Objective:To investigate the recent and forward effects of different operation patterns on recurrent calculus of intrahepatic duct.Methods:A analysis was made on previous history and operation patterns,distribution,effect and follow-up visit in 76 reoperation patients with recrudescent hepatolith between Jan,2000 and Feb,2008 in Affiliated Provincial Hospital of Anhui Medical University,retrospectively.The situation after operation was confirmed as the short-term curative effects and the follow-up results as long-term curative effects.Results:The average hospital stay was 10.4 days and the incidence rate of postoperative complications was 14.5%.The follow-up which lasted from 8 months to 8 years showed that the average recurernce rate was 38.2%.Besides,the rate of joint-type operation was lower than single-type operation (P

Objective To investigate the expression and significance of fragile histidine triad(FHIT) gene in the cholangiocarcinoma cell line QBC939.Methods The eukaryotic expression vector was transfected into QBC939,real-time RT-PCR was used to detect the change of FHIT expression,and direct counting method under microscope was used to detect the change in the biological behavior of cells.Results After transfection,the expression of FHIT mRNA increased by 2.53-fold,but the cell clones were significantly less [(11.2?1.3) vs.(10.5?1.1) vs.(6.3?1.0),P

Aim To investigate the expression of the lentiviral vector carrying a human apM1(hapM1) gene in 293 T cells and its bioactivity.Methods The CDS region of hapM1 gene was subcloned into the lentiviral vector from a cloning vector,and the transfer plasmid(PNL-hapM1-IRES2-EGFP)of the lentiviral vector was constructed.The three plasmids of the lentiviral vector: PNL-hapM1-IRES2-EGFP,HELPER,and VSVG were cotransfected to 293 T cells to produce the recombinant lentivirus;293 T cells were infected by the recombinant lentivirus and were identified with fluorescent microscope,RT-PCR and western blot.Inhibitory effect of the recombinant hapM1 on proliferation of vascular smooth muscle cells(VSMCs) was identified.Results The sequencing results showed that the sequence of the subcloned hapM1 gene was completely identical to that reported in GeneBank.The virus titre of the recombinant hapM1 lentivirus was 2.0?105 TU?L-1.After the recombinant lentivirus infected 293 T cells,RT-PCR and Western blot results showed that the expression of hapM1 in the hapM1-293 T group could be measured.The recombinant hapM1 inhibited the proliferation of VSMCs.Conclusions The recombinant lentivirus carrying a human apM1 gene was constructed successfully.The recombinant hapM1 could be expressed in 293 T cells and had its bioactivity.

BACKGROUND: AdiponecUn (apM1) has been a new target for gene therapy of ischemic diseases; apM1 gene cloning is the key of successful apM1 gene therapy. There are two main gene cloning methods: directional cloning and T-A cloning method. Directional cloning method is complicated, while the T-A cloning method is relatively simpler, with higher successful rate. OBJECTIVE: T-A cloning of apM1 gene coding region was applied to verify the sequence in comparison with GenBank. DESIGN, TIME AND SETTING: The gene verification experiment was performed at the laboratories of Fujian Hypertension Research Institute which is in The First Affiliated Hospital of Fujian Medical University and Academy of Integrative Medicine which is in Fujian University of Traditional Chinese Medicine, from June 2006 to December 2008. MATERIALS: Adipose tissue sample of omental fat pad was obtained from a surgical patient in the First Affiliated Hospital of Fujian Medical University. Trizol produced by Invitrogen company; M-,-MLV, Gel Extract Kit produced by PROMEGA company; Taq enzyme produced by TIANGEN company; Restriction Endonucleases BamH Ⅰ and Sal Ⅰ, pMD18-T Vector produced by TAKARAcompany were used in this study.METHODS: Total mRNA was extracted from human greater omentum adipose tissue. The coding region of human apM1 (hapM1) gene was amplified by RT-PCR. The coding region of hapM1 gene was cloned into pMD18-T vector .The recombinant plasmid was identified with restriction enzyme digestion analysis and nucleotide sequencing. MAIN OUTCOME MEASURES: The electrophorasis verification of the recombinant plasmids coding target gene using double enzyme digestion, comparison of the similarity between the sequences of the plasmids and hapM1 in GenBank. RESULTS: Sense and antisense coding region of hapM1 gene were cloned. The sequencing results showed that the sequences of the cloned DNA were completely identical to that of hapM1 in GenBank.CONCLUSION: The coding region of hapM1 gene was successfully cloned.

Marfan's syndrome(MFS) is a relatively common autosomal dominant hereditary disorder with prominent manifestations in the skeletal, ocular, and cardiovascular systems. MFS has a high penetrance. Recent research on the pathogenesis and diagnosis and therapy of MFS has made a lot of progress. So we reviewed the advance on the relation of molecular genetics and phenotype of MFS.We discussed the molecular hasis, gene mutation and location, diagnosis, clinical features and therapy. Now the detection on the molecular level has been a very important method in diagnose MFS very early or before bom.

[Objective] To observe the effect of serum containing Bushen prescription (BP) on inhibition of chondrocyte proliferation and nitric oxide (NO) production in chondrocyte induced by interleukin-1 (IL-1). [Methods] Four New Zealand rabbits were administered BP 2.5mL (1.15kg/L) for 3 days for the preparation of BP-containing serum. Rabbit chondrocytes were cultured in-vitro and were added with DMEM culture containing IL-1 1, 10 and 20 ?g/L to examined the inhibition concentration of IL-1 for chondrocyte proliferation. After then, BP group was added with 20 ?g/L IL-1 and various concentrations of BP-containing serum and the model group with 20 ?g/L IL-1 and various concentrations of blank serum. Alter 3-day culture, chondrocyte proliferation were dectected by thiazolyl blue colorimetry (MTT) and NO content by Griess method. [Results] The inhibition concentration of IL-1 for chondrocyte proliferation was 20 ?g/L; various concentrations of BP-containing serum could counteract chondrocyte proliferation and inhibit NO pnxluction in chondrocyte induced by IL-1. [Conclusion] BP can counteract chondrocyte proliferation directly or indirectly by the mediation of NO, which may be one of its therapeutic mechanisms in treating osteoarthritis.

Objective To explore the value of detecting serum cytokeratin 19 fragmen 21-1 (CYFRA21-1) and squamous cell carcinoma antigen (SCCAg) in the diagnosis of patients with nasopharyngeal carcinoma.Methods 79 patients with nasopharyngeal carcinoma from October 2013 to October 2015 as case group and 145 patients with nasopharyngeal chronic inflammation as control group were collected.The CYFRA21-1 level was detected by the electrochemical luminescence immunoassay and the SCCAg level was detected by the enzymoimmunoassay.Comparisons of diagnostic value of CYFRA21-1 and SCCAg to nasopharyngeal carcinoma were conducted according to pathological diagnosis.Results The serum CYFRA21-1 and SCCAg levels in case group were (12.37±1.65) g/L and (1.65±0.37) ng/ml,and the control group were (4.26±1.99) g/L and (1.22±0.45) ng/ml,with statistically significant differences (t =8.093,P =0.002;t =4.287,P =0.043).The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of CYFRA21-1 combined with SCCAg in the diagnosis of nasopharyngeal carcinoma were higher than those of CYFRA21-1 and SCCAg respectively,and the difference all were statistically significant (all P ＜ 0.05).Conclusion Detecting serum CYFRA21-1 combined with SCCAg in the diagnosis of nasopharyngeal carcinoma has high sensitivity and good specificity,which can significantly improve the diagnostic accuracy.

Objective To explore the correlation of traditional Chinese medicine (TCM) syndrome types of hypertension with carotid artery intima-media thickness (IMT) and plasma lipoprotein-associated phospholipase A2 (Lp-PLA2),so as to provide a TCM therapy for early prevention and treatment of hypertension and its complications in clinic.Methods We collected a total of 170 cases of hypertension patients,55 with kidney-essence insufficiency syndrome,50 with ascendant hyperactivity of liver yang syndrome,and 65 with phlegm-damp retention in middle energizer syndrome.IMT was measured by the carotid Doppler ultrasound,and plasma Lp-PLA2 expression was examined by enzyme-linked immunosorbent assay.And then the differences of IMT and Lp-PLA2 expression among the three groups were compared and analyzed.Results There were significant differences of carotid artery IMT and Lp-PLA2 among the three groups(P＜ 0.01),and the number of patients with carotid atherosclerosis and the expression level of Lp-PLA2 in the group of phlegm-damp retention in middle energizer syndrome were higher than those intheothertwogroups(P＜0.01).Conclusion Differentiation treatment based on phlegm may be more conducive to the prevention and treatment of hypertension and its complications.

Objective To study the effects of ulinastatin (UTI) on the inflammatory cytokines in the blood serum of rats after severe burn. Methods A total of 96 SD rats, inflicted with 30% TBSA full thickness skin burn, were randomly divided into simple burn group (group B) and UTI treated group (group UTI). The serum contents of IL 1?, IL 6, and TNF ? were determined at 0, 1, 3, 6, 12, and 24 h before and after burn injury. Results In group B, the levels of serum IL 1?, IL 6, and TNF ? were significantly higher at 3 h and peaked at 6 and 12 h after burn. In group UTI, the levels of serum IL 1?, IL 6, and TNF ? were significantly lower than those in group B ( P