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.

BACKGROUND: Researches on the pathogenesis and pathological changes of rheumatoid arthritis(RA) have achieved significant progress in recent years. But traditional Chinese medicine(TCM) has unique advantage in RA therapy.OBJECTIVE: To study effects of overall alkali in tongbiling(TBL) on the proliferation of lymphocytes and the transferrin receptor of T lymphocytes (CD71) to explore the mechanism of TBL on the modulation of cell immunity.DESIGN: A completely randomized grouping design and an explorative study by employing cells as subjects.SETTING: Sixth internal medicine department of a TCM university,center of tissue transplantation and immunology college of life science of a university PARTICIPANTS: The study was conducted in the central laboratory (tertiary laboratory of National TCM Administrator) of the first affiliated hospital of Guangzhou TCM medical university between July 2002 and August 2003. Ten clean male SD rats were selected.METHODS: Lymphocyte was separated from rat inguinal lymph node for culture. Concanavalin(ConA) was used for 72-hour stimulation. The impacts of overall alkali TBL on lymphocyte proliferation were tested by MTT.The expression of T lymphocyte CD71 was tested by flow cytometer after 48-hour stimulation of phorbol 12,13 -dibutyrate(PDB) or ConA.MAIN OUTCOME MEASURES: The impacts of overall alkali TBL on lymphocyte proliferation and T cell activation.RESULTS: Different concentration of overall alkali TBL could significantly inhibit the proliferation of lymphocytes under ConA stimulation. PDB and ConA-activated T lymphocyte CD71 + expressions were significantly higher than that of blank control group(P<0.01) . CD3+ CD71 + expressions [(62.03±1.51) %,(25.28±1.57) %,(20. 29±1.72)%] activated by ConA under different concentration of overall alkali TBL(50,100,200 mg/L)were significantly lower than(72.03±1.28)% of BPS-positive control group (P<0. 05). CD3 + CD71 + expressions activated by PDB under 100 mg/Land 200 mg/L of overall alkali TBL were significantly lower than that of phosphate buffer solution (PBS-)positive control group(P<0.05). Different concentration of overall alkali TBL had significant down-regulated effects on CD71 expression in T lymphocyte activated by PDB or ConA and there was also a significant dose-effect relationship(P<0. 05). The inhibition on ConA-activated CD71 expression was stronger than that of PDB.CONCLUSION: Overall alkali TBL can inhibit the abnormal proliferation of T lymphocyte and its mechanism might be realized through its inhibition on transferrin receptor.

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.

Chickens were infected with chicken anemia virus (CAV) at one-day-old and vaccinated with La Sota vaccine 8 days later. Meanwhile, uninfected chickens were vaccinated as controls. At 7, 14 and 28 days post vaccination, the content of IgG,IgM,IgA and HI titer in serum, the number of T cells, IgG, IgM and IgA antibody producing cells in thymus, bursa and spleen, the proliferative response of T、B cells, the inductive activity of interleukin 2 (IL-2) and interferon (IFN) in thymus and spleen were tested. The results showed that the content of IgG, IgM, IgA and hemoagglutination inhibition (HI) titer in serum, the number of T cells, IgG, IgM and IgA antibody producing cells in thymus, bursa and spleen, the proliferative response of T cells and B cells as well as the inductive activity of IL-2 and IFN in thymus and spleen of infected-vaccinated chickens significantly decreased compared with the control. These results indicated that the immunofunction and immunoregulation were dropped post ND vaccination of CAV-infected chickens.