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.

China ; Occupational Health Services ; methods ; organization & administration

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.

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.

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.

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

OBJECTIVETo evaluate the effect of length of tibial stump on proprioceptive recovery after anterior cruciate ligament (ACL) reconstruction.

METHODSFrom March 2011 to December 2011,42 patients with ACL tear were performed with reconstruction. The patients were divided randomly into three groups: group A, the patients with remained length of tibial stump ranging from 15 to 20 mm, including 8 males and 6 females, with an average age of (28.2 ± 6.6) years old; group B, the patients with remained length of tibial stump ranging from 5 to 10 mm, including 9 males and 5 females, with an average age of (27.9 ± 6.4) years old; group C, the patients with remained length of tibial stump less than 2 mm, including 9 males and 5 females, with an average age of (28.6 ± 6.8) years, old. The stability of knee were assessed by Lanchman test and anterior drawer test. The function of knee was assessed by Lysholm score and Tegner activity rating. The proprioceptive recovery was evaluated by assessing the passive reproduction of the angles with limb movement into flexion and extension in no weight bearing position.

RESULTSLanchman test and anterior drawer test of all patients were negative at 6 months postoperatively., and there was one case of positive outcome in each group at 12 month and 18 month postoperatively. Lysholm score and Tegner activity score of all patients at 18 month postoperatively were significantly better than that preoperatively, and there were no significant difference among three groups at 18 month postoperatively. There were no significant difference in the production of the angles at flexion to 20° and extention to 80° and 20° among these groups before and after operation. The reproduction of the angles of group A and B at flexion to 50° and extention to 50° at 6 month postoperatively were significantly better than that of group A and B preoperatively and that of group C at 6 month postoperatively, and there were no significant difference among three groups at 18 month postoperatively. The reproduction of the angles of group A and B at flexion to 80° at 12 month postoperatively were significantly better than that of group A and B preoperatively and that of group C at 12 month postoperatively, and there were no significant difference among three groups at 18 month postoperatively.

CONCLUSIONPreservation of tibial stump in ACL reconstruction has benefit in proprioceptive recovery at early stage postoperatively and the length of tibial stump should be reserved with a range from 5 to 10 mm.

Adult ; Anterior Cruciate Ligament ; physiopathology ; surgery ; Anterior Cruciate Ligament Injuries ; Anterior Cruciate Ligament Reconstruction ; Female ; Humans ; Knee Joint ; physiopathology ; surgery ; Male ; Proprioception ; Range of Motion, Articular ; Tibia ; chemistry ; physiopathology ; surgery ; Young Adult