We have reformed our patterns and methods of graduation examination of the medical majors. That is the multi-stage comprehensive graduation examination of the clinical interns, which consists of 70% close written test of clinical theories,20% practical test of clinical skills and also oral test of related theories, and 10% writing of the medical documents. It has proved that these measures have improved the interns' attitude toward practice, aroused their initiative and positivity, and led the practitioners to attach more importance to the combination of clinical practice and theories, thus leading to better clinical education.

BACKGROUND:The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cel s, which can form a limit or amplification cascade to realize the complex immune function of T cel s. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk. OBJECTIVE:To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cel s. METHODS:The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cel s. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus). RESULTS AND CONCLUSION:Confocal microscope showed that cel transfection efficiency was more than 95%and Cbp was located on the cel membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cel s shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cel s was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cel s, the high expression of Cbp can decrease the uniformity of cel s and increase the necrosis cel s, thus inhibiting the signal transduction.

Background and purpose: p21-activated kinase 5 (PAK5) is a recently identified member of PAKs that regulate many intracellular processes such as cytoskeleton remodeling, cell proliferation, cell differentiation, gene transcription and cell apoptosis. Recently, studies found that PAK5 was overexpressed in some cancer such as gastric and colon cancer. However, the expression status and biological function of PAK5 in osteosarcoma are not clearly known. The objective of this study was to investigate the expression of PAK5 in osteosarcoma tissue and their relationships with the prognosis of osteosarcoma. Methods: The expression of PAK5 was detected by using immunohistochemical method in 92 specimens of human osteosarcoma tissues and 33 cases of osteoclastoma tissue, respectively. Results: The positive rate of PAK5 was 71.7% (66/92) in all the 92 cases of osteosarcoma. PAK5 expressions were not related to clinical variables such as gender, age, tumor location, tumor size, histological type and local recurrence, but signiifcantly related to Enneking grade, tumor cell necrosis rate and lung metastasis, and the high expression of PAK5 may reduce the efifciency of chemotherapy. Survival analysis indicated that high expression of PAK5 correlated with poor prognosis of patients with osteosarcoma. Univariate survival analysis showed that the signiifcant prognostic factors were tumor size, Enneking grade, local recurrence, lung metastasis and expression levels of PAK5. COX multivariate regression identified that the PAK5 expression levels (P=0.001) and lung metastasis (P=0.015) were independent prognostic factors of patients with osteosarcoma. Conclusion:The positive expressions of PAK5 closely correlate with Enneking grade, tumor cell necrosis rate and lung metastasis. Detection of PAK5 may be used as a molecular marker for prognosis of osteosarcoma. The high expression of PAK5 may reduce the efifciency of chemotherapy.

To study the effects of surfactants on wettability of excipients, the contact angles of six types of surfactants on the surface of two common excipients and mixture of three surfactants with excipients were measured using hypsometry method. The results demonstrated that contact angle of water on the surface of excipients was associated with hydrophilcity of excipients. Contact angle was lowered with increase in hydrophilic groups of excipient molecules. The sequence of contact angle from small to large was starch < sodium benzoate < polyvinylpyrrolidone < sodium carboxymethylcellulose < sodium alginate < chitosan < hydroxypropyl methyl cellulose

Objective:To investigate the function of CD 59 in LAT induced T lymphocytes'proliferation and activation.Methods:Transfected LAT-GFP recombinant lentiviral vectors into Jurkat cells and established a fusion-protein stable express cell line ( Jurkat-GFP ).Junket-GFP cells were transfected with pSUPER-siCD59 plasmids by electroporetion or stimulated by anti-CD59 antibody.The cellular locations of CD 59 and LAT were observed under fluorescence microscope with the immunofluorescence cytochem -istry.The cells proliferation were measured by MTT assay.Furthermore,Western blot was used to detect the total and phosphorylation levels of several down-stream proteins after T cell activated .Results: Jurkat-GFP cells successfully transfected with pSUPER-siCD59 plasmids showed lower fluorescence staining.CD59 and LAT distributed uniformly on the cell surface before stimulated with anti-CD59 antibody and formed clusters once upon stimulation.Jurkat-GFP cells stimulated with anti-CD59 antibody showed a higher level of pro-liferation and protein phosphorylation ,compared with the others.Conclusion:CD59 contributed to LAT induced signaling transduction of T lymphocytes ,and stimulated CD59 molecule partly promoted T cell activation.

Objective To investigate the expression pattern and significance of two importantoocyte-secreted factors:growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)during oocyte maturation in women with polycystic ovary syndrome(PCOS)and infertile women due to husband factors.Methods Total of 25 oocytes[9 at germinal vesicle GV stage,9 at M Ⅰ stage and 7 at M Ⅱ stage]were obtained from 12 patients with PCOS and 82 oocytes(29 at GV stage,26 at M Ⅰ stage and 27 at M Ⅱ stage)were from 56 controls.The nested quantitative real time(RT)PCR was uscd to detect the abundance of GDF9 and BMP15 mRNA in each oocyte.Results(1)The expression level of GDF9 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 44.8(4.2-529.0),27.6(9.8-172.7)and 49.0(0.2-65.9)respectively,the expression in were 149.9(55.4-387.9),29.9(2.5-205.8)and 657.8(149.4-1376.2)in control group,respectively.The expression of GDF9 mRNA at M Ⅱ stage was significantly lower in PCOS group than in controls(P ＜ 0.01),however,the differences didn't reach statistical significant at GV or M Ⅰ stage between the two groups(P ＞ 0.05).The expression of GDF9 mRNA displayed some changes at different maturation stage in controls(P ＜ 0.05,P ＜ 0.01),however,the expression didn't demonstrate any dynamic changes in PCOS group(P ＞ 0.05).(2)The expression level BMP15 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 0.1(0.1-22.0),3.2(0.6-55.0)and 6.4(3.2-8.5),respectively,the expression were 41.6(6.5-96.1),4.0(2.0-10.4)and 49.7(2.3-139.5)in control group,respectively.The expression of BMP15 mRNA at GV stage was significantly lower in PCOS group than in controls(P ＜ 0.01),however,the differences were not significant at M Ⅰ or M Ⅱ stage between the two groups(P ＞0.05).The expression of BMP15 mRNA also displayed some changes at different maturation stage in controls(P ＜ 0.05),however,the level didn't demonstrate any dynamic changes in PCOS group(P ＞ 0.05).Conclusion It was suggested that the low expression of oocyte secreted factors in mature oocytes from PCOS patients might be associated with impaired oocyte quality and developmental competence in PCOS.

Objective To study the effects of dammarane sapogenins ( DS-1226 ) on sleep interruption-induced learning and memory impairment in mice.Methods 130 SPF healthy 5 -6-week old male ICR mice were randomly divided into control, model, DS-1226 low dose, DS-1226 medium dose and DS-1226 high dose groups.The behavioral alterations in open field (OF), Morris water maze (MWM) and step-through (ST) tests were detected at 15 days after rotating drum-induced sleep interruption ( SI) .Results The total distance, movement speed, total duration of movement were increased in OF test ( P<0.05, vs.the model group) after treatment.The latency of place navigation was increased from day 5 in the MWM test after 15 d sleep interruption, and the number of crossing in the target quadrant and the percent distance in target quadrant were decreased after 15 d sleep interruption ( P <0.05, vs.the control group), while the latency of place navigation was decreased, and the percent distance in target quadrant and percent time in target quadrant after high dose DS-1226 oral administration ( P<0.05, vs.the model group) were increased.Error times, distance in dark chamber, time in dark chamber and immobility time in dark chamber were increased in training of step through test ( P<0.05, vs.the control group);while these indexes were decreased after DS-1226 oral administration ( P<0.05, vs.the model group) .But there was no significant difference in the step through testing course.Conclusions The results show that orally administrated DS-1226 can ameliorate SI-induced learning and memory impairment, and there is a significant dosage-effect relationship.

The cDNA of an i type lysozyme was cloned from Stichopus japonicus (named as SjLys). The DNA fragment of the mature SjLys was subcloned into expression vector of pET-32a (+) to construct the recombinant plasmid of pET32a (+)-SjLys. The recombinant plasmid was then transformed into Escherichia coli BL21 (DE3) pLysS and induced by isopropylthio-beta-D-galactoside (IPTG). The recombinant protein expressed as inclusion bodies was denatured, partially purified and refolded to be an active form. The bacteriolytic activity of recombinant protein purified by the metal-chelating was 19.2 U/mg. The antibacterial activity of the purified recombinant SjLys (rSjLys) was analyzed. The rSjLys protein displayed inhibitive effect on the growth of the tested Gram-positive and Gram-negative bacteria. In particular, rSjLys had a strong inhibitive activity on Vibrio parahaemolyticus and Pseudomonas aeruginosa, both the most common pathogenic bacteria in the marine animals. The heat-treated rSjLys exhibited more potent activities against all tested bacteria. These results indicated that the S. japonicus lysozyme was the enzyme with combined enzymatic (glycosidase) and non-enzymatic antibacterial action, and it had a wide antibacterial spectrum. Therefore, it is suggested that the S. japonicus lysozyme should be one of the important molecules against pathogens in the innate immunity of sea cucumbers.
Animals ; Anti-Bacterial Agents ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Muramidase ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Stichopus ; enzymology

Objective To investigate the effects of chronic restraint stress on learning and memory in Wistar and Sprague-Dawley (SD) rats.Methods Healthy adult male Wistar rats (n=6) and Sprague-Dawley rats (n=6) were subjected to restraint stress 10 h daily for 28 days.After that, all rats were tested for recognition memory by novel object recognition test , and spatial memory and working memory by Morris water maze test .Results After restraint for 10 h daily for 28 days, the restraint rats of the two strains demonstrated lower discrimination index (DI)than the control group, but on-ly SD rats showed significant difference ( P<0.05 ) .The restraint SD rats showed higher escape latency than the control rats, and on the 5th day the difference became significant (P<0.05), and there was no significant difference between Wistar restraint and control rats .The working memory test showed that restraint SD rats exibited longer escape latency than the control rats (P<0.05), while Wistar rats didn’t show significant difference between the two groups .Conclusions The results of this study demonstrate that the impairments of learning and memory in SD rats subjected to restraint 10 hour per day for 28 days are more serious than that in the Wistar rats .Therefore , SD rats may be a better choice as an animal model to study the effects of chronic restraint stress on learning and memory impairment .

Objective To analyze the characteristics of adverse events following immunization ( AEFI) with varicella attenuated live vaccine ( VarV) in Hebei province during 2012—2014, to evaluate the safety of VarV and to provide supportive evidences for planning chicken pox prevention and control strate-gies in Hebei province.Methods The AEFI data associated with VarV in Hebei province during 2012 to 2014 were collected from the national AEFI information management system and were analyzed by using the method of descriptive epidemiology.Results A total of 239 cases reported during 2012 to 2014 were col-lected from the national AEFI information management system, with a male to female ratio of 1.44 ∶1.The estimated incidence rate was 25.51 per 100 000 doses.One case was serious AEFI with an incidence rate of 0.11 per 100 000 doses.Most of the AEFI cases were children under 3 years old and identified in scattered inhibiting children and kindergarten kids.The main symptoms of common vaccine reactions were fever, local swelling and induration .The rare vaccine reactions were presented as anaphylactic rashes .Conclusion The reported incidence rate of VarV associated AEFI in Hebei province was less than expected.However, the estimated incidence rate of common vaccine reactions in Hebei province was higher than that showed on the Analysis on Surveillance Data of Post-marketing Immunization Safety for Varicella Attenuated Live Vac-cine in China, 2010—2012.Attentions to the AEFI after immunization with VarV should also be paid in fu-ture.