Objective To investigate the expressions of CYCLIND1 CDK4 in pancreatic cancer and their relationship with clinlicopathological parameters.Methods Immunohistochemical detector with new second generational tow steps method was used to detect the expressions of CYCLIND1 CDK4 and PCNA in 48 cases of pancreatic cancer and 14 cases of chronic pancreatitis.Results The expressions of CYCLIND1 CDK4 proteins in pancreatic cancer were 70.85%(34/48)and 62.50%(30/48) respectively.The CYCLIND1 and CDK4 proteins were increased(P

Objective To investigate the mechanisms of the inhibitory effects of peripheral electrical stimu- lation(PES)on chronic central pain(CCP)after spinal cord injury(SCI).Methods Twenty-four male Sprague- Dawley rats with CCP following SCI were randomly divided into three groups:a group without stainless steel needles implanted (NSSN group,n=8),a group with a stainless steel needle implanted but no peripheral electrical stimula- tion applied(NPES group,n=8)and a PES group(PES group,n=8).The rats' CCP was evaluated through ob- serving their response to nociceptive stimulation by means of the paw withdrawal pressure threshold(PWPT)and the paw withdrawal latency(PWL).Spontaneous pain behaviors including autophagia and scratching were observed at the same time.PES was applied via stainless steel needles inserted into standard acupoints on the hind limps and the back.The expression of the NMDA receptor 1(NR-1)subunit in the spinal cord horn was measured using immuno- chemical methods.Results Compared with the NSSN and NPES groups,CCP in the PES group was alleviated, PWPT and PWL were dramatically increased(P＜0.01)and the expression of NR-1 was obviously decreased (P＜0.01).Conclusion Peripheral electrical stimulation may alleviate chronic central pain after spinal cord injury in rats.

Objective:To value the application of two bacteria colony-PCR methods in the screening of phage antibody library. Methods:Five positive monoclonal bacterium were respectively suspended in either deionized water or 0.1% Triton X-100, and then boiled to be used as template in PCR. . The DNAs products of PCR were extracted and digested by two enzymes, and then determined by electrophoresis. Results:The inserted genes were detected in all the 5 clones after PCR and enzyme digestion .Conclusion:Bacteria colony-PCR can be used in screening positive recombinant colonies. The bacteria colony-PCR method with bacteria colonies suspended in deionized water is valuable in large scale positive recombinant bacterium screening.

Objective:To compare the Alzheimer's disease model in two species of mice by intracerebroventricular injection of β-amyloid peptide 25-35(Aβ_(25～35)).Method:The step down test and Morris water-maze were used to investigate the influence of the mice's learning and memorizing ability after intracerebroventricular injection of β-amyloid.Results:Normal Kunming mice and BALB/c mice had no significant difference in step down test, but the space cognitive ability of Kunming mice was better than that of BALB/c mice. The learning response in step-down test and Morris water-maze is no influence in male, female BALB/c mice and female Kunming mice. Marked differences were observed in male Kunming mice in acquisition, performance and reversal of a place learning response in a Morris water-maze.Conclusion:The male Kunming mice is much better than female Kunming mice and male, female BALB/c mice in preparing Alzheimer's disease animal model by intracerebroventricular injection of Aβ_(25～35), and is the most suitable animal in this experiment.

OBJECTIVETo investigate the effect of fetal bovine serum (FBS) on expression of pigment epithelium-derived factor (PEDF) in normal epidermal keratinocytes and dermal fibroblasts.

METHODSKeratinocytes and fibroblasts were incubated with 10% FBS. PEDF protein level in the cells was determined by immunofluorescence and Western blot.

RESULTSPEDF was localized mostly in the cytoplasm,while some in the nuclei. The distribution of PEDF in cytoplasm was in a granular pattern. 10% FBS increased the expression of PEDF both in keratinocytes and fibroblasts,but histamine and Phorbol 12-myristate 13-acetate (PMA) did not interfere the distribution of PEDF in cells.

CONCLUSION10% FBS can upregulate expression of PEDF in epidermal keratinocytes and dermal fibroblasts.

Animals ; Cattle ; Cells, Cultured ; Epidermis ; cytology ; metabolism ; Eye Proteins ; genetics ; metabolism ; Fetus ; Fibroblasts ; cytology ; metabolism ; Keratinocytes ; cytology ; metabolism ; Nerve Growth Factors ; genetics ; metabolism ; Serpins ; genetics ; metabolism ; Serum ; physiology ; Skin ; cytology ; metabolism ; Up-Regulation

OBJECTIVETo evaluate the oncological and functional outcomes in patients undergoing supracricoid partial laryngectomy (SCPL).

METHODSForty-three cases underwent supracricoid partial laryngectomy with functional reconstruction in stage T1b-T4. In these cases, 16 were supraglottic cancers, 21 were glottic cancers, and 2 were subglottic cancers. Three types of supracricoid partial laryngectomy (cricohyoidopexy CHP, cricohyoidoepiglottopexy CHEP and tracheocricohyoidoepiglottopexy TCHEP) were employed accordingly.

RESULTSThe 3- and 5-year accumulative survival rates were 90.7% and 83.7% respectively (Kaplan-Meier method). Median follow-up time was 57 months. Decannulation rate was 95.3% (41/43) in those. The mean time of decannulation was 14 days in patients preserved both cricoarytenoid units and 43 days in those only preserved one cricoarytenoid unit. The removal of epiglottis increased the risk of aspiration and prolonged time of decannulation (P < 0.05).

CONCLUSIONSSupracricoid partial laryngectomy for selected laryngeal cancer is feasible. The patients can gain satisfied survival rate and quality of life.

Aged ; Carcinoma, Squamous Cell ; mortality ; surgery ; Cricoid Cartilage ; surgery ; Female ; Humans ; Laryngeal Neoplasms ; mortality ; surgery ; Laryngectomy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome

Paeoniflorin (PF) is the chief active component of paeonia, with diverse pharmacological actions and wide application. Recently, the effect of PF on nervous system has attracted increasingly more attention. According to current study findings, PF can ameliorate the decline of memory and learning capacities in many dementia model animals, and have effect in protecting the cerebral ischemia injury, treating Parkinson's disease, reliving pain and improving neural synapse plasticity. Thought its mechanism has not been clarified, current findings show that adenosine A1 receptor plays an important role, while M cholinergic receptor, opiate receptor, calcium ion channel and NF-KB may also play a part in paeoniflorin's effect on nervous system.
Animals ; Benzoates ; pharmacology ; Bridged-Ring Compounds ; pharmacology ; Calcium Channels ; metabolism ; Glucosides ; pharmacology ; Learning ; drug effects ; Memory ; drug effects ; Monoterpenes ; NF-kappa B ; metabolism ; Nervous System ; drug effects ; metabolism ; Receptor, Adenosine A1 ; metabolism

OBJECTIVETo investigate the effect of adenovirus-mediated endostatin gene transfer on transplanted lung cancer in mice and its mechanism of action.

METHODSTransplant tumor model was induced by subcutaneous inoculation of 2 x 10(6) Lewis lung cancer (LLC) cells into the back of C57BL/6 mice. The mice were treated by intratumoral injection of 2 x 10(9) pfu Ad-mEndostatin. The expression of endostatin in situ and its maintaining time were detected by immunohistochemistry and Western Blot, respectively. The endostatin level in serum was determined by ELISA . The inhibition of tumor growth and changes of survival were recorded and the microvessel density (MVD) was determined by histochemical stainingwith CD31 and CD105 antibodies. The tumor apoptosis was observed by electron microscopy.

RESULTSIn comparison with controls, intratumoral injection of Ad-mEndostatin significantly inhibited the tumor growth and metastasis, and prolonged the survival rate of mice (P < 0.05). Strong positive expression of mEndostatin was seen in the tumor tissue after injection of Ad-mEndostatin, immunhistochemically ostained by mouse endostatin monoclonal antibody, while the control groups showed only very low expression or absence. Serum endostatin concentration was 1540 +/- 560 ng/ml at the second week of administration, the expression of endostatin diminished a month later. The microvessel density (MVD)) decreased from 42.4 +/- 4.8 to 10.5 +/- 3.2 per x 200 magnificetion microscopic field by CD10 staining and from 68.5 +/- 4.5 to 37.5 +/- 4.6 by CD31 staining, respectively (P < 0.05). More apoptotic tumor cells were seen under the transmission electron microscope.

CONCLUSIONEndostatin gene therapy mediated by adenoviral vector efficiently induces expression of endostatin in vivo, and inhibits the growth and metastasis of tumor. It is concluded that its action is targeted to tumor neovasculature and the mechanism is inhibition of tumor angiogenesis.

Adenoviridae ; genetics ; Angiogenesis Inhibitors ; genetics ; metabolism ; therapeutic use ; Animals ; Carcinoma, Lewis Lung ; metabolism ; pathology ; therapy ; Cell Line, Tumor ; Endostatins ; genetics ; metabolism ; therapeutic use ; Genetic Therapy ; Genetic Vectors ; Male ; Mice ; Mice, Inbred C57BL ; Microvessels ; pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; pathology ; Random Allocation ; Transfection ; Tumor Burden

OBJECTIVETo investigate the effects of phytoestrogens (daidzein and genistein) on the testosterone production of rat Leydig cells and the possible mechanisms.

METHODSPrimary Leydig cells were obtained from 3-month old male SD rats using discontinuous Percoll density gradient centrifugation. The effects of phytoestrogens at various concentrations were evaluated by ELISA, with hCG as the positive control. The mRNA expression of P450 side-chain cleavage enzyme (P450scc) was analyzed by semi-quantitative RT-PCR.

RESULTSGenistein at 0.1 micromol/L obviously promoted the secretion of testosterone and upregulated the mRNA level of P450scc. At a higher concentration of 5 micromol/L, however, both daidzein and genistein significantly inhibited the testosterone production of Leydig cells (P > 0.05).

CONCLUSIONGenistein can promote the testosterone production of Leydig cells at a low concentration (0.1 micromol/L), but both daidzein and genistein can inhibit it at a higher concentration ( >5 micromol/L).

Animals ; Cells, Cultured ; Genistein ; pharmacology ; Isoflavones ; pharmacology ; Leydig Cells ; drug effects ; secretion ; Male ; Phytoestrogens ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Testosterone ; biosynthesis

OBJECTIVETo investigate whether there is endogenous neural stem cell proliferation and whether these proliferated neural stem cells represent neural plasticity in the adult rats after cerebral infarction.

METHODSCerebral infarction models of rats were established and the dynamic expression of bromodeoxyuridine (BrdU), BrdU/polysialylated neural cell adhesion molecule (PSA-NCAM) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark dividing neural stem cells. PSA-NCAM was used to mark the plasticity of neural stem cells.

RESULTSCompared with controls, the number of BrdU-positive cells in the subventricular zone (SVZ) and hippocampus increased significantly at 1st day after cerebral infarction (P < 0.05), reached maximum at 7th day, decreased markedly at 14th day, but it was still elevated compared with that of the controls (P < 0.05). The number of BrdU-labeled with PSA-NCAM-positive cells increased significantly at 7th day (P < 0.05), reached maximum at 14th day, markedly decreased at 28th day, but it was still elevated compared with that of the controls (P < 0.05). It was equal to 60% of the number of BrdU-positive cells in the same period.

CONCLUSIONCerebral infarction may stimulate the proliferation of endogenous neural stem cells in situ and most proliferated neural stem cells represent neural plasticity.

Animals ; Bromodeoxyuridine ; metabolism ; Cell Proliferation ; Cerebral Infarction ; metabolism ; pathology ; Cerebral Ventricles ; pathology ; Hippocampus ; pathology ; Male ; Neural Cell Adhesion Molecule L1 ; metabolism ; Neuronal Plasticity ; Neurons ; metabolism ; pathology ; Rats ; Rats, Wistar ; Sialic Acids ; metabolism ; Stem Cells ; metabolism ; pathology