Objective:To investigate the effect and mechanism of miRNA-5193 (miR-5193) on the sensitivity of cervical cancer Caski cells to cisplatin.Methods:The expression of miR-5193 in cervical cancer cell lines C33A, SiHa, Caski and normal cervical cell line Ect1/E6E7 were determined by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Caski cells were divided into control group (no transfection, normally cultured), miR-5193-negative control (miR-NC) group (transfected with miR-NC mimic), miR-5193 group (transfected with miR-5193 mimic), miR-NC+cisplatin group (transfected with miR-NC mimic and treated with 10 μg/ml cisplatin), miR-5193+cisplatin group (transfected with miR-5193 mimic and treated with 10 μg/ml cisplatin), miR-5193+cisplatin+NC group (cotransfected with Foxp3-negative control vector and miR-5193 mimic, and treated with 10 μg/ml cisplatin), and miR-5193+cisplatin+Foxp3 group (cotransfected with Foxp3 overexpression vector and miR-5193 mimic, and treated with 10 μg/ml cisplatin). Proliferation was detected by methyl thiazolyl tetrazolium (MTT), cell cycle was detected by PI single staining method, cell apoptosis was detected by Annexin V-FITC/PI double staining method, and expressions of CDK2, p27 and C-caspase-3 proteins in cells were detected by Western blot. Bioinformatics software was used to predict miR-5193 target genes, and the luciferase reporting system was used to identify the targeting relationship.Results:The relative expression of miR-5193 in cervical cancer C33A, SiHa and Caski cells was lower than that in normal cervical Ect1/E6E7 cells (0.56±0.06, 0.41±0.03, 0.23±0.02 vs. 1.00±0.10, all P < 0.05). Compared with the control group and miR-NC group, the cell proliferation activity (absorbance value) in miR-5193, miR-NC+cisplatin and miR-5193+cisplatin groups decreased (0.58±0.06, 0.59±0.07 vs. 0.38±0.04, 0.40±0.05, 0.23±0.02, all P < 0.05), the cell apoptosis rate increased [(2.5±0.2)%, (2.7±0.3)% vs. (12.6±1.2)%, (11.9±1.5)% , (18.9±1.7)%, all P < 0.05], and the proportion of cells in G 0/G 1 phase increased [(50.4±4.2)%, (51.3±6.3)% vs. (62.3±3.2)%, (61.9±5.8)%, (71.4±5.4)%, all P < 0.05]. The expression levels of p27 and C-caspase-3 proteins increased, and the expression level of CDK2 protein decreased. The software predicted that the target gene of miR-5193 was Foxp3, which was confirmed by the luciferase reporting system. Compared with the miR-5193+cisplatin+NC group, the cell proliferation activity (absorbance value) in miR-5193+ cisplatin+Foxp3 group increased (0.24±0.03 vs. 0.65±0.05, t = 21.094, P < 0.01), the proportion of cells in G 0/G 1 phase decreased [(71.0±6.4)% vs. (60.3±4.1)%, t = 4.196, P < 0.01], the apoptosis rate of cells decreased [(19.6±1.6)% vs. (11.5±1.2)%, t = 11.880, P < 0.01], the expression levels of p27 and C-caspase-3 proteins in cells decreased, and the expression levels of CDK2 and Foxp3 proteins increased. Conclusion:The miR-5193 may increase the sensitivity of cervical cancer Caski cells to cisplatin in vitro by targeted inhibition of the Foxp3 gene.

Objective To study the endogenous metabolites of liver-kidney deficiency syndrome in knee-joint osteoarthritis (KOA) and explore the metabolic profile of KOA liver-kidney deficiency syndrome. Methods Totally 50 cases of KOA with liver-kidney deficiency syndrome and 50 cases of KOA with non-liver-kidney deficiency syndrome were collected respectively, and 20 cases of healthy volunteers were collected as the normal group. The serum samples of subjects were collected after fasting for 8 h. Hydro-Nuclear Magnetic Resonance (1H-NMR) spectrometer was collected. Principal component analysis and partial least squares discrimination analysis were used to conduct multivariate statistical analysis. Results 1H-NMR could identify 23 kinds of metabolites, and there was statistical significance between KOA patients and healthy volunteers (P<0.05, P<0.01). There was statistical significance in cartilage matrix metabolism, energy metabolism, lipid metabolism, pain, inflammation-related metabolites in patients with KOA liver-kidney deficiency syndrome and patients with KOA non-liver-kidney deficiency syndrome (P<0.05). Conclusion Patients with KOA liver-kidney deficiency syndrome have a unique 1H-NMR metabolic profile, KOA syndrome has a metabolic material basis.

Hexanes ; adverse effects ; Humans ; Occupational Injuries ; chemically induced ; Pyrroles ; adverse effects ; Trauma, Nervous System ; chemically induced

Objective To explore the association of COX-2 Gly587Arg polymorphism with the risk of primary liver cancer.Methods Two hundred and seventy patients with primary liver cancer and 540 health people were selected as our subjects.DNA were extracted from peripheral blood lymphocytes,and genotypes were measured by polymerase chain reaction-restriction fragment length polymorphism method.Odds ratios(OR) and 95％ confidence intervals(CI) were estimated by logistic regression.Results Two kinds of genotype (587Gly/ Gly and Gly/Arg) were found in all participants.No one carried 587Arg/Arg genotype.Among primary liver cancer patients,91.5％ (247/270,) 8.5％ (23/270) of individuals carried 587Gly/Arg and Gly/Arg genotype,which was significantly higher than that of controls (96.5％ (521/540,) 3.5％ (19/540)).Multivariate Logistic regression analysis showed that individual carried 587Gly/Arg genotype had an increased risk of developing primary liver cancer (OR =2.56,95％ CI =1.37-4.79,P =0.003) compared with 587Gly/Gly carriers.Conclusion COX-2 Gly587Arg polymorphism is a risk factor for primary liver cancer in Han.

Objective:To study the amino acid and codon usage profile of HIV-1 Env gene in donors whose serum exhibit highly broad cross-neutralizing activity. Methods:The samples were divided into highly broad cross-neutralizing activity group (hBCN + group) and non-highly broad cross-neutralizing activity group (hBCN - group) based on whether the neutralization breadth was higher than 90% or not. Full-length Env genes were amplified by single genome amplification (SGA) method from patients′ plasma samples, and the characteristics of Env sequences in hBCN + group were compared with hBCN - group. The correspondence analysis (COA) on relative amino acid usage (RAAU), adaptability to host based on similarity index D( A, B) and relative synonymous codon usage (RSCU) values of Env genes (hBCN + and hBCN -) with respect to human host RSCU were analyzed. Results:Correspondence analysis showed that the RAAU data of hBCN + group and hBCN - group were distributed along the two main axes to form two relatively separated clusters, indicating that the Env genes of the two groups had relatively unique amino acid usage patterns; the similarity index calculation results showed that hBCN + group (0.097) was lower than the hBCN - group (0.102), in addition, the Env gene of the hBCN + group had less frequency of similarly selected codons with human host system compared to hBCN - group. Conclusions:Env genes in hBCN + group and hBCN - group may have relatively unique amino acid usage patterns, and virus strains in hBCN + group are less adaptable to the host than those in hBCN - group.

Objective:To investigate the relationship between CD8 +FoxP3 +CD25 + T cell subsets and the therapeutic effect of programmed death receptor 1 (PD-1) inhibitor pembrolizumab in treatment of uterine cervical cancer. Methods:The data of 105 patients with uterine cervical cancer who received pemblizumab therapy based on chemotherapy in the First Hospital of Qinhuangdao from January 2018 to January 2020 were retrospectively analyzed. Flow cytometry was used to detect the ratio of CD8 +FoxP3 +CD25 + T cell in peripheral blood of patients. The efficacy and safety were analyzed. According to the efficacy, all patients were divided into remission group (complete remission + partial remission) and non-remission group (stable disease + progressive disease). The clinical characteristics and CD8 +FoxP3 +CD25 + T cell ratio of the two groups were compared. Multivariate logistic regression model was used to analyze the influencing factors for the efficacy. The efficacy of CD8 +FoxP3 +CD25 + T cell ratio predicting the therapeutic effect of patients was analyzed by using receiver operating characteristic (ROC) curve. Results:The objective remission rate of all patients was 17.14% (18/105), and the incidence of adverse reaction was 39.05% (41/105). The proportion of patients with a family history of cervical cancer in the remission group was lower than that than in the non-remission group [5.56% (1/18) vs. 34.48% (30/87)], and the difference was statistically significant ( χ2=6.00, P=0.014). The proportion of CD8 +FoxP3 +CD25 + T cell of 105 patients before and after treatment was (0.83±0.21)% and (0.77±0.10)%, respectively; the proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the remission group was (0.55±0.26)%, (0.31±0.12)%, respectively; the proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the non-remission group was (0.89±0.30)%, (0.87±0.28)%, respectively. The proportion of CD8 +FoxP3 +CD25 + T cell after treatment in the remission group was lower than that before treatment ( P < 0.05); there was no statistically significant difference in the proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the non-remission group ( P>0.05). The proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the non-remission group was higher than that in the remission group (all P<0.001). The proportion of CD8 +FoxP3 +CD25 + T cell higher than the mean value of both groups before treatment and the proportion of CD8 +FoxP3 +CD25 + T cell higher than the mean value of both groups after treatment were independent risk factor of disease remission ( OR=2.542, 95% CI 1.649-3.918, P<0.001; OR=2.936, 95% CI 2.154-4.002, P<0.001). ROC curve analysis showed that the area under the curve of CD8 +FoxP3 +CD25 + T cell ratio predicting the disease remission before treatment was 0.720, and its best cut-off value was 0.77%, the senfitivity was 77.78%, the specificity was 70.11%. Conclusions:Early detection of CD8 +FoxP3 +CD25 + T cell ratio helps to predict the effect of PD-1 inhibitor pembrolizumab therapy for uterine cervical cancer.

Objective:To investigate the role of follicular helper T(Tfh) cells and galactose deficiency IgA 1(Gd-IgA 1) in the children that were suffering from Henoch-Sch?nlein purpura(HSP) and Henoch-Sch?nlein purpura nephritis(HSPN)and the correlation between them. Methods:According to the presence or absence of renal injury, 62 children with HSP were divided into HSP group with 32 children and HSPN group with 30 children.Twenty children who underwent physical examination at outpatients were known as the healthy control group.Flow cytometry was used to measure the proportion of Tfh(CD4 + CXCR5 + PD-1 + ) in peripheral blood.Immunoturbidimetry and ELISA were used to measure the serum levels of IgA 1 and Gd-IgA 1 respectively. Results:(1) The proportion of Tfh cells in peripheral blood and the serum levels of Gd-IgA 1 in both HSP group and HSPN group had significantly increased than those in healthy control group( P<0.01). Compared result of the HSPN group with HSP group, the proportion of Tfh cells in peripheral blood and the serum levels of Gd-IgA 1 in HSPN group were higher than that in HSP group( P<0.05). (2) In the HSPN group, the proportion of peripheral blood Tfh cells and the serum levels of Gd-IgA 1 in group of renal pathology ≥ grade Ⅲ and heavy proteinuria were significantly elevated compared with group of renal pathology < grade Ⅲ and non-heavy proteinuria(<0.01). (3) In the healthy control group, the serum levels of Gd-IgA 1 was positively correlated with the proportion of Tfh cells in peripheral blood and the serum levels of Gd-IgA 1( P<0.05). Conversely, a non-positive correlation was shown in HSP and HSPN groups( P>0.05). Conclusion:The excessive activation of Tfh cells and the serum levels of Gd-IgA 1 may be one of the pathogenesis of HSP/HSPN, the degree of increment of the two factors may be related to the activity and severity of the disease.The mechanism of Tfh cells potentially leading to an increase of Gd-IgA 1 production requires further study.

Objective:To investigate the clinical effect of programmed death receptor-1 (PD-1)/programmed death receptor ligand-1 (PD-L1) immunotherapy combined with concurrent radiotherapy and chemotherapy in the treatment of locally advanced cervical cancer (LACC).Methods:From November 2018 to October 2019, 51 LACC patients in Qinhuangdao First Hospital who received anti-PD-1/PD-L1 immunotherapy (pembrolizumab) combined with concurrent radiotherapy and chemotherapy [intensity modulated radiotherapy (IMRT)+ TP (taxol+ carboplatin) chemotherapy] were selected as the observation group. 51 LACC patients who received concurrent chemotherapy and radiotherapy were selected as the control group. The objective remission rate, disease control rate, tumor markers [squamous cell carcinoma antigen (SCCAg), soluble cytokeratin 19 fragment (CYFRA21-1), and carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125)], proliferation and apoptosis indicators [survivin (Survivin), B-cell lymphoma-2 (Bcl-2), Caspase-3 (Caspase-3), apoptosis-promoting substance (Bax)], PD-1/PD-L1 [soluble PD-L1 (sPD-L1), CD4 + T cell surface PD-1 expression (PD-1 CD4 + T cells), CD8 + T cell surface PD-1 expression (PD-1 CD8 + T cell) and CD14 + monocyte surface PD-L1 expression (PD-L1 CD14 + monocyte)], safety and survival rate within 1 year were compared between the two groups. Results:(1) Disease control and safety: the objective response rate and disease control rate of the observation group were 80.39%(41/51) and 92.16%(47/51), respectively, which were higher than those of the control group by 39.22%(20/51) and 70.59%(36/51) (all P<0.05), but there was no significant difference in the incidence of side effects between the groups (all P>0.05). (2) Tumor markers and proliferation and apoptosis indexes: compared with those before treatment, the levels of serum SCCAg, CYFRA21-1, CEA, CA125, survivin and Bcl-2 in the two groups after treatment were significantly lower, and the levels of Caspase-3 and Bax were significantly higher; the above indexes in the observation group were better than those in the control group after treatment (all P<0.05). (3) PD-1/PD-L1: after treatment, sPD-L1, PD-1 CD4 + T cells, PD-1 CD8 + T cells and PD-L1 CD14 + monocytes in the observation group were significantly lower than those before treatment (all P<0.05). After treatment, the sPD-L1, PD-1 CD4 + T cells, PD-1 CD8 + T cells, PD-L1 CD14 + monocytes in the observation group were lower than those in the control group (all P<0.05). (4) Survival: the survival rate of the observation group was higher than that of the control group within 1 year ( P<0.05). Conclusions:The clinical effect of anti-PD-1/PD-L1 immunotherapy combined with concurrent radiotherapy and chemotherapy in the treatment of LACC is significant. It can effectively inhibit the progression of the disease by regulating tumor markers, proliferation and apoptosis indicators and PD-1/PD-L1 expression without increasing the risk of treatment, and has a positive effect on improving the survival rate of patients.

Objective To investigate the correlation between serum vitamin D level in human body and Chlamydia trachomatis (Ct) genitourinary tract infection. Methods This study enrolled 174 outpa-tients (male: 95, female: 79, 20-49 years) infected with Ct and 380 healthy subjects (male: 211, female:169, 20-49 years) in Tianjin from November 1, 2016 to March 15, 2017. Blood samples were collected from all subjects after fasting overnight and the time points for sample collection in the Ct infection group were before and after a course of antibiotic treatment. Serum 25-hydroxyvitamin D [25-(OH)D] levels were measured with enzyme-linked immunosorbent assay (ELISA). PCR assay was used to assess the recovery in those patients one month after a course of treatment. Two case-control studies were respectively conducted, in which 161 patients and 161 healthy subjects as well as 41 uncured patients and 41 cured patients were randomly selected and matched for gender and age. Statistical analysis was performed using SPSS19. 0. Re-sults The two case-control studies showed that vitamin D deficiency was a risk factor for both Ct genital tract infection and poor efficacy, of which the adjusted ORs were 2. 281 (95％ CI: 1. 438, 3. 619) and 7. 266 (95％ CI: 2. 551, 21. 036). Among all subjects aged 20-39, male patients had lower 25-(OH)D level in serum than healthy men [(40. 10±17. 93) nmol/ L vs (53. 72±18. 00) nmol/ L, P< 0. 01] and fe-male patients also had lower 25-(OH)D level in serum than healthy women [(35. 71±19. 99) nmol/ L vs (45. 42±16. 08) nmol/ L, P<0. 01]. The levels of 25-(OH)D in uncured male and female patients were re-spectively lower than those in cured male and female patients [(30. 50±14. 53) nmol/ L vs (41. 32±17. 24) nmol/ L; (29. 47±16. 66) nmol/ L vs (41. 37±21. 03) nmol/ L; both P<0. 05]. Conclusion Vitamin D deficiency related to Ct infection in human genitourinary tract and poor prognosis. Lower serum vitamin D levels might increase the risk of Ct genitourinary tract infection and reduce the efficacy of treatment.

OBJECTIVE: To perform gene mutation analysis in a Chinese pedigree with dystrophic epidermolysis bullosa pruriginosa (DEB-Pr), and explore phetotype, genotype, and genotypes-phenotypes relationship of DEB-Pr. METHODS: Potential variants of the COL7A1 gene were detected by skin targeted sequencing panel and verified by Sanger sequencing. The pathogenicity of the variation was analyzed. RESULTS: Compound heterozygous variants, c.4128delT and c.8234G>A, were detected in the COL7A1 gene of the two patients. The c.4128delT(p.Pro1376fs) variant was derived from their mother and unreported previously. According to the American College of Medical Genetics and Genomics Standards and Guidelines, it was suggested to be a pathogenic mutation. The c.8234G>A(p.Arg2745Gln) variant was derived from their father, and possibly is a pathogenic variation. CONCLUSION In this study, the compound heterozygous variants of c.4128delT(p.Pro1376fs) and c.8234G>A(p.Arg2745Gln) of the COL7A1 gene probably underlies the disease in this patient and his sister. And our study expands the database on mutations of DEB-Pr.
Collagen Type VII/genetics* ; Epidermolysis Bullosa Dystrophica/genetics* ; Female ; Humans ; Male ; Mutation ; Pedigree ; Phenotype