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.

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.

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.

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.

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.

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.

A total of 169 patients with short-duration type 2 diabetic mellitus (DM) were divided into atherosclerosis (AS) group and non-AS group according to the intima-media thickness (IMT) of three conducting arteries.The level of serum amyloid A (SAA) was assayed by ELISA.The results showed that SAA level of type 2 DM patients increased significantly, patients in AS group showed higher SAA level than that in non-AS group, and SAA level was positively correlated with age, body mass index, waist hip ratio and IMT of common carotid artery.Age, C-reactive protein and SAA level are the major risk factors for IMT of common carotid artery.

OBJECTIVETo discuss the protective effect of alkaloids from Piper longum (PLA) in rat dopaminergic neuron injury of 6-OHDA-induced Parkinson's disease and its possible mechanism.

METHODThe rat PD model was established by injecting 6-OHDA into the unilateral striatum with a brain solid positioner. The PD rats were divided into the PLA group (50 mg x kg(-1) x d(-1)), the madorpa group (50 mg x kg(-1) x d(-1)) and the model group, with 15 rats in each group. All of the rats were orally given drugs once a day for 6 weeks. Meanwhile, other 15 rats were randomly selected as the sham operation group, and only injected with normal saline in the unilateral striatum. The behavioral changes were observed with the apomorphine (APO)-induced rotation and rotary rod tests. The number of tyrosine hydroxylase (TH)-positive cells in rat substantia nigra and the density of TH-positive fibers in striatum were detected by tyrosine hydroxylase immunohistochemistry. The content of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH), catalase (CAT), malondialdehyde (MDA), nitric oxide (NO) and nitric oxide synthase (NOS) in rat substantia nigra and striatum were measured by the spectrophotometric method.

RESULTAfter being induced by APO, PD rats showed obvious rotation behaviors, with decreased time stay on rotary rod and significant reduction in the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. The activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity significantly decreased, whereas the activities of NOS and the content of MDA, NO significantly increased. PLA could significantly improve the behavioral abnormality of PD rats and increase the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. It could up-regulate the activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity, and decrease the content of NOS and the content of MDA, NO.

CONCLUSIONAlkaloids from P. longum shows the protective effect in substantia nigra cells of 6-OHDA-induced PD model rats. Its mechanism may be related with their antioxidant activity.

Administration, Oral ; Alkaloids ; administration & dosage ; pharmacology ; Animals ; Apomorphine ; pharmacology ; Catalase ; metabolism ; Dopamine Agonists ; pharmacology ; Dopaminergic Neurons ; drug effects ; metabolism ; pathology ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Motor Activity ; drug effects ; Neostriatum ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; physiopathology ; prevention & control ; Piper ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; drug effects ; metabolism ; Superoxide Dismutase ; metabolism ; Tyrosine 3-Monooxygenase ; metabolism

OBJECTIVETo study toxic effects of 2,5-hexanedione (2,5-HD) on pathology and lipid peroxidation in mouse retina.

METHODSForty-eight mice were randomly divided into blank control group (12 mice), negative control group exposed to normal solution (12 mice) and group exposed to 2,5-HD for 2. 4 and 8 weeks, respectively (24 mice) by intraperitoneal injection (2.5% 2,5-HD) at the dose of 400 mg/kg. The pathological changes of mouse retina were examined under light microscope. The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in mouse retina were detected.

RESULTSThe retinal structure in the blank and negative control groups was normal. In mice exposed to 2,5-HD for 8 weeks, the swelling of outer and inner segments and disorder arrangement of the segments without clear boundary were found. The staining of outer plexiform layers was uneven and the irregular loose structure appeared. The hyperchromatic pyknotic and necrosis nuclei were presented in ganglion cells layer. Compared with the control and blank groups, the activities of SOD gradually and significantly reduced and the concentrations of MDA increased in group exposed to 2,5-HD (P < 0.05).

CONCLUSION2,5-HD can induce the injury of retina tissues of mice, which may be associated with the lipid peroxidation.

Animals ; Hexanones ; toxicity ; Lipid Peroxidation ; drug effects ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred Strains ; Retina ; drug effects ; metabolism ; pathology ; Superoxide Dismutase ; metabolism