Objective To observe the pituitary tumor transforming gene (PTTG), estradiol-stimulated protein 1 (ESP1) expressions in S, G 2/M phases in A549, SPC-A-1 cell lines. Methods The mammalian cells for cell cycle was synchronizated by thymidine, the total RNA in the S, G 2/M phases was collected, and the total RNA was observed by Real-time PCR. Results PTTG expression in A549 and SPC-A-1 cell lines was higher in S phase than that in G 2/M phase while no PTTG expression in MRC-5 cells in S, G 2/M phases. ESP1 expression in A549, SPC-A-1, MRC-5 cell lines was lower in S phase than that in G 2/M phase. Conclusion That no PTTG in S and G 2/M phases in the normal cell line and PTTG expression in the lung cancer cell line higher in S phase than that in G 2/M phase suggested that the aneuploidy may be caused by the overexpression of PTTG. ESP1 expression was lower in S phase than G 2/M phase in all of the three cell lines, suggesting that sister chromatids separation depend on the active ESP1.

There were a lot of differences between the American and Chinese medical education.Students in American should obtain bachelor's degree of humanities or science before reciveing medical doctor's degree at graduation.High altitude militray medicine is a characteristic discipline of the third military medical university,aming at training general doctors for the plateau.During the designing of high altitude militray medicine curriculum and its teaching process,we actively learned from the advanced experiences of American based on the reality,optimizd currulum,improved teaching methods,greatly cultivated the students' protential in order to increase the qualify of high altitude militray medicine personnel.

Aim To explore the effect of nNOS specific inhibitor 7-nitrioindazole(7-NI) on the neurogenesis after focal cerebral ischemia in the adult mice dentate gyrus.Methods Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO).A bromodeoxyuridine(BrdU) method was used to identify the proliferated cells in hippocampal dentate gyrus.Step-down test was performed to assay the learning and memory function.Results The number of BrdU~+ cells was increased in the ipsilateral but not contralateral dentate gyrus after focal cerebral ischemia.7-NI promoted the increment in the ipsilateral dentate gyrus significantly.7-NI also improved the survival of the new-born cells,as well as the learning and memory function of ischemic mice.Conclusion Our results indicate that nNOS down-regulates the neurogenesis after focal cerebral ischemia.

OBJECTIVE:To improve the synthesis technology of sorafenib tosylate (Ⅰ). METHODS:The intermediate N-methyl-(4-chlorpyridin-2-yl)carboxamide(Ⅳ)was obtained by chlorination and amidation with 2-picolinic acid as the starting com-pound. Meanwhile,N-[4-chloro-3-(triflouromethyl)phenyl]-N′-(4-hydroxyphenyl)urea (Ⅶ) was prepared from aminophenol and 4-chloro-3-trifluoromethyl phenyl isocyanate,which was obtained from 4-chloro-3-(trifluoromethyl)aniline(Ⅴ)by reaction with initiator triethylamine. Sorafenib tosylate was synthesized from Ⅳ and Ⅶ with potassium tert-butoxide by condensation and salt for-mation. The target compound was characterized by 1H-NMR. RESULTS:The target compound was confirmed as Ⅰ,with the over-all yield of 74% [based on 4-chloro-3-(trifluoromethylanilin)]. The purification of chromatogram was 94%. Optimized technology improves yield and simplifies multi-step intermediate decompression,distillation and purification process;the application of initia-tor shortens the duration of reaction. CONCLUSIONS:Ⅰ is prepared successfully,and raw material can be obtained easily and is easy to operate with high yield.

ObjectiveTo explore the clinical effects of forearm cutaneous neurovascular flaps to be used on repairing remote wound.Methods17 cases of soft tissue defect of head and facial region and upper extremity were repaired with forearm cutaneous neurovascular flaps.Results The flaps of 16 cases were entirely survived, partial necrosis appeared in 1 case and healed after changing dressing.Following up for 3 to 6 months, the quality and shape of all the flaps and function recovery of all patients were satisfied.ConclusionsWith adequate blood supply, the survival rate of forearm cutaneous neurovascular flaps is high and the procedure to repair remote wound is easy, and it is one of best ways to repair the soft tissue defect in head and facial region and upper extremity with forearm cutaneous neurovascular flaps.

AIM: To study the protective effects of HIF-1α gene transfection on hypoxic injury in human HepG2 cells. METHODS: After gene transfection, HepG2 cells were randomly divided into 4 groups: normoxia with Ad-GFP transfected group, normoxia with Ad-HIF-1 transfected group, hypoxia with Ad-GFP transfected group and hypoxia with Ad-HIF-1 transfected group. LDH leaking rate, cell viability, contents of NO and ROS, the iNOS activity were measured. RESULTS: High levels of HIF-1α mRNA and protein were detected in Ad-HIF transfected HepG2 cells. Cell viability was significantly lower in Ad-GFP transfected-hypoxia group than that in Ad-GFP transfected-normoxia group (P<0.05). No marked difference of cell viability was found between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group. ROS was significantly higher in Ad-GFP transfected-hypoxia group than that in Ad-GFP transfected-normoxia group (P<0.05), while no marked difference was found either between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group or between Ad-HIF transfected-hypoxia group and Ad-GFP transfected-hypoxia group. The content of NO and iNOS activity were significantly higher in Ad-HIF transfected-normoxia group and Ad-GFP transfected-hypoxia group than those in Ad-GFP transfected-normoxia group (P<0.05), no marked difference was found either between Ad-HIF transfected-hypoxia group and Ad-GFP transfected-hypoxia group or between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group. CONCLUSION: Higher HIF-1α expression is contributed to protective effects against hypoxic injury in HepG2 cells, the mechanisms of which may be correlated with promoting expression of gene regulated by HIF-1 and restraining over-expression of injure factors.

Objective To investigate the different effects of internal and external biliary drainage on the levels of TNF-α,NO in blood serum and NO,inducible nitric oxide synthase(iNOS) in gastric mucosa with obstructive jaundice and explore the mechanism of gastric mucosa damage and compare the quality of internal and external drainage.Methods To establish the animal model:one hundred male adult Sprague-Dawley rats were randomly assigned to five groups:obstructive jaundice for 7 days group (OJ-7 d group),obstructive jaundice for 14 days (OJ-14 d group),internal biliary drainage group(ID group),external drainage group(ED group)and sham operation group(SH group).The OJ-7 group rats were executed 7 days after the first operation,the rest four groups were executed 7 days after the secondary surgery,leaving serum and gastric mucosa.Tested the levels of INF-α,NO in blood serum and NO,iNOS in gastric mucosa and did pathological biopsy of gastric mucosa.Results The levels of TNF-α,NO in blood serum and NO,iNOS in gastric mucosa significantly increased in OJ-7 group.The levels of TNF-α,NO in blood serum and NO,iNOS in gastric mucosa continued to rise in OJ-14 group,and was significantly higher than OJ-7 group,the gastric mucosa was obviously impaired.The levels of TNF-α,Internal drainage can significantly NO in blood serum and NO,iNOS in gastric mucosa were significantly lower in ID group,gastric mucosal damage significantly reduced.Conclusions After internal drainage,iNOS expression was inhited in gastric mucosal,NO production was significantly reduced,led to protect the gastric mucosa,which provide experimental basis for choice in OJ clinical treatment.

Objective To investigate effect of sodium cyanide(NaCN)at different concentrations on the mitochondrial respiratory oxygen consumption,the mitochondrial membrane potential(MMP)and activity of the complex Ⅳ of mitochondrial breath train in in vitro liver mitochondria from the rats exposed to simulated high altitude hypoxia,and to explore the characteristics of energy metabolism in the mitochondria from the rats subjected to cyanide poisoning during acute hypoxia exposure.Methods Adult Sprague-Dawley(SD)rats were set randomly into control and acute hypoxia groups(n=8 in each group).The acute hypoxic rats were exposed to simulate 5 000 m high altitude in a hypobaric chamber 23 h/d for 3 d.Rats in the control group were bred in the normoxia condition at the same time.The liver mitochondria were isolated by centrifugation.Mitochondrial oxidative respiratory consumption and activity of the complex Ⅳ was measured by Clark electrode after the treatment of NaCN at 0,0.01,0.1 and 0.25 mmol/L respectively,so as to calculate mitochondrial state 3 respiration(ST3),state 4 respiration(ST4),respiratory control rate(RCR),the rate of oxidative phosphorylation(OPR),and oxygen consumption rate of the complex Ⅳ.MMP was detected by Rhodamine 123 method at the above-mentioned concentrations of NaCN.Results NaCNat0.01,0.1and0.25mmol/Linhibited the mito-chondrial oxidative respiratory function,and decreased MMP significantly.The inhibitory effects of NaCN onenergy metabolism in mitochondria was in a dose-dependent manner.Compared with the treatment of NaCN atthe corresponding concentration in the control group,mitochondrial function in the acute hypoxia group was in-hibited more seriously.Conclusion Acute hypoxia exacerbates the inhibitory effects of sodium cyanide on en-ergy metabolism in rat liver mitochondria.Its mechanism might be relevant to the decoupling of oxidative phos-phorylation,functional down-regulation of complexⅣin respiratory chain and changes of the mitochondrialmembrane potential in liver mitochondria from rats exposed to acute hypoxia.

Objective To explore the expression and relationship of PTEN and mTOR in the development of cholangiocarcinoma,determine the effect of the PI3K/PTEN/AKT/mTOR signal transduction on the(development) of cholangiocarcinoma.Methods The expression of mTOR and PTEN in the cholangiocarcinoma was detected by the immunohistochemistry and RT-PCR method.Results Compared to normal tissues,the(expression) of mTOR gene in cholangiocarcinoma significantly increased,but the expression of PTEN gene(decreased).There was a negative correlation between mTOR gene and PTEN gene expression in(cholangiocarcinoma). Conclusions The expression of mTOR gene in cholangiocarcinoma was increased and the expression of PTEN was decreased.It suggested that the mTOR gene and PTEN gene could play an important role in the process of development of cholangiocarcinoma.

This study aimed at exploring the application of microtox technology to the evaluation of comprehensive toxicity of Sheng Mai injections.Characteristic parameters of vibrio fischeri (CS234) were measured by methodology inspection under different conditions to optimize the reaction condition with reliable technology.It was the first experiment to accomplish the comprehensive toxicity of Sheng Mai injections from various manufacturers using vibrio fischeri under the target condition.It was found that parameters of the optimum condition contained 0.9 mL recovery liquid volume in each freeze-dried vial and 50 μ L bacteria suspension of each sample,being included in 2 mL solution in aggregate under 10 mins' detection time after adding bacteria suspension with the favorable pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million within 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15％.Under this optimum detection condition,it was found that the EC50 values were 22.10％,34.10％ and 46.04％,respectively,presenting significant statistical differences (P＜0.05).These results demonstrated that the growth of biological detection standards of Sheng Mai injection was highlighted a long way to go.Besides,the microtox-based fast test for the evaluation of the comprehensive toxicity of Sheng Mai injection showed a prospective application to controlling the quality of different products from manufacturers.