反相高效液相色谱法测定发酵液中梅岭霉素

目的:建立了发酵液中梅岭霉素的定量测定方法。方法:反相高效液相色谱法,Hypersil-C_(18)柱(4 mm×125mm,5μm),流动相为乙腈-水(78:22),流速1.2mL·min~(-1),检测波长238 nm,室温测定。结果:在1O.2～163.2μg·mL~(-1)范围内梅岭霉素浓度与峰面积呈良好线性关系,r=0.999 7,回归方程为Y=25.44X+2.72。样品的日间 RSD≤1.8％,高中低浓度样品的日内RSD≤2.5％(n=8)。平均回收率为96.9％(n=8,RSD<3.0％)。结论:本方法快速、准确、简便、稳定,为定量分析提供了可靠的依据。     

Time-dependent astroglial changes after gamma knife radiosurgery in the rat forebrain

OBJECTIVE: Using an experimental rat model and a clinically relevant treatment dose, we performed gamma knife radiosurgery to define the hyperacute radiation effects in normal rat forebrain, the time dependence of the astrocytic reaction, and the participation of astrocytes in the healing process after single-dose gamma radiation injuries. METHODS: Seventy-one rats underwent radiosurgical treatment (4-mm collimator) of the caudate-putamen nucleus (single-fraction maximal dose of 100 Gy) and were killed at times ranging from 3 hours to 90 days. Serial cryostat brain sections were processed with the immunohistochemical avidin-biotin complex technique, using anti-glial fibrillary acidic protein as the primary antibody (to identify astrocytes). RESULTS: Vascular changes, including endothelial hyperplasia and vessel wall thickening, were identified as the earliest postradiation manifestations and continued throughout the observation period. Astrocytes reacted to the radiation injury with hyperplasia and hypertrophy. At earlier time points (3-24 h), proliferation was the predominant reaction. The expression of glial fibrillary acidic protein in the proliferating and hypertrophic astrocytes formed an initial peak in the adjacent corpus callosum 3 days after radiosurgery and peaked within the target site between 14 and 30 days. Astrocytic proliferation and hypertrophy were also observed in distant cortices (frontal, parietal, insular, and piriform cortices) and in the hippocampus. No necrosis was observed less than 30 days after irradiation. By Day 90, necrotic lesions with a mean diameter of 4 mm were identified, with glial scar at their peripheries. Astrocytic morphological features varied according to the distance from the necrosis. The irradiated side contained more glial fibrillary acidic protein-containing cells than did the nonirradiated contralateral side. CONCLUSION: During the early phase after radiation, vasculopathy was the first morphological change and may serve as the initiating factor for subsequent changes. Reactive astrocytes appeared not only at the target site but also in the surrounding regions; the severity of injury was determined by the distance from the target.     

旋转手法对椎动脉血流平均速度的影响

目的　观察手法治疗椎动脉型颈椎病的经颅多普勒 (TCD)变化 ,从血流动力学角度探讨手法治疗的作用机制。方法　将 82例椎动脉型颈椎病患者随机分成手法治疗组 4 0例与常规治疗组 4 2例 ,治疗前及治疗后检查TCD ,观察手法治疗对椎动脉型颈椎病患者的血流平均速度的影响。结果　① 82例患者治疗前椎基底动脉血流平均速度均明显低于正常参考值范围(P <0 .0 5 )。TCD异常率为 76 % ( 6 2 /82 )。②治疗后两组椎基底动脉血流速度均有改善。手法治疗组比常规治疗组椎动脉血流速度改善更明显 (P <0 .0 5 )。结论　手法治疗可以改善椎动脉型颈椎病椎基底动脉的血流速度。     

Congenital bladder diverticulum and Ehlers-Danlos syndrome: an unusual association

The existence of a vesical diverticulum in the context of a congenital connective tissue disorder such as Ehlers-Danlos syndrome led us to consider the possibility of a relationship. Four types of diverticula can be found in the literature: congenital, acquired, iatrogenic and syndrome-associated. Within the later, Ehlers-Danlos syndromes type IV and IX, even type V, are associated to the existence of vesical diverticula. The potential spontaneous rupture of the diverticulum is a typical feature, as well as post-surgery relapse. The attitude towards such diverticula should be one of watchful waiting, and simple, plasty-free diverticulectomy on the bladder's neck is indicated when performing a surgical procedure.     

Glandular cystic cystitis

Cystic-glandular cystitis is considered as part of the urothelial pre-neoplastic proliferative abnormalities. This group includes atypical hyperplasia. Von Brunn's nidus, and cystitis cystica. They are a consequence of the changes experienced at the urothelium level in response to inflammation, irritation or carcinogens. Diagnosis is mainly based in the pathoanatomical study of the biopsy obtained following endoscopic resection. The signs and symptoms it presents are varied and show a clear relationship to distribution and extension of cysts. Treatment is based in the removal of irritative factors. Cystectomy with urinary by-pass may be necessary if required by clinical evolution.     

肺癌患者血清PGE2水平及其对IL-2/sIL-2R的影响

目的 :探讨肺癌患者围手术期血清PGE2 水平的变化及其对IL 2 /sIL 2R的影响。方法 :采用放射免疫检测法 ( 12 5I RIA以及ELISA试剂盒 )分别检测 43例肺癌患者手术前、后的外周静脉血清PGE2 水平和IL 2 /sIL 2R ,术中抽取肿瘤回流静脉血用于检测PGE2 水平。结果 :肺癌患者外周静脉血存在较高的PGE2 ( 78 81± 3 2 15 )pg/mL ,与肿瘤回流静脉血中的PGE2 ( 13 1 0 7± 3 6 41)pg/mL呈显著相关 ,r =-0 3 15 ,P <0 0 1,术后其PGE2 显著下降 ;手术前后其IL 2 /sIL 2R处于相对较低水平 ,IL 2与PGE2 呈相关关系 ,而与sIL 2R无明显相关。术后IL 2有所上升 ( 15 3 83± 5 3 3 9)pg/mL ,但仍低于正常对照组 ( 2 3 0 2 9± 5 3 3 9)pg/mL ,P <0 0 5。结论 :肺癌患者存在着高水平的PGE2 ,是由肺部肿瘤引起的 ,对IL 2有一定的抑制作用。     

载体介导的RNA干扰技术抑制肝癌耐药细胞MDR1表达的研究

目的 :构建含多药耐药基因MDR1的短发夹状RNA(shorthairpinRNA ,shRNA)表达质粒 ,观察对肝癌耐药细胞Bel 740 2 /R的MDR1mRNA的抑制作用。方法 :利用分子克隆技术 ,将含MDR1的双链DNA ,与经双酶切后的载体PGE 1连接 ,构建pshRNA MDR1重组质粒 ,在脂质体的介导下转染肝癌耐药细胞株Bel 740 2 /R ;RT PCR分析MDR1mRNA的表达 ,MTT法检测细胞对药物的敏感性 ,流式细胞仪检测细胞内罗丹明 12 3 (Rh12 3 )的潴留和P gp的表达。结果 :PCR和DNA测序证实了表达质粒构建成功 ,并能明显地抑制MDR1mRNA的表达 ;对盐酸表柔比星和顺铂的半数抑制浓度IC50明显降低 ,P <0 0 5 ;P gp的表达阳性率降低了 5 7 3 % ;细胞内Rh12 3的浓度显著增高 ,P <0 0 5。结论 :构建的pshRNA MDR1表达质粒能有效地降低肝癌耐药细胞MDR1mRNA和P gp的表达。     

人类α型叶酸受体基因的克隆及其真核表达载体的构建

目的 探讨如何以人类α型叶酸受体(FOLR-1)基因为基础构建核酸疫苗。方法 应用RT-PCR技术，从人类卵巢癌细胞系-SKOV3细胞中扩增FOLR-1基因，插入克隆载体pGEM-T Easy，经DNA自动测序仪测序证实后，以亚克隆法构建于真核表达载体pcDNA3．1( )，并使用限制性内切酶酶切鉴定。结果 从卵巢癌细胞系SKOV3中成功扩增出FOLR-1基因，并克隆人pcDNA3．1( )载体。结论 成功构建FOLR-1的重组克隆及真核表达载体，为今后利用FOLR-1进行卵巢上皮性肿瘤的免疫及导向治疗研究打下了基础。     

梅岭霉素发酵过程参数相关分析和优化

对新型微生物农药梅岭霉素的发酵过程进行研究。通过对发酵过程表征细胞代谢和工程特性的特征参数：氧消耗速率、二氧化碳生成速率、呼吸商、氧传递系数等进行实时在线检测，并利用参数相关分析的方法。对原有工艺不断调整，实现了梅岭霉素发酵过程的初步优化，使发酵效价从原来的150u／ml提高到520u／ml。