Objective To study the role of caspase 3 in ischemic brain damage of rats, and further understand the molecular mechanisms of ischemic cerebral vascular disease.Methods Rat models of the left middle cerebral artery (MCA) occlusion/reperfusion were made using a modification of the intraluminal sature method of Longa established by Belayev, infarct zones were confirmed by 2,3,5 triphenyltetrazolium chloride (TTC) staining, and caspase 3 expression on brain sections at the mRNA and active protein level was detected with in situ hybridization and immunohistochemistry technique, respectively.Results After 2 hours of left MCA ischemia followed by 24 hours of reperfusion, obvious infarct in the MCA dominate regions was confirmed by TTC staining; low levels of caspase 3 mRNA, and fewer of its active protein expression was found in normal brains, sham brains and contralateral brains of MCAO rats; both caspase 3 mRNA and activated protein expression in ipsilateral region were increased after 24 hours of recirculation, and even higher levels were detected at 48 hours of reperfusion.Conclusion Apoptotic mechanism might involve in delayed neuronal death after cerebral ischemia, and caspase 3 might play an important role in ischemic neuronal injury.

Chlorides ; metabolism ; Diarrhea ; congenital ; Humans

Objective:To investigate whether the habenula takes part in the depressor effect of losartan.Methods:Arterial blood pressure,heart rate and the unit dischargings of habenular neurons in rat were recorded simultaneously.Results:Arterial blood pressure was apparently decreased by 10 mg/kg losartan intraperitoneally(ip),but heart rate did not change;63.64% (21/33) unit dischargings of habenular neurons was increased in the rate of discharges.Conclusion:The depressor effect of losartan (ip) may be involved in the excitation of habenular neurons by lsoartan(ip).

Objective To investigate the characteristics of marital quality and social support status of the patients with breast cancer during the period of chemotherapy and the correlation of both factors.Methods Seventy-seven patients with breast cancer were divided into beginning-, middle-and final-staging groups of chemotherapy.And they were studied with Olson marital quality questionnaire and Social Support Rating Scale.Results The total scores of marital quality and other factors were the lowest in the mid-chemotherapy group.Among three groups,the mean of the objective support and availability were gradually increased with the chemotherapy going.The score of objective support was higher in the final than beginning chemotherapy groups (P =0.009).There was a positive correlation between marital satisfaction and the objective support (r =0.248, P =0.036), and between the total score of quality of marriage and subjective support (r =0.286, P = 0.015).Conclusion The marital quality of breast cancer patients receiving chemotherapy is poor,and subjective or and objective support might improve the patient's marital quality.

Objective To explore the clinical significance of the precision evaluation of computed tomography angiography and color Doppler image formation technology to the condition of the vascular system in the donor site.Methods From January,2014 to December,2014,CTA and color Doppler were used to examine conditions of the first dorsal metatarsal artery's courser and the size of original caliber in 25 patients with thumb or finger defected.Also the distance between skin and the first dorsal metatarsal artery.The accurate positions and adjacent relations of vessels in donor site were precisely marked and then the vessel original caliber was measured.Results All the first dorsal metatarsal arteries of 25 cases can be appeared clearly in CTA and color Doppler.Blood vessels line and filling results were completely consistent in intraoperative findings.The difference of the vessel original average caliber was statistically significant (P ＜ 0.05) (1.20 ± 0.25 mm,1.41 ± 0.15 mm and 1.3 ±0.2 mm in Doppler,CTA and operation,respectively) while the distance was not significant (P ＞ 0.05)(9.20 ±3.06 mm,10.32 ± 2.76 mm and 0.2 ± 2.6 mm in Doppler,CTA and operation,respectively) All the 25 transplanted toes were survived.Conclusion CTA combined color Doppler image formation technology in thumb or finger reconstruction can objectively reflect the real situation of the vessels in donor sites,and the structural characteristics and classification of anatomy for individualized,and has important clinical value in preoperative foot blood supply assessment.

Objective: To study caspase-3 activities and the neuroprotective effect of caspase-3 inhibitor Ac-DEVD-CMK in primary cultures of rat hippocampal neurons during hypoxia/reoxygenation (H/R). Methods: After pretreated with Ac-DEVD-CMK or the vector DMSO fo 1 h, primary cultures of rat hippocampal neurons were induced to hypoxia for 3 h, followed by 12 to 48 h of reoxygenation. The neuronal viability was estimated by MTT assay, and caspase-3 cellular activities were measured by a colorimetric method using Ac-DEVD-pNA as a substrate. Results: During H/R, the cultured rat hippocampal neuronal viability gradually decreased, and caspase-3 activities in the primary cultures of rat hippocampal neurons were significantly increased, and peaked at 24 h of reoxygenation. Caspase-3 activities were decreased in the neurons pretreated with Ac-DEVD-CMK in a dose dependent manner. Conclusion: Delayed neuronal death is detected in cultured hippocampal neurons during H/R, and apoptosis mediated by caspase-3 may play an important role in it, and caspase-3 inhibitor has a neuroprotective effect on them.

Objective: To establish a method of restriction fragment length polymorphism (RFLP) to determine the origin of sika deer bones.Methods: The DNA in the bone samples was extracted after decalcification, and then amplified using polymerase chain reaction (PCR).The origin of the samples was further identified using RFLP analysis.Results: The bone samples of sika deer and red deer could be distinguished from those of pig, bovine and dog by PCR.And the samples of sika deer and red deer could be further distinguished by RFLP through the analysis of the length of restriction enzyme XbaI.Conclusion: A RFLP method is established to determine the origin of sika deer bones.

Background Toll-like receptor 4 (TLR4) is an immune related receptor.It plays an important role in inducing inflammation response.The inflammatory response secondary to optic nerve crush will results in serious retinal damage.It is worthy of studying the expression and effect of TLR4 in retina after optic nerve crush.Objective This experimental study was to explore the role of TLR4 in the loss of retinal ganglion cells(RGCs) after optic nerve crush.Methods Twenty-four SPF adult health Sprague-Dawley (SD) rats were used in the study and radomized into two groups based to the experimental time.The optical nerve crush models were established by crushing the optical nerve in the right eyes of the rats,and the left normal eyes served as controls.The rats were sacrificed by over anesthesia and retinas were isolated 3 days and 7 days after operation.Expression of TLR4 in the retinas was detected using immunofluorescence method.Reverse trancription PCR (RT-PCR) and Western blot were applied respectively for the detection of TLR4 mRNA and protein in the retinas.The apoptosis of RGCs was evaluated using TUNEL staining.The use and care of experimental animals followed theGuide for the Care and Use of Laboratory Animals of NIH.This study was approved by the Institutional Animal Care and Use Committee at the Zhongshan Ophthalmic Center.Results The expression of TLR4 in rat retinas presented with green fluorescence mainly in the inner layer of retinas.The fluorescence was enhanced in the model 3 days group and the model 7 days group compared with the corresponding control groups.The relative expressing values of TLR4 mRNA in the retinas were 2.92±0.06and 3.92±0.12 in the model 3 days and 7 days groups,respectively,which were significantly higher than 2.87±0.12and 3.44±0.17 in the control 3 days and 7 days group (t3d =-12.888,P＜0.001 ;t7d =-4.669,P=0.010).In the model 3 days group and model 7 days group,the grey values of TLR4 protein were 1.14±0.05 and 1.49±0.03,and those in the control 3 days and 7 days groups were 0.99 ± 0.09 and 1.38 ± 0.07,showing significant differences between them(t3d =-11.324,P＜0.001 ;t7d =-5.638,P=0.005).Apoptotic RGCs were obviously increased in the optic nerve damage group in comparison with the control group.Conclusions The TLR4 is over-expressed in the inner layer of retina after optic nerve crush,which suggestes that TLR4 is probably involved in the loss of RGCs after optic nerve crush.

Objective To investigate the association between β2-adrenergic receptor gene polymorphism and coronary atherosclerotic heart disease, and to provide reference for clinical disease prevention and treatment.MethodsThe clinical data of 200 patients with coronary atherosclerotic heart disease confirmed by coronary angiography were selected and included in the study group, and 200 healthy subjects without coronary atherosclerotic heart disease group.The polymorphism of β2 adrenergic receptor gene was detected and the frequency of each gene was analyzed.ResultsThe genotype frequencies of β2-adrenergic receptor gene were in accordance with Hardy-Weinberg equilibrium, the difference was not statistically significant.According to dominant genetic model, the frequency of AA+AG was 46.0% vs 58.0% lower than that of the control group, and had statistical significance, The genotype frequency of GG genotype in study group was significantly lower than that in control group 14.0% vs 26.0%, χ2=26.20, P=0.00.The frequency of GG genotype in study group was significantly higher than that in control group 54.0% vs 42.0%, χ2=5.76, P=0.01.The frequency of A gene was 38.0% compared with 44.0% in control group, χ2=1.48, P=0.22;the frequency of AA gene in study group was 30.0%, and the frequency of gene A was 38.0%, compared with 56.0% Compared with 32.0% in the control group, χ2=0.18, P=0.66.ConclusionThe A/G polymorphism of β2-adrenergic receptor gene is closely related to the clinical pathogenesis of coronary atherosclerotic heart disease, and the A allele may be a protective factor in patients with coronary atherosclerotic heart disease.

Objective To investigate the effects ofdocosahexenoic acid (DHA) on large conductance Ca2+-activated K+ (BK) channels in normal retinal artery smooth muscle cells (RASMCs).Methods Cultured human RASMCs (6 th-8 th generations) were used to patch clamp experiment.The open probabihties (NP0) in BK channels with different concentrations (0.0,1.0,3.0,5.0,7.5,10.0 μmol/L) of DHA were recorded by patch clamp technique in single channel configuration.RASMCs were intervened by different concentrations (0.0,1.0,5.0 μmol/L) of DHA as control group,low and high doses of DHA groups,respectively.The protein expressions of β subunit of BK channels in RASMCs from three groups were measured by Western blot.Results The NP0 of BK channels were 0.044 4±0.001 2,0.081 2±0.004 2,0.209 0±0.006 1,0.310 5±0.005 3,0.465 0±0.007 8 and 0.497 7±0.014 5 with perfusate of 0.0,1.0,3.0,5.0,7.5,10.0 μtmol/L DHA.DHA activated BK channels in a dose-dependent manner (F=2.621,P＜0.05).There was no significant difference in the protein expression of control group,low and high doses of DHA groups (F=1 1.657,P＞0.05).Conclusion DHA can directly activate BK channels,no increasing in subunit expression of BK channels.