AIM: To investigate the influence of compatibitity in Fengshi Maqian Tablet [ABD group(Semen strychni,Bombyx batryticatus,Herba ephedrae,Scorpio,Rhizoma atractylodis,Radix et Rhizoma glycyrrhizae) E group(Olibanum;Myrrha)]. METHODS: In the use of orthogonal design(5 factors and 2 levels).HPLC method were used to determine the main chemical componets such as strychnine,brucine and ephedrine.And anti-inflammation was adopted as pharmacological experiment. RESULTS: In the inhibitive experiment of granuloma,ABD group was superb to E group.In the tumescence experiment,otherwise. CONCLUSION: The minister,assistant and guide herbs in Fengshi Maqian Prescription had a significant effect on the toxicity of monarch medicines-Semen strychni.

Background: Early diagnosis and staging of liver fibrosis are important for the prognosis and evaluating the survival of patients.Aims: To systematically assess the diagnostic value of transient elastography (TE) for staging of liver fibrosis in patients with chronic liver disease.Methods: PubMed,Embase,Cochrane Library,CNKI,Wanfang and VIP from Jan.2001 to Dec.2015 were retrieved to collect the articles with staging of liver fibrosis in patients with chronic liver disease by TE.Data extraction was conducted.Article quality was evaluated by quality assessment of diagnostic accuracy studies 2 (QUADAS2).Meta-analysis was conducted by Stata 12.0 software.Results: Twenty articles involving 5 748 patients were included.Meta-analysis showed that the combined sensitivity,specificity and AUC of TE for diagnosing significant fibrosis (≥F2) were 0.78 (95％ CI: 0.73-0.82),0.85 (95％ CI: 0.80-0.88) and 0.88 (95％ CI: 0.85-0.91),respectively.The combined sensitivity,specificity and AUC for advanced fibrosis (≥F3) were 0.89 (95％ CI: 0.86-0.91),0.88 (95％ CI: 0.85-0.91) and 0.94 (95％ CI: 0.92-0.96),respectively.The combined sensitivity,specificity and AUC for cirrhosis (F4) were 0.91 (95％ CI: 0.86-0.95),0.89 (95％ CI: 0.87-0.92) and 0.95 (95％ CI: 0.93-0.97),respectively.Conclusions: TE technique has a good diagnostic value in assessing significant fibrosis,advanced fibrosis and cirrhosis in patients with chronic liver disease,especially for advanced fibrosis and cirrhosis.

OBJECTIVETo investigate whether gene expression profiles can be used to determine risk genes and predict HBV-related cirrhosis progression to liver carcinoma using Significance Analysis of Microarray (SAM) and Prediction Analysis of Microarray (PAM) methods.

METHODSThe Affymetrix GeneChip was used to establish the gene expression profiles of liver tissues from 15 patients with chronic hepatitis B and cirrhosis or hepatocellular carcinoma (HCC). Differentially expressed genes (fold-change more than 2; P value less than 0.01) were selected by GeneSpring GX software. Risk genes related to cirrhosis and liver carcinoma were generated by SAM and PAM methods. Real-time PCR was used to verify the expression of risk genes in the liver tissues.

RESULTSSamples were clustered into the cirrhosis subgroup (n =15) or the HCC subgroup (n =15). A total of 497 differentially expressed genes were identified, SAM identified 162 significant genes, including 18 up-regulated genes and 144 down-regulated genes (fold-change:-1.46 to 1.28). PAM identified 22 genes with a "poor risk signature" (defined with a threshold of 5.5), which were associated with classifying cirrhosis and liver carcinoma; of these risk genes, 4 were down-regulated and 18 were up-regulated in the HCC group compared to the cirrhosis group (fold-change: 2.038 to 7.897, P value less than 0.01). The correction of classification was more than 80% . FOXP1, SPINK1 and KCNJ16 were verified by real-time PCR as differently expressed in the two subgroups (P value =0.011, 0.002 and 0.004, respectively).

CONCLUSIONThe altered gene profiles of carcinogenesis in HBV-related cirrhosis involves hundreds of genes. The combination of three "poor risk genes" may represent potential targets for diagnosis and prediction of liver carcinoma progression.

Adult ; Carcinoma, Hepatocellular ; genetics ; pathology ; Female ; Gene Expression Profiling ; Hepatitis B, Chronic ; complications ; genetics ; pathology ; Humans ; Liver Cirrhosis ; genetics ; pathology ; virology ; Liver Neoplasms ; genetics ; pathology ; Male ; Microarray Analysis ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Transcriptome

OBJECTIVETo investigate the role of TGF-beta1 and STAT3 signaling in liver fibrosis using a rat model system and to determine the therapeutic mechanism of AG490 in relation to this signaling pathway.

METHODSRats were randomly divided into a control group and DENA-induced liver fibrosis model group, and then subdivided into AG490 treatment groups. During fibrosis development, liver tissue samples were collected at different time points (0, 4 and 8 weeks) and evaluated according to the Scheuer scoring system. Expression of STAT3, TGFbeta1, alpha-SMA, E-cadherin, MMP2 and TIMP1 was measured by PCR (mRNA) and immunohistochemistry and western blotting (protein).

RESULTSIncreasing degrees of inflammation and fibrosis were observed in liver tissues of DENA-treated rats throughout model establishment. The mRNA expression of TGFbeta1 and STAT3 was significantly increased in DENA-induced rats with advanced fibrosis (AF) compared to those with early fibrosis (EF) (P = 0.034 and P = 0.012 respectively). The protein expression of TGF-beta1, phospho-Smad2, alpha-SMA, E-cadherin, STAT3 and phospho-STAT3 was significantly increased in DENA-induced rats with AF compared to the unmodeled control group (P = 0.048, P = 0.003, P = 0.002, P = 0.028, P = 0.009 and P = 0.039). The protein expression of E-cadherin was lower in the DENA-induced rats with AF than in those with EF (P = 0.026). STAT3 and TGF-beta1 co-expression was detected in AF tissues. DENA-induced AG490-treated rats with AF showed substantially lower protein expression of STAT3, TGF-beta1, MMP2 and TIMP1 compared to DENA-induced untreated rats with AF (P = 0.006, P = 0.018, P = 0.010 and P = 0.005); in addition, the degrees of fibrosis and inflammation were also greatly reduced in the DENA-induced AG490-treated rats with AF compared to DENA-induced untreated rats with AF (P = 0.042 and P = 0.021). Conclusions STAT3 signal transduction may regulate the TGF-beta1 pathway and affect liver fibrosis, especially in the advanced phase. AG490 can inhibit TGFbeta1-STAT3 signaling, resulting in reversal of liver fibrosis.

Animals ; Disease Models, Animal ; Liver Cirrhosis ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Transforming Growth Factor beta1 ; metabolism ; Tyrphostins ; pharmacology

OBJECTIVETo investigate the epidemiological status of cholestasis in first-hospitalized patients with chronic liver disease in Shanghai, and to provide a scientific basis for developing prevention and treatment measures.

METHODSFrom April 2005 to September 2014, 5,146 first-hospitalized patients in Shanghai with a diagnosis of chronic liver disease were enrolled in this study. Clinical data of the 4,660 patients who fit the study criteria for participation were collected for retrospective analysis.Diagnosis of cholestasis was made according to serum alkaline phosphatase (ALP) levels higher than 1.5 times the upper limit normal (ULN) and gamma-glutamyltransferase (GGT) levels higher than 3 times the ULN. The incidence rate of cholestasis was assessed for relation to age, sex, etiology, and type of liver disease, and statistically compared to the general clinical data and specific biochemical indicators with potential sex-related differences. T-test and chi-square test were performed for the statistical analyses.

RESULTSOf the 4,660 study participants, 10.26% had cholestasis; the prevalence of cholestasis increased with increasing age in male patients. The distribution of the cholestasis incidence according to the type of chronic liver disease was: 75.00%, primary sclerosing cholangitis; 42.86%, primary biliary cirrhosis; 35.97%, hepatic tumor; 30.77%, autoimmune hepatitis; 28.31%, drug-induced liver disease; 16.46%, alcoholic hepatitis; 13.98%, cryptogenic cirrhosis; 12.99%, schistosomal cirrhosis; 7.53%, alcoholic cirrhosis; 7.32%, mixed cirrhosis; 5.94%, viral liver cirrhosis; 2.70%, nonalcoholic fatty liver disease. There was no significant difference in the prevalence of cholestasis between the two sexes. In the patients with cholestasis, the levels of GGT and total bilirubin were significantly different between the two sexes.

CONCLUSIONThe incidence rate of cholestasis in first-hospitalized patients with chronic liver disease was 10.26%, and the rate increased with increased age. Patients with primary sclerosing cholangitis or primary biliary cirrhosis had higher incidence rates of cholestasis. Incidence rates of cholestasis of the various chronic liver diseases were not related to sex.

Bilirubin ; China ; Cholestasis ; Chronic Disease ; Humans ; Incidence ; Liver Diseases ; Male ; Prevalence ; Retrospective Studies ; gamma-Glutamyltransferase

Objective: To observe the effect of poloxamer 188 (P188) on megakaryocyte cultivation and induction from cord blood mononuclear cells in order to obtain more megakaryocyte progenitor cells (MPC). Methods: The cord blood mononuclear cells were isolated and inoculated in cell culture bag or cell culture flask respectively. The WIGGENS shaker and cell culture bags were used to mimick WAVE Bioreactor for three-dimensional (3D) cell culture, and the P188 was added to induction medium, The cells were detected for morphology, surface marker, viability, and number on day 14. Results: In the two-dimensional (2D) culture, CD41⁺, CD41⁺/CD61⁺, CD61⁺ megakaryocytic numbers increased significantly after adding P188 (all P<0.01). And in the 3D culture of adding P188, the cell volume became larger and the nuclear shape was irregular, the cytoplasm appeared magenta granules, and the megakaryocyte cells became more mature. By 3D culture, the expression of CD41/CD61 was (36.30±1.27)% vs (23.95±1.34)%, hence the differentiation for MPC was significantly higher than that in the 2D group (P<0.01). Furthermore, adding P188 in 3D culture resulted in highest differentiation efficiency for MPC [(59.45±1.20)%]. There were no significantly differences in terms of cell viability and cell number among 3D culture containing P188, 2D and 3D culture groups (all P>0.05). Conclusion 3D culture was beneficial for the differentiation of MPC, but the cell viability was lower than 2D group; However, the satisfied cell growth and better induction efficiency were obtained by adding of P188, which might provide a new method of megakaryocytes production for clinical application.

Objective: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. Methods: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivation model were chosen for granulocyte induction. Cell morphology was observed by microscopy, and cell phenotype was detected by flow cytometry. The CD18 expression of granulocytes was tested by immunofluorescence assay, and phagocytosis test was executed as well. Results: Compared to fetal bovine serum (FBS) treatment group, cell viability, counts and differentiation rate of granulocytes induced by X-VIVO^TM 15 combined with TPO, SCF, G-CSF but without FBS were superior. And X-VIVO^TM15 medium was better than SCGM medium at effectiveness and cost. Using two-stage mode of hematopoietic stem cell expansion followed by granulocyte induction with X-VIVO^TM15 combining TPO, SCF and G-CSF, cell proliferation was nearly 132 times at day 21. Flow cytometry showed that the differentiation was lagged in 2-stage mode than in direct induction mode, CD15 expression was (69.60± 1.06) % vs (97.73±0.39) %; Wright-Giemsa staining demonstrated mature granulocytes; immunofluorescence showed the expression of lysosomal proteins CD18. A strong phagocytic function of mature granulocytes was demonstrated by phagotrophic efficiency of (51.43±0.05) %. And granulocyte had chemotaxis ability under the role of chemotactic factor IL-8. Conclusion Optimized culture media and cultivation mode are achieved for functional granulocytes induction in vitro.