中西医结合治疗肝损型肾综合征出血热48例

肾综合征出血热(HFRS)病毒感染常引起肝损害,加重病情。1996年1月～1998年12月,我们以大丹清瘟汤为主采用中西医结合早期治疗肝损型肾综合征出血热48例,效果较好,报道如下。 1 资料和方法 1.1 病例选择 按1986年南京HFRS会议制定的诊断标准,选择发病5日之内、经测HFRS IgM(+)患者;并具备以下几点:①血清肝炎病毒指标均阴性:②ALT值>64U/L;③可伴有黄疸,TBil>17μmol/L;④可伴有肝肿大、肝触痛或叩痛。随机分为治疗组和对照组。治疗组48例,男35例。

荣筋通痹汤治疗腰椎间盘突出所致肝肾虚损型坐骨神经痛的临床观察

目的:基于随机对照研究,评价荣筋通痹汤对腰椎间盘突出所致坐骨神经痛患者(肝肾虚损型)的疗效及安全性。方法:选取2019年1月至2022年7月北京积水潭医院中医科收治的符合入组标准的患者90例,按随机数字表法分为两组,各45例。研究过程中脱落19例,最终观察组41例、对照组30例,观察组予荣筋通痹汤口服,对照组予洛索洛芬钠片口服,治疗28 d。观察治疗前后两组的视觉模拟评分(VAS)评分、日本骨科协会下腰痛评价量表评分(JOA)评分、日常生活能力量表(ADL)量表评估,中医临床症状、主观症状及临床体征评分的差异;并检测肝肾功能及胃肠道不良反应,进行安全性评估。结果:治疗前观察组与对照组的分值差异无统计学意义;治疗后观察组VAS及ADL评分差值绝对值高于对照组(P<0.05)。两组JOA评分差值绝对值差异无统计学意义,观察组中医临床症状评分差值绝对值高于对照组(P<0.01);两组主观症状及临床体征评分差值绝对值差异无统计学意义。试验前后两组肝肾功能指标水平均正常,胃肠道反应差异无统计学意义。结论:荣筋通痹汤对腰椎间盘突出引起的坐骨神经痛肝肾虚损型患者的症状有明显改善作用,治疗后患者在疼痛、活动等方面均改善明显,且安全性可靠,有利于身体功能恢复。

白帽伞毒蕈中毒的中西医结合护理体会

对１７例毒蕈中毒患者，采用中西医结合治疗和护理，获得了治愈率达７６．４７％的良好效果。认为高质量的护理与协调统一的医护关系是抢救危重病人不可缺少的重要环节。

Patatin-like phospholipase domain containing-3 gene I148M polymorphism, steatosis, and liver damage in hereditary hemochromatosis

AIM： To investigate whether the patatin-/ike phosph- olipase domain containing-3 gene （PNPLA3） I148M polymorphism is associated with steatosis, fibrosis stage, and cirrhosis in hereditary hemochromatosis （HH）. METHODS： We studied 174 consecutive unrelated homozygous for the C282Y HFE mutation of HH （C282Y＋/＋ HH） patients from Northern Italy, for whom the presence of cirrhosis could be determined based on histological or clinical criteria, without excessive alcohol intake （〈 30/20 g/d in males or females） or hepatitis B virus and hepatitis C virus viral hepatitis. Steatosis was evaluated in 123 patients by histology （n = 100） or ul- trasound （n = 23）. The PNPLA3 rs738409 single nucle- otide polymorphism, encoding for the p.148M protein variant, was genotyped by a Taqman assay （assay on demand, Applied Biosystems）. The association of the PNPLA3 I148M protein variant （p.I148M） with steatosis, fibrosis stage, and cirrhosis was evaluated by logistic regression analysis. RESULTS： PNPLA3 genotype was not associated with metabolic parameters, including body mass index （BMI）, the presence of diabetes, and lipid levels, but the pres- ence of the p.148M variant at risk was independently associated with steatosis [odds ratio （OR） 1.84 per p.148M allele, 95% confidence interval （CI）： 1.05-3.31; P = 0.037], independently of BMI and alanine amino- transaminase （ALT） levels. The p.148M variant was also associated with higher aspartate aminotransferase （P = 0.0014） and ALT levels （P = 0.017） at diagnosis, independently of BMI and the severity of iron overload. In patients with liver biopsy, the 148M variant was independently associated with the severity （stage） of fibrosis （estimated coefficient 0.56 ± 0.27, P = 0.041）. In the overall series of patients, the p.148M variant was associated with cirrhosis in lean （P = 0.049）, but not in overweight patients （P = not significant）. At logistic regression analysis, cirrhosis was associated with BMI 〉~ 25 （OR 1.82, 95% CI： 1.02-3.55）, ferritin 〉 1000 ng/mL at diagnosis （OR 19.3, 95% CI： 5.3-125）, and with the G allele in patients with BMI 〈 25 （OR 3.26, 95% CI： 1.3-10.3）. CONCLUSION： The PNPLA3 I148M polymorphism may represent a permissive factor for fibrosis progression in patients with C282Y＋/＋ HH.

Association between TRAIL expression on peripheral blood lymphocytes and liver damage in chronic hepatitis B

AIM:To explore a novel mechanism for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), upregulation of CD4+ and CD8+T lymphocytes participating in the patho-physiological process of chronic hepatitis B (CHB). METHODS: The levels of serum soluble TRAIL (sTRAIL), serum IFN-γ and membrane-bound TRAIL expression on peripheral leucocytes from 58 CHB patients were examined by ELISA and flow cytometry respectively. The levels of TRAIL were compared with the baseline levels of 17 healthy controls, and correlation analysis was performed between ALT, TBIL, PT, morphological change in hepatic tissues, and serum IFN-γ. RESULTS: The results showed that TRAIL levels on membranes of CD4+, CD8+ T cells in CHB patients were much higher than those in healthy controls (P<0.001), and were correlated with serum TBIL (r=0.354, P= 0.008 for CD4+ and r= 0.522, P= 0.000 for CD8+, respectively), ALT (r= 0.393, P= 0.003 for CD8+), PT (r = 0.385, P = 0.004 for CD8+) and serum IFN-y level (r = 0.302, P= 0.011 for CD4+ and r= 0.307, P= 0.009 for CD8+). On the contrary to membrane-bound TRAIL expression, serum level of sTRAIL was not correlated with that of TBIL and PT, though it was higher than that of the normal population and was positively correlated with serum HBeAg expression (r= 0.695, P = 0.001). CONCLUSION: The expression level of TRAIL on the membrane of lymphocytes was upregulated and associated with the liver injury in CHB patients. These findings suggest that upregulation of TRAIL expression may be induced by virus antigen and inflammatory cytokine IFN-γ.

Induction of ischemic tolerance in rat liver via reduced nicotinamide adenine dinucleotide phosphate oxidase in Kupffer cells

AIM: To elucidate the mechanisms of hepatocyte preconditioning by H2O2 to better understand the pathophysiology of ischemic preconditioning. METHODS: The in vitro effect of H2O2 pretreatment was investigated in rat isolated hepatocytes subjected to anoxia/reoxygenation. Cell viability was assessed with propidium iodide fluorometry. In other experiments, rat livers were excised and subjected to warm ischemia/ reperfusion in an isolated perfused liver system to determine leakage of liver enzymes. Preconditioning was performed by H2O2 perfusion, or by stopping the perfusion for 10 min followed by 10 min of reperfusion. To inhibit Kupffer cell function or reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, gadolinium chloride was injected prior to liver excision, or diphenyleneiodonium, an inhibitor of NADPH oxidase, was added to the perfusate, respectively. Histological detection of oxygen radical formation in Kupffer cells was performed by perfusion with nitro blue tetrazolium. RESULTS: Anoxia/reoxygenation decreased hepatocyte viability compared to the controls. Pretreatment with H2O2 did not improve such hepatocyte injury. In liver perfusion experiments, however, H2O2 preconditioning reduced warm ischemia/reperfusion injury, which wasreversed by inhibition of Kupffer cell function or NADPH oxidase. Histological examination revealed that H2O2 preconditioning induced oxygen radical formation in Kupffer cells. NADPH oxidase inhibition also reversed hepatoprotection by ischemic preconditioning. CONCLUSION: H2O2 preconditioning protects hepato- cytes against warm ischemia/reperfusion injury via NADPH oxidase in Kupffer cells, and not directly. NADPH oxidase also mediates hepatoprotection by ischemic preconditioning.

Establishment of an animal model of ischemic type intrahepatic biliary lesion in rabbits

AIM:To explore a method to establish an animal model of ischemic type intrahepatic biliary lesion in rabbits. METHODS:Forty Japanese white rabbits of clean grade were divided randomly into four groups(10 rabbits per group)including sham operation(SO) group,and artery-bile obstruction(ABO)-1 h group, ABO-2 h group and ABO-3 h group.All the rabbits in this study underwent the same initial surgical procedure in which the liver was prepared as for graft removal during liver transplantation.Subsequently in the SO group,no additional vascular intervention was performed,while in groups ABO-1 h,ABO-2 h and ABO-3 h,the animals underwent combined clamping of the hepatic artery and common bile duct with microvascular clips for 1,2 and 3 h,respectively.After the scheduled occlusion time,the clip was removed to recover blood supply.The animals were killed 4 wk after operation.The survival rate,liver function, cholangiography and histopathological manifestation of the rabbits in each group were observed. RESULTS:The survival rate was 100%in groups SO,ABO-1 h and ABO-2 h,while it was 60%in group ABO-3 h.At each observation time,the change degree of the indexes of liver function was proportional to the clamping time(ABO-3 h>ABO-2 h>ABO-1 h> SO,P<0.05).Cholangiographical and histopathologic manifestations both showed that intrahepatic biliary lesion aggravated proportionally with the increase of the clamping time. CONCLUSION:An animal model of ischemic type intrahepatic biliary lesion in rabbits is successfully established,which may provide a reliable technique for basic and clinical research into the etiology, development and prophylaxis of ischemic type intrahepatic biliary lesion after liver transplantation.

Investigation on correlation between expression of CD58 molecule and severity of hepatitis B

AIM： To investigate the correlation between expression of CD58 and severity of hepatitis B. METHODS： The level of soluble CD58 （sCD58） in serum of patients with hepatitis B was detected by enzymelinked immunosorbent assay. The level of expression of membrane CD58 molecule in PBMC was detected by direct immunofluorescence. The levels of serumal TBIL, DBIL, IBIL, ALT and AST were detected by the automated biochemistry analyzer as well. RESULTS： The levels of sCD58 in serum and membrane CD58 molecule in PBMC of patients with hepatitis B were significantly higher than that in normal controls （P 〈 0.05）. Level of CD58 was related to the levels of serumal TBIL, DBIL, IBIL, ALT and AST. CONCLUSION： The level of CD58 molecule （in both serum and PBMC form） of patients with hepatitis B is related to the degree of liver damage.

Coexistence of hyperlipidemia and acute cerebral ischemia/reperfusion induces severe liver damage in a rat model

AIM:To investigate the correlation of hyperlipemia(HL) and acute cerebral ischemia/reperfusion(I/R) injury on liver damage and its mechanism.METHODS:Rats were divided into 4 groups:control,HL,I/R and HL+I/R.After the induction of HL via a high-fat diet for 18 wk,middle cerebral artery occlusion was followed by 24 h of reperfusion to capture I/R.Serum alanine transaminase(ALT) and aspartate aminotransferase(AST) were analyzed as part of liver function tests and liver damage was further assessed by histological examination.Hepatocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labeling(TUNEL) assay.The expression of genes related to apoptosis(caspase-3,bcl-2) was assayed by immunohistochemistry and Western blotting.Serum tumor necrosis factor-(TNF-),interleukin-1(IL-1) and liver mitochondrial superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),malondialdehyde(MDA) and Ca 2+ levels were measured to determine inflammatory and oxidative/antioxidative status respectively.Microsomal hydroxylase activity of the cytochrome P450 2E1(CYP2E1)-containing enzyme was measured with aniline as the substrate,and CYP2E1 expression in the liver tissue and microsome was determined by immunohistochemistry and Western blotting respectively.RESULTS:HL alone induced by high-fat diet for 18 wk resulted in liver damage,indicated by histopathological analysis,and a considerable increase in serum ALT(25.13 ± 16.90 vs 9.56 ± 1.99,P < 0.01) and AST levels(18.01 ± 10.00 vs 11.33 ± 4.17,P < 0.05) compared with control.Moreover,HL alone induced hepatocyte apoptosis,which was determined by increased TUNEL-positive cells(4.47 ± 0.45 vs 1.5 ± 0.22,P < 0.01),higher caspase-3 and lower bcl-2 expression.Interestingly,compared with those in control,HL or I/R groups,massive increases of serum ALT(93.62 ± 24.00 vs 9.56 ± 1.99,25.13 ± 16.90 or 12.93 ± 6.14,P < 0.01) and AST(82.32 ± 26.92 vs 11.33 ± 4.17,18.01 ± 10.00 or 14.00 ± 6.19,P < 0.01) levels in HL+I/R group were observed suggesting severe liver damage,which was confirmed by liver histology.In addition,HL combined with I/R also caused significantly increased hepatocyte apoptosis,as evidenced by increased TUNEL-positive cells(6.20 ± 0.29 vs 1.5 ± 0.22,4.47 ± 0.45 or 1.97 ± 0.47,P < 0.01),elevated expression of caspase-3 and lower expression of bcl-2.Furthermore,when compared to HL or I/R alone,HL plus I/R enhanced serum TNF-,IL-1,liver mitochondrial MDA and Ca 2+ levels,suppressed SOD and GSH-Px in liver mitochondria,and markedly up-regulated the activity(11.76 ± 2.36 vs 4.77 ± 2.31 or 3.11 ± 1.35,P < 0.01) and expression(3.24 ± 0.38 vs 1.98 ± 0.88 or 1.72 ± 0.58,P < 0.01) of CYP2E1 in liver.CONCLUSION:The coexistence of HL and acute cerebral I/R induces severe liver damage,suggesting that cerebral ischemic stroke would exaggerate the damage of liver caused by HL.This effect is possibly due to en-hanced CYP2E1 induction which further promotes oxidative damage,inflammation and hepatocyte apoptosis.

Ataxia telangiectasia-mutated-Rad3-related DNA damage checkpoint signaling pathway triggered by hepatitis B virus infection

AIM： To explore whether acute cellular DNA damage response is induced upon hepatitis B virus （HBV） infection and the effects of the HBV infection. METHODS： We incubated HL7702 hepatocytes with HBV-positive serum, mimicking a natural HBV infection process. We used immunoblotting to evaluate protein expression levels in HBV-infected cells or in non-infected cells; immunofluorescence to show ATR foci ands Chkl phosphorylation foci formation; flow cytometry to analyze the cell cycle and apoptosis; ultraviolet （UV） radiation and ionizing radiation （IR）-treated cells to mimic DNA damage; and Trypan blue staining to count the viable cells. RESULTS： We found that HBV infection induced an increased steady state of ATR protein and increased phosphorylation of multiple downstream targets including Chkl, p53 and H2AX. In contrast to ATR and its target, the phosphorylated form of ATM at Ser-1981 and its downstream substrate Chk2 phosphorylation at Thr-68 did not visibly increase upon infection. However, the level of Mre11 and p21 were reduced beginning at 0.5 h aEer HBV-positive serum addition. Also, HBV infection led to transient cell cycle arrest in the S and the G2 phases without accompanying increasedapoptosis. Research on cell survival changes upon radiation following HBV infection showed that survival of UV-treated host cells was greatly increased by HBV infection, owing to the reduced apoptosis. Meanwhile, survival of IR-treated host cells was reduced by HBV infection. CONCLUSION： HBV infection activates ATR DNA damage response to replication stress and abrogates the checkpoint signaling controlled by DNA damage response.

Effect of ONO-4057 and tacrolimus on ischemia-reperfusion injury of the liver

AIM: To investigate the effects of a novel Leukotriene B4 receptor antagonist and/or tacrolimus on ischemia-reperfusion in a rat liver model. METHODS: Male Lewis rats were pretreated with ONO-4057 (100 mg/kg) and/or tacrolimus (1 mg/kg) orally, and divided into four experimental groups; group 1 (control), group 2 (ONO-4057), group 3 (tacrolimus), group 4 (ONO-4057 + tacrolimus). RESULTS: There was a tendency for long survival in the groups treated with tacrolimus alone and ONO-4057 plus tacrolimus. Post-reperfusion serum aspartate aminotransferase levels decreased more signif icantly in ONO-4057 plus tacrolimus group (P < 0.01), than in the tacrolimus alone group (P < 0.05), compared to controls. CONCLUSION: This study demonstrated that pretreat-ment with ONO-4057 in combination with tacrolimus produced additive effects in a rat model of liver isch-emia-reperfusion injury.

Evaluation of CMU-1 preservation solutions using an isolated perfused rat liver model

AIM: CMU-1 is a new preservation solution with a low potassium concentration as well as low viscosity that is highly effective in reducing preservation injury. The purpose of this experiment is to compare the protective effect of CMU-1 solution with that of UW during cold preservation and normothermic reperfusion. METHODS: Wistar rats were divided into two groups according to different preservation solution: CMU-1 group and UW group. After 6, 12 and 24 h cold storage of rat liver in different preservation solutions, the isolated perfused rat liver model was applied to reperfuse the liver for 120 min normothermically (37 癈) with Krebs-Henseleit solution, meanwhile the pH value of the preservation solution was measured. The perfusate was sampled for the evaluation of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). At the end of the reperfusion, all of the bile product was collected, energy metabolic substrate and histological examination were performed. RESULTS: After preserving for 6 h, pH value of both groups did not change; after 12 h, both decreased but with no significant difference. After 24 h, pH value in UW solution group significantly decreased. The total adenine nucleotides level and AEC in liver tissue decreased with preservation time, but they were higher in CMU-1 group. And the amount of bile product after perfusion for 120 min in CMU-1 group was much more than that in UW group. However, there were no significant differences in ALT and LDH levels between two groups. Histology showed no difference. CONCLUSION: The preservation effect of CMU-1 solution is similar with that of UW solution. However, CMU-1 solution shows some advantages over UW solution in energy metabolism, preventing intracellular acidosis and bile product.

Oxidative damage,pro-inflammatory cytokines,TGF-αand c-myc in chronic HCV-related hepatitis and cirrhosis

AIM： To assess whether a correlation exists between oxidative DNA damage occurring in chronic HCV-related hepatitis and expression levels of pro-inflammatory cytokines, TGF-α and c-myc. METHODS： The series included 37 patients with chronic active HCV-related hepatitis and 11 with HCV-related compensated cirrhosis. Eight-hydroxydeoxyguanosine in liver biopsies was quantified using an electrochemical detector. The mRNA expression of TNF-α, IL-1β, TGF-α and c-myc in liver specimens was detected by semiquantitative comparaUve RT-PCR. RESULTS： TNF-α levels were significantly higher in hepatitis patients than in cirrhosis patients （P=0.05）. IL-1β was higher in cirrhosis patients （P=0.05）. A sig- nificant correlation was found between TNF-α and staging （P= 0.05） and between IL-1β levels and grading （P=0.04）. c-myc showed a significantly higher expression in cirrhosis patients （P=0.001）. Eight-hydroxydeoxyguanosine levels were significantly higher in cirrhosis patients （P=0.05） and in HCV genotype 1 （P=0.03）. Considering all patients, 8-hydroxydeoxyguanosine levels were found to be correlated with genotype （P=0.04） and grading （P=0.007）. Also multiple logistic regression analysis demonstrated a significant correlation among the number of DNA adducts, TNF-α expression and HCV genotype （P= 0.02）. CONCLUSION： In chronic HCV-related liver damage, oxidative DNA damage correlates with HCV genotype, grading and TNF-α levels. As HCV-related liver damage progresses, TNF-α levels drop while IL-1β and c-myc levels increase, which may be relevant to liver carcinogenesis.

Expression of nitric oxide synthase in T-cell-dependent liver injury initiated by ConA in Kunming mice

Objective: To investigate whether nitric oxide synthase (NOS) is expressed in T-cell-dependent liver injury initiated by concanavalin A (ConA) in Kunming mice and study the possible effect of nitric oxide(NO) on liver injury models. Methods: Liver injury in Kunming mice was induced by administration of ConA through tail vein. Expression of NOS in the liver was detected by NADPH diaphorase staining method. The possible effect of NO on liver injury models was obtained by L-NAME injection to suppress synthesis of NO. Results: NOS has a strong expression in hepatocytes after ConA injection, especially in those close to the central vein, while only a weak expression was found in the epithelial cells in control group. Liver injury became more serious when NO synthesis was inhibited by L-NAME, accompanied by great malondialdehyde(MDA) increase in serum and severe intrahepatic vascular thrombosis. Conclusion: NOS markedly expressed in ConA-induced liver injury, which may subsequently promote nitric oxide synthesis. Increasement of nitric oxide has a protective effect on ConA-induced liver injury.

Over-expression of uPA increases risk of liver injury in pAAV-HBV transfected mice

AIM:To investigate the relationship between overexpression of urokinase plasminogen activator(uPA) and hepatitis B virus(HBV) related liver diseases in a transgenic mouse model.METHODS:Albumin-tetracycline reverse transcriptional activator and tetO-uPA transgenic mice were generated respectively through pronuclear injection and crossed to produce the double transgenic in-alb-uPA mice,for which doxycycline(Dox)-inducible and liver-specific over-expression of uPA can be achieved.Hydrodynamic transfection of plasmid adeno-associated virus(AAV)1.3HBV was performed through the tail veins of the Dox-induced in-alb-uPA mice.Expression of uPA and HBV antigens were analyzed through double-staining immunohistochemical assay.Cytokine production was detected by enzyme linked immunosorbent assay and α-fetoprotein(AFP) mRNA level was evaluated through real-time quantitative polymerase chain reaction.RESULTS:Plasmid AAV-1.3HBV hydrodynamic transfection in Dox-induced transgenic mice not only resulted in severe liver injury with hepatocarcinoma-like histological changes and hepatic AFP production,but also showed an increased serum level of HBV antigens and cytokines like interleukin-6 and tumor necrosis factor-α,compared with the control group.CONCLUSION:Over-expression of uPA plays a synergistic role in the development of liver injury,inflammation and regeneration during acute HBV infection.

Acute sensorineural hearing loss associated with peginterferon and ribavirin combination therapy during hepatitis C treatment: Outcome after resumption of therapy

Peginterferon and ribavirin combination therapy for the treatment of hepatitis C virus （HCV） is well known to be associated with significant adverse effects. Sensorineural hearing loss, that in most cases is unilateral, has been reported as a consequence of therapy with both non-pegylabed and pegylated interferon （pegIFN） but is not a well-known adverse effect. We report a 45-year-old Caucasian woman who developed acute sensorineural hearing loss 2 mo after starting therapy with pegIFN-α 2b and ribavirin for the treatment of chronic HCV, genotype la. She did not report the hearing loss to the hepatitis clinic until 1 mo, later whereupon therapy was promptly discontinued. Although her serum alanine aminotransferase （ALT） normalized and her HCV-RNA became undetectable after 12 wk of pegIFN and dbavirin therapy, after discontinuation, her HCV-RNA became detectable with significant elevations of serum ALT. Four months after initial discontinuation, the patient re-commenced pegIFN and ribavirin combination therapy. After 44 of 48 wk of therapy, the patient＇s liver biochemistry has normalized and the HCV-RNA is undetectable. She has not developed worsening of her hearing loss and hearing on the left-side is unaffected. Both patients and physicians should be aware that sensorineural hearing loss may occur with pegIFN therapy. Our experience suggests that re-institution of therapy is not always associated with further hearing impairment.

Laser capture microdissection and genetic analysis of carbon-labeled Kupffer cells

AIM： To develop a method of labeling and microdissecting mouse Kupffer cells within an extraordinarily short period of time using laser capture microdissection （LCM）. METHODS： Tissues are complex structures comprised of a heterogeneous population of interconnected cells. LCM offers a method of isolating a single cell type from specific regions of a tissue section. LCM is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCM prior to mRNA isolation can be elaborate, thereby subjecting the RNA to considerable degradation. Kupffer cell labeling is achieved by injecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model. RESULTS： mRNA extracted from the microdissected cell population displayed marked increases in colonystimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. Gene expression by Kupffer ceils derived from bile-duct-ligated, versus sham-operated, mice was compared. Microarray analysis revealed a significant （2.5-fold, q value 〈 10） change in 493 genes. Based on this fold-change and a standardized PubMed search, 10 genes were identified that were relevant to the ability of Kupffer cells to suppress liver injury. CONCLUSION; The methodology outlined herein provides an approach to isolating high quality RNA from Kupffer cells, without altering the tissue integrity.

Pathological and MR-DWI study of the acute hepatic injury model after stem cell transplantation

AIM： To investigate apparent diffusion coefficient （ADC） values as an indication of reconditioning of acute hepatic injury （AHI） after allogeneic mononuclear bone marrow cell （MBMC） transplantation. METHODS： Three groups were used in our study： a cell transplantation group （n = 21）, transplantation control group （n = 21） and normal control group （n = 10）. AHI model rabbits in the cell transplantation group were injected with 5 mL of MBMC suspension at multiple sites in the liver and the transplantation controls were injected with 5 mL D-Hanks solution. At the end of the 1st, 2nd and 4th wk, 7 rabbits were randomly selected from the cell transplantation group and trans- plantation control group for magnetic resonance diffu- sion-weighted imaging （MR-DWI） and measurement of the mean ADC values of injured livers. After MR-DWI examination, the rabbits were sacrificed and the livers subjected to pathological examination. Ten healthy rab- bits from the normal control group were used for MR- DWI examination and measurement of the mean ADC value of normal liver. RESULTS： At all time points, the liver pathological scores from the cell transplantation group were significantly lower than those in the transplantation control group （27.14± 1.46 vs 69.29± 6.16, 22.29 ± 2.29 vs 57.00± 1.53, 19.00 ± 2.31 vs 51.86 ± 6.04, P = 0.000）. The mean ADC values of the cell transplantation group were significantly higher than the transplantation con- trol group （（1.07± 0.07） ×10^-3 mm2/s vs （0.69 ± 0.05） ×10^-3 mm2/s, （1.41± 0.04） ×10^-3 mm2/s vs （0.84± 0.03） ×10^-3 mm2/s, （1.68 ± 0.04） ×10^-3 mm2/s vs （0.86± 0.04） ×10^-3 mm2/s, P = 0.000）. The pathological scores of the cell transplantation group and transplantation control group gradually decreased. However, their mean ADC values gradually increased to near that of the normal control. At the end of the 1st wk, the mean ADC values of the cell transplantation group and transplantation control group were significantly lower than those of the normal control group [（1.07 ± 0.07） ×10^-3 mm2/s vs （± 0.03） ×10^-3 mm2/s, （0.69± 0.05） ×10^-3 mm2/s vs （1.76 ± 0.03） ×10^-3 mm2/s, P = 0.000]. At any 2 time points, the pathological scores and the mean ADC values of the cell transplantation group were significantly different （P = 0.000）. At the end of the 1st wk, the pathological scores and the mean ADC values of the transplantation control group were significantly different from those at the end of the 2nd and 4th wk （P = 0.000）. However, there was no significant difference between the 2nd and 4th wk （P = 0.073 and 0.473, respectively）. The coefficient of correlation between the pathological score and the mean ADC value in the cell transplantation group was -0.883 （P = 0.000） and -0.762 （P = 0.000） in the transplantation control group. CONCLUSION： Tracking the longitudinally dynamic change in the mean ADC value of the AHI liver may reflect hepatic injury reconditioning after allogeneic MBMC transplantation.

Alleviation of ischemia/reperfusion injury in ob/ob mice by inhibiting UCP-2 expression in fatty liver

AIM： To investigate the protective effect of target suppression of uncoupling protein-2 （UCP-2） on ischemia/ reperfusion （I/R） injury in fatty liver in ob/ob mice. METHODS： Plasmids suppressing UCP-2 expression were constructed, and transfected into fatty liver cells cultured in vitro and the ob/ob mouse I/R injury model. Serum tumor necrosis factor （TNF）-α levels, UCP-2 mRNA expression, alanine aminotransferase （ALT） levels in ob/ob mice were tested, and the pathological changes in fatty liver were observed in experimental and control groups. RESULTS： In ob/ob mouse I/R models, serum TNF-α levels were significantly higher than in normal controls. After the plasmids were transfected into the cultured cells and animal models, expression of UCP-2 rnRNA was significantly reduced as compared with that in the control group （2^1.56± 0.15 vs 2^-0.45± 0.15, p 〈 0.05）. In ob/ob mouse models, in which expression of UCP-2 was suppressed, serum ALT levels were significantly lower than those of other groups, and pathological analysis revealed that injury of liver tissues was significantly alleviated. CONCLUSION： The target suppression of UCP-2 expression in fatty liver can alleviate the I/R injury in the ob/ob mice.

Serum levels of microRNAs can specifically predict liver injury of chronic hepatitis B

AIM： To investigate whether circulating microRNAs （miRNAs） can serve as molecular markers to predict liver injury resulted from chronic hepatitis B （CHB）. METHODS： The profiles of serum miRNA expression were first generated with serum samples collected from 10 patients with CHB and 10 healthy donors （Ctrls） by microarray analysis. The levels of several miRNAs were further quantitated by real-time reverse transcription polymerase chain reaction with serum samples from another 24 CHB patients and 24 Ctrls. Serum samples of 20 patients with nonalcohlic steatohepatitis （NASH） were also included for comparison. The comparison in the levels of miRNAs between groups （CHB, NASH and Ctrl） was analyzed with Mann-Whitney U-test. The cor- relation between miRNAs and clinical pathoparameters was analyzed using Spearman correlation analysis or canonical correlation analysis. The receiver-operator characteristic （ROC） curves were also generated to de- termine the specificity and sensitivity of each individual miRNA in distinguishing patients with CriB from Ctrls. RESULTS： miRNA profile analysis showed that 34 miR- NAs were differentially expressed between CriB and Ctrl subjects, in which 12 were up-regulated and 22 down-regulated in CriB subject （fold change 〉 2.0 and P 〈 0.01）. The median levels of miR-122, -572, -575 and -638 were significantly higher （P 〈 1.00 × 10-5） while miR-744 significantly lower （P 〈 1.0× 10-6） in Crib compared with the Ctrl. The levels of miR-122, -572 and -638 were also higher （P 〈 1.00×10-3） while the level of miR-744 lower in CriB （P 〈 0.05） than in NASH, although the difference between them was not as significant as that between CHB and Ctrl. ROC curve analysis revealed that the levels of miR-122, -572, -575, -638 and -744 in serum were sensitive and specific enough to distinguish CriB, NASH and Ctrl. Multivariate analysis further showed that the levels of these miRNAs were correlated with the liver function parameters. Most significantly, it was the scatter plot of principal component with the levels of these miRNAs, but not the parameters of liver function, which clearly distinguished CriB, NASH and Ctrl subjects. CONCLUSION： Serum levels of miR-122, -572, -575, -638 and -744 are deregulated in patients with CHB or NASH. The levels of these miRNAs may serve as po- tential biomarkers for liver injury caused by CHB and NASH.