Abstract:Objective:To investigate the effect of desmin on embryo development of Xenopus laevis. Methods:The expression of desmin during embryogenesis was detected by real-time RT-PCR and the whole mount in situ hybridization. The specific antisense oligonucleotides was performed to knockdown the expression of desmin by microinjection. The expression of marker genes was tested during germ layer formation by whole mount in situ hybridization and quantitative RT-PCR. Results:The weak expression of desmin starts from the early blastula stage and then detected through all periods of organogenesis. Knockdown of desmin inhibited the blastopore closure,and suppressed the expression of the endodermal,mesodermal and exodermal marker genes. Conclusion:The expression of desmin starts from the blastula during embryogenesis of Xenopus lavies. Loss function of desmin leads to embryonic growth retardation and depressed germ-layer formation.

Abstract:Objective:We sought to culture and isolate cochlear progenitor cells (CPCs),identify their proliferation and differentiation capacity,and demonstrate the expression of genes involved with the development of inner ear during the CPCs differentiation. Methods:CPCs were isolated from newborn rats’ cochlea,in vitro cultured and differentiated by serum. Proliferation and differentiation of hair cells were identified by immunocytochemistry method. The expressions of Sox2,CyclinA2,and Jagged1 genes during CPCs differentiation were analyzed using real time qRT-PCR. Results:The cultured CPCs expressed nestin and BrdU,indicating the proliferation ability of CPCs undergoing mitosis. The expression of myosin Ⅶ A in the CPCs-derived differentiated cells indicated the CPCs’ potential to differentiate into hair cells. It was also noteworthy that the expressions of Sox2 and CyclinA2 were downregulated and the Jagged1 expression was upregulated during the CPCs differentiation. Conclusion:The CPCs in vitro culture showed that it has the ability of proliferation and potential of differentiation into hair cells. Additionally,it was suggested that Sox2,CyclinA2 and Jagged1 genes could potentially participate in the modulation of CPCs differentiation and development of cochlear tissues in rats.

Abstract:Objective:To detect the expression of IL-2 receptor β and γ chain(CD122 and CD132) on CD4+CD25-T cells in peripheral blood of systemic tupus erythematosus (SLE)patients and CD4+CD25+T cells which were induced from the former in vitro,as well as assess its effects on the induction of Foxp3+ induced regulatory T cell (iTreg). Methods:Peripheral blood mononuclear cells (PBMCs) were isolated from SLE patients and healthy donors and further isolated CD4+CD25-T cells. The ratio of CD25-CD122+/CD4+,CD25-CD132+/CD4+ and CD122/CD132 (IL-2 receptor β and γ chain) mRNA relative abundance in CD4+CD25-T cells were analyzed by flow cytometry and real-time quantitative PCR,respectively. CD4+CD25-T cells were induced to further analyze the ratio of CD25+CD122+/CD4+,CD25+CD132+/CD4+,CD25+Foxp3+/CD4+,pStat5+/CD4+CD25+. Results:Compared with the normal subjects,there were no significant changes in the expression of CD122 on CD4+CD25-T cells in SLE patients PBMCs,but the ratio of CD25-CD132+/CD4+ was lower and negatively correlated with the SLEDAI;The CD132 mRNA relative abundance in CD4+CD25-T cells was also lower than that in the control group;After CD4+CD25-T cells in peripheral blood of SLE patients were induced,the percentage of CD25+Foxp3+T cells in CD4+T cells was lower than that in the control group and negatively correlated with the SLEDAI;The ratio of CD25+CD122+/CD4+and CD25+CD132+/CD4+ in cells which were transformed from CD4+CD25-T cells in SLE patients was lower than that in the control group,but only the latter was negatively correlated with SLEDAI. The expression of phosphorylated Stat5 in transformed cells of SLE patients was lower than that in the control group and the ratio of pStat5+/CD25+CD4+ was negatively correlated with SLEDAI and positively correlated with the ratio of CD25+Foxp3+/CD4+. Conclusion:There was obviously defective expression of CD132 (IL-2Rγ chain) on CD4+CD25-T cells in peripheral blood of SLE patients and it was correlated with disease activity;The expression of CD132 and CD122 (IL-2Rβ chain) was also defective when the cells were transformed into CD4+CD25+T cell subsets,but only the defective expression of CD132 was correlated with disease activity and Foxp3 expression and was accompanied by lower levels of phosphorylated Stat5,suggesting that the defective expression of IL-2Rγ chain and its weaker downstream signal are closely related with SLE patients Foxp3+iTreg induction obstacle.

Abstract:Objective:To investigate the correlation between E-cadherin and miR-148a,and explore the underlying mechanism of miR-148a regulating E-cadherin expression in gastric cancer. Methods:The mRNA levels of E-cadherin and miR-148a were detected by qRT-PCR,and protein expression of E-cadherin was assayed by Western blot;A correlation between E-cadherin mRNA levels and that of miR-148a in gastric cancer was investigated by Pearson correlation analyses;The relationship between aberrant methylation and E-cadherin expression was evaluated by treatment of gastric cancer cells with demethylation drug;The regulatory mechanism of miR-148a modulating E-cadherin was analyzed by transfection of gastric cancer cells with miR-148a mimics or DNA methyltransferase 1 (DNMT1) siRNA. Results:The mRNA and protein levels of E-cadherin were significantly downregulated in gastric cancer tissues compared with corresponding normal tissues,and were positively correlated with miR-148a expression. After treatment of MGC-803 and SGC-7901 cells with 5-aza-dC, a significant increase in E-cadherin mRNA and protein levels was observed. Moreover,overexpression of miR-148a,which had been verified to modulate DNMT1 expression, could reactivate the expression of E-cadherin. Knockdown of DNMT1 by DNMT1 siRNA could also increase the mRNA and protein levels of E-cadherin. Conclusion:The enforced expression of miR-148a may contribute to the reactivation of E-cadherin by suppression of DNMT1.

Abstract:Objective:To study early preventive effect of (-)-epigallocatechin-3-gallate (EGCG) on bone loss in ovariectomized (OVX) female rats. Method:Twelve-week-old female Sprague-Dawley rats,were divided into 3 groups:group A received intraperitoneal injection of 10 mg/(kg-d) EGCG 3 days after bilateral ovariectomy for 12 consecutive weeks;group B received bilateral ovariectomy alone;group C,received a sham operation. At the end of the experiment (24 weeks old),the tibias and the femurs were harvested for:① micro-CT scanning and measurement of bone mineral density (BMD) and bone morphological parameters:trabecular total bone volume (TV),bone volume,(BV),BV/TV,mean trabecular thickness (Tb.Th),mean trabecular number (Tb.N) and trabecular separation (Tb.Sp),cortical thickness,and cortical BV/TV;②The HE staining of bone tissues from female Sprague-Dawley (SD) rats. Results:According to the analysis of Micro-CT,the BMD and BV/TV of group A were significantly higher than the OVX group (P < 0.05). The trabecular separation (Tb.Sp) of group A was significantly lower than for group B (P < 0.05). There were no significant difference among 3 groups of the rest bone morphological parameters (P > 0.05). Bone histological studies indicated that trabecular bone was denser in group C,and the bone morphological status of group A was intermediate between groups A and C. Conclusion:EGCG had a positive effect on mitigating bone loss in OVX rats. Early stage supplementation of EGCG at a dose of 10 mg/(kg-d) after the onset of ovariectomy did not entirely eliminate bone loss.

Abstract:Objective:To investigate the effect of resveratrol on the proliferation and apoptosis of rat renal tubular epithelial cells (NRK-52E) and its possible mechanism. Methods: The NRK-52E cells were cultured in vitro. The NRK-52E cells were divided into four groups: normal glucose (5.6 mmol/L, the NG group), NG plus resveratrol (20 μmol/L, the NG+Res group), high glucose (30 mmol/L, the HG group),and HG plus resveratrol at 20 μmol/L (the HG+Res group). After pretreatment of resveratrol for 6 h and stimulation of HG for 24 h, the rate of cell proliferation was examined by the MTT method, the rate of cell apoptosis was determined by flow cytometry and the expression of GRP78 proteins in the NRK-52E cells was analyzed by Western blot. Results: Compared to the NG group, the rate of cell proliferation and apoptosis in the HG group was increased significantly. Simultaneous incubation with resveratrol inhibited NRK-52E cell proliferation and apoptosis (all P value < 0.01). As compared to the NG group, the expression of NOX4 and GRP78 protein in the HG group was significantly increased (all P value < 0.05). The expression of NOX4 and GRP78 protein in the HG+Res group was significantly decreased than that in the HG group (all P value < 0.05). Conclusion: This data demonstrate that resveratrol may exert antiproliferative and anti-apoptosis effects on rat renal tubular epithelial cells by inhibiting oxidative stress and endoplasmic reticulum stress.

Abstract:Objective:To study the regulatory effects and mechanisms of miR-143-5p on cadmium-induced apoptosis in LLC-PK1 cells. Methods:Microarray analysis was performed to detect dysregulated expression of miRNAs caused by cadmium. Reliability of microarray analysis was validated by quantative real-time PCR. Over-expression and low expression of miR-143-5p were simulated by its mimic and inhibitor transient transfection with Lipofectamine 2000,and the effect was verified by qRT-PCR. Hoechst 33258 staining and AnnexinV-FITC/PI method were used to detect apoptosis. Target gene of miR-143-5p was validated by bioinformatics analysis,real-time PCR and Western blot. Western blot was performed to analyze the regulatory effect of miR-143-5p on apoptosis-related pathway. Results:The expression of miR-143-5p was up-regulated by cadmium (P < 0.01). Compared with the negative control(miR-NC)group,the expression of miR-143-5p was significantly up-regulated after miR-143-5p mimic and down-regulated by inhibitor transfection (P < 0.01). Over-expression of miR-143-5p markedly increased LLC-PK1 cells apoptosis(P < 0.01). The mRNA and protein levels of AKT3 were both targeted and regulated by miR-143-5p. Over-expression of miR-143-5p reduced protein levels of p-Akt and p-Bad and increased expression of caspase-9 and caspase-3. Conclusion:MiR-143-5p may promote cadmium-induced apoptosis via targeting AKT3 and inhibiting Akt/Bad signal pathway in LLC-PK1 cells.

Abstract:Objective:To investigate the synoviolin expression in breast cancer tissues and its effects on the proliferation and migration of breast cancer cell line MCF-7. Methods:The expression of synoviolin in breast cancer tissues was detected by Western blotting and immunohistochemical staining. The proliferation and migration of MCF-7 cells were detected by CCK-8 kit assay, wound healing assay,and Transwell migration assay. The overexpression of synoviolin in MCF-7 cells was performed by adenoviral vector transfection. Results:The expression of synoviolin in breast cancer tissue was decreased than that of the corresponding peritumoral tissues;Overexpression of synoviolin inhibited the proliferation and migration as well as epithelial-mesenchymal transition (EMT) of the MCF-7 breast cancer cells. Conclusion:Down-regulation of synoviolin in patients with breast cancer may play a critical role in the proliferation and migration of breast cancer.

Abstract:Objective:To investigate the functions and mechanisms of EP3 receptor subtypes in promoting hepatoma Huh-7 cell growth and invasion. Methods:Human hepatocellular carcinoma Huh-7 cells were transiently transfected with EP3-4,EP3-5,EP3-6 and EP3-7 receptor. The expression of EP3 receptor in the transfected cells was detected by RT-PCR and Western blot. The cells were treated with various concentrations of selective EP3 agonist sulprostone. Cell viability was detected in the transfected cells by WST-8. Scratch test was performed to detect invasion ability in the transfected cells. Western blot was employed to detect the phosphorylation level of P-ERK. Results:The expression of EP3 receptor subtypes was markedly increased in the transfected cells by analysis of RT-PCR and Western blot. Under the treatment of 10 μmol/L sulprostone for 24 hours,the cell growth rate of Huh-7 cells transiently transfected with EP3-4,EP3-5 and EP3-7 receptor was increased to 126.7%,124.0%,and 123.8%,respectively. The growth of cell invasion rate separately came to 135.8%,123.5%,and 128.7%. The concentration of intracellular P-ERK in Huh-7 cells over-expressed of EP3-4,EP3-5 and EP3-7 receptor increased by 54.8%,33.4%,and 44.3% individually after 30 minutes exposure of 10 μmol/L sulprostone. However,over-expression of EP3-6 receptor cells had no significant difference in cell growth,cell invasion and the level of intracellular ERK phosphorylation. Conclusion:Our results suggest that EP3 receptor subtypes,including EP3-4,EP3-5 and EP3-7,mediate a great progress of the ability of proliferation and invasion in hepatocellular carcinoma Huh-7 cells,whereas EP3-6 receptor does not. Additionally,the function of EP3 receptor in accelerating cell growth and invasion of Huh-7 cells is in line with the activation of ERK.

Abstract:Objective:To investigate the effect of prostaglandin E2 (PGE2) on the invasion ability of ovarian cancer SKOV3 cell via upregulating the receptor of stromal cell-derived factor-1 (chemokine receptor 4,CXCR4). Methods:SKOV3 cells were treated with PGE2,EP1 receptor agonist,CXCR4 antagonist,EP1 receptor antagonist,protein kinase C (PKC) inhibitor and Ca2+ chelating agent. Real-time PCR,Western Blot and Transwell were performed to detect the levels of CXCR4 mRNA and CXCR4 protein as well as the invasion ability in SKOV3 cells. Results:The mRNA level of CXCR4 increased by 60.33% (P <0.01),while the level of CXCR4 protein increased by 122.88% (P < 0.01),and the invasion ability of SKOV3 cells increased by 112.24% (P < 0.01) after treated with 5 μmol/L PGE2. The invasion ability of SKOV3 cells decreased by 54.00% (P < 0.05) after treated with 10 μmol/L CXCR4 antagonist AMD3465 compared with the cells treated with PGE2. When treated with 5 μmol/L EP1 receptor agonist 17-PT-PGE2,the mRNA level of CXCR4 increased by 47.90 % (P <0.01),the level of CXCR4 protein increased 85.56% (P < 0.05),and the invasion ability of SKOV3 cells increased by 108.79% (P < 0.01). The protein level decreased by 54.86% (P < 0.05) and the invasion ability of SKOV3 cells decreased by 65.37% (P < 0.05) after treated with 10 μmol/L EP1 receptor antagonist sc-51322 compared with the cells treated with PGE2. When treated with 5 μmol/L PKC inhibitor BIS-1 and 10 μmol/L Ca2+ chelating agent BAPTA-AM,the protein levels of CXCR4 were decreased by 57.38%(P < 0.05) and 56.14% (P < 0.05) respectively,and the invasion ability of SKOV3 cells decreased by 52.63% (P < 0.05) and 55.26% (P < 0.05) respectively compared with the cells treated with 17-PT-PGE2. Conclusion:PGE2 might up-regulate the expression level of CXCR4 through EP1 receptor which could be partly related to the Gαq/Ca2+/PKC signaling pathway in ovarian cancer SKOV3 cells,and promote the tumor cells invasion.

Abstract:Objective:To determine the biological function of miR-122 in kidney tumors and the relationship between sprouty2 and miR-122. Methods:RT-qPCR was performed to measure the levels of miR-122 in 32 primary renal cell carcinoma (RCC) and their adjacent non-malignant tissue samples. The miR-122 inhibitor was successfully transfected into 786-0 and CAKI-1 cell lines. CCK-8 assays and cell cycle assays were performed to analyze the proliferation after transfection,respectively. The regulative function of miR-122 on sprouty2 was evaluated by Western blot assays. Dual luciferase reporting assays were performed to confirm whether sprouty2 is a direct target of miR-122. Results:In RCC specimens,miR-122 was significantly up-regulated (P < 0.05). The inhibition of miR-122 suppressed the proliferation (P < 0.05),induced cell cycle arrest (P < 0.05), and promoted the expression of sprouty2 protein levels (P < 0.05) in 786-0 and CAKI-1 cell lines. Sprouty2 was identified as a new target of miR-122 by dual luciferase,and fluorescence value was significantly increased (P < 0.05). Conclusion:miRNA promotes the proliferation of renal cancer cells and sprouty2 gene may be a target of miR-122. [Key words] miR-122;sprouty2;renal cell carcinoma;proliferation

Abstract:Objective:To construct the recombinant lentiviral vector containing LMP2A gene and measure the expression level of LMP2A in human carcinoma cell line of CNE1, as well as the effects of LMP2A on proliferation, migration and invasion on human carcinoma cell line of CNE1. Methods: The EB virus LMP2A fragment was amplified by PCR and subcloned into the lentiviral vetor plv by recombinant DNA technology. The resultant lentivirus was confirmed by PCR, restriction enzyme digestion and DNA sequencing. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pLMP2A and packaging plasmid pdelta-8.91 and pVSVG. CNE1 cells were infected by plv-LMP2A, and the expression of LMP2A in CNE1 cells was confirmed by RT-PCR, immunofluorescence and Western blot by screening of monoclonal. CCK8 assay, Transwell invasion assay and Wound-healing assay were performed to determine the effects of LMP2A expression on the proliferation, invasion and migration. Results: The recombinant lentiviral vector carried the LMP2A gene was successfully constructed. Results of RT-PCR, immunofluorescence and western blot indicated that CNE1 transgenetic cells could high express EBV-LMP2A. The LMP2A gene-transfected carcinoma cells grew vigorously and rapidly and up-regulated the ability of migration and invasion. Conclusion: The recombinant lentiviral vector pLMP2A was successfully constructed, and could be used to transfect human carcinoma cell line of CNE1. LMP2A could be highly expressed and can significantly promote CNE1 proliferation, migration and invasion, which laid a foundation of study on the biological function of Epstein-Barr virus and its potential role in gene therapy of tumors. [Key words] Epstein-Barr virus; latent membrane protein 2A; nasopharyngeal carcinoma; lentiviral vector

Abstract:Objective:CHOP is the standard regimen to treat diffuse large B cell lymphoma (DLBCL). With the anti-CD20 monoclonal antibody (rituximab,R) combined with CHOP regimen,the therapeutic effect of R-CHOP regimen is further improved. To investigate the clinical significances of treatment-related variables for the diffuse large B-cell lymphoma (DLBCL) patients who treated with front-line (R)-CHOP regimen. Methods:This retrospective study evaluated 52 conservative newly diagnosed patients who were treated with (R)-CHOP regimen. The prognostic roles of international prognostic index (IPI),average dose intensity per cycle (DIPC) and dose intensity per week (DIPW) of cyclophosphamide,doxorubicin,prednisone in (R)-CHOP regimen were evaluated. Results:The median follow-up period of this study was 34 months(from 6 to 95 months). Pearson Chi-Square analysis showed that complete response (CR) rates were significantly affected by average DIPW of prednisone <166.7 mg/m2,IPI scores and Ann Arbor stage (A or B group) (all P < 0.05). When the two factors were subsequently tested by multifactorial Logistic stepwise regression,average DIPW of prednisone <166.7 mg/m2 and IPI scores were again showed to be independent risk factors for CR rate. With regards to OS,it was showed to be significantly affected by IPI scores,average DIPW of cyclophosphamide <250 mg/m2 and doxorubicin <16.7 mg/m2 (all P < 0.05). In the subsequent Cox regression test,only IPI scores and average DIPW of doxorubicin <16.7 mg/m2 were showed to be independent prognostic factor for OS rate. Conclusion:In addition to IPI scores and Anna Arbor stage (A or B group),average DIPW of doxorubicin was also showed to be an important prognostic factor for the clinical outcome of newly diagnosed DLBCL patients who were first-line treated with (R)-CHOP regimen. It is strongly recommended to strictly adhere to planned (R)-CHOP regimen schedule in every possibility.

Abstract:Objective:To investigate changes of interlukin-1β (IL-1β) and prostaglandin E2 (PGE2) levels in gingival crevicular fluid (GCF) during distal movement of canines with Damon Q self-ligating brackets and traditional brackets,and to investigate the effect of Damon Q self-ligating brackets on the periodontium. Methods:Fifteen patients,aged 11-13 years and treated with maxillary first premolar extractions,participated in this study. The upper canines were bonded with self-ligating bracket and conventional MBT bracket,respectively. After leveling dentition, the upper canines were retracted with continuous forces of 100 g using nickel-titanium coil springs on 0.018 stainless steel archwire. GCF was collected from the distal site of each canine before activation and at 1 h,24 h,72 h,1 w,2 w,3 w and 4 w after initiation of the experiment. IL-1β and PGE2 concentrations were determined by enzyme-linked immunoadsordent assay (ELISA). Results:After 24 h of activation,the IL-1β in GCF of the self-ligating bracket group was significantly increased. At the self-ligating bracket group,the IL-1β and PGE2 levels in GCF were significantly higher than those of the traditional bracket group from the 72 h after the activation,and there were still significant differences at 4 w after activation. Conclusion:There were differences of IL-1β and PGE2 levels in GCF during distal movement of canines with Damon Q self-ligating brackets and traditional MBT brackets. Higher and more permanent biological effects were detected in the self-ligating group.

Abstract:Objective:To evaluate the effects produced by Headgear-Twin-block appliance for skeletal Class Ⅱ malocclusion. Method:One treatment group(Group R)consisted of 12 patients(8 males,4 females,mean age of 11 years 2 months)who treated with Headgear-Twin-block appliance.The other treatment group(Group T)also consisted of 12 patients(8 males,4 females,mean age of 11 years 6 months)who treated with Twin-block appliance without headgear. The control group(Group C)contained 12 patients(6 males,6 females,mean age of 10 years 8 months)giving up treatment after the examination. All the patients met the criteria as follows:(1)ANB≥4.5°;(2)Overjet≥6 mm and overbite≥3 mm;(3)Retrusive mandible. The patients of three groups were taken cephalometrics before and after the treatment or observation. Results:Compared -驻R with -驻T,SNA,U1-SN,Ls-E,Li-E,U1-Stms,U6-FHp decreased more and PP-SN,SN-MP,U6-CFH increased less,which had significantly difference (P < 0.05). Conclusion:Headgear-Twin-block appliance is effective to treat Class Ⅱ division 1 malocclusion by promoting the mandible and restraining the maxillary effectively to improve hard and soft tissue.

Abstract:Objective:To investigate the risk factors of acute intracerebral hemorrhage (ICH) early prognosis and provide a theoretical basis for clinical treatment. Methods:A total of 666 patients with ICH were collected and followed up the clinical outcomes for 21 days. Monovariant and multivariant logistic regression were performed to analyze the relationship between indicators,major treatment options of patients on admission and early prognosis. Results:Logistic regression analysis found that onset to admission time <6 hours (P = 0.003),suffering from high blood pressure on admission (P = 0.024),high blood glucose (P = 0.030),pulmonary infection (P = 0.035) and fever (P = 0.003),diagnostic score classification of stroke (P < 0.001) and bleeding sites (P = 0.032) were the risk factors in poor prognosis of ICH;the therapeutic plan of the combination of tradition Chinese and western medicine was the protective factor for ICH. Conclusion:The predictor of early prognosis by integrating multiple risk factors associated with acute ICH was beneficial for the implementation of individual clinical decision to reduce patients＇ mortality and raise their quality of life.

Abstract:Objective:To screen the peptides biomarker from sera of the newly-developed advanced schistosomiasis (NDAS) patients and to lay the foundation for schistosomiasis diagnosis and pathogenesis. Methods:Peptides were collected from the sera of NDAS patients and health control by MB-WCX kit (Bruker Daltonics GmBH),followed by analysis with Flex-Analysis,ClinProTool algorithm and MASCOT using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Results:In the mass range from 0.8 to 13 ku,7 peaks with a clear difference in amplitude between the NDAS patients and control groups were detected,5 peaks with mass charge ratio (m/z) of 2 661,2 991,3 241,5 337 and 5 906 were down-regulated and 2 peaks with a m/z of 2 082,4 282 were up-regulated in NDAS patients. Furthermore,m/z of 2 662,4 209,5 337,5 906 were down-regulated and 2 082 was up-regulated in advanced schistosomiasis patients. Conclusion:m/z 3 241,4 282 could give a potential application value for NDAS patients diagnosis and lay the foundation for schistosomiasis pathogenesis.

Abstract:Objective:To investigate the changing of regional homogeneity (ReHo) and its relationship with the severity of the symptom in untreated obsessive-compulsive disorder (OCD) patients using resting-state functional magnetic resonance imaging (RS-fMRI) technology. Methods: A total of 46 obsessive-compulsive disorder patients and 31 age-sex-education matched healthy adults were scanned with RS-fMRI for ReHo analysis using SPM8, DPARSF and REST toolbox. The Yale-Brown Obsessive Compulsive Scale (Y-BOCS), 17-Item Hamilton Rating Scale for Depression (HAMD-17) and Hamilton Anxiety Rating Scale (HAMA) were performed to assess the clinical symptoms. And then, the relationship between the changing of ReHo and severity of clinical symptoms was determined. Results: Compared with the controls, the patients showed higher ReHo in the left prefrontal lobe, and lower in the left occipitotemporal regions and bilateral fusiform gyrus. However, no significant correlation was found between the different areas and the clinical symptom scores. Conclusion: Resting-state fMRI witnessed abnormal brain areas activity in untreated obsessive-compulsive disorder patients. The functional changes suggest that the abnormal synchrony of brain regional shortage functional connectivity probably plays an important role pathophysiologically.

Abstract:Objective:This paper proposed an improved K-means clustering algorithm based on kernel density estimation which is used for the automatic segmentation of whole-body bone scan image. Methods:First,we sharpened and smoothed the 2D SPECT whole-body scan image for preprocessing. Second,we used Gaussian kernel density estimation curve to obtain optimal clustering centers as the initial value of K-means clustering algorithm. And then,we segmented the image using the K-means clustering algorithm. Finally,the template match method was performed to delete wrong recognized areas. Results:Image preprocessing provided clearer and more detailed activity structures and improve image quality. The improved K-means clustering algorithm generated a higher degree of Tanimoto similarity than traditional K-means method,and it had a less running time than others. Conclusion:Kernel density estimation can effectively avoid the blindness of the initial clustering center selection in the K-means method,and make the clustering results more rapid,accurate and stable. The proposed algorithm is suitable for whole-body bone scan image segmentation which has important significance to the analysis of region of interest.

Abstract:Objective:To study the alterations of energy metabolism and oxidative stress in mouse liver after infection of recombinant adenovirus. Methods:Seven-week-old male Balb/c mice were randomly assigned to control and adenovirus infection groups. Recombinant adenovirus encoding green fluorescent protein (GFP) or phosphate-buffered saline (PBS) was delivered by tail vein injection. Tissue distribution of GFP was determined by fluorescence microscopy. The levels of alanine transaminase (ALT),aspartate amino transferase (AST),glucose,cholesterol,triglyceride,β-hydroxybutyrate (β-HB),fibroblast growth factor-21 (FGF-21) and malondialdehyde (MDA) in plasma as well as hepatic glucogen contents were detected 5 days after injection. Hepatic expressions of genes involved in gluconeogenesis,lipogenesis and fatty acid oxidation were examined by quantitative real-time PCR. To determine the oxidative stress levels in mice,MDA in blood and hepatic mRNA expression levels of reactive oxygen species (ROS)-generating or antioxidant genes were also examined. Results:The expression of GFP delivered by tail vein injection was mainly in the liver of mice,with the highest levels at 5 days after injection. Compared with the control mice,adenovirus infection led to an obvious increase of ALT and AST in plasma of mice in the adenovirus infection group. The levels of β-HB and FGF-21 were also elevated. Analysis of the liver transcripts indicated that genes implicated in fatty acid oxidation were increased while those involved in lipogenesis and gluconeogenesis pathways were suppressed by adenovirus infection. Besides,hepatic levels of ROS were also increased. Conclusion:Our findings indicate that adenovirus infection improved fatty acid oxidation and inhibited gluconeogenesis and lipogenesis. The alterations of energy metabolism may be mediated by the increased oxidative stress in liver after adenovirus infection. [Key words] recombinant adenovirus;oxidative stress;gluconeogenesis;fatty acid oxidation

Abstract:Objective:To investigate the effect of interleukin-17 (IL-17) on rat renal fibroblast NRK-49F transforming into myofibroblast and expressing extracellular matrix (ECM) as well as the signal mechanism involved. Methods: Western blot detected the effect of IL-17 on NRK-49F cells which was induced by transforming growth factor-β (TGF-β) transforming into myofibroblasts and expressing alpha-smooth muscle actin (α-SMA) and I-collagen and fibronectin. The mRNA expression levels of ECM I-collagen and fibronectin as well as regulatory effect of IL-17 in NRK-49F cells were detected by real-time PCR after 24, 48 and 72 h, respectively. Western blot was performed to detect classical Smad-dependent and Smad-independent signaling of TGF-β and its phosphorylation level, in order to explain molecular mechanism of IL-17 on regulating NRK-49F. Results: IL-17 inhibited NRK-49F cell line transforming into myfibroblast and expressing α-SMA, I-collagen and fibronectin. IL-17 had no effects on the expression of classic Smad signaling molecule, however, it inhibited the activation of Akt-TSC2-p70S6K through Smad-independent signaling pathway. Conclusion: IL-17 inhibits NRK-49F cell line transforming into myfibroblast and expressing α-SMA, I-collagen,and fibronectin by blocking the activation of Akt-TSC2-p70S6K through Smad-independent signaling pathway.

Abstract:Objective:To investigate the effects of silent information regulator 1 (Sirt1) over-expression on high glucose-induced proliferation and transforming growth factor-β1 (TGF-β1) expression in human glomerular mesangial cells (HMCs). Methods: HMCs were cultured in vitro. The HMCs were transfected with either lentiviral vectors containing the Sirt1 cDNA (LV-Sirt1) or empty vectors (LV-CTL). The expression of Sirt1 protein was detected by Western blot. Those HMCs transfected with lentiviral vectors were divided into 4 groups including normal glucose (5.5 mmol/L) plus LV-CTL (NG+LV-CTL), high glucose (30 mmol/L) plus LV-CTL (HG+ LV-CTL), normal glucose plus LV- Sirt1 (NG+LV-Sirt1), and high glucose plus LV-Sirt1 (HG+LV-Sirt1) groups. Those untransfected HMCs were cultured at normal glucose (NG), normal glucose plus mannitol (5.5 mmol/L glucose + 24.5 mmol/L mannitol ) (NM) or high glucose (30mmol/L) (HG) as control. After treatment for different periods, the cell proliferation of each group was examined using CCK-8 detection kit, and the levels of mRNA and protein of TGF-β1 from each group were measured through real-time polymerase chain reaction (PCR) and Western blot, respectively. Results: The HMCs, which was stably over-expressed Sirt1 protein, was obtained after recombinant lentivirus infection and puromycin selection. Compared with the NG group, the proliferation levels of HMCs from the HG, HG+LV-CTL and HG+LV-Sirt1 groups on 12, 24 and 48 hours were increased significantly (each P < 0.01). However, the proliferation level of the HG+LV-Sirt1 group is significantly lower than that in the HG and HG+ LV-CTL groups. As compared to counterparts of the HG and HG+ LV-CTL groups, the levels of mRNA and protein of TGF-β1 from the HG+LV Sirt1 group were decreased significantly (each P < 0.01). The levels of mRNA and protein of TGF-β1 from the HG treated groups were higher than those of the NG group (each P < 0.01). In addition, there were no difference on the levels of mRNA and protein of TGF-β1 among NG, NM, NG+LV-CTL and NG+LV Sirt1 groups (each P > 0.05). Conclusion: Sirt1 over-expression can inhibit cell proliferation and TGF-β1 expression of HMCs induced by high glucose treatment. Sirt1 gene may become a new therapeutic target of diabetic nephropathy.

Abstract:Objective:To study the short-term clinical efficacy and safety in patients undergoing percutaneous coronary rotational atherectomy or percutaneous coronary intervention for severe coronary calcification. Methods: A total of 112 patients with coronary artery intervention treatment that confirmed as severe calcification by coronary angiography were divided into the PTCRA group and the non-PTCRA group. The perioperative complications,mortality and occurrence rates of the major adverse cardiovascular events (MACE) were collected postoperative 9-12 months. Results: There was no statistical difference (P > 0.05) between the two groups on age,sex,hypertension,smoking,diabetes,cholesterol,high-density lipoprotein cholesterol (HDL-c),low density lipoprotein cholesterol (LDL-c),triglycerides,eGFR,heart ejection fraction (EF),and other factors. The mortality and occurrence rates of MACE between the two groups had no statistical difference (P > 0.05):and the PTCRA group had less complications than the non-PTCRA group (P < 0.05). Conclusion: Percutaneous coronary rotational atherectomy with percutaneous coronary intervention can effectively reduce perioperative complications. The treatment for complexity,calcified lesions is safe and effective,and has a good short-term clinical efficacy.