Objective To study the effect of dualdi butyryl cyclic AMP (dbcAMP)on the proliferation of FTC-133 cell line. Methods FTC-133 cells were normally cultured and divided into control group,dbcAMP treatment group (0.5,1.0,2.0 mmol/L). After FTC-133 cells were treated with dbcAMP (0.5,1.0,2.0 mmol/L)for 24 h or 48 h,the growth activity and growth curve was detected by MTT.Changes of the cell cycle were detected by flow cytometry.The mRNA and protein expression of Raf1 were measured by RT-qPCR and Western blotting.Results Compared with control group,the growth activity of FTC-133 cells was re-duced by different levels of dbcAMP in a dose-time dependence manner.The number of FTC-133 cells was decreased in the S phase and increased in the G2/M phase.The mRNA and protein expression of Raf1 of treatment group were both reduced compared with control group.Conclusion dbcAMP significantly reduced FTC-133 cells proliferation and promoted apoptosis,and which might be involoved by ERK MAPK signalling.

This study is to investigate the mechanism and action characteristics of 6-chloro-3-methyl-4-(2-methyoxycarbonylthiophene-3-sulfonyl)-3, 4-dihydroquinoxa-lin-2-(1 H)-one (XU07011) against HIV-1 replication. XU07011 anti-HIV activity was tested by using VSVG/HIV pseudotype viral system and confirmed by HIV-1 live viruses' infectious assay. Time of addition was used to test HIV-1 reverse transcription process. RNA-dependent DNA polymerase activity and RNase H activity were tested by using enzyme linked immunoabsorbent assay and fluorescence method. Wild type and nine NNRTIs-resistant reverse transcriptase enzymatic models and cell-based pharmacological models were used to evaluate XU07011 bio-characteristics. The results showed that XU07011 inhibited HIV-1 replication with IC50 of (0.057 +/- 0.01) micromol x L(-1) which was comparable to nevirapine [IC50: (0.046 +/- 0.01) micromol x L(-1)]. Mechanism study data indicated that XU07011 blocked HIV-1 reverse transcription process through acting on reverse transcriptase RNA-dependent DNA polymerase with IC 50 of (1.1 +/- 0.3) micromol x L(-1). The compound showed no effect on RNase H activity. XU07011 exhibited better activities comparing with nevirapine on K103N mutated NNRTIs-resistant HIV-1 strains. This study could provide a theoretical basis for novel anti-HIV reagents development.

Background and purpose: For neoplasms with different sources, proteasome inhibitors can inhibit their growth and promote the cell apoptosis. Its mechanism may be associated with Bcl-2-associated athanogene 2 (BAG2). We aimed to investigate the involvement of in thyroid cancer cell death induced by proteasome inhibitors. Methods:A panel of thyroid cancer cells (ARO, FRO, KTC1, KTC2, KTC3, 8305C and 8505C) were treated with vehicle or proteasome inhibitor MG132;BAG2 mRNA and protein levels were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Results:MTT results indicated that FRO and KTC2 cell lines were the most sensitive to proteasome inhibitors. MG132 induced BAG2 mRNA and protein expression in the panel of thyroid cancer cells with various degree (P<0.01), for FRO and KTC2 cell line, the BAG2 mRNA level was increased in 20 to 25 times, meanwhile the protein expression level also increased signiifcantly;Sensitive FRO cells demonstrated quicker induction of BAG2 compared with insensitive ARO cells. Moreover, the extents of BAG2 induction in FRO cells were higher than that in ARO cells. Conclusion: BAG2 is a novel molecule induced by proteasome inhibitor, which might function as a proapoptotic molecule in thyroid cancer cell death mediated by proteasome inhibitor.

Objective To investigate the clinical manifestations, histopathological patterns, and methods of diagnosis and treatment of primary malignant duodenal tumors. Methods The data of 82 patients with primary malignant duodenal tumors confirmed by pathology and admitted to our hospital over a 10-year period were analyzed retrospectively. Results Tumor location:Tumors were located in the peripapilla region in 64 cases, at the descending portion of the duodenum in 11 cases, at other regions of the duodenum in 7 cases. The common presenting symptoms and signs were abdominal pain in 57 cases, jaundice 53 cases, and gastrointestinal bleeding in 41 cases. In these patients, radical resection of tumor was performed in 36 cases, palliative resection of tumor in 31cases,and operative intervention was not done in 15 cases.The 5-year survival rate of followed-up patients in this group was 2.4%. Conclusions The common presenting symptoms and signs of patients with primary malignant duodenal tumors were abdominal pain, jaundice and GI bleeding, but these patients usually lack specific symptoms and signs. The chief pathologic type is adenocarcinoma and the predisposed site of occurrence is the duodenal papillary region and the descending duodenum . CT, B ultrasonography and gastroduodenoscopy are the chief measures for the diagnosis of primary duodenal malignant tumors, and surgical resection is the main modality of treatment of this disease. The prognosis of primary duodenal malignant tumors is very poor.

Objective:To determine the clinicopathological significance of the DNA methyltransferase 3B (DNMT3B)overexpression in endometrial carcinomas and to evaluate its correlation with hormone re-ceptor status.Methods:Immunohistochemistry was performed to assess the expression of DNMT3B and hormone receptors in 104 endometrial carcinomas.Results:DNMT3B overexpression occurred frequently in endometrioid carcinoma (EC,54.8%)more than in nonendometrioid carcinoma (NEC,30.0%) with statistical significance (P =0.028).Furthermore,there was a trend that EC with worse clinico-pathological variables and shorter survival had a higher DNMT3B expression,and the correlation between DNMT3B and tumor grade reached statistical significance (P =0.019).A negative correlation between DNMT3B and estrogen receptor (ER)or progesterone receptor (PR)expression was found in EC. NMT3B overexpression occurred frequently in the ER or PR negative subgroups (78.9%,86.7%)more than in the positive subgroups (47.7%,47.8%)with statistical significance (P =0.016,P =0.006). In addition,the DNMT3B overexpression increased in tumors with both ER and PR negative expression (92.9%,P =0.002).However,no such correlation was found in NEC (P >0.05).Sequence analyses demonstrated multiple ER and PR binding sites in the promoter regions of DNMT3B gene.Conclusion:This study showed that the expression of DNMT3B in EC and NEC was different.DNMT3B overexpres-sion in EC was associated with the worse clinicopathological variables and might have predictive value. The methylation status of EC and NEC maybe different.In addition,in EC,DNMT3B overexpression negatively correlated with ER or PR expression.In NEC,the correlation between DNMT3B and ER or PR status was not present.

Objective To investigate the mechanism responsible for the malignant progressionof meningiomas at the protein level using tissue microarray technique. Methods Twenty-twointracranial meningioma tissue microarrays were constructed, each containing the tissues of 42 benign, 18atypical, and 19 anaplastic meningiomas. Immunohistochcmistry of the microarrays was performed induplicate with the antibodies of MYC, ARNT2, MDM2, AR, ER, PR, Ki-67, P53, survivin, CD34 andVEGF, respectively. Negative control microarrays were used throughout the experiment and breast cancertissue microarrays were used as the positive controls for ER and PR staining. SAS9.0 solfware was usedfor grading of the expression levels of the biomarkers according to the WHO grades of meningiomas.Results For each antibody, the duplicate tissue microarrays yielded uniform staining results invisualization of the protein distributions in the cytoplasm and nuclei, and the negative controls displayedno positive staining. The p53, AR, ER, PR and Ki-67 proteins were found only in the cell nuclei, MDM2in both the cytoplasm and nuclei, and ARNT2, CD34 and VEGF in the cytoplasm only. The c-MYC andsurvivin proteins were found mainly in the cytoplasm, and in some instances in both the cytoplasm andcell nuclei. Immunohistochemical staining for p53, AR, CD34, Ki-67 and MYC proteins showed strongcorrelations to the degree of malignancy of the meningioma (P<0.05). Conclusions Tissue microarrayand immunohistochemical techniques provide an efficient means for screening the specific biomatkers ofmeningiomas. The expressions of p53, AR, CD34, Ki-67 and MYC proteins are involved in the malignantprogression of meningioma, and these proteins may serve as important biomarkers for meningiomagrading at the protein level.

Lynch syndrome is an autosomal dominant genetic disease characterized by the early onset of colon cancer, endometrial cancer and other tumors caused by a genetic mutation within DNA mismatch repair (MMR) genes.A small subgroup (approximately 3% -5%) of endometrial cancer and colorectal cancer is related to Lynch syndrome .Identification of these patients in clinical practice will be of great benefit to the relatives and patients themselves .We reported two cases, and reviewed the literature and clinical diagnostic guideline.MMR protein was lost in the tumors.Meanwhile the two cases had different clinicopathological characteristics.Together with the literature, our findings may suggest that the MMR protein expression, associated molecular alterations and clinicopathological features and biological behavior of endometrial cancer and colorectal cancer related to Lynch syndrome are different .Thus the algorithm for detection the patients at highest risk is different .To detect the MMR loss by immunohisto-chemistry is a practicalscreening method.

BACKGROUND:Human mesenchymal stem cells derived from Wharton's jelly(WJCs)display the characteristics of MSCs as defined by the International Society for Cellular Therapy.They can be differentiated into bone,cartilage,adipose,muscle,and neural cells.They can also support the expansion of other stem cells,be weli-tolerated by the immune system,and have the ability to home to tumors.OBJECTIVE:To investigate biological changes of WJCs and brain tumor stem cells(BTSCs)co-cultured in vitro.METHODS:WJCs cultured by situ cultivation and BTSCs used enzyme digestion way respectively,and gathering the 3rd passage of WJCs though subculturing as well as BTSCs.Two kinds of cells co-cultured in 24-well plates in serum-free medium (SFM)without any growth factor.3 and 7 days after co-cultured respectively,CD133 expression of suspension cells in the 24-well plates were identified by flow cytometry,and immunofluorescence was performed for Nestin and glial fibrillary acidic protein (GFAP)expression of adherent cells.Co-culture supernatant(CCS)re-suspended 3~(rd) passage of BTSCs and cultured into 96-well plates at day 3,which were used to determine the difference in cell growth curve in both groups using a microplate reader.RESULTS AND CONCLUSION:With the cocultivation days increasing,the phenomenon that tumor sphere cells began to be decomposed,adherent and differentiated observed by an inverted microscope.BTSCs in the co-cultured group expressed GFAP and Nestin when adherent and differentiated.The higher degree of malignant brain tumor tissue used in culturing BTSCs was,the higher expression of CD133 in BTSCs was.CD 133~+ in BTSCs declined when co-cultured with WJCs.Growth curve of brain tumor stem cells cultured in CCS compared with in SFM at day 3,which indicates that the proliferation of BTSCs inhibited obviously.Results indicated that CD 133~+ expression and proliferative capacity of BTSCs went down and BTSCs underwent differentiation during the co-culture in vitro.

Objective: To observe the effects of acupoint thread-embedding therapy and low-carbohydrate diet therapy on obese patients with food addiction. Methods: Sixty-five eligible patients were randomized into a thread-embedding group of 33 cases and a diet group of 32 cases to respectively receive 12-week treatment. Before treatment, after treatment and at 6-month follow-up, the two groups were observed and compared in terms of body mass (BM), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body mass index (BMI), body fat rate (BFR), basal metabolic rate (BMR) and Yale food addiction scale version 2.0 (YFAS 2.0). Results: At the end of treatment, there were no significant differences in the general efficacy, and the improvements in BM, BMI, WC, HC, WHR and BFR between the thread-embedding group and diet group (all P>0.05). At follow-up, the thread-embedding group showed more significant improvements in all the aforementioned indicators compared with the diet group except HC (all P<0.05). At the end of treatment and follow-up, BMR and YFSA 2.0 had more significant improvements in the thread-embedding group than in the diet group (all P<0.05). Conclusion: Acupoint thread-embedding therapy can produce significant efficacy in treating obese patients with food addiction; it can improve the food addiction state and work better in maintaining the efficacy compared with low-carbohydrate diet therapy.

Objective To study the expressions of poly(ADP-ribose) polymerase (PARP) and apoptosis-inducing factor (AIF) in the hippocampal CA1 region of rats after focal cerebral ischemia reperfusion injury,and elucidate the neuroprotective effect ofcilostazol pretreatment through PARP/AIF pathway.Methods One hundred and thirty-five male SD rats were randomly divided into three groups (n=45) for experiments:sham-operated group,ischemia/reperfusion (I/R) vehicle group and cilostazol pretreatment group.Embolization thread was inserted into all rats except those in the sham-operated group for 2 hours to establish middle cerebral artery occlusion models; and then,reperfusion for 6,24,72 hours was performed,and rats of each group were sub-divided into three groups according to reperfusion times (n=l 5).Cilostazol pretreatment group was given cilostazol (30 mg/kg,twice orally) before surgery.The apoptosis cells of the hippocampus were detected by Terminal deoxynucleotidyl Transferasemediated dUTP nick end-labeling (TUNEL).Western blotting was used to test the dynamic changes of AIF and polyadenosine diphosphate ribose (PAR) levels at different times.The mRNA expression of AIF was assessed by real time-PCR.Results There appeared subsequent translocation of AIF after focal cerebral ischemia reperfusion injury in rats.As compared with those in the sham-operated group,the apoptotic cells statistically increased; AIF and PAR contents and AIF mRNA expression significantly increased in the I/R vehicle group reaching its peak level at 24 h ofreperfusion (P＜0.05).As compared with of the I/R model subgroups,the cilostazol pretreatment subgroups had significantly decreased AIF and PAR contents,AIF mRNA expression and number of apoptosis cells (P＜0.05); 24 h of reperfusion subgroup had the most obvious effect (P＜0.05).Conclusion Cilostazol pretreatment in rats after ischemia-reperfusion injury has certain neroprotective effect,whose resistance roles in nerve cell apoptosis may be implemented by inhibiting excessive PARP activation and AIF translocation caused by cerebral ischemia injury.