[Objective]To research the efficient way of isolating and proliferating of induced endothelial cells(ECs) ,measure the number of the ECs and observe the main biological characteristic of ECs m vitro during the passage cultivation.[Method]The marrow mononuclear cells(MNCs) in rabbit marrow were isolated by the gradiem centrifugation and cultivated to isolate and induce ECs. The efficiency of isolating and harvesting ECs was measured by the number of the first passage ECs from every 10~6 of the MNCs. The changes of ECs morphology, growth and proliferation were observed during the passage cultivation. The immunohistochemical staining of CD31 and vWF and function of ECs were checked.[Result]After cultivated at 1?10~6/cm~2 MNCs for 7～8 d, the first passage ECs were harvested. The number of the first and the second passage ECs had a remarkable statistical positive correlation with the number of the MNCs from one rabbit. After cultivated for 7～8 d, the first passage ECs of 7.086?0.75?10~4 in number and the second passage ECs of 53.20?6.47?10~4 in number were harvested from every 10~6 MNCs on average;ECs, which were monolayer arrayed to form cobblelike shape,The cells of 2nd passage showed positive staining of CD31 and vWF, and positive labeling of DiI-LDL and UEA.[Conclusion]The period from cultivating to harvesting ECs is shortened at the density of 1?10~6/cm~2 MNCs. It is feasible that the efficiency of isolating and harvesting ECs could be measured by the number of the first or the second passage ECs from every 10~6 MNCs. The induced ECs can act as seeding cells in bone tissue engineering in future.

Objective: To raise the all-round informational level through analyzing the necessity of constructing compound medical library.Methods: Through aiming at the fundamental characters of compound medical library,so as to carry out the practices of theoretical researches,the development and optimization of network environments,the integration of informational resources,the enhancement and innovation of information service quality.Results: To provide some helpful references from the traditional to the compound state.Conclusion: Constructing compound medical library by combining the traditional with the digital base is the only way to accelerate the development of medical library.

Objective To establish a multiplex polymerase chain reaction (M-PCR) assay to detect HBV DNA in the peripheral blood mononuclear cells(PBMCs) of chronic HB patients. Methods One pair of primer amplifying HBV genome DNA and another pair of primer amplifying HBV covalently closed circular DNA (cccDNA ) were added to one PCR reaction to detect HBV DNA in PBMCs. Results Various forms of HBVDNA including total DNA and cccDNA could be amplified simultaneously. Among the 30 chronic HB patients, both the HBVDNA and HBVcccDNA in the PBMCs of 23 patients were detected, the positive rate of which was 76.6%. The positive rate of HBV cccDNA accounted for 82.1% of total HBV DNA positive rate. Conclusion HBVDNA in the PBMCs could partially replicate. The M-PCR was successfully set up to amplify HBV genome DNA and HBV cccDNA simultaneously.

The author analyses the chance and challenge of foreign periodicals booking in hospital libraries following China entering WTO and under the network environment,and proposes some effective solutions.

Objective:To observe the stability and sensitivity of transcription mediated ampliifcation (TMA) system, and to compare it with real-time reverse transcription polymerase chain reaction (RT-PCR) in amplifying serum HCV RNA in HCV infected patients. Methods: TMA system was established by moloney murine leukemia virus (MMLV) reverse transcriptase, T7 RNA polymerase and 2 speciifc primers ifrstly,and then its stability and repeatability were compared at different storage temperatures by the correlation change of HCV RNA amplification curve. The sensitivity difference between TMA and RT-PCR was evaluated by amplifying a group of 10-fold diluted HCV RNA samples which were transcribed in vitro. A total of 101 serums of chronic HCV infected patients were measured by TMA system and RT-PCR to observe the positive rate and their correlation. Linear correlation and linear regression were used to observe the correlation of the two methods. Results:TMA system was successfully established. TMA system was not stable when stored at 20℃ (placed for 24 hours only), but it was stable for 6 days when stored at 4℃ or within 6 months when stored at-20 ℃. Compared with RT-PCR whose reagent was made by Hunan Sansure Biotechology Corporation, TMA system showed 20 positive samples and 11 negative samples in a total of 31 samples. So was the RT-PCR kit of the Sansure Biotechology Corporation, and the concordance rate of the two methods was 100%. Advanced quantitative study of the 20 positive samples found that the two methods had good correlation and consistency (r=0.91,P<0.01). Compared with the results of RT-PCR whose reagent was made by Shanghai Kehua Bio-engineering Corporation, TMA system had 34 positive samples and 36 negative samples, while the RT-PCR technology had 32 positive samples and 38 negative samples out of 70 samples. hTe concordance rate of the two methods was 97.1%, with no statistical difference in the positive rate of the two methods (P>0.05). Advanced quantitative study of 29 positive samples found that the two methods had good correlation and consistency (r=0.96,P<0.01). Conclusion:The stability and repeatability of TMA system are good within 6 months when stored at-20 ℃ storage temperature. Both TMA and RT-PCR HCV RNA can detect serum HCV RNA well, and the two methods have good correlation and consistency.

ObjectiveTo study the relation ship between progressive cerebral infarction and thyroid hormone level,and analyze the risk factors of progressive cerebral infarction.MethodsThe thyroid hormone,triglyceride esters and fasting glucose levels of 48 patients with progressive cerebral infarction,138 patients with non-progressive cerebral infarction and 60 healthy subjects as control group were measured.ResultsCompared with the patients with non-progressive cerebral infarction,serum T3 of patients with progressive cerebral infarction was lower( P ＜0.05),and the levels of triglyceride ( TG),C-reactive protein ( CRP),and fasting blood glucose were higher ( all P ＜ 0.01 ).ConclusionPatients with progressive cerebral infarction were in a low level of thyroid hormones,and the increasing levels of TG,CRP,and fasting blood glucose were risk factors for progressive cerebral infarction.

It is an inevitable development tendency to build medical digital libraries. Our practice and results of digitalization of medical libraries explains the goal and idea of the construction of medical digital libraries.

To determine CD4 + CD25 + T regulatory cells (Tregs),forkhead box P3 (FoxP3) mRNA expression and levels of cytokines secreted by peripheral blood mononuclear cells (PBMCs) in individuals responsive or unresponsive to hepatitis B (HB) vaccination,and to explore the relationships between immune response and immune regulatory cells or cytokines.Methods Based on the antibody against hepatitis B surface antigen (HBsAg) after HB vaccination,the CD4 + CD25 + Tregs frequencies in PBMCs from 18 responders,22 nonresponders and 10 non-immunized healthy controls were analyzed by flow cytometry.The expression of FoxP3 mRNA in PBMCs with or without stimulation of phytohemagglutinin ( PHA ) and HBsAg was analyzed by real-time quantitative PCR.Levels of IL-4,IL-12,IL-18,and IFN-γ secreted by PBMCs after PHA and HBsAg stimulation were analyzed by enzyme-linked immunosorbent assay.Results The ratio of CD4 + CD25 + Tregs to CD4 + T cells in the nonresponders was markedly higher than that in the responders (P ＜0.05),but lower than that in the controls (P ＜0.01 ).FoxP3 was differentially expressed among the responders,nonresponders,and controls in PBMCs before and after PHA and HBsAg stimulation,and nonresponders had the highest FoxP3 mRNA expression (P ＜ 0.05 or P ＜ 0.01 ).The content of IFN-γ by PBMCs after PHA and HBsAg stimulation was markedly lower in the nonresponders as compared with the controls and responders (P ＜ 0.05).However,there were no significant differences in the levels of IL-18,IL-4,and IL-12 from PBMCs after PHA and HBsAg stimulation between the responders and controls as well as the nonresponders (P ＞ 0.05 ).Conclusion CD4 + CD25 + FoxP3 + Tregs may be involved in the negative regulation of responses to hepatitis B vaccination.Immunologic non-responses to hepatitis B vaccination may be related to IFN-γhyposecretion in PBMCs.

Objective Toinvestigate the changes of liver and kidney function and morphology of the renal artery of the metabolic syndrome (MS) rats.Methods The rats were respectively fed with ordinary or high-fructose diet for 10 weeks,and then took the blood and anatomical observation,to explore the changes of liver and kidney function and morphology of the renal artery of the rats.Results The serum levels of ALT,AST,UA,BUN,CR in the control group and MS group were gradually increased with the feeding time.But those in MS rats were significantly increased than the control rats.MS rats showed dysfunction of liver and kidney,which similar to the clinical MS cases.Compared with the control group,the renal artery of MS rats had the same pathological changes as MS cases,presented with intima uneven thickening and structural disorder,medial smooth muscle cells hyperplasia to intimal,the internal elastic membrane with different degrees of damage and fracture,the film elastic fibers thickening and structural disorder,smooth muscle cell proliferation apparently,medial obvious thickening.Conclusion MS rats have mild dysfunction of liver and kidney and abnormal changes in intima and tunica media of renal artery.

Objective To explore the correlation between N-terminal pro-brain natriuretic peptide ( NT-porB-NP) and D-dimer in coronary artery disease patients with type 2 diabetes .Methods 50 coronary heart disease patients with type 2 diabetes were selected as the study group ,and 30 healthy subjects at the same period were select-ed as the healthy control group .The serum levels of NT-porBNP and D-dimer were detected in two groups ,and the cor-relation between NT-porBNP and D-dimer was analyzed .Results The serum D-dimer and NT-porBNP levels in the study group were significantly higher than those in the control group [(2.43 ±0.58)mg/L vs (0.26 ±0.13)mg/L, (1 487.8 ±201.5)pg/L vs (83.3 ±21.4)pg/L](t=21.34,32.12,all P<0.05).The D-dimer level was positively correlated with the NT-porBNP level (r=0.564,P<0.05).Conclusion In coronary heart disease patients with type 2 diabetes ,the serum levels of D-dimer and NT-proBNP are high and closely related .