and 1.561‰(1 932/1 237 789), respectively, showing significant differences by detection items (P <0.05). Person correlation analysis showed that the single-ELISA reactive rate was independent of the gray area settings.but dependent on laboratories and reagent combinations. The single-ELISA reactive rate of HBsAg, anti-HCV, HIV Ag/Ab and anti-TP in D laboratory was the highest and higher than that in other labs using the same reagent.The laboratories with high HBsAg single-ELISA reactive rate were mostly those using a combination of imported reagents and domestic reagents, including the top 6 laboratories. The laboratories with high anti-HCV single-ELISA reactive rate were mostly those using certain domestic reagents. No obvious rules was noticed by single-ELISA reactive for anti-HIV. Laboratories with high anti-TP single-ELISA reactive rate were mostly those using combination 4. 【Conclusion】 The HBsAg single-ELISA reactive rate was the highest in the four blood screening items of blood station laboratories in Henan. The single-ELISA reactive rate is related to the laboratory itself and the reagent manufacturer, suggesting that laboratory quality control should be strengthened and proper reagent combination should be selected to reduce the waste of blood.