OBJECTIVE:To provide a new idea for enhancement of core competitiveness of pharmaceutical enterprises in China and supply advice for building virtual marketing network.METHODS:The concept and theory of virtual management and virtual marketing network were interpreted and analyzed according to the status quo of pharmaceutical enterprises in China.RESULTS & CONCLUSION:Virtual marketing network is a new idea for the development of pharmaceutical enterprises.Enterprises should establish a standard cooperative relation with the dealers in order to prevent risks and promote core competitiveness of enterprises.

ObjectiveTo evaluate the therapeutic effect of combined portoazygous devascularization and shunt on portal hypertension. MethodsThe data of 56 cases of portal hypertension treated with portoazygous devascularization and shunt from April 1987 to April 1999 was summarized. ResultsThere was no death in 54 patients receiving selective operation;of 2 cases receiving emergency operation, one died. Among 49 followed-up cases, there was one rebleeding,2 hepatic encephalopathy, and one died of liver failure 4 years after opeation. The mean portal pressure before and after operation was(3.42±0.46) kPa and (2.50±0.35) kPa, respectively. ConclusionCombined portoazygons devascularization and shunt not only decreases portal pressure but preserve hepatic blood flow to some extent.

Objective To investigate the change of homocysteine (Hcy),high sensitive C-reactive protein (hs-CRP)and Inter-leukin 6 (IL-6)in patients with Obstructive Sleep Apnea-Hypopnea Syndrome (OSAHS)complicated by Cerebral Infarction (CI).Methods Chose 55 patients with OSAHS and 56 patients with CI.All patients with OSAHS were divided into mild group,moderate group and severe group by polysomnography,and all patients were divided into simple OSAHS group,sim-ple CI group and OSAHS complicated by CI group.All the patients’sleep apnea hypoventilation index (AHI),the lowest oxygen saturation (LSpO2 ),average blood oxygen saturation (MSpO2 ),sleep apnea the longest (LAT)and the level of Hcy,hs-CRP,IL-6 were determined,and compared with 30 cases of control group.Results The AHI,LAT,Hcy,hs-CRP and IL-6 of simple OSAHS group and simple CI group were obviously higher than that in control group,LSpO2 ,MSpO2 were obviously lower,the differences were statistical significant (P<0.05).The AHI,LAT,Hcy,hs-CRP and IL-6 of OS-AHS complicated by CI group were obviously higher than that in simple groups,but LSpO2 and MSpO2 were obviously low-er,and the differences were statistical significant (P<0.05).With the aggravation of OSAHS,Hcy,hs-CRP and IL-6 were gradually increased.Hcy,hs-CRP and IL-6 were positively correlated with AHI in patients with OSAHS,correlation coeffi-cient were 0.751,0.678 and 0.635,respectively (P<0.001).Conclusion OSAHS was closely related to the occurrence and development of CI,the dynamic monitoring of Hcy,hs-CRP and IL-6 levels has a certain reference value for evaluation of therapeutic effect.

Objective To study the coexpression of protein folding modulators,including chaperones(DnaK,DnaJ,and GroEL-GroES) and disulfide isomerase Dsb molecules(DsbA and DsbC) on the bioactivity of eukaryotic expressed urokinase(UK).Methods The gene sequences of DnaK,DnaJ,GroEL-GroES,DsbA and DsbC were isolated from the chromosome of E.coli by PCR.The obtained fragments mentioned above were cloned into the plasmid pBAD-1 under the control of araB promoter and then the plasmids were transformed into E.coli BW25113 and HB101 together with a compatible vector pQE-UK.The amount of expressed UK was measured by Bradford method and its activity was analyzed by fibrin plate method for its fibrinolysis.Results The activity of expressed UK was increased by co-expressing with DnaK,GroEL-GroES,DsbA,and DsbC in BW25113.Among them,the highest activity was achieved by co-expressing with GroEL-GroES.Conclusion Co-expressing of chaperones or disulfide isomerase can improve the bioactivity of UK expressed in E.coli.

Objective To evaluate the feasibility of intra-utero fetal lamb surgical repair. Methods Six twin lambs underwent surgery intra-utero at 112 days of gestation (term 140～160 days). After maternal laparotomy and Hysterotomy, fetal lamb′s toe was excised or its cleft lip was repaired in one of twin. Results At 30 to 32 days post operation,five lambs were spontamcously delivered and the other was cesarean delivery to full term gestation. Fetal wounds healed without inflammation and scar formation. Conclusions The methods of fetal lamb intro utero surgical repair is feasible.

BACKGROUND: Rapid development of tissue engineering is the new hope of repairing bone defect so as to obtain complete bone regeneration,and rapid proliferation of seed cells is one of the important factors in it.OBJECTIVE: To evaluate the proliferation and osteogenic differentiation of human mesenchymal stem cells (MSCs) plated at different cell density during subculturing period.DESIGN: Repetitive measurement.PARTICIPANTS: The experiment was conducted in the Trauma, Burn and Combined Injury Laboratory of the Third Military Medical University of Chinese PLA between January 2003 and March 2004. MSCs were obtained from iliac bone bone marrow of the 5 healthy male volunteers by separation and proliferation.METHODS: The MSCs of second generation were inoculated at 8×103/cm2,3×103/cm2, and 8×102/cm2, respectively, and then the growth curve was drawn and analyzed. After 18 days of proliferation and culture, the increased amount of MSCs was recorded to analyze the post-proliferation growth curve and osteogenic ability.MAIN OUTCOME MEASURES: ①Growth curve of cells plated at different cell density; ②Results of alkaline phosphatase(ALP) staining and immunohistochemical staining of osteocalcin.RESULTS: Human MSCs from bone marrow were CD34 negative and expressed low level of ALP. Population doubling time was 40 hours after MSCs were plated at 8×103/cm2 and the number of the cells was expanded (51±13) times after 18 days. MSCs were expanded (28±6) times 18 days after plated at 3×103/cm2. While plated at 8×102/cm2, the number of the cells was only expanded (5±3) times. Proliferative ability of cells cultured at a low density was stronger that that at a high density. MSCs could be induced to differentiate into the osteocytic lineages after plated at not only 8×103/cm2 but also 8×102/cm2.CONCLUSION: Cell density plays an evident role in the proliferation of MSCs. Cell density of 8×103/cm2 is proper to the rapid proliferation of MSCs as seed cells in bone tissue engineering.

Objective To study the relationship between homocysteine (Hcy)and carotid atherosclerosis in patients with cer-ebral infarction.Methods During January and November 2013,281 patients with cerebral infarction from Affiliated Hospital of Yan’an University were provided the blood samples as cerebral infarction group,140 healthy volunteers served as control group.Serum Hcy was detected by enzymatic cycle assay.According to the results of carotid artery ultrasonography,cerebral infarction group was divided into five groups:artery normal group,intimal thickening group,soft plaque group,mixed plaque group and hard plaque group.The statistically significance was analyzed by SPSS 19.0.Results The serum Hcy level of cer-ebral infarction group was 19.78 ± 5.21 μmol/L,significantly higher than the control group 10.24 ± 3.33 μmol/L (P <0.001).The serum Hcy levels of control group,artery normal group,intimal thickening group,soft plaque group,mixed plaque group and hard plaque group were 10.24±3.33,15.20±2.99,17.03±1.85,25.44±4.24,19.65±4.74 and 18.31 ±3.67 μmol/L respectively.The differences between groups were statistically significant in addition to intimal thickening group and hard plaque group (P =1.106).The positive rates were 16.4%,53.7%,87.1%,95.7%,83.1% and 77.3% re-spectively,the groups which in cerebral infarction group compared with control group,the differences were statistically sig-nificant (P <0.001).Conclusion Hcy played an important role in the occurrence and development of cerebral infarction. Lower serum Hcy concentration,may be an effective way to prevent carotid atherosclerosis and cerebral infarction.

Objective:To study deeply on the effects of sinomenine on Th1 type cytokines expression of T lymphocytes.Methods:The authors isolated and cultured the mesenteric lymph nodes of mice in vitro.The lymphocytes were stimulated with PDB and inomycin after added sinomenine in different concentrations for 1 hour.After 4 hours stimulation,cells were collected and stained intracellular cytokines with the dying regents,then the expression of Th1 type cytokine IFN-?,pro-inflammatory cytokine TNF-? of T lymphocytes were analyzed by flow cytometry.And also observed the effects of drugs on the expression of CD69,an early marker of T cell activation.Results:1 000 ?mol/L(329.4 ?g/ml)sinomenine can downregulate CD69 expression of T lymphocytes while 200 ?mol/L sinomenie show no effect on it.200,1 000,2 000 ?mol/L sinomenine can obviously reduce the expression of intracellular cytokines such as IFN-?,TNF-? in dose-dependent manner.Conclusion:One of the immunological mechanisms of sinomenine to treat rheumatoid arthritis may due to the inhibitory effects on abnormal T lymphoctytes activation.The inhibitory effects of sinomenine may mainly be relative to the inhibitory effects on Ca 2+ -dependant signal pathway of T lymphocytes activation but not PKC pathway.

[Objective] To observe the effects of Tongbiling (TBL) prescription in counteracting cartilage destruction in rats with rheumatoid arthritis (RA). [Methods] Collagen-induced arthritis (CIA) rat models were established to observe the therapeutic effects of TBL. After a short-term treatment, expressions of tumor necrosis factor (TNF - a) and transforming growth factor (TGF- P) in cartilage tissue of model group, methotrenxate (MTX) tablets group and Radix Tripterygii Wilfordii (RTW) preparation group were examined by immunohistochemical semi-quantitative assessment method. [ Results ] The number of chondrocytes with positive TNF - a expression was lower and the chondrocytes were stained lighter in normal group than those in model group and low-dose MTX group ( P

[ObjectivejTo explore the therapeutic mechanism of Tongbiling (TBL) prescription for rheumatoid arthritis (RA) by observing the effects of TBL on inflammation and expression of vascular endothelial growth factor (VEGF) in synovium of rats with collagen-induced arthritis (CIA) .[Methods]CIA rat models were established by subcutaneous injection of type II collagen. Pathological sections of synovium in knee joint were made after medication for 37 days and then stained by HE method and immunohistochemical method respectively. Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in HE-stained pathological sections were observed under microscope and expression of VEGF was observed by semi-quantitative immunohistochemical method. [ Results ] Infiltration of inflammatory cells and hyperplasia of macrophages A and fibrous tissue in high-dose TBL group were inferior to those in model group (P 0.05); both TBL and Radix Tripterygii Wilfordii at high dose decreased the expression of VEGF (P