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The effects of highly expression of polypyramidine tract binding protein-associated splicing factor on advanced glycation end-products-induced human retinal microvascular endothelial cells

ObjectiveTo investigate the protection and the corresponding molecular mechanisms of polypyramidine tract binding protein-associated splicing factor (PSF) overexpression on human retinal microvascular endothelial cells (hRMECs) induced by advanced glycation end-products (AGEs).MethodsThe hRMECs were divided into the normal group, the vector group, PSF group, zinc protoporphyrin (ZnPP) group and PSF+ZnPP group for experiment. Cells in the normal group were cultured in a DMEM medium containing 10% fetal calf serum, penicillin/streptomycin, and placed in a closed constant temperature incubator at 37 °C, 95% air, and 5% CO2. Cells in the vector group were infected with empty lentivirus. The cells in the PSF group were infected with overexpressing PSF lentivirus. Cells in the ZnPP group were treated with ZnPP (10 mol/L) for 2 h. The PSF+ZnPP group cells were infected with overexpressing PSF lentivirus, and then pretreated with ZnPP (10 mol/L) for 2 h. With the last four groups of cells stimulated with AGEs, HE, Hoechst33258 staining and flow cytometry were used to observe the protective effect of high expression of PSF on cell damage and the antagonistic effect of ZnPP on PSF. Western blot was used to detect the protein expression of heme oxygenase-1 (HO-1), phosphorylated (p) extracellular regulatory protein kinase (ERK), and Nrf2 in the cells. U0126, a specific antagonist of ERK pathway, was introduced, and Western blot verified the reversal effect of U0126 on the expression of HO-1 induced by PSF protein.ResultsHE staining and Hoechst33258 staining showed that the number of nuclei of damaged cells of PSF group were significantly increased compared with control group, while decreased compared with PSF+ZnPP group (F=27.5, 38.7; P＜0.05). The results of flow cytometry showed that the ROS produced by cells in the PSF group was significantly increased compared to the normal group, and significantly decreased compared to the PSF+ZnPP group, the difference was statistically significant (F=126.4, P＜0.05). Western blot results showed that HO-1 expression of PSF group was significantly increased compared with control and the vector group (F=70.1, P＜0.05). AGEs inducement of 30, 60, 120 and 240 min could significantly improve pERK expression compared with 15 min (F=474.0, P＜0.05). The expression of HO-1 and Nrf2 proteins in the PSF+/U0126－ group was significantly more than those in the PSF－/U0126－ group, the expression of HO-1 and Nrf2 proteins in the PSF+/U0126+ group was significantly lower than that in the PSF+/U0126－ group, and the differences were statistically significant (F=30.2, 489.4; P＜0.05).ConclusionOver expression of PSF can promote the HO-1 expression by activating ERK pathway and promoting the Nrf2 to the nucleus, thus protect hRMECs against AGEs-induced oxidative damage.

23389例西部地区健康女性宫颈糜烂情况及其相关因素分析

目的了解西部地区健康女性宫颈糜烂患病情况、年龄分布特点及其与宫颈细胞学之间的关系。 方法回顾性分析2011年12月－2012年11月体检女性的宫颈糜烂情况、宫颈细胞学的结果。 结果共纳入研究对象23 389例，宫颈糜烂总患病率为20.98%，18～30岁年龄段患病率最高，约有44.81%的女性发生宫颈糜烂，其中，31.62%的女性为轻度糜烂，9.83%的女性为中度糜烂，3.36%的女性为重度糜烂。宫颈糜烂的患病率随着年龄的增加呈递减趋势，该趋势有统计学意义（χ2=2310.238，P＜0.001）；剔除阴道炎的女性后，纳入细胞学分析的女性为22 477例，宫颈细胞学异常者194例，宫颈细胞学异常率为0.86%，其中无糜烂者宫颈细胞学异常率为0.59%，轻度糜烂者宫颈细胞学异常率为1.47%，中度糜烂者宫颈细胞学异常率为2.96%，重度糜烂者宫颈细胞学异常率为6.84%，随着宫颈糜烂的发生及程度的增加，宫颈细胞学异常率增加，趋势有统计学意义（χ2=127.472，P＜0.001）。 结论宫颈糜烂的情况非常常见，18～30岁年龄段女性患病率最高，随着宫颈糜烂的发生及程度的增加，宫颈细胞学异常率增加，然而，并不是宫颈糜烂之后就会出现宫颈上皮内瘤变或癌变，也并不是所有的宫颈上皮内瘤变的患者均有宫颈糜烂，阴道脱落细胞学的检测对于有性生活的女性是必不可少的体检项目。

The Research of Apoptosis in Conjunctiva Epithelial Cells with Connective Tissue Disease

Objective　To investigate pathogenesis of dry eye and applied value in diagnosis of dry eye with connective tissue disease(CTD) by apoptosis detection, using impression cytology flow cytometry (ICFC) in conjunctiva epithelial cells. Methods　A total of 60 patients (120 eyes) with CTD, after asked case history and measured the basal Schirmer’s test (S-I-T), Break-Up Time (BUT), fluorescent Staining (FL), were divided into 4 groups: the first group without Sjögren syndrome or dry eye (NSS1), the second group without Sjögren syndrome but dry eye (NSS2), the third group with Sjögren syndrome and non-dry-eye (SS1) and the fourth group with Sjögren syndrome and dry eye (SS2). And apoptosis of conjunctiva epithelial cells was detected by ICFC. Results　The apoptosis rate of conjunctiva epithelial cells was statistically significant (Plt;0.001) between every two groups, except that between NSS1 group and SS1 group (P=0.998). And apoptosis rate was a positive correlation with FL (r=0.926, Plt;0.001), but negatively with S-I-T and BUT (r= –0.712, r= –0.818, Plt;0.001). Dye eye and Sjögren-syndrome both affected the apoptosis level of conjunctiva epithelial cell and there was an interaction between them. Conclusion　Apoptosis plays an important role of ocular damage and apoptosis detection helps with diagnosis of dry eye with CTD. Dye eye and Sjögren-syndrome increase apoptosis level. Apoptosis detection by ICFC in conjunctiva epithelial cells is a minimally invasive and effective way to detect ocular apoptosis.

RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor treatment

ObjectiveTo observe RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.MethodsCultured the retinal vascular endothelial cells in vitro and logarithmic growth phase cells were used for experiments. The cells were divided into VEGF group and VEGF combined with anti-VEGF drugs group. The VEGF group cells were treated with 50 ng/ml VEGF for 72 h to simulate the high VEGF survival conditions of vascular endothelial cells in diabetic retinopathy. VEGF combined with anti-VEGF drug group cells was treated with 50 ng/ml VEGF and 2.5 μg/ml anti-VEGF drugs for 72 h to imitate the microenvironment of cells following the anti-VEGF drugs treatment, and whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq. Now with biological big data obtained as a basis, to analyze the differentially expressed genes (DEGs). And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.ResultsThe gene expression profiles of the two groups of cells were obtained. Through analysis, 328 DEGs were found, including 194 upregulated and 133 downregulated ones. The functions of DEGs were influenced by regulations over molecular biological process, cellular energy metabolism and protein synthesis, etc. Among these genes, SI,PRX and HPGD were related to protein synthesis, BIRCT to cellular apoptosis, and ABLIM1 and CRB2 to retinal development, and ABCG1, ABCA9 and ABCA12 were associated with the cholesterol of macrophage and the transfer of phospholipid. GO enrichment analysis showed that DEGs mainly act in three ways: regulating biological behavior, organizing cellular component and performing molecular function. Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in ECM receptor pathway, and Notch, mitogen-activated protein kinase, transforming growth factor (TGF)-β and Wnt signal pathways. Among them, the gene expression in TGF-β signal pathway attracts most attention, where the DEGs, such as CAMK2B, COL3A1, CYGB, PTGER2 and HS6ST2, among others, were closely related to fibrosis process.ConclusionThe anti-VEGF drugs may enhance the expression of CAMK2B, COL3A1, CYGB, PTGER2 and others genes related to TGF-β signal pathway and aggravate retinal fibrosis disease.

牛视网膜微血管内皮细胞的培养方法学原理和生长性状初探

Effect of subretinal injection of retinal pigment epithelial cells for retinal pigment epithelium in mice

ObjectiveTo observe the effect of subretinal injection of retinal pigment epithelium (RPE) cells for RPE in mice. MethodsA total of 30 postnatal day 7 C57BL/6J mice were randomly divided into normal mice group, OIR model group and OIR model cell transplanted group, 10 mice in each group. The OIR model was induced in mice of OIR model group and OIR model cell transplanted group. The RPE cells were subretinal injected into the RPE of mice in OIR model cell transplanted group. At 20 days after the injection, the RPE thickness was evaluated by fluorescence microscope. The expression of RPE65, Bestrophin and zonula occludens-1 (ZO-1) were estimated by Western blot and real-time quantitative PCR (RT-PCR). ResultsThe thickness of RPE in OIR model mice was thinner than that in normal mice; the thickness of RPE in OIR model cell transplantation mice was significantly thicker than that in the OIR model mice. The results of Western blot and RT-PCR indicated that the differences of protein (F=8.597, 18.864, 25.691) and mRNA expression (F=39.458, 11.461, 34.796) of RPE65, Bestrophin, ZO-1 were statistically significant between OIR model group and OIR model cell transplanted group (P < 0.05). ConclusionsSubretinal injection of RPE cells can promote RPE thickening. RPE65 and Bestrophin protein relative expression levels increased, ZO-1 protein relative expression levels reduced; mRNA expression levels of RPE65, Bestrophin and ZO-1 genes increased.

Clinical Analysis of 13 Patients with Cervical Cancer Associated with Pregnancy

目的:探讨妊娠相关性宫颈癌的早期诊断、治疗和预后。方法:结合文献回顾分析我院2000年至2007年收治的13例妊娠相关性宫颈癌的诊治经过和预后。结果:妊娠相关性宫颈癌分化程度低，癌灶体积大，早期盆腔淋巴结转移率高，产褥期宫颈癌预后差。结论:宫颈细胞学检查应列为首次产检常规项目；妊娠期宫颈原位癌在密切随诊前提下可暂不予处理，待分娩后6~8周活检确认病变性质后，再采取相应治疗措施；新辅助化疗同样可为晚期别的妊娠相关性宫颈癌争取手术时机。

Analysis of Cause of High Incidence of Cervical Cancer of Xinjiang Area from 2000 to 2005

摘要:目的: 调查新疆地区维吾尔族与汉族子宫颈癌及癌前病变发病情况,分析宫颈癌高发原因。 方法 : 2000年1月至2005年12月新疆自治区人民医院妇产科门诊及病房行宫颈细胞学检查的维吾尔族、汉族妇女进行筛查,对宫颈病变阳性者(CINI以上)行病理组织学检查,对结果进行对比分析、综合评价。 结果 : 宫颈涂片人数共计23 205例,其中维吾尔族6 999例、汉族16 206例。宫颈病变阳性者237例,经阴道镜下病理活组织检查证实CINI以上(包括CINI、CINII、CINIII、原位癌、鳞癌、腺癌)病变人数173例,最小年龄31岁,原位癌(维吾尔族)、最大年龄76岁,宫颈磷癌(汉族)。维吾尔族105例(6069%)、汉族68例(3931%)。每年阳性例数中维吾尔族均高于汉族,其中2000年、2001年、2004年、2005年有极显著性差异(P lt;001),2002年、2003年有显著性差异(P lt;005),维吾尔族、汉族在各年龄组中的发病情况无显著性差异(P gt;005)。 结论 : 新疆地区宫颈癌及癌前病变的高发原因是由地区环境、医疗条件、医学发展、救助措施等因素综合作用的结果。Abstract: Objective: To investigate the incidence of cervical cancer and cervical precancerous lesion of uigur nationality and han nationality, in addition, to analysis the cause of cervical cancer’s high incidence. Methods : At first screen cervical cytology of Uigur and Han outpatient and inpatient of department of gynecology and obstetrics in the People’s Hospital of Xinjiang Uigur Autonomous Region from January 1, 2000 to December 31, 2004Secondly biopsy for those patients that cervix pathological change shows positive, then contrast analysis and comprehensive evaluation. Results : Cervix smears are 23205 samples. Uigur nation has 6999 samples and Han nation has 16206 samples. There are 237 patients whose cervix pathological changes shows positive. Among them 173 samples were over CINⅠ(include CINⅠ,CINⅡ,CIN Ⅲ,carcinoma in situ, squamous carcinoma and adenocarcinoma) through colposcopy. The youngest was 31 and diagnosed carcinoma in situ(Uigur), the eldest was 76 and diagnosed squamous carcinoma(Han).The samples of Uigur is 105(6069%) and Han is 68(3931%).The positive samples in Uigur is higher than Han each year, the incidence has extremely significant difference among 2000,2001 and 2004(P lt;001), while it has significant difference between 2002 and 2003 (P lt;005), but in each age group it has no significant difference between Uigur and Han (P gt;005). Conclusion : The high incidence of cervical cancer and cervical precancerous lesion in xinjiang is contribute to environment, medical condition, medical development and aid measures coaffect.

Evidence-Based Lung Cancer Screening in the Early Stage

Lung cancer is the leading cause of death among the tumors in the whole world. Although new diagnostic techniques have been developed for nearly 20 years, the mortality is still high. Until now, no randomized controlled trial of chest x-ray and sputum cytology showed the improvement of the survival rate of lung cancer. Low-dose CT can screen more patients in early stage, however, overdiagnosis, cost and the quality of studies should be considered. Further studies of RCTs should be done to clarify these questions.

The Value of High Risk Human Papillomavirus Detection for Cervical Cancer Screening

【摘要】 目的 评价人乳头状瘤病毒(HPV)DNA检测在宫颈癌筛查中的价值。 方法 采用第二代杂交捕获(HCⅡ)技术和液基细胞学测试(LCT)2种方法,对1026例在妇科病中心就诊的受检者进行同步盲法检测,同时进行阴道镜检查。以宫颈活检组织病理学检查结果为诊断标准。评价该方案在宫颈癌筛查中的应用价值。 结果 病理检查结果显示,宫颈上皮内瘤变(CIN)Ⅰ级152例,CINⅡ级108例,CINⅢ级109例,宫颈浸润癌28例。筛查高危型HPV感染366例,阳性率3570%, 在不同宫颈病变中的阳性率分别是:宫颈癌9290%(26/28),CINⅢ900%(99/109),CINⅡ8890%(96/108),CINⅠ8750%(133/152)。高危HPV对宫颈高级别病变的敏感性、特异性、阳性预测值,阴性预测值分别是9860%、8610%、1480%和9980%;HPV与LCT联合检测(平行试验)的以上各指标分别是10000%、8090%、1210%和10000%。 结论 高危型人乳头状瘤病毒检测在宫颈癌前病变的筛查中有较高的敏感度和阴性预测值,联合LCT检测是目前宫颈癌筛查具有诊断价值的方法。【Abstract】 Objective To investigate the value of high risk human papillomavirus(HPV) DNA dectection for cervical cancer screening. Methods Hybrid capture Ⅱ(HCⅡ)human papillomavirus (HPV) test and liquid based cytology test (LCT) were performed in 1026 patients treaed in Xuzhou No.1 hospital from May 2008 to May 2009,and the abnomal cytological or HPV DNA findings were further biopsied under the colposcopeto to appraise the appicational importance of each approach for screening cervical cancer. Results Pathological results showed that cervical intraepithelial neoplasial(CIN)Ⅰin 152 patients,CIN Ⅱ in 108 patients,CIN Ⅲ 109 patients,invasive cervical cancer in 28 patients.HPV infected 366 patients in detection, with 3570% positive rate. The infection rate of HPV in cervical cancer was 929%(26/28),in CIN Ⅲ was 908%(99/109),in CIN Ⅱ was 889%(96/108),and in CIN Ⅰwas 875%(133/152).The pathological results treated as standard,the sensitivity, soecificiy, positive prevalue, negative prevalue of HCⅡ HPV for detecting highgrade cervical lesions were 986%,861%,148% and 998%.The values for HPVLCT parallel test were 1000%,809%,121% and 100%. Conclusion Highrisk HPV DNA test is of high sensitivity and negativepredictive value. The combination of HCⅡ HPV and LCT tests are of great value for screening cervical cancer at present.