Purpose:To investigate the antitumor effect of 5-fluorocytosine/cytosine deaminase (5FC/CD) suicide gene system combined with heat shock protein 70 peptide complexes(HSP70-PC) tumor vaccine.Methods:5-FC was administrated intraperitoneally after recombinant adenoviruses carrying E.coli CD gene were injected into murine tumor tissue. At the same time,mice bearing tumor were inoculated s.c. with HSP70-PC. Results:The combined therapy led to reduction in tumor growth, and complete tumor regression was observed in 70% of the mice. After the combined therapy, the tumor tissue showed more CD + 4 and CD + 8T cells infiltration, and the murine CTL activities increased significantly. Conclusions:The therapy combining 5-FC/CD system with HSP-PC tumor vaccine is very effective in murine gastric cancer, and has potential for clinical usage.

Objective To discuss the effect of hyperthermia on tight junctions of the endothelial cells in the blood-brain barrier and explore the molecular mechanism. Methods An in vitro blood-brain barrier model was established by coculture of ECV304 and astrocytes. Transendothelial resistance (TER) of in vitro blood-brain barrier was determined by Millicell-ERS system. The morphological change of tight junctions of the endothelial cells in the in vitro blood-brain barrier was determined by the method of silver staining. The expression levels of zonula occluden 1 (ZO-1) and occludin were analyzed by means of semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western bloting. Results After two hours at 43℃, the mean value of TER was decreased from (321.30? 58.59) ??cm2 to (65.67?6.02) ??cm2. The integrity of tight junctions was destroyed and the expressions of ZO-1 and occludin decreased significantly. Conclusions Hyperthermia can destroy the tight junctions of the endothelial cells in the in vitro blood-brain barrier. The expression decrease of ZO-1 and occludin induced by hyperthermia is one of the most important molecular mechanisms.

Objective To investigate the effects of tea polyphenol on oligodendrocyte(OLG) in the cerebral ischemic rats. Methods Male Sprague Dawley rats were randomly divided into sham-operated group, control group and tea polyphenol group. The models of whole-brain ischemia were established . Then the rats of tea polyphenol group were divided another 3 groups and injected intraperitoneally with tea polyphenol at diffenrent dose (25 mg/kg, 50 mg/kg, 100 mg/kg, respectively) once a day for seven days . The expressions of precursor OLG cells, pedo-OLG cells and mature OLG cells in the cortex were examined by immunohistochemistry and the positive cells were counted.Results Compared with sham-operated group, the number of precursor OLG cells increased and the number of pedo-OLG cells and mature OLG cells decreased significantly in the control group (all P

Objective To explore the changes and influencing factors of serum NT-proBNP level in aged patients with severe brain dysfunction in acute phase, and the corresponding clinical significance. Methods The serum NT-proBNP and cTn-I levels of patients with severe brain dysfunction at day 1, 3, 5 and 7 were measured respectively. The correlation between acute neurological dysfunction caused by brain dysfunction and serum NT-proBNP and cTn-I levels were analyzed, and the impact on prognosis was explored. Results The serum NT-proBNP levels were significantly higher in death group at day 3[(759±341)ng/L], 5[(1980±839)ng/L] and 7[(2490±1862)ng/L] than in survival group[(594±612)ng/L,(733±424)ng/L,(315±346)ng/L]. Serum NT-proBNP and cTnI levels were associated with progressive cerebral edema in both groups. Location of early intracranial lesions was significantly different between two groups. Death group had higher ratio of intracranial lesions in basal ganglia and brainstem than did survival group. High serum NT-proBNP level after day 7 suggested poor prognosis. Serum NT-proBNP level was not associated with serum cTn-I level. Conclusions Progressively increased serum NT-proBNP level in aged patients with severe brain dysfunction in acute phase suggests poor prognosis. The increased degree of serum NT-proBNP in aged patients with severe brain dysfunction in acute phase is associated with the location of intracranial lesions.

Objective To investigate the causes and managements of capsular perforation in transurethral resection of prostate(TURP) for benign prostatic hyperplasia (BPH). Methods Clinical data of 893 cases who received TURP from February 2002 to February 2011 were analyzed retrospectively.Results In 893 cases, capsular perforation in 16 cases, among them, tissue covered perforation in 10 cases, dissociative perforation in 3 cases, perforation under bladder trignne in 3 cases. Six cases occurred with obviously extravasation, 10 cases with transurethral resection syndrome (TURS) or omen of TURS. Two cases were ceased operation and received TURP again after 2 weeks, 2 cases transferred to open surgery suprapubic prostatectomy, 12 cases kept up operation with lower bladder douche pressure. Two cases with postoperatire bleeding underwent open surgery hemostasis cured. All cases were discharged from hospital after recovery. Conclusions The type and extent of capsule perforation must be judged correctly,corresponding treatment must be given, prevention and treatment of TURS must be taken notice of, if capsule perforation occurs when TURP operation in progress. Less haematoma, clearview and normal anatomic structures can decrease the incidence of perforation.

AIM: To establish a HPLC method for the determination of tanshinone ⅡA in Compound Danguifukang Capsule. METHODS: Analytic column: Reliasil C 18 (5 ?m, 250 mm?4.6 mm), protective column:(5 ?m, 12.50 mm?4.6 mm); mobile phase: methnaol-ultrapure water (80∶20); detective wavelength was at 270 nm; temperature was at 30 ?C ; the flow rate was about 1.0 mL?min -1 . RESULTS: The standard curve showed a good linearity within the range of 1?10 -2 -0.16 ?g, r=0.999 5. The average recovery was 99.73% and RSD was 2.91% . CONCLUSION: The method is feasible, and simple to operate and suitable for the quality control of Compound Dangguifukang Capsule.

Objective To observe and compare the clinical effect of traditional Chinese medicine (TCM)combined with transurethral resection of prostate(TURP) and simple TURP in treating benign prostatic hyperplasia (BPH). Methods 60 patients with BPH were randomly divided into two groups. The treatment group(n=30) was given the TCM combined TURP and the control group(n = 30) was given TURP alone. The effects were observed.Results Improvement of lower urinary tract symptoms(LUTS), the international prostate symptom score (I-PSS) of the treatment group was better than that of the control group with significant difference (P ＜ 0.01). The time and quantity of bladder irrigation, incidence of bladder spasm, incidence of constipation after operation of the treatment group were less than that of the control group with significant difference(all P ＜0.05). Catheter retention time,postoperative bleeding, urinary retention were less and had no significant difference between the two groups (all P ＞0. 05). Conclusion The effect of TCM combined with TURP was significantly superior to that of simple TURP; and adverse reaction after treatment was less than that in the control group.

AIM: To establish a HPLC method for the determination of tanshinone Ⅱ A in Compound Danguifukang Capsule. METHODS: Analytic column: Reliasil C18(5μm, 250mm×4.6mm), protective column: (5μm, 12.50mm×4.6mm); mobile phase: methnaol-ultrapure water (80: 20); detective wavelength was at 270a good linearity within the range of 1×10-2-0.16μg, r=0.9995. The average recovery was 99.73% and RSD was 2.91%. CONCLUSION: The method is feasible, and simple to operate and suitable for the quality control of Compound Dangguifukang Capsule.

Methanotrophs could degrade methane and various chlorinated hydrocarbons. The analysis on methane monooxygenase gene cluster sequence would help to understand its catalytic mechanism and enhance the application in pollutants biodegradation. The methanotrophs was enriched and isolated with methane as the sole carbon source in the nitrate mineral salt medium. Then, five chlorinated hydrocarbons were selected as cometabolic substrates to study the biodegradation. The phylogenetic tree of 16S rDNA using MEGE5.05 software was constructed to identify the methanotroph strain. The pmoCAB gene cluster encoding particulate methane monooxygenase (pMMO) was amplified by semi-nested PCR in segments. ExPASy was performed to analyze theoretical molecular weight of the three pMMO subunits. As a result, a strain of methanotroph was isolated. The phylogenetic analysis indicated that the strain belongs to a species of Methylocystis, and it was named as Methylocystis sp. JTC3. The degradation rate of trichloroethylene (TCE) reached 93.79% when its initial concentration was 15.64 μmol/L after 5 days. We obtained the pmoCAB gene cluster of 3 227 bp including pmoC gene of 771 bp, pmoA gene of 759 bp, pmoB gene of 1 260 bp and two noncoding sequences in the middle by semi-nested PCR, T-A cloning and sequencing. The theoretical molecular weight of their corresponding gamma, beta and alpha subunit were 29.1 kDa, 28.6 kDa and 45.6 kDa respectively analyzed using ExPASy tool. The pmoCAB gene cluster of JTC3 was highly identical with that of Methylocystis sp. strain M analyzed by Blast, and pmoA sequences is more conservative than pmoC and pmoB. Finally, Methylocystis sp. JTC3 could degrade TCE efficiently. And the detailed analysis of pmoCAB from Methylocystis sp. JTC3 laid a solid foundation to further study its active sites features and its selectivity to chlorinated hydrocarbon.
Methylocystaceae ; classification ; metabolism ; Multigene Family ; Oxygenases ; genetics ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; Sequence Analysis, DNA ; Trichloroethylene ; metabolism

Objective To explore the effect of endothelial progenitor cells (EPCs) on the proliferation of co-cultured rat vascular smooth muscle cells(VSMCs). Methods Mononuclear cells were isolated from fresh cord blood by 6% hydroxyethyl starch (HES) and density gradient centrifugation. Isolated mononuclear cells were cultured in EGM-2 medium supplemented with 20% fetal bovine serum (FBS), VEGF,bFGF and other growth factors. Biological features of EPCs were observed at different time points, and EPCs were identified by morphology, fluorescence double-staining and flow cytometry. Indirect immunofluorescence was performed to analyze the expression of α-SM-actin, calponin of VSMCs special antigen. Co-culture system of EPCs and VSMCs was established by transwell permeable support. FBS (20%) was used to stimulate the proliferation of VSMCs. In a VSMCs/EPCs co-culture system, the DNA synthesis ability, total protein level and cell cycle of VSMCs were determined by BrdU marking method,protein quantitation and flow cytometry after co-culture for 6, 12, 24,48 and 72 h. Results After co-culture for 12, 24, 48, and 72 h, the DNA synthesis ability and total protein level of VSMCs significantly decreased compared with the control group(P＜0.05). Flow cytometry showed that the percentage of S phase of VSMCs in VSMCs/EPCs co-cultured group significantly decreased and the percentage of G_1 phase increased markedly compared with the control group(P＜0.05). The maximal inhibitory effect was observed at 48 h. Conclusion Early EPCs could inhibit the proliferation of VSMCs.