急性早幼粒白血病HL-60细胞动物模型的建立与鉴定

目的　建立人急性早幼粒白血病HL - 60细胞系重度联合免疫缺陷(SCID)小鼠移植瘤模型,探讨HL - 60细胞系在SCID小鼠体内的生物学行为。方法　分别在SCID小鼠腹腔或静脉注入5×1 0 6个白血病细胞,应用病理组织学、染色体核型分析等方法监测SCID小鼠外周血、肝、脾、骨髓中的白血病细胞。结果　腹腔或静脉注射均可在SCID小鼠体内形成白血病移植瘤。腹腔注射瘤块生长快,生存周期比静脉注射短。结论　SCID小鼠中白血病增殖和浸润程度反映了白血病细胞基本生物学特征,提示HL - 60 SCID小鼠模型为良好的肿瘤体内研究模型。     

移植性人白血病小鼠模型的建立

目的:建立移植性人白血病重度联合免疫缺陷(SCID)小鼠模型,探讨人白血病细胞在SCID小鼠体内的生物学行为,为研究药物等因素治疗白血病的体内方法奠定基础。方法:在SCID小鼠腹腔或静脉分别接种5×106～1×107个K562细胞,应用组织细胞化学、RT-PCR、流式细胞术、组织病理监测SCID小鼠的外周血、骨髓、肝脏、脾脏的白血病细胞。结果:K562细胞经静脉或腹腔途径均可植入SCID小鼠体内,形成白血病模型;3w时外周血即可见白血病细胞,肝、脾脏增生期细胞达40.11%、41.66%;发病晚期,模型动物骨髓细胞中CD13+细胞占10.15%;接种K562细胞后能检测到人DQα,ABL-BCR基因,接种K562细胞后3～4w即可见白血病细胞累及肝、脾、肾、骨髓等主要脏器。结论:SCID小鼠接种K562白血病细胞能成功建立白血病动物模型,CD13阳性率能快速、客观地判断模型中肿瘤细胞浸润程度。     

反义bcl-2基因抗白血病作用的研究

目的 研究反义bcl-2基因抗白血病作用及其特点,探讨反义基因或联合药物体外净化和治疗白血病可行性。方法 （1）RT－PCR和蛋白印迹方法检测HL－60、U937、K562、CEM白血病细胞株以及常见白血病类型的bcl-2 mRNA和P26蛋白表达情况。（2）选择高表达bcl-2基因的HL－60细胞株作为靶细胞,用人工合成bcl-2正反义硫代寡聚脱氧核苷酸（PS－ODN）体外培养处理。检测bcl-2 mRNA和P26蛋白的变化;DNA梯形分析细胞凋亡,锥虫蓝拒染法和克隆培养观察细胞生长;同时将HL－60细胞与PS－ODN培养孵育7天后的活细胞腹腔接种Scid鼠,半巢式RT－PCR和病理分析该HL－60细胞在体内生物学行为改变。（3）建立三种白血病细胞株CEM、K562、HL－60 Scid鼠白血病模型和有效监测与追踪白血病细胞株的方法,掌握体内白血病细胞株生物学行为;用bcl-2正反义PS－ODN腹腔给药治疗HL－60 Scid鼠白血病模型,观察治疗后体内白血病进展变化。（4）用RT－PCR方法克隆bcl-2基因和构建其两种不同的反义RNA逆转录病毒表达载体。（5）转染高表达bcl-2的HL－60细胞,比较两种反义RNA在对抗bcl-2的作用与降低内源性bcl-2蛋白水平上的异同性。观察两种反义RNA介导的靶基因抑制对烷化溶血磷脂（ALP）和四种化疗药物诱导的HL－60细胞凋亡的影响。（6）RT－PCR检测基因转染HL－60在化疗药物作用下FGFβ1表达。结果 （1）白血病细胞株表达bcl-2有差异,bcl-2 mRNA和它的P26蛋白不平行。（2）三种白血病细胞株HL－60,K562,CEM可在Scid鼠增殖,产生典型白血病浸润模型特征,白血病增殖和浸润程度与白血病细胞生物特性有关,其顺序为CEM＞K562＞HL－60。（3）人工合成的bcl-2反义硫化寡脱氧核苷酸（ASPO）能够下调HL－60细胞bcl-2表达,诱导凋亡,抑制增殖和克隆形成;同时使其失去在体内的致白血病能力。（4）bcl-2反义硫代寡脱氧核苷酸,腹腔给药能够抑制肿块增长,白血病扩张,延长Scid鼠生存期,呈剂量依赖性。（5）部分编码区的反义RNA能够降低HL－60细胞内源性Bcl-2蛋白水平,提高ALP和四种化疗药物对HL－60杀伤率,促进它们诱导的HL－60细胞凋亡。（6）反义基因转染的HL－60细胞在化疗药物作用下负调控因子TGFβ1基因表达增加。结论 bcl-2反义基因可有效对抗bcl-2原癌基因表达,发挥明显的抗白血病作用,为体外净化白血病细胞和体内治疗白血病提供依据。     

裸鼠高致瘤性人白血病细胞系染色体核型分析

目的 :探讨人白血病细胞系 K5 6 2 - n和 HL6 0 - n在裸鼠体内高致瘤性的细胞遗传学机制。 方法 :对裸鼠体内致瘤性不同的 K5 6 2细胞、K5 6 2 - n细胞及其衍生的各克隆株和 HL- 6 0细胞、HL6 0 - n细胞及其衍生的各克隆株 ,进行染色体核型分析。 结果 :K5 6 2细胞染色体数目变异范围为 44～ 71,干系核型为亚三倍体 ,染色体众数为 6 2± 2 ;裸鼠高致瘤性 K5 6 2 - n细胞系 ,干系为近二倍体细胞 ,染色体众数为 46 ;分析比较致瘤性不同的 K5 6 2、K5 6 2 - n细胞及其无致瘤性克隆株 K5 6 2 - n/ A的核型 ,发现 1、2、11、12、16、18、2 0、2 1、2 2等染色体的改变与 K5 6 2 - n细胞系的致瘤性增高有关。 HL- 6 0细胞染色体数目变异范围为 38～ 71,干系为近二倍体细胞 ,染色体众数为 45± 1;裸鼠高致瘤性 HL6 0 - n细胞系 ,干系核型转变为近三倍体 ,其染色体众数为 6 3± 1,HL6 0 - n细胞系衍生的 HL6 0 - A、HL6 0 - B、HL6 0 - E、HL6 0 - F各克隆株也都是近三倍体 ;分析比较 HL- 6 0细胞系及HL6 0 - n细胞系不同致瘤性克隆株的染色体核型变化 ,发现 Mar3、19、2 1、2 2、5号等染色体的变化与 HL6 0 - n细胞的高致瘤性相关。 结论 :K5 6 2 - n、HL6 0 - n细胞系致瘤性的增高涉及多条染色体的复杂变化。     

白血病MIC分型法鉴定SCID-人白血病模型

目的 运用白血病MIC分型法鉴定SCID-人白血病模型,探求SCID-人白血病模型建立的适宜条件.方法 ①按接种方式和鼠龄的不同将SCID小鼠平均分为4组.Ⅰ、Ⅱ组鼠龄小于5周,Ⅲ、Ⅳ组鼠龄大于5周.分别以静脉接种、腹腔接种(6～10)×106 HL-60细胞/鼠建立SCID-人白血病模型.不能发生白血病者用予60Co外照射后再接种.②运用白血病MIC分型法鉴定SCID-人白血病模型:形态学检查包括外周血、腹水涂片的细胞学检查和组织的病理学检查;免疫学检查采用流式细胞分析组织中CD33阳性率;分子遗传学检查骨髓细胞染色体.结果 ①鼠龄小于5周的Ⅰ、Ⅱ组的SCID小鼠接种HL-60细胞后4周,外周血白细胞上升至(10.±2.2)×109/L;2周时小鼠外周血中开始出现HL-60细胞,4周时比例达到(8.7±3.0)%,全部发生白血病.而鼠龄大于5周的Ⅲ、Ⅳ组小鼠小部分发病,未发病者予60Co 200 cGy/鼠,再接种2×106 HL-60细胞/鼠,也能全部发病.②腹腔接种者实体瘤的发生更为多见,大量腹水,恶病质明显;病理学检查和免疫学检查发现静脉接种者表现为脏器浸润较腹腔接种者广泛、明显,其全身弥漫性扩散的特点与人白血病的临床特点较为一致.③Ⅲ组小鼠经照射后再接种后发生的白血病在全身的广泛浸润程度较同样静脉接种的Ⅰ组更为明显.④鼠龄小于5周的静脉接种组(Ⅰ组)中一发病小鼠给予某不过血脑屏障的药物治疗后,发生了中枢白血病.结论 在适当控制SCID小鼠剩余的免疫防御系统后,采用静脉接种,数量为(6～10)×106/鼠,能发生全身弥漫性扩散性白血病,且还能模拟出化疗后庇护所白血病的发生.SCID-人白血病模型是一个体内研究人类白血病治疗良好的模型.     

Bcl-2参与SCID小鼠体内齐墩果酸诱导HL-60细胞凋亡的调控作用

目的:观察齐墩果酸(OA)对白细胞移植模型小鼠脾脏浸润白血病细胞凋亡数量和Bcl-2蛋白表达的影响,探讨OA对白血病模型鼠的治疗作用机制.方法:取浓度为2×107mL-1体外培养的人早幼粒系白血病HL-60细胞0.5 mL,腹腔注射重症联合免疫缺陷(SCID)小鼠,构建SCID小鼠的HL-60细胞移植瘤模型;模型成功后...     

人参多糖对移植性人白血病模型小鼠作用观察

目的建立重度联合免疫缺陷(SCID)小鼠人白血病模型,观察人参多糖在体内对白血病细胞的药理学作用。方法在SCID小鼠尾静脉分别接种K562细胞建立SCID小鼠人白血病模型,应用组织细胞化学、RT-PCR、流式细胞术、组织病理监测在给予人参多糖前后白血病模型SCID小鼠的外周血、骨髓、肝、脾中的白血病细胞。结果SCID小鼠人白血病模型外周血象在疾病早期无明显变化,第18天白细胞数略有升高,光镜下可检测到1%~4%人白血病细胞,病程后期,白细胞明显增高,血涂片检查人白血病细胞占10%;经GPS注射后,SCID小鼠一般状况明显好于对照组,其肺、肾等脏器的炎症及病理变化减轻。结论人参多糖在体内能有效地治疗髓系白血病,且未发现有明显的毒副作用。     

舟山眼镜蛇毒细胞毒素-Ⅻ的抗肿瘤作用

目的　了解舟山眼镜蛇毒细胞毒素 - (CTX- )的抗肿瘤作用。　方法　应用磺胺邻二甲氧嘧啶(SRB)法观察不同浓度 CTX- 对人肝癌 Bel74 0 4细胞和人早幼粒白血病 HL - 6 0细胞的生长抑制作用。观察CTX- 0 .4 ,0 .2 ,0 .1mg/ kg腹腔注射对 U 14荷瘤小鼠瘤块质量的影响并计算抑瘤率 ;CTX- 0 .4和 0 .2 m g/kg静脉注射对艾氏腹水癌 (EAC)小鼠腹水形成的影响并计算腹水减少率。　结果　 CTX- 可明显抑制人肝癌Bel74 0 4细胞和人早幼粒白血病 HL - 6 0细胞的生长 ,其 IC50 分别为 (1.12± 0 .13)和 (1.6 1± 0 .18)μg/ m L。 CTX- 0 .4 ,0 .2和 0 .1m g/ kg腹腔注射× 10 d,对宫颈癌 U14抑瘤率分别为 30 .5 0 % (P<0 .0 1) ,19.0 2 % (P<0 .0 5 )和12 .0 8% (P>0 .0 5 ) ;CTX- 0 .4和 0 .2 mg/ kg静脉注射× 3d对 EAC小鼠腹水形成减少率分别为 2 5 .5 % (P<0 .0 5 )和 14 .5 % (P>0 .0 5 )。　结论　舟山眼镜蛇毒细胞毒素 - 具有明显的抗肿瘤作用。     

急性淋巴细胞白血病CD34+CD38-细胞在NOD/SCID小鼠体内的自我更新与增殖能力

目的探索急性淋巴细胞白血病(ALL)患者CD34⁺CD38-细胞移植到NOD/SCID小鼠体内建立白血病的可行性、自我更新与增殖潜能。方法分选并鉴定ALL患者骨髓CD34⁺CD38-细胞及对照CD34-CD38⁺细胞后,经尾静脉分别注射10⁴个细胞于亚致死剂量射线照射的NOD/SCID小鼠体内,连续监测小鼠状态以及外周血血象改变,对濒死或死亡小鼠进行骨髓检查、肝脾病理学检查。结果接种从ALL患者分选的CD34⁺CD38-细胞到NOD/SCID小鼠体内后4周,小鼠外周血白细胞上升,到8周左右达高峰,约15×10⁹～20×10⁹/L,原始及幼稚淋巴细胞明显增多。骨髓象显示以原始及幼稚淋巴细胞增生为主,约为40%,且肝脾组织也有白细胞浸润,明显高于接种了对照组CD34-CD38⁺细胞的NOD/SCID小鼠。结论ALL患者CD34⁺CD38-细胞可以成功移植NOD/SCID小鼠,在小鼠体内增殖形成白血病,说明该群细胞具有自我更新和增殖的潜能,可作为探索白血病起始细胞研究的重要载体。     

尾静脉注射HL60、HL60/ADR细胞建立SCID beige小鼠白血病动物模型的研究

目的应用急性早幼粒细胞HL60及耐阿霉素(adriamycin,ADR)细胞HL60/ADR,构建SCID beige小鼠白血病模型。方法将15只4~5周龄的雌性SCID beige小鼠,随机分为对照组3只、四组模型12只(每组3只)。经2Gy X射线预处理,模型组分别经尾静脉接种对数生长期HL60细胞悬液1×10~7个/只(M1组)、5×10~6个/只(M2组),HL60/ADR细胞悬液1×10~7个/只(M3组)、5×10~6个/只(M4组),对照组鼠尾静脉注射等剂量生理盐水。观察一般情况,于照射前、造模第10、18、28天检测血常规、外周血白细胞分类、外周血CD33阳性细胞比例,第32天或濒死时处死取材,组织切片病理检查。结果各模型组于接种细胞第7天开始,出现竖毛、萎靡少动、脊背弓起、步态不稳、偏侧或转圈,M1、M3两组较M2、M4两组体重下降更明显(P0.05),显著低于对照组(P0.01)。至观察周期32 d,M1、M3两组生存率分别为33%、67%,M2、M4两组全部生存。各组小鼠经X线照射7 d内白细胞数迅速降低,随时间推移逐渐上升,第28天各模型组白细胞计数均显著高于对照组(P0.01);且各模型组白血病细胞比例亦明显升高,以接种HL60/ADR的M3组最为显著(P0.05);接种第28天时各模型组CD33阳性细胞比例显著高于对照组(P0.01),然各模型组间差异无显著性。第32天或濒死前取材,病理组织切片,造模各组脾均可见弥漫性白血病细胞浸润,肝偶见白血病细胞浸润灶。结论 SCID beige小鼠经2Gy X射线预处理后,每只尾静脉注射HL60、HL60/ADR细胞1×10~7个或5×10~6个均可构建急性髓系白血病动物模型,符合急性髓系白血病的生物学特点,高浓度成瘤更快,中位生存期约29 d。     

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.     

Development Is A High Level Inheritance for China Time-honored Brand Products

<正>As culture is the soul of a nation,the three books,"History of Chinese Philosophy,""History of Chinese Culture"and"History of Chinese Science and Technology"have demonstrated the superiority of our great Chinese culture,and its significant impact in the field of international culture.Both core value and practical value of the Chinese culture are very clear,reflected in the views of universe,nature,life,health,disease,and therapy and related considerations,concepts,and technical methods.