Objective To investigate the effects and mechanisms of prostaglandin E2 receptor subtype 3 (EP3) on transforming growth factor β1 (TGF-β1)-induced mouse mesangial cells damage. Methods Primary mouse mesangial cells were separated and cultured. Three siRNAs were synthesized and transfected into mesangial cells for silencing EP3 by LipofectamineTM 2000 and the best one was chosen. MCs were grouped into: (1)control group; (2)TGF-β1 (10 μg/L) group; (3)NC-siRNA plus TGF-β1 (10 μg/L) group; (4) EP3-siRNA group; (5)EP3-siRNA plus TGF-β1 (10 μg/L). Then the proliferation of MCs was evaluated by CCK-8 assay. The expression of PGE2 and cAMP in cell supernatant were detected by ELISA. The mRNA and protein expression of fibronectin (FN), connective tissue growth factor (CTGF), cyclooxygenase-2 (COX2), membrane-bound prostaglandin E2 synthase 1 (mPGES1) were detected by real - time quantitative PCR and Western blotting. The phosphorylation of p38 MAPK and ERK1/2 was decected by Western blotting. Results Compared with control group, the cell proliferation induced by TGF-β1 was increased (P<0.05), the expression of PGE2 and cAMP were improved, mRNA and protein expression of FN, CTGF, COX2 and mPGES1 were up-regulated (all P<0.05). Compared with TGF-β1 group, the cell proliferation in EP3-siRNA plus TGF-β1 group was reduced, the expression of FN, CTGF, COX2 and mPGES1 mRNA and protein were downregulated (all P<0.05), the phosphorylation of ERK1/2, p38 MAPK were also declined (P<0.05). Conclusion EP3-siRNA may reduce TGF-β1-induced cell damage through upregulating the expression of cAMP, repressing the activity of ERK1/2 and p38 MAPK, inhibiting the expression of COX2 mPGES1 and PGE2 by feedback, then decreased the expression of FN and CTGF.

Background Fibrin glue has been utilized to adhere the graft during the pterygium excision with conjunctival autografts.Several relevant clinical randomized controlled trial(RCT) and retrospective studies have been published abroad,but the samples for its effectiveness and safety issue of fibrin glue and suture are still underinvestigation.Objective Current study was to quantificationally assess the efficacy and safety of fibrin glue versus sutures in the application of pterygium excision with conjunctival autografts.Methods Based on established search strategy,a computerized literature search was conducted to identify all citations concerning the RCT for effectiveness and safety evaluation of fibrin glue and suture for the graft fixation during the pterygium excision with conjunctival autografts from MEDLINE ( from 2000 to October,2010 ),EMbase ( from 2000 to October,2010 ),Cochrane Central Register of Controlled Trials ( Issue 4,2010 ),CBMdisc ( 2000 to October 2010 ),CNKI ( 2000 to October 2010 ),and the relevant conference proceedings and references searched by hand was performed as supplement.The included literature was scored with Jadad table.The Cochrane Collaboration' s RevMan 5.0 software was used for the test of heterogeneity or test for overall effect.The effective indexes,such as operative duration,recurrence rate and complication,were evaluated by Meta analysis.Results Six RCTs involving 401 eyes of 377 participants were identified.These literatures were published with English in 2004-2010 from China,New Zealand,Sweden,Israel,Turkey and Malasia and the Jadad scores were 4-5.The quantitatively analysis revealed that fibrin glue appeared to short the operative time compared with suturing method (MD =14.23 ;95％ CI:- 16.18- 12.29;P=0.00) and drop the rate of recurrence ( RR =0.49,95％ CI:0.26 -0.95 ; P =0.03 ).No significant differences were found in the rate of postoperative graft dehiscence or absence (RR =3.41,95 ％ CI:0.85-13.68;P =0.08 ).Conclusions Fibrin glue shows the good effectiveness and easy application during the pterygium excision with conjunctival autografts.Long-term follow-up of multi-central RCTs with a larger number of cases are still needed to support this conclusion.

Objective To evaluate the diagnostic value of ultrasound-guided pleural biopsy combined with thoracic biochemical detections in malignant and tuberculous pleural effusions. Methods Sixty-four patients with moderate or large pleural effusions and pleural thickening received the ultrasound-guided diagnostic pleural biopsy. All patients had chest CT enhancement scans to find out the suspicious pleural thickening preoperatively, facilitating the selection of puncture sites by ultrasound. Pleural tissue samples were sent for pathological examinations immediately. After successful achievements of pleural biopsy, ultrasound-guided aspiration or drainage was performed to alleviate symptoms, more importantly, to get pleural effusions for biochemical analysis. Biological results including carcinoembryonic antigen(CEA), CA125, CYFRA21 and lactate dehydrogenase(LDH) in malignant and tuberculous effusions were analyzed by group design t tests. The positive rates of CEA, CA125, CYFRA21, LDH in malignant and tuberculous effusions were compared by chi square tests. Results Pleural tissues in all cases were got by one pleural biopsy procedure. The strategy of pleural biopsy we used in this study had a successful rate reaching 100%(64/64), and 73% (46/64) patients had a definitive diagnosis as malignant or tuberculous effusion. Twenty-seven cases were diagnosed as malignant effusions and thirty-seven cases as tuberculous effusions based on the deifnitive clinical diagnosis. The positive rates of CEA, CA125, CYFRA21, LDH in malignant effusions were 100%(27/27), 100%(27/27), 100%(27/27), 89%(24/27) respectively, and 0%(0/37), 84%(31/37), 78%(29/37), 76%(28/37) respectively in tuberculous effusions. The positive rate of CEA between malignant and tuberculous effusions differed signiifcantly (χ2=64.0, P < 0.01), so did CA125 (χ2=3.1, P < 0.01) and CYFRA21(χ2=4.8, P<0.01). The average levels of CEA, CA125, CYFRA21, LDH in pleural effusion were (727.1±658.8)μg/L, (795.2±1249.6)×103 U/L, (296.2±320.7)μg/L, (1077.9±1058.5) U/L respectively, and (1.7±1.1)μg/L, (336.3±208.6)×103 U/L, (20.7±14.9)μg/L, (309.2±182.7) U/L in tuberculous effusions.There were signiifcant differences in CEA, CYFRA21 and LDH concentrations among malignant and tuberculous effusions (t=45.1, 27.4, 18.8 respectively, all P<0.01). Conclusion Ultrasound-guided pleural biopsy combined with CEA, CYFRA21 and LDH in pleural effusions had an important value in the etiological diagnosis of pleural effusions, while CA125 showed little value in the differential diagnosis.

OBJECTIVETo investigate the correlation of vasectomy with the risk of prostate cancer in Chinese men.

METHODSWe systematically searched the databases CNKI, VIP, Wanfang, PubMed, Embase, and Cochrane Library for the literature relating the relationship between vasectomy and the risk of prostate cancer in Chinese males up to December 2014. According to the inclusion and exclusion criteria, two investigators independently selected the eligible publications, evaluated their quality, and extracted relevant information, followed by a meta-analysis with the software STATA 12.0.

RESULTSNine studies were included in the analysis involving 1 202 cases of prostate cancer and 4,496 controls. Random-effect model analysis revealed no statistically significant correlation between vasectomy and the risk of prostate cancer (OR = 1.05; 95% CI 0.62-1.79), with an obvious heterogeneity (P < 0.001, I2 = 85.7%). No significant publication bias was found among the included studies (Egger, P = 0.824; Begg, P = 0.348).

CONCLUSIONThe results of our meta-analysis do not support the association of vasectomy with the increased risk of prostate cancer in Chinese population.

Asian Continental Ancestry Group ; China ; Humans ; Male ; Prostatic Neoplasms ; ethnology ; etiology ; Risk Assessment ; Vasectomy ; adverse effects

Objective To compare the curative effect of open and minimally invasive treatment of acute closed achilles tendon rupture with 5 years followed-up study.Methods From September,2010 to January,2012,28 patients with acute closed Achilles tendon rupture in our hospital were followed up for 5 years.There were 21 males and 7 females.Minimally invasive percutaneous suture in 11 cases;open suture in the treatment of 17 cases.The patients were followed up at 6 months,1 year,2 years,3 years,4 years,and 5 years after AOFAS and ATRS score.Results The follow-up time ranged from 60 to 72 months.All incisions healed by first intention and no incision related complications occurred.2 groups of patients with ATRS score at 6 months after operation:the open group was 81.23±3.99,minimally invasive percutaneous group of 88.27±4.27,the difference between the two groups was statistically significantly.After 1 year,there was no significant difference in the scores between the two groups.Two groups of patients with AOFAS score at 6 months after operation:the open group was 69.00±6.23,minimally invasive percutaneous group of 79.27±4.83,the difference between the two groups was statistically significantly.At one year after operation,the open group was 85.53±3.38,and the minimally invasive group was more than 89.90±3.38.The difference between the two groups was statistically significantly.After 2 years,there was no significant difference in the scores between the two groups.Conclusion There is no significant difference between the 2 years after surgery in the treatment of acute closed Achilles tendon rupture or open surger.

Objective To explore the application of hydrogen proton magnetic resonance spectroscopy (1H-MRS) in the diagnosis of peripheral tumor cell infiltration of gliomas. Methods Forty patients with glioma were examined by 1H-MRS preoperation, and were divided into low grade glioma group (n=20) and high grade glioma group (n=20) according to postoperative pathological diagnosis. Tumor resection with peripheral tissues marked previously was carried out under the guidance of neuronavigator system. All the pathological sections were divided into positive group and negative group according to the presence or absence of tumor cells, and the differences in pathological findings of peripheral regions (region 1, 2 and 3) and 1H-MRS values were analyzed in these two groups. Results No infiltration was found in the peripheral regions of low grade glioma group except for one case in peripheral region 1, while infiltration was found in all peripheral regions of high grade glioma group. There was no significant difference in 1H-MRS values between positive group (n=24) and negative group (n=36) in patients with high grade glioma (P>0.05). Conclusion 1H-MRS enjoys some advantages over routine radiological examinations in the diagnosis of peripheral tumor cell infiltration of gliomas. Total removal can be expected when combined with neuronavigator system, while there is room for improvement for relevant techniques.

Objective To generate hemogenic endothelial cells(HECs)from human induced pluripotent stem cells (hiPSCs)in vitro in order to learn more about the mechanism by which the vascular niche affects HECs production and self -renewal.Methods hiPSCs with reporter gene runx1c were differentiated to hematopoietic cells by spinEB method.The CD34 positive cells were sorted by magnetic-activated cell sorting(MACS)at day 10 after hematopoietic differentiation. Afterwards,these CD34 positive cells were co-cultured with DLL4 overexpressed vascular niche cells VeraVec to further differentiate to HECs.The HECs derived from the hiPSCs were characterized by FACS.Results We first established an hiPSCs single cell culture method for spinEB differentiation.Single cell cultured hiPSCs with reporter gene runx 1c were differentiated to form embryonic bodies(EBs)by spinEB method.The HECs were enriched from the day 10.Meanwhile, we cultured the E4ORF1 transfected human umbilical vein endothelial cell(HUVEC)line(VeraVec)and examined the expression of NOTCH signaling pathway related genes.According to the results, VeraVec had a high expression level of NOTCH ligand DLL4 at both mRNA and protein levels.And the CD34 positive HECs were co-cultured with DLL4 overexpressed VeraVec cells,which promoted the expression of tdTomato during hematopoitic differentiation and increased HSCs production.Conclusion A method of inducing hiPSCs differentiation by spinEB has been established, which can enrich HECs.This model can be applied to study the mechanism by which the vascular niche promotes hematopoietic differentiation from hPSCs.The generated functional HSCs are of great social and military values for HSCs transplantation and battlefield radiation injury treatment.

A new algorithm of adaptive super-resolution (SR) reconstruction based on the regularization parameter is proposed to reconstruct a high-resolution (HR) image from the low-resolution (LR) image sequence, which takes into full account the inaccurate estimates of motion error, point spread function (PSF) and the additive Gaussian noise in the LR image sequence. We established a novel nonlinear adaptive regularization function and analyzed experimentally its convexity to obtain the adaptive step size. This novel algorithm can effectively improve the spatial resolution of the image and the rate of convergence, which is verified by the experiment on optical images.
Algorithms ; Image Processing, Computer-Assisted ; methods ; Motion ; Time Factors

The Notch signaling pathway has been implicated in the regulation of cell-fate decisions such as differentiation of embryo stem cells and neural stem cells into neurons. We cultured human mesenchymal stem cells (hMSCs) in vitro and induced hMSCs to differentiate into neural cells by beta-mercaptoethanol (beta-ME), DMSO and 3-tert-butyl-4-hydroxyanisole (BHA). Immunocytochemistry was utilized to detect neuron-specific enolase (NSE) and Nissl body, and flow cytometry was used to determine cell growth phases. The expressions of signal molecules involved in the Notch pathway such as Notch1, Jagged 1 (JAG1), presenilin 1 (PS1) and hairy and enhancer of split 1(HES1) were observed by RT-PCR and immunofluorescent techniques. The results were as follows: (1) Before induction, the percentage of hMSCs at G(0)/G(1) was 58.5%, and the percentage at S+G(2)/M was 41.5%. After induction, the percentage of hMSCs at G(0)/G(1) increased to 73.1%, 76.2% and 78.1%, respectively on days 2, 4 and 6, and the percentage at S+G(2)/M decreased to 26.8%, 24.8% and 21.9%, respectively; The percentage of NSE-positive cells reached (77+/-0.35) %; Nisslos staining was positive in cytoplasm. (2) Notch1 and JAG1 were both expressed in hMSCs before and after induction, but the mRNA expressions of both Notch1 and JAG1, detected by RT-PCR, decreased obviously after induction(P<0.05). Notch1 mRNA/beta-actin was 1.157, 0.815, 0.756 and 0.570, and JAG1 mRNA/beta-actin was 0.437, 0.350, 0.314 and 0.362, respectively, on days 0, 2, 4 and 6 after induction. The Notch pathway activation participant PS1 mRNA and Notch pathway target gene HES1 mRNA also decreased apparently after induction (P<0.05), and their mRNA/beta-actin was 0.990, 0.449, 0.441, 0.454 and 0.370, 0.256, 0.266, 0.240 on days 0, 2, 4 and 6, respectively. These observations indicate that the expressions of Notch signal molecules were suppressed when hMSCs were induced to differentiate into neural cells. Based on these findings, we propose that low level of Notch signaling activation may contribute to neural cell differentiation.
Basic Helix-Loop-Helix Transcription Factors ; genetics ; Calcium-Binding Proteins ; genetics ; Cell Cycle ; Cell Differentiation ; Flow Cytometry ; Homeodomain Proteins ; genetics ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jagged-1 Protein ; Membrane Proteins ; genetics ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Receptor, Notch1 ; genetics ; Receptors, Notch ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Serrate-Jagged Proteins ; Signal Transduction ; Transcription Factor HES-1

OBJECTIVETo detect the immunoreactivity of targeted fusion anti-caries DNA vaccine pGJA-P in vitro, and the ability to enhance the immune responses compared with the non-targeted fusion anti-caries DNA vaccine pGLUA-P.

METHODSThe CHO cells were transfected with pGJA-P and the expression of recombinant protein in cultured supernatants were detected using Western blotting. 5 to 6-month-old female Japanese rabbits were immunized with either pGJA-P or pGLUA-P via either intramuscular injection (i.m.) or intranasal route (i.n.). The sera and saliva were collected and the antibody responses were checked by ELISA. The effect of immune sera on the synthesis of water-insoluble glucan by glucosyltransferase of S. mutans was examined.

RESULTSThe expressed protein could response to specific anti-GTF antibody. The antibody responses in serum generated by pGJA-P via i.m. were significantly higher than those generated by pGLUA-P (P < 0.01). The antibody responses in saliva generated by pGJA-P via i.n. were significantly higher than those generated by pGLUA-P (P < 0.01). The higher mucosal antibody response induced by pGJA-P via i.m. compared with pGLUA-P (P < 0.01) was detected. The immune sera of rabbits immunized by pGJA-P via i.m. most significantly inhibited the synthesis of water-insoluble glucan by glucosyltransferase.

CONCLUSIONSThe recombinant protein expressed by pGJA-P had the immunoreactivity to anti-GTF antibody. pGJA-P could induce faster and higher specific mucosal SIgA antibody responses via i.n. or serum IgG antibody responses via i.m. compared with non-targeted DNA vaccine, pGLUA-P. High titres of specific mucosal antibodies were found in rabbits immunized with pGJA-P via i.m. The immune sera of rabbits immunized by pGJA-P via i.m. displayed the ability of inhibiting the synthesis of water-insoluble glucan by glucosyltransferase.

Animals ; Bacterial Proteins ; immunology ; CHO Cells ; Cricetinae ; Dental Caries ; prevention & control ; Female ; Glucosyltransferases ; immunology ; metabolism ; Immunoglobulin G ; blood ; Membrane Glycoproteins ; immunology ; Rabbits ; Recombinant Fusion Proteins ; administration & dosage ; immunology ; Streptococcus mutans ; immunology ; Transfection ; Vaccines, DNA ; administration & dosage ; immunology