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.

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 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 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.

BACKGROUND: Huperzine A is a reversible cholinesterase inhibitor extracted initially by China from huperzia serrata,a Chinese herb. Its mechanism in improving choline function and ameliorating cognitive state of patients with dementia is to increase acetylcholine (Ach) concentration in neural synapse through impeding the hydrolysis of acetylcholinesterase.OBJECTIVE: To investigate the effects of huperzine A on the improvement of cognitive function of patients with Alzheimer disease (AD).DESIGN: A randomized controlled study based on patients.SETTING: Second internal department of neurology in a general hospital of military area command of Chinese PLA.PARTICIPANTS: Totally 69 AD patients hospitalized in the Second Department of Neurology,General Hospital of Jinan Military Area Command of Chinese PLA between March 2000 and December 2003 were allocated into huperzine A group( n = 38,male,aged between 68 and84 years old,average age of 76 years old) and control group( n = 31,male,aged between 68 and 80years old,average age of 74 years old) according to the wills of the patients.MAIN OUTCOME MEASURES: Positive expression percentage of Fas,Apo2.7 and Bcl-2 on platelet membrane and the integral of mini-mental state examination(MMSE),Hasegawa' s dementia scale(HDS) and activities of daily living (ADL) of two groups after therapy.RESULTS: The positive expression of Fas,Apo2.7 or Bcl-2 on peripheral platelet membrane after therapy in huperzine A group were (2.23 ± 0.49)%,(2.37 ±0. 36)%,or(2.01 ±0. 32)% respectively,which was significantly lower than(3.12 ± 0. 74)%,(2. 83 ± 0.67)% or (2. 59 ± 0. 54)% of control group ( P ＜ 0.01). The prognostic integral of MMSE,HDS or ADL of huperzine A group was 20.45 ±4. 14,21.39 ±4.89,or 36. 15 ±5.11,which was significantly higher or lower than that of the control group(15.76 ± 3.23,17.32 ±2.09,4.26 ±7.21)(P ＜ 0.05).CONCLUSION: HuperzineA actually can effectively improve the memory and cognitive function of AD,and effectively improve ADL as well.

Objective To investigate the effect of MAPK activation on autophagy in the hippocampus of rats with subarachnoid hemorrhage.Methods A total of 100 male SD rats were divided into 5 groups randomly:sham operated group,SAH group,inhibitor U0126 group,inhibitor SB203580 group,SP600125 group.The animal model was established by injecting the autologous blood into cisterna magna twice.The morphological changes of hippocampus nerve cells of rat brain were detected with HE.The mRNA levels of ERK1/2,p38MAPK,JNK and LC3 in hippocampus were detected with quantitative real time PCR and the expression of phosphorylated ERK1/2,phosphorylated p38MAPK,phosphorylated JNK and LC3-Ⅱ in hippocampus of rat brain were detected by immunohistochemistry.Results Compared with the Sham group,the survival rate of neurons in SAH group decreased (6 h:(84.982 ± 5.723) ％,24 h:(74.383± 9.860) ％,48 h:(62.860± 10.820) ％,72 h:(52.260± 10.960) ％) (all P＜0.05).The levels of ERK1/2 mRNA,p38MAPK mRNA,JNK mRNA and LC3 mRNA in hippocampus increased (all P＜ 0.05) and the expression of p-ERK1/2,p-p38MAPK,p-JNK,LC3-Ⅱ proteins increased(all P＜0.05).Compared with the SAH group,the survival rate of neurons in U0126 group was decreased (6 h:(71.620±6.542) ％,24 h:(66.221±7.742)％,48 h:(55.208±8.802) ％,72 h:(46.242±7.782) ％),and the ERK1/2 and LC3 in hippocampus decreased both in mRNA level and in protein level(all P＜0.05).Compared with the SAH group,the survival rate of neurons in SB203580 groups was increased (6 h:(89.082±6.602)％,24 h:(85.840±9.726) ％,48 h:(74.96± 10.916) ％,72 h:(69.211 ± 10.745) ％),and the p38MAPK,LC3-Ⅱ in hippocampus decreased at both mRNA and protein levels (all P＜0.05).Compared with the SAH group,the survival rate of neurons in SP600125 groups was increased (6 h:(91.620± 7.542) ％,24 h:(86.221 ± 10.742) ％,48 h:(75.208±11.802) ％,72 h:(70.242± 11.782) ％).The expression of JNK was decreased while the LC3-Ⅱ was increased in hippocampus (P＜0.05).Conclusion MAPK activation is involved in the autophagy of hippocampal neurons after SAH,in which ERK1/2 activation plays a positive role in the regulation of autophagy in hippocampal neurons after SAH,while p38MAPK and JNK activation plays a negative role in autophagy.

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.

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.