OBJECTIVETo study the expression and associations of three survivin splicing variants in gastric cancer and normal gastric mucosa, and to evaluate the prognostic significance.

METHODSReal time quantitative RT-PCR was used to detect the expression of three survivin splicing variants in tumor and matched normal gastric mucosa specimens from 77 cases with gastric cancer.

RESULTSThe expression of three survivin splicing variants than upregulated significantly in gastric cancer than those in normal mucosas (P< 0.01). In cancer tissues, the expression rates of survivin, survivin-2B, survivin-deltaEx3 were 100%, 79.8% (61/77), 64.9% (50/77) respectively. The survival rate was significantly lower in the patients with high survivin expression than those with low survivin expression (P< 0.01).

CONCLUSIONAmong three survivin splicing variants, the expression level of wild-type survivin mRNA is an important predictor for prognosis.

Adult ; Aged ; Aged, 80 and over ; Biomarkers ; Female ; Follow-Up Studies ; Gastric Mucosa ; pathology ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; genetics ; Neoplasm Staging ; Prognosis ; Protein Isoforms ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; genetics ; pathology ; Transcription, Genetic

Objective To choose proper proton magnetic resonance spectroscopy(~1XH-MRS) parameters to fit for practical femoral marrow cavity and to produce short-timed,well-repeated and excellent ~1H-MRS images.Methods The tentative study of ~1H-MRS on the normal femoral bone marrow in 26 volunteers was performed with a 1.5 T MR after the informed consent.The single-voxel spectroscopy and stimulated echo acquisition mode were used for ~1H-MRS collection.~1H-MRS parameters for 12 volunteers were 128 acquisitions,1 cm?1 cm?1 cm volume of interest(VOI)size and repeatedly 2—3 times within the same location.~1H-MRS parameters for another:14 volunteers were different numbers of acquisition (128 and 256 times,respectively)and different VOI sizes(2 cm?2 cm?2 cm and 1 cm?1 cm?1 cm, respectively).Results For ~1H-MRS with 1 cm?1 cm?1 cm size of VOI and 128 times of acquisition with the full width haft max of water≤8—12 Hz,the base-line was steady and the signal-noise ratio was high up to 11.31.~1H-MRS was different in the different femoral locations showing the maximum peak sites at near 0.90 ppm(?10~(-6))or 1.65 ppm,but~1H-MRS within the same location was always same or similar with different VOI sizes(1 cm?1 cm?1 cm or 2 cm?2 cm?2 cm)or different numbers of acquisition(128 or 256 times).~1H-MRS acquisition time was not related with the size of VOI but with the numbers of acquisition.128 and 256 times of acquisition cost 199 s and 391 s,respectively.Conclusion With the technique of small size of VOI(1 cm?1 cm?1 cm)and decreased numbers of acquisition(128 times),it is propable to get well-repeated and excellent ~1H-MRS within less time.It is also more practical for clinics to achieve ~1H-MRS of the femoral marrow with the proper technique.

The α4β2-nicotinic acetylcholine receptor (nAChR) is a ligand-gated ion channel that is distributed throughout the nervous system. It is involved in the regulation of various neurotransmitters including acetylcholine, dopamine, γ-aminobutyric acid, and norepinephrine. α4β2-nAChR plays an important role in learning, memory, cognition, attention, inflammation, and pain. A large number of studies have shown that α4β2-nAChR is an important therapeutic target for neurological diseases such as Alzheimer's disease, Parkinson's disease, epilepsy, depression, nicotine dependence, pain, etc. It is an important target in the early diagnosis and curative effect detection of neurodegenerative diseases including Alzheimer's disease. This review summarizes the role, mechanisms and related drug research advances on α4β2-nAChR ligand drugs in neurological diseases, as well as providing a theoretical basis for identifying and developing more suitable α4β2-nAChR-related compounds.

OBJECTIVETo investigate the effects and the mechanisms of cell growth inhibition in hepatocellular carcinoma cells after induction with antisense survivin-liposome (LIP) complex, and to provide evidence in treatment for hepatocellular carcinoma and tumors expressing survivin.

METHODSSurvivin ODNs was transfected into HepG2 cells mediated by LiP reagent. The expression of survivin mRNA and protein was detected by RT-PCR and Western blot. MTT assay was applied to determine cell proliferation in HepG2 cells. Active caspase-3 and apoptosis rate were evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Cell cycle was analyzed by flow cytometry in the cell cycle-synchronized hepatocellular carcinoma cells treated with the antisense compound.

RESULTSAntisense compound efficiently down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC(50) of 250 nmol/L. Its maximal effect was achieved at a concentration of 600 nmol/L, when expression levels were down-regulated by 80%, as revealed by gradually increase of caspase-3-like protease activity and apoptosis rate in a time-dependent manner. Morphological apoptotic changes such as membrane blebbing, loss of microvilli, cytoplasmic vasculization, condensation of cytoplasm and nucleus, chromatin fragmentation, and apoptosis and cell growth inhibition were observed. In the cell cycle-synchronized hepatocellular carcinoma cells, antisense compound induced cell cycle arrest followed by apoptosis. After treated with low concentration of compound, the cell cycle was arrested at S phase or G2/M phase; while at high concentration, the cell cycle was mainly arrested at S phase. Apoptosis was obviously observed and the rate of apoptosis was increased in a time and concentration-dependent manner.

CONCLUSIONAntisense survivin has significant inhibitory effect on growth of hepatocellular carcinoma cells in vitro. This is associated with cell cycle arrest and apoptosis.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; Liposomes ; Liver Neoplasms ; pathology ; Microtubule-Associated Proteins ; metabolism ; pharmacology ; Neoplasm Proteins ; metabolism ; pharmacology ; Oligonucleotides, Antisense ; pharmacology

Objective To investigate the children's body environmental Se and T-2 toxin level in their staple food in Kaschin-Beck disease(KBD)relative active regions in Aba state of Sichuan province in 2008.Methods We took X-ray photograph of the right hand on children aged 7-13 years in 48 villages from 11 counties in Aba state.The relative active regions of KBD were chosen according to the X-ray result and historical status of KBD.The children's urine and hair,drinking water and their staple food werr sampled.Selenium contents in urine,hair,water and food samples were determined by naphthalene fluorescence,and T-2 toxin in staple food samples were detected by ELISA kits.Results In 2145 X-ray films,66 films were positive,and the children's KBD positive rate was 3.08%(66/2145).The KBD positive rate was respectively 10.98%(29/264)and 8.52%(19/223)in Maerkang county,Jinchuan county and it was 0.75%(3/400)in Rangtang county,historically serious endemic area.The selenium content in urine of children aged 7-13 years in Maerkang county,Jinchuan county and Rangtang county was (10.41±4.67), (10.11±3.65), (8.42±2.68)μg/g Cr, respectively, there was no statistical difference among three counties(F=0.901, P>0.05). The selenium content in hair of children aged 7-13 years in Maerkang county[(0.18±0.04)mg/kg] was lower than that in Jinchuan county[(0.21±0.04)mg/kg, P<0.05].The selenium content in water in Jinchuan county [(0.225±0.124 )μg/L ] was lower than that in Maerkang county and Rangtang county[(0.320±0.092), (0.339±0.105)μg/L, all P<0.05]. The selenium content in staple food in Jinchuan county(0.0033 mg/kg) was lower than that in Maerkang county and Rangtang county(0.0258,0.0137mg/kg, Z=-6.146,-3.042, all P<0.017). The T-2 toxin level in flour in three counties was 19.60,17.95,26.25 ng/g,respectively,there was no statistical difference among three counties(X2=5.623, P>0.05).The T-2 toxin level in grain Maerkang county (10.72 ng/g) was higher than that in Jinchuan county and Rangtang county (3.74,3.30 ng/g, Z=-6.315,-4.407,all P<0.017). T-2 toxin contamination in flour was more severe than that in grain (Z=-6.690,-5.493,-3.676, all P<0.05). Conclusions In 3 relative active KBD regions of Aba state,the children's selenium nutritional status and the T-2 toxin contamination level in their staple food is consistent with the distribution of KBD.

Objective To investigate the expression of the interleukin-1 receptor(IL-1R)Ⅰ,IL-1RⅡand IL-1R accessory protein(IL-1RAcP)in osteoarthritis and analyse their biological significance.Methods Immunohistochemistry and reverse transcription-polymerase chain raction(RT-PCR)were adopted to detect the expression of IL-1RⅠ,IL-1RⅡand IL-1RAcP on the synovium of 107 OA patients.Results Immunohis- tochemistry showed strong positive expression of IL-1RⅠand IL-1RAcP,and positive expression of IL-1RⅡ. The expression was distributed in lining cells,monocyts and vascular endothelial cells of the sublining area, but all of them were negative or weak positive in normal synoviums.RT-PCR showed the expression of IL-1RⅠ,IL-1RⅡand IL-1RAcP in OA synoviums was significantly enhanced than normal synoviums (P＜0.05),and the expression of IL-1RⅠwas significantly enhanced than IL-1RⅡ(P＜0.05),but no sig- nificant difference with IL-1RAcP(P＞0.05).In stageⅡandⅢOA synoviums,the expression of IL-1RⅠand IL-1 RAcP had no significant difference with normal synoviums(P＞0.05).The expression of IL-1RⅡin stageⅢOA synoviums was significantly enhanced than normal(P＜0.05).Conclusion IL-1RⅠ,IL-1RⅡand IL-1RAcP play significant roles in the pathogenesis of OA,especially IL-1RⅠand IL-1RAcP.But their increase is only observed in the early stage of OA.These suggest that they may have no association with the development of OA and have no direct association with the severity of OA.OA can be cured by interrupting the signal transduction path in which IL-1 has played biological roles.

OBJECTIVE This study aimed to optimize polysaccharides extraction from Urena lobata L.and investigate its antioxidant activity.METHODS The mathematical model was established by re-sponse surface method (RSM) based on the results of single factor experiments, using polysaccha-rides extraction rate as response value,and using the ratio of water to material,cellulase concentra-tion,extraction temperature and time as experimental factors,which was used to screen optimum poly-saccharide extraction conditions from Urena lobata L.. Antioxidant activity of polysaccharides was stud-ied by DPPH and ·OH free radical elimination method. RESULTS The optimum conditions obtained by RSM were as follows:the cellulase level was 10.8 g·L-1,extraction time duration was 72 min,the ra-tio of water to feedstock was 7 mL·g-1,extraction temperature was 43℃,the pH value was 5.0.Under the optimal conditions, there was a difference of less than 5％ between predicted extraction rate 13.37％ and experimental extraction rate 13.32％. The polysaccharide yield was most significantly af-fected by cellulase concentration,followed by extraction time,water to material ratio and extraction tem-perature.IC50of DPPH and·OH were 1.082 g·L-1and 3.202 mg·L-1,respectively.Antioxidant activity of sample polysaccharides was weaker than those of vitamin C. CONCLUSION The polysaccharide extraction process from Urena lobata L. by cellulase enzymolysis approach was obtained, which was convenient and feasible,and extracted polysaccharides had good free radical scavenging activity.

Objective To explore the expression of hypoxia-inducible factor-1?(HIF-1?) protein and its mRNA in cultured cortical neurons after hypoxia,ischemia or hypoxia-ischemia(HI) and explore the possibilities of HIF-1? gene therapy in HI neurons.Methods The in vitro models of HI,pure hypoxia and pure ischemia were established using embryonic day 16-18 rats cortical neurons.Immunohistochemical and in-situ hybridization were performed to examine the expression of HIF-1? protein and its mRNA at different reperfusion time points in neurons.Results The expression of HIF-1? protein was very week in normoxic cultured neurons,but was up-regulated while treated with hypoxia and(or) ischemia.HIF-1? expression reached peak at 4 to 8 h after reperfusion with HI,which were statistically significant higher than that at other time points(Pa=0),and decreased gradually at 12 h.Furthermore,HIF-1? protein expression was significantly higher in HI group compared with that in the pure hypoxia or ischemia group(Pa=0).HIF-1? mRNA reached peak immediately after HI,decreased gradually at 2 h,and returned to the baseline at 8 h after reperfusion.Conclusions HIF-1? expression on cortical neurons is regulated differently with hypoxia,ischemia or HI treatment,HIF-1? gene therapy for HI neurons maybe a useful method in the future studies.

Objective To investigate the absorbability of the packaged infusion sets of butylphthalide,polyvinyl chloride (PVC) or polyolefin thermoplastic elastomer (TPE) infusion sets on butylphthalide for providing clinical reference in the choice of infusion sets.Methods Solution of butylphthalide was prepared as peractual concentration needed by the clinic,and dripped through PVC,TPE and packaged infusion sets respectively.The dripped solutions at different time points were collected and determined by HPLC,the contents were compared with that at 0 min.Results In the end of infusion,the adsorption rate of PVC,TPE and packaged infusion sets on butylphthalide were 0.47％,32.48％,9.62％,respectively.In addition,the absorbability of the infusion sets frown different manfacturers on butylphthalide had no significant differece.Conclusion The infusion sets of PVC had a serious absorption on butylphthalide.

OBJECTIVETo investigate the effects of protein tyrosine kinase (PTK), protein tyrosine phosphatase (PTP) and protein kinase C (PKC) on apoptosis and observe the changes of cytosolic calcium ([Ca(2+)]i) of arsenic trioxide (As2O3) treated human leukemia cells NB4 and cortex neurons.

METHODS[Ca(2+)]i of NB4 cells and cortex neurons was probed with Fluo-3/AM, its changes were assayed with laser confocal microscopy in real-time after As2O3 treatment at different concentrations, the effects of PTK and PTP and the activation of PKC on these changes with confocal microscopy and phosphorus radioisotope assay. DNA ladders of NB4 cells and cortex neurons after exposed to As2O3 were observed.

RESULTSAs2O3 at 1 micromol/L could remarkably increase the [Ca(2+)]i of NB4 cells but had no effects on neurons. Vanadate, a kind of PTP inhibitor, could promote the increase of [Ca(2+)]i treated by 2, 5, 10 micromol/L As2O3 in a dose-dependent manner. The mean total increase rates at 240 seconds after exposed to As2O3 at different concentrations were (6.5 +/- 2.3)%, (21.7 +/- 2.1)%, (49.2 +/- 2.5)% for NB4 cells, and (6.7 +/- 2.1)%, (19.4 +/- 2.5)%, (52.3 +/- 2.7)% for cortex neurons, respectively. Genistein, a kind of PTK inhibitor, could decrease the increase of [Ca(2+)]i treated by 2, 5, 10 micromol/L As2O3 in a dose-dependent manner. The mean total inhibited rates at 240 seconds after As2O3 treatment at different concentrations were (6.7 +/- 2.9)%, (25.6 +/- 2.5)%, (52.2 +/- 3.5)% for NB4 cells, and (7.8 +/- 3.1)%, (18.1 +/- 2.8)%, (51.3 +/- 3.3)% for cortex neurons, respectively. The activation of PKC began to increase as exposed to As2O3 at 1 micromol/L for 3 h, and kept rising continuously in NB4 cells and at 24 h DNA ladders emerged. However, none of the above results was found in human cortex neurons, but when exposed to 2 micromol/L As2O3, the activation of PKC and DNA ladders did emerge in neurons.

CONCLUSIONSThe phosphorylation and dephosphorylation of PTK and PTP participated in nonspecific apoptosis signal transduction pathway related to As2O3, and accompanied with PKC activation. The [Ca(2+)]i elevation was closely related to increased PKC activation. There existed difference in dose tolerances to As2O3 between NB4 cell and cortex neurons.

Adult ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Calcium ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Cerebral Cortex ; cytology ; Cytoplasm ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Leukemia, Promyelocytic, Acute ; enzymology ; metabolism ; pathology ; Male ; Neurons ; cytology ; drug effects ; metabolism ; Oxides ; pharmacology ; Protein Kinase C ; metabolism ; Protein Tyrosine Phosphatases ; metabolism ; Protein-Tyrosine Kinases ; metabolism