自体树突状细胞用于慢性粒细胞白血病骨髓净化的初步研究

目的 考察慢性粒细胞白血病（CML）树突状细胞（DC）刺激自体骨髓T细胞,清除白血病细胞的作用。方法 采用3例CML病人（2例慢性期和1例急变期）外周血,分离单个核细胞（PBMNC）,经塑料贴壁后在含自体血浆、重组人白介素4（rhIL－4)、重组人粒－单集落刺激因子（rhGM－CSF）和重组人肿瘤坏死因子（rhTNFα）的RPMI－1640培养液。37℃CO2孵箱培养10天,或用免疫磁珠从PBMNC分离DC,在含自体血浆、rhGMCSF和rhTNFα的RPMI1640培养液培养3天,收获的细胞即DC加到含rhIL－2的Dexter骨髓长期培养体系,置37℃,5％CO2孵箱培养7天,观察前后骨髓细胞的免疫表型和P210阳性细胞比例。结果 从CML病人外周血获得的DC大多数表达P210。在含rhIL－2的Dexter体系加入DC能进一步增加体系中的T细胞数。在2例慢性期病人体系中的P210。在含rhIL－2的Dexter体系加入DC能进一步增加体系中的T细胞数。在2例慢性期病人体系中的P210阳性细胞减少,但在急性期病例,体系中P210细胞明显增加。结论 CML病人的DC表达P210,能刺激自体骨髓中T细胞增生,在慢性期病人,这些自体DC活化的骨髓T细胞有清除白血病细胞的作用。     

类风湿关节炎外周血和关节液树突状细胞的膜分子表达及功能研究

目的研究类风湿关节炎（rheumatoid arthritis,RA）患者树突状细胞（dendritic cell,DC）的膜分子表达水平和功能的变化,探讨DC在RA发病机制中的作用。方法应用流式细胞术、混合淋巴细胞培养测定诱导DC细胞表面分子表达、对同种异体T细胞功能情况,诱导成熟后DC表面分子表达与临床活动及预后相关指标ESR、CRP、RF、Stoke指数的相关性。结果RA外周血来源DC表达CD83、CD209、CD80、HLA—DR较正常人外周血来源DC增高（P〈0．05）,RA关节液来源DC较RA外周血来源DC表达CD83、CD209、CD86、CD80明显增高（P〈0．05）。RA关节液组较RA和正常人外周血DC对同种异体T细胞的刺激明显增强。RA患者外周血诱导成熟DC表型CD83、CD209与stoke评分指数呈正相关。结论提示RA时关节内免疫反应可能与局部DC分化、成熟能力增强有关,后者可通过激活T细胞引起自身免疫反应,造成滑膜组织损伤。RA患者DC表面共刺激分子表达水平的增高可能与患者免疫亢进及所致的免疫损伤有关。     

树突状细胞体外培养体系的优化

目的：探讨无血清专用培养基从人外周血单个核细胞（DC）分离培养树突状细胞的优化方法。方法：取正常成人新鲜血液，经密度梯度离心法，利用连续贴壁的方法获得单个核细胞，采用无血清培养基经人重组粒细胞-巨噬细胞集落刺激因子（rhGM-CSF）、重组人白介素-4（rhIL-4）和肿瘤坏死因子-α（TNF—α）诱导产生成熟DC，倒置显微镜观察树突状细胞结构，流式细胞仪检测DC表面标志物表达水平。结果：连续贴壁法无血清专用培养基体外分离培养人外周血DC，所获得DC数量和纯度均都多于以往一次贴壁分离含10％胎牛血清RPMI培养基培养的DC。培养1周的DC高表达CD83、HLA-DR、CD80、CD86、CD83^＋HLA—DR^＋及CD80^＋CD86^＋。结论：采用无血清专用培养基连续贴壁法可分离培养数量较大、纯度较高的人外周血来源的DC。     

慢性粒细胞白血病树突状细胞的体外定向诱导扩增及鉴定

目的：W本外定向诱导扩增慢性粒细胞白血病（CML）患者外周血单个核细胞（PBMNC）来源的树突状细胞（CML－DC）并测定其表型。方法：取CML患者新鲜抗凝血,用淋巴细胞分离液分离PBMNC,经细胞因子rhGM-CSF,rhIL-4,rhTNF-α联合刺激培养9－12d,收集培养细胞,光镜形态学分析并细胞计数,结合PE－CD1a单抗、FITC－CD14居流式细胞 （FCM）上进行CML－DC比例及表型分析。结果CML患者PBMNC经以上细胞因子联合培养,CML－DC含是可达3．5％－23．5％,而新鲜分离的CML患者PBMNC中CML－DC含量仅为0．1％－1％,结论CML患PBMNC 体外定向诱导扩增培养可以生成较多的CML－DC,从而为CML的免疫治疗应用于临床提供了基础。     

乳腺癌患者外周血中树突状细胞表面标记CD1a、CD83的研究

目的：对乳腺癌患者外周血中培养的树突状细胞（dendritic cells,DC)表面分子CD1a,CD83的表达进行研究。方法：采用密度梯度离心的方法分离人外周血中的单个核细胞，用粒细胞-单核细胞集落刺激因子（GM-CSF），白细胞介素-4（IL-4），肿瘤坏死因子α（TNF-α）细胞因子组合，刺激DC增殖，分化，用标有荧光素的单抗标记培养细胞，在流式细胞仪上分析所培养的细胞。结果：该细胞表达CD1a,CD83分子，但CD1a,CD83在CD3^ T,CD19^ B淋巴细胞上也表达，CD1a,CD83在活化的淋巴细胞上表达水平比未活化的高，CD19^ B细胞上表达水平比CD3^ T细胞高。结论：CD1a,CD83等分子并非DC特有的标记，目前鉴定DC仍然要靠细胞形态，细胞表达CD1a,CD83以及共刺激分子等综合因素来确定DC。     

人外周血中树突状细胞的诱导、培养及其表面标记的初步研究

目的：在人外周血中有效地诱导，培养树突状细胞（dendritic cells,DC)并鉴定其表面分子的表达，方法：采用密度梯度离心的方法分离人外周血中的单个核细胞，在6孔板上贴壁2h，然后吸去悬浮细胞，用粒细胞－巨噬细胞集落群体刺激因子（GM－CSF）、白细胞介素－4（IL－4）、肿瘤坏死因子－α（TNF-α）细胞因子组合，刺激树突状细胞（DC）增殖，分化。用标有荧光素的单抗标记培养细胞，在激光共聚共焦显微镜下拍摄标记有荧光素的细胞，在流式细胞仪上分析所培养的细胞。结果：该细胞直径在10－20μm，培养14天后，CD1a^ 48%,CD83^ 97%,CD40^ 33%，人体主要组织相容性复合物（MHC）Ⅱ^ 95%,CD80^ 98%,CD86^ 93%,CD83是DC的成熟标记。结论：该方法是一种可靠的诱导和培养DC的方法，对于乳腺癌患者还可通过动员等措施进一步提高DC的分离率。     

人脐血CD34+细胞和贴壁细胞体外诱导树突状细胞及其鉴定

目的 从人脐血中不同的前体细胞扩增树突状细胞(DCs)，并对其进行鉴定。方法 用免疫磁珠法分离人脐血CD34^ 细胞，以GM—CSF、TNF-α、FL、SCF和IL-4诱导生成DC；用Ficoll分离脐血单个核细胞，2h贴壁后的细胞，以GM-CSF、IL-4和TNF-α诱导生成DC。观察细胞形态，流式细胞仪分析表面标志(CDla、CD80、HLA-DR)，并检测其刺激同种异体T细胞增殖的能力。结果 人脐血CD34^ 细胞和贴壁细胞体外诱导生成的细胞均具有典型的DC形态特征，表达高水平的DC分化抗原CDla，MHCⅡ类抗原递呈分子HLA—DR和CD80共刺激分子，并具有刺激同种异体T细胞增殖的能力。前者扩增DC的效率更高。结论 从人脐血CD34^ 细胞和贴壁细胞均可诱生出功能性DC，是DC理想的组织来源。     

人脐血不同前体细胞体外诱导分化为树突状细胞的研究

目的从人脐血不同的前体细胞体外诱导分化树突状细胞(DCs),并对生成的细胞进行鉴定,探讨脐血在DC体外大量扩增的组织来源作用.方法用免疫磁珠法分离人脐血CD34 细胞,以GM-CSF、TNF-α、FL、SCF、IL-4诱导生成DC;用Ficoll分离脐血单个核细胞,2h贴壁后的细胞,以GM-CSF、IL-4、TNF-α诱导生成DC.电镜观察细胞形态,流式细胞仪分析细胞表型,并检测其刺激同种T细胞增殖的能力.结果人脐血CD34 细胞和贴壁细胞体外诱导生成的DC均具有典型的DC 形态特征,表达高水平的DC特异性标志CDla,MHCⅡ类抗原递呈分子HLA-DR和CD80共刺激分子.两种方法获得的DC的形态、CDla表面抗原的表达没有显著的差异,两者均具有刺激同种T细胞增殖的能力,前者扩增DC的效率更高.结论从人脐血中CD34 细胞和贴壁细胞两种DC的前体细胞均可诱生出DC,脐带血是DC理想的组织来源.     

树突状细胞体外培养体系的优化

目的:探讨无血清专用培养基从人外周血单个核细胞(DC)分离培养树突状细胞的优化方法。方法:取正常成人新鲜血液,经密度梯度离心法,利用连续贴壁的方法获得单个核细胞,采用无血清培养基经人重组粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)、重组人白介素-4(rhIL-4)和肿瘤坏死因子-α(TNF-α)诱导产生成熟DC,倒置显微镜观察树突状细胞结构,流式细胞仪检测DC表面标志物表达水平。结果:连续贴壁法无血清专用培养基体外分离培养人外周血DC,所获得DC数量和纯度均都多于以往一次贴壁分离含10%胎牛血清RPMI培养基培养的DC。培养1周的DC高表达CD83、HLA-DR、CD80、CD86、CD83 HLA-DR 及CD80 CD86 。结论:采用无血清专用培养基连续贴壁法可分离培养数量较大、纯度较高的人外周血来源的DC。     

人外周血单核细胞体外诱导树突状细胞及鉴定

目的：建立从人外周血单核细胞体外诱导培养树突状细胞（dendritic cell,DC)的方法。方法：分离人外周血单个核细胞,以细胞因子粒细胞－巨噬细胞集落刺激因子（GM－CSF）,白细胞介素－4（IL－4）,肿瘤坏死因子α（TNF－α）诱导培养获得DC,电镜及共聚集显微镜观察其形态,流式细胞术检测细胞表面抗原,体外同种混合淋巴细胞反应检测DC刺激T细胞的增殖活性。结果：从正常外周血分离得到的单核细胞,体外经重且人GM－CSF,IL－4,TNF－α的共同诱导培养,得到大量成熟DC,形态学观察可见典型DC特征,荧光激少在细胞分离器（FACS）检测表明,诱导的DC高表达HLA－DR（96％）,CD83（94．2％）,CDla(94.2%)分子,同时也高表达CD40（97．7％）,CD80（98．2％）分子,同种混合淋巴细胞反应显示,诱导的DC具有很强的激发同种T细胞增殖的能力。结论：人外周血单核细胞体外经细胞因子贯序诱导培养,可以生成大量功能成熟的DC,为进一步开展DC的基础研究和临床应用提供了可能。     

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.