儿童脂肪干细胞与多种尿道生物支架相容性的对比研究

目的:比较儿童脂肪干细胞与多种尿道生物支架的相容性。方法:将儿童脂肪干细胞与膀胱脱细胞基质、小肠粘膜下层组织、脱细胞尿道海绵体基质分别体外复合培养,运用倒置相差显微镜观察干细胞在这3种支架上的生长、增殖情况,并且测定细胞活性与细胞黏附率。结果:脂肪干细胞能够在3种支架上生长良好,生物活性无统计学差异(P>0.05),膀胱脱细胞基质细胞黏附率高于脱细胞尿道海绵体基质(P<0.05)。结论:脂肪干细胞与3种生物支架均有较好的相容性,膀胱脱细胞基质或是最好的选择。     

表面修饰化材料对颌下腺种子细胞生长影响的研究

目的 探讨应用表面修饰技术处理的生物支架材料与颌下腺种子细胞复合培养研究对种子细胞支架材料上的附着、生长和分泌功能的影响.方法 选用明胶海绵36块,均分为3组.A组:明胶海绵材料与鼠颌下腺细胞复合培养物;B组:层黏连蛋白(laminin,LN)表面修饰的明胶海绵材料与鼠颌下腺细胞复合培养物;C组:转化生长因子(TGF-β3)表面修饰的明胶海绵材料与鼠颌下腺细胞复合培养物.各组在体外进行复合培养,3、7和15 d各时间点行组织学观察和扫描电镜观察,15 d行免疫组织化学鉴定细胞来源,测定上清淀粉酶含量并进行比较.结果 组织学和扫描电镜观察见培养第3天各组细胞分散于材料表面,无细胞突起形成;第7天见B组细胞数量明显多于A组和C组,细胞突起形成并锚定于胶原海绵表面;第15天,B组细胞数量仍多于A组和C组,并可见细胞之间有触突联系和腺管样结构形成.免疫组织化学染色观察复合培养的颌下腺细胞Brdu呈强阳性.随着接种后培养时间的延长,各组颌下腺细胞分泌的淀粉酶含量均有不同程度的增加.15d,C组淀粉酶含量明显高于A组和B组(P＜0.05).结论 应用细胞外基质和细胞因子成分对生物支架材料进行表面修饰,可促进颌下腺种子细胞在材料上的附着、增殖和分泌,有利于组织工程化颌下腺研究的进一步成功.     

诱导向尿路上皮分化的脂肪干细胞与膀胱脱细胞基质支架的生物相容性研究

目的 探讨脂肪干细胞向尿路上皮细胞分化的可行性,并将诱导后细胞与膀胱脱细胞基质复合培养,评价细胞与支架材料的生物相容性,为进一步构建组织工程化组织提供实验基础。 方法 取6只8周龄雄性新西兰大白兔的腹股沟脂肪组织和膀胱组织,通过胶原酶消化法分离脂肪干细胞(adipose-derived stem cells,ADSCs),同时通过胰蛋白酶消化的方法获取尿路上皮细胞,流式细胞鉴定后将第3代脂肪干细胞与尿路上皮细胞间接共培养2周,并对分化后细胞进行表型鉴定。随后将分化后细胞种植在膀胱脱细胞基质上,通过扫描电镜、组织学切片观察诱导后细胞在膀胱脱细胞基质上的生长情况。 结果 脂肪干细胞培养成功并在体外扩增,流式术检测表面抗原CD分子,细胞高表达CD29(99.6%)、CD44(98.2%);低表达CD31(1.9%)及CD45(1.6%)。通过间接共培养诱导分化后,细胞免疫荧光、Western blotting检测到尿路上皮标志物UPIa的表达,而未诱导的脂肪干细胞无表达。扫描电镜提示诱导后细胞在膀胱脱细胞基质上生长,黏附较好。组织学检查可见膀胱脱细胞基质表面细胞平铺生长。 结论 脂肪干细胞向尿路上皮诱导分化后可作为泌尿系组织工程的种子细胞,可能是尿路上皮细胞之外的又一新选择。膀胱脱细胞基质能支持诱导后的脂肪干细胞良好的生长,该支架可作为载体材料用于泌尿系组织工程的修复与重建。      

脱钙骨基质支架构建组织工程骨的实验研究

目的以同种异体骨脱钙骨基质(demineralized bone matrix,DBM)为支架材料,复合体外诱导培养的人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs),构建组织工程骨并通过裸鼠皮下异位成骨实验及裸鼠皮下致瘤性实验验证其成骨效果及安全性.方法贴壁法培养hMSCs,体外诱导培养扩增,以1.857×106/ml的密度与DBM复合构建组织工程骨,体外培养,并在扫描电镜下观察细胞与材料复合情况,进行组织学观察.同时进行了裸鼠皮下异位成骨实验.结果细胞与支架复合后3 d,细胞与DBM表面及孔隙可实现良好复合;复合后5 d,扫描电镜观察到细胞基质分泌旺盛,充满支架孔隙.裸鼠皮下成骨实验证明其成骨效果良好.结论以本实验中使用的细胞培养方法扩增、诱导分化的种子细胞生物安全性好,自制的DBM具有良好的生物相容性,可为种子细胞生长提供较好的三维空间,按照标准化工艺流程制备的组织工程骨安全、有效.     

胎兔成骨细胞与复合肝细胞生长因子/异种脱蛋白松质骨的体外培养

目的:探讨肝细胞生长因子(HGF)对体外培养的胎兔成骨细胞在异种脱蛋白松质骨(DPB)支架上黏附行为及增殖和分化功能的影响.方法:成骨细胞从胎兔中分离,实验组为HGF/DPB与成骨细胞复合培养,对照组为DPB与成骨细胞常规培养.通过电镜观察成骨细胞在HGF/DPB表面生长与附着状态,同时检测细胞增殖状况和碱性磷酸酶(alkalinephosphatase,ALP)活性等,评价成骨细胞生长情况和细胞活性.结果:成骨细胞在复合肝细胞生长因子脱蛋白骨外表面及孔隙内表面均可贴附生长,增殖、分化良好.HGF/DPB对成骨细胞的ALP活性有明显的促进作用,并能促进成骨细胞的增殖.结论:HGF/DPB具有良好的生物相容性和骨诱导作用,可作为骨组织工程备选的复合支架材料.     

四种显微技术观测大鼠颌下腺细胞与丝素-壳聚糖体外共培养的形态学特点

目的 采用倒置显微镜、扫描电镜(scanning electron microscopy,SEM)、荧光显微镜和激光共聚焦显微镜(( laser scanning confocal microscopy,LSCM))技术对大鼠颌下腺细胞(rat submandibular gland cells,RSMGs)与丝素-壳聚糖( silk fibroin-chitosan,SFCs)的体外复合培养进行形态学观察.为观测、评估种子细胞在三维支架的内部生长情况提供技术支持.方法 取0～8d龄SD大鼠的颌下腺,对大鼠颌下腺细胞进行原代培养、分离纯化并传代;用抗细胞角蛋白单克隆抗体( CK8)及淀粉酶抗体的免疫细胞化学染色鉴定细胞来源.选取传至第二代的对数生长期的RSMGs作为种子细胞,选取SFCs共混膜(5×5×2)mm作为支架材料构建组织工程化涎腺样结构.将种子细胞与支架材料复合培养并分别于倒置显微镜、SEM、荧光显微镜和LSCM下观察二者复合生长情况.结果 倒置显微镜可以直接观察活细胞与支架复合生长情况,方法简单易行.SEM可以较精确的展示细胞支架复合生长的表面超微结构.经过荧光染料的着色,荧光显微镜和LSCM都可以观察到支架上锚定的种子细胞.荧光显微镜可见细胞核的荧光信号均匀的分布在支架孔隙内.LSCM通过层扫描及三维重建技术对较厚的标本获取图像;并可以通过旋转图像,从不同角度观察细胞支架复合物的三维剖面或整体结构,得到更为准确的定位信息.结论 四种显微技术均可应用于RSMGs与SFCs体外共培养的形态学观测.LSCM的三维重建技术结合荧光染料标记可以较好地获得RSMGs与SFCs复合生长的情况,有着较广泛的应用价值.     

家猪脱髓核细胞基质的制备及其生物相容性评价

目的:制备家猪脱髓核细胞基质并评价其生物相容性。方法:分离家猪脊柱髓核组织,并逐步经过胰酶、核酶及TritonX-100处理,得到脱髓核细胞基质,DAPI染色观察脱细胞效果及扫描电镜观察脱髓核细胞基质的物理形态特点,将脱髓核细胞基质溶于醋酸,制备成脱细胞基质膜,将大鼠骨髓间充质干细胞接种于基质膜上培养。在第1～5天采用CCK-8检测骨髓间充质干细胞在脱髓核细胞基质膜上的增殖情况,并计算细胞相对增殖率(RGR)来评价其生物相容性。结果:制备出的家猪脱髓核细胞基质肉眼呈白色糜状,DAPI染色发现无细胞残留。扫描电镜下观察发现,脱髓核细胞基质由无序排列的胶原纤维组成。通过CCK-8检测,5 d内细胞毒性分级在0级与2级之间。结论:家猪正常髓核组织能够进行脱细胞处理,且脱细胞后由无序的胶原纤维组成。脱髓核细胞基质具有很好的生物相容性,符合生物工程支架选材标准。     

兔骨髓源成骨细胞与多孔支架复合的实验研究

目的通过对兔骨髓基质干细胞的培养及诱导分化,并将细胞与新型复合型生物多孔支架联合培养,观察兔骨髓源成骨细胞在多孔支架表面的黏附及生长情况。方法兔骨髓基质干细胞经诱导分化为成骨细胞。将成骨细胞与处理过的多孔支架复合,在扫描电镜下观察细胞在支架表面的贴附情况。结果细胞接种后,在材料表面生长形态正常,属性与特征明显。结论新型复合型生物多孔支架与兔骨髓源成骨细胞具有良好的结合能力,可以进一步进行体内试验。     

组织工程化颌下腺细胞与支架材料复合培养最佳细胞浓度的实验研究

目的 :研究颌下腺细胞与支架材料体外复合培养中 ,细胞接种浓度对复合培养物的影响。方法 :体外原代培养SD鼠颌下腺细胞 ,并传代至第 2代 ,细胞计数 ,按细胞浓度为 10 ,2 0 ,30 ,4 0 ,5 0 ,6 0 ,70× 10 6/ml接种于 5mm× 5mm× 2mm胶原海绵表面 ,并置于 2 4孔培养板内 ,形成颌下腺细胞 胶原海绵复合物。继续培养 ,于接种细胞后 1,3,5 ,7d取材 ;HE染色、免疫组化、扫描电镜观察及细胞计数检测等 ,评价颌下腺细胞 -胶原海绵复合物形成的最佳细胞接种浓度。结果 :5 0 ,6 0 ,70× 10 6/ml细胞浓度的复合物中颌下腺细胞在支架材料上黏附、伸展及生长良好。 10 ,2 0 ,30 ,4 0× 10 6/ml细胞浓度的复合物中颌下腺细胞在支架材料上增殖分化及生长较差。细胞计数显示接种 7d时 ,各组材料细胞数均只有 10 4/ml数量级 ,5 0 ,6 0 ,70× 10 6/ml接种浓度时 ,细胞粘附数量是 10 ,2 0 ,30 ,4 0× 10 6/ml的 2倍。结论 :颌下腺细胞与胶原海绵复合培养最佳接种细胞浓度为 5 0× 10 6/ml。与其它组织细胞复合培养的最佳接种浓度相似     

猪脱细胞基质矿化支架材料的生物相容性及降解特性

目的 探讨猪脱细胞基质矿化支架材料的生物相容性及体外降解特性.方法 体内实验将猪脱细胞基质矿化支架材料(HA-PADM)、猪脱细胞基质支架材料(PADM)、明胶海绵分别埋植在12只新西兰大白兔脊柱两侧肌肉内,并且于观察时间(7、14、21、28 d)取出,组织病理学切片HE染色后观察组织的炎症反应,同时于术前、植入第2天、第4天兔耳缘静脉抽血行血常规检查;体外实验将HA-PADM、PADM、明胶海绵浸泡于细胞培养液(DMEM)中,于各个时间点(7、14、21、28 d)称重研究材料的体外降解情况.结果 各时间点实验动物的血常规检测结果无显著性差异(P＞0.05);三种支架材料周围肌肉组织无脓肿形成和组织坏死发生;此种支架材料的降解特性符合牙周组织工程的要求.结论 猪脱细胞基质矿化支架材料生物相容性良好,降解时间适宜,是一种应用前景广阔的牙周组织工程支架材料.     

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.