Objective To investigate the effects of human papilloma virus 16 E6 (HPV16 E6) on endoplasmic reticulum (ER) stress-autophagic response in the cervical cancer C33A cells.Methods Polymerase chain reaction was used for detecting the integration of HPV DNA.The eukaryotic expression vector of HPV16 E6 was constructed and transfected via lipofectamine into C33A cells.Experimental cells were classified into 3 groups:pcDNA3.1--HPV16 E6 group,pcDNA 3.1-group and C33A group.Western blot was used to measure expression of protein of HPV16 E6,Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 in transfected cells.Results here was no HPV DNA integration in C33A cells that were confirmed as the intervention cells.Eukaryotic expression vector pcDNA3.1--HPV16 E6 was constructed successfully.The eukaryotic expression vector pcDNA3.1--HPV16 E6 significantly improved the expression of protein of HPV 16 E6 in C33A cells.The protein expression of Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 were significantly improved after transfection with vector pcDNA3.1 +-HPV16 E6 (P ＜ 0.05).Furthermore,LC3 Ⅱ protein level was reduced by treatment with ER stress inhibitor.Conclusion HPV16 E6 can improve autophagy through the ER stress pathway,and this response may play an important role in the process of HPV16 E6 inducing cervical cancer,providing one of the new strategies for gene therapy of cervical carcinoma.

Objective To evaluate whether an early change in 18F-fluorodeoxyglucose (18F-FDG) uptake can predict tumor response to epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) and prognosis in patients with non-small cell lung cancer (NSCLC). Methods From August 2009 to April 2015, 22 patients with NSCLC who were eligible to EGFR-TKI treatment were enrolled. PET-CT scan was performed before (baseline) and 1 month after EGFR-TKI administration. Up to 5 hottest single tumor lesions (no more than 2 per organ) were considered to be target lesions. Maximum standardized uptake values (SUVmax) were measured, and post-treatment percentage changes in SUVmax (ΔSUV%) were calculated. PET responses were classified using PET response criteria in solid tumors (PERCIST). Then conventional CT scan was performed every 2 months for follow-up. Kappa statistic was used to compare agreement between the RERCIST recommendations-based therapeutic response evaluation and those based on RECIST1.1 criteria. Fisher exact test was used to compare the probability of disease progression in the early metabolic response and non-response groups. Predictive accuracy of ΔSUV% with respect to response or non-progression at CT scan was evaluated by ROC analysis. Progression-free survival (PFS) was determined by Kaplan-Meier survival analysis, and between-group comparison was performed by log-rank test. Results After 1 month of EGFR-TKI treatment, 12 patients (55%) showed partial metabolic response (PMR), 6 (27%) had stable metabolic disease (SMD), and 4 (18%) had progressive metabolic disease (PMD). There was a moderate agreement(Kappa=0.506,P<0.05) between PET response at 1 month based on PERCIST recommendations and CT response at 3 months according to RECIST 1.1. Non-progression was significantly more frequent in patients with an early PMR (χ2=11.941, P=0.005). Progression had been confirmed later during therapy in all patients with PMD . By using ROC analysis, the area under the curve for prediction of response was 0.906 (95% CI, 0.766—1.000; P=0.002), corresponding to a sensitivity of 88.9% and specificity of 84.6% at a cut-off of 40.36% in ΔSUV%. Using a cut-off value of 25.84% in ΔSUV%, highΔSUV% group (ΔSUV% ≥ 25.84%) had significantly longer PFS than low ΔSUV% group (ΔSUV%<25.84%). Conclusion Early assessment of PET-CT at 1 month of EGFR-TKI treatment could be useful to predict tumor response and clinical outcome in patients with NSCLC.

Objective To evaluate the effects of turmeric volatile oil (TVO) combined with cisplatin on the proliferation and apoptosis of a human cutaneous squamous cell carcinoma cell line A431,and to explore their mechanisms.Methods Some cultured A431 cells at exponential growth phase were divided into several groups to be treated with 5,10,20,40 and 80 mg/L TVO,as well as high-glucose Dulbecco's modified Eagle's medium (DMEM) containing 1％ dimethyl sulfoxide (DMSO,control group),respectively.After 24-hour treatment,cell counting kit 8 (CCK8) assay was performed to estimate the proliferative activity of A431 cells in the above groups.Some other A431 cells were divided into 4 groups:control group treated with high-glucose DMEM containing 1％ DMSO,TVO group treated with 40 mg/LTVO,cisplatin group treated with 10 mg/L cisplatin,and TVO + cisplatin group treated with 40 mg/L TVO and 10 mg/L cisplatin.After 24-hour treatment,CCK8 assay was performed to estimate the cellular proliferative activity,inverted microscopy to observe changes in cell morphology,fluorescence microscopy to detect cell apoptosis after acridine orange (AO)/ethidium bromide (EB) double-staining,colorimetry to evaluate the activity of Caspase-3 and Caspase-9,and Western blot analysis to determine the protein expression of Caspase-3 and p-glycoprotein.Results After 24-hour treatment with 5,10,20,40 and 80 mg/L TVO,the cell proliferation rates were inhibited by (12.83 ± 6.4)％,(16.27 ± 11.4)％,(21.61 ± 9.1)％,(33.11 ± 2.0)％ and (46.00 ± 3.3)％ respectively,and the inhibition rates were all significantly higher in these groups than in the control group (4.03％ ± 1.4％,all P ＜ 0.05).The 50％ inhibitory concentration (IC50) of TVO at 24 hours was (61.66 ± 1.03) mg/L.Compared with the control group,the proliferation inhibition rates significantly increased in the TVO group,cisplatin group and TVO + cisplatin group (all P ＜ 0.05),suggesting that the combination of TVO and cisplatin showed synergistic inhibitory effects with a combination index of 1.366.Moreover,A431 cells turned round to different extents and became apoptotic in the TVO group and cisplatin group,and the TVO + cisplatin group showed obviously decreased number of cells and a large number of cell debris.The TVO + cisplatin group also showed significantly increased activity of Caspase-3 (1.520 ± 0.115) and Caspase-9 (2.760 ± 0.297) as well as protein expression of Caspase-3 (1.482 ± 0.016) compared with the TVO group (Caspase-3 activity:1.117 ± 0.095;Caspase-9 activity:1.259 ± 0.059;Caspase-3 protein expression:1.156 ± 0.006,all P ＜ 0.01) and cisplatin group (Caspase-3 activity:1.381 ± 0.089;Caspase-9 activity:1.829 ± 0.171;Caspase-3 protein expression:1.296 ± 0.021,all P ＜ 0.01),but significantly decreased p-glycoprotein expression (0.528 ± 0.014) compared with the TVO group (1.311 ± 0.011,P ＜ 0.01) and cisplatin group (1.169 ± 0.012,P ＜ 0.01).Conclusion TVO combined with cisplatin can synergistically inhibit the proliferation of A431 cells and induce cell apoptosis,which may be associated with activation of the caspase system and decreased expression of pglycoprotein.

Objective To understand the characteristics and relation of clinical stage and outcome of severe cases on hand,foot and mouth disease (HFMD) and to establish the evaluation method for understanding severity of this disease.Methods According to factors as geographical location,economic and epidemic levels,five provinces (Henan,Shandong,Yunnan,Zhejiang and Sichuan provinces) were selected.Reported severe cases of HFMD from the National Notifiable Diseases Reporting System were selected randomly in the five provinces.Basic epidemiological information,clinical data,and pathogen testing results in the involved hospitals were collected.Clinical stages on all the patients were decided in accordance with "the clinical expert consensus on diagnosis and treatment for severe case of enterovirus type 71 (EV71) infections (2011 edition)".Data were analyzed using SPSS software 18.0 and other epidemiological methods.Results A total of 657 severe HFMD cases were investigated,with 326 cases positive of EV71,accounting for 91.3％ (326/357) among all the laboratory-confirmed cases.Of the 657 cases,542 cases (82.5％,95％CI:79.4％-85.3％) were diagnosed as in stage 2 (with nervous system involvement),99 cases (15.1％,95％CI:12.4％-18.0％) in stage 3 (early phase of function failure on heart and lung),and 16 cases (2.4％,95％CI:1.4％-3.9％) were in stage 4 (function failure of heart and lung).11 cases (1.7％,95％CI:0.9％-3.0％) were with squeal when discharged from hospital with 8 cases (1.2％,95％CI:0.6％-2.3％) died.When comparing the proportions among stage 2,stage 3 and stage 4,significant differences were found between age groups (x2=22.632,P=0.012).The younger the patient was the lower the proportions of stage 2 and the more proportion of stage 3 appeared.When comparing the proportions of clinical stages among the five provinces,significant differences (x2=41.481,P =0.000) were noticed.Proportions of different clinical stages in gender,ethnicity,occupation,place of residence types and the type of pathogen appeared no significant differences,respectively.However,the proportions of squeal and death in stage 2,stage 3 and stage 4 showed significant differences (sequela:x2=12.960,P=0.001;Death:x 2=16.850,P=0.001),respectively.Conclusions The pcrccntage of clinical stages of severe HFMD patients related to the rate of squeal and death.Clinical staging can be used for assessing the clinical severity of complications and the effectiveness of treatment,of HFMD.

Objective: To observe the effect of Tai Chi combined with conventional western medicine therapy for elderly patients with type2 diabetes mellitus. Methods: According to randomized controlled design, 108 patients were divided into two groups ,with 54 in each group. Patients in control group were treated with conventional western medicine therapy and regular walk (three times per week), while the patients in the observation group were treated with conventional western medicine therapy and Tai Chi (three times per week), both groups were treated for 12 weeks. Before and after treatments, diabetes symptoms scale was used to assess disease conditions, FPG and HbA1c were detected separately by methods of HK (hexokinase) and High Performance Liquid Chromatography (HPLC), Diabetes mellitus Specific Quality of Life scale (DSQL) were used to assess the quality of life, Self-Efficacy for Managing Chronic Disease 6-Item Scale (SES6G) was used to assess patients’ self-management ability, and Geriatric Depression Scale was used to assess patients' depression emotions. Results: After treatments, the clinical symptoms scores (1.79 ± 1.21 vs. 2.86 ± 1.66, t=-3.401), the DSQL scores (50.40 ± 10.88 vs. 56.00 ± 10.85, t=-2.156), the GDS scores (5.63 ± 0.71 vs. 6.08 ±0.91, t=-2.515) of observation group were significantly lower than that of control group (P<0.01 or P<0.05), and the SES6G scores (7.54 ± 0.59 vs. 6.89 ± 0.39, t=-5.987) of observation group were significantly higher than that of control group (P<0.01); after treatments, the D-value of FPG (-0.97 ± 1.54 mmol/L vs. -0.28 ± 1.74 mmol/L, t=-2.092) was significantly higher than that of control group (P<0.05). Conclusions The Tai Chi combined with conventional western medicine therapy could alleviate clinical symptoms and negative emotions, improve disease management and quality of life, regulate blood glucose of elderly patients with type 2 diabetes mellitus.

Objective To investigate the use of hepatoprotective drugs in China in recent years, and to put forward related suggestions. Methods The outpatient prescription data of hepatoprotective drugs were collected from 85 hospitals in 6 cities of China from 2015 to 2019, and a real-world data analysis was performed to analyze the payment method, issuing department, drug category, and use of hepatoprotective drugs. Results A total of 1 113 575 prescriptions were extracted, involving 38 hepatoprotective drugs such as compound glycyrrhizin, polyene phosphatidylcholine, and bicyclol. Hepatoprotective drugs were mainly in tertiary hospitals, and the highest number of prescriptions containing hepatoprotective drugs were observed in department of infectious diseases, department of gastroenterology, and department of tuberculosis. Anti-inflammatory hepatoprotective drugs accounted for the highest proportion of all prescriptions, mainly compound glycyrrhizin, polyene phosphatidylcholine, and bicyclol. Of all prescriptions, 253 429 (22.76%) had the combination of multiple hepatoprotective drugs, with the highest number of 6 drugs, among which polyene phosphatidylcholine combined with bicyclol accounted for the highest proportion. Conclusion There are large quantities of hepatoprotective drugs used by outpatients in China. At present, the hepatoprotective drugs are clinically applied rationally, but there are still some problems to be solved, such as the combination of drugs.