Desmoplastic small round cell tumors (DSRCTs) are a rare malignancy found in male adolescents that initially occur mostly in the abdominal cavity. Diagnosis is based on the histologic analysis of biopsies, which typically show small round blue cells in nests separated by abundant desmoplastic stroma. DSRCTs are associated with a unique chromosomal translocation t (11:22) (p 13;q 12) that involves the Ewing's sarcoma (EWS) gene and the Wilms' tumor (WT1) gene. Reverse transcriptase-polymerase chain reaction can be used to detect the fusion gene in fresh or paraffin-embedded tissues, which confirms the diagnosis. The prognosis is particularly poor. The median survival ranges from 17 to 25 months. Management of DSRCT remains challenging despite the use of aggressive ther-apies such as polychemotherapy, debulking surgery, and whole abdominal radiation. Several methods for improving patient survival are being evaluated, such as the addition of chemotherapy and targeted therapies to normal neoadjuvant protocols, complete surgical resec-tion with hyperthermic intraperitoneal chemotherapy, postoperative intensity-modulated radiation therapy, and yttrium-90 microsphere liver embolization for treating hepatic metastases.

Objective Myocardial infarction and subsequent heart failure remain the most dominant health challenges worldwide.Therapeutic angiogenesis has emerged as a potential novel treatment for severe ischemic heart disease and there is increasing evidence that cell transplantation may improve the perfusion and contractility of myocardium in animal models.This study was designed to examine the endothelial growth potential and whether transplantation of human umbilical cord derived mesenchymal stem cells can improve local blood flow in a mouse ischemic hindlimb model.Methods The mesenchymal stem cells derived from human umbilical cord of passage 5 were differentiated in an endothelial differentiation medium containing vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in vitro.Samples were observed for 2 weeks.The human umbilical cord derived mesenchymal stem cells were transplanted into a hindlimb ischemia mouse model in vivo.Four weeks later,immunofluence was used to identify the migration and differentiation of the transplanted cells towards endothelial linage.Laser Doppler perfusion image was used to evaluate the local blood flow of the hindlimb.Results Results After incubation with VEGF and bFGF,the human umbilical cord derived mesenchymal stem cells started to form interconnected clusters and a network was formed.Four weeks after transplantation,the transplanted cells were sprouting f0rom the local injection and differentiated into endothelial cells,contributed to the recovery of local blood flow obviously as compared with control group.Conclusion Human umbilical cord derived mesenchymal stem cells have the ability to differentiate into endothelial cells,contribute to the local angiogenesis in a hindlimb ischemia mouse model and represent a new source for therapeutic angiogenesis for clinical applications.

BACKGROUND: Mesenchymal stem cells (MSCs) are an important component of the in vivo microenvironment and act on multiple biological behaviors of tumor cells. The potential clinical value of MSCs has become an issue of concern in recent years.OBJECTIVE: To investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with umbilical cord-derived MSCs (UC-MSCs) using cDNA microarray.METHODS: In vitro co-culture system was constructed, and then cellular proliferation, apoptosis and differentiation status of NB4 cells treated with UC-MSCs were evaluated. Two cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without UC-MSCs. The probes were labeled with fluorescence dyes individually, hybridized with cDNA microarray, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in two gene expression profiles.RESULTS AND CONCLUSION: UC-MSCs promoted the proliferation and differentiation, while reduced the apoptosis of NB4 cells. The analysis of gene expression profiles indicated that after co-culture with UC-MSCs, 530 genes were up-regulated and 53 genes were down-regulated. Accordingly, specific gene function and pathway signaling related were also regulated to some extent. Overall, UC-MSCs influence can major biological behaviors of NB4 cells by changing expression of a large amount of genes, gene-related function and multiple intracellular signaling pathways.

Objective To explore the relationship between the Interleukin 6 , matrix metalloproteinases 2 and early embryo arrest. Methods Real time-PCR was used to measure the mRNA expression of IL-6 and MMP-2 and immunohistochemistry (IHC, SP method)was used to measure the location and expression of the two different kinds of protein in villus. ELISA was used to measure the level of IL-6 in serum. Results Real-time PCR and IHC showed that the expression levels of IL-6 was significantly lower in experimental group than in control group, and MMP-2 was significantly higher than the control group (P < 0.05). Differenc of IL-6 level in serum between the two groups was not statistically significant (P > 0.05). Conclusion Proper expressions of IL-6 and MMP-2 in the villus tissue play a key role in the maintenance of early pregnancy.

ObjectiveToll-like receptors (TLRs) play important role in the progression and tumor immunity of some types of cancer,some research have demonstrated that agonist of TLR3 can trigger apoptosis of cancers.This study was proposed to investigate if Poly(I:C),the specific agonist of TLR3,could impact proliferation or apoptosis of progressive breast cancer cells MDA-MB-231,and to investigate the primary mechanism of the function.MethodsExpression of TLR1-10 mRNA was detected by quantitative real-time reverse transcription-polymerase chain reaction.Cell Counting Kit-8 was used to determine the inhibitory effect of Poly(I:C) on proliferation of MDA-MB-231 cells.Cell apoptosis was assayed by flow cytometry with V-FITC/PI staining.Results First,the toll-like receptors 1-10 were all expressed on MDA-MB-231 cells,while the expression level of TLR8 was lower than that of others.Second,according to the CCK-8,the proliferation of MDA-MB-231 cells was inhibited,but the apoptosis was not affected on the basis of Apoptosis Kit.At last,the mRNA expression of TNF-α、IFN-β and IFN-γ were elevated approximately 20 times after Poly(I:C) stimulation for 6 hours.ConclusionMDA-MB-231 cells express all toll-like receptors on mRNA level,and TLR8 was expressed lower than others.The stimulation of TLR3 with Poly(I:C) can inhibit the proliferation of MDA-MB-231,but had no effect on apoptosis.TNF-α、IFN-β and IFN-γ maybe participate in this process.

Objective:To investigate the expression levels and clinical significance of glioma-associated oncogene homolog 1 (GLI1) and sonic hedgehog signaling molecule (Shh) in the malignant transformation of ovarian endometriosis (EM).Methods:The expressions of GLI1 and Shh were detected by real-time reverse transcription (RT)-polymerase chain reaction (PCR) and EnVision method in 50 cases of ovarian EM tissues, 35 cases of atypical endometriosis (aEM) and 50 cases of endometriosis-associated ovarian cancer (EAOC). The expression differences of two molecular markers in the malignant transformation of ovarian EM were compared, and the relationships between two molecular markers and the clinicopathological features and prognosis of EAOC were analyzed.Results:(1) RT-PCR showed that the expression levels of GLI1 mRNA in EM, aEM and EAOC group were 1.77±0.40, 3.54±0.44, and 7.80±0.24, respectively. The expression levels of Shh mRNA were 0.95±0.21, 3.14±0.35, and 5.41±0.31, respectively. GLI1 and Shh mRNA in EAOC group were significantly higher than those in EM and aEM group (all P<0.01), and there were statistically significant differences between EM and aEM group (all P<0.01). The percentages of GLI1 in ovarian EM, aEM and EAOC were 32% (16/50), 57% (20/35), and 66% (33/50), respectively, meanwhile, the positive expression rates of Shh were 20% (10/50), 49% (17/35), and 54% (27/50), respectively (all P<0.01). GLI1 mRNA expression was positively correlated with Shh mRNA expression in EAOC tissues ( r=0.721, P<0.01). The expressions of GLI1 protein were proportionated to Shh protein in EAOC tissues ( r=0.608, P=0.001). (2) The expression of GLI1 was significantly related to the International Federation of Gynecology and Obstetrics (FIGO) stage, cancer antigen 125 (CA 125) levels, lymph node metastasis, and Platinum resistance in EAOC patients (all P<0.05). The expression of Shh were related to FIGO stage and lymph node metastasis in EAOC patients (all P<0.05). Logistic regression analysis showed that GLI1 expression was an independent risk factor for poor prognosis in EAOC patients ( P<0.05). Kaplan-meier survival analysis showed that the overall survival rate of EAOC patients with high GLI1 expression and low GLI1 expression was 12.1% and 35.3%, respectively, with statistical significance ( χ2=10.73, P<0.01). The overall survival rate of EAOC patients with high and low expression of Shh protein was 11.1% and 30.4%, in which there was statistically significant difference ( χ2=3.96, P=0.047). Conclusion:GLI1 and Shh are highly associated with the malignant transformation of ovarian EM, which may play a role in promoting malignant degeneration of ovarian EM, and the high expression of GLI1 and Shh indicates a poor prognosis in EAOC patients.

Objective:To establish an animal model of chronic systemic inflammation with long-term high expression of circulating IL-6 by introducing exogenous IL-6 gene transfer vector.Methods:Recombinant murine IL-6-encoding adeno-associated virus (AAV-IL-6) was constructed. Twenty-one 24-week-old male C57BL/6J mice were randomly divided into three groups with seven in each group: AAV-IL-6 group, vector control (AAV-ctrl) group and blank control group. At 0, 8 and 16 weeks of intervention, the mice in the three groups were injected with AAV-IL-6 (100 μl 0.5×10 10 vp/ml), unloaded AAV (100 μl 0.5×10 10 vp/ml) and the same volume of saline in the tail vein, respectively. IL-6 levels in mouse serum were measured by ELISA. The general condition of mice was observed and blood routine tests were performed. Changes in blood biochemical parameters and C-reactive protein (CRP) levels were detected. At the end of 24-week intervention, the mice were sacrificed and the myocardium, liver, spleen, quadriceps femoris, knee joint and middle femur were taken for HE staining. Results:At 4, 8, 16 and 24 weeks after intervention, serum IL-6 levels were (75.41-169.28) pg/ml in the AAV-IL-6 group, while in the two control groups, the levels were below the lower limit of detection (7.8 pg/ml). At 24 weeks after intervention, the body weight of mice in the AAV-IL-6 group was significantly lower than that of mice in the two control groups; the neutrophil counts and CRP level in the AAV-IL-6 group were higher than those in the two control groups, while the levels of albumin, creatinine, triglyceride and cholesterol were lower than those in the two control groups. There were no differences in the aforementioned parameters between the two control groups. Compared with the blank control group, both AAV-IL-6 and AAV-ctrl groups showed increased lymphocyte counts. All mice had normal liver and kidney functions at the end of intervention. Histopathological findings indicated that the mice in the AAV-IL-6 group had focal infiltration of lymphocytes in the central venous area of the liver and around the myocardial and the skeletal muscle fibers, diffuse infiltration of multinucleated giant cells in the spleen, atrophic skeletal muscle, disorganized growth plate, reduced chondrocyte hypertrophic zone, thinner bone cortex and trabecular, and reduced osteoid. There were no histopathological changes in mice of the two control groups.Conclusions:Repeated tail vein injection of AAV-IL-6 could achieve long-term high expression of circulating IL-6 in mice, which manifested the phenotype of chronic systemic inflammation in preliminary detection and provided a safe, effective and simply accessible animal model for related studies.

Objective To study the effects of heat shock treatment of rat bone marrow mesenehymal stem cells(MSCs),the apoptosis ratio of treated-cells under low serum condition and the treated-cells transplantation on left ventricular function in rats with myocardiaIinfarction.Methods MSC8 were heat-treated under 42℃for 30 min,then the heat shock protein-70(HSP-70)was detected bv Western blot.The apoptosis ratio of heat-treated MSCs under low serum condition was tested by Annexin kit.The treated-MSCs labeled with Dil were transplanted into infarcted myocardium and 8 weeks later,the cardiac function of rats in each group was evaluated by echocardiography and cardiac catheterization.Results The immunophenotype of heat-treated MSCs did not vary,Western blot confirmed a higher level expression of HSP-70 in the treated-MSCs group as compared with that in the control group.The early apoptosis ratio was lower in treated-MSCs measured by flow cytometry with annexin staining than that of MSCs when cultured with low serum medium.After 8 weeks,LVEF,LVSP,+dp/dtmax,and-dp/dtmax were significantly higher,and the LVEDP was significantly lowar in heat-treated MSCs transplantation group than that in the control group.Conclusions Heat shock pretreatment of MSCs enhances the tolerance of MSCs to low serum medium,whereas does not lcad to the change of the cell immunophenotype.Transplantation of heattreated MSCs might improve the cardiac function in a rat myocardialinfarction model.

Objective To investigate the impact of high mobility group box1 and GRACE score on the clinical prognosis of patients with acute coronary syndrome undergoing selective percutaneous coronary intervention. Methods A total of 380 consecutive patients initially diagnosed with acute coronary syndrome undergoing selec-tive PCI between January 2014 and March 2015 were included,with 200 of them assigned into low high mobility group box1(HMGB1<445 ng/mL)and the other 180 patients into high mobility group box1(HMGB1≥445 ng/mL).The baseline characteristics and laboratory indexes were collected on admission,GRACE score were calculat-ed at admission.The difference between the high and low high mobility group box1 were analzyed and the influenc-ing factors of patients with acute coronary syndrome undergoing selective percutaneous coronary intervention were studied. The mean follow-up period was 24 months,and the clinical end points were deaths from various causes and readmission for coronary heart disease. Results There were significantly differences statistically between the groups of high and low high mobility group box1 in clinical diagnosis. lipoprotein associated phospholipase A2, GRACE score,mean platelet volume,red cell distribution width,age,and left ventricular ejection fraction(P <0.05). The correlation analysis showed that HMGB1 was significantly related to lipoprotein associated phospholi-pase A2 and GRACE score,with the correlation coefficents of 0.575,0.836,respectively(P<0.05).COX analy-sis showed that HMGB1,lipoprotein associated phospholipase A2,GRACE score had statistical significance for survival outcomes(P<0.05),and the area under the ROC curve drawn by combining the three was 0.851(95% CI 0.811 ~ 0.891,P < 0.05). Conclusion There was a good correlation between HMGB1 and GRACE score. HMGB1 is a good predictor of clinical outcomes in the patients with acute coronary syndromes undergoing elective PCI treatment.

Objective:To evaluate the clinical value of 5.0 T ultra-high filed MRI system in assessing intracranial arteries segments and vessel branchers.Methods:This study was a prospective study. Totally 40 consecutive healthy volunteers were recruited from Zhongshan Hospital, Fudan University from September 1, 2021 to November 30, and all participants who underwent either 3.0 T or 5.0 T time-of-flight MR angiography (TOF-MRA) in random order were divided into 3.0 T MR group and 5.0 T MR group with 20 volunteers for each group. Image quality was assessed by Likert 5 scoring systems and signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR),and score in visualization of intracranial arteries [middle cerebral artery (MCA) and its segments, anterior cerebral artery (ACA) and its segments, posterior cerebral artery (PCA) and its segments, lenticulostriate arteries (LA) and pontine artery (PA)] were assessed from 0 to 3 (≥2: good depiction of vessel segment). Quantitative indicators were compared between 2 groups using independent t test or Mann-Whitney U test. Results:Among the 40 subjects, there were 29 males and 11 females, aged 20-69 (50±12) years. SNR and CNR were both significantly higher in 5.0 T MR group than those in 3.0 T MR group (SNR: 187±9 vs 91±4, t=31.59, P<0.001; CNR: 156±7 vs 70±4, t=31.45, P<0.001), but there was no significant difference in subjective scores of image quality between the 5.0 T MR and 3.0 T MR groups [5.0 (4.0, 5.0), 5.0 (5.0, 5.0) points, respectively, Z=-1.23, P=0.218]. In the evaluation of cerebral arteries, the visualizations of the proximal and middle segments of MCA, ACA and PCA was better than those in the 3.0 T MR group, and there was no significant difference in the scores ( P>0.05), while the visualizations of proximal arteries in the 5.0 T MR group were significantly better than those in the 3.0 T MR group ( P<0.05). Furthermore, small vessel branches such as LA and PA in 5.0 T MR group were visualized better than those in 3.0 T MR group ( P<0.001). Conclusion:TOF-MRA by ultra-high filed 5.0-T provides an optimal choice in visualization of distal large arteries and small vessel branches, which could be useful for the diagnosis on cerebral vascular disease.