Background:As an important catalytic subunit of telomerase,human telomerase reverse transcriptase (hTERT)plays an important role in the development and progression of many cancers including gastric cancer.It has been reported that several single nucleotide polymorphisms (SNPs)of hTERT had varying degrees of association with risk of neoplasms. Aims:To study the correlation between SNPs of hTERT rs2853676 and rs2853677 and susceptibility to gastric cancer. Methods:Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotypes of rs2853676 and rs2853677 of hTERT in 297 gastric cancer patients,105 atrophic gastritis and 402 controls. Helicobacter pylori (Hp)infection was detected by pathological examination and 13 C-urea breath test.Results:Frequency of AA genotype of rs2853676 was significantly higher in gastric cancer group when compared with control group (15.2%vs.6.5%,P =0.01).The risk of gastric cancer in AA genotype carriers increased 2.47-fold (95% CI:1.46-4.16) when compared with GG carriers.No significant differences in the frequencies of CC,TC and TT genotypes of rs2853677 were found among gastric cancer patients,atrophic gastritis patients and controls.Hp infection rates in atrophic gastritis group and gastric cancer group were significantly increased than those in controls (64.8%,56.9% vs.40.3%,P all <0.01),OR were 2.73 (95% CI:1.74-4.26),1.96 (95% CI:1.44-2.67),respectively.Logistic regression analysis showed that there was no significant interaction between Hp infection and gene mutation.Conclusions:Polymorphism of hTERT gene rs2853676 may play a role in susceptibility to gastric cancer,and Hp infection may not be involved in the increase of risk of gastric cancer caused by hTERT gene polymorphism.

Objective:To investigate the expression of maternally expressed gene 3 (MEG3), a long non-coding RNA gene, in gastric can-cer tissues;determine the relationship of MEG3 with the prognosis of gastric cancer;and explore the relationship between MEG3 and apoptosis-associated protein P53 as well as murine double minute 2 (MDM2). Methods:Fifty-five consecutive patients with gastric cancer admitted to Qingdao Municipal Hospital for surgical treatment from September 2012 to June 2013 were included in this study. Gastric cancer and paired normal tissues were collected. The expression of MEG3 was tested through real-time quantitative poly-merase chain reaction (qRT-PCR). Western blot analysis was used to detect the expression of P53 and MDM2 in gastric cancer and eval-uate their correlations with MEG3. Results:The expression of MEG3 decreased in cancer tissues (7.98±0.19) relative to the correspond-ing normal tissues (9.47±0.18) (P<0.05). P53 and MDM2 showed negative relationships in the gastric cancer and normal tissues. A posi-tive relationship was found between P53 and MEG3 (r=0.591, P<0.05), whereas a negative relationship was found between MDM2 and MEG3 (r=?0.346, P<0.05). The median survival time was significantly prolonged in patients with high MEG3 expression compared with patients with low MEG3 expression. Conclusion:MEG3 exerts an inhibiting effect on the development of gastric cancer. MEG3, P53, and MDM2 may have important relationships in the biological mechanisms of gastric cancer development. Detecting the expression level of MEG3 may be useful for the prognosis of gastric cancer.

OBJECTIVETo explore the effect of ligustrazine on the migration of bone marrow mesenchymal stem cells (BMSCs) and protein expressions of matrix metalloproteinase-2 and-9 (MMP-2 and MMP-9) in vitro.

METHODSBMSCs were in vitro isolated and cultured using whole bone marrow adherent method, and phenotypes [surface positive antigens (CD29 and CD90) and negative antigens (CD34 and CD45)] identified using flow cytometry. BMSCs were divided into the blank control group, 25, 50, 100 µmol/L ligustrazine group, and the GM6001 group (100 µmol/L ligustrazine +MMPs inhibitor GM6001 ). The migration of BMSCs was tested by Transwell chamber test and wound healing assay after treated with ligustrazine for 24 h. The protein expressions of MMP-2 and MMP-9 were detected by Western blot.

RESULTSThe third passage BMSCs grew well in uniform morphology. The expression rate of CD29, CD90, CD34, and CD45 was 96.9%, 97.3%, 0.2%, and 3.0%, respectively. Compared with the blank control group, the number of migrated cells and relative distance of cell invasion increased, and the protein expressions of MMP-2 and MMP-9 were elevated in each ligustrazine group (P < 0.05, P < 0.01). Compared with 100 µmol/L ligustrazine group, the number of migrated cells and relative distance of cell invasion decreased in 25 and 50 µmol/L ligustrazine groups and the GM6001 group (P < 0.01). Protein expression of MMP-2 decreased in 25 and 50 µmol/L ligustrazine groups (P < 0.01).

CONCLUSIONLigustrazine could promote the migration of BMSCs in vitro, and its mechanism might be related to up-regulating expression levels of MMP-2 and MMP-9 protein.

Cell Movement ; Cells, Cultured ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Pyrazines ; pharmacology ; Up-Regulation

The aim of this study was to improve the stability and cover the unpleasant odor of valerian oil by preparation of beta-cyclodextrin inclusion complex. The preparation method was established based on the yield of inclusion complex and entrapment efficiency of valerian volatile oil. After that, the formulation and processing parameters were optimized by uniform design table. The formations of inclusion complex were validated by DSC and X-RD method. The stability of valerian oil beta-cyclodextrin inclusion was studied under stressed conditions. In conclusion, relatively high yield of inclusion complex and entrapment efficiency were obtained by saturated solution-ultrasonication method. Inclusion complex yield and entrapment efficiency of the valerian oil were (84.78 +/- 3.23)% and (86.23 +/- 2.48)%, which were prepared under the optimized conditions, respectively. The results of DSC and X-RD were indicated the formation of inclusion complex. The stability of test showed that the valerian oil-beta-cyclodextrin inclusion complex was improved significantly.
Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Odorants ; Oils, Volatile ; chemistry ; Valerian ; chemistry ; beta-Cyclodextrins ; chemistry

Melittin exhibits high antibacterial potency against drug-resistant bacteria. However, the clinical utility of melittin is limited by its serious hemolytic activity. Thus, the need for developing novel melittin analogues with high antimicrobial activity and low hemolytic activity has grown. We designed, synthesized, and evaluated 20 novel melittin analogues with varying hydrophobic, polar or positively charged amino acids. The results showed that 8 compounds had antimicrobial activity (MIC: 1-4 μg·mL^-1) against gram-positive pathogens equal to or better than that of melittin, and 16 compounds had low hemolytic activity (HC₅₀ ≥ 11.9 μg·mL^-1). Compounds 13 (MIC: 2-4 μg·mL^-1) and 15 (MIC: 1-2 μg·mL^-1) showed equal or better antimicrobial activity against both susceptible and resistant strains of Staphylococcus aureus and Enterococcus faecium compared to melittin (MIC: 4 μg·mL^-1). Compound 13 (HC₅₀: 24.0 ± 4.3 μg·mL^-1) displayed noticeably decreased hemolytic activity compared to melittin (HC₅₀: 5.3 ± 0.4 μg·mL^-1). This work established a base for further study on the structure-activity relationships and structure-toxicity relationships of melittin.

The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF.
Animals ; Cell Line ; Chickens ; Chorioallantoic Membrane ; blood supply ; Disease Models, Animal ; Genetic Therapy ; Humans ; Male ; Myocardial Infarction ; metabolism ; pathology ; therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.
ADP Ribose Transferases ; chemistry ; metabolism ; pharmacology ; Amino Acid Sequence ; Blotting, Western ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Exotoxins ; chemistry ; metabolism ; pharmacology ; Humans ; Interleukin-6 ; chemistry ; metabolism ; pharmacology ; Molecular Sequence Data ; Pseudomonas aeruginosa ; genetics ; metabolism ; Recombinant Fusion Proteins ; chemistry ; metabolism ; pharmacology ; Sequence Analysis, Protein ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo prove analgesia and sedative effect of adjuvant anesthesia with magnetic auricular point-sticking on abdominal gynecological operation and its effect on postoperative recovery of body function.

METHODSNinety-two patients with abdominal gynecological operation were randomly divided into 3 groups. The auricular point-sticking group (APS group, n=31) was pasted and pressed by plasters with magnetic beads at bilateral Shenmen, Pizhixia (subcortex), Zigong (uterus) and Penqiang (pelvic cavity), etc. the night before operation. The placebo group (n=31) was pasted by plasters without magnetic beads. The blank group (n=30) was given no intervention. The mental and gastrointestinal functional changes before and 3 days after the operation were observed.

RESULTSAs compared with those in the control group and the blank group, the postoperative score of Self rating Anxiety Scale (SAS) was less (25.5 +/- 0.81 vs. 28.9 +/- 3.19, 28.3 +/- 2.36, both P < 0.01), with lower-dose of Innovar [(2.5 + 1.1) mL vs. (3.4 + 1.8) mL, (3.2 + 1.6) mL, both P < 0.05], earlier exsufflation after the operation [(34.2 + 12.1) h vs. (46.3 + 10.9) h, (43.2 + 14.8) h, both P < 0.01] and higher level serum of beta-endorphin before and after the operation in the APS group (all P < 0.05).

CONCLUSIONThe magnetic auricular point-sticking has sedative, analgesic and function-regulating effects on the abdominal gynecological operation.

Acupuncture Analgesia ; Acupuncture, Ear ; Adjuvants, Anesthesia ; administration & dosage ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; drug therapy ; physiopathology ; therapy ; Postoperative Period ; Recovery of Function ; Young Adult

OBJECTIVETo discuss the mechanism of traditional Chinese medicines reducing phlegm and resolving masses in treatment of iodine deficiency-induced goiter by observing the expression of growth factors and the balance-regulating mechanism of proliferation and apoptosis.

METHOD180 four-week-old Wistar rats were selected to establish the iodine deficiency model. After the modeling, the rats were randomly divided into six groups: the normal control group, the model control group, the iodine group, the phlegm compound group, the L-T4 group and the phlegm compound and L-T4 group. At the 21st day and 77th day after administration, 15 rats in each group were killed to collect specimens. Doses were calculated and adjusted according to body surface area and body weight. TT3, TT4 radioimmunoassay, TSH, immunoradiometric method were adopted. Fas, FasL and PCNA protein expressions are detected using immunohistochemical methods.

RESULTCompared with the normal group and the model group, the expressions of fas and FasL in the phlegm Group significantly increased, the expressions of fas and FasL in the phlegm and L-T4 group were also increased significantly. The expression of fas in the L-T4 Group was significantly lower than that of the L-T4 group and the phlegm compound and L-T4 group. Compared with the normal group, the expression of PCNA of the phlegm group and the phlegm and L-T4 group was significantly lower. Compared with the model group, the expression of PCNA of the iodine group, the phlegm groups and the phlegm and L-T4 group were significantly lower. Compared with the normal group, the expression of VEGF in the iodine group significantly decreased after treatment. Compared with the iodine group, the expression of VEGF in the phlegm group and the L-T4 group significantly reduced. Compared with the normal group, the expression of TGF-beta1 in the model group and the phlegm group significantly increased. Compared with model group, the expression of TGF-beta1 in the iodine group significantly reduced. Compared with the phlegm group, the expression of TGF-beta1 in the phlegm compound and L-T4 group was significantly reduced.

CONCLUSIONTraditional Chinese medicines reducing phlegm and resolving masses can completely recover goiter by promoting apoptosis of thyroid cells, inhibiting their proliferation and the expression of growth factors and enhancing the expression of TGF-beta, without causing injury on thyroid cells.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Goiter ; drug therapy ; genetics ; metabolism ; Humans ; Male ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Rats ; Rats, Wistar ; Thyroid Hormones ; secretion ; Vascular Endothelial Growth Factor A ; genetics ; metabolism