五倍子提取物抗鼻咽癌5-8F细胞的作用及机制

目的:探讨五倍子提取物鞣花酸抗鼻咽癌细胞的生物学活性及其分子机制。方法:体外培养鼻咽癌5-8F细胞,用2,4,6μg/mL鞣花酸处理48 h后,采用M实验分析细胞的增殖;Hoechst33258染色分析细胞凋亡;流式细胞仪分析细胞周期;Western印迹检测蛋白表达。结果:鞣花酸对5-8F细胞增殖有抑制作用,2,4,6μg/mL抑制率分别为(29.35±4.95)%,(53.32±4.44)%和(61.75±6.93)%,与对照组比较,差异有统计学意义(P<0.01);2,4,6μg/mL鞣花酸处理组S期细胞百分率分别为(25.47±0.74)%,(28.08±1.41)%和(35.49±0.66)%,与对照组(21.26±0.70)%比较,差异有统计学意义(P<0.01);随药物浓度增加,核固缩浓染的凋亡细胞明显增多,COX-2和stathmin蛋白表达下调。结论:五倍子提取物鞣花酸有抗鼻咽癌细胞的作用,其机制可能与COX-2和stathmin下调相关。     

冬凌草甲素抑制肝癌细胞HepG-2生长的实验研究

目的 探讨冬凌草甲素诱导肝癌细胞株HepG-2凋亡的作用及其机制.方法 MTT法检测细胞存活率;Annexin V和PI染色后流式细胞术检测HepG-2细胞凋亡率;Hoechst染色后荧光显微镜观察细胞形态;Western blotting检测MAPK信号通路凋亡相关蛋白的表达.结果 冬凌草甲素作用HepG-2细胞24 h后,可剂量依赖性减少细胞存活率,半数抑制率IC50为38.41 μmol/L.高、中、低浓度冬凌草甲素处理24 h后的流式结果显示,随浓度增加细胞凋亡率明显增加,HepG-2细胞可见典型的凋亡核改变.Western blotting结果证实冬凌草甲素可降低HepG-2细胞Bcl-2、caspase-9和caspase-3的蛋白表达,增加p-JNK、p-p38、p-p53和活化的caspase-9的蛋白表达.结论 冬凌草甲素可诱导肝癌HepG-2细胞凋亡,MAPK信号转导通路凋亡相关蛋白的活化与冬凌草甲素诱导HepG-2细胞凋亡相关.     

七叶皂苷对人肝癌HepG-2细胞增殖与凋亡的实验研究

目的：观察研究七叶皂苷对人肝癌HepG-2细胞的增殖周期及凋亡的影响,探讨七叶皂苷抗肿瘤作用的机制。方法：噻唑蓝（MTT）法检测七叶皂苷对人肝癌HepG-2细胞生长的抑制作用,流式细胞仪分析七叶皂苷对人肝癌HepG-2细胞周期及凋亡的影响。结果：七叶皂苷对人肝癌HepG-2细胞的增殖有明显的抑制作用,并随着作用浓度的增大,抑制作用逐渐增强。流式细胞仪检测可见亚二倍体峰,提示七叶皂苷有诱导人肝癌HepG-2细胞凋亡作用。七叶皂苷可通过诱导人肝癌HepG-2细胞凋亡并将细胞阻滞在G0~G1期,使细胞不能进入S期进行DNA合成,最终使瘤细胞的体外增殖受到抑制。结论：七叶皂苷抑制人肝癌HepG-2细胞的增殖,使细胞周期阻滞和诱导细胞凋亡,七叶皂苷可能在其抗肝癌作用中具有重要意义。     

人21.5kDa MBP基因对肝癌HepG-2细胞增殖与凋亡作用研究

目的 探讨人21.5kDa髓鞘碱性蛋白(MBP)对肝癌HepG-2细胞增殖与凋亡的影响.方法 用含21. 5kDa MBP基因的pSVCEP-MBP-CAT质粒转染肝癌细胞(HepG-2),设空质粒转染为对照组,用RT-PCR、Western blot杂交检测转染效果,后以H2O2处理上述细胞,通过MTT测定细胞增殖曲线,DNA琼脂糖电泳、免疫细胞化学分析、彗星电泳、TUNEL原位杂交等多种方法检测凋亡及相关蛋白的表达情况.结果 人21.5kDa MBP基因在肝癌HepG-2细胞转染成功, MTT测定细胞增殖曲线显示阳性转染组细胞有增殖和抗凋亡的作用,DNA琼脂糖电泳显示对照组有大量的DAN ladder形成,Caspase-3免疫组化证实对照组细胞Caspase-3表达显著升高,彗星电泳和TUNEL原位杂交显示对照组DNA损伤及凋亡严重而转染组较轻. 结论人21.5kDa MBP对肝癌HepG-2细胞有促进增殖和抗凋亡的作用.     

丹皮酚逆转内质网应激诱导的HepG2细胞凋亡抵抗

目的 探讨丹皮酚逆转内质网应激诱导的HepG2肝癌细胞凋亡抵抗的作用机制.方法 采用MTT及末端双脱氧核苷酸转移酶介导的dUTP缺口标记技术(TUNEL)观察内质网应激诱导剂衣霉素对正常人肝细胞株HL-7702及人肝癌细胞株HepG2细胞的增殖抑制及凋亡作用;Western blot法测定内质网应激诱导剂衣霉素对正常肝细胞HL-7702及肝癌细胞HepG2的环氧合酶-2(COX-2)表达的影响;采用Western blot法及TUNEL法观察COX-2的选择性抑制剂塞来昔布对内质网应激下的肝癌细胞HepG2 COX-2的表达及凋亡的影响;采用MTT及TUNEL法观察丹皮酚对内质网应激下的HepG2肝癌细胞的增殖抑制及凋亡作用;Western blot法观察丹皮酚对内质网应激诱导的HepG2肝癌细胞 COX-2表达的影响:结果与正常肝细胞HL-7702相比,在 TM诱导的内质网应激作用下肝癌细胞HepG2的增殖抑制率及凋亡率减少(P＜0. 05),并且内质网应激可诱导肝癌细胞HepG2产生COX-2蛋白.而丹皮酚可下调内质网应激诱导的HepG2肝癌细胞中COX-2的表达,并使内质网应激诱导的肝癌细胞的凋亡增加.结论 丹皮酚可通过下调COX-2的表达从而逆转内质网应激诱导的HepG2肝癌细胞凋亡抵抗.     

乌索酸通过抑制miR-21表达诱导肝癌HepG2细胞凋亡的机制

探讨乌索酸通过调控miR-21表达从而诱导肝癌HepG2细胞凋亡的作用机制。采用MTT方法检测乌索酸对肝癌细胞增殖的抑制作用;qPCR检测肝癌细胞中miR-21的表达水平及乌索酸对HepG2细胞中miR-21表达的调控作用;转染miR-21 mimics进HepG2细胞中上调miR-21的表达后,MTT、流式细胞检测法、RT-PCR方法分别分析miR-21在乌索酸对细胞的增殖、凋亡以及对凋亡相关基因的调控过程中的作用。结果显示,与肝正常细胞L-02以及肝癌SMCC-7721、Bel-7402细胞相比较,乌索酸对肝癌HepG2细胞的增殖抑制效果最强,且HepG2细胞中miR-21的表达水平最高。乌索酸可下调HepG2细胞中miR-21的表达,且在24 h的下调作用最强。miR-21的表达上调可以部分抵消乌索酸对HepG2细胞的抑制增殖及促进凋亡,部分抵消下调凋亡抑制基因Bcl-2、survivin表达和上调促凋亡基因Bax表达的作用。结果提示,乌索酸通过抑制miR-21的表达诱导肝癌HepG2细胞凋亡。     

乳浆大戟提取物对人胃癌和肺癌细胞增殖的影响

【目的】 探讨中药珍珠梅黄酮纳米粒(TTF1-NP)对人肝癌HepG-2细胞的内质网应激作用机制。 【方法】 通过MTT法观察不同时间(24、48、72h)、不同浓度(50、100、200 μmol/L)的TTF1-NP对人肝癌HepG-2细胞及正常肝细胞的增殖抑制影响;通过H-E、Hoechst和AO/EB染色观察TTF1-NP诱导人肝癌HepG-2细胞凋亡的形态学变化,利用流式细胞凋亡技术检测人肝癌HepG-2细胞的凋亡情况,蛋白质印迹和免疫细胞化学染色技术检测TTF1-NP诱导人肝癌HepG-2细胞凋亡的内质网应激起始蛋白GRP78及内质网应激凋亡蛋白CHOP、JNK及caspase-4的表达情况。 【结果】 MTT法检测细胞增殖抑制结果显示:不同浓度的TTF1-NP抑制人肝癌HepG-2的生长,呈一定的浓度和时间依赖关系。形态学检测结果:TTF1-NP作用后细胞数减少,形态改变,细胞变圆,变小,细胞核固缩,碎裂,细胞深染(H-E染色);随着TTF1-NP浓度增加,细胞染为红色(AO/EB染色);随着TTF1-NP浓度增加,细胞减少,细胞核内可见颗粒块状蓝色荧光(Hoechst染色)。流式凋亡检测结果:对照组细胞凋亡率为4.9%;50、100、200 μmol/L的 TTF1-NP作用人肝癌HepG-2细胞后,凋亡率为凋亡率为20.1%、86.2%、93.8%。蛋白质印迹检测结果显示,对照组细胞内CHOP、GRP78、JNK及caspase-4表达量很低,随着TTF1-NP浓度增加,上述蛋白表达逐渐升高,免疫细胞化学染色检测结果与蛋白质印迹检测结果基本相同。 【结论】 TTF1-NP可以诱导人肝癌HepG-2细胞,内质网应激途径是其诱导人肝癌HepG-2细胞凋亡的作用机制之一。     

COX-2 siRNA诱导人肝癌Hep3B细胞凋亡实验研究

目的:观察COX-2在人肝癌细胞Hep3B中的表达及小干扰RNA(small interfering RNA,siRNA)沉默COX-2基因对COX-2 mRNA和蛋白表达、细胞增殖及凋亡的影响。方法:将siRNA-COX-2质粒载体转染Hep3B细胞,WST-8法检测细胞增殖抑制情况,透射电镜观察凋亡小体出现情况;筛选单克隆细胞株Hep3B-sh,采用RT-PCR和Western blotting检测COX-2 mRNA和蛋白情况。结果:siRNA-COX-2能抑制Hep3B细胞的增殖,稳定转染pshRNA-COX-2可使Hep3B细胞中COX-2 mRNA及蛋白的表达明显减弱(P<0.01);透射电镜下可见凋亡小体出现,发生凋亡细胞数增加。结论:COX-2siRNA能有效抑制肝癌Hep3B细胞中COX-2的表达和细胞的增殖,促进凋亡发生,为肝细胞癌基因治疗提供实验依据和治疗靶点。     

雷公藤内酯醇对肝癌H22细胞增殖凋亡及COX-2表达的影响

目的 观察雷公藤内酯醇(TPL)对肝癌H22细胞增殖、凋亡及环氧合酶-2(COX-2)表达水平的影响. 方法 以体外培养的鼠源性H22细胞株为研究对象,实验分对照组和不同浓度的TPL处理组.MTT比色法测定TPL对H22细胞株增殖抑制情况;TPL处理48 h后,Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡情况,流式细胞术间接免疫荧光双标记法检测COX-2表达变化情况. 结果 随着药物浓度的增加及时间的延长,TPL能明显抑制H22细胞增殖.TPL诱导细胞凋亡及下调COX-2的表达具有浓度依赖性,不同浓度组比较,差别具有统计学意义(P<0.05). 结论 TPL可能通过诱导肝癌H22细胞凋亡及抑制COX-2表达发挥抗肿瘤作用.     

NS-398诱导肝癌细胞HepG2凋亡的作用机制

目的 探讨NS-398诱导肝癌细胞HepG2凋亡的分子机制.方法 采用MTT法测定COX-2选择性抑制剂NS-398对HepG2细胞的增殖抑制率;流式细胞仪检测凋亡;原位细胞凋亡检测法观察细胞凋亡形态学变化;免疫细胞化学分析COX-2、cIAP-1、XIAP、Survivin蛋白变化.结果 NS-398对HepG2细胞株有增殖抑制作用,流式细胞仪检测用药组凋亡率明显高于对照组.TUNEL染色可见典型的凋亡形态学特征.免疫细胞化学分析表明使用NS-398后,COX-2、cIAP-1、XIAP、Survivin蛋白表达降低.结论 COX-2选择性抑制剂NS-398对人肝癌细胞株HepG2有诱导凋亡作用,其机制可能通过抑制COX-2,下调cIAP-1、 XIAP、 Survivin的表达而诱导凋亡.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.