Abstract:[Abstract] Objective To construct and identify miR-1 adenovirus vector, and to analyze its effect on cardiac hypertrophy. Methods The primers of miR-1 precursor were designed for PCR amplification, and the PCR products were cloned into adenovirus shuttle plasmid pAdTrack and linearized by enzyme PmeⅠ; the resultant plasmid was co-transfected into E. coli BJ5183 cells with adenovirus backbone plasmid pAdEasy-1 for homologous recombination. Then the recombinant plasmid was identified, linearized and packaged into QBI-293A cells to amplify the recombinant adenovirus Ad-miR-1, which was then used to infect cardiomyocytes. Real-time quantitative PCR was used to observe the expression of miR-1 and two hypertrophic markers, the atrial natriuretic peptide(Nppa) and β-myosin heavy chain (myh7),in cultured primary cardiomyocytes. Cell surface area was analyzed using software AxioVision 4.7.1 (Carl Zeiss). Methods Sequencing and enzyme digestion showed that the miR-1 recombinant plasmid was successfully constructed. Real-time quantitative PCR confirmed that adenovirus Ad-miR-1 significantly enhanced intracellular miR-1 expression in cardiomyocytes and reduced cell surface area and the expression of Nppa and myh7. Methods The adenovirus expressing miR-1 has been successfully constructed and it can be transfected into cardiomyocytes to increase the expression of miR-1, thus inhibiting cardiomyocyte growth.

Abstract:\[Abstract\] Objective To investigate the possible mechanism by which PRKAG2 G100S, a novel mutation recently found in a Chinese family with WolffParkinsonWhite(WPW) syndrome, induces cardiac hypertrophy. Methods The recombinant adenovirus containing human PRKAG2 G100S was constructed using Invitrogen's Gateway TM system and was used to infect rat embryonic H9c2 cardiomyocytes and cardiomyocytes of neonatal SD rats using EGFP as the reporter gene; Western blotting analysis was used to determine PRKAG2 protein expression. The intracellular free Ca2+ was determined in the H9c2 cells before and 48 h after infected with AdEGFP, AdPRKAG2 or AdPRKAG2 G100S by incubating with Rohd2/AM. The glycogen contents in neonatal SD rat cardiomyocytes were determined by PAS 4872 h after infection. Results Bright green fluorescence was observed in the cultured H9c2 cells and neonatal SD rat cardiomyocytes 48 h after infected with AdPRKAG2 G100S, and PRKAG2 protein expression was identified with PRKAG2 monoclonal antibody. Compared with wildtype PRKAG2 and EGFP groups, intercellular free Ca2+ concentration in H9c2 cells of AdPRKAG2 G100S group had no noticeable changes 48 h after infection, and the concentrations were not significantly different in AdPRKAG2 G100S group before and 48 h after infection. PAS revealed evident glycogen storage in the neonatal SD rat cardiomyocytes in PRKAG2 G100S group 48 h after infection. Conclusion Our findings indicate that myocardial glycogenosis might contribute to the pathogenesis of cardiac hypertrophy in patients with PRKAG2 G100S mutation, and the imbalance of calcium homeostasis in cardiomyocytes might not be involved in the pathogenesis.

Abstract:ObjectiveTo compare the safety and efficacy of ibutilide and amiodarone in converting post-coronary artery bypass grafting (CABG) new-onset atrial fibrillation (AF) to sinus rhythm. MethodsThis study was a prospective, single-blinded, randomized clinical trial. Totally 100 patients who met the inclusion criteria were enrolled and were intravenously treated with ibutilide (group A) or amiodarone (group B). In group A, Ibutilide was administered at a dose of 1 mg in 10 min, which was repeated after an interval of another 10 min if the arrhythmia was not converted; ibutilide was not given anymore no matter the conversion was achieved or not. In group B, amiodarone was administered at a dose of 5 mg/kg for the first 30 min, followed by 1 200 mg in the next 24 h; amiodarone was not given anymore no matter the conversion was achieved or not. Immediate conversion rate, conversion time and post-operative recurrence rate within 30 days were compared between the two groups. MethodsNo severe adverse reactions were observed in both groups. The immediate conversion rate of group A was significantly higher than that of group B (82% vs 58%, P<0.01), and the conversion time was significantly shorter than that of group B (66 min vs 660 min, P<0.01). The post-operative recurrence rates were not significantly different between the two groups within 30 days after operation (7.3% vs 6.9%,P>0.05). MethodsIbutilide and amiodarone are both safe in converting post-CABG new-onset AF to sinus rhythm, and ibutilide is more effective, more rapid in functioning and with a higher efficacy.

Abstract:[Abstract]ObjectiveTo investigate the therapeutic effect of argon-helium cryoablation combined with sorafenib on hepatitis B-related advanced hepatocellular carcinoma (HCC)and to evaluate the role of microvessel density (MVD) in prognostic evaluation. MethodsA total of 102 patients with advanced HCC were randomly divided into two groups, with 50 receiving sorafenib plus cryoablation(group S&C) and 52 receiving only cryoablation (group C). The endpoint of the treatment was tumor progression or untolerable adverse reaction. Tumor tissues were obtained before treatment. MVD was evaluated by immunohistochemical analysis using CD34 antibody. The therapeutic effects were evaluated according to RECIST criterion every 4-6 weeks. The adverse events were observed in both groups; the therapeutic effect, overall survival time(OS), and time to progression(TTP) were compared between two groups. MethodsIn group S&C, complete response (CR) was achieved in 2 patients(4%), partial response(PR)in 9 (18%), and stable disease(SD) in 22(44%), with the disease control rate(DCR) being 66%; in group C, no CR, PR in 4(7.6%), and SD in 19(36.5%), with the DCR being 44.2% (P<0.05). The overall survival (OS) and the time to progression (TTP) were significantly longer in group S&C than in group C (12.5 months vs 8.6 months, 9.6 months vs 5.3 months; P<0.01). The mean MVD in group CR&PR (111/0.74 mm2)was significantly lower than that in group PD(339/0.74 mm2, P<0.01). Patients with lower MVD receiving sorafenib plus cryoablation had longer OS and TTP than those only receiving cryoablation. The OS and TTP in patients with higher MVD had no significant difference between different groups. MethodsSorafenib plus cryoablation is a safe and effective therapy for patients with advanced HCC; it can improve the OS and TTP of patients. Patients with higher MVD have a lower response to therapy and poor prognosis.

Abstract:[Abstract]ObjectiveTo observe the effects of enriched environmen(EE) on brain-derived neurotrophic factor (BDNF) in substantia nigra (SN) of senescence-accelerated prone mice, so as to explore the possible mechanism of EE in alleviating MPTP-induced damage and in protecting dopaminergic neurons in the SN. MethodsTotally 80 3-month old female SAMP8 mice were averagely assigned to EE and standard environment (SE) groups at random. After three months, the mice in each group were further divided into 2 subgroups at random: MPTP group and NS group (n=20). The MPTP groups received subcutaneous injection of MPTP, and the NS mice were treated with an equal volume of NS. At the seventh day, the mice were sacrificed for RT-PCR and immunohistochemistry staining. RT-PCR was used to examine BDNF mRNA expression and immunohistochemistry staining was used to examine BDNF-ir expression. MethodsCompared with SE+NS mice, EE+NS mice had significantly increased BDNF mRNA expression(P<0.01), and the number of BDNF-ir cell and COD values in SN were also significantly increased (P<0.01). Compared with SE+NS mice, SE+MPTP mice showed significantly decreased BDNF mRNA expression (P<0.001), and the number of BDNF-ir cells and the COD values in SN of mice were significantly decreased (P<0.01). Compared with SE+MPTP mice, EE+MPTP mice showed increased BDNF mRNA expression (P<0.01), and the number of BDNF-ir cells and the COD values in SN were significantly increased (P<0.01).MethodsEE can increase BDNF mRNA expression and BDNF-ir cell number in SN of SAMP8 mice, alleviating MPTP-induced SN damage.

Abstract:[Abstract]ObjectiveTo investigate the presence of neutralization antibodies in the sera of hepatitis C patients. MethodsA eukaryotic expression plasmid encoding carboxyl terminal-truncated HCV envelope protein 2 (E2) was transfected into human 293T cells. ELISA method was established to examine the anti-E2 antibodies in the sera of 32 hepatitis C patients. The full-length envelope protein expression plasmid was transfected into 293T cells and the reactivity of transfectant with anti-E2 IgG positive sera was analyzed by immunofluorescence. Five strains of HCV pseudotype particle (HCVpp), including H77 (1a genotype), Con-1 (1b genotype), J4 (1b genotype), J6 (2a genotype), and UNK3a (3a genotype), and two strains of cell cultured HCV (HCVcc) were used to assay the neutralization activity of 12 anti-E2 positive serum samples. MethodsELISA results showed that 26 of 32 serum samples were anti-E2 IgG positive, with the positive rate being 81.3%. The 12 serum samples positive for HCV RNA were all anti-E2 IgG, and the virus load was negatively correlated with the anti-E2 antibody level. The anti-E2 positive sera could neutralize the infections of five strains of HCVpp and two strains of HCVcc to various extents, and the neutralization activity was consistent with the anti-E2 antibody level. MethodsIt is suggested that HCV infection can induce protective humoral immune response, and cross-neutralization antibodies against HCV are present in sera of hepatitis C patients, which indicates a feasibility for developing vaccines that can induce broadly-reactive neutralizing antibodies.

Abstract:[Abstract]ObjectiveTo study the effects of Exenatide on JAK1/STAT1 expression in the islet tissues and apoptosis of pancreatic B-cell in type 2 diabetic rat model with insulin resistance. MethodsInsulin resistant type 2 diabetic rat model was induced by combined treatment with high-fat diet and low-dose streptozotocin. The model rats were treated with Exenatide (Ex), metformin (Met) or normal saline (NS); the serum glycosylated hemoglobin (HbA1c), fasting insulin (FINS),and fasting plasma glucose (FBG) were observed and the insulin sensitivity index was calculated to evaluate the therapeutic effects of Exenatide on insulin resistance. Western blotting analysis was performed to detect the protein expression of JAK1/STAT1 in the islet tissues. Annexin/PI staining assay was used to evaluate B-cell apoptosis induced by oxidative stress. MethodsEx group showed a significantly elevated ISI and a reduced HbA1c level compared with NS group (P<0.05). JAK1/STAT1 expression and H2O2-induced B-cell apoptosis rate were significantly lower in Ex group than in Met and NS groups (P <0.01). MethodsExenatide can improve insulin sensitivity and inhibit B-cell apoptosis, probably by regulating the expression of JAK1/STAT1.

Abstract:[Abstract]ObjectiveTo investigate the expression and activation of MMP-2, MMP-9 in cerebral tissues of cerebral ischemia-reperfusion rats and the effects of doxycycline on the cerebral edema. MethodsCerebral ischemia-reperfusion model was established by middle cerebral artery occlusion (MCAO). The rats were randomly divided into a sham-operated and 6 reperfusion groups according to different reperfusion periods (3 h, 6 h, 12 h, 24 h, 72 h, and 120 h groups). Dry weight method was used to evaluate hemispheric water content of the ischemic side after treatment with doxycycline. Western blotting analysis and Gelatin zymography were used to determine the expression and activity of MMP-2 and MMP-9 in the ischemic cerebral tissues. MethodsThe protein expression and activity of MMP-2 were significantly increased in the ischemia tissue at 3 h and 120 h after reperfusion(P<0.01); the protein expression and activity of MMP-9 began to increase at 6 h after reperfusion(P<0.01), reached the peak at 24 h, and then returned to the basal levels at 120 h after reperfusion (P>0.05). The hemispheric water content of the ischemic side gradually increased with reperfusion periods compared with the sham-operated group. Dexycycline significantly reduced the hemispheric water contents compared with the normal saline at the same time points(P<0.05 or P<0.01). MethodsMMP-2, MMP-9 can induce vasogenic brain edema by degrading basal membrane of capillary vessel. The alteration of MMP-2, MMP-9 expression and activities are likely associated with brain edema during cerebral ischemia-reperfusion.

Abstract:[Abstract]ObjectiveTo explore the diagnostic value of serum Cys-C, hs-CRP, and urine mALB for early renal injury in gonty patients.MethodsA total of 100 gonty patients and 50 healthy volunteers were divided into 3 groups: acute phase group (n=50), intermittent group (n=50), and healthy control group (n=50) . The serum Cys C,hs-CRP,and urine creatinine were determined by automatic chemistry analyzer and urine mALB by immunofluorometer equipment, and the results were analyzed in each group.MethodsThe results of mALB/Cr(mg/\[g·Cr\]), hs-CRP(mg/L), and Cys-C(mg/L)in acute phase group were 11.45±3.55, 5.15±1.98, and 2.62±0.83, in intermittent group were 8.96±1.78, 3.28±1.23, and 0.93±0.26, and in healthy volunteer group were 3.89±0.76, 1.91±0.57, and 0.78±0.22, respectively. The values of the above three parameters in the acute phase and intermittent groups were significantly higher than those in the healthy control group (P<0.05); and the values in the acute phase group were significantly higher than those in the intermittent group (P<0.05).The abnormal rate of hs-CRP+Cys-C+mALB/Cr was 87% in gonty patients.MethodsSimultaneous determination of mALB/Cr, hs-CRP, and Cys-C can help to assess the early renal injury in gonty patients.

Abstract:\[Abstract\]ObjectiveTo discuss the technique of the anterior cervical decompression via intervertebral space under microscope. MethodsA total of 271 patients with cervical disc herniation underwent anterior microsurgical decompression and intervertebral fusion from Jun. 2000.6 to Dec. 2005. The patients included 178 cases with single level herniation, 71 with double level ones, and 22 with three level ones. MRI images showed slight or medium anterior compression with slight abnormal changes within spinal cord in all cases. The main equipment included operation microscope for neurosurgery and high speed bur. The annular fibrous and osteophytes were removed by high speed bur to expose and clean the degenerative nucleus pulpous. We lifted the posterior longitudinal ligament (PLL) with tension using a hook-like stripper and cut it; then the residual PLL and posterior bony edges were removed. MethodsThere were no severe complications such as injuries of the spinal cord, nerve root or blood vessels. The average intraoperative blood loss was (126±29.4) ml and the mean operative time was (62±12.4) min. The average time for the patients to get off bed was (1.8±0.4) days. The nerve functions were improved in all patients after operation, with the excellent nerve function found in 69 cases, good in 157 cases, fair in 27 cases, and poor in 18 cases according to JOA scores. MethodsAnterior cervical decompression via intervertebral space under microscope is suitable for treatment of pathological changes of the intervertebral space; delicate manipulation under microscope and high speed bur can contribute to a successful operation; and the anatomy of intervertebral space is more suitable for the biomechanics of the cervical vertebrate.

Abstract:[Abstract]ObjectiveTo evaluate whether combined supplementation of vitamin C and E in pregnant women can reduce the risk for preeclampsia. MethodsElectronic databases were searched for randomized controlled trials investigating pregnancy outcomes in women following exposure to combined vitamin C and E supplementation since the establishment of the database to Oct. 2010. The end-point of the study was preeclampsia. The relative risk (RR) and confidence interval (CI) for each included studies were pooled and heterogeneity analysis was performed by using RevMan 5.0.25 software. MethodsTotally 10 studies involving 19 656 pregnant women at risk of preeclampsia were included, with 9 823 receiving vitamin C and E and 9 833 receiving placebo. The pooled RR values for preeclampsia in all studies, the studies with high quality, and the studies with low quality were 0.94 (95% CI of 0.80-1.11, P=0.49), 1.02 (95% CI of 0.91-1.14, P=0.79), and 0.40 (95% CI of 0.15-1.10, P=0.08), respectively. MethodsCombined vitamin C and E supplementation has no potential benefit in reducing the risk for preeclampsia.

Abstract:[Abstract]ObjectiveTo study the influence of Salvia miltiorrhiza bge.f.alba(SMA) on apoptosis and Bcl-2 expression in human umbilical vein endothelial cell (HUVEC) in vitro.MethodsHUVECs were isolated using perfusion and enzyme digestion methods, and the obtained cells were identified by morphological observation and ⅧAg immunoreactivity examination. The cells in the exponential phase of growth were treated with H2O2 and different concentrations of SMA(high dose: 0.10 g/ml, low dose: 0.01 g/ml). The cell apoptosis was determined by flow cytometric analysis and Bcl-2 expression was examined by immunofluorescence mehtod. MethodsSMA significantly decreased the H2O2-induced apoptosis of HUVECs (P<0.01 in high dose group and P<0.05 in low dose group), and significantly increased the expression of Bcl-2 in high dose group(P<0.01). MethodsSMA can inhibit H2O2-induced apoptosis of HUVEC, which might be associated with the increase of Bcl-2 expression.

Abstract:[Abstract]ObjectiveTo establish a multivariate reference range for ECG while considering the relation between multiple variables. MethodsA total of 777 female undergraduates in a university were taken as the subjects and the P-wave amplitude was taken as an example to establish a multivariate reference range for P-wave amplitude by using Mahalanobis Depth Function. MethodsP-wave amplitude of the female undergraduates was established as follows: for a 95% CI of multivariate reference range,Depth≥0.035 521 6. MethodsMultivariate reference range for ECG obtained in this study allows for a comprehensive utilization of the ECG data and can help the clinical diagnosis using ECG.

Abstract:[Abstract] Objective To optimize the extraction process for phenolic glycosides from Rhizoma Curculigins. Methods The contents of orcinol glucoside,curculigoside and curculigine were used as parameters, and they were determined by HPLC. The extraction conditions of phenolic glycosides from Rhizoma Curculigins were optimized using orthogonal design. The examined factors included concentration of ethanol(50%, 70%, and 95%), extraction time (1, 1.5, and 2 h), times of extraction (1, 2, and 3), and solvent volume (6, 8, and 10 times).Methods The concentration of ethanol, extraction time, times of extraction and solvent volume significantly affected the extraction efficacy of phenolic glycosides in Rhizoma Curculigins, with the most prominent effect exerted by the ethanol concentration. The optimal extraction process was refluxing three times with 70% ethanol (10 times of solvent volume), for 2 h each time. Methods The present method can effectively extract phenolic glycosides from Rhizoma Curculigin, which paves a way for further research on phenolic glycosides in Rhizoma Curculigins.

Abstract:[Abstract]ObjectiveTo study the effect of training in the humid heat environment on motion sickness by systemically training volunteers in the humid head environment. MethodsSixty volunteers, aged 18-23 years old, were selected from 567 male college students. All the subjects passed health examination and the Soldier Physical Ability Standard test of Chinese People’s Libration Army. They were equally divided into an experimental group and a control group. The experimental group received adaptation training with motion sickness device in humid heat environment (35-39℃, 60%-85% humidity). Motion sickness tests were done in both groups before and after training. The Criolis acceleration rotational chair was used to induce the motion sickness and Graybiel scale was used to evaluate the degree of motion sickness. MethodsThe degrees of discomfort were defined as following: N, no; Ⅰ, slight; ⅡB, moderate B; ⅡA, moderate A; Ⅲ, severe; and F, serious. The motion sickness degree distribution in the experiment group was (1 dropout due to sickness) N, 2; Ⅰ, 0; ⅡB, 0; ⅡA, 0; Ⅲ, 21; F, 6 before the training; and N, 0; Ⅰ, 3; ⅡB, 7; ⅡA, 4; Ⅲ, 14; F, 1 after the training. The distribution of the control group was N, 1; Ⅰ, 1; ⅡB, 0; ⅡA, 1; Ⅲ, 22; F, 4 before the training; and N, 0; Ⅰ, 0; ⅡB, 5; ⅡA, 3; Ⅲ, 16, F, 5 after the training. MethodsTraining in humid heat enviroment can achieve a great decrease in motion sickness degree, with the average motion sickness degree in the experiment group greatly improved compared with that in the control group.

Abstract:[Abstract]ObjectiveTo report the first case of Churg-Strauss syndrome (CSS) manifested as IgA nephropathy in China. MethodsThe clinicopathological data for diagnosis of the patient were analyzed, and the related literatures were reviewed. MethodsThe female patient aged 39 years old. Renal needle biopsy was performed 6 years ago because of gross hematuria and proteinuria, with the diagnosis being IgA nephropathy and focal segmental glomerulosclerosis. Laboratory examinations showed that the proportion of peripheral eosinophils was increased and c-ANCA/p-ANCA was positive once. Sufficient prednisone and cyclophosphamide were given for pulse therapy for 18 months, and the patient was clinically cured. One year ago, the patient received peritoneal dialysis because the serum creatinine reached 932 μmol/L. However, the general status of the patient was not improved after dialysis, and the patients developed cough, diarrhea and low-grade fever. Repeated blood test showed that the proportion of peripheral eosinophils was continuously increased. p-ANCA and MPO-ANCA were positive, and CT scan indicated chronic sinusitis. The patient had a history of “rashes”, therefore CSS was diagnosed. The patient was given pulse therapy with immunoglobulin treatment for 3 days and oral prednisone at a daily dose of 30 mg; the temperature of the patient decreased immediately to the normal level; and her general condition was improved and she was discharged. Methods IgA nephropathy was a rare pathological manifestation of CSS, which should be considered during diagnosis.

Abstract:[Abstract]Objective To understand the risk of heterotopic pregnancy when intrauterine pregnancy and abdominal pain co-exist. Methods and resultsWe report a patient with heterotopic pregnancy in a naturally occurring pregnancy; she also had high risks for secondary infertility. At gestational week 13 she had hemorrhagic shock due to intrauterine pregnancy complicated with rupture of tubal pregnancy; salpingectomy was performed via open operation. The intrauterine pregnancy was uneventful after operation, and a healthy baby was delivered through cesarean section at 37+2 weeks due to placental previa. MethodsHeterotopic pregnancy usually occurs in women receiving assisted reproductive technology, but it can also happen in natural conception when there are high risk factors. Satisfactory outcome can be obtained in patients with heterotopic pregnancy, rupture of tubal pregnancy, and hemorrhagic shock, if prompt measures and supporting treatment can be given.

Abstract:\[Abstract\]Nano-materials have been increasingly widely used in various fields with the progression in science and technology. While bringing benefit to our life, their toxic effects also come to our attention. In this paper we review the cytotoxic effects and some problems of several kinds of nano-materials (carbon-based nano-materials, metal and metallic oxides and other nano-materials), considering the physicochemical characteristics of nano-materials, research methods of the nano-material cytotoxic effects, and the cytotoxcity of nano-materials. Finally, we also discuss the research focus of the area and issues need to be addressed, hoping to provide a guide for bio-safety research of nano-materials.

Abstract:\[Abstract\]Candida albicans, the most common fungal pathogen, can cause mucosal and systemic infections. Until now there are only a limited number of antifungal drugs, and long-term repeated use of the existing drugs has led to rapid development of drug resistance. The high adaptability of C. albicans has led to drug resistance, which is also a manifestation of environmental stress of C. albicans to the environment. This article summarizes the response pathways of C. albicans stress and describes drug resistance of C. albicans from the aspect of stress response.

Abstract:[Abstract]ObjectiveTo optimize the extraction process for phenolic glycosides from Rhizoma Curculigins. MethodsThe contents of orcinol glucoside,curculigoside and curculigine were used as parameters, and they were determined by HPLC. The extraction conditions of phenolic glycosides from Rhizoma Curculigins were optimized using orthogonal design. The examined factors included concentration of ethanol(50%, 70%, and 95%), extraction time (1, 1.5, and 2 h), times of extraction (1, 2, and 3), and solvent volume (6, 8, and 10 times).MethodsThe concentration of ethanol, extraction time, times of extraction and solvent volume significantly affected the extraction efficacy of phenolic glycosides in Rhizoma Curculigins, with the most prominent effect exerted by the ethanol concentration. The optimal extraction process was refluxing three times with 70% ethanol (10 times of solvent volume), for 2 h each time. MethodsThe present method can effectively extract phenolic glycosides from Rhizoma Curculigin, which paves a way for further research on phenolic glycosides in Rhizoma Curculigins.

Abstract:[摘要]目的观察人脂肪组织来源的神经干细胞移植对大鼠局灶性脑缺血后VEGF表达的影响，探讨其治疗局灶性脑缺血的可能机制。方法线栓法制作大鼠大脑中动脉缺血(MCAO) 2 h再灌注模型。60只健康雄性SD大鼠随机分为4组：正常对照组(6只)，假手术组(6只)，缺血对照组(24只)和移植治疗组(24只)。后两组又各分4组：缺血2 h再灌注7、14、21、28 d组(各6只)。体外培养脂肪基质细胞，诱导分化为神经干细胞。造模成功后24 h，移植治疗组经尾静脉移植人脂肪组织来源的神经干细胞悬液(细胞密度为2×106/ml)，缺血对照组经尾静脉注射生理盐水，假手术组不做任何处理。免疫组化法检测VEGF的表达，原位杂交法检测VEGF mRNA的表达。结果与缺血对照组比较，移植治疗组各时间点(7、14、21、28 d)的VEGF染色阳性细胞数均增加(均P<0.01)，VEGF mRNA含量均增加(均P<0.01)。结论脂肪组织来源的神经干细胞移植可促进内源性VEGF及VEGF mRNA的表达，提高大鼠局灶性脑缺血后损伤局部VEGF及VEGF mRNA的含量，对脑缺血再灌注损伤后的神经细胞起保护作用。

Abstract:[摘要]目的获得具有较高纯度和酶活性的烟酰胺单核苷酸腺苷转移酶-1(Nmnat1),为进一步对其功能及调控的研究奠定基础。方法在大肠杆菌中进行Nmnat1表达的条件优化,采用Ni-NTA 亲和层析法对目的蛋白进行纯化,并通过酶联荧光法测定酶活性。结果BL21-CondonPlus (DE3)-RIL为适宜的宿主菌,优化的蛋白表达条件为: 在2×YT培养基(37 μg/ml氯霉素和75 μg/ml卡那霉素)中于28℃、0.5 mmol/L IPTG(isopropyl beta-D-thiogalactopyranoside, 异丙基-β-D-硫代半乳糖苷)诱导8 h。纯化获得的Nmnat1蛋白具有较高的纯度和酶活性。结论适宜的宿主菌对Nmnat1蛋白的高效表达至关重要,进行表达条件优化后可获得大量具有较高纯度和酶活性的目的蛋白。

Abstract:[摘要]目的观察低浓度三氧化二砷(As2O3)在不同氧环境下对肺癌A549细胞增殖抑制、细胞周期阻滞、细胞凋亡的诱导作用。方法分别在常氧(21% O2)、低氧(5% O2)、厌氧(0% O2)环境利用0 μmol/L As2O3(空白对照组)、1 μmol/L As2O3、1 μmol/L VP-16体外诱导培养肺癌A549细胞24 h,通过甲基噻唑基四唑(methyl thiazolyl tetrazolium, MTT)法测细胞增殖抑制率,通过碘化丙啶(propidium iodide, PI)染色、流式细胞仪检测细胞周期,通过Annexin Ⅴ/PI双染色法检测细胞凋亡。结果1 μmol/L As2O3组A549细胞增殖抑制率及G0/G1期细胞比例随缺氧加重而增加；厌氧环境下该组细胞增殖抑制率及G0/G1期细胞比例均高于空白对照组(P<0.05),但其细胞凋亡率较空白对照组降低(P<0.05)；与3种相应氧环境下VP-16组相比较,As2O3组A549细胞增殖抑制率、G0/G1期细胞比例、G2/M期细胞比例、细胞凋亡率均减少(P<0.05)。结论1 μmol/L As2O3对A549细胞无明显诱导凋亡作用,厌氧环境下该剂量As2O3可以通过G1期阻滞抑制A549细胞增殖。

Abstract:[摘要]目的探讨大鼠脑缺血再灌注损伤对血脑屏障内皮屏障抗原(EBA)及通透性的影响。方法20只雄性SD大鼠随机分成为缺血再灌注组和假手术组,采用阻断大鼠一侧大脑中动脉(MCA)血流2 h、再灌注24 h制成局灶性脑缺血模型。24 h后断头取脑,制作冰冻切片,免疫组化染色检测血脑屏障EBA和Fibrinogen的表达。结果缺血再灌注组EBA免疫反应阳性血管数5.30±1.24,假手术组EBA免疫反应阳性血管数15.20±1.02,两组比较差异有统计学意义(P<0.01)。缺血再灌注组Fibrinogen免疫反应阳性血管数8.60±3.12,假手术组Fibrinogen免疫反应阳性血管数1.30±0.46,两组比较差异有统计学意义(P<0.01)。结论脑缺血再灌注损伤能显著减少血脑屏障EBA表达,增加血脑屏障通透性。

Abstract:[摘要]目的检测二甲双胍作用下肝细胞癌SMMC-7721细胞的增殖和凋亡情况,并初步探讨了其中的机制。方法以不同浓度(0.2、1、2、5 mmol/L)二甲双胍处理SMMC-7721,用CCK-8法检测其对细胞增殖的影响,Hochest33342 DNA染色检测细胞凋亡,实时定量PCR法检测Bcl-2和Bid mRNA的表达情况。结果二甲双胍处理48 h 和96 h后,人肝细胞癌SMMC7721细胞的增殖明显受到抑制(P<0.05),以2 mmol/L和5 mmol/L组最为明显(P<0.01),并呈时间和剂量依赖性。Hochest33342染色发现,1 mmol/L和5 mmol/L浓度的二甲双胍可促进细胞凋亡,细胞凋亡率与对照组相比差异有统计学意义(P<0.05)。二甲双胍处理后,SMMC7721细胞内抗凋亡分子Bcl-2表达下降(P<0.01),促凋亡分子Bid表达升高(P<0.05)。结论二甲双胍能抑制人肝细胞癌SMMC7721细胞增殖,促进细胞凋亡,其机制可能与细胞内抗细胞凋亡分子Bcl-2表达下降及促凋亡分子Bid表达升高有关。

Abstract:[摘要]目的采用蛋白质组学方法分析、寻找后纵韧带骨化症(OPLL)的特异性蛋白标志。方法采用荧光差异双向电泳法研究OPLL患者及正常人韧带组织的双向电泳图像。采用飞行时间质谱进行鉴定并分析差异表达的蛋白质点。结果两组样本均得到约1 100个蛋白质点,50个点表达有改变。切胶获得45 个蛋白质点的肽段样本,质谱鉴定为21个蛋白质或肽段。其中15个血液蛋白,其余6个蛋白中4个表达下调；2个表达上调。RT-PCR检测表明烟酰胺腺嘌呤二核苷酸依赖的激素脱氢酶, 伴肌动蛋白相关锚定蛋白及Ⅵ型胶原蛋白mRNA变化与电泳结果一致。结论这些差异表达蛋白可能是OPLL的特异表达蛋白。

Abstract:[摘要]目的比较蛛网膜下隙阻滞与全身麻醉应用于老年患者髋关节置换术中的疗效,为临床选择合适的麻醉方法提供依据。方法根据电子麻醉记录单回顾341例分别在蛛网膜下隙阻滞和全身麻醉下行髋关节置换的老年患者术中血压变化、出血量和输液输血量、手术时间和术后恢复时间等。结果蛛网膜下隙阻滞组血压变化较全身麻醉组小,失血量少,术后恢复时间短(P＜0.05)。结论蛛网膜下隙阻滞和全身麻醉均可安全有效地用于老年人髋关节置换术,但蛛网膜下隙阻滞在对循环的影响和恢复速度上要优于全身麻醉。

Abstract:目的比较3种气道控制方式对麻醉患者血流动力学及听觉电位诱发指数(AAI)的影响。方法选择无困难气管插管和重要脏器疾患ASA Ⅰ~Ⅱ级,择期全麻手术患者90例,随机分成3组：A组(直视下气管插管组)、B组(光棒引导下气管插管组)和C组(喉罩放置组),每组30例。记录各组患者入手术室时(基础值T0)、气管插管/喉罩放置前(T1)、气管插管/喉罩放置即刻(T2)的AAI、脉搏氧饱和度(SpO2)、心率(HR)、收缩压(SBP)、舒张压(DBP),计算基础值与气管插管/喉罩置入前的差值(T1-0)、基础值与气管插管/喉罩置入即刻的差值(T2-0)。 结果3组操作前AAI值均明显下降,表明达到适当的麻醉深度。3组操作前血压下降、心率增快的作用均有,A组直视下气管插管操作明显升高血压、增快心率,与B组和C组比较差异具有统计学意义(P<0.05)。B组和C组操作时血压和心率没有明显差异,但AAI值有明显差异,B组AAI值升高程度明显高于C组(P<0.05)；操作引起AAI值升高的顺序依次为A、B、C组。结论光棒引导下气管插管与放置喉罩对患者心血管系统影响均小,但喉罩组对AAI的刺激作用更小。

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