Characteristics of Ex Vivo Expansion of Endothelial Progenitor Cells

Recently, endothelial progenitor cells (EPCs) were iso- lated from peripheral blood and proved to participate in neovascular formation[1-3]. This will be useful in regener- ating therapeutic vasculogenesis, provide suitable coat- ing for vascular grafts or deliver lethal toxins to tumor vascular beds. But EPCs in peripheral blood is very lim- ited in number, its introduction into transplantation pro- tocols has been hampered by the limited number of pro- genitors contained in blood harvested…     

Role of Caspase-3 Inhibitor in Induced Anoikis of Mesenchymal Stem Cells In Vitro

By preventing mesenchymal stem cells (MSCs) from adhering to precoated agarose to create a model of MSC suspension in vitro, we investigated anoikis in MSCs and the role of cas- pase-3 in the anoikis. The cultured MSCs were randomly divided into 3 groups: the anoikis group, caspase-3 inhibitor group and control group. Before experiment, we coated dishes with 1.5 % agarose; in the anoikis group, MSCs were put into the precoated dishes; and in the inhibitor group, caspase-3 inhibitor and MSCs were also put into the precoated dishes; but there were not intervention in the control group. MSCs were collected at 2 h, 6 h, 12 h and 24 h. The alteration of caspase-3 activity was evaluated by caspase-3 fluorometric assay and western blot analysis. The apoptosis rates were detected by flow cytometry. MSCs were round and suspended sufficiently in the anoikis and inhibitor groups. Caspase-3 fluorometric assay showed that there were significant differences in statistics be- tween the anoikis group and the others (P<0.05). Western blot analysis discovered that caspase-3 ex- pression in the anoikis group was more than that in the control and inhibitor groups (P<0.05). Flow cytometry showed that the apoptosis peak appeared in all the three groups, but it increased dramati- cally in the anoikis group. The apoptosis rates in the inhibitor and control groups were low and stable. And there were significant differences in statistics between the anoikis group and the others (P<0.05). MSCs will undergo anoikis in suspended condition if they are separated from the extracellular matrix. Caspase-3 inhibitors can suppress caspase-3 activity and reduce the apoptosis rate significantly. Cas- pase-3 plays a vital part in induced MSC anoikis in vitro. MSCs suspension culture system might be set up with argorose and caspase-3 inhibitor.     

A Study of Tumor Apoptosis Induced by Medicine Serum of Traditional Chinese Medicine

Objective Adopting serological methods of compound Chinese medicine that including ＇Yi-Fei-Yin＇ to treat lung cancer, ＇Yi-Chang-Tai＇ to treat lung cancer and ＇Wei-Ai-Ning＇ to treat gastric cancer to explore the induced effect on the tumor cell apoptosis. Methods （1） Culture conventionally A549 human lung cancer cell lines, CCL229 human colorectal carcinoma cell lines and gastric cancer cell line SGC7901, and observe under the inverted microscope. （2） Three Chinese medicine compound which were decoction, three consecutive days of intragastric administration to rats, last administration was two hours before the animals were killed, according to the serum drug Preparation prepared medicinal serum. （3） Three drugs were cultured in serum of A549 human lung cancer cell lines, CCL229 human colorectal carcinoma cell lines and cell line SGC7901 48 hours. （4） With rubber scraper scraped cells, cell suspension will be transferred to the centrifuge tube centrifuge 2000 rpm/min, 15min, Agar processing into pieces, washed with PBS, 1% osmium acid fixed, gradient ethanol and acetone dehydration, epoxy-embedded mesh, the conventional ultra thin section, electron stain. （5） To observe and take a picture under the JEM1200 TEM. Results （1） The parapodium of tumor cells decreased even disappeared, endocytoplasmic reticulum, mitochondria and other cell organelles for Poly IC, chromatin condensation, margination a ＇crescent＇. （2） It is apoptosis characteristic structure--the membrane parcels, containing some of the broken device and cell chromatin of apoptotic bodies. Conclusion Compound Chinese medicine serum has the function to induce tumor cells apoptosis, widely used for clinical medicine compound against lung cancer, gastric cancer and colorectal cancer provide reliable experimental evidence.     

The Effect of HBx Gene on the Apoptosis of Hepatic Cells

To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the mor- phology of L02/pcDNA3.1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/ pcDNA3.1-HBx; (3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P<0.01) and the apoptosis rate of HepG2/pcDNA3.1-HBx was lower than that of HepG2 cells (P<0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P<0.01), and the expression of XIAP in HepG2/pcDNA3.1-HBx was about 4 times that in HepG2 (P<0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of he- patic carcinoma by regulating the expression of XIAP.     

Study of apoptosis pattern of dopaminergic neurons and neuroprotective effect of nicotine in MPTP mouse model

Objective：To investigate the apoptosis of dopaminergic neurons and the protective effect of nicotine in 1-methyl-4-phenyl- 1, 2,3,6-tetrahydropyridine （MPTP）-induced mouse model of Parkinson＇s disease. Methods：The mouse model of Parkinson＇s disease were formed by MPTP （30 mg/kg/d×7, i.p. ）; and the loss and apoptosis of dopaminergic neurons was observed by Tyrosine Hydroxylase（TH） and TUNEL stains.In ＂Nicotime plus MPTP＂ group, mice were pretreated with nicotine before MPTP injection. The putative protective effect of nicotine was analyzed. Results：The number of TH-positive cells decreased during MPTP treatment. Apoptotic neurons began to appear after three injections of MPTP and peaked on the 8th day. In the MPTP-intoxicated mice treated with nicotine, the loss of TH-positive cells was significantly less than that of MPTP- treated group （30 mg/kg/d×7）（P 〈 0.05）. Conclusion：The chronic treatment of MPTP can induce the apoptosis of dopaminergic neurons in substantia nigra, and nicotine might have a neuroprotecitve effect on dopaminergic neurons against MPTP toxicity.     

Effects of the Extract of Ammopiptanthus mongolicus cheng f. （JA1） on Induction of Apoptosis of HepG2 in vitro and Its Molecular Mechanisms

Objective To study the effects and the mechanisms of extract from a leguminous plant （Ammopiptanthus mongolicus cheng f.） （JAl ） in northwest China on inducing apoptosis and inhibiting proliferation of HepG2 hepatocarcinoma cell in vitro. Methods The HepG2 cell line was used as target cells. The effect of 3A 1 on HepG2 cell growth was detected by microculture tetrazolium assay （MTr）, flow cytometry assay, DNA agarose gel electrophoresis and transmission electronic microscopy. The expressive effect of the wt-p53 in HepG2 cells was analyzed with p53 protein test-reagent. Results JAl not only had significant anti-proliferative effects depending upon time and dosage, but also induced apoptosis of HepG2 cells. Apoptotic typical morphological changes were observed in JAl-treated HepG2 cells under transmission electronic microscope, ＂Sub-G 1＂ phase peak occurred in flow cytometry and DNA ＂ladder＂ was found in DNA agarose gel electrophoresis. The expression of the wt-p53 increased in vitro, and 3Al-treated HepG2 and the positive cell percentage of the wt-p53 protein also increased. Conclusions JAl could obviously induce apoptosis and inhibit proliferation of HepG2 cells in vitro, and these effects are closely related with the increase of wt-p53 expression. JAl can be used as a good source of medicinal plant for the treatment of hepatocarcinoma.     

Association between Up-regulation of Fas Ligand Expression and Apoptosis of Tumor-infiltrating Lymphocytes in Human Breast Cancer

In order to study the significance of FasL expression in immune escape of breast cancer, FasL protein expression and the number of tumor-infiltrating lymphocytes (TILs) in 40 specimens of breast cancer were detected by immunohistochemitry. The expression of FasL mRNA was measured by in situ hybridization in the consecutive tissue slices of 40 breast cancers respectively. By using terminal deoxynucleotidyl transferase-mediaed dUTP nick end labeling (TUNEL), apoptotic cells were detected in 40 specimens of breast cancer. The expression of FasL was detected in all 40 specimens to varying degrees. In the consecutive tissue slices, the location of expression of FasL protein corresponded with that of FasL mRNA. In those with FasL extensive expression, the number of TILs was less (P<0.05), the apoptotic index (AI) of TILs was higher and the AI of tumor cells was lower (P<0.01) than those with FasL weak expression respectively. The AI of TILs was correlated with that of tumor cells (r=-0.629, P<0.01). In conclusion, breast cancer cells can induce the apoptosis of TILs through the expression of FasL, which can counterattack the immune, system. This may be a mechanism of immune evasion in breast cancer.     

Experimental study on inhibitory effect of bridgy duct of hepatic artery on apoptosis of liver and bile duct cells

Objective： To investigate whether the method of bridgy duct established between the recipient＇s spleen artery and the donor＇s gastroduodenal artery could inhibit the apoptosis of liver and bile duct cells caused by hepatic artery ischemia. Methods： Twenty-four mongrel dogs from Xi＇an area were used to establish simplified models of dog orthotopic liver transplantation and divided into three groups randomly： HAI group （hepatic artery ischemia group）,BBB group（bypassing the blood by a bridgy duct） and control group. After cold perfusion, The samples were collected from liver and bile duct in each group at different time and fixed in glutaraldehyde and 4% polyformaldehyde respectively. At last, the apoptosis of liver and bile duct cells were observed and the apoptotic indexes were calculated. Results： Two hours after cold perfusion, apoptotic phenomenon was common in HAI group, rare in BBB group, while no apoptotic phenomenon was observed in control group. TUNEL staining showed that there was no significant difference in apoptotic index among the three groups immediately after cold perfusion. However, with time going, the apoptotic cells were increased in three groups, and the difference in apoptotic index was significant among three groups （P 〈 0.01 ）. Conclusions： Bridgy duct of hepatic artery can inhibit the apoptosis of liver and bile duct cells caused by HAI significantly.     

Quantitative morphometry and apotosis of colorectal carcinoma

目的：探讨大肠疾病形态计量学和凋亡规律。方法：应用图像自动分析系统，检测了大肠疾病形态计量学参数和凋亡指数（AI）。结果：大肠腺癌AI明显高于大肠腺瘤和炎性增生组（P分别＜O．05和＜O．001）；细胞核面积、周长、凸状周长、等效直径、周长面积因子和核平均灰度六项参数，在腺癌组明显高于腺瘤和炎性增生，而灰度标准差明显低于其他两组，大肠腺癌AI与PM2／A相关系数为O．51（P＜0．05）。结论：形态计量学参数和凋亡指数在鉴别大肠疾病中具有一定参考价值。     

Changes of Apoptosis in Rats of Acute Ischemic Renal Injury under Treatment of Tetrandrine

Objective To elucidate the effect of tetrandrine on acute ischemic renal injury and its relation with apoptosis.Methods A model for bilateral post-ischemic renal injury in rats was developed by clamping renal pedicles for 45 min.Renal tissular DNA fragmentation analysis and renal tissular HE staining were used.Also quantitative analysis of apoptosis in injured renal tubular epithelium was carried out by using TdT-mediated dUTP nick and labeling(TUNEL).Results Apoptosis of renal tubular epithelium increased in acute ischemic renal injury.Tetrandrine could remarkably decrease the level of apoptosis in injured renal tubule while protecting renal tissue against the ischemic injuries.Conclusion Tetrandrine could adjust the level of apoptosis in renal tubular epithelium and alleviate renal tissular injury.     

Clinical study on Tongyan Spray (通咽喷雾剂) for post-stroke dysphagia patients: A randomized controlled trial

Objective:To observe the effectiveness and safety of Tongyan Spray(estern Medicine,Beijing(100039),China;Jiedu Granules;Cinobufacini Injection which was extracted from skin of Bufo     

MicroRNA expression patterns of the kidney in hyperuricemia mice treated with Xiezhuo Chubi Decoction (泄浊除痹方)

Objective:To investigate the effects of Xiezhuo Chubi Decoction(泄浊除痹方,XZCBD) on the microRNA expression patterns of kidney in mice with hyperuricemia.Methods:Sixty Kunming male mice were randomly divided into the high-,medium-,and low-dose XZCBD groups,benzbromarone group,model group, and control group.Except the control group,all mice were established with yeast method combined with uricase inhibition method to build hyperuricemia model,and the corresponding drugs(37.5 g/kg,18.75 g/kg,9.375 g/kg, and 0....     

Clinical Observation on the Treatment of Chronic Severe Hepatitis B by Retention Enema with Huchang Jiedu Decoction(护肠解毒汤)

<正>Objective:To observe the efficacy of retention enema with Huchang Jiedu Decoction(护肠解毒汤, HJD)in treating chronic severe hepatitis B(CSHB).Methods:Sixty patients of CSHB were equally randomized into the treated group and the control group.Both groups were treated with conventional integrative medicine, but to patients in the treated group,retention enema with HJD was given in addition,once every day for 3 weeks.The dominant symptoms,physical signs,and related biochemical indices,as well as the incidence of complications in patients before and after treatment,were observed.Results:Good therapeutic effects were shown in the treated group,with a total effective rate better than that in the control group(83.3%versus 60.0%, P0.05),superior in terms of lowering alanine aminotransferase(ALT),aspartate aminotransferase(AST), total bilirubin(TBil),globulin(GIb),and endotoxin(ET)levels and increasing prothrombin activity(PTA),total cholesterol(TC),and calcium(Ca)levels,as well as eliminating ascites and preventing hepatic encephalopathy (P0.05);especially in treating middle/early stage patients with Chinese medicine syndrome differentiated as water-toxin accumulation pattern.Conclusion:Retention enema with HJD is surely effective in treating CSHB, and its primary mechanism may be related to the mitigation of enterogenous endotoxemia.     

Anti-fatigue effect of Renshen Yangrong decoction (人参养荣汤) in mice

Objective  

To explore the anti-fatigue effect of Renshen Yangrong Decoction (人参养荣汤, RYD): in mice.     

A randomized controlled single-blind clinical trial on 84 outpatients with psoriasis vulgaris by auricular therapy combined with optimized Yinxieling Formula (银屑灵优化方)

Objective:To explore the therapeutic effect of auricular therapy combined with optimized Yinxieling Formula(银屑灵优化方) on psoriasis vulgaris.Methods:A randomized controlled single-blind clinical trial on 84 outpatients with psoriasis vulgaris was conducted.The patients were randomized to a treatment group(43 cases treated by auricular therapy combined with optimized Yinxieling Formula) and a control group (41 cases treated by optimized Yinxieling Formula alone) according to a random number generated by SPSS 17.0 software.The treatment duration for both groups was 8 weeks.The therapeutic effect was comprehensively measured by the primary outcome measure[Psoriasis Area and Severity Index(PASI) reduction rate]and the secondary outcome measure[PASI,Visual Analogue Scale(VAS),Dermatology Life Quality Index(DLQI), Self-rating Depression Scale(SDS),and Self-rating Anxiety Scale(SAS)].The outcomes of both groups were obtained and compared before and after the intervention.Results:The PASI reduction rate in the treatment group was 74.4%(32/43),which was higher than that in the control group(36.6%,15/41,P<0.01).The PASI scores decreased in both groups after treatment and was lower in the treatment group compared with the control group (P<0.01).With stratified analysis,there were significant differences between the PASI scores in the following subgroups:age 18-30,baseline PASI>10 and stable stage(P<0.05).DLQI decreased in both groups on some categories after treatment,but there were no significant differences between the two groups in SDS,SAS and VAS(P>0.05).No obvious adverse reactions were found in either group.Conclusion:The therapeutic effect of auricular therapy combined with Optimized Yinxieling Formula was superior to Optimized Yinxieling Formula alone with no obvious adverse reaction.     

Effect of Qingre Quyu Granule (清热祛瘀颗粒) on stabilizing plaques in the brachiocephalic artery of apolipoprotein E deficient mice

Objective： To investigate the effect of Qingre Quyu Granule （清热祛瘀颗粒, QRQYG） on stabilizing vulnerable plaques in apolipoprotein E （ApoE） defficient mice. Methods： Seventy-two male ApoE defficient mice were given a high-fat diet from 6 weeks of age. At the 16th week, all the mice were randomized into 3 groups： the QRQYG group, the simvastatin group, and the control group. Sixteen weeks after administration of 0.9 g/kg QRQYG, 3 mg/kg simvastatin or 10 mg/kg sodium chloride per day to the respective groups, the animals were euthanized. The pathological morphologic changes in the vulnerable plaques were evaluated, the matrix metalloprotease-9 （MMP-9） expression was measured by immunohistofluorescence, the soluble intercellular adhesion molecule 1 （ICAM-1） was determined by ELISA, the nuclear factor kappaB （NF-κB） subunit p65 was measured by quantitative RT-PCR, and, finally, thrombospondin-1 （TSP-1） was determined by the immunohistochemical method. Results： The plaque cross-sectional area in the brachiocephalic artery （23.7%, P0.01）, the lipid core of the plaque （43.1%±3.1%）, and the number of buried ？brotic caps of the plaque were significantly decreased in the QRQYG group compared to the control group （both P0.01）; furthermore, the thickness of the ？brotic cap of the plaque increased and the intra-plaque hemorrhage of the plaque decreased. The serum soluble ICAM-1 （27.1±5.1 μg/mL）, the protein expression of MMP-9 and TSP-1 and the p65 mRNA expression increased in the QRQYG group in comparison with the control group （P0.05 or P0.01）. Conclusion： QRQYG could stabilize the vulnerable plaque through inhibition of the inflammatory response.     

Clinical Observation on the Treatment of Atrial Fibrillation with Amiodarone Combined with Shenmai Injection (参麦注射液)

Objective: To observe the therapeutic efficacy and safety of amiodarone combined with Shenmai Injection (参麦注射液) on atrial fibrillation. Methods: A total of 351 patients with atrial fibrillation caused by cardiovascular diseases and idiopathic atrial fibrillation were assigned to amiodarone group (control group, 128 cases) and amiodarone combined with Shenmai Injection group (treatment group, 223 cases). The patients in the control group received intravenous injection of 150 mg amiodarone in 10 min, followed by intravenous drip infusion at 1 mg /min and 6 h later at 0.5 mg /min until 48 h or cardioversion. The patients in the treatment group received the same treatment of amiodarone, while in addition, they received an injection of Shenmai Injection of 100 mL simultaneously. Blood pressure, ventricular rate, and cardioversion were observed. Results: The total efficiency rate was 98% (control group) and 99% (treatment group) (P0.05). The mean ventricular rate decreased 23% and 31% in the control group and the treatment group, respectively (P0.05). The mean cardioversion time of the two groups was 570±211 min and 351±123 min, respectively (P0.05). Only mild side effects were observed in both groups. Conclusion: Compared with amiodarone, amiodarone combined with Shenmai Injection takes effect more quickly with low side effects on the treatment of atrial fibrillation.     

Effects of Yinian Jiangya Decoction(颐年降压饮) Containing Serum on Cytokines Secretion of Vascular Endothelium of Spontaneously Hypertensive Rats

<正>Objective: To observe the effects of Yinian Jiangya Decoction (颐年降压饮, YNJYD) on cytokine secretion in spontaneoulsy hypertensive rats (SHRs) vascular endothelium. Methods: Aortic endothelial cells (ECs) were primarily cultured from SHRs; male SD rats were treated with different doses (high, medium, and low doses) of YNJYD, the blood was collected on the 21st day, and then, the serum was separated. ECs were cocultured with the serum for different time courses, and the culture supematant concentrations of endothelin (ET)-I, nitric oxide (NO), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor (PAl-l) were determined by ABC-ELISA methods. Results: ET-1, NO, t-PA, and PAl-1 levels in endothelial cell culture supernatant were increased in a time-dependent manner; YNJYD could significantly elevate NO and t-PA expressions in ECs, while ET-1 and PAl-1 expressions were dramatically decreased; these effects of YNJYD were in a concentration-dependent manner. Conclusion: The therapeutic effect of YNJYD on hypertension is attributed to its effect on regulating vessel dilation and blood coagulation, in which ET-1/NO and PAI-1/t-PA are two pairs of pivotal mediators.     

A randomized double-blind placebo-controlled study of Pu’er tea (普洱茶) extract on the regulation of metabolic syndrome

Objective:To explore the regulative efficacy of Pu’er tea(普洱茶)extract on metabolic syndrome. Methods:Ninety patients with metabolic syndrome were randomly divided into two groups,the intervention group administered with Pu’er tea extract,and the placebo group with placebo capsules.After 3 months’ treatment, body mass index,waist hip ratio,blood lipids,blood sugar,immune and inflammatory index,and oxidation index of the patients with metabolic syndrome were tested and analyzed.Results:In the intervention group,the body mass index,waist-hip ratio,fasting and 2 h postprandial blood glucose,serum total cholesterol,triglycerides, low density lipoprotein and apolipoprotein B-100 all decreased in the patients with metabolic syndrome,and also the high-density lipoprotein level increased and apolipoprotein A-1 showed the tendency to increase. Serum C-reactive protein,tumor necrosis factor-α,and interleukin-6 were decreased in the intervention group. lnterleukin-10 level was increased,MDA was decreased and superoxide dismutase was increased.Compared with before treatment and the placebo group,there were significant differences(P<0.05,P<0.01).Conclusions: Pu’er tea demonstrated excellent potential in improving central obesity,adjusting blood lipid,lowering blood sugar,regulating immunity and resisting oxidation.It can adjust the metabolic syndrome of different clinical phenotypes to different degrees,and is ideally fit for early prevention of metabolic syndrome.     

Protective effect of Renshen Yangrong Decoction (人参养荣汤) on bone marrow against radiation injury in mouse

Objective  

To explore the effect of Renshen Yangrong Decoction (人参养荣汤, RYD) in protecting bone marrow from radiation injury.