Myocardial ischemia/reperfusion(MI/R) injury refers to restoring blood perfusion after myocardial ischemia for a relatively long time, but the tissue appears more obvious and severe myocardial damage and dysfunction than before reperfusion, this phenomenon mainly relates to inflammation, oxidative stress, cell apoptosis, cell autophagy and so on.NF-κB is a nuclear transcription factor involved in regulating a variety of pathophysiological processes, which also plays an important role in all aspects of MI/R injury.The in-depth study of NF-κB in MI/R injury and clinical application of related research will provide new ideas and methods for treating MI/R injury.

Objective To investigate the effect of valsartan and amlodipine on urinary microalbumin in elderly hypertensive patients. Methods 100 elderly patients with hypertension treated in our hospital from May 2015 to October 2016 were selected and randomly divided into the control group and the experimental group, with 50 patients in each group. The patients in the control group received oral valsartan, and the patients in the experimental group were treated with valsartan and amlodipine. The treatment time of the experimental group and the control group was 12 weeks. The clinical indexes of the experimental group and the control group were compared and analyzed. Results After the corresponding treatment, the experimental group and the control group did not have obvious adverse reactions. There were 2 cases of headache in the experimental group, 1 cases of vertigo, and 2 cases of vertigo in the control group. However, there was no significant difference in the incidence of adverse reactions between the experimental group and the control group, and there was no statistical significance. The antihypertensive effect of the experimental group was significantly higher than that of the control group, with statistical significance (P<0.05). After treatment, the urinary microalbumin in the experimental group and the control group was significantly lower than that in the treatment group, and the level of microalbuminuria in the experimental group was lower than that in the control group, with statistical significance (P<0.05). Conclusion The clinical effect of treatment of elderly patients with hypertension better combined with valsartan and amlodipine, antihypertensive effect is stronger, can significantly improve the patient's urinary albumin, with further clinical promotion and application significance.

Objective To study the effects of 3-n-butylphthalide (NBP) on motor function and learning and memory ability in rats with diffuse brain injury (DBI). Methods 128 male Sprague-Dawley rats were randomly divided into sham operation group, traumatic group,low-dose NBP treatment group and high-dose NBP treatment group with 32 rats in each group. Every group was divided into 24 h, 48 h and 72 h subgroups. DBI rat model was established according to the description of Marmarou's diffused brain injury. The histopathologic changes of cerebral tissue (0.2 cm bilateralis coronal line) were observed by light and electron microscope at 24 h, 48 h and 72 h after injury. Morris water maze and rolling test were performed daily at 24, 48 and 72 h (time points). Results Compared with the model group, the damage of brain tissue decreased, and the survival nerve cells increased (P<0.01). The behavioral tests showed that the latency to find the platform shortened (P<0.01) and the frequency of crossing the platform increased in the 72 h subgroup (P<0.01). The general movement ability enhanced in NBP groups (P<0.05). All the indexes were more significant in high-dose NBP treatment group. Conclusion NBP can improve neurological function and learning and memory function after brain injury and the molecular mechanisms is related to the decrease of the nerve cells lose and traumatic cerebral ultrastructure damage.

Objective:To study the effect of endothelin receptor antagonist BQ-123 on the nerve function damage after subarachnoid hemorrhage (SAH)in the rats,and to explore the mechanisms.Methods:Total 120 male SD rats were divided into sham group,SAH group,low dose of BQ-123 group (50 μg· kg-1 )and high dose of BQ-123 group (75 μg·kg-1 ).The SAH rat models were established by injecting the autologous blood into cisterna magna twice.The morphological changes of hippocampus nerve cells of rat brain tissue were detected with HE staining, and the expressions of mTOR, Beclin-1 and LC3-Ⅱ in the hippocampus of rats were detected with immunohistochemistry and RT-PCR;the shuttle-box experiment was used to evaluate the abilities of learning and memory,and the holding power evaluation was used to evaluate the forelimb pulling force of the rats in various groups at each time point.Results:Compared with sham group,the morphological damages of neurons of the rats in SAH group were increased,the survival rate of neurons of the rats in SAH group was decreased (P <0.05),the expression levels of mTOR mRNA,Beclin-1 mRNA and LC3-Ⅱ mRNA in hippocampus tissue of the rats were increased (P < 0.05),and the abtilities of learning and memory and the values of holding power were decreased (P <0.05).Compared with SAH group,the morphological damages of neurons of the rats in BQ-123 groups were decreased,the survival rates of neurons of the rats in BQ-123 groups were increased (P < 0.05),the expression levels of mTOR mRNA of rats were decreased (P <0.05),the expression levels of Beclin-1 mRNA and LC3-ⅡmRNA in hippocampus tissue were increased (P <0.05),and the abilities of learning and memory and the values of holding power were increased (P < 0.05 ). The changes were more significant in high dose of BQ-123 group compared with low dose of BQ-123 group (P <0.05).Conclusion:BQ-123 can improve the nerve function damage after SAH in the rats,its mechanism may be related to regulating the mTOR/autophagy signaling pathway.

Objectives To investigate the treatment effect of endothelin A receptor antagonist BQ-123 on early brain injury of subarachnoid hemorrhage (SAH ) in rats and its mechanism. Methods According to the random number table method,120 SD rats were divided into four groups:a sham operation (sham),a SAH,a high-dose BQ-123 (75 μg/ kg),and a low-dose BQ-123 (50 μg/ kg)(n = 30 in each group). A rat model was induced by using the injection of blood into cisterna magna twice. After establishing models at hours 6,24,72,and 144,the rats were further divided into four subgroups. Light and electron microscopes were used to observe the changes of the morphological structure in hippocampal area. Immunohistochemistry and RT-PCR were used to detect the expression levels of phosphoinositide 3 kina (PI3-K),protein kinase B (PKB/ Akt),and mammal target of rapamycin (mTOR). Results (1)In the process of model making,7 rats died and 1 model did not meet the criteria and was excluded from the SAH group. Six rats died in the high-dose BQ-123 group and the low-dose BQ-123 group respectively. One rat in each group did not meet the criteria and was excluded. The rats were included in the final statistical analysis:30 in the sham group,22 in the SAH group,23 in the high-dose BQ-123 group,and 23 in the low-dose BQ-123 group. (2)Compared with the sham group,the expression levels of PI3-K,AKt and mTOR were increased signifi-cantly (all P < 0. 05). Compared with SAH group,the hippocampal neuronal morphology and structure damage were alleviated in the low-dose BQ-123 group. The number of surviving neurons at each time point was increased ([132 ±18],[110 ±16],[84 ±13],[92 ± 10]cells/ HP,all P < 0. 05). The tensile force values of rats were increased at each time point and the learning and memory function were improved. The expression levels of PI3-K and Akt were further increased (all P < 0. 05). The expression level of mTOR was decreased (all P < 0. 05). (3)Compared with the low-dose BQ-123 group,the morphological and structural damage of hippocampal neurons were alleviated. The number of surviving neurons at each time point ([153 ±20],[131 ± 18],[137 ±19]and [135 ± 17]cells/ HP)was increased (all P < 0. 05). The tensile force values of the rats were increased at each time point. The learning and memory function of the animals were improved. The expression levels of PI3-K (3. 8 ± 0. 8,8. 9 ± 2. 4,8. 6 ± 2. 4,and 6. 2 ± 2. 0)and Akt (4. 86 ± 1. 74, 8. 64 ± 1. 62,7. 94 ± 1. 70,and 6. 48 ± 1. 58)were further increased (all P < 0. 05). The expression levels of mTOR (2. 89 ± 0. 26,2. 14 ± 0. 18,1. 94 ± 0. 17,and 1. 62 ± 0. 12)were decreased (all P < 0. 05). Conclusions BQ-123 has as a good therapeutic effect for early brain injury after SAH. Its mechanism may be associated with the regulation of PI3-K/ Akt signaling pathway.

Objective To observe the effect of exposure of emphysema with intermittent hypoxia on oxidative stress injury of myocardial cells in rats. Methods Sixty male Wistar rats were divided randomly into four experimental groups(each n=15). The normal control group was bred normally. The emphysema group was exposed to cigarette smoke twice a day(once 30 minutes). The intermittent hypoxia(IH)group was exposed to intermittent hypoxia circumstance 8 hours/day,and the emphysema with IH group was exposed to cigarette smoke twice a day (once 30 minutes)and intermittent hypoxia circumstance 8 hours/day. Each group was exposed for 8 weeks. At the beginning of 9 weeks,the blood gas analysis was performed in 5 rats selected randomly from each group,and the rest rats were sacrificed and their hearts and lungs were taken. Under light microscope,the lung tissues stained with hematoxylin-eosin(HE)were examined. The lung pathology and the results of blood gas analysis showed that the emphysema with IH rat model was established successfully. The levels of malonaldehyde(MDA)and superoxide dismutase(SOD)in rat myocardium were measured by enzyme-linked immunosorbent assay(ELISA),and the subunit p22phox mRNA expressions of nicotinamide adenine dinucleotide phosphate(NADPH)-oxidase were detected by real-time reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with the normal group, the MDA levels and p22phox mRNA expressions were increased obviously in emphysema group, IH group and emphysema with IH group〔MDA(μmol/g):2.93±0.54, 3.58±0.63, 4.51±0.72 vs. 1.75±0.56, p22phox mRNA:0.043±0.004,0.067±0.015,0.123±0.016 vs. 0.018±0.002,all P<0.05〕,but the activities of SOD were decreased significantly(U/mg:36.07±4.79,33.51±7.12,24.29±5.36 vs. 46.08±5.12,all P<0.05). In emphysema with IH group,the increase of MDA levels and p22phox mRNA expressions and decrease of SOD levels were more remarkable compared with those in emphysema group and IH group(all P<0.05). The expression of p22phox mRNA was positively correlated with MDA level(r=0.734,P<0.001). Conclusion The myocardial tissue oxidative stress injury in rats induced by emphysema with intermittent hypoxia exposure is more serious than that induced by exposure of either emphysema or intermittent hypoxia alone,NADPH oxidase possibly being the important medium of myocardial cell response to oxidative stress.

AIM: To construct signal peptide-canstatin expression vector pEGFP-C1-SP-Can and express secretable mouse canstatin fusion protein in Eca-109 cells.METHODS: Site-directed mutagenesis was used in amplifying the signal peptide of murine plasminogen to construct the plasmid pEGFP-C1-SP.The cDNA of mouse canstatin,obtained from a cloning vector pMD18T-Can by PCR,was inserted into pEGFP-C1-SP to construct a secretable expression vector pEGFP-C1-SP-Can.Constructed plasmid pEGFP-C1-SP-Can was transiently transfected into Eca-109 cells via lipofectamine,and subsequently its secretable expression in the medium of cultured Eca-109 was observed by Western blotting.RESULTS: DNA sequencing and restriction enzyme analysis attested the validity of the constructed plasmids pEGFP-C1-SP and pEGFP-C1-SP-Can.EGFPcanstatin fusion protein was proved to be secretably expressed in Eca-109 by Western blotting.CONCLUSION: It is concluded that the constructed vector pEGFP-C1-SP-Can is valid and capable of expression in Eca-109,these findings provide a basis for testing the function of mouse canstatin and its application in gene therapy.

AIM: To clone and express mouse canstatin (m canstatin)cDNA and provide a basis for the further research on its anti-angiogenic activity and potential application for cancer therapy. METHODS: Total RNA was extracted from mouse liver tissue by Trizol Reagent, and mouse canstatin cDNA was amplified by RT- PCR, then cloned into vector pMD18-T for sequencing. pET30a(+)-m canstatin recombinant plasmid was constructed and expressed in E.coli BL21 with induction of IPTG. RESULTS: Mouse canstatin cDNA is 684 bp coding 227 amino acids. The sequences of both cDNA and amino acid share high homology with human canstatin, with cDNA identity at 89% and amino acids identity at 96% to human canstatin. In the present study, pET30a(+)-m canstatin recombinant plasmid was expressed in E.coli BL21. CONCLUSION: Mouse canstatin cDNA has been cloned for the first time. Constructed pET30a(+)-m canstatin recombinant plasmid is highly expressed in E.coli BL21.

To investigate the effects of intrinsic nitric oxide (NO) on the expression of interleukin-4 (IL-4) mRNA and interferon-γ (IFN-γ) mRNA in the airway inflammation of asthma,the rat models of asthmatic inflammaiton were established by sensitizing and then challenging the animals with ovalbumin. The 24 animals were randomly divided into control group, sensitized group, sensitized and L-Arg-treated group as well as L-NAME-treated group equally. By using in situ hybridization combined with compute physiological quantitative imaging analysis techniques,the influence of intrinsic NO on the expression of IL-4 mRNA and IFN-γ mRNA in the airway inflammatory cells was observed. In situ hybridization study demonstrated that IL-4 mRNA expression was obviously increased as compared with that in the control group, mainly distributed in the inflammatory cells in the submucous of airways in the sensitized group. The increase of intensity of IL-4 mRNA expression was positively correlated with the numbers of eosinophil (Eos) and lymphocyte (both with P＜0.05) in the sensitized group. There was no statistically difference in IFNγ expression between the control group and the sensitized group. Imaging analysis showed that LNAME could inhibit the expression of IL-4 mRNA (P＜0.05) and increase the expression of IFNγ mRNA (P＜0.05), while L-Arg could increase the expression of IL-4 mRNA in inflammatory cells (P＜0.05). It was indicated that a suitable levels of intrinsic NO can influence the expression of IL-4 mRNA of Th2 lymphocytes and the expression of IFN-γ mRNA of Th1 lymphocytes and in turn, promote the development of asthmatic airway inflammation.

To investigate the effects of intrinsic nitric oxide (NO) on the expression of interleukin-4 (IL-4) mRNA and interferon-γ (IFN-γ) mRNA in the airway inflammation of asthma,the rat models of asthmatic inflammaiton were established by sensitizing and then challenging the animals with ovalbumin. The 24 animals were randomly divided into control group, sensitized group, sensitized and L-Arg-treated group as well as L-NAME-treated group equally. By using in situ hybridization combined with compute physiological quantitative imaging analysis techniques,the influence of intrinsic NO on the expression of IL-4 mRNA and IFN-γ mRNA in the airway inflammatory cells was observed. In situ hybridization study demonstrated that IL-4 mRNA expression was obviously increased as compared with that in the control group, mainly distributed in the inflammatory cells in the submucous of airways in the sensitized group. The increase of intensity of IL-4 mRNA expression was positively correlated with the numbers of eosinophil (Eos) and lymphocyte (both with P＜0.05) in the sensitized group. There was no statistically difference in IFNγ expression between the control group and the sensitized group. Imaging analysis showed that LNAME could inhibit the expression of IL-4 mRNA (P＜0.05) and increase the expression of IFNγ mRNA (P＜0.05), while L-Arg could increase the expression of IL-4 mRNA in inflammatory cells (P＜0.05). It was indicated that a suitable levels of intrinsic NO can influence the expression of IL-4 mRNA of Th2 lymphocytes and the expression of IFN-γ mRNA of Th1 lymphocytes and in turn, promote the development of asthmatic airway inflammation.