Objective To observe the blood-lipid changes in patients with nonalcoholic fatty liver disease （NAFLD）. Methods This was a retrospective study of 638 patients in our hospital. We collected data including of liver function,blood-lipids,fasting blood-glucose（FBG）,HOMA-IR and hepatic CT scan. Results The blood-lipids in patients with NAFLD showed high level of TG. The degree of fatty liver was positively correlated with BMI and course of disease（P<0.05）. The levels of FBG,HOMA-IR,TC,APO-B,NON-HDL-C were increased gradually with the degree of fatty liver getting severe （P<0.05）. The levels of HDL-C,LDL,APO-A1 and TG had no obvious difference among the degree of fatty liver（P>0.05）. The level of ALT was positively correlated with degree of fatty liver,BMI and HOMA-IR,was negatively correlated with age（P<0.05）,but there was no difference in FBG and other blood-lipids. Conclusions Blood-lipids in patients with NAFLD showed high level of TG. The level of TC,APO-B and NON-HDL-C were increased gradually with the degree of fatty liver getting obvious,and there was no connection between ALT and blood-lipids.

Objective The aims of this study was to investigate the prevalence and risk factors of non-alcoholic fatty liver disease（NAFLD）in checked-up subjects in Guangdong Province. Methods 20047 subjects were checked-up by ultrasound and blood parameters. Results The prevalence of NAFLD in this subjects was 13.2%,in which the prevalence of NAFLD in male subjects was significantly higher than that in the female subjects（18.8% vs 7.8%,P<0.01）;the peak prevalence（23.8% to 25.6%） occurred in persons of 50- to 69-year old age,which were obviously higher than those in the other groups（4.7% in younger than 30,10.8% in younger than 40,17.6% in younger than 50 and 18.7% in older than 79 years of age,P<0.01）;Individuals with hypertension,hyperlipidemia and hyperglycemia had higher prevalence of NAFLD than those without（31.3% vs. 10.0%,41.3% vs. 8.2%,and 27.9% vs. 10.6%,P<0.01）. Conclusions The prevalence of NAFLD is high in Guangdong Province and there is a closed correlation of NAFLD to hypertension,hyperlipidemia and hyperglycemia.

Objective To investigate serum cytokeratin （CK18） levels in the diagnosis of patients with nonalcoholic fatty liver diseases（NAFLD）. Methods Ninety-three patients with NAFLD and forty-four healthy persons matched with age and gender were included in this study. Serum ALT,AST,GGT,CK18,FBG,FINS,TC,TG,HDL,LDL,DBP,SBP,height,weight,WC,HC,and waist hip ratio,body mass index and homoeostasis model assessment of insulin resistance were calculated. There were thirty patients with NAFLD undergoing liver bioposy. Results There was an significantly high serum CK18 levels in patients with NAFLD as compared with control subjects（P<0.001）;serum CK18 level in patients with elevated ALT group （16.38±8.1ng/ml） was higher than in normal ALT group（5.655±4.72ng/ml,P<0.001）;serum CK18 level（20.93±8.07ng/ml） in NASH group was higher than in NAFL group（7.61±5.51ng/ml,P<0.001）;Serum CK18 level was positively correlated to the degree of steatosis（r=0.527,P=0.003）,lobular inflammation（r=0.662,P<0.001）,and fibrosis（r=0.715,P<0.001）. Conclusion Serum CK18 level reflects the degree of lobular inflammation and fibrosis in NAFLD and has certain diagnostic value for NAFLD.

Objective To explore the effect of carnosic acid（CA）on high-fat diet（HFD）-induced obese non-alcoholic steatohepatitis（NASH）in rats. Methods Twenty-eight male Sprague-Dawley rats were randomly divided into control group （n=8） and experimental group（n=20）. After 10 weeks,rats fed with HFD were assigned to model group（n=10） and treatment group （n=10）,which were both fed with HFD and treated with intragastric administration of either NS or CA（200mg.kg^-1.d^-1） for 6 weeks. Results The rats in model group developed abdominal obesity and NASH in liver histopathology;the body weights in control and model groups were 449.5±47.4 g vs 531.8±40.4 g,visceral fat weight：4.51±1.63g vs 6.37±1.66g,TC：1.58±0.14 mmol/L vs 3.21±0.71mmol/L,ALT：36±18U/L vs 144±80U/L,AST：132±29U/L vs 218±64U/L,MDA:5.93±1.14nmol/ml vs 18.89±7.04nmol/ml,HOMA-IR:0.30±0.15 vs 0.65±0.34 and SOD:24.61±0.43U/ml vs 22.91±0.93U/ml（P<0.01）;Compared with those in the model group, improvement of all above indexes was found in CA-treated group. Conclusion Carnosic acid has satisfactory therapeutic effects on obesity and steatohepatitis induced with high-fat diet. Inhibiting the accumulation of visceral fat,regulating lipid metabolism,improving insulin resistance,resisting oxidative stress and lipid peroxidation damage may be part of its therapeutic mechanisms.

Objective To observe the influence of oral administration of ciprofloxacin on hepatic TLR4 signaling activated by metabolic endotoxemia in rats with nonalcoholic steatohepatitis （NASH） induced by high-fat diet. Methods Thirty male SD rats were divided randomly into model and intervention group fed with high-fat diet and normal group fed with normal diet. Rats in intervention group were administrated with oral ciprofloxacin from 9th week. At the end of 12th week,the endotoxin level in portal vein,fasting blood glucose and serum lipid were detected. NAS score was evaluated. The protein and mRNA levels of hepatic TLR4 and IRS-1 were detected by real time PCR or Western bloting. Liver and serum pro-inflammatory cytokines levels were tested by ELISA. Results Serum endotoxin level in model group was significantly higher than that in normal group（0.361±0.018 EU/ml νs. 0.324±0.013 EU/ml,P<0.01）;NAS score in ciprofloxacin-intervened group declined（4.40 ±0.26 vs. 6.9±0.3 in model,P<0.05）;TLR4 mRNA levels and its protein expression in model groups were 8.7 times and 1.4 times higher than that in normal group,and declined by 51% and 14% in intervention group;Compared with the normal group,IRS-1 mRNA and its protein expressions in model group declined by 69% and 47%,and increased 2.3 times and 1.6 times in intervention group;Serum and hepatic TNF-ɑ and IL-6 levels in model rats increased significantly （P<0.05）,which decreased in intervention groups. Conclusion Metabolic endotoxemia activates TLR4 signaling in liver and might play a role in the onset of NASH. Oral ciprofloxacin administration could reduce gut-derived endotoxin.

Objective The aim of this study was to evaluate the acceptability of FibroScan in detection of cirrhosis and the distribution of liver fibrosis in alcohol consulting out-patients. Methods 98 consecutive patients were included and needle liver biopsy was proposed when liver stiffness exceeded 13 kPa. Results The measurement could not be performed correctly in 9 patients because of excess weight. Of the 53 patients with liver stiffness greater than 13 kPa,8 refused liver biopsy and three were primarily managed for other diseases. In the 33 patients in whom liver biopsy was performed,the presence of cirrhosis was confirmed in 21 cases and liver fibrosis in 12 cases. Conclusion FibroScan allows the diagnosis of cirrhosis with a high positive predictive value （97%） and assesses non-invasively liver fibrosis. We can formulate the following hypothesis which remains to be confirmed: below 8 kPa,there is probably no significant fibrosis;For patients between 8 and 13 kPa,the hepatic fibrosis is accumulating;and finally,beyond 13 kPa,the cirrhosis is established and a specific treatment should be implemented.

Objective To study the effect of IGF-1 on intestinal mucous injury in mice with alcohol-induced liver disease and to observe the impact of glutamine on expression of IGF-1. Methods Thirty c57 mice were randomly divided into control,model and intervention group（10 in each）. The control group were fed with Lieber Decarli liquid without ethanol,and mice in the model and intervention group were fed with 4% ethanol Lieber Decarli liquid. At the same time,glutamine was given to the mice in the intervention group. The serum endotoxin was examined after 12 weeks,and intestinal tissues were assessed with HE staining and the expression of IGF-1 in intestine was detected by immunohistochemical method. Results The serum endotoxin in the model group was much higher than that in control group（0.38±0.05 Eu/L vs. 0.13±0.02 Eu/L,P<0.05）,while it decreased in intervention group as compared with the model group;the histopathological score of colon injury in the model group was much higher than that in control group（10.3±1.3 vs 4.8±1.2,P<0.05）;the expression of IGF in the model group was much higher than that in control group（2.3±0.2 vs. 0.9±0.2,P<0.05）. Conclusion Alcoholic liver disease is always complicated with intestinal mucous injury. IGF-1 might play a role in alcoholic liver disease. Glutamine can protect the intestinal mucosa,and it may be associated with the expression of IGF-1.

Objective To study the pathological changes of hepatic sinusoids in patients with chronic hepatitis B. Methods The number,total area,total perimeter and average diameter of hepatic sinusoids were detected by stereology under transmission electron microscope. Results The total area,total perimeter and average diameter of hepatic sinusoids in patients with chronic hepatitis B of severe degree increased,the basal lamina formed,and collagen deposited in Disse's space;WP bodies were detected in 53.5% and 59.1% in SECs in patients with CHB of mild and moderate degree,respectively,while that was only in 12.9% of patients with CHB of severe degree. Conclusion The pathological changes of hepatic sinusoids play a key role in the microcirculation disturbance of liver in patients with chronic hepatitis B.

Objective To measure the cognitive functions in patients with chronic hepatitis B （CHB）. Method Number connection test,digit symbol test,Stroop color-word test and clock drawing test were conducted in 196 patients with CHB and in 196 healthy controls. Results The score of clock drawing test in patients with CHB（22.8±4.9）was lower than that in healthy controls（23.9±4.0,t=-2.44,P<0.05）;While the results of other tests in the two groups were similar（P>0.05）. Conclusion CHB patients have executive dysfunction,but they do not show impairment in attention.

Objective To investigate the change of toll-like receptor 4 on peripheral blood mononuclear cells and the serum Thl/Th2 cytokine levels in patients with chronic hepatitis B receiving interferon-α therapy. Methods 50 patients with chronic hepatitis B were treated with interferon α for six months. After 12 weeks,they were devided into response and non-response group. Serum HBV DNA was detected by PCR;The Thl/Th2 cytokines were measured by ELISA;TLR4 was measured by flow cytometry. Results The IL-4 and TLR4 contents in the two groups at 3rd and 6th month were significantly lower than that before interferon-α therapy（P<0.05）,while serum IFN-γ levels were higher than that before interferon therapy（P<0.05）;The levels of IFN-γ,IL-4,TLR4,HBV DNA and ALT at 3rd month were significantly different between the two groups（P<0.05）;The levels of IFN-γ and IL-4 in 50 patients were not correlated with TLR4 level（P<0.05）;The change of TLR4 level in response patients at 3rd month was not correlated with the change of HBV DNA levels（P>0.05）. Conclusions The changes of IFN-γ and IL-4 level in CHB patients received 12 week interferon-α therapy may be not releted to the changes of TLR4 level;the changes of TLR4 level probably is releted to the early response in CHB patients received interferon-α therapy.

Objective To observe the 3 month-efficacy of adefovir dipivoxil in the treatment of patients with lamivudine-related hepatitis B viral YIDD or YVDD mutant infection. Methods 30 lamivudine-resistant chronic hepatitis B patients were treated by combination of adefovir dipivoxil and lamivudine in this study for three months. The serum HBV DNA was detected by fluorescent quantitation PCR. Results The HBV YIDD or YVDD mutant infection was found in 21 patients,and the mixed mutant and wild viral infection was found in 9;the baseline of serum HBV DNA,ALT and AST in mutant group were 6.2±1.4 lgcopies/ml,139.8±153.2U/L and 119.0±32.1U/L,respectively. After 3 month treatment,they dropped to 2.3±2.3lgcopies/ml,54.5±44.7U/L and 39.8±32.1U/L,respectively;the baseline of them in mixed infection group were 6.0±1.9 lgcopies/ml,144.7±128.2U/L and 114.2±131.1U/L,respectively and after 3 month treatment,they dropped to 2.4±2.4lgcopies/ml,55.8±53.2U/L and 42.8±32.3U/L,respectively（P>0.05）;the HBV DNA loss in mutant infection was found in 90.5%,serum ALT normalization in 90.5%,and the HBV DNA loss and ALT normalization in mixed infection were 88.9% and 77.8%,respectively;no HBeAg seroconversion was found in mutant infection group,while 37.5%（3/8）had HBeAg seroconversion in mixed infection group. Conclusion Conbination of lamivudine and adefovir dipivoxil therapy for patients with hepatitis B mutant or mutants and wild viral infection can get the same short-term efficacy.

Objective To determine the distribution of hepatitis C virus genotypes in patients with chronic hepatitis C,and to study the serum HCV RNA levels in patients with different HCV genotype infection. Methods The levels of serum HCV RNA was measured with real-time fluorescent quantitative reverse-transcription polymerase chain reaction,the serum anti-HCV was detected by ELISA,ALT by automatic biochemistry analyzer,the levels of hyaluronic acid,laminin,procollagen type Ⅲ N-terminal peptide and collage type Ⅳ were detectd by chemiluminescence iummunoassay and the HCV genotypes in 218 patients with chronic hepatitis C were determined by gene chip. Results There were nine different subgenotypes of HCV in 218 samples,including genotype lb,2a,3a,3b and 6 genotype in 208 sera and lb+2a,lb+3b,lb+6,2a+3a in other 10 sera,respectively;Genotype 1b was the main one（77.1%,168/218）,and the genotype 2a was the second（8.7%,19/218）;There was no significant difference between the serum HCV RNA levels in 208 patients with different HCV genotype infection（F=0.932,P>0.05）;There was no significant difference between HCV genotype and gender as well as the levels of four blood markers of hepatic fibrosis in 168 patients with 1b and 40 with non-genotype 1b infection（x²=0.857,P>0.05）. Conclusions HCV 1b is the main prevalent genotype in patients with chronic hepatitis C,and the serum level of HCV RNA is not related to HCV genotype infection.

Objective To observe the clinical manifestation,laboratory examinations and therapy of patients with human parvovirus B19 infection. Methods 19 hospitalized patients in our hospital from August,2008 to July,2010 infected with human parvovirus B19 were investigated by clinical presentation,laboratory examinations and related therapy. Results The 19 hospitalized patients infected with human parvovirus B19 mainly presented symptoms with fatigue （12 cases）,jaundice （10 cases）,splenomegaly（10 cases）,fever （10 cases）,rash （6 cases）,myalgia and arthralgia（6 cases）. Six patients out of the 19 patients had following diseases or complications,such as pregnancy（1 case）,acute liver failure（2 cases）,schizophrenia（1 case）,acute myelosuppression（1 case） and pneumonia （1 case）. The characteristics of liver dysfunction showed elevated liver enzymes（AST/ALT）,mild or moderate jaundice,decreased PTA and normal CHE. The laboratory examinations in the 19 hospitalized patients with human parvovirus B19 infection were got back to normal and they all recovered after active symptomatic treatment （protecting liver function,decreasing liver enzymes,decreasing jaundice and administration of prednisolone and gamma globulin in some patients）. Conclusion Human parvovirus B19 is the only agent resulting in human diseases in Genus Parvovirus. The human parvovirus B19 can cause a series of clinical diseases including erythema infectiosum,natural abortion,myelosuppression and arthritis,etc. This article demonstrates that human parvovirus B19 is correlated with human hepatitis. Human parvovirus B19 might be an important pathogen for non hepatitis A to E. Human parvovirus B19 can results in hepatic lesion and cause acute hepatitis or acute fulminant hepatitis. It seemed that the hepatic lesion caused by human parvovirus B19 is an acute/subacute benign process and is prone to natural self-recovery.

Objective To explore the effect of TGF-β1 on cell proliferation,cell cycle regulation and collagen expression by HSC-T6 cell line. Methods The HSC-T6 cell proliferation was detected by MTT at 24 and 36 hours, respectively;The HSC- T6 cells were treated with TGF-β1（10 ng/ml） or not as control group. Flow cytometry was performed to measure the cell cycle;Fluorescent quantitative real time RT-PCR was used to quantify the mRNA levels of SMAD3,c-myc,cdk-2,cyclinE,EGF,HGF,Bcl-2,NF-κB,MMP1,MMP9,MMP14,TIMP-1,PAI-1,α-SMA,Collagen-I and Collagen-Ⅲ genes;Collagen-I,Collagen-Ⅲ,and α-SMA secreted by HSC-T6 cells were detected by ELISA. Results In comparison to HSC-T6 cells in control group,TGF-β1 decreased the cell counts in G₀-G₁ phase（24h: 57.3±8.5% vs 60.6±9.7%;36h: 53.0±2.2% vs 56.6±5.0%,both P>0.05）,while it increased the cells in S phase（24h:30.6±7.2% vs 26.4±10.1%;36h:35.2±3.7% vs 30.8±2.5%,both P>0.05）;We found that TGF-β1 treatment increased the mRNA levels of SMAD3,c-myc,cdk2,cyclin E, EGF,Bcl-2,NF-κB,TIMP1,PAI-1,α-SMA,and Collagn-I after treatment for 24 and 36 hours. Conversely,the mRNA level of HGF,MMP1,MMP9,MMP14,and collagen-III decreased at 24 hr,but increased at 36 hr. TGF-β1 promoted the secretion of collagen-I [24h:63.0±7.4ng/ml vs. 33.2±10.8ng/ml,P<0.05;36h:58.5±6.0ng/ml vs. 42.2±6.3ng/ml,P<0.05],and α-SMA[24h: 20.6±2.6ng/ml vs 4.2±0.7ng/ml,P<0.05;36h:59.7±14.6ng/ml vs 36.8±5.6ng/ml,P<0.05] by HSC-T6 cells after the treatment. Conclusion TGF-β1 can promote cell proliferation and the secretion of collagen of rat HSC-T6 cell lines.

Objective To establish a HBV infection model of CXCR3 knocked-out mice. Methods After CXCR3 knocked-out，nine of them and nine wild-type C57BL/6 mice were injected hydrodynamically ten micrograms of pAAV/HBV1.2 DNA into the tail veins. Then，the mice were regularly bled to monitor the serum levels of HBsAg，HbeAg and HBV DNA. The HBcAg in the livers from injected mice were detected by immunohistochemistry. Results All the bred CXCR3 knocked-out mice were homozygous. The serum levels of HBsAg from the CXCR3 knocked-out mice and the wild type C57BL/6 were 1134.69±244.42 and 1759.63±881.20（P=0.096） at the first day after the injection of the pAAVHBV1.2 plasmids；5305.29±1395.06 and 7493.29±658.63（P=0.003） at the fourth day，1615.04±1187.16 andm1536.19±1046.02（P=0.905） at the fifteenth day，and 229.45±79.27 and 228.19±295.02（P=0.996） at fortieth day，respectively；the serum levels of HBeAg from them were 6.65±1.50 and 20.61±4.03（P=0.000） at the first day，6.33±1.61 and 9.79±2.31（P=0.007） at the fourth day，3.52±1.97 and 2.85±0.74（P=0.425） at the fifteenth day，and 1.28±0.06 and 1.90±1.01（P=0.431）at fortieth day，respectively and the serum levels of HBV DNA from them were 4.38±0.22 lgcopies/ml and 6.56±0.16 lgcopies/ml（P=0.008） at day10，4.41±0.88 lg copies/ml and 5.69±0.04 lg copies/ml（P=0.177） at day15，4.48±0.04 lgcopies/ml and 6.44±0.16nlgcopies/m（P=0.004） at day25，and 3.66±0.45 lgcopies/ml and 5.20±0.28 lg copies/ml （P=0.055） at day40，respectively;HbsAg，HbeAg and HBV DNA in CXCR3 knocked-out mice were continuously positive untill the day 40 after the injection of the pAAV/HBV1.2 DNA;Both cytoplasmic and nuclear HBcAg were detected in the livers of the CXCR3 knocked-out mice at the day 4，15，40 after the infection. The positive serum HBsAg，HbeAg and HBV DNA in CXCR3 knocked-out mice were similar to those in the wild type C57BL/6 mice. Conclusion We had successfully established a HBV infection model in CXCR3 knocked-out mouse， which might be helpful way to investigate the relationship between the CXCR3 and its ligands to the HBV infection.

Objective To investigate the hepatic expression of KCNJ15 in mice with MHV-3-induced fulminant hepatitis. Method MHV-3-induced fulminant hepatitis model was established and the hepatic expression of KCNJ15 were assayed by quantitative PCR and immunohistochemistry. KCNJ15 expressions on subgroups of intrahepatic lymphocytes were determined by flow cytometry. Results The enhanced expression of KCNJ15 in liver tissue was observed,especially at 72h after MHV-3 injection. Moreover,the percentage of KCNJ15-producing CD4⁺T cells in the liver at 48h post infection increased to 25.17±7.68%,which was remarkably higher than that of at 0h（3.92±1.33%,P<0.001）;Similarly,hepatic KCNJ15-producing CD8⁺T cells was also increased to 37.08±8.73% with a significant difference as comparing to that of at 0h（6.98±3.48%,P<0.001）;Meanwhile,hepatic NK cells expressed KCNJ15 increased significantly from 7.72±1.34% at 0h to 19.80±4.25% at 24h post infection（P<0.001）. Conclusion The increased expression of KCNJ15 in liver may correlate with immune regulatory function of hepatic lymphocytes,which contributes to virus induced liver failure.

Objective The arm of this study was to investigate the changes of TCRγδ+CD3+CD4-CD8 （TCRγδ+DNT） cells in C3H/Hej mice with murine hepatitis virus type 3（MHV-3）- induced viral hepatitis. Methods MHV-3- induced viral hepatitis was established. At day 0,3,7,10,15 and 20 after MHV-3 infection,the TCRγδ+DNT cells in peripheral blood and spleen were counted by fluorescent antibodies staining and detected by flow cytometry for its proportion to CD3+T cells and the expression of CD25,CD28,CD30 and CD44 of the cells were identified. Results After MHV-3 infection,the proportion of TCRγδ+DNT to CD3+T cells in peripheral blood and spleen were markedly increased（P< 0.05） and it peaked at day 10（4.8± 1.5% and 4.5± 1.3% in peripheral blood and spleen,respectively）;The analysis of cell surface marker demonstrated that both of the phenotypes of TCRγδ+DNTs in peripheral blood and spleen were TCRγδ+CD3+CD4-CD8-CD25-CD28-CD30-CD44+. Conclusions The increase of TCRγδ+DNT in peripheral blood and spleen may play an important role in the pathological process of MHV-3-induced viral hepatitis.

Objective To investigate the inhibition of apoptosis of HepG-2 cells by bacalin in vitro. MethodsHuman hepatocellular carcinoma cells,HepG-2 were cultured with different concentrations of baicalin and the inhibitory rate of cell proliferation was calculated with MTT assay;Morphology changes were detected by Hoechst33258/PI double fluorescence coloration;The apoptosis rate was detected by TUNEL;the Caspase-9,Caspase-3 and Bcl-2 protein expression were detected by Western blot. Results The cell proliferation was inhibited by baicalin at concentration of 25 to 100μg/ml;the inhibitory rate in 48h was 50.63% at 50μg/ml and 77.62% at 75μg/ml,and the inhibitory effect was in the concentration and time-dependent manner;there was a typical apoptosis at 50μg/ml in 48h,showing cell shrinkage and fragmentation of cell nucleus;the caspase-9 and caspase-3 expression increased and Bcl-2 decreased as the concentration of baicalin increased. Conclusion Baicalin can inhibit proliferation of HepG2 cells by inducing apoptosis and the mechanisms might be related to the mitochondrial pathway.