Buyang huanwu decoction attenuates cerebral ischemia-reperfusion injury in rats by regulating autophagy through AMPK/mTOR/ULKl signaling pathway / 中国药理学通报

Aim To explore the mechanism of Buyang huanwu decoction attenuating cerebral ischemia-reper- fusion injury in rats by regulating autophagv through AMPK/mTOR/ULKl signaling pathway.Methods Left eerebral ischemia model in rats was established by modified thread ocelusion method, then the rats in each group were given medieine onee every 24 hours for 3 times.After 72 hours of reperfusion, the nerve injury and the changes of cerebral infarction volume were observed; the morphology, number and apoptosis of nerve cells were observed by Nissl staining and TUNEL staining; the expression of autophagy protein and AMPK/mTOR/LJLKl autophagy signaling pathway related proteins were detected by Western blot.Results Buyang huanwu decoction could improve the neurological deficit of rats, reduce the volume of cere bral infarction and neuronal apoptosis, reduce the pathological injury of brain tissues, inhibit the phosphorylation activation of AMPK, relieve the inhibition of AMPK on downstream mTOR and LJLK1 , promote the phosphorylation activation of both, and inhibit autophagy.AMPK agonist metformin increased the level of autophagy and reversed the protective effect of Buyang huanwu decoction on cerebral ischemia-reperfusion injury in rats.Conclusion Buyang huanwu decoction mediates AMPK/mTOR/ULKl autophagy signaling pathway to play a neuroprotective effect on cerebral is- chemia-reperfusion injury in rats.

Ischemic preconditioning inhibits over-expression of arginyl-tRNA synthetase gene Rars in ischemia-injured neurons / 华中科技大学学报（医学）（英德文版）

The expression changes of Rars gene in ischemia-injured neurons were investigated by detecting its translational product arginyl-tRNA synthetase (ArgRS), and the inhibitory effects of ischemic preconditioning (IPC) on Rars gene were explored. Both IPC model and prolonged ischemia (PI) model were established by using the classic oxygen glucose deprivation (OGD) method. The primary cultured neurons were assigned into the following groups: the experimental group (IPC+PI group), undergoing PI after a short period of IPC; the conditional control group (PI control group), subjected to PI without IPC; blank control group, the normally cultured neurons. The Rars transcriptional activities and ArgRS expression levels were measured at different time points after re-oxygenation (3 h/6 h/12 h/24 h). Data were collected and statistically analyzed. Compared to the blank control group, the Rars activities and ArgRS levels were significantly increased in PI control group, peaking at the time point of 6 h after re-oxygenation. Rars activities and ArgRS levels were significantly lower in the experimental group than in the PI control group at different time points after re-oxygenation. PI insult can induce an escalating activity of Rars and lead to ArgRS over-expression in primary cultured neurons. IPC can inhibit the increased Rars activity and down-regulate ArgRS expression of ischemia-insulted neurons. This mechanism may confer ischemic tolerance on neurons.

Vacuum sealing drainage combined with discontinuous windowing technique for repairing large area exposed wound of Achilles tendon / 中国骨伤

<p><b>OBJECTIVE</b>To explore clinical effect of vacuum sealing drainage (VSD) combined with discontinuous windowing technique for repairing large area exposed wounds of Achilles tendon.</p><p><b>METHODS</b>From July 2009 to May 2014, 11 patients with large exposed wounds of Achilles tendon were treated, including 5 males and 6 females with an average age of 43 years old (aged from 7 to 65 years old). Among them, 4 cases were skin necrosis caused by heavy objects abrasion and contusion; 3 cases were caused by distal tibiofibula fractures; 3 cases were caused by bicycle-spoke injuries; 1 case was caused by diabetes. Areas of exposed Achilles tendon were from 6 cmx3 cm to 14 cmx5 cm without tendon rupture or bone exposed. After debridement, discontinuous fenestration on Achilles tendon was made by knife blade parallel with longitudinal axis of Achilles tendon, combined with Vacuum Sealing Drainage (VSD) treatment.</p><p><b>RESULTS</b>After drainage treatment with one VSD cycle (5 to 7 days), abundant fresh granulation tissues were growing on all wounds and survived well after the second phase dermatoplasty. All patients were followed up for 12 to 24 months, the color of skin flap was good, the texture was soft without burst. At 3 to 4 months after operation, subcutaneous fat was appeared under the flap, the skin was sliding, movement of ankle joints was good. No delayed Achilles tendon rupture were occurred.</p><p><b>CONCLUSION</b>Vacuum sealing drainage (VSD) combined with discontinuous fenestration is a simple, safe and effective method for repairing large area exposed wounds of Achilles tendon,which could minimize the secondary damage caused by wounds of skin flap grafting.</p>

Clinical observations of emergent PTCA combined with Lipo-PGE_1 for the young patients with acute myocardial infarction / 中国基层医药

Objective To evaluate the clinical effect in the treatment of the young patients(≤45 years old) with acute myocardial infarction(AMI)underwent emergent percutaneous transluminal coronary angioplasty(PTCA) combined with Lipo-PGE_1.Methods 39 patients with AMI(paroxysm within 12 hours),were underwent emergent PTCA(coronary stem performed in some patients),including 18 cases which were treated combined with Lipo-PGE_1 in the mean time.And the clinical efficacy and the results of short-period follow-up were recorded.Results The in- farctive vasculars were re-open in 37 patients(23 cares were routinely placed translunrinal srents),and the successful rate was 94.9 %.Those who also used Lipo-PGE_1 were re-open in 17 patients.The successful rate was 94.4 %,their ST segments on EKG 30 minutes after operations reduced significantly than that of patients who did not use Lipo- PGE_1,their cardial functions were also improved significantly 24 hours after operations and no side effects on blood pressure and heart rate were observed.Conclusion The emergent PTCA combined with Lipo-PGE_1 for acute my- ocardial infarction can protect the cardial function and show a better early therapy effect.

Mutations of tumor suppressor gene PTEN mutations in hepatocellular carcinoma and its implications in tumor proliferation and apoptosis / 中华病理学杂志

<p><b>OBJECTIVE</b>To study mutations of tumor suppressor gene PTEN in human hepatocellular carcinomas and its effects on the proliferation and apoptosis of hepatocellular carcinoma cell line HHCC.</p><p><b>METHODS</b>(1) PCR-SSCP and sequence analysis were used to detect the mutations of the 5th and 8th exon of PTEN in 42 cases of human primary hepatocellular carcinoma. (2) Eukaryotic expression vectors of the wild-type (pEGFP-wt-PTEN) and the mutant type (pEGFP-PTEN, G129R) of PTEN were constructed. Lipofectamine 2000 mediated gene transfection was used to transfect hepatocellular carcinoma cell line HHCC, in which the PTEN protein is not expressed. Culture medium containing G418 was used to select stable transfectants. MTT colorimetry was used to analyze the proliferation ability of selected cell lines. Naive HHCC cells and HHCC cells transfected with empty vector (pEGFP-C1) served as controls. (3) TNF-alpha was used to induce apoptosis of selected cell clones.</p><p><b>RESULTS</b>(1) Point mutation involving the 5th exon of PTEN was detected in 4 of 42 primary hepatocellular carcinomas. (2) Compared with the control groups, the proliferation of hepatocellular carcinoma cells was significantly inhibited by the transfection of wild-type PTEN gene, while the transfection with mutant PTEN construct did not significantly change the proliferation. (3) The apoptosis indices of cells transfected with the wild-type and the mutant PTEN genes were 13.8% and 8.1% respectively. Compared with the control, the apoptosis index of HHCC cell transfected by the wild type PTEN was significantly lower (P < 0.05). There were no significant differences between HHCC cells transfected with mutated PTEN gene and the control (P > 0.05). The expression of internal 473-phosphorylated Akt of HHCC was weak, but was enhanced when the cells treated with TNF-alpha. However, it was down regulated by the wild type PTEN.</p><p><b>CONCLUSIONS</b>(1) First time report that PTEN mutations can be found in 9.5% human primary hepatocellular carcinomas. (2) The expression of the wild-type PTEN can suppress the proliferation of HHCC cells, and such suppression was lost when PTEN gene was mutated. (3) PTEN inhibition of the proliferation and the enhancement of apoptosis of hepatocellular carcinoma cells is likely related to a down-regulation of the TNF-alpha induced activation of protein kinase Akt pathway.</p>

Inhibitory effect of tumor suppressor gene PTEN on hepatocellular carcinoma cell line HHCC proliferation and its mechanisms of action / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of tumor suppressor gene PTEN on proliferation and cell cycle of hepatocellular carcinoma cell line HHCC.</p><p><b>METHODS</b>Firstly, eukaryotic expression vectors of wild type and mutated type of PTEN gene were constructed, named as pEGFP-WT-PTEN and pEGFP-PTEN; G129R, respectively. Lipofectamine 2000 was used to transfect the constructed expression vectors into hepatocellular carcinoma cell line HHCC which was PTEN protein negative. G418 was used to select the cell clones constantly expressing PTEN protein. Flow cytometry was used to assay the cell cycle of HHCC transfected by above mentioned eukaryotic expression vectors and non-transfected cell line HHCC. Intrinsic 473-phosphorylated AKT representing the level of active AKT was assayed by Western blot. The non-transfected HHCC served as control.</p><p><b>RESULTS</b>The proliferation of HHCC constantly expressing PTEN protein was obviously inhibited compared with HHCC cells transfected with mutated PTEN gene and empty vectors, and non-transfected HHCC cells. The number of HHCC cells transfected with wild type PTEN gene at G(1) phase, G(2) phase and S phase was 70.8%, 6.8% and 22.4%, respectively. Compared with control group transfected with empty vector, the number of G(1) phase HHCC cells constantly expressing wild type-PTEN protein was significantly higher than that of control. The number of cells in G(2) and S phase was significantly lower than that of control. However, the number of cells in G(1) phase, G(2) phase and S phase of HHCC transfected with mutant PTEN was 63.2%, 10.1% and 26.7%, respectively. There was no significant difference compared with control group. Western blot result showed that the intrinsic level of 473-phosphorylated AKT of HHCC constantly expressing wild type PTEN protein was down-regulated, and that of HHCC transfected with mutated PTEN gene was equal to that of control.</p><p><b>CONCLUSION</b>Wild type PTEN gene can inhibit the proliferation of hepatocellular carcinoma cells at G(1) phase. The mechanism is possibly related with intrinsic activity of AKT, which is down-regulated by wild type PTEN.</p>