Polyphyllin Ⅰ(PPⅠ)and polyphyllin Ⅱ(PⅡ)are the main active substances in the Paris polyphylla.However,liver toxicity of these compounds has impeded their clinical application and the potential hepatotoxicity mechanisms remain to be elucidated.In this work,we found that PPⅠ and PⅡ exposure could induce significant hepatotoxicity in human liver cell line L-02 and zebrafish in a dose-dependent manner.The results of the proteomic analysis in L-02 cells and transcriptome in zebrafish indicated that the hepa-totoxicity of PPⅡ and PⅡwas associated with the cholesterol biosynthetic pathway disorders,which were alleviated by the cholesterol biosynthesis inhibitor lovastatin.Additionally,3-hydroxy-3-methy-lglutaryl CoA reductase(HMGCR)and squalene epoxidase(SQLE),the two rate-limiting enzymes in the choles-terol synthesis,selected as the potential targets,were confirmed by the molecular docking,the over-expression,and knockdown of HMGCR or SQLE with siRNA.Finally,the pull-down and surface plasmon resonance technology revealed that PPⅠ could directly bind with SQLE but not with HMGCR.Collectively,these data demonstrated that PPⅠ-induced hepatotoxicity resulted from the direct binding with SQLE protein and impaired the sterol-regulatory element binding protein 2/HMGCR/SQLE/lanosterol synthase pathways,thus disturbing the cholesterol biosynthesis pathway.The findings of this research can contribute to a better understanding of the key role of SQLE as a potential target in drug-induced hepatotoxicity and provide a therapeutic strategy for the prevention of drug toxic effects with similar structures in the future.

Objective:Inhaler satisfaction is an important factor affecting inhaler adherence and the efficacy of inhalers in chronic airway diseases.Using a scientific and effective method to assess patients'satisfaction with inhalers is of great significance for improving clinical outcomes.The Feeling of Satisfaction with Inhaler-10(FSI-10)is specifically designed to assess patients'inhaler satisfaction in chronic airway diseases,but the application research on this scale is not available in China.This study aims to evaluate the reliability and validity of the Chinese version of FSI-10,describe the current status of inhaler satisfaction and discuss the associated variables in Chinese patients with chronic airway disease. Methods:Based on the English version of FSI-10,items of the Chinese version of FSI-10 were determined after forward-backward translation and cultural adaption.Totally,322 patients with chronic obstructive pulmonary disease(COPD)and asthma were enrolled from the Second Xiangya Hospital of Central South University from June to October 2022.We collected associated clinical variables and inhaler satisfaction using the Chinese version of FSI-10.The content validity of the scale was expressed by content validity index(CVI)and the construct validity was analyzed by exploratory factor analysis(EFA).The reliability of the scale was expressed by Cronbach's α coefficient,the split-half reliability and test-retest reliability.A multivariate logistic regression was conducted to examine variables related to inhaler satisfaction. Results:The reliability and validity analysis showed that the CVI was 0.983.One factor was extracted from the Chinese version of FSI-10 and the cumulative variance contribution rate was 73.114%.The Cronbach's α of the scale was 0.913,the Guttman's half-reliability coefficient was 0.905,and the test-retest reliability was 0.727(P<0.001).In addition,the total score of the scale for patients was 38.92±4.26 points and the proportion of high satisfaction(the score of FSI-10≥40)in patients with COPD was significantly lower than that in asthma patients(71.3%vs 87.9%,P<0.01).Older age(age≥70 years)was a risk factor of lower inhaler satisfaction and asthma diagnosis was a protective factor. Conclusion:The Chinese version of FSI-10 has good reliability and validity in patients with COPD and asthma,which may be further promoted and applied in patients with chronic airway disease in China.Doctors should regularly evaluate the inhaler satisfaction of patients with chronic airway diseases,especially for those elder or with severe symptoms and a long course of illness.

Objective:To explore the application and clinical significance of the cancer genome atlas (TCGA) molecular classification in endometrial cancer (EC).Methods:Sixty-six EC patients collected from December 2018 to March 2021 from Peking University People′s Hospital were categorized into four subgroups based on TCGA molecular classification tested by next generation sequencing. The correlation among four molecular subgroups and the clinical-pathological features including prognosis were analyzed.Results:(1) Clinical and pathological features: median age at diagnosis was 56 years (range: 24-78 years). The cases were distributed as follows: 3 (5%) cases DNA polymerase epsilon (POLE) ultra-mutated, 11 (17%) cases high microsatellite instability (MSI-H) including 2 Lynch syndrome, 42 (64%) cases low copy-number (CN-L) and 10 (15%) cases high copy-number (CN-H). There were significant differences among four subtypes in the combination of other tumors, tumor family history, surgical method, International Federation of Gynecology and Obstetrics (FIGO, 2009) stage, depth of muscle invasion and lymph vascular space invasion (all P<0.05). The proportions of patients in CN-H subgroup with advanced FIGO stage (stage Ⅲ-Ⅳ), deep muscle invasion and positive lymph-vascular space invasion were significantly increased. There were no significant differences in age, menopausal status, body mass index, metabolic syndrome-related complications, preoperative serum CA 125 and human epididymis protein 4 levels, tumor size, pathological grade (only endometrioid cancer), and lymph node metastasis among the 4 TCGA molecular types (all P>0.05). (2) Immuno-related molecular analysis: among 66 EC patients, 27 patients underwent immunohistochemical analysis of programmed cell death 1 ligand 1 (PD-L1) protein, and 28 patients underwent tumor mutation burden (TMB) detection. POLE and MSI-H subgroups contained TMB than those in CN-L and CN-H ( P<0.05).(3) Prognosis: the median follow-up time was 10 months (range: 0-28 months). The progression-free survival rate of TCGA molecular types were 100% (POLE ultra-mutated), 100% (MSI-H), 98% (CN-L), and 80% (CN-H) respectively and had significant differences ( P=0.034). The overall survival were 100% (POLE ultra-mutated), 100% (MSI-H), 98% (CN-L), and 90% (CN-H) respectively, but there were not statistically significant difference ( P=0.361). POLE ultra-mutated and MSI-H subgroups had the best survival, while CN-H had the worst. Conclusion:TCGA molecular classification has feasibility and clinical value in clinical application of EC, which is helpful to identify the prognosis of patients.

To investigate the hypolipidemic effects of gypenosides granules and its combination with lipitor, a model of hyperlipidaemia C57BL/6J mice was established by high-fat diet feeding for 4 weeks. The mice were randomly divided into blank group, model group, lipitor group(10 mg/kg of lipitor), low dose group(90 mg/kg of gypenosides granules), medium dose group(120 mg/kg of gypenosides granules), high dose group(180 mg/kg of gypenosides granules)and the combination group(180 mg/kg of gypenosides granules and 10 mg/kg of lipitor). After 4 weeks of continuous administration, the contents of serum lipid indexes, serum ALT, AST and apolipoprotein B(ApoB)were measured. The liver tissues of mice were observed by H&E staining. The expression levels of key factors involved in hepatic cholesterol metabolism were observed by RT-PCR and Western blot methods, such as adenosine triphosphate combined box transporter A1(ABCA1), liver X receptor(LXRα), cholesterol 7 alpha hydroxylase(CYP7A1)and type BΙ scavenger receptor(SR-BΙ). The results revealed that gypenosides granules significantly decreased the mice body weight, total abdominal fat area and the level of serum total cholesterol(TC). The combination group showed a more significant reduction in TC level than the other administration groups. Moreover, gypenosides granules treatment remarkably increased the protein expression of ABCA1 and up-regulated the mRNA expression of ABCA1, CYP7A1 and SR-BI. The above results suggest that gypenosides granules can significantly reduce blood lipid contents, and the combination therapy with lipitor show better the lipid-lowering effect. Meanwhile, gypenosides granules can decrease the level of serum transaminase. Preliminary exploration suggests the lipid-lowering mechanism of gypenosides granules may be involved in cholesterol reversal to regulate the level of TC.

Objective To evaluate the clinical outcomes and feasibility of multi-modality adjuvant chemotherapy and radiation, which was conducted as postoperative chemotherapy, radiation, and consolidation chemotherapy (CRC) mode for the treatment of advanced endometrial cancer. Methods A retrospective analysis of 124 patients with International Federation of Gynecology and Obstetrics (FIGO) stages Ⅲ and Ⅳ endometrial cancer from Jan. 2004 to Oct. 2012 was conducted in Peking University People′s Hospital and Beijing Obstetrics and Gynecology Hospital. Inclusion criteria were comprehensive staging procedure including hysterectomy, bilateral salpingo-oophorectomy, and (or) selective pelvic aortic lymphadenectomy, and treatment with adjuvant chemotherapy and (or) radiation. The average age of these patients was (55.9 ± 8.4) years old (range from 23 to 79 years old). According to different postoperative adjuvant treatment modes, the patients were divided into CRC group, chemotherapy-radiotherapy (CR) group and single chemotherapy (C) group. The survival and side effects of the three groups were compared. Results (1) One hundred and twenty-four patients with advanced stage endometrial cancer were identified and received postoperative adjuvant therapies.Sixty-one (49.2％, 61/124) cases of them received postoperative CRC fashion, 19 (15.3％, 19/124) received postoperative CR and 44 (35.5％, 44/124) cases received C. The age, stage, grade and type of surgery of the three groups were not significantly different (all P>0.05);while, the pathology, chemotherapy cycles and chemotherapy regimens differed significantly (all P<0.05). (2) The progression-free survivals (PFS) of the patients with CRC, CR, and C group were (121±7), (68± 15), and (100±11) months, respectively. The 3-year PFS rates were 87.9％, 43.7％, and 61.4％, respectively. The 5-year PFS rates were 82.2％, 36.4％, and 61.4％, respectively. The above indicators were significantly higher in the CRC group than in the CR group (all P<0.01), and there was no difference between the CRC group and the C group (P=0.037). The overall survival (OS) of patients with CRC, CR, and C group were (128 ± 6), (80 ± 12), and (99 ± 10) months, respectively. The 3-year OS rates were 87.8％, 72.4％, and 67.1％, the 5-year OS rate were 84.2％, 54.3％, and 64.1％, respectively. The above indicators were significantly higher in the CRC group than those in the CR group and C group (all P<0.01). (3) There was no difference in the frequency of adverse effects either chemotherapy, such as severe bone suppression or radiotherapy;hepatotoxicity,blood transfusion, dose modifications;or cycle delays between the CRC, CR and C group (all P>0.05). (4) In the univariate analysis shown that, stage, the fashion of postoperative adjuvant therapy and type of surgery were risk factors for tumor progression in patients with advanced endometrial cancer (P<0.05). After adjusted for FIGO stage and type of surgery, the tumor progression hazard ratio (HR) was 3.931 (95％CI:1.734-8.914, P=0.001) for the CR group and 2.188 (95％CI:1.010-4.741, P=0.047) for the C group, compared to the CRC group. Conclusion Sequential CRC delivered in a"sandwich"fashion for the treatment in advanced endometrial cancer could significantly improve the 3-year and 5-year OS rates and have a similar adverse effect profile compared with other sequencing modalities.

To investigate the therapeutic effects of triterpenoids from Cyclocarya paliurus on non-alcoholic fatty liver disease (NAFLD),the model of NAFLD in HepG2 cells was induced by free fatty acids (FFAs). Cytotoxicity of the triterpenoids from C. paliurus was determined by MTT method,and the effects of triterpenoids without cytotoxicity on intracellular triglyceride (TG)and superoxide dismutase (SOD)were detected by the kits. Data indicated that compound 4 [2α,3α,23-trihydroxy-12,20 (30)-dien-28-ursolic acid,TUA]had hypolipidemic and antioxidant activities. After being treated with TUA and FFAs for 24 h,the intracellular lipid content was observed using Oil Red O staining,and intracellular TG,malondialdehyde (MDA ),SOD and reactive oxygen species (ROS)levels were determined by the assay kits. The protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2),heme oxygenase-1 (HO-1 )and NAD (P)H quinone oxidoreductase 1 (NQO-1 )were measured by Western blot. The results showed that TUA significantly increased SOD activity,and decreased intracellular TG, ROS and MDA levels in FFAs-induced HepG2 cells. Moreover,TUA dramatically improved Nrf2,NQO-1 ,and HO-1 expression. However,the dramatic increase in TG,ROS,MDA levels and the reduction in SOD,NQO-1 and HO-1 expression following Nrf2 inhibitor brusatol treatment were observed. In conclusion,these results suggest that TUA has the therapeutic effect on NAFLD which may be associated with Nrf2 activation.

Objective To compare and study the two kinds of quality control methodologies related to intelligent quality management system ( iQM) and traditional quality control , and the quality control performance of iQM equivalent to traditional quality control were evaluated , ensuring the accuracy of the results of blood gas testing.Methods Beijing Chaoyang Hospital of Capital Medical University , Beijing Tiantan Hospital of Capital Medical University , Shanghai Longhua Hospital of Shanghai University of Chinese Medicine, and Sichuan Provincial People′s Hospital, these 4 medical institutions were selected to implement this study.During the period from June 2016 to December 2016, in the routine detection of total 3 712 specimen, the iQM and traditional quality control modes were used simultaneously to calculate the mean values of all blood gas parameters quality controls , SD, CV (%) and Sigma values, to evaluate the quality control performance and difference of the two quality control modes .Results During the process of testing blood gas samples from 3 712 specimen in 4 hospitals, iQM process control solution ( PCS) A, B, C ran 1 089, 7 678 and 154 quality control samples respectively , and 732 external quality control samples were run by traditional quality control mode .Considering the most sensitive parameters of blood gas testing pO 2, iQM PCS A, B, C′s Sigma value are higher than 8, however, the traditional quality control′s Sigma value are less than 6; For parameters pCO2, pO2and Na+, there exists significant difference between two quality control methods (P＝0.004 8,P＝0.000 1,P＝0.004 4,P＜0.01), other parameters pH, K+, Ca ++, Glu, Lac and Hct, there exists no significant difference between two quality control methods (P＝0.250 6, P＝0.062 3,P＝0.034 0,P＝0.346 9,P＝0.186 3,P＝0.823 1,P＞0.01).Totally 22 errors detected by iQM, includes 14 micro-clots and 8 interferences samples, which were not detected by traditional quality control .Conclusions The error in blood gas analysis mainly comes from the pre-analytical phase.iQM enhanced specimen inspection capabilities and make up for the inability of traditional quality control to monitor the quality of specimens , enabling full-scale, real-time, and dynamic monitoring of each specimen , powerful error detection capabilities , and automatic error correction capabilities . Besides, automatic documentation saves staff much time.The system can effectively ensure the accuracy of blood gas test results, meet the quality requirements of related laws and regulations and related industry standards , and also can meet the clinical intended use , providing new ideas for POCT quality management and improvement.

Necrosis is a form of cell death, which is related to various serious diseases such as cardiovascular disease, cancer, and neurodegeneration. Necrosis-avid agents (NAAs) selectively accumulated in the necrotic tissues can be used for imaging and/or therapy of related diseases. The aim of this study was to preliminarily investigate necrosis avidity of I-evans blue (I-EB) and its mechanism. The biodistribution of I-EB at 24 h after intravenous administration showed that the radioactivity ratio of necrotic to viable tissue was 3.41 in the liver and 11.82 in the muscle as determined by counting in model rats. Autoradiography and histological staining displayed preferential uptake of I-EB in necrotic tissues. nuclear extracts from necrotic cells exhibited 82.3% of the uptake in nuclei at 15 min, as well as 79.2% of the uptake at 2 h after I-EB incubation. The DNA binding study demonstrated that evans blue (EB) has strong binding affinity with calf-thymus DNA (CT-DNA) (=5.08×10 L/(mol/L)). Furthermore, the accumulation of I-EB in necrotic muscle was efficiently blocked by an excess amount of unlabeled EB. In conclusion, I-EB can not only detect necrosis by binding the DNA released from necrotic cells, but also image necrotic tissues generated from the disease clinically.

To investigate the effect and possible mechanisms of triterpenic acid-enriched fraction from Cyclocarya paliurus(TAE)on glucagon secretion in insulin-resistance pancreatic α cells, the model of insulin resistance in αTC1-6 cells was induced by long term exposure to high glucose. Experimental groups were divided as follow: control(5. 5 mmol/L glucose), model(25 mmol/L), TAE(1, 5, 10 μg/mL), and TAE(10 μg/mL)plus wortmannin(10 nmol/L)group. The supernatant and lysate of treated cells were collected to determine glucagon secretion by ELISA kit. The mRNA and protein abundance of IRS-1, PI3K and Akt were measured by qPCR and Western blot analysis. Results showed that TAE could not only significantly reduce glucagon secretion induce by high glucose in a dose-dependent manner, but also remarkably increased the mRNA and protein abundance of IRS-1, PI3K and Akt in αTC1-6 cells. However, these effects of TAE were reversed by PI3K inhibitor wortmannin. In conclusion, it suggested that TAE could improve the insulin resistance induced by high glucose in pancreatic α cells which may be related with the activation of IRS-1/PI3K/Akt signaling pathway.

The purpose of this study was to investigate the hypoglycemic effect and the safety of medicinal formula composed of Cyclocarya paliurus and Mulberry leaves. An experimental diabetic rat model was established by high energy diet plus small dosage of auoxan(ALX). At the same time, each group rats were given distilled water(blank and model), metformin(Met), Cyclocarya paliurus aqueous extract(CP), Mulberry leaves aqueous extract(ML)and the different proportions of aqueous extract mixtures of Cyclocarya paliurus and Mulberry leaves(CM1, CM2 and CM3), respectively. Fasting blood glucose(FBG), OGTT, TC, TG, LDL-C, HDL-C, insulin and liver and kidney function related index were gauged to evaluate the hypoglycemic effect and the safety of samples. The results showed that FBG level of the rats in CM1, CM2 and CM3 groups decreased 21. 64%, 16% and 12. 55%, respectively, comparing with that of model group. Moreover, FBG, glucose tolerance and pancreatic tissue morphology were remarkably improved in CM1 group. TC and LDL-C levels of rats in ML and CM3 groups decreased significantly compared with the those of Model group(P< 0. 05), which showed ML and CM3 were beneficial to regulate the blood lipid level in diabetic rats. Furthermore, all the administration groups had no adverse effect on liver function index. The down regulation of kidney function index of CP, ML, CM1, CM2 and CM3 groups comparing with model group indicated that Cyclocarya paliurus and Mulberry leaves could alleviate the injury of liver and kidney. Our results demonstrated that the medicinal formula composed of Cyclocarya paliurus and Mulberry leaves were favorable to reduce blood glucose and can regulate lipid metabolism without liver and kidney toxicity.