Objective To observe the phenotypic changes between renal tubular epithelial cells and mesenchymal cells in rat tubulointerstitial fibrosis(TIF) .Methods The renal tubulointerstitial fibrosis models of Wistar rats were established by gavage with excessive adenine. The morphological changes of model kidney were observed by light microscopy, electron microscopy, polarizing microscopy. The?-smooth muscle actin(a-SMA), vimentin, and keratin were examined by immunohistochemistry staining. While the proliferating cell nuclear antigen (PCNA) was also assessed. Results There were many 2, 8-dioxyadenine crystals derived from the metabolite of adenine to deposit in the renal tubules, which induced the injury. Tubular epithelial cells degeneration and regeneration, loss of renal tubules, mononuclear cells infiltration in renal interstitium and the accumulation of extracellular matrix protein were observed. Both ?-SMA and vimentin were positive in some of renal tubules. And consecutive sections of immunohistochemical staining showed that the area of distribution of a-SMA positive cells was almost the same area as that of vimentin positive cells. Polarizing microscopy results showed that there was a mass of interstitial collagen ( Ⅲ and Ⅰ) accumulation. And electron microscopy examination proved that epithelial cells invaded into the renal interstitium. Conclusion There is tubular epithelial cells-myofibroblasts transdifferentiation in the process of TIF, and which plays a key role in the accumulation of extracellular matrix.

Objective To explore the clinicopathological features of IgA nephrolpathy associated with malignant hypertension (IgAN-MHT) and to analyze their correlation with renal vascular lesions. Methods Twenty-nine patients of IgAN-MHT were screened from 2000 biopsy-proven eases with primary IgA nephropathy (IgAN) in our department from April 1997 to May 2007. Data of clinicopathology and follow-up of these 29 patients were collected. Semi- quantitative analysis was performed to evaluate the pathological changes. Inner lumen, outer lumen, intimal thickness, tunica media-to-internal lumen ratio of 436 arterioles, 124 interlobular arteries and 5 arcuate arteries were measured. The primary endpeint was the composite of a doubling of serum creatinine level and ESRD. Correlations of renal vascular lesions with clinical manifestation, pathological change and prognosis were examined by Spearman and Cox methods. Results 1.5% of all the IgAN patients presented malignant hypertension. The common clinical features were renal failure (100%), hyperurieacidemia (62.7%) and hypertriglyceridemia (51.7%). The average amount of urine protein excretion was 2.8 g/d. The common pathological changes were moderate mesangial proliferation, severe global sclerosis, severe interstitial inflammation and severe interstitial- tubular fibrosis. The small arteries (arcuate arteries and interlobular arteries) and arterioles (afferent arterioles) were both involved in IgAN-MHT. The characteristic lesions of intrarenal arteries included vascular occlusion, media thickening, proliferative endarteritis (onionskin lesion, musculomucoid intimal hyperplasia), hyaline arteriosclerosis, but mainly vascular occlusion (86.2%). The arteriole lesion was negatively correlated with age and total protein level; vascular occlusion was positively correlated with uric acid level. The average foUow-up period was 21.1 months. Forteen patients reached the endpoint. The arteriole lesion was the main independent risk factor for the progression of IgAN-MHT (RR=10.21, 95%CI=1.16～89.67). Conclusions The main clinical feature of IgAN-MHT is renal failure. The main histological feature of intrarenal vascular lesions is occludes arterioles. Arteriole lesion is the main independent risk factor for the progression of IgAN-MHT.

Objective To observe the early changes of related indexes after high dose of 60Co γ-ray irradiation on rhesus monkey hematopoietic system.Methods A total of 33 rhesus monkeys were randomly divided into normal control and different irradiation control group,rhesus monkeys in irradiation control group were given different doses(4,8,12 Gy) irradiation to establish acute radiation sickness(ARS) models.XE-2100 automatic blood cell analyzer detected the peripheral blood before and after the irradiation of 3,6,9,12,24,48,80 h.The rhesus monkeys were sacrificed to have a observation of sternum pathological changes at 6,48 and 80 h after 4,8,12 Gy 60Co γ-ray irradiation.Results The number of white blood cell in peripheral blood of the rhesus monkeys after 4 and 8 Gy 60Co γ-ray irradiation were lower than that before irradiation at 3 h after irradiation,as was significant increased at 6 h after irradiation,the highest values were 136.04%.and 221.38% after 9 h(with before irradiation values was 100.00%,the same below),become obviously drooped from 12 h after irradiation,show clearly temporary peak.But the number of white blood cell after 12 Gy 60Co γ-ray irradiation was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood neutrophile count was significant increased at 6 h after irradiation,at the highest of 9 h,become obviously drooped from 12 h after irradiation.Peripheral blood lymphocyte count fell sharply after irradiation,3 h detection value was only 12.02%-25.04% of before irradiation.Sternal bone marrow nucleated cell number decreased sharply after irradiation,the more irradiation dose,the less residual hematopoietic cells.Conclusion In the early stage of BM-ARS,temporary peaktime node of the white blood cell and neutrophil count could be regarded as the best delivery time of hematopoietic cytokine therapy.

Objective: To study effects of cyclin D1 overexpression on the proliferation and differentiation of cervical squamous cell carcinoma SiHa cells and to investigate related signaling molecules. Methods: Primers were designed to amplify the full length of cyclin D1 gene and cyclin D1 gene was amplified by PCR for constructing pcDNA3.1 plasmid vector. The construct was then transfected into SiHa cells, and the cells with stable overexpression of cyclin D1 were established, cyclin D1 gene and protein expression were detected by RT-PCR and Western blot, respectively. Cell growth curve was documented by MTT assay. CK7, E-cadherin, vimentin, Snail gene and protein expression in transfected cells were detected by RT-PCR and Western blot. RT-PCR was used to detect the mRNA expression of proliferation and differentiation-related genes like CDK4, CDK2, p21, p27, cyclin E, Rb, E2F, E6/E7 and Ki-67. After synchronization of cells, RT-PCR was used to detect of cyclin D1 and p21 mRNA expression at different time points of the cell cycle. Results: The G-3 cells with cyclin D1 overexpression were successfully established. The growth curve and Ki-67 mRNA expression accelerated in G-3 cells.Vimentin and Snail expression significantly increased at both gene and protein levels, while E-cadherin, CK7 gene and protein expression significantly decreased, indicating epithelial mesenchymal transitionoccurred in G-3 cells.Meanwhile, mRNA expression of cyclin D1, CDK4, CDK2, p21, p27, cyclin E, E2F and Rb increased, while E6/E7 and p16 showed no significant change. The expression trends of p21 and cyclin D1 were almost identical with fluctuation at different time points in the cell cycle. Conclusions Overexpression of cyclin D1 induced by gene transfection promotes proliferation and epithelial mesenchymal transition in SiHa cells.The process is accompanied by up-regulation of CDK4, CDK2, p21, p27 and cyclin E genes.p21 expression increases synchronously with cyclin D1, suggesting a regulatory role in epithelial mesenchymal transition by affecting expression of vimentin in G-3 cells.

OBJECTIVETo investigate the expression of cyclin D1 in cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma and its relationship with human papillomavirus 16 (HPV16) E7 gene expression.

METHODSBoth SiHa and Hcc94 cell lines were obtained from cervical epithelial cells of squamous cell carcinoma. E6/E7 gene was silent in Hcc94 cell line.Expression levels of cyclin D1 mRNA and protein in CIN and squamous cell carcinoma were detected by QT-PCR and immunohistochemistry (IHC) respectively. SiRNA was constructed for targeting the promoter of HPV16 E7 and then transfected into SiHa cells to establish cm-16 line with stable silencing of E7. Control cell line B3 was obtained by blank plasmid transfection into SiHa cells. RT-PCR and Western blot were used to detect cyclin D1 mRNA and protein expression in the SiHa, B3, and cm-16 cells, respectively.

RESULTSCyclin D1 was expressed in the basal cells of normal cervical squamous epithelia and the expression gradually decreased in the progression from CIN1 to CIN3. Squamous cell carcinoma showed negative or scattered expression of cyclin D1 (P<0.05). Both mRNA and protein of cyclin D1 in E7(+) SiHa cells were lower than those in cm-16 and Hcc94 cells.

CONCLUSIONSquamous cell carcinoma with high HPV E7 expression shows low level of cyclin D1, suggesting that HPV16 E7 gene inhibits the expression of cyclin D1.

Carcinoma, Squamous Cell ; metabolism ; virology ; Cell Line, Tumor ; Cervical Intraepithelial Neoplasia ; metabolism ; virology ; Cyclin D1 ; genetics ; metabolism ; Female ; Human papillomavirus 16 ; Humans ; Immunohistochemistry ; Papillomavirus E7 Proteins ; genetics ; Promoter Regions, Genetic ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Uterine Cervical Neoplasms ; metabolism ; virology