Japan,one of the countries near China,has obtained great success in medical education through a long time exploration.Professor of R.Harsha Rao from America concluded the drawbacks in the early medical education in Japan,then proposed corresponding blueprint including reforming education systems,breaking the limitation in subjects and establishing comprehensive education system.Japanese medical education has been well promoted,the experience can be used in our country's medical education reforms.

Objective To investigate the character 、difference and clinical meaning of distortion product otoacoustic emissions(DPOAE) and auditory brainstem respons(ABR) in high risk neonates with.Methods DPOAE and ABR were recorded with amplaid MK 22 and GSI60 in 200(400ears) high risk neonates, then compared the results.Results Of 200(400ears) newborns, the presence of DPOAE was 64%(256/400 ears), the presence of ABR was 88.25%(353/400 ears). The co-negativity and co-positivity of DPOAE and ABR were 61.50%(246/400 ears)and 9.25%(37/400 ears). The false negative and false positive of DPOAE were 3.91%(10/256 ears)and 74.31%(107/144 ears).Conclusion The DPOAE test is simple,quick,harmless,sensitive and objective. Because it only reflects the function of the cochlea and has high false positive, we can't make the diagnosis of hearing loss immediately when newborns don't pass the DPOAE test.The ABR test is a reliable method for neonatal hearing screening. The combination of DPOAE and ABR in high risk neonatal hearing screening could increase the accuracy and reliability of the test and retest results.

Objective To investigate the dilating effect of dexmedetomidine (DEX)on isolated vascular smooth muscles and to explore the mechanism.Methods Tension of isolated mesenteric arterial rings of male Sprague-Dawley rats was recorded.The effects of DEX on the rings and the effects of DEX on vascular reaction induced by various drugs were recorded. Results DEX completely relaxed the contraction induced by phenylephrine (PE)and KCl in a concentration-dependent manner in endothelium intact mesenteric arterial rings in rats.The vasodilating effect of DEX was increased by sodium nitroprusside.In phenylephrine (10-5 mol/L)based on pre-vasoconstriction,adding acetylcholine could not suppress DEX’s vasodilating effect.Vasodilation was not related to the endothelial cells.In physiological saline solution without calcium,DEX significantly inhibited the contraction induced by addition of CaCl2 .Conclusion DEX can induce vasodilation in a concentration-dependent manner,which is not dependent on the endothelial cells.

OBJECTIVE To prepare pneumolysin(Pn)by genetic engineering and thereby establish the basis for the study of vaccines against otitis media. METHODS A pair of primers including two restriction sites was designed based on the pneumolysin gene sequence reported by Walker in 1987. The pneumolysin gene was PCR-amplified from pneumococcal DNA. The resulting fragment, digested by restriction enzymes, was ligated into the vector PET-28a and then transformed into host cell E.coli JM109(DE3). RESULTS The sequence of the inserted pneumolysin gene was confirmed by DNA sequencing. CONCLUSION The pneumolysin gene was successfully cloned into the host cell.

BACKGROUND: There are fewer reports about estrogen effects on bone marrow stromal cells (BMSCs). OBJECTIVE: To study the effect of diethylstilbestrol on the osteogenic differentiation of rabbit BMSCs. METHODS: Rabbit BMSCs cultured in vitro were intervened with 0, 10-7, 10-6, 10-5 mol/L diethylstilbestrol, and BMSCs cultured with dexamethasone 10-8 mol/L, β-sodium glycerophosphate 10 mmol/L, and vitamin C 50 mg/L were used as positive controls. RESULTS AND CONCLUSION: 10-6 mol/L diethylstilbestrol significantly improved the proliferative ability of BMSCs at 24, 48, and 72 hours after intervention (P < 0.01). 10-5 mol/L diethylstilbestrol significantly inhibited the proliferation of BMSCs at 48 hours after intervention as well as 10-7 mol/L diethylstilbestrol at 72 hours (P < 0.01). Mineralized nodular structures formed at 25 days after intervention with 10-7 mol/L diethylstilbestrol. Alkaline phosphatase activities were remarkably increased at 14 and 21 days after intervention with 10-7, 10-6 mol/L diethylstilbestrol. It has been proved that diethylstilbestrol has an enhancing effect on the osteogenic differentiation of rabbit BMSCs.

Objective:To investigate the expression of Smad1 in gastric carcinoma and the influence on the migration ability of gastric cancer cells.Methods:Collected the protein from the gastric cancer tissues and corresponding adjacent tissues,the expression level of Smad1 was detected by Western blot.In HGC-27 gastric cancer cells as the research object,the carrier cells transfected with overexpression of Smad1 (p-EGFP-C1/Smad1) and Smad1 small interfering RNA (Smad1 siRNA),while transfection of p-EGFP-C1 and siRNA control as control.MTT to detect cell proliferation.Cell migration ability was detected with cell scratch test.The expression levels of MMP-9,MMP-2,p-Akt and Akt in ceils were detected by Western blot.Akt signal pathway inhibitor LY294002 (20 μg/ml) in gastric adenocarcinoma ceils,MTT for cell proliferation,cell scratch assay for cell migratior.The expression levels of MMP-9,MMP-2,p-Akt,Akt were detected by Western blot.Results:Smadl in gastric carcinoma was significantly lower than the adjacent tissues (P＜ 0.01).The cell survival rate and migration rate of p-EGFP-C1/Smad1 group were significantly lower than that of p-EGFP-C1 group (P＜0.01).The cell survival rate and migration rate of Smad1 siRNA group were significantly higher than those in the siRNA control group (P＜0.01).The expression levels of Akt protein in P-EGFP-C1,p-EGFP-C1/Smad1,Smad1 siRNA,siRNA control cells did not change.The expression levels of MMP-9,MMP-2 and p-Akt in p-EGFP-C1/Smad1 group were significantly lower than that in p-EGFP-C1 group (P＜0.01).The expression levels of MMP-9,MMP-2 and p-Akt in Smad1 siRNA group were significantly higher than that in control siRNA (P＜0.01).The cell proliferation and migration trends in gastric cancer cells effected by Akt signaling pathway inhibitor consistent with the p-EGFP-C1/Smadl group.Conclusion:Low expression of Smad1 in gastric cancer tissue.Smad1 can inhibit the proliferation and migration of gastric cancer cells,the mechanism of action is related to the Akt signaling pathway.

Objective To investigate the different radioprotective effects of total flavonoids of Astragalus (TFA) on human normal mesenchymal stem cells(hMSCs) and hepatoma cells injured by 60 Coγ-ray radiation.Methods hMSCs and HepG-2 cells were cultured and randomly divided into TFA-treated and untreated groups.The cells of different groups were irradiated with 60 Co γ-rays at the dose of 6 Gy.MTT method was utilized to detect the survival rates of the hMSCs and HepG-2 cells pretreated or untreated with TFA before irradiation.Cell clone formation test was used to measure the cellular radiosensitivity.The apoptosis rates of different groups were determined by flow cytometer assay.The expression rates of the apoptosis-promoting proteins Fas and Bax and the apoptosis-inhibiting protein Bcl-2 were analyzed by Western blotting.Results MTT showed that the survival rates of hMSCs pretreated by TFA were 1.15-1.95 times higher than that of the pure irradiation group.On the contrary,the survival rates of the TFA pretreated HepG-2 cells were only 0.53-0.23 times that of the pure irradiation group.There was a good dose-effect relationship between the cell survival rate and the TFA concentration.Cell clone formation rate indicated that combined treatment of TFA and radiation inhibited the cell proliferation more effectively than single TFA or pure radiation.Flow cytometry showed that 6,24 and,48 h post-irradiation to 6 Gy,the apoptosis rates of the hMSCs were 23.3% ,11.2% ,and 2.9% ,respectively in the TFA pretreated group and were 29.3% ,24.9% ,and 13.6% in the pure radiation group.However,the apoptosis rates of the HepG-2 cells at 6,24,and 48 h post-irradiation to 6 Gy were 11.6% ,17.3% ,and 20.1% ,respectively in the TFA pretreated group and were 6.9% ,9.3% ,and 15.8% ,respectively in the direct radiation group.Western blotting showed that the expression levels of Fas and Bax proteins in the HepG-2 cells were significantly higher in the TFA pretreated group than in the pure radiation group.On the contrary,the expression level of the apoptosis inhibiting protein Bcl-2 was significantly lower in the TFA pretreated group than in the pure radiation group.Conclusions TFA has obvious effects of radiological protection on human hMSCs and has no effects of radiological protection but effects of apoptosis enhancement on hepatoma cells.The promotion of apoptosis of TFA on hepatoma cells is primarily through increasing the expression of apoptotic proteins such as Fas and Bax and reducing the expression of anti-apoptotic protein Bcl-2.

Freshwater crabs (Sinopotamon denticulatum) were examined for metacercariae. Cats and dogs were also examined for Paragonimus infection. Questionnairing was carried out on health knowledge and behaviors among local residents in a village of Baokang County, Hubei Province. Results showed that the infection rate of Paragonimus skrjabini metacercariae in Sinopotamon denticulatum was 20.5% (46/214), with 15.6% (20/128) in a mining area and 30.2%(26/86) for the non-mining area respectively ( ?2=6.5, P0.05). Questionnairing showed that dogs and cats were with the habit of foraging and defecating at streams and children had the habits of eating raw or under-cooked crabs. The natural and ecological environments are in favor of the life cycle of P. skrjabini.

Objective The aim of this study was to screen and verify the proteins interacting with Vimentin,investigate the regulatory relationship between FABP5 and candidate proteins,and further explore the mechanism of FABP5 in hepatocellular carcinoma.Methods Immunoprecipitation combined with tandem mass spectrometry(IP-MS)was used to screen the proteins that bind to FABP5.The binding relationship between FABP5 and candi-date interacting proteins was verified from the exogenous and endogenous levels by Co-immune precipitation assay(Co-IP).RT-qPCR,Western blot and immunofluorescence were used to observe the effect of knockdown FABP5 on the transcription and translation of Vimentin in HCC cells.The effect of overexpressing FABP5 on the cytoskeleton of HCC cell was observed by phalloidin staining.Results 336 potential target proteins that bind to FABP5 were identi-fied through IP-MS.Based on literature,five candidate proteins related to tumors were selected,namely PRDX1,PRSS3,PKM,HSP90AA1,and Vimentin.The binding relationship between FABP5 and Vimentin protein was con-firmed through both exogenous and endogenous Co-IP.Knockdown FABP5 has no significant effect on the expression of Vimentin mRNA,but it can inhibit the expression of Vimentin protein,and overexpression of FABP5 can affect the cytoskeleton of HCC cell.Conclusions FABP5 promotes the migration and invasion of HCC cells by the regula-tion of Vimentin and the influence of cytoskeletal remodeling,and thus it is expected to be a potential target for anti-HCC and provide new ideas for the treatment of HCC.

Objective To investigate the use of endoscopic retrograde cholangiopancreatography(ERCP)combined with laparoscopic cholecystectomy(LC)one-stage approach in the treatment of gallstones complicated with calculus of common bile duct in a hybrid operating room.Methods 21 patients with gallstones complicated with calculus of common bile duct were selected to undergoing ERCP bile duct stone removal under total intravenous anesthesia or combined intravenous and inhalation anesthesia,followed by LC.Results 18 successfully completed the one-stage surgery of ERCP combined with LC,3 were changed to laparoscopic common bile duct exploration(LCBDE),there was no laparotomy,8 cases were complicated by hyperamylemia after surgery,and there were no serious complications such as pancreatitis,gastrointestinal bleeding and perforation.Conclusion It is safe and feasible to use ERCP combined with LC one-stage approach to treat gallstones complicated with calculus of common bile duct in the hybrid operating room,which simplifies the surgical process.