Construction of acid-sensitive potassium channel-3 eukaryotic expression plasmid and its express in SH-SY5Y cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To construct the acid-sensitive potassium hannel-3(TASK3) eukaryotic expression plasmid and to establish a stable SH-SY5Y cell line expressing enhanced green fluorescent protein (EGFP)-tagged TASK3.</p><p><b>METHODS</b>TASK3 coding region was subcloned into pEGFP-N1 plasmid to construct a recombinant vector alled pEGFP-TASK3. The correct recombinant expressing plasmid was transfected with X-feet transfection reagent to SH-SY5Y cells. The cell line stably expressiing EGFP tagged-TASK3 gene was established by screening with antibiotic G418 and fluorescence microscope. The expression and localization of the EGFP tagged-TASK3 fusion protein was detected by Western blot and confocal microscope. Exposure of the SH-SY5Y cell line expressing stably TASK3-eGFP fusion proteins was exposed to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability was assessed with cell counting Kit-8 (CCK-8).</p><p><b>RESULTS</b>All the results of identification by PCR, digestion with restriction endonuclease and sequencing indicated that the recombinant eukaryotic expression plasmid pEGFP-TASK3 was constructed correctly. The stable SH-SY5Y cell line expressing EGFP tagged-TASK3 fusion protein was successfully established. Exposure of the wild type SH-SY5Y cells and the stable SH-SY5Y-GFP tag-TASK3 cell line to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability of two group cells significantly reduced with pH declining, and the difference was statistically significant (P < 0.05). Compared with wild type SH-SY5Y cells, the cell viability of stable SH-SYSY-GFP tag-TASK3 cell line increased significantly with the same pH media, and the difference was statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>The eukaryotic expression vector pEGFP-TASK3 is successfully constructed and the cell line stably expressing TASK3-eGFP fusion is established which is important for their fundamental research and potential applications.</p>

Neuroprotective effect of progesterone on focal cerebral ischemia/reperfusion injury in rats and its mechanism / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the neurological protective effects of progesterone (PROG) on focal cerebral ischemia/reperfusion injury in rats and to explore its possible mechanism.</p><p><b>METHODS</b>One handred and twenty male SD rats were divided into three groups randomly: sham-operated group, middle cerebral artery occlusion ( MCAO ) group and PROG + MCAO group( n = 40). The right temporary MCAO model was established by the line-embolism method. The PROG + MCAO group rats were according to 8 mg/kg intraperitoneal injection PROG, after that 30 min, the rats were suffered ischemia/reperfusion. After rats were suffered ischemia for 2 h and reperfusion 0, 24, 48, 72 h stress, the nervous functional defect degree were evaluated by longe scoring, and the expression of two-pore domain K channel 3 (TASK3) mRNA in brain tissue were detected by the real-time PCR.</p><p><b>RESULTS</b>PROG (8 mg/kg) could significantly reduced the nervous functional defect degree in rats after ischemia/reperfusion 24, 48, 72 h (P < 0.05). The results of real-time PCR showed that the TASK3 mRNA expression in the brain tissue at all time points significantly decreased in MCAO group compared with sham-operated group (P < 0.05). However, compared with MCAO group, the expression of TASK3 mRNA in brain tissue at all time points dramatically increased in PROG + MCAO group (P < 0.05).</p><p><b>CONCLUSION</b>PROG can improve the nervous functional defect degree after focal cerebral ischemia/reperfusion injury in rats, and the mechanism might be associated with up-regulating the expression of TASK3 mRNA in brain tissue.</p>

Effects of taurine on the ultrastructure and P2X7 receptor expression in brain following traumatic brain injury in rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effects of taurine on the ultrastructure and P2X7 receptor protein expression in brain following traumatic brain injury (TBI) in rats.</p><p><b>METHODS</b>Forty male SD rats, were divided randomly into four groups that were sham-operated group, TBI group, TBI plus low-dose taurine group and TBI plus high-dose taurine group. The TBI model was established by Marmarou's method, the expression of P2X7 receptor protein in parietal cortex and hippocampus was detected by the immunohistochemical method, the ultrastructure of parietal cortex were observed by transmission electron microscope.</p><p><b>RESULTS</b>Compared with sham-operated group, the positive expression cells of P2X7 receptor protein in parietal cortex and hippocampus of TBI group were significantly increased (P < 0.01). Compared with TBI group, the positive expression cells of P2X7 receptor protein in parietal cortex and hippocampus of TBI plus low-dose taurine group and TBI plus high-dose taurine group were significantly decreased (P <0.01 or P <0.05). Compared with TBI plus low-dose taurine group, the positive expression cells of P2X7 receptor protein in parietal cortex and hippocampus of TBI plus high-dose taurine group were significantly decreased (P < 0.05 or P < 0.01). The pathological damage of parietal cortex in the TBI plus high-dose taurine group was obviously lightened.</p><p><b>CONCLUSION</b>Taurine exerts the neuroprotective effect on TBI in rats, the protective mechanism might be associated with down-regulating the expression of P2X7 receptor protein in parietal cortex and hippocampus.</p>

Influence of Nuclear Factor-?B on Cardiac Myocyte Apoptosis in Septic Shock Mouse / 实用儿科临床杂志

Objective To explore the effect of nuclear factor-kappaB(NF-?B) on cardiac myocyte apoptosis and understand the molecular mechanism of decrease of heart function in septic shock rats.Methods Twenty Wistar rats were randomly divided into the septic shock group and the normal control group.The septic shock group(n=10) were fixed with anaesthesia and made into 1.5 cm slices just along the abdomen midline.Then the roots of appendices were returned to the belly cavity after being ligated and punctured 4 times with the No.14 pinhead.After that,the slices were sewn up layer by layer.After 12 hours,the rats were all sacrificed,the blood taken and the serum separated.In the end,the heart specimens of the septic shock group were set aside.Meanwhile,the normal control group were dealt with in the same way except that they were not subjected to cecal ligation and puncture.Then NF-?B protein levels in cardiac tissue and the index of cardiac myocyte apoptosis were measured.Results NF-?B protein levels in the septic shock group(9 cases were strong masculine gender,1 case was middle masculine gender)elevated significantly compared with the normal control group(8 cases were negative gender,2 cases were weak masculine gender) in cardiac tissue(P