Objective To investigate the risk factors associated with neovascular glaucoma (NVG) after pars plana vitrectomy (PPV) in eyes with proliferative diabetic retinopathy (PDR).Methods Retrospective study.One hundred and thirty-seven patients (137 eyes) with PDR who underwent PPV were recruited.There were 85 males and 52 females.The average age was (60.1 ± 8.8) years old.The duration of diabetes was (10.2 ± 3.6) years.There were 49 patients with ipsilateral carotid artery stenosis.Fifty-three eyes underwent intravitreal ranibizumab or conbercept injection before PPV.All eyes were treated with 23G standard three-port PPV.The average follow-up time after PPV was 11.5 months.Fundus fluorescein angiography (FFA) was conducted in postoperative 4-6 weeks to observe non-perfused retinal areas.Risk factors,such as ipsilateral carotid artery stenosis,the presence of non-perfusion in retina after PPV and the application of anti-vascular endothelial growth factor (VEGF) drugs before PPV,were identified by logistic regression.Results Twenty of 137 patients (14.6％) developed postoperative NVG after PPV.Ipsilateral carotid artery stenosis [odds ratio (OR) =5.048,95％ confidence interval (CI) 2.057-12.389,P=0.000] and the presence of non-perfusion in retina after PPV (OR=4.274,95％CI 1.426-12.809,P=0.009) were significant risk factors for postoperative NVG,while the application of anti-VEGF drugs was not (OR=1.426,95％CI 0.463-4.395,P=0.536).But the time from PPV to the onset of NVG varies significantly between the two groups of injection of anti-VEGF drugs or not (t=-4.370,P=0.000).Conclusions Risk factors associated with NVG after PPV in eyes with PDR included ipsilateral carotid artery stenosis and the presence of non-perfusion in retina after PPV.The application of anti-VEGF drugs before PPV can delay the onset of NVG in PDR eyes after vitrectomy.

Objective To analyze the expression feature and function of microRNAs in exosomes secreted by leukemia cells (LCEX). Methods The mice leukemia cell line L1210 was taken as the example, and LCEXL1210 was obtained by isolating supernate of L1210 cells through density gradient centrifugation. MicroRNAs isolated from LCEXL1210 were analyzed by microarray analysis, compared with miRNA from L1210 cell line, and then some of miRNAs with different expression were verified by real-time PCR and were analyzed by Gene Ontology (GO) database. Results The number of miRNAs identified in LCEXL1210 was 1 044, and that in L1210 cell line was 872. The number of shared miRNAs between LCEXL1210 and L1210 cell line was 732, accounting for 70.1 % of LCEXL1210 and 83.9 % of L1210 cell line, respectively, which indicated that 70 % of LCEXL1210 was derived from the parental cells. Interestingly, 312 miRNAs in LCEXL1210 were found to be underrepresented in the parental cells, indicating their specificity in LCEXL1210. Some miRNAs were significantly highly expressed in LCEXL1210 compared with those in L1210 cell line, including miR-16-1, miR-210, miR-195 and so on, which showed that miRNAs isolated from LCEXL1210 were differentially expressed with those from the parental cells. Some differentially expressed miRNAs from LCEXL1210 were verified by real-time PCR, and then were analyzed by GO database, which demonstrated that these highly expressed miRNAs participated in the processes of various biological function and signal transduction. Conclusions MiRNAs isolated from LCEXL1210 show a high similarity to miRNAs isolated from L1210 cells, whereas of which one-third are specific. The highly expressed miRNAs participate in the processes of various biological function and signal transduction.

ObjectiveTo observe the effect of different extracts ofTangwang Mingmu Granule on hypoxia induced gene expressions in vascular endothelial cells.Methods COCl2 intervention cells were used to copy hypoxia models. Human umbilical vein endothelial cells EA. Hy926 were divided into blank group, hypoxia model group,Tangwang Mingmu Granule group, extract 1 (glycosides and flavonoids) group, extract 2 (orgain acids and polysaccharides) group and extract 3 (alkaloids) group. The changes in gene expressions of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α mRNA were detected by semi-quantitative RT-PCR.ResultsThe gene expression levels of VEGF, VEGFR-1, VEGFR-2, ICAM-1 and IL-1α were significantly up-regulated under the hypoxic condition (P<0.05), and the ratio of VEGFR-1/VEGFR-2 was significantly reduced. However,Tangwang Mingmu Granule significantly reversed the expressions of these genes.Conclusion The function intensity of gene expressions weakens in the following sequence:glycosides and flavonoids>alkaloids>organic acids and polysaccharides inTangwang Mingmu Granule.

Objective To observe the mechanism of action of different extracts ofTangwang Mingmu Granules on high glucose induced VEGF and IL-1α gene and protein expressions in vascular endothelial cells.Methods Human umbilical vein endothelial cells EA.hy926 were divided into six groups: blank, high glucose,Tangwang Mingmu Granules, extract 1 (glycoside and flavonoid), extract 2 (organic acid and polysaccharides) and extract 3 (alkaloids) groups. High concentration glucose was used to establish the high glucose model in EA.hy926 cells. The expressions of VEGF and IL-1α mRNA were detected by semi-quantitative RT-PCR. The contents of VEGF and IL-1α protein were tested by ELISA.Results The gene expression and protein levels of VEGF and IL-1α were significantly up-regulated under the high glucose condition (P<0.05). However, the above indicators were significantly reduced after the treatment ofTangwang Mingmu Granules. The activity of different parts ofTangwang Mingmu Granules was as follows: extract 1> extract 3> extract 2.Conclusion The action intensity of glycosides and flavonoids, alkaloids, organic acids and polysaccharides on VEGF and IL-1α expression inTangwang Mingmu Granules weakens in sequence.

Objective To observe vasculogenic mimicry formation difference in human hepatocellular carcinoma cell lines MHCC97-H/L with different metastatic potentials under three-dimensional culture condition,and to study extracellular matrix and adhesion molecules-related mechanism of vasculogenic mimicry.Methods Three-dimensional cell culture system of MHCC97-H/L was established to observe tubelike structures by inverted microscope.Human umbilical vein endothelial cell(HUVEC),human hepatocellular carcinoma cell line Hep3B with non-metastatic potentials and normal human hepatic cell line HL-7702 were taken as control.Gene expression profiles of MHCC97-H and MHCC97-L were detected by Oligo extracellular matrix and adhesion molecules microarray analysis.Two differentially expressed genes were verified with semi-quantitative reverse transcriptionpolymerase chain reaction(RT-PCR)and Western blot.All data were analyzed using two sample t test.Results After a 24-hour three-dimensional culture,the length of tubelike structures was(474 ± 16)mm/cm2 in MHCC97-H,which were significantly longer than(320 ±41)mm/cm2 in MHCC97-L(t =6.119,P ＜0.05).Hep3B and HL-7702 could not form tubelike structures.Compared with MHCC97-L,the expression of 7 genes were up-regulated and the expression of 3 genes were down-regulated among a total of 113 angiogenesis genes in MHCC97-H.Two of the genes with differential expressions including tenascin C and extracellular matrix protein 1 were further validated by RT-PCR and Western blot.Conclusion MHCC97-H has stronger ability to form vasculogenic mimicry than MHCC97-L,and this perhaps correlates with the differential expression of certain extracellular matrix and adhesion molecules-related genes in MHCC97-H.

In this article, the status of spindle assembly checkpoint and the alteration of its major component, Mad2 protein level were examined in A2780 and SKOV3 ovarian cancer cell lines. Recombinant eukaryotic expression plasmid pEGFP-Mad2 was transfected into paclitaxel-resistant SKOV3 cells and Mad2 protein was knocked down by Mad2-specific siRNA in paclitaxel-sensitive A2780 cells. Then the expression level of Mad2 gene was detected by Western blotting. Flow cytometry revealed that SKOV3 cells were not fully arrested in G(2)/M phase in contrast to A2780 cells in the presence of paclitaxel. However, paclitaxel sensitivity assay showed that sensitivity to paclitaxel was reversed after the transfection in both cell lines in terms of number of cells arrested at G(2)/M phase and the expression of Bcl-2 was significantly changed. These results suggest that weakened spindle checkpoint with reduced expression of Mad2 is associated with resistance to paclitaxel in ovarian cells and Bcl-2 may be involved in this process.

Aged ; Aged, 80 and over ; Humans ; Laparoscopy ; methods ; Male ; Prostate ; surgery ; Prostatectomy ; methods ; Prostatic Neoplasms ; surgery

Objective To induce human umbilical cord mesenchymal stem cells (hUC-MSCs) to hair-cell like cells in the inner ear, using a two-step neural differentiation method.Methods The hUC-MSCs were obtained from human umbilical cords by tissue adherence culture,whose surface antigen CD29, CD34, CD44, CD45, CD90, HLA-ABC, and HLA-DR could be identified by flow cytometry.In the neural stem cells induced phase, the NSE positive cells were analyzed by microscope and immunohistochemistry.In the second stage, the expression of hair-cell like cells markers (Math1, MyosinⅦa, Brn3c) were tested by qRT-PCR and immunofluorescence method.Results The control group and the protocol group had little NSE after differentiation while the protocol B group presented a neurobiological structure and demonstrated a higher NSE positive ratio after 5 days' neural stem cells induction (P<0.05).Compared to the control group, the mRNA and protein level of Math1, MyosinⅦa, and Brn3c exhibited a significant increase in the differential group,which induced for 4 weeks in the hair-cell like cells in the inner ear's induced phase(P<0.05).Conclusion The two-stage induction (hUC-MSCs-neural stem cells-hair-cell like cells) could produce more MyosinⅦa,Brn3c and Math1,which may provide an appropriate way to treat sensorineural deafness.

OBJECTIVESTo evaluate the application of two-dimensional electrophoresis and mass spectrometry in the research of proteins expressed in testis.

METHODSProtein from adult ICR mouse testes was extracted by two-dimensional electrophoresis. Two protein spots were cut from the gel, then the proteins were digested in-gel by enzyme and the generated peptides were measured by matrix assisted laser desorption/ionization-time of flight mass spectrometry. The proteins were identified through database searching.

RESULTSTwo spots in Commasie Brilliant Blue-stained gel were identified as serum albumin and protein disulfide isomerase by database searching.

CONCLUSIONSThis rapid high resolution and efficient method is a very powerful way to analyze testicular proteins.

Animals ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred ICR ; Proteins ; analysis ; Testis ; chemistry

PURPOSE: Triple-negative breast carcinoma (TNBC) is accompanied with high risk of metastasis and recurrence. The present study aimed to explore the clinicopathological and prognostic roles of putative tumor-related genes in patients with TNBC. METHODS: Thirty pairs of frozen-thawed tumors were used to select reliable indicators via real-time quantitative polymerase chain reaction (RT-qPCR). Then, 150 pathology specimens were used to evaluate the expression of proteins in TNBC through immunohistochemistry. In addition, Kaplan-Meier curves and Cox regression analysis were also performed to analyze the overall survival and disease-free survival. RESULTS: RT-qPCR results indicated that among all the proteins analyzed using fresh-frozen TNBC samples, the expression levels of only Survivin and zinc finger of the cerebellum 1 (ZIC1) were obviously different from those in the corresponding normal tissues. Survivin and ZIC1 expression had opposite effects on the clinicopathological diagnosis and prognostic assessment in TNBC patients. Further, there was a negative correlation between Survivin and ZIC1 expression. In addition, the “Survivin-positive ZIC1-negative group” was associated with histologic grade, lymph node metastasis, and TNM staging (p < 0.001) and this was also an independent factor for evaluating the prognosis of TNBC in patients. CONCLUSION: In summary, the expression levels of Survivin and ZIC1 in TNBC are different from those in normal tissues and are negatively correlated mutually. The combined detection of Survivin and ZIC1 expression levels could allow better comprehensive diagnosis and prognostic evaluation for TNBC patients.
Breast Neoplasms ; Breast ; Cerebellum ; Diagnosis ; Disease-Free Survival ; Humans ; Immunohistochemistry ; Lymph Nodes ; Neoplasm Metastasis ; Neoplasm Staging ; Pathology ; Polymerase Chain Reaction ; Prognosis ; Recurrence ; Triple Negative Breast Neoplasms ; Zinc Fingers ; Zinc