探讨慢性活动性肝炎诊断中腺苷脱氨酶检测的意义

目的：探讨慢性活动性肝炎（Chronic active hepatitis，CAH）诊断中腺苷脱氨酶（Adenosine deaminase ADA）血清化学指标检测的意义。方法：选择2012年2月-2012年8月在我院确诊为CAH的患者50例设为实验组，选择同期在我院进行健康体检合格者50例设为对照组，均进行ADA检测，同时检测与肝功能密切相关的血清学指标谷丙酸氨基转移酶（ALT）、总胆红素（TBIL）和白蛋白（ALB）。结果：实验组ALB水平明显低于对照组，差异具有统计学意义（p〈0.05），实验组ADA、ALT和TBIL水平明显高于对照组，差异具有统计学意义（p〈0.05）；CAH患者血清ADA的阳性检出率为86.0%以上，与肝功能相关的血清学指标比较，差异具有统计学意义（p〈0.05）。结论：ADA可以作为CAH发生、发展的重要备选指标，有助于提高CAH的阳性检出率，对判断CAH的病情变化和预后可能有一定的参考价值。

Dynamic tracking of stem cells in an acute liver failure model

AIM： To investigate a dual labeling technique, which would enable real-time monitoring of transplanted em- bryonic stem cell （ESC） kinetics, as well as long-term tracking. METHODS： Liver damage was induced in C57/BL6 male mice （n = 40） by acetaminophen （APAP） 300 mg/kg administered intraperitoneally. Green fluores- cence protein （GFP） positive C57/BL6 mouse ESCs were stained with the near-infrared fluorescent lipophilic tracer 1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbo- cyanine iodide （DiR） immediately before transplantationinto the spleen. Each of the animals in the cell therapy group （n = 20） received 5 x 106 ESCs 4 h following treatment with APAP. The control group （n = 20） re- ceived the vehicle only. The distribution and dynamics of the cells were monitored in real-time with the IVIS lumina-2 at 30 rain post transplantation, then at 3, 12, 24, 48 and 72 h, and after one and 2 wk. Immunohisto- chemical examination of liver tissue was used to identify expression of GFP and albumin. Plasma alanine amino- transferase （ALT） was measured as an indication of liver damage.RESULTS： DiR-stained ESCs were easily tracked with the IVIS using the indocyanine green filter due to its high background passband with minimal background autofluorescence. The transplanted cells were confined inside the spleen at 30 min post-transplantation, gradu- ally moved into the splenic vein, and were detectable in parts of the liver at the 3 h time-point. Within 24 h of transplantation, homing of almost 90% of cells was confirmed in the liver. On day three, however, the DiR signal started to fade out, and ex vivo IVIS imaging of different organs allowed signal detection at time-points when the signal could not be detected by in vivo imag- ing, and confirmed that the highest photon emission was in the liver （P 〈 0.0001）. At 2 wk, the DiRsignal was no longer detectable in vivo; however, immuno- histochemistry analysis of constitutively-expressed GFP was used to provide an insight into the distribution of the cells. GFP ＋ve cells were detected in tissue sections resembling hepatocytes and were dispersed throughout the hepatic parenchyma, with the presence of a larger number of GFP ＋ve cells incorporated within the sinu- soidal endothelial lining. Very faint albumin expression was detected in the transplanted GFP ＋re cells at 72 h; however at 2 wk, few cells that were positive for GFP were also strongly positive for albumin. There was a significant improvement in serum levels of ALT, albumin and bilirubin in both groups at 2 wk when compared with the 72 h time-point. In the cell therapy group, serum ALT was significantly （P = 0.016） lower and al- bumin （P = 0.009） was significantly higher when com- pared with the control group at the 2 wk time-point;however there was no difference in mortality between the two groups. CONCLUSION： Dual labeling is an easy to use and cheap method for longitudinal monitoring of distribu- tion, survival and engraftment of transplanted cells, and could be used for cell therapy models.

Decreased mitochondrial deoxyribonucleic acid and increased oxidative damage in chronic hepatitis C

AIM： To determine whether alteration of the mito- chondria DNA （mtDNA） copy number and its oxidative damage index （mtDNA△CT） can be detected by analysis of peripheral blood cells in hepatitis C virus （HCV）- infected patients. METHODS： This study enrolled two groups of pa- tients aged 40-60 years： a control group and an HCV- infected group in Department of Gastroenterology and Hepatology in Changhua Christian Hospital. Patients with co-infection with hepatitis B virus or human im- munodeficiency virus, autoimmune disease, malignant neoplasia, pregnancy, thyroid disease, or alcohol con- sumption 〉 40 g/d were excluded. HCV-infected pa- tients who met the following criteria were included： （1） positive HCV antibodies for 〉 6 mo; （2） alanine aminotransferase （ALT） levels more than twice the upper lim- it of normal on at least two occasions during the past 6 mo; and （3） histological fibrosis stage higher than F1. The mtDNA copy number and oxidative damage index of HCV mtDNA （mtDNA△CT） were measured in periph- eral blood leukocytes. The association between mtDNA copy number and mtDNA△CT was further analyzed using clinical data. RESULTS： Forty-seven normal controls （male/female： 26/21, mean age 50.51 ± 6.15 years） and 132 HCV- infected patients （male/female： 76/61, mean age 51.65 ± 5.50 years） were included in the study. The geno- types of HCV-infected patients include type 1a （n = 3）, type 1b （n = 83）, type 2a （n = 32）, and type 2b （n = 14）. Liver fibrosis stages were distributed as follows： F1/F2/F3/F4 = 1/61/45/25 and activity scores were A0/ A1/A2/A3 = 7/45/55/25. There were no age or gender differences between the two groups. HCV-infected pa- tients had higher hepatitis activity （aspartate transami- nase levels 108.77 ± 60.73 vs 23.19 ± 5.47, P 〈 0.01; ALT levels 168.69 ± 93.12 vs 23.15 ± 9.45, P 〈 0.01） and lower platelet count （170.40±58.00 vs 251.24 ± 63.42, P 〈 0.01） than controls. The mtDNA copy num- ber was lower in HCV-infected patients than in controls （173.49 vs 247.93, P 〈 0.05）. The mtDNA△CT was higher in HCV-infected patients than in controls （2.92 vs 0.64, P 〈 0.05）. To clarify the clinical significance of these results in HCV-infected patients, their association with different clinical parameters among HCV-infected pa- tients was analyzed. A negative association was found between mtDNA copy number and elevated aspartate transaminase levels （r = -0.17, P 〈 0.05）. Changes in mtDNA copy number were not associated with HCV RNA levels, HCV genotypes, liver fibrosis severity, or inflammatory activity in the liver biopsy specimen. How- ever, a correlation was observed between mtDNA△CT and platelet count （r = -0.22, P 〈 0.01）, HCV RNA level （r = 0.36, P 〈 0.01）, and hepatitis activity （r = 0.20, P = 0.02）. However, no difference in the change in mtDNA△CT was observed between different fibrosis stages or HCV CONCLUSION： Oxidative stress and mtDNA dam- age are detectable in patient＇s peripheral leukocytes. Increased leukocyte mtDNA△CT correlates with higher HCV viremia, increased hepatitis activity, and lower platelet count.

Intrahepatic expression of genes related to metabotropic receptors in chronic hepatitis

AIM： To screen for genes related to metabotropic re- ceptors that might be involved in the development of chronic hepatitis. METHODS： Assessment of 20 genes associated with metabotropic receptors was performed in liver speci- mens obtained by punch biopsy from 12 patients with autoimmune and chronic hepatitis type B and C. For this purpose, a microarray with low integrity grade and with oligonucleotide DNA probes complementary to target transcripts was used. Evaluation of gene expression was performed in relation to transcript level, correlation between samples and grouping of clinical parameters used in chronic hepatitis assessment. Clini- cal markers of chronic hepatitis included alanine and aspartate aminotransferase, ~,-glutamyltranspeptidase, alkaline phosphatase and cholinesterase activity, levels of iron ions, total cholesterol, triglycerides, albumin, glucose, hemoglobin, platelets, histological analysis of inflammatory and necrotic status, fibrosis according to METAVIR score, steatosis, as well as anthropometric body mass index, waist/hip index, percentage of adi- pose tissue and liver size in ultrasound examination. Gender, age, concomitant diseases and drugs were also taken into account. Validation of oligonucleotide microarray gene expression results was done with the use of quantitative real-time polymerase chain reaction （qRT-PCR）. RESULTS： The highest （0.002 〈 P 〈 0.046） expres- sion among genes encoding main components of metabotropic receptor pathways, such as the a subunit of G-coupled protein, phosphoinositol-dependent pro- tein kinase or arrestin was comparable to that of an- giotensinogen synthesized in the liver. Carcinogenesis suppressor genes, such as chemokine ligand 4, tran- scription factor early growth response protein 1 and lysophosphatidic acid receptor, were characterized by the lowest expression （0.002 〈 P 〈 0.046）, while the factor potentially triggering hepatic cancer, transcrip- tion factor JUN-B, had a 20-fold higher expression. The correlation between expression of genes of protein kinases PDPK1, phosphoinositide 3-kinase and protein kinase A （Spearman＇s coefficient range： 0.762-0.769） confirmed a functional link between these enzymes. Gender （P = 0.0046） and inflammation severity, mea- sured by alanine aminotransferase activity （P = 0.035）, were characterized by diverse metabotropic receptor gene expression patterns. The Pearson＇s coefficient ranging from -0.35 to 0.99 from the results of qRT-PCR and microarray indicated that qRT-PCR had certainlimitations as a validation tool for oligonucleotide mi- croarray studies. CONCLUSION： A microarray-based analysis of hepa- tocyte metabotropic G-protein-related gene expression can reveal the molecular basis of chronic hepatitis.

Metformin prevents hormonal and metabolic disturbances and 1,2-dimethylhydrazine-induced colon carcinogenesis in non-diabetic rats

Effects of two doses of the anti-diabetic drug, metformin （MF）, on hormonal and metabolic levels of serum of non-diabetic male Wistar rats with 1,2-dimethylhydrazine （DMH）-induced colon tumor adenocarcinomas were studied. Carcinogenesis in the animals was also observed. Rats with DMH-induced colon adenocarcinomas had elevated levels of serum glucose, insulin, insulin- like growth factor-l, total cholesterol, triglycerides, catalase, malonic dialdehyde, glycated hemoglobin, aspartate aminotransferase, and alanine aminotransferase and decreased hemoglobin. Treatment with two doses of MF normalized maiority of these changes in DMH-treated rats, whereas the drug was ineffective in rats without DMH treatment. The only exception was the decreased triglyceride levels in MF-treated rats. A 100 mg/kg dose of MF increased DMH-induced exophytic colon carcinomas and decreased endophytic tumors compared with untreated rats. Moreover, both MF doses increased DMH-induced and highly differentiated tumors and decreased the invasiveness of colon carcinomas compared with rats provided with DMH and water. Therefore, effects of MF on metabolic homeostasis are critical for preventing colon cancer.

Protective effects of C-phycocyanin on alcohol-induced acute liver injury in mice

Excessive alcohol consumption leads to liver disease. Extensive evidence suggests that C-phycocyanin(C-PC), a chromophore phycocyanobilin derived from Spirulina platensis, exerts protective eff ects against chemical-induced organ damage. In this study, we investigated whether C-PC could protect against ethanol-induced acute liver injury. Serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), triglyceride(TG), total cholesterol(CHOL), low-density lipoprotein(LDL), liver homogenate malondialdehyde(MDA), superoxide dismutase(SOD) content were measured, and pathological examination of liver sections were examined. C-PC showed obvious inhibitory eff ects on serum ALT, AST, TG, CHOL, LDL and MDA, and SOD content significantly increased in the liver. The structure of hepatic lobules was clear, liver sinus returned to normal, and liver cell cords were arranged in neat rows. Cloudiness, swelling, inflammatory cell infiltration and spotty necrosis of liver cells were significantly reduced. Therefore, C-PC can significantly protect against ethanol-induced acute liver injury.

Seasonal Variations in Serological Profiles and Growth Status of Farmed and Wild Clarias gariepinus （Burchell, 1822） Obtained from Asaba, Nigeria

Seasonal variations in serological profiles and growth status of farmed and wild Clarias gariepinus were investigated. Serological profiles of Clarias gariepinus can be an effective tool for proper monitoring of stress induced by management practices in intensive fish culture, which may elicit devastating effect on fish. Spectrophotometry procedures were used to analyze serum parameters of Clarias gariepinus. No different seasonal patterns were observed for mean values of albumin, total protein, urea, cholesterol, glucose and alanine amino-transferase. However, aspartate amino-transferase showed different seasonal pattern. Levels of cholesterol and alanine amino-transferase were highly significant in farmed and wild fish, separately. Juvenile fish had high level of urea irrespective of season. Seasonal variations in water quality parameters were observed except for pH which had no seasonal pattern. The growth exponential shows b-values between -0.048 and 7.434 for Clarias gariepinus. Adult female and juvenile fish from the wild had the highest b-value and the least b-value, separately. In this study, b-values were higher in the wild fish than the farmed fish. The condition factor for Clarias gariepinus ranged from 0.422 to 0.698, and was observed to be high in juvenile fish. With a condition factor less than 1, fish may not be doing well, probably due to environmental stress. Some serological parameters varied according to season and environment of fish. Thus, serological profile of fish is an effective and sensitive tool to monitor fish response to stress factors in the environment.

Prevention of Carbon Tetrachloride （CCl4）-Induced Liver Damage in Guinea Pigs by Cyphostemma digitatum

Cyphostemma digitatum （Vitaceae） is a perennial, climbing, succulent undershrub with compound fleshy leaves and tendrils. The plant is used mainly as a food flavoring, but it is also a main constituent of traditional Yemeni soup （Marak）. Besides that, it has been described to be used as a medicinal plant. The aim of this work was to study the hepatoprotective effect of the aqueous extract of C. digitatum against CCl4-induced liver injury in Guinea pigs. Animals were divided into four groups. Group I, served as normal control. Group II received 2 mL CCl4/kg b.w. diluted with olive oil, at 1：1 ratio on day 11. Group III （test group） was pre-treated orally with 100 mg/kg b.w. aqueous leaves extract of C. digitatum for 10 days followed by subcutaneous injection of CC14 （2 mL/kg b.w.）, once on day 11. Group IV were orally given Liv-52 （100 mg/kg b.w.） once daily for 10 days followed by subcutaneous injection of CC14. Our results show that, the activity of serum hepatic enzymes （alanine aminotranferase （ALT）, aspartate aminotranferase （AST）, and alkaline phosphatase （ALP）） were significantly elevated in Guinea pigs treated with CCl4, while both the C. digitatum extract and Liv-52 reduced significantly these enzymes activity. Also, the levels of glucose, urea, cholesterol and triglycerides were decreased when compared with intoxicated control. Histopathological examination of intoxicated animals showed fatty changes, inflammation and necrosis indicating liver damage, while the animals received C. digitatum or Liv-52 showed less pathological effects or normal liver when compared to animals treated with CC14 alone. Biochemical and histological results confirm the hepatoprotective effect of aqueous extract of C. digitatum.

Panax notoginseng saponins ameliorates experimental hepatic fibrosis and hepatic stellate cell proliferation by inhibiting the Jak2/Stat3 pathways

OBJECTIVE： To investigate the inhibitory effect of Panax notoginseng saponins（PNS） on liver fibrosis and explore the underlying mechanisms.METHODS： Carbon tetrachloride（CCl4）-treated rats and hepatic stellate cells（HSCs） were used. The effect of PNS on CCl4-induced liver fibrosis was studied with histochemical and biochemical analysis.Transforming growth factor（TGF）-β1, α-smooth muscle actin（α-SMA）, and collagen Ι m RNA expression were determined by reverse transcriptase-polymerase chain reaction（RT-PCR）. Mean-while, the protein expression levels of α-SMA, collagen Ι, phosphorylation-Janus activated kinase signal transducer（p-Jak2） / Jak2, and phosphorylation-activator of transcription（p-Stat）3 / Stat3 were determined by immunohistochemistry and / or immunoblotting.RESULTS： PNS treatment significantly improved the liver function of rats as indicated by decreased serum enzymatic activities of alanine aminotransferase and aspartate aminotransferase. Histopathological results indicated that PNS alleviated liver damage and reduced the formation of fibrous septa. Moreover, PNS significantly decreased liver hydroxyproline and significantly attenuated expressions of collagen p-Stat3 / StatⅠ, α-SMA, TGF-β1, p-Jak2 / Jak2,and 3 in the rat liver fibrosis model and HSCs.CONCLUSION： PNS can relieve liver fibrosis by modulating Jak2/Stat3 signaling transduction pathway, which may be one of its mechanisms to suppress hepatic fibrosis.

Clinical study on treatment of chronic viral cholestatic hepatitis with Chishaodanpi decoction

OBJECTIVE: To observe the therapeutic effect of Chishaodanpi decoction(CSDPD) on chronic viral cholestatic hepatitis.METHODS: A total of 107 subjects with chronic viral cholestatic hepatitis were enrolled in our hospital from March 2007 to November 2012. Patients were randomly divided into treatment(54 cases)and control groups(53 cases). The control group was treated with potassium magnesium aspartate,diammonium glycyrrhizinate, glucurolactone, vitamin C, and lamivudine, once a day. The treatment group was treated with modified CSDPD, 100 m L a time, twice a day, in addition to the treatment given to the control group. The patients in both groups were treated for 8 weeks. The main symptoms and signs were recorded every day throughout the clinical trial. Before and after the trial,changes in liver function including total bilirubin(TBil), direct bilirubin(DBil), total bile acid(TBA),and the activities of alkaline phosphatase(ALP), alanine aminotransferase(ALT), aspartate aminotransferase(AST), and γ-glutamyl transferase(γ-GT),were all detected. Adverse reactions were also recorded.RESULTS: There were no differences in gender, age,disease duration, symptoms, signs, or laboratory findings between the two groups(P>0.05). After an8-week treatment, improvements in jaundice, weakness, poor appetite, abdominal distention, and skin itching were significantly better in the treatment group than in the control group(P<0.05). In the treatment group, 43 patients had a significant response to the treatment, seven patients had a response, and four patients had no response, with 21,12, and 20 patients in the control group, respectively. The total effective rate was 92.6% in the treatment group and 62.3% in the control group, which was a significant difference(P<0.05). The levels of TBil, DBil, TBA, ALP, ALT, AST, and γ-GT in both groups were significantly lower after treatment,and were significantly different between the two groups(P<0.05). A few patients in the treatment group had mild adverse effects such as increased bowel movement frequency and mild stomachache. No other adverse reactions were observed in either group.CONCLUSION: CSDPD has a satisfactory therapeutic effect on chronic viral cholestatic hepatitis.

The 96-week clinical outcomes after cessation of nucleos(t)ide analog treatment in chronic hepatitis B patients

Background:Chronic hepatitis B(CHB)patients have a high virological relapse rate after cessation of nucleos(t)ide analog(NA)treatment,but the clinical outcome remains unclear.This study aimed to investigate the 96-week clinical outcomes and the risk factors for relapse in CHB after cessation of NAs.Methods:This study was a prospective trial;74 eligible patients were enrolled.The patients underwent NA cessation and follow-up according to the 2012 Asian Pacific Association for the Study of the Liver Guideline.Symptoms,biochemical(aspartate aminotransferase[AST],alanine aminotransferase[ALT],total bilirubin,urea nitrogen,creatinine),virological data(hepatitis B surface antigen[HBsAg],hepatitis B e antigen[HBeAg],hepatitis B e antibody[HBeAb],hepatitis B virus[HBV]DNA levels),and color Doppler ultrasound examination results were recorded and analysed.Results:After NA cessation,19 cases were HBsAg-negative without relapse during the 96-week follow-up.Of the 55 cases of HBsAg-positive after cessation,four types of clinical outcomes were observed.Twelve patients had no relapse during the 96-week follow-up(type A,21.8%),7 patients underwent virological relapses but spontaneously had a non-virological relapse(type B,12.7%),10 patients maintained virological relapse(type C,18.2%),and 26 patients turned to clinical relapse,received NA retreatment,and achieved ALT normalization and negative conversion of HBV DNA within 12 months(type D,47.3%).The 2-year overall cumulative rates of virological and clinical relapses were 58.1%and 24.3%,respectively.Independent factors associated with virological relapse were duration of negative HBV DNA,EOT(end of treatment)HBsAg,and original status of HBeAg.The EOT HBsAg was also an independent factor for clinical relapse.Conclusions:There are four types of clinical outcomes in patients with CHB after cessation of NA treatment.Further research is needed to explore the mechanism of different clinical outcomes.The EOT HBsAg level is an independent factor associated with both virological and clinical relapse.

Effect of Yajieshaba,a preparation of Dai indigenous medicine,on enhanced liver detoxification

OBJECTIVE:To explore the mechanistic effects of Yajieshaba(YJSB) on enhanced liver detoxification.METHODS:The effects of YJSB on alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were assayed in five acute chemical liver injury models[carbon tetrachloride(CCU),D-galactosamine(D-Glan),4-acetamidophenol(AAP),thioacetamide(TAA) and 1-naphthyl isothiocyanate(ANIT)].Sleep latency and sleep time of pentobarbital sodium were tested in control mice and CCL model miceafter oral YJSB administration.The effects of YJSB on drug metabolism enzymes of liver microsomes were tested in control rats and CCI_4model rats.The levels of cytochrome P450(CYP450) and Cyt b5 in liver microsomes were assayed using the method by Omura and Sato,and activities of erythromycin N-demethylase(ERD)and aminopyrine N-demethyl(ADM) were evaluated by Nash colorimetry.Probe substrate-based high performance liquid chromatography(HPLC)methods were established for CYP3A4 and CYP1A2.RESULTS:The level of serum ALT was reduced by YJSB at 3.51 g/kg in the five models as follows:CCl_4 > D-Glan,AAP,ANIT > TAA.YJSB treatment did not reduce the level of serum AST.YJSB at 3.51 g/kg prolonged the sleep latency in control mice and shortened the sleep time of control mice and CCl_4 model mice.For control rats,YJSB at 2.43 g/kg increased the levels of CYP450 and Cyt b5 and induced the activities of ERD and ADM;for liver injuries induced by CCI_4 in rats,YJSB at 2.43 g/kg increasedthe levels of CYP450 and Cyt b5.These results suggest that YJSB at 2.43 g/kg induces CYP3A4 and CYP1A2.CONCLUSION:These results suggest that YJSB enhanced liver detoxification and the mechanisms may be partially related to CYP3A4 and CYP1A2 induction.

A nano-functionalized real-time electrochemiluminescent biosensor for alanine transaminase assay

An electrochemiluminescent (ECL) biosensor was constructed for selective assay of alanine aminotransferase (ALT) based on the enzymatically catalyzed oxidation of pyruvate by pyruvate oxidase (PYOD). The composite of potassium ferricyanide and carbon nanotube was adopted to pre-functionalize the basal platinum electrode while the potassium ferricyanide acted as the activator of PYOD. The ALT catalyzed the reaction of L-alanine and-ketoglutarate to produce pyruvate which could be further enzymatically oxidized by PYOD to yield H2O2 to intensify the ECL of luminol. The biosensor showed rapid response for real-time measurement of ALT in the linear concentration range from 0.00475 to 350 U/L (r = 0.993) with a relatively standard deviation of 2.5% (CALT = 47.5 U/L,n = 6). The biosensor was applied to assay the ALT in rat serum with average recovery of 90.5%.

Effects of lifestyle modification on liver enzyme and Fibroscan in Indian patients with non-alcoholic fatty liver disease

Background and aims:Non-alcoholic fatty liver disease(NAFLD)is remarkably increasing in developing countries like India,in parallel with the increasing incidence of obesity.Lifestyle modification is a recommended treatment for NAFLD.In most of the previous studies,improvement after lifestyle modification was assessed by liver fibrosis through liver biopsy,but we cannot do a serial liver biopsy in every NAFLD patient.Liver fibrosis can also be assessed by fibroscan non-invasively in NAFLD.This study was designed to evaluate the effect of lifestyle modification on liver enzymes and Fibroscan values in a population with NAFLD.Methods:Initially,50 NAFLD patients were included in this prospective follow-up study;however,after 6 months of lifestyle modification,only 39 participants were studied.During both the first and second consultations,Fibroscan was carried out.All participants underwent a careful interview,anthropometry measurements and radiological and biochemical tests during every consultation.Results:After 6 months of lifestyle modification,Fibroscan values improved significantly(8.3160.11kPa vs 7.8760.12kPa,p¼0.009).Alanine aminotransferase(ALT)values also showed improvement during the second consultation(97.2562.62 U/L vs 66.6963.95 U/L,p<0.001).Conclusion:Measured by Fibroscan and liver enzymes,it has been found that lifestyle modification is an effective therapy to downgrade hepatic injury in NAFLD patients.Serial Fibroscan can be used to assess the treatment response in NAFLD patients due to its non-invasive nature.