To improve the quality of the pediatric education in clinical clerkship and to bring us more in line with international practice,we applied the PBL bedside teaching in the clini-cal clerkship of the pediatric education.Compared with the traditional bedside teaching,better re-sults could be obtained with improvement in the consciousness of study,communication skill,the combination of basic and clinical knowledge,and the establishment of the clinical thought and evi-dence-based thought.

BACKGROUND:Fibrin glue is a natural biodegradable scaffold, which can be used for tissue-engineered scaffolds, and is increasingly used as seed cel carrier for tissue engineering repair. OBJECTIVE:To investigate the biocompatibility in vitro of rabbit fascia fibroblasts and fibrin glue. METHODS:Tissue explants adherent method was used to culture fibroblasts from subcutaneous deep fascia tissue of New Zealand white rabbits. The fibroblasts could be passaged with trypsin digestion method. Suspension of passage four fibroblasts was co-cultured with fibrin glue. Morphology and proliferation of fibroblasts on the surface of fibrin glue were dynamical y observed under the inverted phase contrast microscope. At 5 days after co-culture, fibroblasts were identified by immunofluorescence staining under the laser scanning confocal microscope. The fibroblast growth and adhesion were observed under the scanning electron microscope. RESULTS AND CONCLUSION:There was no significant difference in fibroblast morphology between co-culture fibroblasts and pure culture fibroblasts with inverted phase contrast microscope. Scanning electron microscope demonstrated that fibroblasts ful y extended in fibrin glue surface, and showed a good adhesion between the“pseudopodium”and fibrin glue, and secreted matrix material. It is clear that the fibrin glue did not alter the morphologic features of fibroblasts. Laser scanning confocal microscope revealed that fibroblasts were positive for vimentin. These verified that properties of fibroblasts did not change after they were seeded in fibrin glue surface and did not be induced to differentiate. There is a very good biocompatibility between fascia fibroblasts and fibrin glue in vitro.

China ; Occupational Health Services ; methods ; organization & administration

AIM: To evaluate the effects and safety of intravitreal injection of triamcinolone acetonide ( TA ) or conbercept combined with macular laser grid photocoagulation in the treatment of macular edema secondary to retinal vein occlusion( RVO) . ·METHODS: Fifty cases ( 50 eyes ) with macular edema secondary to retinal vein occlusion were selected and assigned to 2 groups: intravitreal injection of TA or conbercept, and laser photocoagulation after 7d. Best corrected visual acuity ( BCVA ) , fundus examination, optical coherence tomography ( OCT ) and intraocular pressure ( IOP ) were examined before intravitreous injection and 14d, 1 and 3mo after laser, fundus fluorescein angiography(FFA) were examined 3mo after treatment. The postoperative results at each time point were compared with preoperative values. · RESULTS: Two kinds of treatment compared with preoperative, the BCVA all increased in various degrees. At 14d after intravitreous injection, 1 and 3mo after laser, the ratio of vision improved in TA group was 76%, 80%, 68%, conbercept group was 88%, 92%, 88%, BCVA of two groups in each period all had varying degrees of increase than preoperative. The best BCVA acquired at 1mo after treatment. The macular thickness after treatment was significantly lower than preoperative in two groups. At preoperative, 14d, 1 and 3mo after treatment, the macular thickness in TA group was 557. 5 ± 150. 9,301. 7±120. 1, 262. 7 ± 131. 2, 338. 1 ± 146. 5μm; the macular thickness in conbercept group was 569. 4 ± 135. 9, 282. 3 ± 133. 5, 259. 5 ± 116. 4, 307. 8 ± 122. 6μm. The macular thickness of the two groups were significantly different between preoperative and postoperative. · CONCLUSION: The combination of intravitreous injection of TA or conbercept with macular laser grid photocoagulation can be an effective method in the treatment of macular edema secondary to RVO, conbercept treatment is more effective and security.

Objective: To modify EBP(endotoxin binding peptide), clone and express the mutate of EBP gene and gain purified mEBP.Method: mEBPgene was cloned by PCR site-directed mutagenesis. PinpointXa-3/mEBP expression vector was designed to express human mEBP as a fusion protein in BL21 (DE3) pLysS. Digested engineering bacteria by lysozyme and collected inclusion bodies.Fusion protein was purified by Pinpoint TM Xa purification system and cleaved by factorXa,mEBP was purified by RP-HPLC. Results: Mutations at residues 5 and 18(Gln→Lys) was obtained by PCR site-directed mutagenesis, expressed and purified mEBP successfully.Conclusions: Obtaining of purified mEBP lay a foundation for its biological activity research.

The L-cysteine desulfhydrase gene (cd) of Pseudomonas sp.TS1138 was amplified by PCR,and the amplified gene was recombined in the cloning vector pBluescript SKII.The 1.2kb DNA fragment containing cd was sequenced,and its homology with other desulfhydrases was blast; then the cd was cloned into the expression vector pET-21a(+), and afterward expressed by IPTG inducement.The expression protein was purified by Ni-NTA His-Bind Resin.Then the expression protein was identified by the method of activity staining of desulfhydrase, and the characterization of L-cysteine desulfhydrase and the critical role it played in the L-cysteine biosynthetic pathway were discussed.

The L-ATC hydrolase and L-SCC amidohydrolase which convert L-ATC to L-cysteine in Pseudomonas sp.TS-1138 are purified about 83.9 and 90.3 fold by salting-out method, Sephadex G-75 gel chromatography, DEAE-cellulose 52 ion exchange and Sephadex G-100 gel chromatography, etc. The purified enzyemes are both demonstrated by SDS-PAGE to be a homogeneous protein. Their molecular weight are about 37.5kD and42.8kDa respectively. The optimum reaction temperature are both 35℃, and the optimum pH are 7.0 and 8.0 respectively. The Km of the two enzymes are 0.67 mmol/L and 0.15 mmol/L, and the Vmax are 0.39?10 -3mmol/L?min and 0.42?10 -3mmol/L?min respectively.

OBJECTIVETo study the therapeutic mechanism of Zhizhu Pill (ZP) for treating functional dyspepsia (FD) rats.

METHODSTotally 30 ten-day-old male rats were randomly divided into the normal control group (n =10) and the model group (n = 20). The FD rat model was induced using gastric administration of 0.1% iodoacetamide (IA) combined tail clamping. The model was evaluated when rats were 8-week old. Successfully modeled rats were randomly divided into the model group (n = 10) and the ZP group (n = 10). Rats in the normal group and the model group were administered with normal saline by gastrogavage, while those in the ZP group were administered with ZP Decoction (2 mL/100 g) by gastrogavage. All medication lasted for 7 successive days. The contractile activity in in vitro longitudinal gastric muscle was recorded using Power Lab biological signal collecting system. The expression of growth hormone secretagogue receptor (GHSR) in stomach of FD rats was detected using Western blot and immunohistochemistry (IHC).

RESULTSCompared with the normal group, average frequencies of gastric contraction and changing rates of amplitude obviously decreased in the model group (P < 0.05). Results of Western blot and IHC showed that the expression of GHSR decreased in the model group (P < 0.01). Compared with the model group, average frequencies of gastric contraction and changing rates of amplitude obviously increased in the ZP group (P < 0.05). Results of Western blot and IHC showed that the expression of GHSR increased in the ZP group (P < 0.01).

CONCLUSIONZP could promote the gastric motility in FD rats induced by gastric administration of IA combined tail clamping, and its mechanism might be related to up-regulating GHSR protein level.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Dyspepsia ; drug therapy ; Gastrointestinal Motility ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; metabolism ; Random Allocation ; Rats ; Receptors, Ghrelin ; metabolism

Communicable Diseases ; epidemiology ; Disease Outbreaks ; Humans ; Population Surveillance ; methods

Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.