BACKGROUND: Aldosterone is an important pathogenic factor of left ventricular hypertrophy. There has been evidence that the extract of red sage root (a Chinese herb) can intervene the synthesis of aldosterone.OBJECTIVE: To investigate the effects of tanshinone Ⅱ A on expression of genes related to aldosterone synthesis in myocardium. DESIGN:A randomized and controlled trial. SETTING :Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS:The experiment was performed at the laboratory of Emergency Department, Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology from November 2002 to March 2004. Totally 20 male 12-week-old rats with spontaneous hypertension were randomly divided as hypertension group and tanshinone Ⅱ A group. METHODS:Rats in each group were injected tanshinone Ⅱ A or distilled water in the same volume respectively through caudal vein. After 12-week administration,the rats were put to death by decapitation, and the samples of myocardium were prepared. The expressions of CYP11B1mRNA and CYP11B2 mRNA, or myocardial genes related to aldosterone synthesis were examined with reverse transcriptase polymerase chain reaction (RTPCR) and referring to the amplification primers of glyceraldehydes 3-phosphate dehydrogenase (GAPDH) gene. MAIN OUTCOME MEASURES:The levels of CYP11B1 and CYP11B2 mRNA related to aldosterone synthesis in myocardium.RESULTS :Totally 20 rats were involved in the trial and all entered in the final result analysis without any loss of value. ① Quantitative analysis of expression of myocardial CYP11B1 and CYP11B2 genes in rats of two groups: Taking 100bp Plus Ladder as Marker,clear amplified strands could be seen at 440bp,461bp and 336bp sites,DNA sequencing proved they were the encoding gene segments of CYP11B1,YP11B2 and GAPDH. ②Qualitative analysis of expression of myocardial CYP11B1 and CYP11B2 genes in rats of two groups:The expressive levels of CYP11B1 and CYP1 1B2 mRNA contents in tanshinone Ⅱ A group were significantly lower than those in hypertension group (0.924±0.121 vs 1.343±0.132,P ＜ 0.05;1.017±0.119 vs 1.675±0.126,P 0.01). CONCLUSION:Tanshinone Ⅱ A could directly down-regulate the expression of CYP1 1B1 and CYP1 1B2 mRNA, the genes related to aldosterone syn thesis, so that it inhibited the biological synthesis of aldosterone in myocardinm, thus playing a role to resist hypertensive left ventricular hypertrophy.

BACKGROUND:Aldosterone is main pathogenic factor of left ventricular hypertrophy, for which, to inhibit its biosynthesis effectively may prevent and treat hypertension induced by left ventricular hypertrophy.OBJECTIVE: To investigate the effects of Tanshinone Ⅱ A on myocardiac aldosterone synthesis and expression of its relevant genic CYP11B1and CYP1 1B2 in spontaneously hypertensive rats(SHR) and explore the possi ble nechanism of Tanshinone Ⅱ A on inhibiting hypertension induced by left ventricular hypertrophy.DESIGN:SHR were employed as the objects in the experiment and WKY rats with normal blood pressure (WKY rats) were employed in the controlgroup. Complete group division and randomized control experiment was designed. Analysis of variance was used for the means comparison among groups.SETTING :Department of Emergency and Institute of Integrated Traditional Chinese and Western Medicine in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS :The experiment was performed in the laboratory of Department of Emergency in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 2003 to February 2004, in which, 10 male WKY rats with normal blood pressure of 12-week-old were employed in the control group and 20 male SHR of 12-week-old were randomized into two groups, named hypertension group and Tanshinone ⅡA group, 10 rats each group.METHODS:In Tanshinone Ⅱ A group, Tanshinone Ⅱ A 1.5 mg/kg was injected abdominally everyday. The distilled water of same volume was injected abdominally in hypertension group and the control group. Twelve weeks after experiment, myocardial specimens were collected after rats sacrificed. Radioimmunoassay was used to determine the contents of aldosteron and angiotensin Ⅱ in myocardial tissue and reverse transcriptase polymerase chain reaction (RT-PCR) were used to measure mRNA level of CYP11B1 and CYP11B2 genes relevant to aldosterone synthesis.MAIN OUTCOME MEASURES: Contents of aldosterone and angiotensin Ⅱ in myocardial tissue and mRNA expression of CYP11B1 and CYP1 1B2 genes.RESULTS:All of 20 SHR and 10 WKY rats entered result analysis. ① Myocardial aldosterone content of rats in 12 weeks after experiment: that in hypertension group and Tanshinone Ⅱ A group was higher remarkably than that in the control group [(0.056±0.014), (0.031±0.010), (0.018±0.009) ng/g,P ＜ 0.01, 0.05]; that in Tanshione Ⅱ A group was lower remarkably than that in the hypertension group (P ＜ 0.05). ② Myocardial angiotension Ⅱ content of rats in 12 weeks after experiment: that in the hypertension group and Tanshinone ⅡA group was higher remarkably than that in the control group [(0.093±0.016), (0.088±0.024), (0.043±0.012) ng/L, P ＜ 0.01]. ③ Expression of CYP11B1 gene in myocardial tissue of rats in 12 weeks after experiment: that in the hypertension group and Tanshinone Ⅱ A group was higher remarkably than that in the control group (2.774±0.138, 2.533 ±0.127, 1.973±0.102, P ＜ 0.05). ④Expression of CYP11B2 gene in myocardial tissue of rats in 12 weeks after experiment; that in the hypertension group and Tanshinone ⅡA group was higher remarkably than that in the control group (1.573±0.106, 1.024±0.113, 0.786±0.121, P ＜ 0.01,0.05); that in the Tanshinone ⅡA group was lower remarkably than that in the hypertension group (P ＜ 0.05). ⑤The electrophoresis band positions of myocardial CYP11B1 and CYP11B2 as well as RT-PCR products of glyceraldehydes-3-phosphate dehydrogenase (GAPDH), the consulting gene,were in conformity with the theoretic values.CONCLUSION:The inhibition of Tanshinone Ⅱ A on hypertension induced by left ventricular hypertrophy is probably contributed to its downregulating effect on myocardiac CYP11B2 gene expression relevant to aldosterone synthesis and to its reducing action on local biosynthesis of aldosterone in heart.

OBJECTIVE:To observe the effects of different concentration of human urinary kallikrein on blood pressure when used for acute cerebral infarction. METHODS: By a non-randomized historical control study,the blood pressure variation of 34 patients on day one after treatment with low concentration (0.06%) of human urinary kallikrein was observed,the occurrence of hypotension in this group was compared with that of another 47 patients (control) treated with high concentration (0.3%) of human urinary kallikrein. RESULTS: The reduction of blood pressure in low concentration group was less obvious than in the high-concentration group. However,hypotension was noted in both groups when angiotensin-converting enzyme inhibitor (ACEI) hypotensive drugs were used concomitantly. CONCLUSION: Human urinary kallikrein for acute cerebral infarction can transiently down-regulate blood pressure slightly,but its effect on blood pressure can be lowered by suitable reduction of its concentration;at any concentration,it can result in hypotension if used in combination with ACEI.

Objective To analyze the correlation between 18F-FDG SUV and immunohistochemical index including GLUT1, Ki-67, MVD, survivin and cyclinA in non-snall-cell lung cancer. Methods Thirty-three patients with NSCLC underwent preoperative PET/CT examination and surgical operation. All patients were divided into two groups according to the size of tumor (cutoff=3 cm), metastasis of mediastinal or hilar lymph nodes or not, and histological types of the cancer, respectively. The expression of GLUT1, Ki-67, MVD, survivin and cyclinA were estimated with SP immunohistochemical technique, and were analyzed statistically to reveal the correlation to FDG SUV. Results The rate of positive expression of GLUT1, Ki-67 and CD34 were 66.67%, 72.73% and 100%, respectively. The mean value of CD34 in all 33 patients was 12.6±2.9 (12-56). The rate of positive expression of survivin was 84.85%, and the corresponding data of cyclinA was 27.27%. Conclusion There is linear correlation between FDG PET SUV and GLUT1, but not between FDG PET SUV and Ki-67, MVD, survivin and cyclinA. The expressions of GLUT1, Ki-67, MVD, survivin and cyclinA are not related with the size of tumor, nor metastasis of lymph nodes. The expression of GLUT1 and Ki-67 is related with histological types of the cancer, but not with MVD, survivin and cyclinA.

Objective To investigate the efficacy and side-effect of docetaxel and cisplatin induction chemotherapy followed by concurrent chemoradiotherapy in locally advanced non-small cell lung cancer (NSCLC).MethodsEighty-six patients with histologically confirmed locally advanced NSCLC were randomized into induction chemotherapy followed by concurrent chemoradiotherapy (ICCRT)arm or concurrent chemoradiotherapy (CCRT) arm. Both arms were treated with intensity-modulated radiation therapy. Induction and concurrent chemotherapy regimen consist of docetaxel and cisplatin. Results Follow-up rate of the whole group is 100％.The response rate in the CCRT arm and ICCRT arm is 70％ and 80％ ( χ2 =1.26,P =0.261 ),respectively; and 1-,2-,3-year survival rate is 65％ and 85％,40％ and 50％,33％ and 44％ (χ2 =3.90,P=0.048),respectively; the median survival time and time to progression is 17.5 and 22.0 months and 14.0 and 19.0 months respectively.Major adverse effects are leukopenia (43 and 32 cases,χ2 =3.48,P =0.062),radiation esophagutis (26 and 20 cases,χ2 =0.12,P =0.730),anemia (26 and 16 cases,χ2 =2.34,P =0.126) and radiation pneumonitis (13 and 9 cases,χ2 =0.37,P =0.541 ).ConclusionsICCRT for locally advanced NSCLC can improve the overall survival rate and time to progression,induction chemotherapy did not increase side-effects.There was no difference in response rate between CCRT and ICCRT arm.

We engineered a disulphide bridge between two adjacent double-layered beta-sheet at the N-terminal region of Trichoderma reesei endo-1,4-beta-xylanase II(XYN II) by site-directed mutagenesis. The native xylanase XYN-OU and the mutated xylanase XYN-HA12 (T2C, T28C and S156F) were separately expressed in Pichia pastoris. Both xylanases were purified and characterized. The optimum temperature of XYN-HA12 was increased from 50 degrees C to 60 degrees C, relative to XYN-OU. At 70 degrees C, the halftime of inactivation for XYN-OU and XYN-HA12 were 1 min and 14 min, respectively. The optimum pH of XYN-HA12 was 5.0, similar to XYN-OU. However, XYN-HA12 could retain over 50% activity from pH 3.0 to 10.0 at 50 degrees C for 30 min. As for XYN-OU, it could retain over 50% activity from the pH value 4.0 to 9.0 at 50 degrees C in 30 min. The result of the mutated xylanase indicated that constructed disulphide bridge could improve its thermostability at relatively higher temperature.
Amino Acid Substitution ; Disulfides ; chemistry ; metabolism ; Endo-1,4-beta Xylanases ; biosynthesis ; chemistry ; genetics ; Enzyme Stability ; genetics ; Mutagenesis, Site-Directed ; Pichia ; genetics ; metabolism ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; chemistry ; Trichoderma ; enzymology ; genetics