Polycentric governance theory of public goods provides a new approach for the goal of Basic medical care and health service for all.From the perspective of the public goods property of basic medical care and health service,the paper analyzed the predicament brought one-center supply mode,and designed the basic medical care and health service polycentric supply mode on the basis on the polycentric governance theory.On the basis,it proposed the polycentric supply synergetic development strategies.

Currently,the research on evaluation of scientific and technological competency at universities is at its preliminary stage.To build an index system suitable for evaluating the competency of local medical colleges is import for its development.We here analyzed the key elements affecting the biomedical competency at local medical universities.Then,we constructed an index system with the Delphi method for the evaluation.Finally,the paper made an empirical research of this index system,which suggested that the evaluation index system is scientific and reliable.

Objective To construct the eukaryotic expression vector of human PRKCB1 containing enhanced green fluorescence protein gene and transfer into human umbilical vein endothelial cells(HUVECs),then identify activity of expression protein.Methods The pReceiver-M29-PRKCB1 eukaryotic expression plasmid was constructed by frame amplified from pMD18-T-PRKCB1 plasmid.Then the recombinant plasmids were identified by enzyme analysis and DNA sequencing.According to optimized conditions,the eukaryotic expression plasmids were transfered into HUVECs and observed under fluorescence microscope.After that,transfection efficiency was calculated under random vision.The plasma membrane/cytosol ratio of fluorescence was calculated under confocal microscope.The translocation was identified.Results The gene sequence was completely consistent with that reported in GenBank.The enhanced green fluorescence protein was observed in HUVECs after 48 hours.Transfection efficiency was 18.6%?1.6%.The translocation was observed.Conclusion The eukaryotic expression plasmid is successfullyconstructed and transfered into HUVECs,the translocation was identified.It is a potential tool for screening HUVECs stably expressing human protein kinase C ?2 and isolating protein complex.

Objective To study the expression changes of matrix metalloproteinases (MMPs), protein kinase C (PKC) and nuclear factor-?B (NF-?B ) on the injured rabbit vascular smooth muscular cells (VSMC) transfected with tissue inhibitor of metalloproteinase-2 (TIMP-2) vector. Methods Lipofectin method was used to transfect TIMP-2 vector into VSMC, Western blot analysis to detect TIMP-2 peptides and zymography assay to determine MMPs. The activities of MMPs and NF-?B were detected by electrophoretic mobility shift assay (EMSA). The mRNA and protein expressions of PKC? were determined by RT-PCR and immunocytochemistry. Results The injured VSMC showed increased enzyme activity of MMP-2. There was very lower level expressions of PKC? and NF-?B in the normal VSMC but high in the injured VSMC. However, in injured VSMC transfected with TIMP-2 vector, the activity of MMP2/9 was suppressed and the expressions of PKC? and NF-?B decreased (P

Objective To construct the endothelial cell model with overexpressed human protein kinase C ?2(PKC?2) after high glucose inducement in order to study the function of human PKC?2. Methods The PRKCB1 gene was amplified from pMD18-T-PRKCB1 plasmid and then directly cloned into shuttle plasmid pDC315 to construct shuttle plasmid. Then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHGlox△E1,3Cre were cotransfected into 293 cells to construct recombinant adenovirus Ad-PRKCB1. The virus titer was calculated by TCID50. The Ad-PRKCB1 was verified by immunocytochemistry and RT-PCR. Ad-PRKCB1 was transfected into human umbilical vein endothelial cells (HUVECs) followed by the treatment of high glucose (25 mmol/L) for 96 h, and then the cells were observed by laser scanning confocal microscopy (LSCM). Results Restrictive endonuclease digestion and PCR result showed that the target gene was correctly cloned into shuttle plasmid. Cytopathic effect (CPE) was observed under inverted microscope through homologous recombination. The virus titer was 7.9?109 IU/ml. Immunocytochemical staining and RT-PCR indicated the expression of human PKC?2 in the transfected HUVECs. The translocation was observed under LSCM. Conclusion A recombinant adenovirus vector with human PKC?2 and the endothelial cell model with human PKC?2 overexpression by high glucose are constructed successfully.

OBJECTIVE:To discuss the measures for diversified development of pharmaceutical retail chain stores in Chi-na.METHODS:The current problems existing in pharmaceutical retail chain stores were discussed,and the measures were put forward on the basis of these problems.RESULTS&CONCLUSION:Brand diversification is the only route of development for pharmaceutical retail chain stores in China,to which diversification in drug sales is a most supplement,and creativity is the source of diversification.

Objective To investigate the identification of MiR-221 regulates apoptosis by targeting PUMA in lung cancer. Methods The express levels of miRNAs that collected from lung cancer tissue and lung benign lesion were tested by the particular kid they made,analyzed the correlation between individual miRNA.Results 9 6 possible regulated PUMA related miRNAs had been screened through the bioinformatics database,then customized it into the special kit to test the expression with real time fluorescent PCR.The results showed that 10 types miRNAs were up expression in lung cancer tissue,miR-NA221 were selected for deep analyze.Results with rank sum test showed statistical significance (P<0.05)in lung cancer and para cancer tissue.There were statistically significant differences in the expression between metastasis and nonmetastasis (P<0.05),but had nothing to do with the pathological types.Conclusion Real time fluorescent PCR test showed several types of PUMA related miRNAs up-expressed in tissue lung,which indicate miRNA have relationship with PUMA in gene express regulation and cancer generation and development.Pression situation and clinic pathological parameters.Expression of miRNA-221 in lung cancer was associated with tumor metastasis.

Objective To evaluate the effects of electroacupuncture (EA) at the Baihui (DU 20) and Dazhui (Bill) points on brain derived neurotrophic factor (BDNF) around the area of cerebral infarction and evaluate the relation between learning and memory ability and BDNF. Methods Forty-eight male adult Wistar rats were divided randomly and equally into EA and control groups. The EA group was sub-divided into 1 week, 2 weeks and 3weeks sub-groups. EA was started 24 h after establishing a model of ischemic brain injury and continued for one, two or three weeks. The control group was reared conventionally and was not given any treatment. Morris' water maze test was used to evaluate the rats' learning and memory ability. The expression of BDNF in the CA3 region of the hippo campus was detected using immunohistochemical techniques. Results Learning and memory in the EA groups were better than in the control group, and spatial probe ability was also significantly better. Positive expression of BDNF was detected in the hippocampal CA3 region of the EA group rats, and it was significantly greater than that in the control group. Conclusion Learning and memory after cerebral infarction can be affected by EA at the Baihui and Dazhui points. The effect might be related with increased BDNF expression in the hippocampal CA3 region.

Objective To investigate the role of local aldosterone system in oncofetal fibronectin ( oncofetal FN) mRNA expression and reactive oxygen species (ROS) production in human mesangial cells (HMCs) exposed to short-term intermittent high glucose and the effect of Fenofibrate.Methods The HMCs were divided into 8 groups:normal glucose(NG) ;osmotic fluctuation(OF) ;mean glucose load (MGL) ;stable high glucose (SHG),short-term intermittent high glucose (IHG) ; intermittent high glucose plus eplerenone (IHGE) ; intermittent high glucose plus fenofibrate(IHGF) ; and normal glucose plus fenofibrate (NGF) groups.The mRNA expression levels of Aldosterone synthase ( CYP11 B2 ),11 β-hydroxysteroid dehydrogenase type 2 ( 11βHSD2 ) and oncofetal FN were determined by RT-PCR.The expression of CYP11B2 protein was determined by western-blot.Aldosterone level in cell culture supernatant was detected by radioimmunoassay.The expression and translocation of mineralocorticoid receptor (MR)protein were assayed with confocal laser scanning microscopy. ROS levels were determined by Fluorescence microscopy and fluorescence microplate reader.Results ( 1 ) MGL,SHG,and IHG groups showed a 2.41,3.63,and 4.45 times increase in CYP11B2 mRNA expression,and a 1.83,2.15,and 2.78 times increase in CYP11B2 protein expression,respectively,compared with NG group (P ＜ 0.05 ).The aldosterone levels of HMCs culture supernatant in MGL,SHG,and IHG groups were also increased,being 1.49,2.04,and 2.54 times of that in NG group ( P＜0.05 ),and the degree of elevation in IHG group was more marked than that in SHG group( P＜0.05 ).MR was activated and translocated from cytosol to nucleus in MGL,SHG,and IHG groups.Quantitative analysis showed the ratioes of cytosol/nucleus fluorescence intensity in MGL,SHG,and IHG groups were 15％,38％,and 53％ decreased as compared with that in NG group,and the decrease was more marked in IHG group ( P＜0.05 ).(2) Oncofetal FN mRNA expression and ROS levels in MGL,SHG,and IHG groups were increased,being 1.54,2.31,3.65 and 1.26,1.91,2.48 times of those in NG group,respectively ( P＜0.05 ),and this increase was more marked in group IHG ( P＜O.05 ).Compared with IHG group,oncofetal FN mRNA expression and ROS levels in group IHGE were significantly decreased by 54％ and 53％,and in group IHGF by 45％ and 39％. ( 3 ) CYP11B2 mRNA,protein,and aldosterone levels in IHGF group were decreased by 74％,59％,and 50％,and the activation of MR in group IHGF was inhibited when the ratio of cytosol/nuclear fluorescence intensity was increased 1.88 fold as compared with that in group IHG ( P＜0.05 ).Conclusions Increased expressions of oncofetal FN and ROS by HMCs induced by short-term intermittent high glucose were nore marked than those induced by stable high glucose.The mechanism was associated with activation of local aldosterone system.Fenofibrate may inhibit the activation of local aldosterone system and alleviate the injury to HMCs induced by intermittent high glucose.

Objective To compare the roles of Toll-like receptor 4 (TLR4)/NF-κB signal pathway in acute lung injury (ALl) induced by blunt chest trauma and by blunt chest trauma-hemorrhagic shock and resuscitation (double hits) in rats.Methods Forty male Sprague-Dawley rats,aged 8 weeks,weighing 240-280 g,were randomly assigned into 3 equal groups (n =10 each) using a random number table:sham operation group (S group),blunt chest trauma group (T group),and blunt chest trauma and hemorrhagic shock and resuscitation group (group THSR).Lung contusion was induced in anesthetized rats by dropping a 300 g weight onto a precordial protective shield to direct the impact force away from the heart and toward the lungs.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min,followed by resuscitation.At 6 h after the model was established,the arterial blood samples were collected for blood gas analysis and detection of tumor necrosis factor-alpha (TNF-α) concentrations in serum.Oxygenation index (PaO2/FiO2) was calculated.The rats were then sacrificed and pulmonary specimens were obtained for determination of TLR4 expression and NF-κB ac tivity (by immunohistochemistry and Western blot) in lung tissues and for microscopic examination.Results Compared with group S,PaO2 and PaO2/FiO2 were significantly decreased,PaCO2 and TNF-α concentrations in serum were increased,TLR4 expression was up-regulated,and NF-κB activity was enhanced in T and THSR groups (P ＜ 0.05).Compared with group T,PaO2 and PaO2/FiO2 were significantly decreased,PaCO2 and TNF-α concentrations in serum were increased,TLR4 expression was up-regulated,and NF-κcB activity was enhanced in THSR group (P ＜ 0.05).The histopathological damage to lung tissues was aggravated in THSR group as compared with T group.Conclusion The role of TLR-4/NF-κB signal pathway in ALI induced by blunt chest traumahemorrhagic shock and resuscitation (double hits) is significantly stronger than that in ALI induced by blunt chest trauma alone in rats.