Objective To study the apoptosis of ovarian carcinoma cell strain COC1 induced by arsenic trioxide (As_2O_3). Methods The effect of arsenic trioxide on the proliferation of ovarian carcinoma cell strains were examined, using Methyl thiazolyl tetrazolium(MTT) assay. Flow cytometry(FCM) was used to detect apoptosis percentage and phase distribution of cell cycles. After being exposed to As_2O_3 solution of different concentration, apoptosis morphologic features of COC1 were observed by TdT-mediated dUTP nick end labeling (TUNEL). Results Inhibition of As_2O_3 on the growth of COC1 was increasing with passing of time and increasing of concentration. After 48h of exposure to 1.5 ?mol/L As_2O_3, COC1 cell strains presented apoptosis morphologic change, and the number of apoptosis cells increased with the passage of time. After treatment with 3.0 ?mol/L or 1.5 ?mol/L As_2O_3, the percentage of COC1 cells in G_2/M phase declined, and the percentage of cells in G_1 phase increased with the passage of time. These results suggested that As_2O_3 inhibited the cellular proliferation of COC1 cells via arrest of cell cycle. Conclusion Arsenic trioxide can induce apoptosis of ovarian carcinoma cell strain COC1. It can block the cell cycle at the G_1 phase, which is one of the possible mechanisms of apoptosis induced by As_2O_3.

Objective To evaluate the predictive value of N-terminal-pro-brain natriuretic peptide (NT-proBNP) in weaning patients from mechanical ventilation (MV).Methods Data of 42 patients supported with MV in intensive care unit (ICU) admitted to the Rui Jin Hospital from January through December in 2014 were retrospectively analyzed,and the causes for MV were recorded.According to the outcomes of weaning from MV after 48 hours,the patients were divided into two groups namely success group and failure group.Comparisons of fluid balance in 72 hours before spontaneous breathing trial (SBT),and comparisons of NT-proBNP1 levels at admission,NT-proBNP2 levels before SBT,NT-proBNP3 levels after 48 hours after SBT between two groups were carried out.And the receiver operating characteristic (ROC) curve for predicting weaning rate was plotted to find the optimal cut-off point of NT-proBNP2.Results In the total of 42 patients,there were 27 cases in success group and 15 cases in failure group.There were not statistically differences of NT-proBNP1 levels between success group and failure group (P =0.121).However,the NT-proBNP2 levels and NT-proBNP3 levels in failure group were significantly higher than those in success group (P =0.01,0.003).The area under curve (AUC) of the ROC curve of NT-proBNP2 levels to predict the failure of weaning was 0.862 (95％ CI:0.753-0.971).When the optimal cut-off point of NT-proBNP2 was 715.5 pg/mL,the sensitivity and specificity were 93.3％ and 74.1％,respectively.Conclusion The NT-proBNP2 levels before SBT have predictive value in weaning rate,and it can be used as one of the screening indicators for weaning.

Objective To explore the mechanisms and effects of adoptive immunotherapy with ovarian cancer vaccine modified by GM-CSF gene which was used after immunologic reconstitution during lymphopenia induced by chemotherapy.Methods Lymphopenia was induced by chemotherapy with cyclophosphamide.The immune reconstituted model was built in rats.The tumor vaccine draining lymph nodes were harvested after the ovarian cancer cells NUTU-19 modified by GM-CSF gene were injected.The effector T cells (T_E) were got after being stimulated and amplified.Enzyme-linked immunosorbent assay was used to detect the level of interleukin (IL)-2 and IL-4 secreted by T_E.Intracellular cytokine staining was used to determine frequency of tumor-specific T_E.Fluorescence-activated cell sorting (FACS) was used to detect the special cytotoxicity ofT_E killing target cells.The survival period of rats bearing pre-established abdominal ovariam carcinoma after being adoptively transferred byT_E.was observed.Results Compared with those in control group,the significant higher levels IL-2[(65.7±4.0) pg/ml]and lower levels IL-4 [(277±49) pg/ml]were observed in chemotherapy-immune recunstitution-vaccine immunization group.The amount of CD_4~+ T cells secreting interferon-γ (13.0±2.1)% were also significantly increased.The rate of the special cytotoxicity of killing T cells (86.5±1.1) % was markedly improved.The survival period of rats (110±16) days was increased in chemotherapy-immune reconstitution-vaccine immunization group.Conclusions The combined immunotherapy of chemotherapy-immune reconstitution-tumor vaccine immunotherapy may increase the frequency and function of specific tumor T_E.The specific cytotoxicity is increased and the weak reaction of T_E to tumor is improved,which showed that this therapy can enhance immune reaction.

OBJECTIVETo investigate and establish the gene diagnosis methods for the frequent mutations of iduronate-2-sulphatase(IDS) gene in mucopolysaccharidosis type II patients.

METHODSpolymerase chain- reaction-single strand conformation polymorphism PCR-SSCP) analysis was applied to detect the mutations of exons 3, 8 and 9 which were hot spots in the iduronate-2-sulfatase gene; DNA sequencing was applied to analyze the mutations which had been detected by PCR-SSCP; PCR-restriction fragment length polymorphism (PCR-RFLP) was applied to detect the results of DNA sequencing.

RESULTSObvious and abnormal bands in exon 9 of the IDS gene were found by applying PCR-SSCP; the mutation(C1672T) of exon 9 was found in the patient through DNA sequencing, which led to amino acid replacement(R468W); the PCR-restriction enzyme digestion showed that only one band(554 bp) appeared in the patient, but there were two bands (257 bp and 297 bp) in his parents, and it verified the results of sequencing analysis.

CONCLUSIONPCR-SSCP analysis, DNA sequencing analysis and PCR-restriction enzyme digestion are effective methods for MPS II diagnosis. Combined applications of these methods can verify and complement each other and improve the accuracy of diagnosis.

Amino Acid Substitution ; Base Sequence ; Child ; China ; Codon, Nonsense ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Mucopolysaccharidosis II ; enzymology ; genetics ; Mutation ; Point Mutation ; Polymorphism, Single-Stranded Conformational

Chromosomal region maintenance 1 (CRM1) is associated with an adverse prognosis in glioma. We previously reported that CRM1 inhibition suppressed glioma cell proliferation both in vitro and in vivo. In this study, we investigated the role of CRM1 in the migration and invasion of glioma cells. S109, a novel reversible selective inhibitor of CRM1, was used to treat Human glioma U87 and U251 cells. Cell migration and invasion were evaluated by wound-healing and transwell invasion assays. The results showed that S109 significantly inhibited the migration and invasion of U87 and U251 cells. However, mutation of Cys528 in CRM1 abolished the inhibitory activity of S109 in glioma cells. Furthermore, we found that S109 treatment decreased the expression level and activity of MMP2 and reduced the level of phosphorylated STAT3 but not total STAT3. Therefore, the inhibition of migration and invasion induced by S109 may be associated with the downregulation of MMP2 activity and expression, and inactivation of the STAT3 signaling pathway. These results support our previous conclusion that inhibition of CRM1 is an attractive strategy for the treatment of glioma.

Chromosomal region maintenance 1 (CRM1) is associated with an adverse prognosis in glioma. We previously reported that CRM1 inhibition suppressed glioma cell proliferation both in vitro and in vivo. In this study, we investigated the role of CRM1 in the migration and invasion of glioma cells. S109, a novel reversible selective inhibitor of CRM1, was used to treat Human glioma U87 and U251 cells. Cell migration and invasion were evaluated by wound-healing and transwell invasion assays. The results showed that S109 significantly inhibited the migration and invasion of U87 and U251 cells. However, mutation of Cys528 in CRM1 abolished the inhibitory activity of S109 in glioma cells. Furthermore, we found that S109 treatment decreased the expression level and activity of MMP2 and reduced the level of phosphorylated STAT3 but not total STAT3. Therefore, the inhibition of migration and invasion induced by S109 may be associated with the downregulation of MMP2 activity and expression, and inactivation of the STAT3 signaling pathway. These results support our previous conclusion that inhibition of CRM1 is an attractive strategy for the treatment of glioma.

OBJECTIVETo clone a novel gene which is related to human testis spermatogenesis apoptosis.

METHODSTo rapidly attain human novel gene full-length cDNA sequence from a human testis cDNA library,the gene-specific primers and the vector-specific primers were designed for nested polymerase chain reaction. Sequencing was performed and the result was analysed.

RESULTSThe present authors discovered the TSARG3 gene(GenBank accession number AF419291) from a human testis cDNA library, using a cDNA fragment (GenBank accession number BE644537) as an electronic probe, which was significantly changed in cryptorchidism and represented a novel gene. Furthermore, a mouse homologue of this gene was identified (GenBank accession number AF419292) by using the same method.

CONCLUSIONA novel gene named TSARG3 was cloned. It is considered that the function of the new gene is related to human testis spermatogenesis apoptosis.

Amino Acid Sequence ; Animals ; Apoptosis ; genetics ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Female ; Gene Expression ; Heat-Shock Proteins ; Humans ; Male ; Mice ; Molecular Sequence Data ; Proteins ; genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Spermatocytes ; cytology ; metabolism

Burning mouth syndrome (BMS) is a complex chronic disorder characterized symptomatically by burning sensation mainly localized in the tongue with no specific lesions. Its etiology is not fully understood. Due to the persistent pain and unsatisfactory outcome of treatment, patients with high expectation when seeking medical advice easily feel discontent after treatment, which might cause medical dispute. This review introduces the epidemiology, manifestations, etiology, diagnosis, classification and treatment of BMS. The authors hope the current advances about BMS, mainly focus on the etiology of biological nerve and social psychology, may help the clinicians in treatment of BMS.

BACKGROUND: Diabetic wound healing remains a major challenge due to the impaired functionality of angiogenesis by persistent hyperglycemia. Mesenchymal stem cell exosomes are appropriate candidates for regulating the formation of angiogenesis in tissue repair and regeneration. Here, we explored the effects of exosomes derived from human amniotic mesenchymal stem cell (hAMSC-Exos) on the biological activities of human umbilical vein endothelial cells (HUVECs) treated with high glucose and on diabetic wound healing and investigate lncRNAs related to angiogenesis in hAMSC-Exos. METHODS: hAMSCs and hAMSC-Exos were isolated and identified by flow cytometry or western blot. A series of functional assays such as cell counting kit-8, scratching, transwell and tube formation assays were performed to evaluate the potential effect of hAMSC-Exos on high glucose-treated HUVECs. The effect of hAMSC-Exos on diabetic wound healing were tested by measuring wound closure rates and immunohistochemical staining of CD31. Subsequently, the lncRNAs profiles in hAMSC-Exos and hAMSCs were examined to screen the lncRNAs related to angiogenesis. RESULTS: The isolated hAMSC-Exos had a size range of 30–150 nm and were positive for CD9, CD63 and CD81. The hAMSC-Exos facilitate the functional properties of high glucose-treated HUVECs including the proliferation, migration and the angiogenic activities as well as wound closure and angiogenesis in diabetic wound. hAMSC-Exos were enriched lncRNAs that related to angiogenesis, including PANTR1, H19, OIP5-AS1 and NR2F1-AS1. CONCLUSION Our findings demonstrated hAMSC-Exos facilitate diabetic wound healing by angiogenesis and contain several exosomal lncRNAs related to angiogenesis, which may represent a promising strategy for diabetic wound healing.