Aim To investigate the drug-containing serum of Rhizoma drynariae on osteogenesis differentiation of bone marrow mesenchymal stem, and discuss the possible mechanism.Methods BMSCs were cultured in media with different concentrations of medicine containing serum.BMSCs proliferation ability was detected in 3,5,7,9 days by CCK-8.ALP activity was detected after 7,10,14 days′ induction.After 3 weeks culturing, alizarin red staining was performed to observe the formation of calcium nodules.The expression of β-catenin,LRP5,RUNX-2 and Osteriex mRNA were detected using RT-PCR.The protein expression of β-catenin,LRP5 was detected using Elisa method.Results Rhizoma drynariae drug-containing serum could obviously promote the proliferation of BMSCs and calci-fied nodule formation.Besides, the ALP activity was improved in a certain period of time.The expression of β-catenin,LRP5,GSK-3β,RUNX-2 and Osteriex mRNA were significantly up-regulated,and the protein expression of β-catenin,LRP5 was up-regulated too.The expression of GSK-3β was down-trgulated.Conclusions Rhizoma drynariae drug-containing serum promotes mineralization and osteogenic differentiationof BMSCs, and the mechanism is closely related with activating WNT/beta-catenin signaling pathway, raising the beta-catenin, LRP5, RUNX-2, and Osteriex mRNA expression, beta-catenin, LRP5 protein expression,and down-regulation of GSK-3β mRNA expression.

Objective To study the diagnosis,prophylaxis and treatment of splenic artery steal syndrome (SASS),and to evaluate their clinical outcomes in recipients who underwent orthotopic liver transplantation (OLT).Methods 1 385 consecutive patients who suffered from liver cirrhosis and had undergone OLT in our hospital between Jan,2004 and Dec,2013 were studied.We hypothesized that patients were at risk of SASS when the calibre of the splenic artery (SA) was 1.5 times larger than the common hepatic artery (CHA) together with splenomegaly (318 patients,23.0％).Further surveillance with Doppler ultrasound (DUS) was carried out immediately at CHA reperfusion during operation.When a sluggish peak systolic velocity (PSV) ＜ 30 cm/s or no flow was detected in a patent hepatic artery,prophylactic SA banding (SAB) was considered.127 patients (39.9％) who fulfilled these criteria were recruited to the intervention group to undergo SAB.Eventually,patients who developed SASS were treated with coil-embolization of the SA (SAE),re-anastomosis of the HA to aorta (HTA),ligation of SA (SAL) or splenectomy (SPT),or retransplantation.Results SAB resulted in immediately increase in the mean PSV of the HA from 19.3 ±5.5 cm/s to 45.9 ± 9.1 cm/s (P ＜ 0.05),and resistance index (RI) of the HA rehabilitated to reasonable levels (0.5 ～0.8),without any HA or biliary related complication in all the 127 patients.17 patients in the control group were identified to have SASS (8.9％).5 of these 17 patients required emergency treatment by coil-embolization.Of the remaining 12 patients,11 who developed hepatic artery thrombosis secondary to SASS required to undergo embolectomy or thrombolysis followed by HTA (4 patients),SAL (3 patients),SPT (5 patients).Three of these patients finally required re-OLT.All these patients obtained acceptable results by these salvage strategies,except 2 out of the 12 patients who died from liver failure.Conclusions SASS is an important but it is often and under-diagnosed cause of graft ischemia after OLT.Prophylactic SAB should be introduced to patients at risk of developing SASS in order to obtain satisfactory results.Coil-embolization of SA shortly after diagnosis is an effective salvage intervention to prevent further progression to develop devastating consequences.

To prepare recombinant human retinol binding protein 4 (RBP4) by using the baculovirus expression system and to detect its immunogenicity, the fusion DNA fragment of secretory signal peptide SS64 and human RBP4 gene was subcloned into a baculovirus transfer vector pFastBac-dual(pFBd), and the corresponding recombinant transfer plasmid was transformed into E. coli strain DH10bac, after transposition recombinant shuttle bacmid was screened out. The logarithmic phase Sf9 cells were transfected with the recombinant bacmid and then the recombinant baculovirus containing hRBP4 expression box were generated. After amplification of recombinant baculovirus, the recombinant baculovirus seeds were obtained. To express human RBP4, logarithmic phase Sf9 cells were infected with the virus seeds and SDS-PAGE and Western blotting were used to detect and identify the expression. Finally, to prepare a batch of RBP4 protein, logarithmic phase Sf9 cells in suspension culture were infected with recombinant baculovirus seeds and the supernatant was harvested after 120 hours post-infection for purification. Finally for preparation of polyclonal antibody and evaluation of immunogenicity, the recombinant hRBP4 from insect cells and from E. coli were immunized rabbits. Restriction enzyme digestion and sequencing confirmed that the recombinant baculovirus transfer plasmid was constructed correctly, and subsequently recombinant RBP4-bacmid was generated successfully. SDS-PAGE and Western blotting analysis suggested that human RBP4 protein was highly expressed in Sf9 cells with the molecular weight of approximately 23 kDa. The recombinant RBP4 protein could be secreted into the medium efficiently, and the expression level was calculated amount of 100 mg/L. Finally the rabbit antiserum was harvested after recombinant RBP4 immunization, therein the titer of antiserum against baculovirus recombinant RBP4 is 1:100 000 whereas the titer of antiserum against E. coli recombinant RBP4 is only 1:10 000. Overall, human RBP4 was high efficiently expressed successfully with good antigenicity in baculovirus system, and high affinity antiserum was obtained. A solid foundation was laid for the next step of the preparation of human serum RBP4 detection kit.
Animals ; Baculoviridae ; genetics ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Humans ; Immune Sera ; Insecta ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Retinol-Binding Proteins, Plasma ; biosynthesis ; immunology ; Sf9 Cells ; metabolism ; Transfection

OBJECTIVE：To provide re ference f or hospital pharmacy prevention and control management during novel coronavirus（SARS-CoV-2）infection epidemic period. METHODS ：Based on 5M1E analysis method ，according to the needs of epidemic prevention and control ，it is necessary to analyze the risks of 5 aspects as personnel ，equipment and materials ，methods， environment，monitoring of the pharmacy work in hospital ，and establish the prevention and control strategy of hospital pharmacy infection in response to the epidemic situation of novel coronavirus pneumonia （COVID-19）according to the corresponding risks. RESULTS & CONCLUSIONS ：Personnel management strategies include carrying out pharmacist prevention and control training ， focusing on physical and mental health of pharmacists during infection prevention and control ；equipment and materials management strategies include strengthening equipment disinfection management and strengthening the management of materials for infection prevention and control ；method management strategies include developing emergency plans for infection prevention and control，standardizing individual infection prevention and control method ；environment management strategies include environment cleaning and disinfection management ，infection exposure management of related medical material ，medical waste management ； monitoring management strategies include strengthening pharmacists infection monitoring and evaluating pharmacists ’prevention and control effect. By establishing the strategy for COVID- 19 epidemic prevention and control ，it can effectively guiding pharmacists to carry out epidemic prevention and control.

OBJECTIVE：To provid e reference for pharmaceutical workers to better understand Novel Coronavirus Infection ： Expert Consensus on Guidance and Prevention Strategies for Hospital Pharmacists and the Pharmacy Workforce （hereinafter referred to as “expert consensus ”），and to apply and practice in specific work ，so as to give full play to the role of pharmacists to help fight the epidemic.METHODS ：The background of the formulation and revision of the expert consensus were introduced ，and its main contents and viewpoints were interpreted. RESULTS & CONCLUSIONS ：The text of expert consensus is divided into 8 parts，mainly including disease diagnosis and treatment [SARS-CoV- 2 infection related background ，clinical manifestations and diagnosis， treatment]，hospital pharmacy （prevention and control strategy ，work guidance ），drug and facility support management（key drug/facility/equipment support ，management and use of the drug in special circumstances ），information sources and related resources ，etc.，which comprehensively and detailedly provide information ，guidance and strategies for coronavirus SARS-CoV-2 infection prevention and control to play the role of pharmacists in hospital pharmacy well ，do well in the protection of staff in different pharmaceutical posts ，drug security work in response to epidemic situation ，and develop pharmaceutical care. So far，the understanding of SARS-CoV- 2 in the pharmaceutical industry is relatively limited. Based on the accumulated experience and progress in epidemic prevention and control ，the expert consensus will be updated and improved continuously ，so as to provide guidance and help for hospital pharmaceutical personnel.