In patients with liver disease such as viral hepatitis and liver cirrhosis,renal injury and renal insufficiency can be generally classified as acute kidney injury( AKI),chronic kidney disease,and acute- on- chronic nephropathy. AKI can be classified as stage 1( risk stage),stage 2( injury stage),and stage 3( failure stage). Traditionally hepatorenal syndrome is classified as types Ⅰ and Ⅱ,and in recent years,type Ⅲ hepatorenal syndrome with organic renal injury has been proposed. Hepatorenal disorder( HRD) is used to describe any renal disease which occurs in patients with liver cirrhosis. At present,sensitive and accurate biochemical parameters used to evaluate renal function in patients with liver disease in clinical practice include estimated glomerular filtration rate,increase in serum creatinine within unit time,and serum cystatin C level,and urinary microalbumin level also plays an important role in the early diagnosis of nephropathy. Causes of liver disease,severity,complications including infection,nutritional status,therapeutic drugs,and underlying nephropathy may be associated with renal injury and renal insufficiency in patients with liver disease and should be differentiated.

Objective To investigate the effect of protein acetylation in host cells on the replication of hepatitis B virus( HBV) in hepatocytes,since HBV infection greatly threatens human health and the acetylation of encoding proteins in infected cells plays an important role in HBV replication and infection. Methods The deacetylase inhibitors trichostatin A( TSA) and nicotinamide( NAM) were used to stimulate HBV replication in Hep G2. 2. 15 and Hep AD38 cells,and the HBV replication markers were measured. The pan- acetylysin protein and Ac- H3 were examined by Western Blot. Results The stimulation of cells with TSA and NAM increased the overall acetylation level of proteins in cells,and the acetylation level increased in a time- and dose- dependent manner. In the Hep G2. 2. 15 and Hep AD38 cells,stimulation with TSA and NAM reduced HBs Ag level in the supernatant of cell culture and increased HBV DNA level in a time- and dose- dependent manner,while HBe Ag in the supernatant of cell culture and DNA in cells did not change significantly. Conclusion Acetylation of host proteins may be involved in and affect HBV replication in cells,and further analysis and determination of host proteins whose acetylation affects HBV replication in cells help to learn more about the regulation of HBV replication and provide new thoughts for the development of specific antiviral strategies.

Objective To investigate the clinical effect of Dahuang Zhechong capsules combined with entecavir in the treatment of chronic hepatitis B( CHB) patients with liver fibrosis. Methods A total of 100 CHB patients with liver fibrosis who visited or were hospitalized in Shenzhen Hospital of Peking University from October 2014 to January 2016 were enrolled and randomly divided into Western medicine group and combined treatment group,with 50 patients in each group. The patients in the Western medicine group were given entecavir,and those in the combined treatment group were given Dahuang Zhechong capsules in addition to entecavir. The course of the treatment was 48 weeks.The changes in liver function,HBV DNA load,four serum parameters of liver fibrosis,liver stiffness,and aspartate aminotransferase- to-platelet ratio index( APRI) / Fibro Index were observed in both groups. The t- test was used for comparison of continuous data between groups,and the chi- square test was used for comparison of categorical data between groups. Results Both groups showed significant reductions in serum levels of aspartate aminotransferase( AST),alanine aminotransferase( ALT),and HBV DNA load after 48 weeks of treatment( all P < 0. 05). After treatment,the four serum parameters of liver fibrosis all returned to normal after treatment,and the serum levels of hyaluronic acid,type Ⅲ precollagen,and type IV collagen showed significant differences between the two groups( all P < 0. 05),and the portal vein diameter,thickness of the spleen,liver stiffness and APRI / Fibro Index also showed significant differences between the two groups( both P < 0. 05). The combined treatment group had a significantly higher overall response rate than the Western medicine group( 92. 0%vs 72. 0%,χ2= 6. 775,P = 0. 009). Conclusion Dahuang Zhechong capsules combined with entecavir have a better effect in the treatment of CHB patients with liver fibrosis compared with entecavir alone.

Objective To investigate the clinical efficacy and safety of Qinggan Huashi Huoxue decoction combined with liver- protecting and enzyme- lowering drugs in the treatment of alcoholic liver disease( ALD). Methods A total of 175 ALD patients who were admitted to The Second People's Hospital of Tangshan from January 2012 to December 2015 were enrolled and randomly divided into treatment group( 87patients) and control group( 88 patients). The patients in the control group were asked to quit smoking and were given nutritional support and medications including polyene phosphatidylcholine,reduced glutathione,magnesium isoglycyrrhizinate,and ursodeoxycholic acid capsules,and those in the treatment group were given the self- made traditional Chinese medicine Qinggan Huashi Huoxue decoction in addition to the therapeutic regimen for the control group. During the three courses of treatment( 12 weeks),the patients' clinical symptoms and signs were observed,liver function [alanine aminotransferase( ALT),aspartate aminotransferase( AST),gamma- glutamyl transpeptidase( GGT),albumin( Alb),and total bilirubin( TBil) ]and blood lipids [total cholesterol( TC),triglyceride( TG),high- density lipoprotein( HDL),and low- density lipoprotein( LDL) ] were performed regularly,the results of routine blood tests and abdominal ultrasound findings were recorded regularly,and adverse events which occurred during treatment were recorded. The independent samples t- test was used for comparison of continuous data between groups,an analysis of variance with repeated measures was used to compare the differences at each time point between the two groups,and the chi- square test was used for comparison of categorical data between groups. Results After treatment,all patients achieved varying degrees of improvements in clinical symptoms and signs,which showed significant differences between the two groups at weeks 4,8,and 12 of treatment( t = 14. 390,10. 487,and 13. 547,all P < 0. 05). The liver function parameters in the two groups showed varying degrees of improvements at weeks 4,8,and 12 of treatment,and ALT,AST,and GGT showed significant differences between the two groups at weeks 4,8,and 12 of treatment( F = 21. 050,8. 108,and 12. 038,all P < 0. 01). In the control group,HDL showed a significant change after treatment( t = 3. 101,P < 0. 05),and in the treatment group,HDL and LDL showed significant changes after treatment( t = 6. 818 and 2. 532,both P < 0. 05). After treatment,HDL showed a significant difference between the two groups( t = 2. 784,P < 0. 05). The overall response rate also showed a significant difference between the control group and the treatment group( 53. 4% vs 82. 8%,χ2= 28. 74,P < 0. 001). No patient experienced significant adverse events during treatment. Conclusion Qinggan Huashi Huoxue decoction combined with liver- protecting and enzyme- lowering drugs can improve the clinical outcome of ALD patients and holds promise for clinical application.

Objective To develop a novel drug lymphocyte stimulation test( DLST) using cell counting kit- 8( CCK- 8) assay,and to investigate its application in the diagnosis of acute drug- induced liver injury( DILI). Methods The patients with acute DILI who were admitted to Beijing You An Hospital from January 2011 to December 2014 were screened,and the suspected drugs for liver injury were collected. The patients' peripheral blood mononuclear cells( PBMCs) were isolated,cultured in vitro,and then incubated with the suspected drugs. The CCK- 8 assay was used to measure the proliferation of lymphocytes. A positive control group was established for each experiment and was stimulated by phytohemagglutinin. Meanwhile,PBMCs from healthy subjects were enrolled as the negative control group,and were exposed to the same drugs and measured simultaneously. The results of DLST were introduced into the Roussel Uclaf Causality Assessment Method( RUCAM) scoring system to assess their effects on the sensitivity of the diagnosis of acute DILI. The chi- square test was used for comparison of categorical data between groups. Results Preliminary experiments were performed to determine the experimental conditions of DLST based on CCK- 8 assay. A total of 61 patients with acute DILI were enrolled. DLST based on CCK- 8 assay was completed in 52 patients who received a total of 75 drugs,and the results were identified to be reliable in 48 patients with 70 drugs. A total of 19 patients with22 drugs achieved positive results of DLST,with positive rates of patients and drugs being 39. 6% and 31. 4%,respectively,and a specificity of 93. 8%. Thirteen( 38. 2%) out of 34 traditional Chinese medicines and 9( 25%) out of 36 Western medicines achieved positive results of DLST,and the median stimulation index was 2. 29( 1. 81- 14. 20). After the results of DLST were introduced into the RUCAM scoring system,the proportion of drugs assessed as“highly probable”or “probable”were increased from 74. 7% to 92. 0%( χ2= 8. 112,P =0. 004). Conclusion CCK- 8 assay can be used to perform DLST,and the introduction of DLST results into the RUCAM scoring system can significantly improve the sensitivity of RUCAM in the diagnosis of DILI. Compared with the conventional DLST,DLST based on CCK-8 has the advantages of few requirements for laboratories,a simple and timesaving procedure,and low costs,and can be further used and verified in clinical practice.

Objective To establish the autophagy model of normal human liver cell line 7702 induced by hypoxia and starvation,and to lay a foundation for further studies on the influence of autophagy on liver function. Methods The 7702 cells were selected and incubated with95% air and 5% CO2 at a temperature of 37 ℃( normal control group). The Binder three- gas incubator was used,with a temperature of37 ℃,a CO2 concentration of 5%,and an O2 concentration of 0. 3% to provide a hypoxic environment,and the serum- free DMEM was used to induce starvation. These cells were divided into 6-,12-,18-,and 24- hour hypoxia- starvation groups. Western blot was used to measure the protein expression of Beclin 1,Atg5,and LC3 in the normal control group and experimental groups,RT- q PCR was used to measure the mRNA expression of Beclin 1 and Atg5 in each group,and after transfection of LC3 plasmid,immunofluorescence assay was used to observe autophagy in each group. An analysis of variance was used for comparison of continuous data between groups,and the least significant difference t- test was used for further comparison between any two groups; the chi- square test was used for comparison of categorical data between groups. Results The 6- hour hypoxia- starvation groups had higher protein expression of Beclin 1,Atg5,and LC3 than the normal control group or other treated groups. Compared with all the other groups,the 6- hour hypoxia- starvation group showed significantly increased mRNA expression of Beclin 1 and Atg5,as well as significantly greater increases in the mRNA expression of Beclin 1and Atg5( all P < 0. 05). The hypoxia- starvation groups had significantly lower numbers of autophagosomes than the normal control group,and the 6- hour hypoxia- starvation group had the highest number of autophagosomes( all P < 0. 05). Conclusion Hypoxia and starvation established by physical methods can successfully induce hepatocyte autophagy,which is the most remarkable at 6 hours of hypoxia and starvation.

Objective To investigate the characteristics of gene mutation in Chinese patients with hereditary hemochromatosis( HH).Methods A total of 9 patients with HH who visited Beijing Friendship Hospital,Capital Medical University from January 2013 to December2015 were enrolled. The genomic DNA was extracted,and PCR amplification and Sanger sequencing were performed for all the exons of four genotypes of HH,i. e.,HFE( type Ⅰ),HJV( type ⅡA),HAMP( type ⅡB),TFR2( type Ⅲ),and SLC40A1( type Ⅳ) to analyze gene mutations. A total of 50 healthy subjects were enrolled as control group to analyze the prevalence of identified gene mutations in a healthy population. Results Of all patients,2 had H63 D mutation of HFE gene in type Ⅰ HH,1 had E3 D mutation of HJV gene in typeⅡA HH,2 had I238 M mutation of TFR2 gene in type Ⅲ HH,and 1 had IVS 3 + 10 del GTT splice mutation of SLC40A1 gene in type ⅣHH. No patients had C282 Y mutation of HFE gene in type Ⅰ HH which was commonly seen in European and American populations. Five patients had no missense mutation or splice mutation. In addition,it was found in a family that a HH patient had E3 D mutation of HJV gene,H63 D mutation of HFE gene,and I238 M mutation of TFR2 gene,but the healthy brother and sister carrying two of these mutations did not had the phenotype of HH. Conclusion HH gene mutations vary significantly across patients of different races,and non- HFE- HH is dominant in the Chinese population. There may be HH genes which are different from known genes,and further investigation is needed.

Portal hypertension is a common clinical disease and brings a series of complications including the formation of gastrointestinal varicose veins,ascites,hepatic encephalopathy,and abdominal varicose veins. Most of these complications are related to the opening of collateral circulation after the increase in portal venous pressure. On one hand,collateral circulation helps to alleviate the high portal venous pressure,and on the other hand,it brings related complications to patients. This article reviews recent reports and studies on collateral circulation related to portal hypertension,in order to increase our knowledge of collateral circulation in portal hypertension and improve clinical diagnosis and treatment of such disease.

Hepatocellular carcinoma( HCC) is one of the most common malignant tumors in the world. How to improve cure rate and reduce recurrence and metastasis of HCC have always been hot topics in research. T- helper cell 17( Th17),an important subset of CD4+T-helper cells,plays a critical role in infection,autoimmune response,and tumor microenvironment. This article introduces the discovery of Th17 and related cytokines,summarizes its mechanisms of action and clinical significance in the development and progression of HCC,and provides some clues for new targets in the treatment of HCC and prognosis prediction.