OBJECTIVE Based on the G protein-coupled bile acid receptor(TGR5)/nucleotide binding oligomerization domain-like receptor 3(NLRP3) signaling pathway, to explore the mechanism of Huoxue Qingjieling in improving the inflammatory response of rats with nonalcoholic steatohepatitis(NASH). METHODS A total of 70 male SD rats were randomly divided into 5 groups, 20 rats in the control group, 20 rats in the model group, 10 rats in each of the atorvastatin positive drug group, the high-dose and low-dose groups of Huoxue Qingjieling. The control group was given normal feed, and the rest of the groups were given high-fat diet. Through model evaluation, it was determined that the NASH rat model was successfully established at the end of the 20th week. After successful modeling, the control group and the model group were given with normal saline by intragastric administration, the atorvastatin positive drug group, and the high and low dose groups of Huoxue Qingjieling were given corresponding drugs once a day for 4 weeks. At the end of the 24th week, the rats were killed, and the changes of body weight, wet liver weight and liver index was calculated. The serum was taken to test the triglyceride(TG), total cholesterol(TC), alanine aminotransferase(ALT), and aspartic acid aminotransferase(AST) levels by automatic biochemical analyzer. Pathological changes of liver tissues were observed by HE staining and oil red O staining. The expression levels of TGR5 and NLRP3 proteins were detected by Western blotting and immunofluorescence staining. ELISA detected the content of interleukin-18(IL-18) and interleukin-1β(IL-1β) in liver tissue. RESULTS Huoxue Qingjieling could significantly improve the general state of NASH rats. Every dose group could significantly reduce the body weight, liver wet weight and liver index of rats(P<0.01), and TG, TC content and ALT, AST activity levels of serum significantly decreased(P<0.01). The pathological results showed that Huoxue Qingjieling could significantly improve liver steatosis, inflammation and balloon-like. The expression of TGR5 protein was significantly increased(P<0.01), the expression of NLRP3 protein and the content of IL-18, IL-1β were significantly decreased(P<0.01, P<0.05). CONCLUSION Huoxue Qingjieling can significantly improve the state of NASH rats, inhibit liver steatosis and inflammation, and its mechanism may be closely related to the inhibition of TGR5/NLRP3 signaling pathway.

Objective To investigate the effect of miR-199a-5p on common bile duct ligation(BDL)-induced liver fibrosis in rats by regulating intestinal flora.Methods The 25 SD rats were randomly divided into five groups:the Sham group,the BDL group,the negative control adenovirus(NC adv)group,the miR-199a-5p adv group and the miR-199a-5p sponge adv group.The pathological changes of liver tissue and the degree of liver fibrosis were ob-served by HE,Masson and Sirius Red staining.The levels of aspartate aminotransferase(AST),alanine amin-otransferase(ALT),total bilirubin(TBIL)and direct bilirubin(DBIL)in serum of rats were determined by a fully automatic biochemical analyzer.The mRNA expression level of miR-199a-5p in liver tissue of rats was detec-ted by qRT-PCR.The protein expression levels of α-smooth muscle actin(α-SMA)and collagen type 1 alpha 1(COL1A1)in liver tissue of rats were detected by double immunofluorescence staining and Western blot experi-ment.Rat feces were collected for 16S rRNA high-throughput sequencing.Results The expression of miR-199a-5p was up-regulated in the liver tissue of BDL rats(P＜0.01).Compared with the NC adv group,the degree of liver injury and collagen deposition were relatively serious,the levels of AST,ALT,TBIL and DBIL in serum and the expression levels of α-SMA and COL1A1 in liver tissue increased in the miR-199a-5p adv group(all P＜0.05).However,the results of miR-199a-5p sponge adv intervention were opposite(all P＜0.05).The 16S rRNA sequencing results showed that rats treated with miR-199a-5p adv were characterized by increased diversity and richness of intestinal microbiota,changed composition of intestinal microbiota,while the results of miR-199a-5p sponge adv interfering with the bacterial community were opposite(all P＜0.05).Conclusion miR-199a-5p promotes liver fibrosis of BDL rats,and its mechanism may be related to regulating the diversity and abundance of intestinal microbiota.

Objective To investigate the effect of CXXC finger protein 4 (CXXC4) on the proliferation and apoptosis of pancreatic cancer PANC-1 cells.Methods The expression level of CXXC4 in pancreatic canc-er tissues and its relationship with prognosis and clinicopathological stage of the patients were analyzed in on-line databases.The qRT-PCR technique was used to detect the mRNA expression level of CXXC4 in human normal pancreatic ductal epithelial cell line (HPNE) and pancreatic cancer cell PANC-1,AsPC-1 and BxPC-3. si-NC,si-CXXC4,pcDNA3.1 and pCDNA3.1-CXXC4 were respectively transfected into PANC-1 cells.West-ern blot was conducted to detect the effectiveness of CXXC4 knockdown and overexpression.CCK-8,colony formation,EdU and immunofluorescence assays were conducted to analyze the effect of CXXC4 knockdown or overexpression on the proliferation and apoptosis of PANC-1 cells.The bioinformatic websites was used to predict the upstream microRNA (miRNA) of CXXC4.The Starbase database was adopted to analyze the cor-relation between miR-450b-5p and CXXC4 expression in pancreatic cancer tissues.Results The TCGA data-base results showed that the expression of CXXC4 in pancreatic cancer tissues was lowly expressed compared with in paracancerous pancreatic tissues (P<0.001),moreover which was associated with the overall survival and poor prognosis in the patients with pancreatic cancer (P<0.05).The GEPIA database analysis results showed that compared with stage Ⅰ pancreatic cancer,the CXXC4 expression in stage Ⅱ pancreatic cancer was decreased (P<0.05).Compared with HPNE cells,the CXXC4 expression in 3 kinds of pancreatic cancer cells was decreased (P<0.05).Compared with the si-NC group,the proliferation and colony formation ability of PANC-1 cells in the si-CXXC4 group were enhanced,the expressions of proliferation markers Ki67 and PC-NA were increased,and the expressions of apoptosis markers Bax,caspase-3 and caspase-9 were decreased;compared with the pcDNA3.1 group,the PANC-1 cells in the pcDNA3.1-CXXC4 group obtained the opposite results (all P<0.05).The bioinformatic websites predicted that miR-450b-5p was the upstream miRNA of CXXC4,CXXC4 in pancreatic cancer tissues was negatively correlated with the miR-450b-5p expression (r=-0.227) and miR-450b-5p in various mammalian species was highly conserved.Conclusion CXXC4 inhibits the proliferation of PANC-1 cells in pancreatic cancer and promotes theirs apoptosis.

Epidermal growth factor receptor (EGFR) is one of the most common targeted oncogenes in non-small cell lung cancer (NSCLC). The third-generation EGFR tyrosine kinase inhibitors (TKIs) have become the standard treatment for metastatic or recurrent NSCLC patients harboring EGFR positive or concomitant T790M mutations. However, the inevitable emergence of acquired resistance markedly limits their prolonged clinical benefits, although the third-generation EGFR TKIs have shown potent clinical outcomes in initial several months. This paper firstly reviews the characEpidermal growth factor receptor (EGFR) is one of the most common targeted oncogenes in non-small cell lung cancer (NSCLC). The third-generation EGFR tyrosine kinase inhibitors (TKIs) have become the standard treatment for metastatic or recurrent NSCLC patients harboring EGFR positive or concomitant T790M mutations. However, the inevitable emergence of acquired resistance markedly limits their prolonged clinical benefits, although the third-generation EGFR TKIs have shown potent clinical outcomes in initial several months. This paper firstly reviews the characteristics and clinical efficacy of the third-generation EGFR TKIs in the market or in the clinical development. Then this article summarizes the detailed mechanisms behind the acquired drug resistance of third-generation EGFR TKIs，and further expounds the current treatment strategies to overcome the resistance. Collectively, this review could provide more information for the development and clinical application of drugs targeting EGFR.

Objective:To investigate the relationship between plasma Golgi protein 73 (GP73) levels and the occurrence and development of non-alcoholic fatty liver disease (NAFLD), and to establish a diagnostic model based on this combination with lipid metabolism indicators to clarify its diagnostic efficacy and clinical application value for NAFLD.Methods:225 cases with NAFLD [diagnosed by ultrasound, transient elastography (FibroScan502) and liver biopsy (some patients)] and 108 healthy controls were selected from the Department of Hepatology and Physical Examination Center of Integrated Traditional Chinese and Western Medicine, The Third Hospital of Hebei Medical University. Clinical data, routine peripheral blood and serum biochemical test results were collected. The plasma GP73 level was detected by enzyme-linked immunosorbent assay. SPSS 21.0 statistical software was used for statistical analysis. Binary logistic regression model was used to calculate the NAFLD diagnostic model. Receiver operating characteristic curve was used to evaluate the NAFLD constructed model diagnostic efficacy.Results:NAFLD incidence was significantly reduced in younger age group, mostly in young and middle-aged male. However, the NAFLD incidence was increased with increasing age in female. The analysis of age ratio composition showed that the average age for NAFLD onset was 20 ~ 50 years old, and the incidence rate was as high as 47% in among 30 ~ 39 years old, but the incidence rate was significantly decreased in over 60 years old (4.00%). GP73 was an independent risk factor for the occurrence and development of NAFLD. The diagnostic models of GBT, GB and GT were established by GP73 (G) combined with body mass index (BMI, B) and serum triglyceride (TG, T), and the results showed that the areas under the curves of GBT, GB and GT models were 0.969, 0.937 and 0.909, respectively. The sensitivity and the specificity were 84.90%, 77.80% and 84.00%, and 95.40%, 95.40% and 82.40%, respectively, P < 0.05. The GBT model had efficacy of best diagnostic performance. Conclusion:NAFLD is more common in young and middle-aged male, but with advanced age, the incidence of female patients gradually increases. Plasma GP73 levels are related to the occurrence and development of NAFLD. The GBT model can be used as a new model for non-invasive diagnosis and one of the indicators for clinical evaluation of diagnostic efficacy of NAFLD.

Objective:To clarify the clinical efficacy of Yiqi Huoxue recipe in the treatment of liver fibrosis of chronic viral hepatitis.Methods:An open, positive-drug, parallel-controlled study method was applied. A total of 207 cases of liver fibrosis with chronic hepatitis B and C diagnosed with liver biopsy and transient elastography were selected. According to the principle of syndrome differentiation in traditional Chinese medicine, self-made Yiqi Huoxue recipe ( n = 127) and Fuzheng Huayu capsule ( n = 80) were used for the treatment course of 24-48 weeks. Change score of TCM symptom, liver biochemistry, liver stiffness measurement (LSM), and noninvasive liver fibrosis index [aspartate transaminase to platelet ratio index (APRI), and fibrosis-4 score (FIB-4)] were compared between the two groups to evaluate the therapeutic effect of Yiqi Huoxue recipe on liver fibrosis. Results:Yiqi Huoxue recipe group and Fuzheng Huayu capsule group baseline LSM, APRI and FIB-4 was compared, and there was no statistically significant difference between them ( P > 0.05). Yiqi Huoxue recipe and Fuzheng Huayu capsule received patients had improved symptom scores to a certain extent. Hepatic facies, discomfort over liver area, and soreness and weakness of waist and knees ( P < 0.05) was significantly improved in Yiqi Huoxue recipe than Fuzheng Huayu capsule. Liver biochemical indicators (ALT, AST, GGT, ALP) had gradually relapsed with the extension of treatment duration and the normalization rate between the two groups after 24 to 48 weeks had reached 100% vs. 100%, 100% vs. 93.8%, 96.8% vs. 92.3% and 87.5% vs. 81.8%. After 12 weeks of treatment, APRI values ??of both groups had significantly reduced, and after 48 weeks of treatment, LSM values of both groups had significantly improved. Moreover, Yiqi Huoxue recipe FIB-4 score was significantly improved after 48 weeks of treatment, and the difference was statistically significant compared to Fuzheng Huayu capsule group ( P < 0.05). After treatment, LSM, APRI, and FIB-4 total effectiveness in the two groups were 80.0% vs. 63.6%, P = 0.046; 68.4% vs. 52.0%, P = 0.052; 68.4% vs. 62.0%, P = 0.437, respectively. LSM total effectiveness was significantly higher in Yiqi Huoxue recipe treated group than Fuzheng Huayu capsule group. Conclusion:Traditional Chinese medicine Yiqi Huoxue decoction can be used as an optimal treatment for liver fibrosis of chronic viral hepatitis.

Objective:To study the bacterial pathogen, the optimal plan of antibacterial drugs and the prognostic factors in patients with liver cirrhosis combined with bacterial pneumonia.Methods:Data of 324 cases with liver cirrhosis from the Department of Traditional and Western Medical Hepatology, the Third Hospital of Hebei Medical University were collected, including 217 cases of bacterial pneumonia. Baseline characteristics of the patients, factors affecting the efficacy of antibacterial treatment and prognosis were compared and analyzed. Logistic regression analysis was used to screen and predict the antibacterial efficacy indicators and a prediction model was established. Receiver operating characteristic curve was used to evaluate the value of the established model and Child-Turcortte-Pugh, model for end-stage liver disease, and model for end-stage liver disease combined with serum sodium concentration predict the therapeutic efficacy.Results:Chronic HBV and HCV infections were the main causes of cirrhosis, followed by cryptogenic cirrhosis and alcoholic cirrhosis. Diabetes, cardio-cerebrovascular and chronic obstructive pulmonary disease were susceptible factors for bacterial pneumonia. As infection occurred, the ratio of neutrophils to lymphocytes, serum C-reactive protein, procalcitonin, alanine aminotransferase, and total bilirubin levels had increased significantly. The results of pathogenic analysis showed that the top three pathogenic bacteria were Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus aureus. The resistance rate of Klebsiella pneumoniae to ceftriaxone was 50.0%, and that of ceftazidime, cefepime, and cefoperazone sulbactam were 27.8%. Imipenem and piperacillin tazobactam containing β-lactamase inhibitors were the most effective antibacterial therapies. Regression analysis showed that age, procalcitonin, and albumin was significantly correlated with antibacterial effects. The PAA model was established and had predicted the efficacy of Child-Turcortte-Pugh, model for end-stage liver disease, and model for end-stage liver disease combined with serum sodium. The specificity and sensitivity of the PAA was confirmed to be 94.12% and 93.62%, which was significantly higher than other models.Conclusion:The main common pathogenic bacterium of cirrhosis combined with bacterial pneumonia is Klebsiella pneumonia (G-bacilli). In addition, gram positive cocci (Staphylococcus aureus) and other are also visible. The elderly, diabetics and patients using hormones are prone to secondary fungal infections. Age, procalcitonin and serum albumin can accurately predict the antibacterial effect, guide clinical treatment and judge the prognosis of the established PAA model.

Direct-acting antiviral agents (DAAs) are the main antiviral therapeutics for hepatitis C virus-related decompensated stage cirrhosis. DAAs of NS3/4A protease inhibitors use is not recommended for patients with decompensated cirrhosis due to characteristics of DAAs metabolism in liver. The recent guidelines have recommended sofosbuvir (SOF)-based plan including pan-genotype plan of sofosbuvir(SOF)/velpatasvir (VEL), sofosbuvir combined with daclatasvir (DCV), genotype 1,4,5,6 specific plan of sofosbuvir (SOF) / ledipasvir (LDV) for 24 weeks or above in combination with ribavirin for 12 weeks because NS5B and NS5A inhibitors has no obvious effect on CYP450 enzyme system and achievement of sustained virological response (SVR) rates at 12/24 weeks is achievable in 88% ~ 100%, and liver reserve function improves in 42% ~ 53% of patients. Furthermore, approximately 15.5% ~ 49% of patients waiting for liver transplantation after treatment with DAAs do not require liver transplantation for short-term and 10.3% ~19.2% of patients receiving SOF/LDV, and SOF combined with DCV not needed liver transplantation. Thus, the clinical application of DAAs provides a safe and reliable antiviral treatment plan for hepatitis C virus-related decompensated stage cirrhosis.

Objective: To explore the clinical value of plasma heme oxygenase 1(HO-1) in the development of non-alcoholic fatty liver disease(NAFLD). Methods: Patients with NAFLD were selected from the Physical examination center and the Department of Traditional and Western Medical Hepatology of Third Hospital of Hebei Medical University. A combination of ultrasound and liver elastography was used to screen NAFLD patients and healthy persons. General clinical characteristics, peripheral blood cell count and liver biochemical test results were collected synchronously, plasma samples were retained, and plasma HO-1 level was detected by enzyme-linked immunosorbent assay. SPSS21.0 statistical software was used for statistical analysis, multivariate logistic regression analyses was used to analyse the independent risk factors affecting the incidence and progression of NAFLD. The diagnostic efficacy of indicators related to development of NAFLD was assessed by the receiver operating characteristic curve(ROC). Results: A total of 328 patients with NAFLD and 113 healthy controls were included. According to the liver biochemical results, the NAFLD group was divided into 148 patients with normal liver enzymes and 180 patients with abnormal liver enzymes. The level of HO-1 in the three groups was 9.09 ± 2.19, 14.38 ± 2.63, 17.00 ± 3.30 ng/ml, and was increased respectively of healthy controls, patients with normal liver enzymes and patients with abnormal liver enzymes. Analyzing plasma HO-1 levels of components associated with metabolic disorders suggests that components without metabolic syndrome(9.83 ± 3.21) < components with 1 metabolic syndrome(13.59 ± 3.72) < components with 2 or more metabolic syndrome(16.09 ± 3.41), P < 0.001. The results of HO-1 level stratification analysis showed that WBC, ALT, AST, GGT, TG increased as HO-1 level increased, and the pairwise difference was statistically significant (P < 0.001). The WBC count of NAFLD is significantly higher than healthy group(6.79 ± 1.62 vs 5.68 ± 1.36, P < 0.001). The univariate and multivariate regression analyses of all the subjects showed that HO-1, TG and BMI were prognostic factors for the occurrence of NAFLD and HO-1, TC, GLU were prognostic factors for the progression of NAFLD, P < 0.05. The ROC analysis showed that HO-1 was reliable markers for predicting the occurrence and progression of NALFD, the sensitivity and specificity were respectively 85.10%, 92.90% and 38.33%, 95.27%. Conclusion Plasma HO-1 can predict the occurrence and progression of NAFLD and is expected to be a novel molecular diagnostic marker for NAFLD and NASH.

Objective To investigate the vitamin B 12 status of vegetarians in Shanghai.Methods A total of 282 adult vegetarians and 282 omnivores matching by gender and age were recruited in Shanghai.Their dietary intakes were collected.The serum concentrations of vitamin B12,folate and homocysteine were tested.The red blood cell,hematocrit value,mean corpuscular volume and mean erythrocyte width were also examined.Results The daily average intake of dietary vitamin B12 was (0.46± 1.01) μg/d in vegetarians and only (0.1±0.46) μg/d in vegans,which was lower than that of omnivores [(3.91±6.92) μg/d,F=50.57,P＜0.01].137 omnivores and 274 vegetarians had less dietary vitamin B12 level than recommended nutrient intake (RNI) and the difference was statistically significant (x2 =114.77,P＜ 0.01).54.26％ of vegetarians,68.92％ vegans,49.04％ ovo-lacto vegetarians and 15.60％ omnivores had hyperhomocysteinemia and the differences between vegetarians and omnivores were statistically significant (all P＜0.01).After adjusting the confounding factors the hematocrit value was higher in vegetarians,vegans and ovo-lacto vegetarians than in omnivores (27.42％± 18.32％,28.73％± 18.19％,26.95％± 18.38％ vs.8.96％± 16.59％,P＜0.01).Vegans had lower red blood cell counts and higher hematocrit value and mean corpuscular volume than omnivores.Conclusion Vitamin B12 deficiency combined with an elevated level of homocysteine and red blood cell volume growth are common but serious issue in vegetarians,especially in vegans.