Reduced graft-versus-host disease-inducing capacity of T cells after activation, culturing, and magnetic cell sorting selection in an allogeneic bone marrow transplantation model in rats.

Graft-versus-host disease (GvHD), a major complication of allogeneic bone marrow transplantation, has been ascribed to mature T cells in the graft. Because T cells play an important role in engraftment of the bone marrow and decrease the probability of relapse of leukemia, a treatment strategy was developed to preserve the benefits of T cells in the graft and to control the severe complications of GvHD. This can be accomplished by the genetic modification of donor T cells with a suicide gene that allows their selective in vivo elimination and subsequently the abrogation of GvHD. For clinical benefit the alloreactivity of herpes simplex virus thymidine kinase (HSV-TK) gene-transduced T cells should be retained. Therefore, we investigated the influence of gene transduction and the selection procedure on T cells. We demonstrated that activation and culturing of T cells reduce their capacity to induce lethal GvHD in an allogeneic rat bone marrow transplantation model. Furthermore, positive immunomagnetic selection of gene-transduced T cells resulted in loss of the GvHD-inducing capacity of HSV-TK(+) T cells directly after MACS (magnetic cell sorting) selection; this loss could be recovered by a 1-day expansion of the selected T cells. No effect on alloreactivity was observed to be caused by the gene transduction procedure. Our study resulted in the development of an optimized culture and gene transduction protocol with preservation of T cell alloreactivity. Treatment of transplanted rats with ganciclovir resulted in a rapid reduction in the number of HSV-TK(+) T cells in the peripheral blood and in increased survival of the animals.     

Bioavailability and biological efficacy of a new oral formulation of salmon calcitonin in healthy volunteers.

Salmon calcitonin (SCT) is a well-tolerated peptide drug with a wide therapeutic margin and is administered parenterally for long-term treatments of bone diseases. Its clinical usefulness would be enhanced by the development of an orally active formulation. In this randomized crossover double-blinded phase I trial, controlled by both a placebo and a parenteral verum, we have tested a new oral formulation of SCT associated with a caprylic acid derivative as carrier. Eight healthy volunteers received single doses of 400, 800, and 1200 microg of SCT orally, a placebo, and a 10-microg (50 IU) SCT intravenous infusion. SCT was reliably absorbed from the oral formulation, with an absolute bioavailability of 0.5-1.4%, depending on the dose. It induced a marked, dose-dependent drop in blood and urine C-terminal telopeptide of type I collagen (CTX), a sensitive and specific bone resorption marker, with the effects of 1200 microg exceeding those of 10 microg intravenously. It also decreased blood calcium and phosphate, and increased the circulating levels of parathyroid hormone (PTH) and, transiently, the urinary excretion of calcium. It was well-tolerated, with some subjects presenting mild and transient nausea, abdominal cramps, diarrheic stools, and headaches. This study shows that oral delivery of SCT is feasible with reproducible absorption and systemic biological efficacy. Such an oral formulation could facilitate the use of SCT in the treatment of osteoporosis and other bone diseases.     

慢性乙型肝炎中可溶性白细胞介素2受体与其它血清学指标的关系

目的 :探讨血清可溶性白细胞介素 2受体 (sIL 2R)在慢性乙型病毒性肝炎 (慢乙肝 )中的发病机制及sIL 2R与肝纤维化指标、肝功能的相关关系。方法 :对 312例肝活检病人按组织病理学炎症和纤维化程度进行分级分期 ,同时检测了病人血清sIL 2R、肝功能及肝纤维化相关指标。结果 :血清sIL 2R均值较对照组明显增高 (P <0 .0 1) ,增幅随着肝脏病理炎症和纤维化程度的加重而逐渐加大 ,各组间比较差异有显著性 (P <0 .0 5或P <0 .0 1)。sIL 2R与血清白蛋白 (A)呈显著负相关 ,与血清脯氨酸肽酶 (PLD)、Ⅳ型胶原 (CⅣ )、层粘连蛋白 (LN)、透明质酸 (HA)、Ⅲ型前胶原氨基端肽 (PⅢNP)、肝功指标中的TB、DB、G、ALT、AST、AKP、γ GT呈显著正相关 (P <0 .0 5或P <0 .0 1)。结论 :慢乙肝病人血清sIL 2R水平与A呈显著负相关 ,与PLD、CⅣ、LN、HA、PⅢNP呈显著正相关 ;sIL 2R的血清水平能反映肝脏组织学病变的程度 ,可作为判断肝组织炎症和纤维化程度的参考指标。     

成人成骨细胞与珊瑚羟基磷灰石的体外生物相容性

目的：研究成人骨髓成骨细胞与珊瑚羟基磷灰石（carolline hydroxyapatite,CHA)在体外培养条件下的生物相容性。方法：抽取健康成人骨髓组织，置于含体积分数为10％胎牛血清的DMEM培养基中培养，传代后改用含地塞米松、β-甘油磷酸钠和维生素C的条件培养基培养，分为CHA复合细胞组和单纯细胞组，不同时间用倒置相关显微镜，HE染色光镜及扫描电镜观察，MTT法进行细胞增殖测定，并进行细胞微量蛋白含量和碱性磷酸酶的定量检测，结果：成人骨髓成骨细胞体外培养时复合或不复合CHA均生长良好，表现出典型成骨细胞的形态特征和生物学特性，CHA利于细胞的贴附，生长与增殖，并对细胞的功能无不良影响，结论：CHA是较理想的骨组织工程支架材料，成骨细胞复合CHA用于骨缺损的修复，具有广阔的临床应用前景。     

VEGF在膀胱移行细胞癌中的表达及意义

目的：探讨膀胱移行细胞癌中血管内皮生长因子（VEGF）的表达及其与临床病理指标的关系。方法：应用免疫组化方法对40例膀胱移行细胞癌及15例正常膀胱组织中VEGF进行检测。结果：正常膀胱移行上皮均为阴性表达；膀胱移行细胞癌中VEGF阳性表达24例，表达率为60％（24／40），明显高于正常膀胱组织（P＜0.01），其中VEGF表达阳性率随膀胱癌病理分级、临床分期上升而升高，表达强度也随之增强（P＜0.01）。结论：VEGF表达对膀胱移行细胞癌生物学行为有重要影响，VEGF有可能可作为判断膀胱癌生物学行为、转移潜能及预后指标。     

硬膜外穿刺针外套管在腹腔镜小儿斜疝高位结扎术中的应用

目的探讨硬膜外穿刺针外套管在腹腔镜小儿斜疝疝囊高位结扎中的应用价值。方法2003年4月-2006年5月,我院对230例小儿斜疝手术应用硬膜外穿刺针外套管代替雪橇钩针实施腹腔镜疝内环口荷包高位结扎术。结果230例手术均获得成功,手术时间：单侧161例8-10 min,嵌顿疝39例15-20 min,双侧30例16-20 min;术后未见有阴囊水肿及睾血疼痛及缺血坏死。术后随访2-6个月111例,6-12个月52例,12-24个月37例,24-36个月30例,无复发。结论硬膜外刺针外套管可替代小儿疝囊结扎专用的雪橇钩针,简便,经济,适合在所有基层医院推广。     

p53,p16,PCNA蛋白在食管癌中的表达及其临床意义

目的探讨p53,p16,PCNA蛋白在食管癌中的表达及临床意义。方法用免疫组织化学染色SP法对62例食管癌标本进行p53,p16,PCNA蛋白测定。结果62例食管癌中,p53,PCNA蛋白阳性表达均为71.0%(44/62),p16缺失率48.4%(30/62)。p16缺失与肿瘤浸润深度、淋巴结转移密切相关(P<0.05、P<0.01)。而p53,PCNA蛋白同时表达56.5%(35/62),亦与肿瘤浸润深度、淋巴结转移密切相关(P<0.05、P<0.01)。结论食管癌p53,PCNA蛋白同时表达及p16缺失可视为危险预后因素。     

保留盆腔植物神经对老年男性患者术后性功能及排尿功能的影响

目的:探讨保留盆腔植物神经的直肠癌手术对老年男性患者性功能及排尿功能的影响。方法:1999~2005年因直肠癌手术的170例老年男性患者分为PANP组(含规范的TME手术方法)和非PANP(TME规范情况不详),对两组术后性功能、排尿功能和局部复发率进行回顾性总结、比较。结果:PANP组性功能与排尿功能明显优于非PANP组,但统计学无显著差异(P<0.05)。而PANP组局部复发率低于非PANP组(P>0.05)。结论:老年直肠癌患者应重视性功能和排尿功能的保护。     

术后镇痛10年后的质量控制探讨：1168例回顾性研究

目的 了解术后镇痛的现状，探讨术后镇痛的质量控制问题。方法 回顾近3个月1168例术后镇痛所采用的镇痛方法和预防术后恶心呕吐的方法，了解镇痛效果和副作用的发生情况，并分析影响镇痛效果和副作用发生率的有关因素，患者的满意度情况。结果 90％采用了连续硬膜外镇痛法，配方主要是小剂量吗啡复合低浓度布比卡因或罗比卡因，静脉镇痛主要采用吗啡复合氯诺昔康。预防术后恶心呕吐（PONV）的方法有5-HT3受体拮抗剂（昂丹司琼或阿扎司琼），氟哌啶，地塞米松等，单用或复合应用。连续硬膜外镇痛需要补救镇痛者约5％，静脉镇痛补救镇痛者约10％。PONV总体发生率低，妇科开腹手术低至20％，女性非妇科手术低至10％，剖宫产术低至1％，使用氟哌啶使妇科手术PONV降低约50％。1例高龄患者出现血氧饱和度下降至82％，经停药、吸氧等处理好转。硬膜外导管脱落0．5％。瘙痒、头晕等并发症小于5％。通过补救镇痛，及时处理并发症，患者满意度在90％以上。结论 硬膜外镇痛方法效果优于静脉镇痛方法，适度镇痛，减少镇痛药量，增加安全性，重视PONV的预防，及时处理镇痛不足和副作用，从而提高患者的满意度，是术后镇痛质量控制的可行的理念和方法。     

Transforming Growth Factor-β1 Induces Apoptosis through Down-Regulation of c-mycGene and Overexpression of p27Protein in Cervical Carcinoma

Transforming growth factor-β1 (TGF-β1) is known to be a potent growth inhibitor for many cell types, including most epithelial cells. In skin keratinocytes, TGF-β1 has been shown to inhibit growth and to rapidly reduce c-mycexpression. However, the molecular mechanism of TGF-β1 action on cell growth of cervical carcinoma has not yet been elucidated. We thus assessed the effect of TGF-β1 on the growth of cervical carcinoma cell lines. Two cervical squamous carcinoma cell lines, CUMC-3 and CUMC-6, were incubated with varying concentrations of TGF-β1, and growth inhibition was evaluated with tetrazolium-based colorimetric assay. After culture in TGF-β1 for 24 h, inhibition of growth was detected in a dose-dependent manner at concentrations of 0.1–10 ng/ml in both cell lines. This effect of TGF-β1 on cultured carcinoma cells was associated with apoptotic process including oligonucleosomal ladder DNA and apoptotic body formations. Northern blot analysis revealed c-mycmRNA expression was suppressed by 10 ng/ml of TGF-β1 following 3 h of treatment in both cell lines. Western blot analysis showed that the level of p27^Kip1protein was increased after TGF-β1 treatment in both cell lines. These results suggest that the mechanisms by which TGF-β1 inhibits the growth of cervical carcinoma are complex and may include effects on down-regulation of c-mycgene, and overexpression of p27^Kip1protein.