Objective To study the characteristics of lamivudine-resistant mutation associated chronic severe hepatitis B during lamivudine treatment.Methods Twenty-seven patients with lamivudine-resistant mutation associated chronic severe hepatitis B during lamivudine treatment were analyzed retrospectively.YMDD motif mutation was detected by gene chips or DNA sequencing.The pathological features of liver tissues from 8 patients undergoing liver transplantation were analyzed.The X2 test were used to perform the stafistical analysis.Results The YMDD motif mutations of 27 lamivudine-resistant patients were 5 cases of YVDD mutation,2 of YVDD+L180M,13 of YIDD mutation,4 of YIDD+L180M,1 of YVDD+YIDD mutations,2 of YVDD+YIDD+L180M,and there was no single L180 M mutation among patients.Twenty-seven patients were divided into cirrhotic group and noncirrhotic group according to whether they were diagnosed with cirrhosis before treatment.Compared to cirrhotic group,incidence of severe hepatitis was lower,prognosis was better,the age of patients was younger and hepatitis Be antigen(HBeAg)positive rate was higher in noncirrhotic group.There were two types of pathological features of liver tissues from 8 patients,which were active hepatic cirrhosis and massive or submassive hepatic necrosis with liver shrinking significantly.Conclusions Hepatic cirrhosis is a risk factor of lamivudine-resistant mutation associated chronic severe hepatitis B.There may be two mechanisms in lamivudine-resistant mutation associated chronic severe hepatitis B.

Objective To study the expression of extracellular matrix and polycystin-1 in ADPKD and their relation to cyst formation. Methods The expression of polycystin-1, fibronectin, laminin, type Ⅰ collagen, and type Ⅳ collagen were analysed in the normal kidney, fetal kidney and polycystic renal tissue by using immunohistochemical technique. Results The expression Of fibronectin, laminin, type Ⅰ collagen, and type Ⅳ collagen increased in polycystic renal tissue compared with normal kidney. The basement membrane lining cysts was markedly thickened. Type Ⅰ collagen was detected in the interstitium between cysts. Laminin, fibronectin and type Ⅳ collagen were localized in cyst basement membrane. The expression of polycystin-1 increased in polycystic renal tissue. The expression of extracellular matrix had significant correlation with the expression of polycystin-1. Conclusion The abnormal expressions of extracellular matrix and polycystin-1 exist in ADPKD. Abnormal expression of polycystin-1 may result in the alterations of extracellular matrix that is related to cyst formation.

Objective To analyze clinical courses and rescue therapies of adefovir-resistant chronic hepatitis B patients who had lamivudine resistance before and then changed to take adefovir dipivoxil. Methods 15 patients resistant to lamivudine were retrospectively analyzed, who had virological breakthrough after adefovir dipivoxil monotherapy and were treated with rescue therapy.Adefovir-resistant mutations were detected by direct sequencing of the HBV polymerase gene. Results 15 patients with former lamivudine resistance were treated with adefovir dipivoxil monotherapy for a median of 16 months, and 14 patients were found adefovir-resistant mutations at rtA181T/V and(or) rtN236T, only 1 patient was found multi-mutations at rtM204I + rtL180M + rtA181T. Rescue therapies were given to all the 15 patients after drug resistance. Among the 7 patients treated with lamivudine in combination with adefovir for 3 months,whose HBV DNA levels decreased (2.2±0.6)lg copy/mL on average, 5 patients achieved HBV DNA undetectable after 6 months combinative therapy. The HBV DNA levels of the 3 patients treated with entecavir decreased 2.8～3.5 lg copy/mL within 6 months treatment. Conclusion These preliminary data suggest the combination of lamivudine and adeforvir dipivoxil may be an effective rescue therapy for adefovir-resistant patients who have former lamivudine resistance.

Objective: To obtain polycystin-1 intracellular region. Methods: cDNA of polycystin-1 intracellular region was generated by PCR and then cloned into pProEX Hta, which was prokaryotic expression vector. After verified by sequencing, the recombinant was transformed into E.coli host to express and purify the fusion protein by affinity chromatography. Results: 660 bp cDNA of polycystin-1 intracellular region and 2.6×104 fusion protein were obtained. Conclusion: The fusion protein containing polycystin-1 intracellular region is obtained and is helpful for preparing anti-polycystin-1 monoclonal antibody.

Objective To analyze the clinical features and mutation patterns of HBV polymerase gene in patients with chronic hepatitis B(CHB) after the emergence of drug-resistance during Lamivudine(LAM) therapy.Methods LAM-resistant mutations were detected by direct sequencing of the HBV polymerase gene in hospitalized patients and outpatients of CHB with LAM-resistance in Changhai Hospital from Dec.2005 to Dec.2007.Clinical features after the emergence of LAM-resistant mutations were retrospectively analyzed.Results Two hundred and fifteen patients with CHB were diagnosed as LAM-resistant.Among them 192 patients were found to have LAM-resistant-associated mutations in the HBV polymerase gene.The mean value of serum HBV DNA was 6.25?1.31(log10copies/ml),the mean value of alanine aminotransferase(ALT) was 75U/L(ranged 19-821 U/L).ALT elevation and hepatitis recrudescence were found in 139 among 192(72.4%) patients.99.0%(190/192) patients had YMDD mutations.Four major mutation patterns of LAM-resistant HBV were identified as rtM204I(33.9%),rtL180M+rtM204V(26.0%),rtL180M+rtM204I(21.9%) and rtV173L+rtL180M+rtM204V(11.5%).The rtM204V mutation was accompanied more frequently by the rtL180M mutation compared with the rtM204I mutation(P0.05).Conclusions YMDD is the major mutation pattern of HBV polymerase gene after emergence of LAM-resistance.The mutation patterns of HBV polymerase gene are possibly not related to the clinical severity of CHB patients during LAM therapy.

Objective To evaluate the efficacy and safety of adefovir dipivoxil(ADV) monotherapy and ADV lamivudine(LAM) combined therapy for patients with LAM-resistant chronic hepatitis B.Methods 124 chronic hepatitis B patients with LAM-resistant mutations were enrolled in the present study.74 patients were treated with ADV combined with LAM therapy,and other 50 patients subjected to ADV monotherapy.There were no differences between the two groups in patients' baseline characteristics.Sequencing of the HBV polymerase gene was performed to determine LAM and ADV mutations occurred at baseline or during therapy.All patients were monitored with clinical examinations and routine laboratory tests during the therapy.Results The reduced logarithmic values of serum HBV DNA after 12-week and 24-week treatment were 1.99?0.64 and 2.61?0.80 in ADV group,obviously lower compared with those in ADV+LAM group(2.55?0.74 and 3.19?0.82,respectively,P

Objective: To prepare the monoclonal antibodies against polycystin 1 intracellular region and to study the distribution and expression of polycystin 1 in kidney tissues. Methods: Using the recombinant fusion protein containing polycystin 1 intracellular region as antigen, the hybrid cells secreting the monoclonal antibodies against polycystin 1 intracellular region were established by hybridoma technique. The distribution and expression of polycystin 1 in polycystic kidney, fetal kidney and adult kidney were investigated by immunohistochemical methods(standard EnVision method) with the monoclonal antibodies. Results: Four cell lines of hybrids steadily secreting the monoclonal antibodies against polycystin 1 intracellular region were established. The antibody titers were 1∶10 6. The 50th generation of these cell lines of hybrids still can secret the monoclonal antibodies and the titers remain similar. Polycystin 1 was weakly expressed in tubules and collecting ducts of normal kidney, the strong staining was seen at tubules of fetal kidney, and very strong staining of cyst lining epithelium of polycystic kidney was observed. Conclusion: The monoclonal antibodies against polycystin 1 intracellular region will be a useful tool in the studies of polycystin 1 structure and function.

Objective To discuss the influence of motor imagery therapy on the recovery of the stroke patients with aphasia.Methods Sixty cases of stroke patients with aphasia were divided into the treatment group and the control group according to the odd and even of last number.The single number was set into the control group.The double number was given to the treatment group.There were thirty patients in each group.Both groups received the conventional drug treatment and routine care of the Department of Neurology.On this basis,from the second day in hospital,the treatment group received the speech rehabili tation training with motor imagery therapy.Aphasia assessment was done for the two groups on the first day in hospital with CRRCAE.On the first day and after receiving ten-day treatment with motor imagery therapy,the grade of the severity for aphasia of Boston Diagnostic Aphasia Examination (BDAE) was done to assess the severity of aphasia in two groups.The scores of the two groups were compared.Results Before the treatment,there was no significant difference between the two groups.After receiving ten times treatment,both the two groups were improved.Compared with the control group,the treatment group became better.There was statistically significant difference between the two groups.Conclusions For the stroke patients with aphasia,motor imagery therapy has significant effect on their recovery.

Objective We reported the rehabilitative nursing points of patients with pectoralis minor muscle transplantation for recovery of thumb oppositional function. Methods We selected 10 patients who underwent pectoralis minor muscle transplantation for recovery of thumb oppositional function and gave them perfect preoperative examination and psychological nursing. We also implemented rigorous monitoring postoperation and auxiliary electric stimulation and functional exercises. The emphasis was put on the exercise of thumb oppositional function. Results The thumb oppositional function of all the ten patients recovered. The myodynamia reached level 4 and the shape of thenar was satisfying after 6 to 12 months of follow-up visit. Conclusions Sufficient preoperative psychological nursing and effective postoperative rehabilitative treatment had pivotal significance for the success of operation and the recovery of thumb oppositional function.

Objective To investigate the distribution of genotypes in chronic HBV infection (CHB) and acute HBV infection (AHB) patients in Shanghai. Methods Sixty-two patients with AHB and 73 patients with CHB admitted to ('hanghai Hospital of Shanghai between 2003 and 2007 were studied. Viral genotypes of all the patients were determined by direct gene sequencing.Meanwhile, epidemiological, clinical and biochemical parameters of all patients were collected. Mean values of different groups were compared by t test while frequency was compared by chi square test. Results The major prevalent genotypes in both AHB and CHB patients were genotype B and C (48.4% vs 51.6% in AHB patients and 26.0% vs 74.0% in CHB patients). The proportion of genotype B was higher in AHB patients compared to CHB patients (P= 0.02). Epidemiological factors and clinical outcomes were not statistically different among patients with different viral genotypes. The proportion of genotype C was much higher in CHB patients compared to AHB patients (P=0.006). The main transmission route of AHB was heterosexual interaction which was 18 out of 62 (29.0%), but in CHB patients, it was prenatal transmission which was 38 out of 73 (52.1%). Conclusions In shanghai, the main HBV genotypes in both AHB and CHB patients are genotype B and C. The proportion of genotype B is relatively high in AHB patients while proportion of genotype C is more common in CHB patients. There is no significant relationship between genotypes and the clinical outcomes of AI-IB patients.