蜂毒素的抗肿瘤生物活性及其稳定性研究

目的：通过中压液相色谱提取、纯化蜂毒素，研究其抗肿瘤生物学活性，及其稳定性。方法：通过中压液相色谱提取蜂毒素，经HPLC检测纯度，采用MTT法，体外观察蜂毒素对4种肝癌细胞系增殖的抑制作用，同时与蜂毒素标准品进行抗肿瘤生物活性的比较，并比较不同批次提纯蜂毒素的抗肿瘤作用。结果：蜂毒素在16～128μg/ml对肝癌细胞增殖有抑制作用，并呈剂量依赖性，对人正常肝脏来源细胞株L-02细胞抑制作用不明显。在60μg/ml浓度，各批次提取的蜂毒素与蜂毒素标准品间抗肿瘤活性无显著性差异。结论：采用中压液相色谱提取、纯化的蜂毒素具有抗肿瘤生物活性，并且稳定性良好。     

蜂毒素对小鼠肝癌H22细胞增殖的影响

目的 观察蜂毒素对小鼠肝癌H22细胞增殖的影响，探讨其抗肿瘤的可能作用环节。方法 以MTT法测定蜂毒素体外对小鼠肝癌H22细胞增殖的影响，运用流式细胞术分析增殖细胞核抗原(PCNA)的表达，并观察iv蜂毒素对荷瘤小鼠的抑瘤作用。结果蜂毒素对H22细胞的抑制作用随药物浓度增加而增强；蜂毒素作用于H22细胞后，细胞PCNA平均荧光强度低于对照组；iv蜂毒素对小鼠皮下H22肝癌细胞具有明显的抑制作用，抑瘤作用随浓度增加而增强。结论蜂毒素在体内外对小鼠肝癌H22细胞增殖均有明显的抑制作用，下调细胞PCNA表达可能是其作用环节之一。     

蜂毒素缓释制剂瘤内注射减毒增效作用的实验研究

目的：观察蜂毒素缓释制剂瘤内注射的减毒增效作用。方法：以低、中、高3种剂量（0．16、0．32和1．6mg/kg）的蜂毒素－泊洛沙姆407缓释剂对H22荷瘤小鼠施瘤内注射后,以小鼠生存期、血细胞、肝功能及其肿瘤病理形态变化为指标观察其体内抗肿瘤作用。结果：（1）蜂毒素缓释制剂的毒性明显低于单纯蜂毒素（P＜0．05）;（2）蜂毒素缓释剂型治疗组肿瘤生长受到显抑制,低、中、高剂量的抑瘤率分别为20．7％、36．5％、53．3％;肿瘤组织广泛坏死,治疗后平均生存期明显延长（P＜0．05）,综合疗效优于单纯蜂毒素组。结论：蜂毒素缓释制剂瘤内局部注射能够通过延缓蜂毒素的释放速度,增加其与肿瘤细胞直接作用的时间等途径达到减毒增效的目的。     

卟啉氮芥对人肝癌细胞SMMC7721的光动力杀伤作用

目的：研究新单体化合物卟啉氮芥的光化疗抗肿瘤作用。方法：用结晶紫色法测定 啉氮芥对人肝癌细胞SMMC7721的光动力抑制效应。用测定培养上清乳酸脱氢酶（LDH）活性方法研究光动力作用对人肝癌细胞SMMC7721细胞膜的作用,用^3H-Td掺入法研究光动力作用对人肝癌细胞SMMC7721DNA合成的影响。结果：卟啉氮芥（5μg/ml,培养1h）可使人肝癌细胞SMMC7721D值明显降低、经光激发可使培养上清LDH活性显升高;可使体外培养从肝癌细胞SMMC7721的^3H-TdR掺入显下降,经光激发可使体外培养人肝癌细胞SMMC7721的^3H-TdR掺入量进一步显下降。结论：卟啉氮芥可显抑制人肝癌细胞SMMC7721DNA的合成;对人肝癌细胞SMMC7721有显的光动力抑制作用,轲破坏肿瘤细胞的细胞膜,抑制肿瘤细胞DNA的合成,提示卟啉氮芥对肿瘤细胞有显的光动力杀伤作用,具有发展为光化疗抗癌新药的前景。     

绿茶儿茶素与抗癌药联合应用对二株人肝癌细胞生长的影响

目的：研究绿茶儿茶素（GTC）与抗癌药阿霉素（ADM）,5－氟尿嘧啶（5－Fu)和阿糖胞苷（AraC)联合应用对人肝癌细胞体外生长的抑制作用。方法：应用MTT法检测GTC先于抗癌药,抗癌药作用后或与抗癌药同时作用,对BEL－7402和SMMC－7721人肝癌细胞的生长抑制作用。结果：GTC与抗癌药有相加性抑瘤效应。结论：GTC很可能具有临床应用前景。     

胡桃楸树皮提取物对SMMC-7721、MCF-7和A549肿瘤细胞的抑制作用及其机制

目的：观察胡桃楸树皮提取物（JMME）对SMMC-7721、MCF-7和A549 3种肿瘤细胞的生长抑制作用,确定其抑瘤的药用价值,通过检测JMME诱导肿瘤细胞凋亡、抑制端粒酶活性变化阐述其抑瘤机制。方法：分别以25、50、100、200、400和800 mg•L^-1JMME作用于体外培养的SMMC-7721、MCF-7和A549肿瘤细胞72 h,采用MTT法检测生长抑制率;以200 mg•L^-1JMME作用于体外培养的SMMC-7721、MCF-7和A549,于倒置显微镜下观察细胞形态学变化;琼脂糖凝胶电泳法检测细胞凋亡;PCR-ELISA法检测端粒酶活性。结果：不同浓度JMME（25、50、100、200、400和800 mg•L^-1）对SMMC-7721的细胞生长抑制率(%)分别为13.06、21.77、29.88、41.74、69.82和78.08;对MCF-7的抑制率(%)为24.91、38.63、49.72、61.84、65.69和81.28;对A549的抑制率(%)为12.32、26.21、41.65、55.38、62.87和80.13;与对 照组比较,JMME各浓度作用的3种肿瘤细胞的生长抑制率均升高（P<0.05）,抑制率随JMME对SMMC、MCF-7和A549细胞的浓度的增加而增高（P<0.05）。JMME对SMMC、MCF-7和A549细胞的半数抑制浓度（IC₅₀）分别为211.21、111.07和176.20 mg•L^-1,对MCF-7的IC₅₀小于其他两种瘤细胞株。经不同浓度JMME作用单位视野内细胞数量明显减少,细胞间隙增大,细胞体积明显缩小,细胞失去原有的形态,胞体皱缩变圆,出现膜发泡现象,对照组无此变化。提取DNA进行琼脂糖凝胶电泳,见DNA Ladder,表明3种肿瘤细胞经不同浓度JMME作用后出现不同程度凋亡。3种肿瘤细胞与阴性对照组比较端粒酶活性升高,而经过JMME作用后端粒酶活性均受到了抑制,抑制率分别为42.86%、73.30%和58.78%。结论：胡桃楸树皮提取物在体外有抗肿瘤活性,其机制可能与其诱导肿瘤细胞凋亡﹑抑制端粒酶活性有关。     

人DNA MTase反义基因转染对诱导肝癌SMMC-7721细胞凋亡的影响

目的；观察人DNA MTase反义基因转染对肝癌细胞系SMMC－7721凋亡的影响。方法：采用脂质体法将构建的包含正，反义DNA MTase基因片段的真核表达载体转染入肝癌细胞系SMMC－7721；采用MTT法绘制生长曲线；流式细胞法和原位末端标记法检测细胞凋亡改变。结果：PCR法扩增外源性NeoR基因证实用脂质体法成功将重组载体转染入细胞；生长曲线显示转染反义DNA MTase基因片段的SMMC－7721细胞增殖速度减慢；流式细胞法测得其凋亡率由3．1％增加到13．5％，末端标记法也显示其调亡指数增加。结论：人DNA MTase反义基因转染可诱导肝癌细胞系SMMC－7721凋亡。凋亡基因功能被恢复可能主要原因之一。     

金边瑞香对SMMC7721,NKM45,A549,S180肿瘤细胞增殖的影响

[摘要] 目的 探讨金边瑞香对SMMC7721, NKM45, A549, S180肿瘤细胞增殖的影响。方法 采用流式细胞术，观察不同浓度金边瑞香浸膏对SMMC7721细胞增殖周期的影响；采用MTT法检测不同浓度金边瑞香浸膏对SMMC7721, NKM45, A549, S180肿瘤细胞株增殖作用的影响。结果 ① 金边瑞香浸膏可使SMMC-7721细胞增殖周期G0～G1细胞增多，S期细胞减少，且0.5g/ml金边瑞香浸膏作用最明显，G0～G1期细胞达80.64%；1g/ml，G0～G1期达到72.19%。②金边瑞香浸膏浓度在0.03125~1g/ml范围内对体外培养的SMMC7721, NKM45, A549, S180肿瘤细胞生长均有一定程度的抑制作用，其抑制作用与剂量成正相关，0.5g/ml金边瑞香浸膏对SMMC7721, NKM45, A549, S180肿瘤细胞生长抑制率为65.82%, 60.38%, 42.36%, 49.94%，但随着浓度增加,这种抑制作用反而减弱，在1g/ml其生长抑制率均有所减弱。结论 金边瑞香能使体外培养的SMCC7721细胞周期阻滞在G0～G1期，金边瑞香对体外培养的SMMC7721, NKM45, A549, S180肿瘤细胞株的增殖均有较强的抑制作用。     

Nrf3基因对肝癌SMMC7721细胞系的体外研究

目的在体外检测Nrf3基因对肝癌SMMC7721的生长影响作用。方法构建其融合蛋白真核表达载体,脂质体介导该真核表达载体转染肝癌SMMC7721细胞系,FCM观察其在体外对肝癌SMMC7721细胞系细胞周期和凋亡的影响;荧光显微镜观察候选基因亚细胞定位。结果Nrf3在肝癌SMMC7721细胞中表达,荧光显微镜下观察Nrf3定位于细胞核内。FCM分析显示Nrf3影响下肝癌SMMC7721G2/M＋S期细胞所占比例较对照组明显减少,G0/G1细胞所占比例明显增加。证实Nrf3可抑制肝癌SMMC7721的DNA合成和有丝分裂,促使细胞阻滞于G0/G1期,抑制肝癌SMMC7721细胞的体外生长。FCM分析,未观察到明显的＂亚G1＂峰（即凋亡峰）,提示Nrf3在体外对肝癌SMMC7721细胞的凋亡无影响。结论Nrf3在体外具有抑制肝癌细胞增殖的功能,对肝癌细胞的凋亡无影响,为肿瘤抑制基因。     

蜂毒素对人肝癌细胞生长的抑制作用及其机制探讨

目的：研究蜂毒素对人肝癌细胞( HepG2、SMMC-7721)生长的抑制作用及其机制。方法采用MTT法检测蜂毒素对HepG2和SMMC-7721细胞增殖的抑制作用,实时定量PCR检测细胞中microRNA-203(mir-203)以及PIK3CA mRNA 的表达变化, Western blot法检测磷脂酰肌醇3-激酶(PI3K)、磷酸化蛋白激酶B(P-AKT)的表达变化,瞬时转染人mir-203 mimics 过表达 mir-203后应用实时定量 PCR 和Western blot 检测 mir-203对 PI3K/AKT 信号通路的作用。结果与正常组比较,蜂毒素处理组细胞增殖明显受到抑制,并呈浓度依赖性;实时定量PCR结果显示蜂毒素(1、2、4 mg/L)可以明显升高mir-203的表达,PIK3CA mRNA 的表达无明显改变;Western blot法结果显示蜂毒素(1、2、4 mg/L)可以降低 PI3K 以及 P-AKT 蛋白的表达。在 HepG2和SMMC-7721细胞中过表达mir-203后,与正常组及转染阴性对照组比较,PIK3CA mRNA的表达无明显改变但PI3K以及P-AKT的蛋白表达降低。结论蜂毒素能抑制 HepG2和SMMC-7721细胞的增殖,其机制可能是通过上调mir-203的表达,进而在转录后水平靶向抑制PI3K的表达,抑制PI3K/AKT信号通路的活化。     

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.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.