宫颈癌组织HPV58的检测及其E7基因的克隆和表达

目的　检测宫颈癌组织中人乳头瘤病毒 5 8型 (HPV5 8)并克隆表达其E7基因。方法采用GP5 /GP6 引物系统扩增 5 8例宫颈癌组织 ,将荧光偏振 (FP)检测技术与模板指导的末端延伸反应 (TDI FP)结合 ,检测HPV5 8,确定HPV5 8感染阳性宫颈癌组织。用特异引物从HPV5 8阳性标本中扩增HPV5 8早期表达蛋白E7基因 ,将其连入pGEM TEasy载体 ,获得克隆重组体HPV5 8E7 pGEM T ,并测序验证。将E7基因与pRSET A融合表达载体连接 ,获得E7表达重组体pRSET 5 8E7,转化大肠杆菌BL2 1(DE3) ,并用IPTG诱导表达。结果　 5 8例宫颈癌标本中 ,HPV5 8阳性 10例 ,占 5 2例HPV阳性标本的 19.2 %。从其中扩增到了HPV5 8E7基因并构建了其克隆和表达重组体。其表达重组体经IPTG诱导后 ,可表达Mr16× 10 3 的HPV5 8E7His6融合蛋白 ,表达量占菌体蛋白的 30 %。结论　欧美国家少见的高危HPV5 8在中国陕西人宫颈癌组织中并不少见。HPV5 8E7重组表达体可在大肠杆菌中高效表达 ,为HPV5 8相关肿瘤诊断试剂及疫苗的研制奠定了初步基础     

High Bioavailability of α-Tocopherol Loaded into Poly (DL-Lactic-co-Glycolic Acid) Microspheres in Apolipoprotein B Knockout Mice

PURPOSE: To assess the potential clinical value of alpha-tocopherol-loaded poly (DL-lactic-co-glycolic acid) (PLGA) microspheres, we examined the disposition kinetics of alpha-tocopherol after administration of the microspheres to apolipoprotein B (apo B) knockout mice as a model of abetalipoproteinemia. METHODS: PLGA microspheres containing alpha-tocopherol were prepared by a solvent-evaporation method. The concentration of alpha-tocopherol was measured by gas chromatography-mass spectrometry. RESULTS: The mean value of particle size of alpha-tocopherol-loaded PLGA microspheres was 108 microm. The loading and the trapping efficiency of alpha-tocopherol in PLGA microspheres were 20.8% and 86.6%, respectively. When alpha-tocopherol solution (25 mg/kg) was subcutaneously administered to apob (+/+) and apob (+/-) mice, the plasma concentrations of alpha-tocopherol reached a peak at 6 h and decreased to the endogenous level within 4 days in both types of mice. However, the area under the plasma concentration-time curve (AUC) of apob (+/-) mice was significantly smaller than that in the case of apob (+/+) mice. When alpha-tocopherol-loaded PLGA microspheres (100 mg/kg) were subcutaneously administered, the plasma concentrations of alpha-tocopherol increased slowly and remained about 2-fold higher than the endogenous level at 5 to 10 days after administration in both types of mice, and there was no significant difference between the AUC values. CONCLUSIONS: The PLGA microsphere preparation of alpha-tocopherol is expected to be a very useful drug delivery system in vitamin E supplementation therapy for abetalipoproteinemia.     

Organ culture of the postnatal mouse crista ampullaris part I

Summary The 2 day post partum mouse Crista ampullaris was cultured in vitro for 7 days. The differentiation into secretory and sensory areas was preserved. The ultrastructure of the dark cells of the secretory region was well maintained. The hair cell morphologically was considerably changed though the hair cells themselves survived 7 days in culture.Supported by grants from Karolinska Institutet, and Swedish Medical Research Council (Grant No. 12X-720)     

Promoter methylation of the RGC32 gene in nonsmall cell lung cancer

BACKGROUND:

Lung cancer is the leading cause of cancer‐related deaths worldwide. Epigenetic inactivation of certain genes by aberrant promoter methylation is recognized as a crucial component in the initiation and progression of lung cancer. Response gene to complement 32 (RGC32) has been identified as a cell cycle regulator induced by activation of complements; however, its role in carcinogenesis is still controversial.

METHODS:

The authors examined the methylation status in the promoter region of RGC32 gene in nonsmall cell lung cancers (NSCLCs) using a methylation‐specific PCR and correlated the results with clinicopathological features.

RESULTS:

RGC32 methylation was found in 45 of 173 NSCLCs (26.0%) and was related to the gene expression. RGC32 methylation was more frequent in females than in males (P<0.05). RGC32 methylation was not significantly associated with the prognosis of patients; however, when the patients were categorized by TP53 mutational status, the effect of RGC32 methylation on prognosis was significantly different between those with and without TP53 mutations (P = .005 [test for homogeneity]). Specifically, RGC32 methylation was associated with significantly worse survival in the cases with wild‐type TP53, whereas it exhibited a better survival outcome in the cases with TP53 mutations.

CONCLUSIONS:

The current findings suggest that methylation‐associated down‐regulation of RGC32 plays an important role in the pathogenesis of NSCLC, particularly in females. However, further studies with a large number of cases are needed to confirm the authors' findings. Cancer 2011. © 2010 American Cancer Society.     

Cryopreserved arterial allografts for in situ reconstruction of abdominal aortic native or secondary graft infection

Objective

The objective of this study was to evaluate the early and long-term outcome of cryopreserved arterial allografts (CAAs) used for in situ reconstruction of abdominal aortic native or secondary graft infection and to identify predictors of mortality.

Biomechanical analysis of supra-acetabular insufficiency fracture using finite element analysis

Background

Supra-acetabular insufficiency fractures (SAIFs) occur in the upper acetabulum and are rare compared with insufficiency sacral, femoral head, or ischial fractures. However, SAIFs are known to occur in low grade trauma, and the underlying mechanism is still remained unclear.