2007年·第120卷第24期

Hepatitis B virus (HBV) infection is a worldwide health problem.According to WHO estimation,about 2 billion people globally have been infected With HBV and 350 million of them have chronic HBV infection.1

Chronic hepatitis B is one of the most common epidemic diseases in China and has become a major health issue.To help standardize the prevention,diagnosis,and treatment of chronic hepatitis B,the Guideline on prevention and treatment of chronic hepatitis B (abbr.Guideline) was created by a group of appropriate experts belonging to the Society of Hepatology and the Society of Infectious Disease,the Chinese Medical Association according to the principles of evidence-based medicine using the latest clinical research data。

Abstract：Background Hepatitis C virus(HCV)envelope genes encoding glycoproteins E1 and E2 exhibits a high degree of variability that gives rise to differing phenotypic traits;including alterations in receptor-binding affinity and immune recognition and escape.This study aims to elucidate the relationship of the evolutionary patterns for HCV envelope glycoproteins to viral persistence.Methods HCV quasispecies were characterized in specimens collected every two to six months from a cohort of acutely HCV-infected subjects.We evaluated two individuals who spontaneously cleared viremia and three individuals with persistent viremia by cloning 33 1-kb amplicons that spanned E1 and the 5'half of E2;including hypervariable region 1(HVR1).To detect representative variants for sequencing thirty-three cloned cDNAs representing each specimen were assessed by a method that combined analysis of a single-stranded conformational polymorphism(SSCP)method and heteroduplex analysis(HDA).For each patient,the rates of both synonymous and nonsynonymous substitutions for the E1,HVR1 and E2 regions outside HVR1 were evaluated.The amino acid sequences and predicted antigenic profiles were analyzed.Results The genetic diversity within HVR1 was consistently higher than that in the E1 and E2 regions outside HVR1 in individuals with persistent viremia,but did not change markedly over time in those with clearance of viremia.For individuals with persistent viremia,the rate of nonsynonymous substitutions within the HVR1 region predominated and gradually increased,compared to that in the E1 and E2 regions outside HVR1.By contrast,the rates of both nonsynonymous and synonymous substitutions for the E1 and E2 regions,including HVR1,were consistently lower in individuals with clearance of viremia.HVR1 had a higher antigenic variable and lower positive charge in subjects with persistent viremia.All cysteine residues and N-linked glycosylation sites,some of which were known to play a major role in protein folding and others play a role in HCV entry,were 100% conserved among the sequenced cloned cDNAs from the two outcome groups.Conclusion HCV persistence may be associated with positive selection pressures on HVR1,rather than functional constraints in the envelope region。

Abstract：Background Hepatitis B virus(HBV)X protein(HBx)and p53 could mutually down-regulate at transcriptional level and HBx could bind with p53 protein within its transactivation domain and inhibit the function of p53 protein.In recent years,effects of arsenic trioxide(As2O3)on the expression of p53 protein have been widely studied,while little is known about the activity of p53 protein.This study was undertaken to delineate the effect of HBV X gene and As2O3 on p53 protein expression(level and activity)in HepG2 cells by small hairpin RNA(shRNA)-mediated RNA interference(RNAi)technique.Methods Cell line HepG2 and cells with stable expression of HBV X gene(HepG2-X)were treated with 2 μmol/L As2O3,with corresponding untreated cells serving as controls.Cell lysates and nuclear extracts were extracted.Total level and the relative activity of p53 protein were detected by modified enzyme-linked immunosorbent assay(ELISA).HBV X gene sequence-specific shRNA expression vector(pXi-1 and pXi-2)and sequence-unrelated control(pXi-3)were transfected into HepG2-X.Single cell clone with stable expression of shRNA was selected and exposed to propagating culture.The effect of As2O3 on p53 protein expression and activity was re-observed.Results Total p53 protein level was up-regulated and its relative activity ratio was enhanced by As2O3 in HepG2 and HepG2-X cells.The total p53 protein level induced by As2O3 was up-regulated by HBV X gene expression,while its relative activity was significantly suppressed.The suppression was removed after HBV X gene expression was repressed by shRNA.Conclusions As2O3 up-regulates p53 protein expression and enhance its activity.HBV X up-regulates As2O3 induced-p53 protein expression while suppresses its activity。

Abstract：Background Low-density lipoprotein(LDL)receptor is normally regulated via a feedback system that is dependent on intracellular cholesterol levels.We have demonstrated that cytokines disrupt cholesterol-mediated LDL receptor feedback regulation causing intracellular accumulation of unmodified LDL in peripheral cells.Liver is the centraI organ for lipid homeostasis.The aim of this study was to investigate the regulation of cholesterol exogenous uptake via LDL receptor and its underlying mechanisms in human hepatic cell line(HepG2)cells under physiological and inflammatory conditions.Methods Intracellular total cholesterol(TC),free cholesterol(FC)and cholesterol ester(CE)were measured by an enzymic assay.Oil Red O staining was used to visualize lipid droplet accumulation in cells.Total cellular RNA was isolated from cells for detecting LDL receptor,sterol regulatory element binding protein (SREBP)-2 and SREBP cleavage-activating protein(SCAP)mRNA levels using real-time quantitative PCR.LDL receptor and SREBP-2 protein expression were examined by Western blotting.Confocal microscopy was used to investigate the translocation of SCAP-SREBP complex from the endoplasmic reticulum(ER)to the Golgi by dual staining with anti-human SCAP and anti-Golgin antibodies.Results LDL loading increased intracellular cholesterol level,thereby reduced LDL receptor mRNA and protein expression in HepG2 cells under physiological conditions.However,interleukin 1β(IL-1β)further increased intracellular cholesterol level in the presence of LDL by increasing both LDL receptor mRNA and protein expression in HepG2.LDL also reduced the SREBP and SCAP mRNA level under physiological conditions.Exposure to IL-1β caused Over-expression of SREBP-2 and also disrupted normal distribution of SCAP-SREBP complex in HepG2 by enhancing translocation of SCAP-SREBP from the ER to the Golgi despite a high concentration of LDL in the culture medium.Conclusions IL-1β disrupts cholesterol-mediated LDL receptor feedback regulation by enhancing SCAP-SREBP complex translocation from the ER to the Golgi,thereby increasing SREBP-2 mediated LDL receptor expression even in the presence of high concentration of LDL.This results in LDL cholesterol accumulation in hepatic cells via LDL receptor pathway under inflammatory stress。

Abstract：Background Over 70% of the total tissue weight in the cartilage matrix consists of water,and the early-stage osteoarthritic cartilage is characterized by swelling.Water transport in the cartilage matrix and across the membranes of chondrocytes may be important in normal and pathological conditions of cartilage.The purpose of this study was to identify aquaporin-1 (AQP1) and aquaporin-3 (AQP3) expressions in the mandibular condylar cartilage after experimentally induced osteoarthritis(OA)in rats.Methods An experimental temporomandibular joint OA was induced by partial discectomy in rats.The pathological characteristics of the normal,early-stage,and late-stage osteoarthritic TMJ cartilages were verified by histological techniques.The AQP1 and AQP3 gene expressions in the normal and osteoarthritic cartilages were measured using quantitative real-time reverse-transcription PCR analysis.The cartilage sections were incubated in primary polyclonal antibodies to AQP3;immunofluorescent microscopy was used to examine the AQP3 expression shown by its protein level.Results The mRNA expression levels of AQP1 and AQP3,analyzed using quantitative PCR,revealed that AQP3 mRNA was highly up-regulated in the OA cartilage,which was considered significant.There was no notable difference in the expression of AQP1 mRNA between OA and normal controls.With the progressing of the OA,the localization of the AQP3 protein was quite different from that of the normal cartilage.Cormpared to the normal cartilage,the expressions of AQP3 protein were observed mainly in the proliferative zone and the upper mid-zone chondrocytes at the early-stage of OA,and were observed to appear frequently throughout the mid-and deep zone during the late-stage of OA.Conclusions The high expression of AQP3 mRNA in the OA cartilage and the different localization of the AQP3 protein suggest that it may play a particular role in OA pathogenesis.Further study of AQP3 function may provide new insight into the understanding of the molecular mechanisms underlying OA。

Abstract：Background Although severe acute respiratory syndrome(SARS)has been controlled,the subsequently emerging sporadic cases in 2004 emphasize the necessity of developing a rapid diagnostic method,which would be of great help in clinical diagosis and also wild host screening.This study aims to establish an effective and rapid serological tool for the diagnosis of SARS-CoV by comparison among whole viral,N and N199 proteins by ELISA.Methods SARS-CoV N and N199(a truncated nucleocapsid gene)genes were cloned,expressed,identified by Western blotting,and applied in screening of human and swine samples.Sera of SARS convalescent-phase patients,normal human sera,sera of patients with other respiratory diseases,and swine sera were screened by ELISA,with whole SARS-CoV F69,N and N199 proteins as antigens.Results The sensitivity and specificity of N and N199 proteins in human sera diagnosis were approximate(P=0.743),which was higher than whole viral protein but the difference was not significant(P=0.234).The N199 protein proved to be more specific in swine sera screening than whole viral and N protein(P<0.001).Conclusion N199 protein is feasible in both clinical diagnosis and SARS-CoV reservoir screening。

Abstract：Background Pittsburgh modified TNM criteria is one of the prognostic models of orthotopic liver transplantation(OLT) for hepatocellular carcinoma(HCC).In this study,we applied this prognostic system in a series of HCC patients receiving OLT to verify its reliability in the clinical prognostic prediction.Methods The clinical record and follow-up data of 102 patients with HCC underwent OLT was collected.The patients were classified by 3 staging systems:the Pittsburgh Modified TNM Criteria,International Union Against Cancer(UICC) pTNM Staging System,and Milan Criteria.Survival rates of the patients were analyzed using the Kaplan-Meier method and the Log-Rank test,and then the prognostic values of the 3 staging systems were compared.Results Among the 3 staging systems,the Pittsburgh Modified TNM Criteria showed the best stratification of patients with different prognosis.The overall survival rates of the patients at the Pittsburgh modified TNM stage Ⅰ,Ⅱ,Ⅲ,and IV were 94.4%,83.3%,58.2%,and 36.8% at 1 year,and 79.4%,62.5%,26.2%,and 10.5% at 3 years,respectively.For those patients exceeding the Milan Criteria,the patients at Pittsburgh stages Ⅰ and Ⅱ had a significant higher survival rate than those at Pittsburgh stages Ⅲ and Ⅳ (P<0.001).Conclusions The Pittsburgh Modified TNM Criteria is a more reliable postoperative staging system than the UICC pTNM staging system for HCC patients receiving OLT.As providing more accurate prognostic classification,it could be reasonable to combine the Milan Criteria for recipient selection。

Abstract：Background Although severe encephalopathy has been proposed as a possible contraindication to the use of noninvasive positive-pressure ventilation(NPPV),increasing clinical reports showed it was effective in patients with impaired consciousness and even coma secondary to acute respiratory failure,especially hypercapnic acute respiratory failure(HARF).To further evaluate the effectiveness and safety of NPPV for severe hypercapnic encephalopathy,a prospective case-control study was conducted at a university respiratory intensive care unit(RICU)in patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)during the past 3 years.Methods Forty-three of 68 consecutive AECOPD patients requiring ventilatory support for HARF were divided into 2 groups,which were carefully matched for age,sex,COPD course,tobacco use and previous hospitalization history,according to the severity of encephalopathy,22 patients with Glasgow coma scale(GCS)<10 served as group A and 21 with GCS≥10 as group B.Results Compared with group B,group A had a higher level of baseline arterial partial CO2 pressure((102±27)mmHg vs (74±17)mmHg,P<0.01),lower levels of GCS (7.5±1.9 vs 12.2±1.8,P<0.01),arterial pH value(7.18±0.06 vs 7.28±0.07,P<0.01)and partial O2 pressure/fraction of inspired O2 ratio(168±39 vs 189±33,P<0.05).The NPPV success rate and hospital mortality were 73%(16/22)and 14%(3/22)respectively in group A,which were comparable to those in group B(68%(15/21)and 14%(3/21) respectively,all P>0.05),but group A needed an average of 7 cmH2O higher of maximal pressure support during NPPV,and 4,4 and 7 days longer of NPPV time,RICU stay and hospital stay respectively than group B(P<0.05 or P<0.01).NPPV therapy failed in 12 patients(6 in each group)because of excessive airway secretions(7 patients),hemodynamic instability(2),worsening of dyspnea and deterioration of gas exchange(2),and gastric content aspiration(1).Conclusions Selected patients with severe hypercapnic encephalopathy secondary to HARF can be treated as effectively and safely with NPPV as awake patients with HARF due to AECOPD;a trial of NPPV should be instituted to reduce the need of endotracheal intubation in patients with severe hypercapnic encephalopathy who are otherwise good candidates for NPPV due to AECOPD。

Abstract：Background Endovascular stent-grafting is widely used to treat thoracic aortic dissection.However,little information is available regarding outcome following simultaneous exclusion of multiple tears.This report details eight years of experience using simultaneous multi-tear exclusion for treatment of Stanford type B thoracic aortic dissection resulting in successful aortic remodeling without adverse events.Methods From September 1998 to January 2006,29 type B thoracic aortic dissection patients(24 men,5 women;27 chronic,2 acute;mean age 58 years,range 45-77 years)were treated by simultaneous multi-tear exclusion in our center.Magnetic resonance angiography was used as the preoperative evaluation method.Different kinds of stent-grafts were used.The patients were followed up with contrast-enhanced spiral computed tomography at 6 months postoperatively and yearly thereafter.Results Twenty-nine surgeries were completed successfully using at least 2 stent-grafts per patient(range:2-6,mean:2.7).No major procedure-related complications,such as rupture,paraplegia,aortic branch ischemia or cerebral infarction,were observed.During follow-up,favorable remodeling of the aorta was observed.Conclusions The mid-term result of thoracic aortic dissection with simultaneous multi-tear exclusion was satisfactory.With the improvement of stent-grafts,simultaneous multi-tear exclusion should find wider application and become an optimal strategy for thoracic aortic dissection。

Abstract：Background Despite recent advances in recognition and treatment of primary palmar hyperhidrosis(PPH),the epidemiological survey has hardly been conducted.The aim of this study was to i.lvestigate the prevalence and epidemiological characteristics of primary PPH among adolescents in three cities of southeast China.Methods Stratified-cluster sampling was carried out and cross-sectional epidemiological survey by questionnaire was applied among 33 000 college and high schooJ students.Results The prevalence rate of PPH was 4.36% affecting both sexes equally.Prevalence rate of severe PPH was 0.27%.The average age of onset was 12.27±2.12 years.The peak age of onset was 6-16 years,accounting for 97.2% of PPH population.Positive family history was found in 17.9% PPH cases.Besides palms,axillae and soles can be also affected.Conclusions PPH affects a larger group of individual than previously reported.More measures should be taken to enhance the recognition,diagnosis,and treatment of PPH。

Abstract：Background Calpain-10(CAPN10)has been identified as a susceptibility gene in type 2 diabetes mellitus (T2DM) and insulin resistance.The present study aimed to identify the effects of genetic variations in the CAPN10 gene on the development of type 2 diabetes and hypertension in northern Han Chinese population.Methods We performed a case-control study and genotyped single nucleotide polymorphism(SNP)-44,-43,-19 and -63 of CAPN10 gene in 1046 subjects from the northern China,including 493 patients with T2DM and hypertension and 553 age-and gender-matched normal healthy controls.Results Univariate analysis showed that the four polymorphisms were not independently associated with T2DM and hypertension.However,the frequency distributions of SNP-44 allele C(allele 2)(17.89% vs 9.80%,P=0.0016)and genotype CC(22)(4.21% vs 1.01%,P=0.0059)in obese patients(body mass index≥30 kg/m2)were different from those in non-obese patients.Logistic regression analyses revealed that carriers of lhe 1112/1221 diplotype had a significantly lower odds ratio for diabetes and hypertension(OR=0.399,95% CI,0.196-0.814,P=0.0115).The 1112/1121 diplotype associated with significantly increased risk of type 2 diabetes in Mexican-American was not associated with the increased risk in Chinese.Conclusion These results suggested that CAPN10 gene variations might play roles in the risk of diabetes and hypertension in northern Han Chinese population。

Abstract：Background Stimulation of the heart β3-adrenoceptor(AR)may result in a negative inotropic effect.Being up-regulated,β3-AR plays a more important role in the regulation of cardiac function during heart failure.However,the effect of chronic blocking of β3-AR on heart failure has not been fully elucidated.In this study,we used a selective β3-AR antagonist SR59230A to treat a well defined heart failure rat model chronically,then evaluated its effect on cardiac function and investigated the mechanism.Methods Male Wistar rats were chosen randomly as controls (n=8).Isoproterenol induced heart failure rats were randomly divided into ISO group(n=10) and SR group(n=10).The ISO group received intraperitoneal injection of 1 ml saline twice a day;the SR group received intraperitoneal injection of SR59230A 85 nmol in 1 ml saline twice a day;and the controI group received no treatment.The treatment was started 24 hours after the last isoproterenol injection and continued for 7 weeks.Then we measured the following indexes:the ratio of heart weight to body weight(HW/BW) and the ratio of left ventricular weight to body weight(LVW/BW),collagen volume fraction(CVF),left ventricular end diastolic dimension(LVEDd),left ventricular end systolic dimension(LVESd),ejection fraction(EF),fractional shortening(FS)and the ratio of E wave to A wave(E/A),the mRNA and protein expression of β3-AR and eNOS,and cGMP level in the heart.Results The ratios HW/BW and LVW/BW were significantly increased in the ISO group compared with the control group(P<0.01),but they were limited in the SR group(P<0.05 compared with the ISO group).CVF increased in the ISO group and the SR group(P<0.01),but it was significantly attenuated in the SR group(P<0.01).LVESd,LVESd and E/A ratio were significantly increased in the ISO group compared with the control group(P<0.01),while EF and FS were significantly decreased(P<0.01).Compared with the ISO group,the SR group showed that LVEDd,LVESd and E/A ratio were significantly decreased(P<0.01),whereas EF and FS were significantly increased(P<0.01).β3-AR and eNOS mRNA and protein in the ISO group were significantly increased when compared with the control group(P<0.01).These increases were all attenuated in the SR group compared with the ISO group(P<0.01).The level of cGMP in myocardial tissue was significantly increased in the ISO group compared with the control group(P<0.01),whereas SR59230A treatment normalized this increment(P<0.01).Conclusions Chronic blocking of β3-AR could ameliorate cardiac function in heart failure rats and its mechanism involves inhibition of the negative inotropic effect and attenuation of cardiac remodeling。

The liver plays a major role in the regulation of glucose,lipid and energy metabolism.Increased hepatic fat deposit is a very common feature in obese,insulin-resistant patients, in metabolic syndrome,alcoholic steatohepatitis (ASH) and nonalchoholic fatty liver disaseas(NAFLD)。

Abstract：Obiective To review the biological behaviour of endothelial progenitor cells and their role in vascular diseases.Data sources The data used in this review were mainly from Medline and PubMed for relevant English language articles published from 1985 to March 2007.The search term was "endothelial progenitor cells".Study selection Articles about the biological behaviour of endothelial progenitor cells and their roles in the pathogenesis of vascular diseases such as atherogenesis were used.Results Progenitor cells in bone marrow,peripheral blood and adventitia can differentiate into mature endothelial cells (ECs).The progenitor cells,which express certain surface markers including AC133,CD34 and KDR,enable restoration of the microcirculation and ECs when injury or ischaemia occurs.Endothelial progenitor cells used in experimental models and clinical trials for ischaemic syndromes could restore endothelial integrity and inhibit neointima development.Moreover,their number and functional properties are influenced by certain cytokines and atherosclerotic risk factors.Impairment of the progenitor cells might limit the regenerative capacity,even lead to the development of atherosclerosis or other vascular diseases.Conclusions Endothelial progenitor cells have a particular role in prevention and treatment of certain cardiovascular diseases.However,many challenges remain in understanding differentiation of endothelial progenitor cells,their mobilization and revascularization。

Hepatocellular carcinoma(HCC)is among the most common tumors worldwide.Despite extensive exploration of novel therapies,the prognosis of HCC patients with intrahepatic metastasis is still poor,and no treatment is effective against metastases.Therefore,more effective therapeutic strategies for treatment of metastasis are urgently needed。

Obstructive sleep apnea syndrome (OSAS) was a disease of breath obstacle happened in the process of sleep.The central mechanism of OSAS has not yet been fully elucidated.1

Pregnancy complicating fulminant hepatitis is characterized by rapid progress,complicated clinical symptoms,and association with blood coagulation dysfunction,hepatic encephalopathy,hepatorenal syndrome,toxic enteroparalysis and infection。

Since its introduction in 1995,laparoscopic living donor nephrectomy (LDN) has been shown to alleviate postoperative pain,shorten hospital stay,reduce loss and improve cosmesis while hastening the recovery of normal activities of donors.1-3 However,there has been much controversy over minimally invasive surgery with transperitoneal or retroperitoneal approach for nephrectomy.In this non-randomized retrospective study,we analyzed some parameters of the donors who had undergone transperitoneal LDN compared with those who had had retroperitoneoscopic nephrectomy。

Life expectancy in China doubled from 35 to 71 years between 1949 and 2001.China's human development index has been rising steadily,and it ranked 85th among countries with an index of 0.755 in 2003。

Coronary embolism has been considered as a rare cause of coronary occlusion.Prosthetic heart valve is an infrequent cause of this condition.The exact incidence of coronary embolization in patients with prosthetic heart valves is not well known。

The most common tumors associated with cardiac metastases are lung carcinoma and melanoma.1 Although the cardiac involvement of lung cancer is common(lung cancer usually directly invades into the pericardium and/or left/right atrium),the primary lung cancer accompanied with metastasis to the left ventricle is extremely rare。

Lung autotransplantation is a new procedure with pulmonary preservation for patients in the treatment of lung cancer.It was used to treat the stage Ⅲ central lung cancer of the upper lobe in the patient presented herein when his pulmonary function could not withstand a pneumonectomy and the length of the resected bronchus or pulmonary artery was too long to perform a bronchovascular double sleeve lobectomy.1,2 In order to preserve pulmonary tissue and improve the quality of life,we performed lung autotransplantation for a case。

Desmoplastic small round cell tumor(DSRCT)is a clinically and morphologically well-defined neoplasm.1 This highly aggressive malignant small cell neoplasm tends to affect adolescents and young adults and occurs predominantly in the abdomen,pelvis,and omentum.1,2 DSRCT in the lung is extremely rare。

Spontaneous pneumomediastinum (SPM),reported firstly by Hamman in 1939,is an uncommon,and usually benign,and self-limiting clinical disorder found in young people often without apparent precipitating factors or diseases。

Pulmonary actinomycosis is a rare, uncommon pulmonary infection.The word "actinomycosis" is derived from Greek terms aktino,which refers to the radiating appearance of a sulphur granule and myKos,which labels the condition a mycotic disease。

Facial nerve schwannoma involving the middle cranial fossa is quite rare,and its accurate diagnosis is very difficult before SUrgery.Here we present a case of schwannoma of the facial nerve at the middle cranial fossa that was misdiagnosed previously at a local hospital and then cured in our hospital。

Virus infection and subsequent myocardial damage and repairing were regarded as important aspects in the pathogenesis of dilated cardiomyopathy.…… Biography：Correspondence to:Dr. WANG Hong-yue,Department of Pathology,Fuwai Hospital and Cardiovascular Institute of Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,China(Tel:86-10-88398007.Email:hongyue_w@yahoo.com)

Intracranial arachnoid cysts,which often occur in children(75%),account for 1% of intracranial masses.1 The cyst is most commonly located at the middle cranial fossa and more frequently on the left side。

Pituitary apoplexy is a rare clinical syndrome caused by acute enlargement of pituitary adenomas,which may be secreting or nonfunctioning,and symptomatic or asymptomatic,resulted from hemorrhage or infarction.1