Objective To observe the Cordyceps extract on experimental rats with diabetic kidney and to explore its mechanism of action.Methods The rat models of diabetic were established by intraperitoneal injecting streptozotocin.Kidney weight and the index of kidney were measured,and 1,4,7weeks blood sugar,serum creatinine,urine microalbumin and urea were dynamically observed for evaluating Cordyceps extract on experimental rats with diabetic kidney.Results The 1,4,and 7 weeks of kidney weight was (0.662± 0.062) g,(0.670±0.061) g,(0.657±0.063) g,renal index was (0.312±0.041) ％,(0.309±0.402) ％,(0.311 ±0.0317) ％,serum creatinine was (0.51±0.03)mg/d1,(0.57±0.03)mg/d1,(0.52±0.02)mg/d1,albumin was (1.77±0.17)mg/24 h,(1.52±0.19)mg/24 h,(1.56±0.11)mg/24 h.urea was (90.71 ±0.37)mmol/L,(91.57±0.48) mmol/L,(90.56±0.39)mmol/L in the normal control group respectively.While in the model group,the value of kidney weight was (0.879±0.037)g,(0.912±0.038)g,(0.871±0.393)g,renal index was (0.494±0.039)％,(0.487±0.038)％,(0.465±0.235)％,serum creatinine was (3.71 ±0.14) mg/d1,(3.51±0.12) mg/d1,(3.32±0.11)mg/d1,urinary albumin was (7.29±0.22)mg/24 h,(7.11±0.34)mg/24 h,(7.14±0.21)mg/24 h.urea was (109.59± 8.42),(92.52±2.41),(90.71 ±0.67) respectively.Compared with the normal control group these values were decreased significantly in the model group (P＜0.05).In the Cordyceps sinensis group kidney weight was (0.837±0.036)g,(0.747±0.029)g,(0.694±0.357)g,renal index was (0.412±0.024)％,(0.395±0.037)％,(0.361±0.027)％,serum creatinine was (1.44±0.11)mg/d1,(0.81± 0.04) mg/d1,(0.62±0.03) mg/d1,urinaryalbumin was (5.71±0.25)mg,(4.52±0.21)mg,(3.96±0.15)mg,urea was (109.59± 8.42) mmol/L,(92.52±2.41) mmol/L,(90.71 ± 0.67) mmol/L respectively.Compared with the model group these values were decreased significantly(P＜0.05) in the Cordyceps sinensis group.Conclusion Cordyceps sinensis extract has a protective effect on kidney of diabetic rats.

Objective The purpose of this study was to investigate the expression of basic fibroblast growth factor (bFGF) and its effect on tumor interstitial angiogenesis and the biological behavior of gastric carcinoma. Methods CD34 was used to explore the microvessel density (MVD) as the marker of endothelial cell. bFGF was qualitatively and quantitatively detected with immunohistochemistry on 74 cases of gastric carcinoma and 17 cases of adjacent normal gastric tissue. Results Measurement of CD34 expression could be used to describe endothelial cell proliferation and vascularity. Strong expression of bFGF was localized in the cytoplasm of tumor cell and interstitial new microvessel endothelial cells. Expression of bFGF in tumor tissue was stronger than in corresponding non tumor normal gastric tissue ( P

BACKGROUND: Streptococcus mutans (S. mutans) is generally considered to be the principal aetiological agent for dental caries, thrS gene may relate to the virulence of S. mutans involved in the adherence, acidogenicity and acidodurance.OBJECTIVE: To investigate the conservation status of the thrS gene of S. mutans and to construct the homologous recombinant plasmid.METHODS: Southern Blot was used to analyze the distdbuUon of thrS gene in S. mutans. The upstream and downstream sequences of thrS gene were cloned respectively into multiple cloning sites of suicide plasmid pFW5 to construct the recombinant plasmid,RESULTS AND CONCLUSION: The thrS gene was conserved in 6 strains of S. mutans in this test. By PCR analysis and enzyme digesting, it was confirmed that S. mutans thrS gene homologous recombinant plasmid was successfully constructed,which can be used in future research of construction of thrS -negative mutans of S. mutans strain UA159.

Objective To observe the effect ofDanhuai-Yinxie condensed pill on the expression of VEGF and its receptors in the model rabbits with psoriasis.Methods Fourty-two healthy Japanese white rabbits were randomly divided into the control group, the model group, the Acitretin group, and the low-, medium- and high-doses ofDanhuai-Yinxie condensed pill groups. The backs of rabbits’ ears were wiped with 5% propranolol for 4 weeks in the latter four groups, while with saline for the same period in the control group. The rabbits in the Acitretin group were intragastric administrated with 1% Acitretin one day since the models of psoriasis were successed, while the rabbits in the low-, medium- and high-doses ofDanhuai-Yinxie condensed pill groups were intragastric administrated withDanhuai-Yinxie suspension 20, 40, 80 mg/kg daily, respectively. The rabbits in the control and model groups were intragastric administrated with equal-volume saline daily. The expression of VEGF, its receptor 1 (FLT-1) and receptor 2 (KDR) were detected by ELISA method, and the expression of VEGF mRNA was detected by PCR method, dermal papillary vascular density (MVD) and the T lymphocytes proliferation from peripheral blood were detected.Results The expression of VEGF (114.43 ± 952 ng/ml, 109.65 ± 9.15 ng/mlvs. 179.90 ± 11.30 ng/ml), VEGF mRNA (0.83 ± 0.07, 0.80 ± 0.06vs. 2.74 ±  0.15), FLT-1 (4.54 ± 0.26 ng/ml, 4.16 ± 0.24 ng/mlvs. 6.92 ± 0.38 ng/ml), KDR (2.48 ± 0.16 ng/ml, 2.51 ± 0.19 ng/mlvs. 6.79 ± 0.27 ng/ml), MVD (10.12 ± 1.97, 9.83 ± 1.12vs. 16.45 ± 2.05) and T lymphocytes proliferation (87.35% ± 6.29%, 86.21% ± 6.42%vs. 123.74% ± 9.18%) in the medium- and the high-dose groups significant decreased than those in the model group (P<0.05).ConclusionDanhuai-Yinxiecondensed pill could reduce the expression of VEGF, block the binding of VEGF to its receptors, inhibit angiogenesis, inhibit the reconstruction of subcutaneous tissues, reduce the dysplasia of dermal microvessels, and inhibit the proliferation of T cells.

Objective:To knock the phospho-sugar mutase gene (psm) out of the genome of S. mutans strain UA159 and to construct the mutant will lay a foundation for the study of the function of the psm. Methods:Two upstream and downstream DNA sequences of the psm were selected and cloned respectively into multiple cloning sites I and II of suicide plasmid pFW5 to construct the recombinant plasmid which was confirmed by enzyme digesting and sequencing. The recombinant plasmid was transformed into S. mutans UA159 by natural transformation and antibiotic was used to screen the positive transformants. According to the principle of homologous recombination, allelic exchange between the recombinant plasmid and S. mutans UA159 was achieved. Results:By PCR analysis and sequencing, it was confirmed that the psm of S. mutans UA159 was substituted for resistant gene of spectinomycin. Conclusion: The psm-negative mutants of S. mutans UA159 is successfully constructed.

Objective:To construct the prokaryotic expression vector pET28a(+)-comE, to get the highest expression of comE in E. coli BL21(DE3), and to obtain the purified ComE fusion protein. Methods: ComE gene was amplified by PCR with specific primers from genome of Streptococcus mutans UA159. After digested by two restriction endonucleases BamH Ⅰand Xho Ⅰ, the gene segment was inserted into vector pET28a(+). Then the recombinant vector was transformed into E. coli BL21(DE3). The protein expression was induced by IPTG and the result was confirmed by Western blot. The solubility of the fusion protein was examined by 12% SDS-PAGE, and the expression conditions were optimized. Finally the fusion protein was purified by a two-step purification procedure utilizing Ni2+ chelating column and S75 size exclusion column. Results: The recombinant plasmid was confirmed by PCR, restriction endonucleases digestion and sequence analysis, which showed that the inserted segment was identical to comE gene reported by GenBank without nucleotide mutation. The fusion protein which was confirmed by Western blot can be expressed in E. coli BL21(DE3) and existed both in supernatant and precipitation. The maximum expression product amount was obtained when the A600 value of E. coli BL21(DE3) was 0.4, IPTG concentration was 0.10 mmol/L and induction time was 4 h. Conclusion: The recombinant plasmid is constructed and the ComE fusion protein is purified by the optimum conditions.

Objective To study the relation between changed atrial load and atrial septal motion.Methods Forty one patients with valve diseases,7 patients with congenital heart diseases, and 12 normal subjects were involved.Two dimensional image was obtained by transesophageal echocardiography.The thickness,wave time and velocity were showed during atrial septal motion by omni directional M mode echocardiography (OME).Results With increase of the volume load and pressure load in the atrium,the atrial septal thickness increased, and the velocity of atrial septal motion increased or reduced.Conclusions OME shows that the thickness and velocity of atrial septal motion relates with condition of atrial load.

Objective:To construct a suppression subtractive library of suppression-related genes from c serotype Streptococcus mutans (S.mutans). Methods:After being isolated from virulent and avirulent strain of S. mutans respectively, the intact and high-pure genomic DNAs were digested with restriction enzyme AluⅠ. The digested DNA of the avirulent strain ligated with an adaptor was used as tester DNA, and that of the virulent strain as driver DNA. Then the suppression subtractive hybridization(SSH) was carried out, the efficiency of ligation and subtraction were detected respectively. The subtracted fragments were inserted into vector pCR2.1 using T/A cloning kit and transformed into E. coli TOP10F' competent cells. The white colonies were selected to construct the suppression subtractive library. Results: Through electrophoresis of AluⅠ-digested DNAs, a smear ranged from 0.1 to 2.0 kb was observed. The ligation efficiency of tester DNA with adaptor was at least higher than 25 percent. The subtraction efficiency confirmed the success in enrichment of differential genes between virulent and avirulent strain of S. mutans. In the subtracted group, the appearance time of the 23S rRNA gene in both tester and driver DNA was later than that in the unsubtracted group by twelve cycles. It suggested that suppression subtractive hybridization happened indeed. Then the subtracted fragments were cloned and the suppression-related gene library between virulent and avirulent strain of S. mutans was constructed. Conclusions:The suppression subtracted library of suppression-related genes has been constructed.

Objective:To explore avirulent strain-specific new genes or new functions of already known genes from Streptococcus mutans of serotype c.Methods:Twenty-six DNA fragments unique to avirulent strain of Streptococcus mutans were sequenced.The sequences of these presumptive avirulence DNA fragments were subjected to homology search through BLASTn and BLASTx in public database,and their putative biological functions were analyzed.Results:The size of the DNA fragments ranged from 113 bp to 776 bp.The average G+C content was 38.27%,similar to that of the gene-codingsequences in Streptococcus mutans strain UA159 whose genome sequences were just completed.Seven clones were picked repeatedly.Of the nineteen DNA fragments,eight potentially represented new DNA fragments were registered in GenBank.The remaining DNA fragments showed high homology to known genes of Streptococcus mutans strain UA159.Conclusions: The gene analysis and identification supply the groundwork for further study of gene functions of the avirulent strain of Streptococcus mutans serotype c.

Objective To observe the effect ofShenmal injection on vascular endothelial function after cerebral ischemia reperfusion in rats.Methods 30 rats were randomly divided into a sham operation group, a model group and aShenmal group, with 10 in each. Except the sham operation group, animal models of cerebral ischemia reperfusion injury were established by the right internal carotid artery occlusion of middle cerebral artery. Then shenmal injection was given to theShenmal group, 2 ml/100 g Sodium Chloride was given to the other groups, once per day, for successive 7 days. Endothelin-1 (ET-1), nitric oxide (NO), nitric oxide synthase (NOS), serum high sensitivity C-reactive protein (hs-CRP), and homocysteine (Hcy) were tested. Results In Shenmal group, ET-1(126.10 ± 6.07 ng/mlvs.105.41 ± 0.09 ng/ml), hs-CRP (15.93 ± 2.79 mg/Lvs. 9.33 ± 1.32 mg/L) and Hcy (11.01 ± 0.98 μmol/Lvs. 8.13 ± 0.46 μmol/L) were decreased after the treatment, while NOS (168.87 ± 9.25 U/mlvs.242.25 ± 10.36 U/ml) and NO (84.03± 6.02vs.102.42± 5.05) were increased after the treatment with statistical difference(P<0.05).ConclusionShenmalinjection can improve NOS activity and promote the synthesis of NO, decrease the level of plasma ET-1, hs-CRP and Hcy, reduce endothelial inflammatory exudation, and prevent vascular endothelial dysfunction caused by cerebral ischemia reperfusion injury.