Objective To explore the effects of urinastatin(UTI) on microcirculation of extra pancreatic organs in rats with acute necrotizing pancreatitis(ANP). Methods A total of 48 rats were randomized into control group, ANP group and UTI group. The model of ANP was established by uniform injection of 5% sodium taurocholate solution under pancreatic capsule, only injection of normal saline in control group. Then the rats of UTI group were injected with UTI through the femoral vein, the rats of ANP group and control group were injected with normal saline. The blood flow of lung, kidney and distal small intestine was measured by radioactive biomicrosphere technique at 2 h and 6 h after ANP.Results Compared with the control group, the blood flow of lung, kidney and intestine was decreased significantly in the ANP group at the 2 h and 6 h after ANP ( P

OBJECTIVE:To provide reference for improving teaching effects of Medicinal Chemistry Experiment,and cultivat-ing practical talents that in line with needs of the pharmaceutical industry. METHODS:Based on the training target of medicinal preparation major in our college,the basic idea of establishing Medicinal Chemistry Experiment system was discussed,and teach-ing target,content,process and experimental evaluation results of Medicinal Chemistry Experiment were analyzed in detail. RE-SULTS:The experiment teaching system was established,with teaching target of improving knowledge,ability and quality,teach-ing contents of basic experiment,comprehensive experimental design experiment and open experiment,center of developing practi-cal teaching from before class,in class and after class,and means of diversified performance evaluation. CONCLUSIONS:The es-tablished experiment teaching system is suitable for the teaching of Medicinal Chemistry Experiment in medicinal preparation ma-jor,it has created a new situation that professional,innovation and quality education organic integrate in teaching activity,mobi-lized the students’learning enthusiasm and initiative,exercised the students' professional knowledge and practical ability,cultivat-ed students' creative ability and comprehensive quality and enhanced the effect of experiment teaching.

Objective To investigate the effects of application of pseudoreplica technique in electron microscope observation of A?.Methods With the modification of classical pseudoreplica technique,ADDLs were spotted and concentrated on the gel before negative staining.Results The structures of 5nm to10nm spherical granules and 20 to 30nm coils were visualized by transmission electron microscopy(TEM).Conclusion Pseudoreplica is rapid and effective in ADDLs condensation,negative staining especially in further ultra miero structure observation and research.

OBJECTIVE:To study the dynamic changes of TNF-?/IL-10in model rats with acute necrotizing pancreatitis (ANP)and to study the intervention outcome of salvia miltiorrhiza.METHODS:A total of96rats were randomly divided into three groups(each with32rats):pancreatitis(P)group,salvia miltiorrhiza treatment(T)group and control group(C),the blood samples of rats in each group were taken to determine levels of TNF-?and IL-10and the ratio changes of TNF-?/IL-10.Meanwhile,pancreas tissue sample was collected for pathological scoring.RESULTS:Group P had significantly higher serum levels of TNF-?and IL-10than did group C(P

OBJECTIVE:To evaluate the quality of Bombyx batryticatus medicinal materials in the market by multi-index,and provide reference for clinical rational drug use. METHODS:Referring to the Chinese Pharmacopoeia (2015 edition,part 1),the appearance characters of samples were observed,moisture and the total ash were detected,Fourier transform infrared spectroscopy was adopted to identify the medicinal materials,UV-Visible spectrophotometry and potassium permanganate titration were respec-tively used to determine the contents of active ingredient protein and ammonium oxalate. RESULTS:The moisture ranged in 2.84%-4.07%(RSD<5%,n=3),total ash in 7.79%-34.32%(RSD<5%,n=3),UV and FTIR spectra were basically similar, and both can reflect the characteristic peaks of protein peptides. Protein content ranged in 29.87%-57.19%(RSD<0.5%,n=3), ammonium oxalate content in 0.27%-4.77%(RSD<5%,n=3). CONCLUSIONS:The 10 batches of samples are authentic and processed products,moisture fits the requirement in pharmacopoeia while total ash does not;and the main ingredient is protein pep-tides. However,effective protein polypeptide content shows great differences as well as ammonium oxalate.

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection

Objective To evaluate the prophylactic efficacy of compound sulfamethoxazole (SMZco)combined with ganciclovir on severe pulmonary infection in the early stage of renal transplantation. Methods Between January 2005 and January 2006,two hundred and forty renal allograft patients in our hospital were enrolled in this study.All the patients were divided into two groups.Group A(n=84)received oral SMZco combined with intravenous ganciclovir.Group B(n=156)received intravenous ganciclovir only as control.According to the time of SMZco administration,group A was divided into two subgroups:group A1(within 2weeks after transplantation,n=43)and group A2(more than 2 weeks after transplantation,n=41).All the patients were followed up for 9 months.Incidence of pulmonary infection and effects on graft function by SMZco at different time point were investigated. Results The incidence of severe pulmonary infection and mortality of infection were significantly lower in group A than those in group B (2/84 vs 16/156,P=0.027;0/2 vs 2/16,P＜0.01).There were no significant differences between two groups in terms of age,gender,warm or cold ischemia,complement dependent cytotoxieity test results,incidence of urinary infection and Scr.The incidence of elevatedScr was significantly lower in group A2 than that in group A1(15/43 vs 2/41,P＜0.01),however,all the elevated Scr returned to basal level within 1 week after SMZco was discontinued.Conclusions Oral SMZco combined with ganciclovir administration after renal transplantation is effective on preventing severe pulmonary infection and thus improves graft and recipient survival.The administration of oral SMZco initiated more than 2 weeks after transplantation is better for graft function.

Objective To study the effects of elodronate-liposome on inducing apoptosis of alve-olar macrophages from rats with acute neetotizing pancreatitis (ANP).Methods The AMs of eight rats with ANP were isolated, purified then incubated from broehoalveolar lavage by the differing rates of attachment of the various cell types in a forty-well cell culture plate.Then they were randomized in-to five groups including control group,blank liposome group( 50 μ1, 100 μ1),clodronate-liposome group (50μ1,100μ1).Values of OD were determined by MTT.AO fluorescence and haematoxylin dye were employed to determine the apoptosis of the AMs.Results There were no significant differences be-tween control group and blank liposome group(50 μ1, 100 μ1).Significant differences were found be-tween control group and clodronate-liposome group(50 μ1, 100 μ1).There were no marked differences between blank liposome group(50μ1, 100 μ1)and clodronate-liposome group(50 μ1,100μ1).AO fluo-rescence and haematoxylin dye were available to define the apoptosis of the AMs.Conclusion Clodr-onate-liposome can effectively induce the apoptosis of the AMs.

Objective To investigate the tissue remodeling and cell alignment of TDBM scaffolds seeded with rabbit tenocytes under the cycle dynamic tensile force or static tension-free culture in vitro. Methods TDBM were made by ourselves, and uniaxial cyclic tendon stretching device was designed and manufactured on our own. Primary tenocytes were isolated from the Achilles tendon of three-day-old New Zealand white rabbits and seeded into scaffolds, and were cultured collectively in DMEM in vitro. Samples were divided into two groups:dynamic tension-loaded group, and static tension-free group. Fresh natural tendons were used to be positive control. The experiment's time was six weeks. The scaffold-cell complexes were harvested at 3 and 7 days of culture for Inverted microscope and scanning electron micrograph (SEM) analysis. The morphological characters of the samples, including the general view, HE and Masson's dyeing, were observed at 2, 4 and 6 weeks. In addition, the gene expression of the I-type collagen and III-type collagen of the samples was detected by using Real time PCR at every week. Set fresh natural tendon as control. Results The inverted microscope and SEM showed that it was nice compatible condition between the tendon cells and TD-BM scaffold. In addition, the tendon of tension-loaded group revealed a structure of longitudinally aligned collagen fi bers and dense structure of collagen fibers arranged in orderly form a unique corrugated structure. Tenocytes layer located between the col-lagen fibers and aligned longitudinally along the force axis, with increased matrix deposition after the 3th week showed by RT-PCR. The cell/matrix ratio decreases. When cultured to 6 weeks, the tissue structure was very similar to that of fresh natural ten-don pattern. By contrast, HE and Masson's staining revealed the collagen fibro-tissue structure in tension-free groups with disorga-nized matrix structure and randomly distributed cells. Collagen fibers were gradually degraded with time. In tension-loaded group, Real-time PCR showed that gene expression of I-type collagen and III-type collagen gradually increase, but in tension-free group, the relative gene expression of I-type collagen and III-type collagen was highest at 3rd week, and from that time the relative expres-sion gradually decrease. Conclusion Under the dynamic stress, the TDBM scaffolds seeded with rabbit tenocytes can promote extra-cellular matrix biosynthesis and tendon structure remodeling, which is a promising method for tendon tissue engineering.

Objective To express and characterize the virus-like particles( VLPs) of H5 subtype containing of hemagglutinin ( HA ) and matrix 1 ( M1 ) protein by using Baculovirus-insect cells .Methods Full length genes encoding HA protein from the A/Indonesia/05/2005(H5N1) strain and the M1 protein from the A/Anhui/01/2005 ( H5N1 ) strain were cloned into a baculovirus expression vector to construct pFBD-M1-HA.The expression of HA and M1 proteins were detected by Western blot and indirect immunoflu-orescence after the transfection of Spodoptra frugiperda (Sf9) insect cells with recombinant baculovirus.Pu-rified VLPs were analyzed by SDS-PAGE and visualized with transmission electron microscope.The biologi-cal activity of purified VLPs was detected by hemagglutination test.Results The HA and M1 proteins of H5 subtype expressed by baculovirus-insect cells could be self-assembled into the functional mature VLPs.The hemagglutination titer of VLPs was as high as 1024 HAU/50μl.Conclusion The H5 subtype VLPs as pre-pared in this study would pave a way for the development of a candidate recombinant A ( H5) vaccine.