Intrathecal infection of TRESK overexpression adenovirus to relieve neuropathic pain via inhibiting JNK activation and neuronal apoptosis / 实用医学杂志

Objective To observe the effect of intrathecal injection of TRESK overexpression adenoviruson phosphorylation of JNK and apoptosis of neurons in neuropathic pain rats.Methods Seventy-two male SD rats were randomly divided into six groups:groups C,S,NP,T,V,and NS,12 for each group.SNI was administrated to rats in groups NP,T,V and NS.TRESK adenovirus and negative virus were intrathecally injected after use of SNI in groups T and V,while equal volume of NS was injected to rats in group NS.MWT and TWL were measured at 1 day before operation(baseline,BL)and at 1,3,7 and 14 days after operation (days 1,3,7,and 14).Six rats in each group were sacrificed at D7 to determinate the expression of TRESK protein of DRG.The other rats were sacrificed at D14 to determinate neural apoptosis and the expressions of caspase3 and p-JNK of DRG.Results As compared with groups C,S and T,the expression of TRESK protein was significantly decreased at D7 in groups NP,NS and V (P＜0.05).Compared with groups C and S,MWT was significantly decreased at days 1,3,7 and 14 (P＜0.05),phosphorylation of JNK in DRG was significantly increased at D14 (P＜0.05),neuronal apoptosis rate and expressions of Caspase3 of DRG were significantly increased at D14 (P＜0.05) in groups NP,T,NS and V.Compared with groups NP,V and NS,MWT was significantly increased at time points of days 1,3,7 and 14 in group T (P＜0.05),phosphorylation of JNK of in DRG was significantly decreased at D14 in group T (P＜0.05),neuronal apoptosis rate and expression of Caspase3 of DRG were significantly decreased at D14 in group T (P＜0.05).Intrathecal injection ofpAd/CMV/VS-DEST-TRESK obviously reduced mechanical hyperalgesia,upregulated TRESK expression,and lowered JNK phosphorylation and NP in SNI rat.Conclusions Intrathecal injection of TRESK over expression adenovirus relieves NP via inhibiting JNK activation and neuronal apoptosis.

Changes in APPL1 expression during renal fibrosis in a mouse model of renal ischemia-reperfusion injury / 中华麻醉学杂志

Objective To evaluate the changes in the expression of adaptor protein containing pleck-strin homobgy domain, phosphotyrosine-binding domain and a leucine zipper motif 1(APPL1)during renal fibrosis in a mouse model of renal ischemia-reperfusion(I∕R)injury. Methods Twenty-four male C57BL∕6 mice, aged 8 weeks, weighing 20-25 g, were divided into 2 groups(n=12 each)using a random number table: sham operation group(S group)and renal I∕R group.The model of renal I∕R injury was established by clipping the bilateral renal pedicles for 30 min followed by reperfusion in group I∕R.Six mice were selected at 2 days of reperfusion, and venous blood samples were collected for determination of serum concentrations of blood urea nitrogen and creatinine.The animals were then sacrificed, the renal specimens were obtained for microscopic examination of tubular necrosis with a light microscope, and the damage to the renal tubules was scored using a semi-quantitative method.Six mice were sacrificed at 14 days of reperfusion, and the renal specimens were obtained for assessment of the degree of renal fibrosis(using picric acid-sirius red staining) and for determination of the expression of collagen type 1, fibronectin and α-smooth muscle actin in renal tis-sues(by Western blot or immunofluorescence method). At 2 and 14 days of reperfusion, the expression of APPL1 in renal tissues was detected by Western blot and the expression of APPL1 mRNA in renal tissues by real-time polymerase chain reaction. Results Compared with group S, the serum concentrations of blood u-rea nitrogen and creatinine, scores of renal tubular damage and degree of renal fibrosis were significantly in-creased at 2 days of reperfusion, the expression of collagen type 1, fibronectin and α-smooth muscle actin in renal tissues was up-regulated at 14 days of reperfusion, and the expression of APPL1 protein and mRNA was up-regulated at 2 and 14 days of reperfusion in group I∕R(P<0.05). Conclusion Up-regulated expression of APPL1 may be involved in the process of renal fibrosis in a mouse model of renal I∕R injury.

The Effect of Media Reports of Violence Injury Medical Events on Medical Students / 中国医学伦理学

Objectives:To understand the medical students'cognition of violence injury medical and the impact of medical injury events reports on medical students.Methods:A sample survey was conducted among 480 medical students in a medical university using self-developed questionnaire.Results:A total of 63.67％ of the medical students get violence injury events information through television news;more than 90％ of medical students were concerned about the media reports of violence injury medical events.The media reports of violence injury medical events had an important impact on medical students,and especially television reports had the greatest impact.41.2％ of the students would enhance their awareness of protection;37.9％ of medical students thought that work passion and enthusiasm would decline;the impact of reports on men was bigger than women (P ＜ 0.05).Conclusion:The report of violence injury medical events has an important impact on medical students.The media should improve self-discipline,create a harmonious public opinion environment and promote the healthy development of doctor-patient relationship.

AMPK attenuates inflammation to reduce fibrosis induced by acute ischemia reperfusion injury in mice / 中华肾脏病杂志

Objective To observe the effect of adenosine monophosphate activated protein kinase (AMPK) on attenuating inflammation in fibrosis induced hy acute ischemia reperfusion injury (IRI) in mice.Methods Forty eight male C57BL/6 mice were randomly divided into four groups:sham operation group (sham group),IRI group,AMPK inhibitor+IRI group (AMPK/IRI group) and normal saline+IRI group (NS/IRI group),12 mice each group.The mice with renal IRI were occluded for 30 min through clipping bilateral renal pedicle,then released renal perfusion.Mice in sham group were performed the separation of renal pedicle without clipping.Mice in AMPK/IRI group and NS/IRI group were respectively intraperitoneal injected AMPK inhibitor and normal saline before IRI.At the 2 d after operation,6 randomly-selected mice from each group were blooded by extraction eyeball to detect BUN and Scr.The renal histopathological changes were observed through HE staining.The mRNA expression of IL-1β,IL-6 and TNF-α was detected by real time PCR,and the level of AMPK phosphorylation was detected by Western blotting.At the 14 d after operation,Collagen 1 (COL1),α-SMA and fibronectin (FN) were detected by immunofluorescence and Western blotting in 6 remained mice from each group.The degree of kidney fibrosis was observed through sirus red staining.Results Compared with those in sham group,tubular interstitial damage was aggravated (P ＜ 0.05),BUN and Scr were increased (P ＜ 0.05),the mRNA expression of IL-1β,IL-6 and TNF-α was increased at the 2 d after operation (all P ＜ 0.05),and the level of AMPK phosphorylation was activated in IRI group and NS/IRI group (all P ＜ 0.05);the degree of kidney fibrosis and the expression of COL1,α-SMA and FN were increased obviously at the 14 d (all P ＜ 0.05).Compared with those in IRI group,in AMPK/IRI group tubular interstitial damage was aggravated (P ＜ 0.05),BUN and Scr were increased (all P ＜ 0.05),the mRNA expression of IL-1β,IL-6 and TNF-α was increased at the 2 d (all P ＜ 0.05),and the level of AMPK phosphorylation was decreased (P ＜ 0.05).Moreover,the degree of kidney fibrosis and the expression of COLI,α-SMA and FN were increased obviously at the 14 d in AMPK/IRI group (all P ＜0.05).Conclusions AMPK can ameliorate the acute renal ischemia reperfusion injury induce fibrosis in mice,and the mechanism may be related to the decrease of inflammatory reaction.

Curcumin inhibits HeLa cell invasion and migration by decreasing inducible nitric oxide synthase / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effects of curcumin against HeLa cell invasion and migration and explore the underlying mechanisms.</p><p><b>METHODS</b>HeLa cells were exposed to curcumin treatment at the concentrations of 0, 10, 25, 50, 100, 150 and 200 µmol/L for 24 h. MTT and TUNEL assays were used to assess the cell proliferation inhibition and apoptosis, respectively. Transwell assay was used to evaluate the invasiveness and migration of the treated cells, and RT-PCR and Western blotting were employed to detect the changes in the expression of inducible nitric oxide synthase (iNOS), and MMP-9 and E-cad, the 2 markers of cell invasion and migration, were detected by Western blotting. The capacity of NO production in HeLa cells was measured by Griess method.</p><p><b>RESULTS</b>Curcumin inhibited the proliferation of HeLa cells by inducing cell apoptosis in a concentration-dependent manner. Curcumin inhibited the invasion and migration of HeLa cells by increasing E-cad expression and decreasing MMP-9 expression, and also decreased the expression level of iNOS and NO production in the cells.</p><p><b>CONCLUSION</b>Curcumin inhibits the invasion and migration of HeLa cells by decreasing the expression of iNOS.</p>

Changes in TRESK mRNA expression in dorsal root ganglion in a rat model of neuropathic pain / 中华麻醉学杂志

Objective To evaluate the changes in the expression of diplopore potassium ion channel TRESK mRNA in dorsal root ganlion (DRG) in rats with neuropathic pain (NP) .Methods Thirty-two male SD rats weighing 220-250 g were randomly divided into 2 groups ( n = 16 each) : group sham operation (group S) and group NP. NP was induced by ligation and severance of left tibial and common fibular nerves according to the technique described by Decosterd. Eight rats in each group were sacrificed 1 day before and 14 day after operation and their L4,5 DRGs in the operated side were isolated for determination of TRESK mRNA expression by RT-PCR. In the remaining 8 rats in each group paw withdrawal threshold to mechanical stimuli ( MWT) and paw withdrawal latency to a thermal nociceptive stimulus (TWL) were measured at 1 day before (baseline) and 1, 3, 5, 7, 14 day after operation. Results MWT was significantly lower in group NP than in group S. The TRESK mRNA expression in L4,5 DRGs in the operated side was significantly decreased after operation as compared with the baseline before operation in group NP and was significantly lower in group NP than in group S. Conclusion The development and maintenance of NP may be closely related with down-regulation of TRESK mRNA.

Contraction of rat TRESK gene recombinant adenovirus vector / 中华麻醉学杂志

Objective To construct rat TRESK gene recombinant adenovirus expression vector.Methods TRESK full-length cDNA was cloned from rat dorsal root ganglion cells and confirmed by RT-PCR and sequencing. pAd/CMV/V5-DEST-TRESK was then constructed under the control of CMV-promotor. DH5α colibacillus was translated and the positive recombinants were subsequently identified by PCR and DNA sequencing. 293T cells were cotransfected and packed to produce adenovirus. Results The titer of virus was tested using Hole-by-dilution titer method. The full length of TRESK from rat dorsal root ganglion cells is 781 bp. It was demonstrated that the DNA sequencing was completely consistent with TRESK sequencing of rat recorded in GeneBank. The PCR amplification of the pAd/CMV/V5-DEST-TRESK gDNA was matched with pAD-GFP blank vector as anticipated. The titer of the concentrated virus was 1.31×109 TU/ml. Conclusion Rat TRESK gene recombinant adenovirus vector is constructed successfully.

Effect of dexmedetomidine on brachial plexus block with ropivacaine and upper extremity ischemia-reperfusion injury in patients undergoing upper extremity surgery / 中华麻醉学杂志

Objective To investigate the effect of dexmedetomidine on brachial plexus block with ropivacaine and upper extremity ischemia-reperfusion (I/R) injury in patients undergoing upper extremity surgery. Methods Forty ASA Ⅰ or Ⅱ patients of both sexes, aged 18-55 yr, weighing 45-80 kg, scheduled forupper extremity surgery under brachial plexus block, were randomly divided into 2 groups ( n = 20 each): control group ( group C )and dexmedtomidine group (group D). In group C, brachial plexus block was performed using 0.5% ropivacaine 30 ml. In group D, brachial plexus block was performed with a mixture (30 ml) of 0.5% ropivacaine and 8 mg dexmedetomidine. The efficacy of motor and sensory block was evaluated and the onset time and duration of motor and sensory block were recorded. Venous blood samples were obtained from peripheral vein on the operated side before anesthesia induction (T0), and at 1, 5 and 30 min after tourniquet release (T1-3) to detect the plasma concentrations of MDA and ischemia-modified albumi (IMA). Arterial blood samples were also obtained at the same time points for blood gas analysis. The complications such as nausea and vomiting, respiratory depression, bradycardia and dizziness were recorded. Sufentanil 0.2 μg/kg was given as rescue medication. If the operation could not be completed, general anesthesia was used. Results There was no requirement for rescue analgesics and general anesthesia, and no complications occurred in all the patients. The duration of sensory and motor block was significantly longer, the plasma concentrations of MDA and IMA were significantly lower, and PaO2 and BE were significantly higher in group D than in group C ( P ＜ 0.05). The plasma concentrations of MDA and IMA were significantly higher at T2 and T3 in both groups, the pH value was significantly lower at T1 in group C, PaO2 at T1 and BE at T1 and T2 were significantly lower in both groups than those at T0 ( P ＜ 0.05). Conclusion Dexmedetomidine can not only enhance the efficacy of brachial plexus block with ropivacaine, but also reduce the upper extremity I/R injury caused by tourniquet in patients undergoing upper extremity surgery.