Objective To explore the value of peripheral blood circulating DNA in predicting the efficacy and prognosis of immunotherapy for advanced non-small cell lung cancer.Method A retrospective study was conducted on 78 NSCLC patients who were admitted to the Respiratory and Critical Care Medicine Department of the First Affiliated Hospital of Hebei North University and were treated with tirelizumab for advanced driver gene negativity from January 2021 to December 2021.After 2 cycles of immunotherapy,the efficacy was evaluated according to the Solid Tumor Efficacy Evaluation Criteria(RECIST 1.1),including complete remission,partial remission,disease stability,and disease progression.CR and PR patients were defined as the experimental group(n=48)Other patients were defined as the control group(n=30),and the ctDNA levels in peripheral blood were measured before and after treatment in both groups.ROC curves were used to analyze the predictive value of periph-eral blood ctDNA levels for achieving objective remission after immunotherapy.All patients were followed up and their progression free survival were calcutated.Using univariate and multivariate regression analysis identified the factors affecting the prognosis of patients after immunotherapy.Using Spearman correlation coefficient analyzed the correlation between ctDNA levels and PFS.Kalplan Meier survival curve were used for survival analysis.Result The peripheral blood ctDNA levels before and after treatment in the experimental group were(4.47±1.21)ng/μL and(2.65±1.14)ng/μL,respectively(t=7.559,P<0.001),while those in the control group were(4.54±1.15)ng/mL and(4.29±1.57)ng/μL,respectively(t=0.699,P=0.487).There was no statistically significant difference in peripheral blood ctDNA levels between the two groups before treatment(t=-0.25,P=0.801).The peripheral blood ctDNA levels in the experimental group decreased compared to the control group after treatment(t=-5.35,P<0.001).The ROC curve analysis showed that the area under the curve for predicting objective remission after immunotherapy based on peripheral blood ctDNA levels was 0.819,with a sensitivity of 81.3%and specificity of 80%.Peripheral blood ctDNA levels were negatively correlated with progression free survival(r=-0.784,P=0.000).Single factor COX regression was used to analyze the clinical and pathological characteristics and ctDNA levels of enrolled patients,and the results showed that the maximum tumor diameter was greater than 5 cm(HR=0.501,95%CI:6.731～35.567)Tumor stage IV(HR=0.392,95%CI:0.227～0.677),treatment approach(HR=15.473,95%CI:6.731～35.567),and ctDNA levels(HR=4.657,95%CI:3.182～6.555)are all influencing factors for PFS in advanced NSCLC patients after immunotherapy.Multiple factor analysis was conducted on the appeal indicators with statistical differences,and the results showed that treatment approach(HR=2.981,95%CI:1.019～8.722)and peripheral blood ctDNA levels(HR=3.918,95%CI:2.619～5.861)It is an independent influencing factor of PFS in advanced NSCLC patients.The Kalplan Meier survival curve was used for analysis,and the results showed that the median PFS of the treatment effective group was 8.4 months,while the median PFS of the control group was 5.4 months.(χ2=49.277,P=0.000).Conclusion Immunotherapy combined with chemotherapy can enhance the ability to kill tumor cells,and peripheral blood ctDNA levels can evaluate the efficacy and prognosis of immunotherapy,which can be used to guide immunotherapy in advanced NSCLC patients.

Objective To investigate the correlation between the expression of long non-coding ribonucleic acid growth arrest specific 5(lncRNA GAS5),phospholysine phosphohistidine inorganic pyrophosphate phos-phatase(LHPP)and epithelial-mesenchymal transition(EMT)in cancer tissues of patients with non-small cell lung cancer(NSCLC)and its clinical significance.Methods Cancer tissues and adjacent tissues of 90 patients with NSCLC who underwent surgical resection in the First Hospital Affiliated to Hebei North College from June 2018 to January 2020 were collected.The expressions of lncRNA GAS5,LHPP and EMT-associated pro-teins[E-calmodulin(E-Cad),N-calmodulin(N-Cad),and vimentin(VIM)]were detected by real-time fluores-cence quantitative polymerase chain reaction.The relationship between lncRNA GAS5 and LHPP mRNA and clinicopathological features in cancer tissues of NSCLC patients was analyzed,and the correlation between ln-cRNA GAS5 and LHPP mRNA and EMT-associated proteins expression in cancer tissues of NSCLC patients was analyzed by Pearson correlation.Kaplan-Meier method was used to plot the survival curves of NSCLC pa-tients with different lncRNA GAS5 and LHPP mRNA expressions,and multivariate Cox regression was used to analyze the prognostic factors of NSCLC patients.Results The expressions of lncRNA GAS5,LHPP mR-NA and E-Cad mRNA in cancer tissues of NSCLC patients were lower than those in adjacent tissues,while the expressions of N-Cad mRNA and VIM mRNA were higher than those in adjacent tissues,with statistical sig-nificance(P＜0.05).Pearson correlation analysis showed that lncRNA GAS5 in cancer tissues of NSCLC pa-tients was positively correlated with E-Cad mRNA expression(r=0.724,P＜0.001),and negatively correla-ted with N-Cad mRNA and VIM mRNA expression(r=-0.699,-0.689).P＜0.001);lncRNA GAS5 was positively correlated with LHPP mRNA expression(r=0.651,P＜0.001).The mRNA expressions of ln-cRNA GAS5 and LHPP in cancer tissues of NSCLC patients with different degrees of differentiation,tumor TNM stage and lymph node metastasis were significantly different(P＜0.05).Kaplan-Meier survival curve a-nalysis showed that the 3-year overall survival rate in the lncRNA GAS5 high expression group[68.18％(30/44)]was higher than that in the lncRNA GAS5 low expression group[36.96％(17/46)].The 3-year overall survival rate in the high LHPP mRNA expression group[67.39％(31/46)]was higher than that in the lowLHPP mRNA expression group[36.36％(16/44)],and the difference was statistically significant(x2=10.274,10.322,P＜0.05).Low differentiation,TNM stage Ⅲ and lymph node metastasis were independent risk factors for death in NSCLC patients,and lncRNA GAS5≥1.32 and LHPP mRNA≥1.12 were independ-ent protective factors(P＜0.05).Conclusion The low expression of lncRNA GAS5 and LHPP mRNA in cancer tissues of patients with NSCLC is related to EMT-associated proteins expression,differentiation de-gree,tumor TNM stage,lymph node metastasis and prognosis,and may become a new target for the diagnosis and treatment of NSCLC.

Objective:To investigate the effect and mechanism of miR-217 targeted regulation of extracellular signal-regulated kinase 2(ERK2)expression on activity and immune escape of non-small cell lung cancer cells(NSCLC).Methods:qRT-PCR was used to detect expression levels of miR-217 and ERK2 mRNA in NSCLC tissues,adjacent tissues,and HLF-1,A549 and HCC827 cell lines.Analyzed prognosis and survival status of NSCLC patients with different miR-217 expression level.Bioinformatics and dual luciferase gene reporting experiments were used to analyze the targeting relationship between miR-217 and ERK2.Cultivated NSCLC A549 cells and divided them into NC group,miR-217 inhibitor group,miR-217 mimic group,miR-217 mimic+ERK2 NC group and miR-217 mimic+ERK2 group.Except for the NC group without any treatment,all other groups were transfected with corresponding plasmids to analyze the proliferation activity and immune escape status of A549 cells in each group,and clarified the mechanism of action.Results:Compared with adjacent tissues,expression of miR-217 in NSCLC tissue was decreased,while expression of ERK2 mRNA was increased(P<0.05).Compared with human normal lung fibroblast HLF-1 cell lines,expression of miR-217 in NSCLC cell lines A549 and HCC827 were decreased,while expression of ERK2 mRNA was increased(P<0.05).Analysis of the relationship be-tween miR-217 and prognosis of NSCLC patients based on Kaplan-Meier Plotter database showed that low expression of miR-217 was associated with poor prognosis of patients(HR=0.90,P=0.033).Dual fluorescein reporter genes showed matching sequences between the 3'UTR regions of miR-217 and ERK2.miR-217 mimic fragment could inhibit ERK2-WT signal,but had no effect on ERK2-MUT.Compared with NC group,cell proliferation activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 inhibitor group were in-creased,while CD8+T cell activity was decreased,and cell proliferation activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 mimic group were decreased,while CD8+T cell activity was increased(P<0.05).Compared with miR-217 mimic group,cell pro-liferation activity,CD8+T cell activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 mimic+ERK2 NC group had no signifi-cant changes(P>0.05),cell proliferation activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 mimic+ERK2 group were increased,while CD8+T cell activity was decreased(P<0.05).Conclusion:Overexpression of miR-217 can reduce the activity of NSCLC cell A549,inhibit the expression of PD-L1,activate CD8+T cells in tumor microenvironment,and then inhibit immune es-cape,which may play a role by targeting ERK2.

Objective:To investigate five kinds of tuberculin skin test (TST), tubercle bacillus-antibody(TB-Ab), interferon-γ release assay(IGRA), tubercle bacillus-DNA (TB-DNA) and collection of bacterial centrifugal smears methods, the application value of combined detection in improving the diagnostic efficiency of pneumoconiosis complicated with tuberculosis.Methods:A total of 150 eligible patients with pneumoconiosis from January 2018 to January 2019 in Zhangjiakou Pulmonary Hospital were selected as the research subjects, and all of them underwent TST, TB-Ab, IGRA, TB-DNA and bacterial centrifugal smear detection. Compared the positive rates of five detection methods in pneumoconiosis and its different stages, and compare the proportion of tuberculosis infection and tuberculosis in different stages of pneumoconiosis.Results:Among the 150 patients with pneumoconiosis, 41 cases (27.33%) were with pneumoconiosis complicated with tuberculosis infection, 24 cases (16.00%) with pneumoconiosis complicated with clinically diagnosed pulmonary tuberculosis, 21 cases (14.00%) with pneumoconiosis complicated with confirmed pulmonary tuberculosis, and 45 cases (30.00%) with pneumoconiosis complicated with pulmonary tuberculosis; with the improvement of pneumoconiosis stage, the proportion of pneumoconiosis combined with tuberculosis infection and pulmonary tuberculosis increased significantly ( P<0.05). Compared with TB-Ab, TB-loop-mediated isothermal amplification(LAMP), and interlayered cup collection centrifuge smear method, the overall positive rate of IGRA detection and pneumoconiosis stage Ⅲ were higher ( P<0.05), but there was no significant difference compared with TST detection ( P>0.05). The positive rate of combined detection was higher, but there was no significant difference compared with IGRA detection ( P>0.05). With the increase of pneumoconiosis stage, the positive reaction intensity of TST decreased, and the positive value of TB-Ab and IGRA increased. Conclusions:The combined detection of TST, TB-Ab, IGRA, TB-DNA and bacterial centrifugal smear method can significantly improve the diagnostic efficiency of pneumoconiosis combined with tuberculosis.

OBJECTIVE：To investigate the efficacy of hormone combined with cyclophosphamide in the treatment of connective tissue disease-associated interstitial lung disease （CTD-ILD）and to analyze its influential factors. METHODS ：100 patients diagnosed as CTD-ILD in our hospital from Jan. 2018 to Jan. 2019 were randomly divided into observation group and control group ，with 50 cases in each group. Control group was treated with Compound cyclophosphamide tablets ，50 mg each time，3-4 times each day. Observation group was additionally treated with Prednisone acetate tablets ，10 mg each time ，3-4 times each day ，on the basis of control group. Treatment courses of 2 groups lasted for 6 months. The clinical efficacy ，the occurrence of ADR，lung function before and after treatment ，the levels of peripheral IL- 6，CRP and PCT and quality of life （SGRQ score ）were compared between 2 groups. According to the therapeutic efficacy ，all patients were divided into effective group and ineffective group. The related factors influencing the clinical efficacy of CTD-ILD were analyzed by univariate and multivariate Logistic regression analysis. RESULTS ：After treatment ，total response rate ，FVC，FEV1 and DLCO of observation group were significantly higher than those of control group ，while SGRQ score ，levels of IL- 6，CRP and PCT in peripheral blood were significantly lower than control group （P＜0.05）. There was no significant difference in the total incidence of ADR between 2 groups（P＞0.05）. Univariate analysis showed that there were no significant differences in gender ，age，past medical history and CTD type between effective group and ineffective group （P＞0.05）. However ，there were statistical significancant differences in the distribution of different levels of IL- 6，CRP and PCT in peripheral blood between 2 groups（P＜0.05）. Multivariate Logistic regression analysis showed that IL- 6 was an independent risk factor for therapeutic efficacy of combined therapy [OR （95％CI）＝ 4.537（3.668，10.352），P＝0.002]. CONCLUSIONS ：Hormone combined with cyclophosphamide can significantly improve the therapeutic efficacy of CTD-ILD patients ，improve their lung function and quality of life ，and reduce the expression level of inflammatory factors. The level of IL- 6 is an independent risk factor affecting the efficacy of the treatment ，and its changes should be paid close attention to during the treatment.

目的：探究环状 RNA(circular RNA, circRNA）0072088 在非小细胞肺癌（non-small cell lung cancer，NSCLC）细胞中 的生物学功能及其作用机制。方法：在公共基因芯片数据库 Gene Expression Omnibus（GEO）中下载 GSE101684 数据集，通过 GEO2R 分析得到差异基因。通过 qPCR 检测 NSCLC 细胞 H165、H358、H460、H226 和 A549 细胞中 circ_0072088 的表达水平， 随后采用 CCK-8 法和 Transwell 小室法分别检测 circ_0072088 对 NSCLC 细胞增殖、迁移和侵袭的作用。通过 CircInteractome 和 TargetScan 数据库预测 circ_0072088 与 miR-545-3p、miR-545-3p 与 STAT3 之间的靶向关系，并通过双荧光素酶报告基因实 验和 RNA 结合蛋白免疫沉淀（RNA binding protein immunoprecipitation，RIP）实验验证 circ_0072088、miR-545-3p 与 STAT3 之 间的靶向关系。结果：circ_0072088 在 NSCLC 细胞系中的表达均显著上调（均 P<0.05）。过表达 circ_0072088 促进了 NSCLC 细胞的增殖、侵袭和迁移（均 P<0.05）；敲低 circ_0072088 抑制了 NSCLC 细胞的增殖、侵袭和迁移（均 P<0.05）。miR-545-3p 是 circ_0072088 的下游靶点，可以被 circ_0072088 吸附；STAT3 是 miR-545-3p 的靶基因，可以被 circ_0072088 间接正向调控。 结论：Circ_0072088 通过调节 miR-545-3p /STAT3 轴促进 NSCLC 细胞的增殖和转移。

BACKGROUND:High expression of stem cell transcription factors, Sox2 and Oct4, has been confirmed to be closely related to the occurrence of lung cancer.OBJECTIVE:To explore the expression of Sox2 and Oct4 in lung cancer tissues and its correlation with microvascular generation and clinical-pathologic features.METHODS:The expression levels of Sox2 and Oct4 and microvessel density (MVD) in lung cancer tissues (60 cases) and normal tissues (60 cases) were examined by immunohislochemistry. Then, the correlation of the expression of Sox2 and Oct4 and MVD value with clinical-pathologic features in lung cancer was analyzed.RESULTS AND CONCLUSION:The positive expression of Sox2 and Oct4 was 46.67% (28/60) and 71.67% (43/60) in lung cancer tissues, respectively, while Sox2 and Oct4 were negatively expressed in normal tissues, and there was a significant difference between lung cancer and normal tissues (P < 0.001). The MVD value was 16.22±2.18 in lung cancer tissues, which was significantly higher than that in normal tissues (4.36±2.07) (P < 0.001). Expression of Sox2,Oct4 and MVD value were correlated with TNM stage, differentiation degree, ymph node metastasis, vessel invasion,and liver metastasis (all P < 0.05), but not with the patient's age and gender (both P > 0.05). The MVD value with Sox2 and Oct4 positive expression was significantly higher than that with Sox2 and Oct4 negative expression (P < 0.001).Spearmen analysis results showed that there was no correlation between Sox2 and Oct4 in lung cancer tissues (r=2.752,P > 0.05). To conclude, the high expression of Sox2 and Oct4 has a high correlation with the initiation and progression of lung cancer, and has positive correlation with the MVD, both of which are significantly concerned with the microvascular generation, invasion and hematogenous metastasis of lung cancer.

Adult ; Air Pollutants, Occupational ; analysis ; Dust ; analysis ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Occupational Health ; Welding ; Young Adult

To evaluate PCR-sequencing for clinical detection of human papillomavirus (HPV) genotypes in cervical cell specimens, we applied PCR-sequencing to HPV detection and genotyping by general primer PGMY09/11, which targets the HPV most conserved L1 gene. Samples with multiple infections were subjected to HPV type-specific PCR. Among the 325 cervical samples, 228 were HPV positive, of which 66 showed multiple infections. In all, 27 different HPV genotypes were identified, with HPV 16 being the most prevalent, followed by HPV 58 and 52. The prevalence of high-risk HPV infection increased with the severity of cervical lesions (P < 0.05), whereas the proportion of multiple infections declined significantly from LSIL to SCC (P < 0.05). Both rates of overall and high-risk HPV infection were the highest in 21-30 age groups. There was substantial agreement between the HC2 and PCR-sequencing assay for detection of high-risk HPV (kappa = 0.675). PCR-sequencing was effective in HPV detection and genotyping, and it could be potentially applied to large scale HPV screening.
Adult ; Aged ; Cervix Uteri ; pathology ; virology ; China ; epidemiology ; Female ; Genotype ; Human papillomavirus 16 ; isolation & purification ; Humans ; Middle Aged ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; epidemiology ; virology ; Polymerase Chain Reaction ; methods ; Vaginal Smears ; Young Adult

We selected and characterized isolates of Lactobacillus crispatus (L. crispatus) for potential preventing infections of the female reproductive tract. We cultured vaginal swabs from healthy volunteers on de Man, Rogosa and Sharpe (MRS) agar and identified the isolates at the species level by 16S rRNA sequence and genotyped the isolates of Lactobacillus by PCR amplification of repetitive bacterial DNA elements (rep-PCR). Furthermore, 10 L. crispatus strains were assessed for hydrogen peroxide (H2O2) and acid production. Overall 65 isolates were confirmed to be Lactobacillus by sequence analogy, among them 19 were L. crispatus, 17 were Lactobacillus jensenii and 12 were Lactobacillus fermentum. rep-PCR produced specie and strain-specific genomic fingerprints for the Lactobacillus isolates. The selected 10 L. crispatus isolates produced highly acidic environment after growth in MRS. The isolates T22-3 and T29-5 demonstrated high production of H2O2. This study indicated that there are individual differences with vaginal Lactobacillus colonization, and strain diversity within vaginal L. crispatus isolates, T22-3 and T29-5 might be candidates for restoring urogenital health environment in females.
Adult ; Female ; Genotype ; Humans ; Hydrogen Peroxide ; metabolism ; Interspersed Repetitive Sequences ; Lactobacillus ; classification ; genetics ; isolation & purification ; physiology ; Polymerase Chain Reaction ; methods ; Vagina ; microbiology