Objective To construct an infectious full-length cDNA clone of enterovirus 71(EV71)and develop a technological platform for study on vaccine development as well as molecular virology of EV71.Methods According to the nucleotide sequence of EV71 strain 085 isolated in China,four pairs of primers were designed for amplification of four end to end overlapping subgenomic cDNA fragments,the cDNA fragments were directional cloned into pBluescript SK(+)vector,and the virus genome cDNA clone was obtained by ligation orderly.The rescued virus of parental strain 085 from RNA transfected host cells was identified by RT-PCR,IFA,titration as well as transmission electron microscope(TEM)after the transcription of the full-length cDNA clone in vitro.Results The full-length cDNA clone was constructed successfully,and the typical CPE was observed after its transcription into Vero cells.The rescued virus with 20-30 nm in diameter can not only be neutralized by EV71 special anti-serum but also react with anti-EV71 monoclonal antibody that virus infected cells stained with FITC can be detected by IFA.After amplification from the total RNA extraction of virus infected cells by RT-PCR with EV71 special primers,the 226 bp products can be detected.The growth curve showed that the rescued virus can propagate in Vero cells stably with a titer of 4.5 ～6.0 lgCCID50/ml during 8 passages.The plaque formed by rescued virus is identical as parental virus in morphology but smaller in size.Conclusion An infectious full-length clone of EV71 was developed successfully,which will be used for further study on pathogenesis and vaccine development of EV71.

0.9 mg/L in 6 patients.Kayser-Fleischer ring were found in 85.5 % of all the patients. The abnormal hepatic function in the liver type HLD was more common than that of in the brain type. The liver injury was detectable by B mote ultrasonic wave in different type HLD.MRI examination was taken in 79 patients, 65 of them had showed the symmetry abnormal signs in basal ganglia. Conclusions CP has independent diagnostic values when its content ≤0.08 g/L. Kayser-Fleischer ring is an excellent discriminatory test for the diagnosis of HLD patients with neurological or psychiatric symptom. 24 h urine copper is the best single screening test because it increases over 100 ?g/24 h in all patients who were taken the test. The B mote ultrasonic wave test for the liver and MRI for brain were helpful in detecting the damage in the liver and brain.

Objective To compare the therapeutic result of endoscopic po lypectomy with colectomy for polypoid malignant colorectal tumor. Methods A tot al of 61 cases of polypoid malignant colorectal tumor including thirty－ nine pa tients treated by endoscopic polypectomy and surveillance,twenty－ two patients treated by colectomy.Follow－ up ranged from 28 to 168(mean 64) months. Results During the follow－ up period,one patient died relating to this disease,two rec urred at the original site,and one metastasized to the liver.The distribution of the adverse outcome had no significant statistical difference between the group of endoscopic polypectomy and colectomy;significant difference existed between the sessile and pedunculated malignant polyps (P=0.006),as well as between favou rable(carcinoma invading the muscularis mucosa or above,limited to the head of t he pedunculated polyp,polypectomy margin negative,and well and moderately differ entiated)and unfavourable(without those criteria) histologic characteristics(P=0 .002).Conclusion The polypoid malignant colorectal tumors with favourable his tologic characteristics could be treated by endoscopic polypectomy alone,tumors with unfavourable histologic characteristics should be treated by colectomy.

BACKGROUND: It is of great importance to study the genotype distribution of hereditary ataxia in understanding its epidemiologic rule and pathogenetic pathway.OBJECTIVE: To analyze the distribution of different genotype of hereditary ataxia in south China.DESIGN: A case-control observation.SETTING: Department of Neurology, the First Affiliated Hospital of Sun Yat-sen University.PARTICIPANTS: Forty-three patients (26 males and 17 females) with hereditary ataxia from 36 families and 38 patients with sporadic hereditary ataxia (24 males and 14 females) were selected from the Outpatient Clinic of Neurogenetics, Department of Neurology, the First Affiliated Hospital of Sun Yat-sen University between September 1998 and September 2002. At the same time, 60 healthy individuals from the patients' families and 44randomly-selected healthy physical examinees were taken as controls. All the participants were enrolled voluntarily.METHODS: The fragments of trinucleotide repeats at different sites of mutant genes were amplified with polymerase chain reaction (PCR), and then the lengths were calculated with polyacrylamide gel electrophoresis and imaging analytical software. The repeated numbers of trinucleotide repeats in all the normal and abnormal amplified alleles were calculated respectively.MAIN OUTCOME MEASURES: Different genotype distribution in patients with hereditary ataxia.RESULTS: All the subjects were involved in the analysis of results. Of the detected patients with hereditary ataxia, the Machado-Joseph disease/spinocerebellar ataxia (SCA) 3 was the most common type of autosomal dominant SCA in South China, which was 42.0%, and was followed by SCA2 (7.4%), SCA1 (4.9%), SCA7 (3.7%), SCA6 (2.5%), SCA12 (1.2%).No patient was detected to have SCA8 SCA 10, SCA 17 dentatorubropallidoluysian atrophy (DRPLA) and Friedreich ataxia (FRDA).CONCLUSION: Autosomal dominant SCA3 is the most familiar genotype in South China. Clinical detection of hereditary ataxia should be done firstly aiming at the SCA3 genotype.

Analogy is to construct the theory of traditional Chinese medicine in clinical thinking method,which the proper use of this thinking is helpful to improve the clinical thinking of Chinese medicine,contributes to the development of disease differentiation and treatment ideas.This article tried to analyze the diagnosis and treatment of kidney diseases with Professor Zhu Zongyuan on the process,to explore the analogy on disease differentiation treatment guide.

Objective To investigate the expression of forkhead box protein P1(FOXP1)in gastric mucosa-associated lymphoid tissue(MALT)lymphomas and its relationship with histological morphology and prognosis. Methods In this study.samples of 43 MALT lymphoma were studied histologically and divided into monomorphic histology group and polymorphic histology group according to their cellular features.The expressions of FOXP1 and NF-κB in gastric MALT lymphoma were evaluated immunohistochemically by two-step method of Envision,and the clinicopathological features and prognosis were analyzed retrospectively.Results The nuclear expressions of FOXP1 in 43 cases with gastric MALT lymphoma were 44%(19 of 43),including strong immunoreactivity in 7 cases and moderate immunoreactivity in 12 cases.There were 4 cases with positive immunoreactivity in moninorphic histology group and 15 cases in polymorphie histology group,and the difference was statistically significant(15%vs.88%,P＜0.01).All the postoperative recurrent cases were strongly positive with FOXP1 stain,and it was closely with FOXP1 expression(P＜0.01).The median survival time(26 months)in polymorphic histology group was significantly shorter than that(123 months)in monmorphic histology group(P＜0.01),and the median survival time was significantly longer in negative FOXP1 expression group than that in moderate FOXP1 expression group and in strong FOXP1 expression group(115 vs.55 vs.12 months)(P＜0.05).similarly,the median survival time in nuclear factor kappa B(NF-κB)expression group was significantly shorter than that in negative NF-κB expression group(26 vs.131 months)(P＜0.01).The median survival time in stageⅠ(98 months)and stage Ⅱ(121 months)was significantly longer than that in stage Ⅱ E+Ⅳ(33 months)(P＜0.01).By multivariate COX regression analysis.FOXP1 nuclear expression and clinical stage were independently prognostic factom. Conclusion FOXP1 expression may be used as a biomarker for the assessment of malignant transformation to diffuse large B-cell lymphoma(DLBCL)and predicting prognosis.

Objective To construct a recombinant adenovirus contained respiratory syncytial virus F gene fragment. Methods The F gene fragment was amplified by RT-PCR. Then the recombinant shuttle plasmid pShuttle-CMV/F was constructed. The iinearized pShuttle-CMV/F was transformed into BJ5183-AD-1 electroporation competent cells which contained plasmid pAdEasy-1. The obtained homologous recom-binant plasmid was named pAdEasy/F, and transfected into 293 cells. Then the recombinant adenovirus rAd/F was obtained, and it was tested by electron microscope, RT-PCR, Western blot and immuno-fluores-cence assay(IFA). The vires titer and genetic stability were also studied initially. Results The recombi-nant adenovirus rAd/F was constructed, which was visualized as typical adenovirus morphology under elec-tron microscope. The transcription and expression of RSV F gene fragment integrated in the rAd/F were con-finned by RT-PCR, Western blot and IFA. Conclusion The recombinant adenovirus rAd/F contained RSV F gene fragment is successfully constructed and lay an important foundation for the further vaccine study.

Objective To study the molecular genetic diagnosis and clinical characteristics of spinocerebellar ataxia type 6 (SCA6).Methods 43 patients with autosomal dominant SCA from 36 families and 38 sporadic SCA patients were enrolled in the study. SCA6 (CAG)n dynamic mutations were detected by polymerase chain reaction (PCR). Abnormal allele fragments were sequenced and repeated numbers were calculated. The clinical data of two cases with SCA6 were analyzed.Results CAG repeat of normal SCA6 allele ranged from 10 to 13. CAG repeat of abnormal SCA6 allele expanded to 25 in one familial patient and 24 in one sporadic patient in our study. The basic characteristics of these SCA6 patients were slowly progressive cerebellar ataxia, nystagmus and dysarthria.Conclusion Diagnosis of SCA6 can be confirmed by detection of abnormal CAG repeat expansion. There is no obvious difference of clinical features between SCA6 and other SCA subtypes.

Objective To study the feasibility of re-operation of gynecological laparoscopy.Methods Clinical records of 47 patients with gynecological diseases undergoing a re-laparoscopy from December 1995 December 2003 were analyzed retrospectively.Results The re-operations of laparoscopy were successfully accomplished in 46 patients(97.9%) while a conversion to open surgery was required in 1 patient.The re-operation time was 30~80 min(mean,50 min),and the in-hospital stay was 2.5~5 days(mean,3.5 days).No long-term complications were noted in 44 patients on a postoperative follow-up for(1~4) years.Out of 19 patients with infertility, 1 patient had a natural pregnancy and 6 patients were pregnant with the help of artificial reproductive techniques.Supportive medical treatment had been given in patients with endometriosis postoperatively,3 of whom experienced symptomatic improvements and 1 of whom had a natural pregnancy.Conclusions For patients with a history of laparoscopy,a re-operation of gynecological laparoscopy is safe and feasible.