[Objective]To explore effect of polypeptides from chlamys farreri(PCF) on human umbilical cord blood cells(HUCBCs) differentiation in vitro,the effects of the extract on protein tyrosine kinase(PTKs) was studied in vitro.[Method]Getting human umbilical cord blood cells and cultured with the PCF at dilution concentrations of 1:1600,1:3200,1:6400,1:12800 nerve growth factor(NGF) was used as positive control and DMEM as negative one.After 48,72,96 h,the HUCBCs was cultured and the activity of protein tyrosine kinase(PTK) was examined by ATP in corporation test using PtK assay system.[Result]HUCBCs in the negative control group had no PTK activity,but that of positive control group was detected significantly.The PTK activity in extract of polypeptides from chlamys farreri groups increased at 48h(P

Objective To observe the therapeutic efficacy and safety of the glucagon-like peptide-1 ( GLP-1 )analogue combined with isulin for treating obese or overweight type 2 diabetes.Methods 40 cases with obese or overweight type 2 diabetic patients( body mass index(BMI) ≥24) who have previously used human insulin,and experienced poor glycemic control( ≥7.5％ ) were selected.They were randomly divided into two groups.A group treated by GLP-1 analogue liraglutide plus insulin,and B group by continuous adjustment of insulin.12 weeks later,fasting blood glucose(FBG),2-hour plasma glucose (2hPG),hemoglobin A 1 C (HbA1c),body weight were observed and the incidence of hypoglycemia,insulin dosage were recorded.Results Weight and insulin dosage of group A was significantly lower than those of group B after treatment 12 week( t =2.738,2.865,all P ＜ 0.01 ).Numbers of hypoglycemia events were 6 in group A(30％ ),and 9 in group B(45％ ),but there was no statistical significance between the two groups ( P ＞ 0.05).Conclusion The addition of liraglutide to insulin in patients with obese or overweight type 2 diabetes is associated with reductions in weight and insulin dosage,without increase risk of hypoglycemia.This treatment proved effective and safe.

Objective:To explore the relationship of lymphocyte function-associated antigen 3 (LFA-3, CD58) expression in glioma with clinical features and its role in clinical prognosis. Methods:Clinical data and mRNA microarray data of 514 patients with glioma were obtained from The Cancer Genome Atlas and were used to study the expression of LFA-3 (CD58). Cox regression was used to analyze the relationship between CD58 expression and survival of patients with glioma. Multivariate analysis of variance was used to further analyze the relationship of CD58 with age, sex, and pathological grade of glioma. Results:The results of the stratified χ2 test of CD58 expression and tumor grading were shown, considering tumor type, gender and age of diagnosis (all P<0.05). CD58 expression was sig-nificantly correlated with overall survival (OS) and disease-free survival (DFS) of patients with glioma. Patients with high CD58 expres-sion presented short OS and DFS (P<0.0001). Conclusion:CD58 possibly promotes tumorigenesis in gliomas and thus can serve as a potential tumor diagnostic marker and individual therapeutic target.

BACKGROUND:Previous studies have indicated that ulinastatin can inhibit RANKL-induced osteoclastogenesis on RAW264.7 cells and also lower matrix metal oproteinase-9 expression and activity. However, it remains be unclear whether ulinastatin has the intervention effect on polymethyl methacrylate (PMMA)-induced MC3T3-E1 mouse preosteoblast apoptosis or not. ＜br> OBJECTIVE:To explore the intervention role of ulinastatin on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis and its effects on type I col agen, osteocalcin, matrix metal oproteinase-2 mRNA expression. ＜br> METHODS:MC3T3-E1 mouse preosteoblasts at passages 6 and 7 were divided into four groups:blank group (only cultured MC3T3-E1 mouse preosteoblast), PMMA-induced group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension), low dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+500 U/mL ulinastatin) and high dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+5 000 U/mL ulinastatin). MTT method was adopted to detect the proliferation activity of proliferative activity of MC3T3-E1 mouse preosteoblast;alizarin red staining method was used to observe mineralization nodules of MC3T3-E1 mouse preosteoblast among different groups;the change of apoptosis rate for MC3T3-E1 cells was detected by flow cytometry analysis;semi-quantitative RT-PCR was taken to analyze type I col agen, osteocalcin, matrix metal oproteinase-2 mRNA expression level in MC3T3-E1 mouse preosteoblasts among different groups. ＜br> RESULTS AND CONCLUSION:Compared with the blank group, PMMA significantly inhibited the proliferation activity of MC3T3-E1 mouse preosteoblast (P＜0.05), and however significantly promoted cells apoptosis (P＜0.05). After addition of different concentrations of ulinastatin (500, 5 000 U/mL), the proliferation activity of MC3T3-E1 mouse preosteoblasts significantly raised (P＜0.05), and cells apoptosis rate significantly decreased (P＜0.05), showing the dose and time-dependent relation. Type I col agen and osteocalcin mRNA expression levels both significantly decreased after co-culture in PMMA group compared with the blank group (P＜0.05), matrix metal oproteinase-2 mRNA expression level, however, significantly increased (P＜0.05). After intervention with 5000 U/mL ulinastatin, type I col agen and osteocalcin mRNA expression levels both significantly increased, while matrix metal oproteinase-2 mRNA expression level significantly decreased (P＜0.05). PMMA group showed no obvious mineralization nodules. Yet, mineralization nodules were formed in the blank group, high and low dose ulinastatin groups. These results indicate that ulinastatin could have the inhibitory effect on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis, and it could promote type I col agen and osteocalcin mRNA expression and yet suppress matrix metal oproteinase-2 mRNA expression.

BACKGROUND:It is presumed that urinary trypsin inhibitor could have protective effects on local and systemic tissues and could inhibit osteoclast proliferation and activation under long-term chronic inflammation conditions and in ischemic and anoxic environment which was induced by prosthetic wear. OBJECTIVE:To investigate the inhibitory effect of ulinastatin on receptor activator for nuclear factor-κb ligand-induced differentiation, proliferation and osteoclastogenesis of RAW264.7 cells and its effects on matrix metal oproteinase-2, matrix metal oproteinase-9 expression level and activity. METHODS:Mouse monocyte/macrophage cellline RAW264.7 was treated with different concentrations of urinary trypsin inhibitor (0, 500, 5 000 U/mL) for 24, 48 and 72 hours. Experiments were divided into four groups:the blank group (RAW264.7 cells), receptor activator for nuclear factor-κb ligand-induced group (0 U/mL ulinastatin), 500 U/mL ulinastatin group and 5 000 U/mL ulinastatin group. RESULTS AND CONCLUSION:(1) MTT results indicated that there was no significant difference on the proliferation of RAW264.7 cells treated with urinary trypsin inhibitor at 0-5 000 U/mL (P>0.05) (2) Tartrate-resistant acid phosphatase staining results revealed that compared with receptor activator for nuclear factor-κb ligand-induced group, the number of tartrate-resistant acid phosphatase-positive cells was significantly less in the ulinastatin group (P<0.05), showing a time-dose dependent manner. (3) Immunohistochemisical results found that compared with receptor activator for nuclear factor-κb ligand-induced group, the percentage of matrix metal oproteinase-9-positive cells was apparently lower in the ulinastatin group. (4) Western blot assay results demonstrated that matrix metal oproteinase-9 expression was low in the RAW264.7 cells alone. At 48 hours after addition of receptor activator for nuclear factor-κb ligand, matrix metal oproteinase-9 protein expression was large. At 72 hours after culture in the 5 000 U/mL ulinastatin group, matrix metal oproteinase-9 protein expression was evidently reduced. (5) Gelatin zymography results showed that compared with the receptor activator for nuclear factor-κb ligand-induced group, matrix metal oproteinase-9 expression was significantly lower in the 5 000 U/mL ulinastatin group (P<0.05). Results suggested that urinary trypsin inhibitor inhibited receptor activator for nuclear factor-κb ligand-induced osteoclastogenesis and diminished matrix metal oproteinase-9 expression and activity.

Objective To observe the clinical efficacy and safety of second transurethral resection combined with instillation therapy and transfusion therapy of dendritic cells pulsed with tumor cells on non muscle-invasive bladder cancer.MethodsEighty patients with stage T1 non muscle-invasive bladder cancer were included in this protocol in which all patients prospectively received second transurethral resection within 4 to 6 weeks following initial resection.All 80 cases were divided into a DC group and a control group.In the DC group,dendritic cells pulsed with tumor cells were transfused between 6 -8 weeks.Bladder instillation therapy and follow-up was applied on the control group.The recurrence rate,the clinical efficacy and adverse reactions were observed and compared between the two groups.ResultsIn the initial resection,21.3％,67.5％ and 11.2％ had G1,G2 and G3 transitional cell carcinoma,respectively.Twenty-seven (33.7％) had residual tumors at the second TUR,8 patients had Ta(29.6％ ) and 19 had T1 (70.4％).After the initial TUR-Bt,residual tumors were detected in 11.1％,70.4％ and 18.5％ in G1,G2 and G3,respectively.In the 8 Ta cases,2 cases moved to a higher grade,while the grade was unchanged in 6 cases.In the 19 cases with stage T1,12 had a higher grade,5 had a lower grade and 2 remained the same.In the DC group,5 cases suffered chills and fever when dendritic cells were transfused.The fever was releaved when dexamethasone was administered.The white blood cells count,creatinine and alanine aminotransferase had no statistically significance change at pre-therapy,one year after therapy and two years after therapy (P ＞0.05).The index of CD4 、CD8 、CD4/CD8 had statistically significance change at pre-therapy,one year after therapy and two years after therapy ( P ＜ 0.05 ),while the difference between one year after therapy and two years after therapy was not statistically significance ( P ＞ 0.05 ).The first and second year recurrence rate was 2％ and 6％ in the DC group,while in the control group it was 20％ and 30％.The difference was statistically significant ( P ＜ 0.05 ).Conclusion Second transurethral resection combined with instillation therapy and transfusion therapy of dendritic cells pulsed with tumor cells could be an effective therapeutic approach to lower the recurrence rate on non muscle-invasive bladder cancer.

OBJECTIVETo identify genes associated with metastasis suppression and to investigate the molecular mechanism of osteosarcoma metastasis.

METHODSThe subtracted cDNA library of low metastatic human osteosarcoma cell line SOSP-9607 was constructed using suppression subtractive hybridization. Partial clones were sequenced. The acquired sequence data were aligned against the GenBank nucleotide database using Blastn to search for sequence matches. The interested clone was used to perform Northern blot and reverse transcriptase-PCR (RT-PCR) analysis on mRNA isolated from low metastatic cell line SOSP-9607 and OS-9901, high metastatic cell line SOSP-M and three pulmonic metastatic nodules of nude mice.

RESULTSA cDNA clone from low metastatic cell line SOSP-9607 subtracted cDNA library was identified as telomeric repeat binding factor 2 (TERF2) by sequence analysis and Blastn search. Northern blot and RT-PCR analysis demonstrated that TERF2 expressed highly in low metastatic cell line SOSP-9607 and OS-9901, but not in high metastatic cell line SOSP-M and three pulmonic metastatic nodules.

CONCLUSIONTERF2 may be important for suppressing metastasis of osteosarcoma.

Animals ; Base Sequence ; Blotting, Northern ; DNA-Binding Proteins ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Mice, Nude ; Molecular Sequence Data ; Neoplasm Metastasis ; genetics ; Neoplasm Transplantation ; Nucleic Acid Hybridization ; methods ; Osteosarcoma ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Telomeric Repeat Binding Protein 2 ; Transplantation, Heterologous ; Tumor Cells, Cultured

Background: and Purpose Brainstem gliomas (BSGs) in adults are rare brain tumors with dismal outcomes. The aim of this study was to determine the clinical and genetic features in a series of BSGs and their association with the prognosis. Methods: Fifty patients who underwent a stereotactic biopsy between January 2016 and April 2018 at a single institution were collected. Data on clinicopathological characteristics were analyzed and factors associated with patient survival were identified using a Cox regression model. Results: The median age at diagnosis was 55.5 years, and 62% of the patients were male. Glioblastoma (44%) accounted for the largest proportion of BSGs, and oligodendroglioma (2 of 50) was rarely encountered. The IDH mutation (6 of 44) occurred infrequently in astrocytomas, and IDH-mutant tumors harbored both ATRX loss and MGMT promoter methylation at a relatively low level. Wild-type IDH astrocytomas were identified as having high rates of 1p/19q codeletion (5 of 38) and loss of heterozygosity 1p (8 of 38) or 19q (8 of 38) only. In diffuse midline glioma H3K27M mutant, MGMT promoter methylation occurred in three of four cases. Patients were offered radiotherapy and/or concurrent/adjuvant temozolomide chemotherapy, and their median survival time was 13 months. Multivariate analysis revealed that a low tumor grade, absence of tumor enhancement, duration of symptoms ≥3 months, Karnofsky performance status ≥70, and ATRX loss conferred a survival advantage. Conclusions Adult BSGs showed different molecular genetic characteristics, but also resembled supratentorial gliomas in their clinical features associated with oncological outcomes.

Background: and Purpose Brainstem gliomas (BSGs) in adults are rare brain tumors with dismal outcomes. The aim of this study was to determine the clinical and genetic features in a series of BSGs and their association with the prognosis. Methods: Fifty patients who underwent a stereotactic biopsy between January 2016 and April 2018 at a single institution were collected. Data on clinicopathological characteristics were analyzed and factors associated with patient survival were identified using a Cox regression model. Results: The median age at diagnosis was 55.5 years, and 62% of the patients were male. Glioblastoma (44%) accounted for the largest proportion of BSGs, and oligodendroglioma (2 of 50) was rarely encountered. The IDH mutation (6 of 44) occurred infrequently in astrocytomas, and IDH-mutant tumors harbored both ATRX loss and MGMT promoter methylation at a relatively low level. Wild-type IDH astrocytomas were identified as having high rates of 1p/19q codeletion (5 of 38) and loss of heterozygosity 1p (8 of 38) or 19q (8 of 38) only. In diffuse midline glioma H3K27M mutant, MGMT promoter methylation occurred in three of four cases. Patients were offered radiotherapy and/or concurrent/adjuvant temozolomide chemotherapy, and their median survival time was 13 months. Multivariate analysis revealed that a low tumor grade, absence of tumor enhancement, duration of symptoms ≥3 months, Karnofsky performance status ≥70, and ATRX loss conferred a survival advantage. Conclusions Adult BSGs showed different molecular genetic characteristics, but also resembled supratentorial gliomas in their clinical features associated with oncological outcomes.

Objective To analyze the relevant factors that influence the medicine-free re-mission rate after short-term continuous subcutaneous insulin infusion (CSII)in newly diagnosed patients with type 2 diabetes metillus (T2DM).Methods A total of 405 patients with T2DM hos-pitalized in our hospital from October 2003 to October 2012 served as the study objects,who were given CSII for 15 d and divided into success group (165 cases)and failure group (240 cases)based on the requirement of hypoglycemic agents after discharge from hospital.The relationship between ages,genders,body max index (BMI),fasting plasma glucose (FPG),glycated hemoglobin (HbA1c),ratio of 2 h C-peptide and fasting peptide (C2 /C0),attained time of plasma glucose and daily dosage of insulin during CSII with the clinical medicine-free remission rate after short-term CSII in patients with T2MD.Results Success group was evidently lower in FPG,HbA1c and daily dosage of insulin but obviously higher in C2 /C0 during CSII than failure group,and the differences were all statistically significant (P <0.01).Conclusion FPG,HbA1c and C2 /C0 can be used as the screening indexes for the diagnosis of T2DM patients treated with CSII,and T2DM patients with low FPG and HbA1c and high C2 /C0 are more suitable and should given CSII earlier so as to induce longer-term remission rate in clinic.