OBJECTIVETo investigate the apoptosis regulation on hepatoma cells by HBx between genotype B and C.

METHODSGenotype B and C HBx gene fragments were amplified and inserted into green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP-XB and pGFP-XC. The pEGFP-C1, pGFP-XB and pGFP-XC were introduced into Bel-7402 cells by Fugene HD to obtain Bel-7402 cells expressing GFP. The transcription and expression of HBx gene were demonstrated by RT-PCR and Western Blot analysis. Bel-7402, Bel-7402/GFP, Bel-7402/GFP-XB, Bel-7402/GFP-XC cells were treated with adriamycin (2.5 microg/ml), and the apoptosis of the cells was determined by trypan blue exclusion, and flow cytometry analysis.

RESULTSRT-PCR and Western Blot analysis showed that HBx genes of genotypes B and C were transcribed and expressed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in Bel-7402, Bel-7402/GFP cells while no significant cell death was observed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Flow cytometry analysis indicated that no significant differences of apoptosis rates of Bel-7402/GFP-XB (3.87%) and of Bel7402/GFP-XC (4.01%) were observed (P > 0.05), moreover, no significant differences of Bel-7402/ GFP-XB (3.87%), Be17402/GFP-XC (4.01%) and of the untreated cells. Apoptosis rates in Bel-7402/GFP-XB (3.87%), Bel-7402/GFP-XC (4.01%) cells were significantly lower than those in Bel-7402 (27.05%) and Bel-7402/GFP (29.14%) cells at 48 hours after the adriamycin treatment (P < 0.01).

CONCLUSIONSBel-7402 cell lines expressing GFP, GFP-XB and GFP-XC fusion proteins were successfully established. HBV X protein blocks adriamycin-induced apoptosis of Bel-7402 cells. There is no difference between HBx of genotype B and C in inhibiting apoptosis induced by adriamycin.

Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Hepatitis B ; drug therapy ; physiopathology ; virology ; Hepatitis B virus ; classification ; drug effects ; genetics ; metabolism ; Humans ; Trans-Activators ; genetics ; metabolism

Objective To evaluate the clinical role of F-18 fluorodeoxyglucose (FDG)-positron emission tomography (PET) for the diagnosis and staging of prostate cancer.Methods 18F-FDG PET/CT and 99Tcm-MDP ECT whole body bone scan were both performed in 40 patients with prostate cancer proved by biopsy or surgery.We then evaluated the diagnostic value of 18F-FDG PET/CT for prostate cancer.Results Of 40 cases,17 cases were detected by 18 F-FDG PET/CT with a sensitivity of 42.5％.Of the 17 cases with lymphoma metastasis,8 cases were detected by CT while 15 cases were detected by 18F-FDG PET/CT.The sensitivity of 18F-FDG PET/CT is 88.2％.Though 18F-FDG PET/CT and 99Tcm-MDP bone scan have a similar sensitivity in detecting bone metastases,18 F-FDG PET/CT has better specificity and accuracy in detecting bone metastases than that of an 99Tcm-MDP bone scan.Conclusions Though the diagnostic value of 18F-FDG PET/CT in detecting primary focus of prostate is limited,it still has an important role in detecting lymph node metastases and bone metastases.18F-FDG PET/CT has its superiority in prostate cancer staging.It can also help clinicians to select the regimen of treatment.

AMP-Activated Protein Kinases ; genetics ; Heart Diseases ; Humans ; Syndrome

Objective To assess the effect of pharmacological and non pharmacological interventions on Chinese subjects with impaired glucose tolerance (IGT). Methods In this 3 year prospective study, 321 subjects aged above 25 years with IGT were included. IGT was defined by 1985 WHO criteria utilizing a 75 g oral glucose tolerance test (OGTT). The subjects were divided into control (C), diet plus exercise (D+E), Acarbose (Glucobay, A) and Metformin (M) groups. The subjects in the group D+E underwent an individually designed diet and exercise program, the importance of which was reiterated annually. The group C only received conventional education on diabetes prevention. The two pharmacological groups were orally given Acarbose (50 mg, t.i.d) and Metformin (0.25 g, t.i.d) respectively. OGTT, weight, height, blood pressure, lipids were measured yearly during the follow up. The t test, Chi square test and proportional hazard regression analysis were used to analyze the risk reduction in diabetes conversion from each group. Results The baseline data of 4 groups had no statistical differences. By the end of study, both the fasting plasma glucose (FPG) and the 2h postprandial plasma glucose (2hPG) in group C elevated (FPG from 6.02 mmol/L to 6.59 mmol/L, 2hPG from 8.83 mmol/L to 9.13 mmol/L), and the annual diabetes incidence was 11.6%. The corresponding changes in group D+E were FPG from 6.11 mmol/L to 6.21 mmol/L, PG2h from 9.28 mmol/L to 8.98 mmol/L, and 8.2% of annual diabetes incidence. In contrast, both the FPGandthe2hPG significantly decreased in group A (FPG from 6.03 mmol/L to 5.47 mmol/L, 2hPG from 8.34 mmol/L to 7.21 mmol/L)andingroup M (FPG from 6.01 mmol/L to 5.85 mmol/L, 2hPG from 9.05 mmol/L to 7.92 mmol/L). Theannualdiabetesincidencedecreasedto2.0% in group A, and 4.1% in group M. Proportional hazard regression analysis showd that the annual diabetes incidence was positively correlated with the baseline 2hPG and body mass index (OR=1.33, P=0.006 and OR=1.11, P=0.008, respectively), and negatively correlated with group C and group M (OR=0.12, P=0.0001 and OR=0.23, P=0.0002, respectively). Conclusion Thenaturaldiabetesincidenceis 11.6% in IGT population, and 8.2% in population with conventional diet and exercise interventions, between them there is no significant difference. The pharmacological interventions with Acarbose or Metformin significantly decrease diabetes incidence of IGT population.

Basic fibroblast growth factor(bFGF)is a polypeptide cell growth factor,which has exten- sive physiological functions.With the in-depth study,the exploration of relationship between basic fibroblast growth factor and tumor has been developing rapidly in recent years,It shows a good prospect in diagnosis and treatment of tumor.There is a brief overview about the molecular biology of bFGF,its expression in tumor and the relationship between bFGF and tumorigenesis.

Objective The aim of this study was to measure quantitatively the TSP-4 mRNA expression and its significance in gastric carcinoma was evaluated by correlating its expression with clinicopathological features, microvessel density(MVD) and MMP-9.Methods Eighty-two patients with gastric carcinoma were recruited in this study. TSP-4 mRNA expression in gastric carcinoma and adjacent tissue was detected by real-time PCR. CD34 and MMP-9 protein expression were examined by immunohistochemistry. MVD was determined according to CD34-positive tubular structures. Results The expression level of TSP-4mRNA in gastric carcinoma was obviously higher than those of adjacent tissue(P=0.03) and it was associated with tumor size, histological type, lymph node metastasis, MVD and MMP-9(P=0.002, P=0.031, P=0.014,r＝0.67 P<0.01, P=0.008),but not sex, age and depth of invasion. (P=0.53,P=0.57,P=0.15).Conclusion TSP-4 may play an important role in occurrence and progression of gastric carcinoma and that the effect of TSP-4 on tumor growth and metastasis may be exerted through regulation of angiogenesis and MMP-9 expression in gastric carcinoma.

OBJECTIVETo compare the clinical efficacy on the motor impairment of knee joint after tracture operation between the combined therapeutic method of relaxing needling manipulation and exercise therapy and the simple exercise therapy.

METHODSSixty-four patients after the operation for the fracture of femoral shaft were randomized into a relaxing needling combined with exercise therapy group (group A) and an exercise therapy group (group B), 32 cases in each one. In the group A, the relaxing needling manipulation was applied to the local painful area of knee or the stiff soft tissues. Additionally, the exercise therapy was used in combination. In the group B, the exercise therapy was applied simply. Hospital for special surgery (HSS) pain score, the range of movement (ROM) of knee joint and Lysholm score were compared before and 60 days after treatment in the patients of the two groups. The efficacy was compared between the two groups.

RESULTSAfter treatment, HSS pain score, ROM and Lysholm score were all improved in the two groups, presenting the significant differences as compared with those before treatment (all P<0. 05). The results in the group A were better than those in the group B (all P<0. 05). The total effective rate was 96. 9% (31/32) in the group A, which was better than 75. 0% (24/32) in the group B (P<0. 05).

CONCLUSIONThe combined therapeutic method of relaxing needling manipulation and exercise therapy achieves the significant efficacy on the motor impairment of knee joint after the operation for the fracture of femoral shaft, superior to the simple exercise therapy.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Combined Modality Therapy ; Exercise Therapy ; Female ; Fractures, Bone ; physiopathology ; surgery ; therapy ; Humans ; Knee Joint ; physiopathology ; Male ; Middle Aged ; Treatment Outcome

Objective To express and purify recombinant Tp0136 epitope fragment, and study the immunity activity. Methods The Tp0136 selective fragment(Tp0136B) gene was devised by the surface property analysis, solvent-accessible suface calculateions, secondary structure function region analysis, and was inserted between the sites of Nde Ⅰ and Not Ⅰ in pET22b ( + ) . The recombinant plasmid was expressed in E. coli BI21. After nickel ion metal affinity chromatography, the antigenic and immune reactivity of rTp0136B was confirmed. Then indirect ELISA with the rTp0136B as coating antigen was performed to detect the anti-Tp0136 antibody in sera from 100 normal human controls and 131 primary syphilis patients. Results The rTp0136B was soluble expressed with a molecular weight of about 28 000 and was obtained with a purity of ＞98% by chromatography. Western blot proved that the rTp0136B could specifically react with anti-Tp0136 polyclonal antibody. Specific humoral response was elicited by the recombinant protein in Japan negative. The positive detection rate in sera from primary syphilis patients was 85.5%. Conclusion This result suggested that the recombinant Tp0136 epitope fragments have a satisfactory immunocompetence,which may have applications in the serodiagnosis of primary syphilis.

BACKGROUND: Cervical spinal cord injury without radiographic abnormality (SCIWORA) is a special type of spinal cord injury. Because of the particularity of children in the period of growth and development on the aspects of anatomy and biomechanics, their injured mechanism and therapeutic method differ from those of adults.OBJECTIVE: To explore clinical feature and therapy of first-episode cervical SCIWORA in children.DESIGN: Retrospective analysis and self pre-and post-control observation.SETTING: Department of Spine Surgery, Second Affiliated Hospital,Harbin Medical University.PARTICIPANTS: Totally 14 patients with cervical SCIWORA, who were treated at the Departmentof Spine Surgery, Second Affiliated Hospital,Harbin Medical University between June 1997 and June 2003. Inclusive criteria: ①patients who had dysfunction of sensation, exercise and sphincter to different degree in clinic, ②patients who had no fracture dislocation after cervical vertebra X-ray plain film and MRI examination. The MRI examination showed the sign of spinal cord injury. T1WI spinal cord became thick. T2WI distributed at strip-shape high-signal region along spinal prosenchyma and spread upward and downward. There were 3 patients with complete spinal cord injury and 11 patients with incomplete spinal cord injury, among the patients with incomplete injury, there were 6 patients with central cord syndrome (CCS), 3 with Brown-Sequard syndrome and 2withfrontal spinal cord injury syndrome.METHODS: Retrospective analysis was performed in 14 children patients with cervical SCIWORA. Thirteen patients were not treated with operation and one patient was treated with cervical posterior atlantoaxial fusion. MAIN OUTCOME MEASURES: ASIA grading before and after treatment in patients.RESULTS: There were 1 dead patient and 13 cases with 3-year follow up averagely. Three patients were with insignificant amelioration, and others had recovery of limb function to different degree. According to ASIA grading standard, before treatment there were 3 cases in A grade, 5 cases in B grade, 5 cases in C grade and 1 case in D grade, and after treatment there were 1 case recovering to B grade, 2 cases to C grade, 5 cases to D grade and 2 cases to E grade.CONCLUSION: Restoration of neurofunction of children with cervical SCIWORA has closely correlation with degree of primary injury of spinal cord. Most of the patients were not treated with operation, and those with obviously instable cervical vertebra can be treated with operation.

Folium Perillae (FP) is a traditional Chinese materia medica, which has been used for treating inflammatory diseases. In order to clarify the material basis of FP' s pharmacological activity and anti-inflammatory mechanism, we presented a review about some of the primary chemical components in FP, such as volatile oils, flavonoids, anthocyanins, phenolic acids, glycosides, triterpenes and steroids, and about the anti-inflammatory activity of FP extract and its major mechanism, such as regulating the viability and function of innate immune cells, controlling the balance of helper T cells, based on related research in recent years. This study aims to provide reference for further research and development of new drugs based on FP.