This study was aimed to establish a HPLC method for the content determination of liposoluble components, such as dihydrotanshione I, croptotanshinone, tanshinone I and tanshinone IIA, as well as the content determination of water-soluble components, such as danshensu, protocatechuic aldehyde, rosmarinc acid, salvianolic acid B, and salvianolic acid A in Danshen capsule simultaneously. For liposoluble components, the determination was performed on a Welch ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution using acetonitrile-water as the mobile phase. The flow rate was 1 mL·min-1. And the detection wavelength was set at 270 nm. For water-soluble components, the determination was performed on a Thermo Syncronis C18 column (250 mm × 4.6 mm, 5μm) by gradient elution using methanol-acetonitrile-0.02%phosphoric acid as the mobile phase. The flow rate was 1 mL·min-1. And the detection wavelength was set at 286 nm. The results showed that there were good linear relationships between components of peak areas and the ranges were 0.472-9.44 μg (r = 0.999 8) for danshensu, 0.352-7.04 μg (r = 0.999 9) for protocatechuic aldehyde, 0.244-4.88 μg (r = 1.000 0) for rosmarinc acid, 2.268-45.36 μg (r = 0.999 9) for salvianolic acid B, 0.168-3.36 μg (r = 0.999 9) for salvianolic acid A, 0.088-1.76 g(r=0.999 9) for dihydrotanshione I, 0.18-3.6μg (r=0.999 9) for croptotanshinone, 0.208-4.16μg (r=0.999 9) for tanshinone I, and 0.17-3.4μg (r=0.999 9) for tanshinone IIA. Average recoveries of the method were between 97.48%and 98.59%. It was concluded that the analysis was stable and reproducible, which can be used as a method for the analysis of Danshen capsule.

To retrospective analyze the clinical profiles of 80 patients undergoing thoracotomy with protection of intercostal nerve versus traditional method.The doses of narcotics of two groups were (12 ± 5)and (43 ± 11) mg respectively.The postoperative levels of visual analogue score (VAS) and such potential complications as pneumonia,atelectasis and paraesthesia were examined (P ＜ 0.01).Protective technique of intercostal nerve during thoracotomy could effectively relieve postoperative chest pain,reduce the dosage of narcotics and lower the occurrence of lung complications.

Objective:To study the effect of Liuwei Dihuang decoction (LW) on secretion of testosterone in the testis leydig cells of senescence accelerated mouse (SAM). Methods:The level of testosterone in the testis leydig cells of SAM with aging and the effect of LW on the secretion of testosterone were observed using cultured testis leydig cells in vitro.Results:The level of testosterone in the testis leydig cells of SAMP8 with aging was significantly decreased and showed a significant difference compared with age-matched SAMR1. Chronic administration of LW (2.5 and 5.0 g/kg) for 5 months significantly ameliorated the secretion of testosterone in SAMP8 compared with control group. Conclusions:The secretory function of testis leydig cells was reduced in SAMP8 with aging. LW ameliorated the secretion of testosterone in the testis leydig cells,indicating that LW could antagonize or delay the deterioration of the testis leydig cells in SAMP8.

Objective To clone the human target gene HBVDNAPTP1 transactivated by hepatitis B virus DNA polymerase obtained by screening with suppression subtractive hybridization (SSH) and bioinformatics techniques. To construct prokaryotic expressive vector of HBVDNAPTP1 gene, induce the expression of recombinant protein in E. coli, and analyze the expression level of HBVDNAPTP1 gene in human tissues. Methods The DNA fragment of HBVDNAPTP1 was amplified by reverse transcription polymerase chain reaction (RT-PCR) taking mRNA from HepG2 cells as the template, and the correct DNA fragment was then inserted into inducible prokaryotic expressive vector pET-32a (+). The competent BL21 (DE3) E. coli was transformed, and then cultured and induced with IPTG. The expressed HBVDNAPTP1 was confirmed with Western blot. UniGene database was used to analyze the chromosome mapping and tissue expression profile of HBVDNAPTP1 gene. Results The DNA fragment of HBVDNAPTP1 was amplified by RT-PCR. HBVDNAPTP1 expressive vector was constructed. After transformation with pET-32a(+)-HBVDNAPTP1 and induction with IPTG, recombinant HBVDNAPTP1 was expressed and confirmed by Western blot. The expression of genomic location of HBVDNAPTP1 gene was low in multiple-tissues with the exception of pituitary gland, tonsil, tongue, thymus, trachea and umbilical cord. Conclusion The recombinant HBVDNAPTP1 gene could be expressed in prokaryotic expression system of E. coli. The chromosome mapping and tissue expression level of HBVDNAPTP1 gene is tentatively conceived.

Objective To study the exact function of HBeBP4A so as to investigate the gene expression of HBeBP4A in yeast cell.Methods Reverse transcription polymerase chain reaction(RT-PCR)was employed to amplify the gene of HBeBP4A from recombinant plasmids pcDNA 3.1/myc-HisA-HBeBP4A,and the gene was cloned into pGEM-T vector.The gene of HBeBP4A was cut from pGEM-T-HBeBP4A vector and cloned into yeast expressive plasmid pGBKT7,and pGBKT7-HBeBP4A was then transformed into yeast AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting hybridization.Results HBeBP4A gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The SDS-PAGE and Western blotting assay showed that the relative molecular weight of the expressed product was about 61.37kD,and HBeBP4A protein existed in yeast cells.Conclusion The findings suggested that HBeBP4A was successfully expressed into yeast system.

Objective [WT5”BZ] To study the relationship between human papillomavirus (HPV) 16 and its serum antibody in the development of gastric carcinoma (GC) [WT5”HZ] Methods [WT5”BZ] Polymerase chain reaction (PCR) technique was used to detect HPV DNA in 42 fresh GC specimens and 42 fresh normal gastric mucosa adjacent to the tumour (NGMAT). Enzyme linked immunosorbent assay (ELISA) was used to screen the serum HPV16 antibody(Ab) of 42 GC patients and 46 controls using HPV16 virus like particles (VLPs) which was produced by recombinant bacilovirus in insect cells [WT5”HZ] Results [WT5”BZ] HPV16 DNA was found in 26 2% (11/42) of GC specimens,but in none (0/42) of NGMAT ( P

Objective To construct a cDNA subtractive library of genes transactivated by c-terminally truncated middle surface protein of hepatitis B virus(MHBs t) with suppression subtractive hybridization technique for cloning genes associated with transactivation. Methods The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-Mt167 and pcDNA3.1(-) empty vectors, respectively, then cDNA was synthesized. After restriction enzyme Rsa I digestion, small-size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. Results The subtractive library of genes transactivated by MHBs t was constructed successfully. The amplified library contained 94 positive clones. Colony PCR showed that these clones contained 200-800bp inserts. Sequence analysis was performed in 50 clones,and the full length sequences were obtained with bioinformatics method. 23 coding sequences were obtained in total, which consisted of 19 known and 4 unknown ones.Conclusions The obtained sequences may be target genes transactivated by MHBs t, among which some genes coding proteins may involve in cell cycle regulation, immune response and tumour genesis.

Objective To evaluate the importance of clinical screening and follow up by direct colonoscopy for colorectal cancer at an early and curable stage. Methods There were 2 196 elderly people aged between 60 to 89 years. The clinical screening by direct colonoscopy was performed according to the protocol. 1 740 of 2 196(79.2%) patients were followed up every year. Results Fifty two elderly persons were found to be colorectal cancer patients by colonoscopy, with the detectable rate being 2.4%. Nineteen were diagnosed early stage colorectal cancer, accounting for 36.5% of the detected colorectal cancer. Nine among the followed up cases were detected early colorectal cancer, accounting for 45 0% of the detected colorectal cancer. The resectable rate and the 5 year survival rate was 97 7% and 80 9% for colorectal cancer, respectively. 98 9% of the cecum intubation cases was successful. The incidence of complication for colonoscopy was 0 05%. Conclusions By clincal colonscopy screening and follow up study for colorectal cancer and precancerous changes in the elderly, the patients with adenomatoid polyps were early diagnosed and treated, so it raised the detectable rate of early colorectal cancer and the level of grade prevention of colorectal cancer.

Objective:To observe effects of Shenkui Decoction on proliferation of ovarian cancer cell.Methods:The Shenkui Decoction- containing rat serum was prepared by using TCM serum pharmacological method,and effects of the drug containing serum on proliferation of ovarian cancer fresh parenchymatous tumor cells were investigated with 3H incorporation method,and effects of the drug-containing serum on proliferation of ovarian cancer cell line Tyk-nu cells were investigated by flow cytometry,cellular activity assay,cellular growth curve assay and cell colony formation rate assay.Results:The drug containing serum could inhibit proliferation of both fresh parenckymatous tumor cells and ovarian cancer cell line Tyk-nu cells,and the inhibitory rate raised with the increase of drug-containing serum content.Conclusion:The Shenkui Decoction-containing rat serum can inhibit proliferation of ovarian cancer fresh parenchymatous tumor cells and ovarian cancer cell line Tyk-nu cells.

Objective To explore the feasibility and reliability to build a sinoatrial node damage model in canine induced by formaldehyde wet dressing.Methods Twenty dogs were randomly divided into 4 groups(5 in each group) by means of random number table:2-hour,24-hour,1-week and 4-week groups after wet dressing.The sinoatrial node area of canine was damaged by wet dressing with 20% formaldehyde.Sinus node function was measured before,2 h after wet dressing and corresponding times of each group.Electrocardiogram(ECG) of body surface were recorded synchronically.Results Average wet dressing time was 6.2?2.6(3 to 12)min.Five dogs showed significantly decrease of heart rate(HR)(140?11 vs 89?6 beat/min,P