Objective: To investigate the relationship of National Institute of Health Stroke Scale(NIHSS)with heart rate variability(HRV)and cardiac complication in elderly patients with acute cerebral infarction, and to clarify the effect of early drug intervention on the regulation of autonomic nerve function in patients with high NIHSS score. Methods: One hundred twenty-six inpatients with first-onset acute cerebral infarction(ACI)(the cerebral infarction group)and 40 healthy subjects with no history of stroke(the control group)were retrospectively enrolled.All subjects underwent examinations of NIHSS and 24 h dynamic electrocardiogram.According to NIHSS score, patients in the cerebral infarction group were divided into 3 subgroups: NIHSS score 0-4 group(n=32), NIHSS score 5-15 group(n=66)and NIHSS score ≥ 15 group(n=28). The difference in HRV parameters were compared between ACI patients and the controls.Ninety-four ACI patients with NIHSS score ≥5 were randomly divided into 2 groups: (1)the traditional treatment group(n=44), taking routine drugs for cerebral infarction; (2)the special treatment group(n=42), taking metoprolol sustained release tablet and Shensonyangxin capsule as add-on to the routine drugs for cerebral infarction.The 24 h dynamic electrocardiogram examination were conducted 30 days after treatment.The differences in HRV parameters and cardiac complications were compared between the two treatment groups. Results: In patients with acute cerebral infarction, the time-domain parameters of normal-to-normal intervals(NNI), standard deviation of normal-to-normal intervals(SDNN), square root of the mean squared successive differences between normal-to-normaI RR intervals(RMSSD), percentage of adjacent normal-to-normal intervals that differed more than 50 ms(PNN50), frequency domain parameters low-frequency(LF)power and high-frequency(HF)power were significantly reduced as compared with those in the control group(P<0.05). While, pulse rate and frequency domain parameters of sympathetic vagus balance index(LF/HF)were significantly increased(P<0.05). The time-domain parameters(NNI, SDNN, RMSSD and PNN50)and frequency domain parameters(LF, HF)were significantly reduced in the NIHSS score 5-15 group and NIHSS score ≥15 group as compared with the control group and the NIHSS score 0-4 group(P<0.05). The time-domain parameters(RMSSD and PNN50)and frequency domain parameters(LF and HF)were reduced(P<0.05), and the frequency domain parameters(LF/HF)were increased(P<0.05)in NIHSS score ≥15 group compared with in NIHSS score 5-15 group(P<0.05). Compared with pre-treatment, the special treatment group after treatment showed that the time domain parameters(NNI, SDNN, RMSSD and PNN50)and frequency domain parameters(LF and HF)were significantly increased(P<0.05), the frequency domain parameters(LF/HF)were significantly reduced(P<0.05)and the incidence of cardiac complication(total incidence of atrial arrhythmias and ventricular arrhythmias and cardiac complication)were significantly reduced(P<0.05). As compared with the traditional treatment group after treatment, the special treatment group after treatment showed that the time domain parameters(NNI, SDNN, RMSSD and PNN50)and frequency domain parameters(LF and HF)were significantly increased(P<0.05), frequency domain parameters(LF/HF)were significantly reduced(P<0.05)and the total incidence of cardiac complication were significantly reduced(18.4% or 7 cases vs.40.0% or 16 cases, P<0.05). Conclusions The HRV is decreased in patients with acute cerebral infarction.The reduction degree of HRV is more significant along with the higher NIHSS scores, which suggests that patients with high NIHSS scores are prone to an impairment of autonomic nerve function.For patients with high NIHSS scores after stroke, early drug intervention for regulating autonomic nerve function can effectively prevent the occurrence of cardiac emergencies and improve the prognosis of stroke.

[ABSTRACT]AIM:Tostudytheprotectiveeffectofbrain-derivedneurotrophicfactor(BDNF)onvascularendo-thelial cells with H 2 O2-induced oxidative injury .METHODS: Human umbilical vein endothelial cells ( HUVECs ) were cultured in vitro, and the oxidation injury model of HUVECs was established by treatment with H 2 O2 .The oxidatively in-jured HUVECs were cultured with different concentrations (1, 10 and 100μg/L) of BDNF.At the same time, the control group (no injury), PBS treatment after H2O2 injury group and TrkB inhibitor group (with 100 μg/L BDNF and 1∶1 000 TrkB inhibitor) were also set up.The viability of the HUVECs was detected by MTT assay .The levels of LDH, MDA, SOD and GSH were measured .The releases of NO , ET-1 and ICAM-1 were analyzed by ELISA .The changes of ROS pro-duction and cell apoptosis were evaluated by flow cytometry .The protein levels of TrkB , p-TrkB, cleaved caspase-3, Bcl-2 and Bax were determined by Western blot .RESULTS:Compared with uninjured control group , in H2 O2 oxidative injury plus PBS treatment group , the viability of the cells was decreased significantly , the LDH and MDA levels were increased significantly and the activities of SOD and GSH were decreased significantly .The NO secretion was decreased , and the ET-1 and ICAM-1 concentrations were increased significantly .The ROS content and apoptotic rate were increased significantly . The protein levels of cleaved caspase-3 and Bax were increased but Bcl-2 protein expression was decreased significantly . Compared with PBS treatment group , in H2 O2-injured HUVECs treated with different concentrations of BDNF , the cell via-bility was gradually increased , the LDH and MDA levels were decreased and the activities of SOD and GSH were increased gradually .The secretion of NO was increased but ET-1 and ICAM-1 were decreased gradually .The ROS content and apop-totic rate were decreased significantly .The TrkB and p-TrkB levels were significantly increased significantly , the protein expression of cleaved-caspase 3 and Bax was decreased gradually and the Bcl-2 protein expression increased gradually .The role of BDNF was inhibited by TrkB inhibitor .CONCLUSION:BDNF protects HUVECs from oxidative injury by binding with TrkB to activate the BDNF-TrkB signaling pathways .

Objective To investigate the effects of brain-derived neurotrophic factor (BDNF) pretreatment on H9c2 myocardial hypoxia/reoxygenation (H/R) injury, and explore its mechanism. Methods The H9c2 myocardial cells were cul?tured in vitro and (95%O2+5%CO2) oxygen cultured 12 h after (95%N2+5%CO2) hypoxia cultured 4 h to establish the H/R model. The cells were divided into normal control group, H/R group, different concentrations (1, 10, 100μg/L) BDNF pre?treatment in H/R groups and TrkB-inhibitor group (with 100μg/L BDNF and 1∶1 000 TrkB inhibitor pre-treatment in H/R group). The cell survival rate was measured by MTT method in different groups. The lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA) and superoxide dismutase (SOD) content and activity were detected after H/R injury. The apoptotic rate of H9c2 myocardial cells were detected by flow cytometry, and the expressions of TrkB, Bcl-2 and Bax protein were detected by Western blot assay. Results Compared with the normal control group, the survival rate of H9c2 myocardial cells was decreased significantly in H/R model group (P < 0.05), LDH, CK and MDA contents were increased and SOD activity was decreased (P<0.05). The cell apoptosis rate was increased significantly (P<0.05). The anti-apoptosis Bcl-2 protein expression was decreased, pro-apoptosis Bax protein expression was increased in H/R model group (P<0.05). Compared with the H/R model group, the cell survival rates of H9c2 myocardial cells were increased after pre-treatment with different concentrations of BDNF (P<0.05);LDH, CK and MDA contents were decreased and SOD activity were in?creased respectively (P < 0.05). The cell apoptotic rates were decreased (P < 0.05). The expressions of TrkB receptor and Bcl-2 protein gradually increased, while the expression of Bax protein was gradually decreased (P<0.05). The role of BDNF was inhibited by TrkB inhibitor. Conclusion BDNF pre-treatment can promote the cell survival rate of H9c2 myocardial cells after H/R injury, which plays a protective role by inhibiting the cell apoptotic rate and maintaining antioxidant capacity, and associates with BDNF-TrkB signaling pathways.

Objective: To investigate the effect of tanshinone IIA (TSN) on left ventricular hypertrophy (LVH) and cardiomyocyte apoptosis in spontaneous hypertensive rats (SHRs). Methods: A total of 60 SHRs at 8 weeks of age were randomly divided into 3 group: Blank control group, the rats were sacriifced at 8 weeks, TSN group, the rats were treated with TSN at 1 ml/(kg?d) for 18 weeks and Solvent control group, the rats were treated with the solvent at 1 ml/(kg?d) for 18 weeks. n=20 in each group and 15 rats were used for the experiments. The systolic blood pressure (SBP) and left ventricular mass index (LVMI) were examined, cardiomyocyte’s diameter and surface area were measured by HE staining, the apoptosis rate was evaluated by TUNEL method and the apoptosis related protein expression s of Bcl-2, Bax and p53 were determined by Western blot analysis. Results: ①Compared with Solvent control group, TSN group had decreased LVMI (3.23 ± 0.24) mg/g vs (4.58 ± 0.68) mg/g,cardiomyocyte’s diameter (16.13 ± 1.77) μm vs (27.15 ± 3.52) μm and surface area (230.23 ± 69.37) μm2 vs (490.12 ± 118.96) μm2and decreased apoptosis rate (7.45 ± 1.78) % vs (10.61 ± 2.77) %, allP<0.01.②With NAPDH reference correction, compared with Solvent control group, TSN group presented increased protein expression of Bcl-2 (0.97 ± 0.31) vs (0.40 ± 0.11) and decreased Bax (0.37 ± 0.15) vs (1.81 ± 0.44), decreased p53 (0.83 ± 0.18) vs (2.72 ± 0.28), allP<0.05 or P<0.01. The above indexes were similar between TSN group and Blank control group,P>0.05. Conclusion: TSN could inhibit the development of LVH and decrease the cardiomyocyte apoptosis, which might be via up-regulating the protein expressions of Bcl-2 and down-regulating Bax and p53 in SHRs.

Objective To prepared inclusion complex of β-cyclodextrin (β-CD) and its derivatives (methyl -β-cyclode-xtrin, 2,6-dimethyl -β-cyclodextrin, hydroxypropyl -β-cyclodextrin ) with entrcavir, and explore effect of different inclusion complex on the solubility and dissolution of entecavir.Methods Regression equation was established,prepared the inclusion complexes by a magnetic stiring method,grinding method and ultrasonic method.While inclusion rate was an index,the effects of reaction process was inquired by mole ratio, inclusion temperature, inclusion time and stirring rate and the most suitable method and technology of inclusion was optimized.Determinated the solubility with HPLC method and investigated in vitro dissolution with a small cup method.Results The regression equation was A=1.026 ×106C+4248.8, R2 =0.9999 (n=6), in the range of 0.497~4.97 g/mL, linear relationship was good.The best inclusion method was magnetic stirring, the best process conditions were: mole ratio was 1 ︰1, stirring speed was 300 r/min, inclusion time was 4 hours, inclusion temperature was 50 ℃.The four inclusion complexes all were white loose powder materials.The solubility of ETV,β-CD-ETV,HP-β-CD-ETV,RM-β-CD-ETV,2,6-DM-β-CD-ETV were 2.51, 13.4, 18.9, 37.6 and 89.5 g/L.The dissolution rates was ETV<β-CD-ETV

Objective To observe the effect of integrated therapy of Chinese medicine and chemical drugs on adverse reaction and curative effect of initial treatment of secondary pulmonary tuberculosis. Methods Totally 1404 patients with secondary pulmonary tuberculosis and TCM lung consumption diagnostic criteria (syndrome of lung yin deficiency, qi-yin deficiency, yin-deficiency caused excessive fire) were chosen for single blind, randomized, controlled, multicenter clinical trials. Trial group was given 2HRZE/4HR, 1 time/day with Chinese medicine 2 or 3 times/day, and control group was given 2HRZE/4HR only for six months. The adverse reactions and clinical symptoms were observed to evaluate clinical efficacy and safety. Results In terms of reducing liver damage and other adverse reactions, the ratio of trial group had statistical difference with that of control group (P<0.001). In symptom scores of lung yin deficiency syndrome treated for 2, 4, 6 months, yin-deficiency caused excessive fire syndrome treated for 6 months, qi-yin deficiency syndrome treated for 4, 6 months, the differences between the two groups were significant (P<0.001). TCM syndrome curative effect between the two groups was statistical different (P<0.001). Safety evaluation result between the two groups was statistical different by tratified analysis (P<0.001). Conclusion Integrated therapy of Chinese medicine and chemical drugs can improve the symptoms and reduce adverse reactions caused by chemical drugs. It can enhance the curative effect and safety.

To explore the role and possible mechanism of apoptosis and caspase-3 activity in the development of multicellular drug resistance of ovary cancer. Ovarian cancer cell A2780 multicellular spheroids (MCS) were obtained from three-dimensional culture. Drug sensitivity of monolayer cells (MC) and MCS were respectively tested by MTT staining and cytometry. The apoptosis of MC and MCS were determined by the flow cytometry (FCM). The expression of bcl-2 and caspase-3 in A2780/MC and A2780/MCS were detected by using Western blot and caspase-3 assay kit. A2780/MC was compacted into mass after 2 days in three-dimensional cell culture model, and MCS had more than two layers of cells growing within 5 days. Compared with A2780/MC, A2780/MCS were more resistant to the anticancer drug, and the apoptosis rate was significantly lower than those of A2780/MC. The activity of caspase-3 in A2780/MCS was significantly lower than the A2780/MC. But the expression of bcl-2 in A2780/MCS was significantly higher than that in A2780/MC. It was suggested that the drug resistance of MCS might be associated with the overexpression of anti-apoptosis protein bcl-2 and the down-regulation of caspase-3 activity.

To explore the role and possible mechanism of apoptosis and caspase-3 activity in the development of multicellular drug resistance of ovary cancer. Ovarian cancer cell A2780 multicellular spheroids (MCS) were obtained from three-dimensional culture. Drug sensitivity of monolayer cells (MC) and MCS were respectively tested by MTT staining and cytometry. The apoptosis of MC and MCS were determined by the flow cytometry (FCM). The expression of bcl-2 and caspase-3 in A2780/MC and A2780/MCS were detected by using Western blot and caspase-3 assay kit. A2780/MC was compacted into mass after 2 days in three-dimensional cell culture model, and MCS had more than two layers of cells growing within 5 days. Compared with A2780/MC, A2780/MCS were more resistant to the anticancer drug, and the apoptosis rate was significantly lower than those of A2780/MC. The activity of caspase-3 in A2780/MCS was significantly lower than the A2780/MC.But the expression of bcl-2 in A2780/MCS was significantly higher than that in A2780/MC. It was suggested that the drug resistance of MCS might be associated with the overexpression of anti-apoptosis protein bcl-2 and the down-regulation of caspase-3 activity.