8-Br-cAMP和槲皮素对Eca-109细胞周期及凋亡的影响

目的观察8-Br-cAMP和槲皮素(quercetin)对Eca-109细胞周期及凋亡的影响.方法将贴壁培养的Eca-109细胞随机分成3组,Br组加8-Br-cAMP至终浓度为2×105 mol/L;Q组加槲皮素至终浓度为43 μmol/L;C组不加任何药物,只加入新培养液培养.3组在同等条件下继续培养48 h.采用流式细胞仪检测细胞的周期,采用TUNEL法检测细胞凋亡百分率,采用DNA琼脂糖凝胶电泳法观察细胞DNA降解情况.结果2实验组细胞中G2-M期细胞比例较对照组显著上升(P<0.01).细胞凋亡百分率2实验组显著高于对照组(P<0.01);Br组和Q组相比,差异无统计学意义(P>0.05).DNA琼脂糖凝胶电泳中Br组和Q组皆呈DNA"梯样型"带,对照组未呈现DNA降解.结论8-Br-cAMP和槲皮素皆可阻滞Eca-109细胞于G2-M期,并进一步诱导Eca-109细胞凋亡.     

8-Br-cAMP和槲皮素对Eca-109细胞凋亡中P38和Caspase-3表达的影响

目的 探讨8-Br-cAMP和槲皮素对Eca-109细胞凋亡中P38、Caspase-3表达的影响.方法将贴壁培养的Eca-109细胞随机分为3组①对照组只加DMEM(Sigma)培养液(含体积分数10%胎牛血清)培养48 h.②Br组终浓度为2×10-5 mol/L 8-Br-cAMP的培养液培养48 h.③Q组终浓度为43 μmol/L槲皮素的培养液培养48 h.对以上3组细胞以TUNEL法染色计数细胞凋亡率.以免疫组化方法观察3组细胞的P38 MAPK-IR和Caspase-3-IR反应性.结果Br组及Q组细胞凋亡率高于对照组;Br组及Q组细胞的P38 MAPK-IR高于对照组;Br组及Q组细胞的Caspase-3-IR亦高于对照组.P38 MAPK-IR与Caspase-3-IR呈正相关.P均<0.05.结论P38 MAPK及Caspase-3参与了8-Br-cAMP及槲皮素诱导的Eca-109细胞的凋亡.     

8-Br-cAMP和槲皮素对Eca-109细胞核基质蛋白的影响

目的探讨8-Br-cAMP和槲皮素对Eca-109细胞核基质蛋白的影响.方法将贴壁培养的Eca-109细胞随机分成3组,Br组加8-Br-cAMP至终浓度为2×105 mol/L;Q组加槲皮素至终浓度为43 μmol/L;C组不加任何药物,只加入新培养液培养.同时培养48 h,应用SDS-PAGE技术分析3组细胞核基质蛋白成分.结果实验组(Br组与Q组)与对照组(C组)比较,相对分子质量64 000、66 000、58 000为增加的条带,14 000、17 000、18 000为增强的条带,92 000是减弱的条带.2个实验组相比,以Br组变动显著.结论8-Br-cAMP和槲皮素可以诱导Eca-109细胞核基质蛋白成分改变,可能对肿瘤细胞基因表达调控、分化及凋亡起重要作用.     

8-Br-cAMP和槲皮素对Eca-109细胞相关基因的DNA拷贝和基因表达的影响

目的探讨8-Br-cAMP和槲皮素对Eca-109细胞wtp53、c-myc、bcl-2基因的扩增、表达以及VEGF表达的影响.方法将贴壁培养的Eca-109细胞随机分成3组,Br组加8-Br-cAMP至终浓度为2×105 mol/L;Q组加槲皮素至终浓度为43 μmol/L;C组不加任何药物,只加入新培养液培养.3组细胞同时培养48h.以cDNA 阵列技术显示Eca-109细胞凋亡中wtp53、c-myc、bcl-2 DNA拷贝和mRNA的表达,以免疫组化法显示VEGF的表达.结果Br组和Q组与C组相比wtp53 DNA拷贝和mRNA信号较强,c-myc、bcl-2 DNA拷贝和mRNA信号较弱,VEGF表达较弱.结论8-Br-cAMP和槲皮素皆可通过上调wtp53,下调bcl-2、c-myc基因的扩增和表达,下调VEGF的表达,而诱导Eca-109细胞凋亡.     

8-Br-cAMP和槲皮素单独和联合应用对Eca-109细胞POLB及多药耐受相关蛋白的影响

目的探讨8-Br-cAMP和槲皮素单独和联合应用对Eca-109细胞耐药性的影响.方法将培养的Eca-109细胞随机分为4组①8-Br-cAMP组(Br组); ②槲皮素组(Q组); ③(Br+Q)组; ④不加药物的对照组(C)组.同时培养48 h,对以上4组细胞制备细胞滴片及硝酸纤维素膜(NCM)滴膜2种标本,分别进行MRP和POLB的免疫组化染色和免疫斑点印迹阵列及TLC扫描.结果免疫组化染色显示 C组的MRP-IR、POLB-IR强于Br组、Q组或(Br+Q)组,P<0.01.TLC扫描OD值C组MRP-IR高于Br、Q(Br+Q)组约3倍;POLB-IR高于Br、Q或(Br+Q)组约2倍;MRP与POLB呈显著正相关r=0.983 8,P<0.01.结论8-Br-cAMP和槲皮素联合用药或单独用药均可降低Eca-109细胞的多药耐受性而提高药物的敏感性,可有利于提高临床疗效.     

8-Br-cAMP和槲皮素对Eca-109细胞p16基因启动子DNA结合蛋白及其mRNA表达的影响

目的应用Southwestern技术研究8-Br-cAMP和槲皮素(quercetin)对Eca-109细胞DNA结合蛋白与p16基因启动子区的结合调控mRNA表达的影响.方法将贴壁培养的Eca-109细胞随机分成3组,Br组加8-Br-cAMP至终浓度为2×105 mol/L;Q组加槲皮素至终浓度为43 μmol/L;C组不加任何药物,只加入新培养液培养.同时培养48 h,应用质粒提取及限制性酶切回收目的DNA片段,采用生物素随机引物标记探针;应用DNA斑点印迹技术检测探针的灵敏度后以Southwestern印迹技术检测与p16基因启动子区特异性结合的DNA 结合蛋白(DBP);以原位杂交检测p16基因mRNA的表达情况.结果实验组条带强于对照组,Br组强于Q组;核基质DNA结合蛋白主带位于Mr66 000、58 000和20 000;核外DNA结合蛋白主带位于Mr40 000、28 000和20 000;2者中20 000是共同成分.原位杂交技术显示p16基因mRNA蓝紫色信号颗粒位于胞质,杂交信号以实验组较强.结论8-Br-cAMP和quercetin尤其是前者作用于人食管癌Eca-109细胞,诱导产生与p16基因启动子结合较强的DBP,启动p16基因表达.     

8-Br-cAMP对Eca-109细胞c-myc，wtp53，iNOS基因和EGFR表达的影响

目的探讨8-Br-cAMP对Eca-109细胞c-myc、wtp53、iNOS基因和EGFR表达的影响.方法将贴壁培养的Eca-109细胞随机分为2组①实验组加8-Br-cAMP至终浓度为2×10-5mol/L,培养24 h;②对照组不加8-Br-cAMP,培养24 h.对2组的细胞涂片和硝酸纤维素膜(NCM)标本进行c-myc、wtp53、iNOS基因和EGFR表达检测,并对扫描数值进行统计学处理.结果原位杂交及免疫组化反应信号皆定位于Eca-109细胞的胞质,扫描数值显示①c-myc mRNA实验组为3.38±0.99,对照组为5.18±1.39;②wtp53 mRNA实验组为2.74±0.83,对照组为0.38±0.27;③iNOSmRNA实验组为4.52±0.74,对照组为2.63±0.13;④EGFR-IR实验组为2.37±1.05,对照组为4.38±0.48.以上实验组与对照组比较差异有统计学意义(P均<0.05).结论c-myc、wtp53和NO均可参与8-Br-cAMP诱导癌细胞的分化效应.     

8-Br-cAMP和槲皮素单独和联合应用对Eca-109细胞凋亡和Bcl-2、Bax、XRCC1等凋亡相关蛋白的作用

目的探讨8-Br-cAMP和槲皮素联合应用对Eca-109细胞的凋亡和Bcl-2、Bax、XRCC1等凋亡相关蛋白的作用.方法将Eca-109细胞随机分为4组①8-Br-cAMP组(Br组); ②槲皮素组(Q组); ③(Br+Q)组; ④不加药物的对照(C)组.同时培养48 h,对以上4组细胞制备细胞滴片及硝酸纤维素膜(NCM)滴膜2种标本,分别进行Bcl-2、Bax、XRCC1的免疫组化和免疫斑点印迹实验,并以TUNEL法检测各组细胞凋亡率.结果细胞凋亡率,C组为4%,(Br+Q)组为70%,Br组为42%,Q组为35%,(Br+Q)组>Br组或Q组>C组,P<0.001.Br、Q及(Br+Q)组与对照组相比,Bcl-2-IR及XRCC1-IR皆低于C组,P<0.001,而Bax-IR则高于C组,P<0.001;Br组Q组或Br组(Br+Q)组,P>0.05.Bcl-2-IR与XRCC1-IR呈显著正相关,与Bax-IR和Bax-IR与XRCC1皆呈显著负相关.结论Br与Q联合用药与单独用药皆可下调Bcl-2及XRCC1表达,上调Bax表达;联合用药的细胞凋亡率显著高于单用Br或Q.提示,中西医联合用药可能更具有临床治疗意义.     

8-Br-cAMP和槲皮素对Eca-109细胞DNA polβ基因及相关基因表达的影响

目的探讨8-Br-cAMP及槲皮素等分化诱导剂对Eca-109细胞DNA polβ基因及相关基因表达的影响.方法将培养的Eca-109细胞分为3组①加8-Br-cAMP组(Br组);②加槲皮素组(Q组);③不加任何药物对照组(C组).同时培养48 h,将野生型DNA polβ的cDNA, EGFR cDNA,野生型(wt) p53 cDNA及c-myc cDNA分别制备了生物素/地高辛标记的探针进行原位杂交和RNA斑点印迹阵列.另进行POLB,XRCC1,VEGF及PCNA的免疫组化染色及免疫斑点印迹阵列.结果Br组和Q组的DNA polβ,EGFR,c-myc基因表达下调而wtp53基因表达上调.POLB,XRCC1,PCNA及VEGF免疫斑点印迹减弱.8-Br-cAMP及槲皮素显著下调Eca-109细胞的DNA polβ基因表达及相关癌基因表达,同时上调抑癌基因的表达.作为POLB的异二聚体XRCC1及恶性细胞生物标志的PCNA及VEGF表达下调.结论分化诱导剂下调Eca-109细胞DNA polβ基因表达,提示Eca-109细胞的polβ基因是高表达的;功能调整后的polβ通过相关基因表达的改变尤其是wt p53基因表达的上调在癌细胞分化中可能起重要作用.     

8-Br-cAMP和槲皮素对Eca-109细胞iNOS基因拷贝及Caspase-3表达的影响

目的 :观察 8 Br cAMP和槲皮素对Eca 10 9细胞iNOS基因拷贝及Caspase 3表达的影响。方法 :将贴壁培养的Eca 10 9细胞随机分成 3组 ,Br组 :加 8 Br cAMP至终浓度为 2× 10 5mol/L ;Q组 :加槲皮素至终浓度为 4 3μmol/L ;C组 :不加任何药物 ,只加入新培养液培养。 3组细胞同时培养 4 8h。采用原位斑点印迹法显示iNOS基因拷贝和mRNA的表达 ,采用细胞免疫化学技术显示Caspase 3的表达。结果 :Br和Q组较C组iNOS基因拷贝和mRNA表达信号较强。Br和Q组细胞内Caspase 3免疫反应 (IR)性较强 ,Br和Q组与对照组相比 ,存在显著性差异 (P 0 .0 1) ;Br组的Caspase 3 IR强于槲皮素组 (P <0 .0 1)。结论 :8 Br cAMP和槲皮素皆可上调iNOS基因的扩增和表达 ,最终可都通过上调Caspase 3的表达 ,诱导Eca 10 9细胞凋亡。     

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…     

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.     

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.     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.