Objective To detect the mutation of A379,we develop a TaqMan fluorogenic probe based amplification refractory mutation system (TaqMan-ARMS) and investigate whether the A379V variant and activity of Lp-PLA2 are the risk factors for CAD.Methods According to the amplification refractory mutation system(ARMS-PCR)combined with a TaqMan fluorogenic probe,we established and evaluated the assay teehnique of TaqMan-ARMS for measuring the genotype of variant.At the same time,we tested the aetivity of Lp-PLA2 in 395 patients with coronary heartdisease and 396 controls,whose clinical information [ages,CHO,GLU,TG,HDL,LDL,Hs-CRP,Lp (a)] were collected.Date was analyzed by using Independent-samples t test,Chi-square test,One-Way ANOVA,Binary Logistic Regression.Results CAD group had significantly higher Lp-PLA2 activity than the controls(31.51 nmol · ml-1 · min-1 ＞21.31 nmol ·ml-1 · min-1,F =16.40,P ＜ 0.001).Comparing the highest quartile of Lp-PLA2 activitv to the bottom quartile,OR was 7.50 (95％ CI:2.34-24.05) after adjustment for various traditional cardiovascular risk factors,including ages,sex,CHO,TG,Hs-CRP,Lp (a) and GLU; the genotype VV of A379V was associated with higher risk of CAD (OR =2.95; 95％ CI:1.22-7.15,P ＜ 0.05).Conclusions The TaqMan-ARMS real time PCR technique is established to analyze A379V genotype.Lp-PLA2 activity is significantly higher in CAD group and is a risk factor for CAD; the genotype VV of A379V is also a risk factor for CAD.

Objective To study the effect of listening to music with headphone or background music on pain reduction of patients undergoing colonoscopy. Methods 180 patients undergoing colonoscopy were randondy selected as the headphone group,background music group and the control group.The headphone group listened to relaxed music with headphone after measurement of vital signs.The background music group listened to relaxed music played by CD after measurement of vital signs.The control group did not listen to music.Pain degree was measured by pain intensity number scale during colonoscopy.Heart rate,blood pressure,saturation of blood oxygen and appraisal to the music by the former two groups were observed.Results Pain degree of the headphone group and the background music group was lower than that of the control group(P＜0.05).Pain degree of the headphone group was lower than that of the background music group with no significant difference(P＞0.05).Heart rate of the control group after colonoscopy was faster than before colonoscopy(P＜0.05).Conclusions Relaxed music can alleviate pain of patients undergoing colonoscopy.Listening to background music is a convenient and economical method for alleviation of pain without any side effects for patients undergoing colonoscopy.

Objective To establish an AGS cell line that stably expressing miR?126 and to study the effect of miR?126 on the proliferation and metastatic abilities of the AGS cell line in vitro. Methods AGS cells were infected by lentivirus with Lv?has?mir?126. After confirmation by RT?PCR ,CCK?8 and clone formation assays were used to evaluate the effect of miR?126 on AGS cell growth. Transwell migration and invasion assays were used to evaluate the effect of miR?126 on metastasis of AGS cells. Results We verified correct construction of recombinant AGS cells. RT?PCR confirmed mRNA levels of miR?126 existed significantly differences among the recombinant cell lines (P< 0.05). Proliferation assays and clone formation assays did not show a remarkable growth suppression in AGS?mir?126 cell line. However,transwell assay showed a notable acceleration in AGS?mir?126(P< 0.05). Conclusions We successfully constructed recombinant AGS cell line with stably high miR?126 expression level. MiR?126 could facilitate the metastasis of AGS cell in vitro.

Objective:This study aimed to identify the morphology of mast cells by using a modified toluidine blue staining scheme,so as to provide a powerful reference for the experimental basis research of mast cells.Methods:Bone marrow-derived mast cells were induced in vitro.After 4 weeks,the cells were collected,fixed,and stained.Mast cells were fixed at different temperature during different time.The optimum condition was determined by comparing the effects of toluidine blue staining.Results:Bone marrow cells were induced to differentiate into mast cells by SCF and IL-3 in vitro.When mast cells were stained with modified toluidine blue staining,the staining effect was better.Mast cells were round or oval and the cell membrane was complete and the cytoplasm was filled with a large number of purple particles.Conclusion:In this study,we successfully applied a modified toluidine blue staining method to mast cells cultured in vitro.The results showed that the condition at 37 ℃ full fixation with staining could reduce the degeneration of mast cells.This method was easy to operate with good stability.It was suitable for the morphological observation of mast cells cultured in vitro.

Objective To examine how ischemic time under common temperature affects acute rejection by using a rat vascularized skin transplantation model.Methods Vascularized groin flaps were transplanted from BN to Lewis rats with 1,2,3 and 4 h of ischemic time (Isc-1 h,2 h,3 h,4 h groups) under common temperature,and the allografts in each group were evaluated daily.Groin flaps were transplanted from Lewis to Lewis rats as control group.Biopsy samples taken from the each group on the postoperative day 2-8 were graded for signs of acute rejection.Biopsy samples taken from each group on the postoperative day 6 were stained for chemokine receptor CXCR3.Results When the ischemia time was 1,2,3 and 4 h,the survival time of the grafts was (9.0 ± 1.2),(8.6 ±1.1),(8.8 ± 1.3),and (7.0 ± 0.8) days respectively.The survival time in Isc-4 h group was significantly shorter than in other groups (P＜0.05).Histological evaluation showed acceleration of activated lymphocyte infiitration in the Isc-4 h group as compared with other g.roups.Furthermore,the proportion of CXCR3 positive cells in the Isc-4 h group was (50.1 ± 8.4) ％,significantly higher than that in the other groups on the day 6 after transplantation.Conclusion When ischemic time was over 3 h,the immune response is accelerated.The acceleration is associated with the higher expression of CXCR3 in the infiltrated cells.

Background and purpose:Studies have shown that ribosomal protein L8 (RPL8) is shared by melanomas, gliomas and ovarian carcinomas. A peptide of RPL8 signiifcantly stimulated proliferation and cytokine expression of the hepler T cell clone and lymphocytes in melanoma patients. RPL8 may stimulate anti-tumor immunity, making RPL8 an attractive candidate for therapeutic intervention. In this study, we prepared DC pulsed by RPL8 (RPL8-DC) and investigate the anti-tumor effect of RPL8-DC on melanoma in mice.Methods: The recombinant protein was achieved through IPTG induction in E. coli and identiifed with Western blot. Bone marrow-derived DC was loaded with RPL8 protein. RPL8 and CD11c, CD80, MHC-Ⅰ, MHC-Ⅱmolecules on dentritic cells were monitored by lfuorescence microscope and FACS analysis, respectively. The anti-tumor effect of T cells in vitro was detected by MTT assay. Subcutaneous tumors were induced in C57BL/6 mice using B16 cells. The tumor volumes were measured after injection with RPL8-DC. Results:The puriifed protein was combined with speciifc antibodies. DCs pulsed by RPL8 were visualized under lfuorescent microscopy. CD11c, CD80, MHC-Ⅰ, MHC-Ⅱmolecules on DCs were up-regulated after stimulation with RPL8 and LPS. B16 cells were inhibited by T cells stimulated with RPL8-DC. The inhibition rate of tumor cells was 70%in RPL8-DC group when effector-to-target ratio was 30∶1, which was higher than PBS and DC groups. Inhibition of growth could be observed more signiifcantly in mice after the treatment with RPL8-DC. The mice receiving the therapy of RPL8-DC were able to survive much longer than the mice receiving control therapy. Conclusion:The DC pulsed by RPL8 protein can inhibit the growth of melanoma.

To study the chemical constituents of Veratrum dahuricum (Turcz.) Loes. f., a new aurone glycoside named as (Z)-7, 4'-dimethoxy-6-hydroxyl-aurone-4-O-β-glucopyranoside was isolated from the 95% ethanol extracts of the rhizomes and roots of Veratrum dahuricum (Turcz.) Loes. f. by repeated column chromatography on silica gel and recrystallization. Its structure was established by extensive spectroscopic analyses, and its cytotoxicities against HepG-2, MCF7 and A549 cell lines were measured in vitro.
Benzofurans ; isolation & purification ; Cell Line, Tumor ; Glycosides ; isolation & purification ; Humans ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Veratrum ; chemistry

AIM: To explore the change of the expression of cell adhesion molecule CD_ 54 and CD_ 44 in erythroleukemic K562 apoptosis cells induced by momordin from momordica charantia seeds and study the effects of cell adhesion molecule CD_ 54 and CD_ 44 on cell apoptosis induced by momordin. METHODS: After the treatment of K562 cells with appropriated concentration momordin, CCK-8 test was employed to determine K562 cells growth; flow cytometry FACScan (FITC-Annexin V staining) and electron microscopy were used to detect apoptosis; The expression of CD_ 54 and CD_ 44 were examined by flow cytometry FACScan (FITC-CD_ 54 and PE-CD_ 44 staining). RESULTS: CCK-8 test showed K562 cells growth was significant inhibited by momordin; the apoptosis was detected by cell morphology and flow cytometry FACScan (FITC-Annexin V) in K562 cells after treatment by appropriated concentration momordin. The expressions of CD_ 54 and CD_ 44 in momordin treated K562 cells were 18.62 % and 1.32 % respectively, and in negative momordin treated K562 cells were 0.25 % and 0.17 % respectively, and momordin could up-expresses the protein of CD_ 54 18.37 % and CD_ 44 1.15 %. CONCLUSION: Momordin can markedly induce the K562 cell to apoptosis. The up-expressions of CD_ 54 exist in the process of apoptosis induced by momordin. The change of cell adhesion molecule maybe one of the key factors in the mechanisms of apoptosis induced by momordin, and its mechanism maybe involve in adhesion-dependent apoptosis.

Objective To investigate the allergization of milk high molecular weight proteins. Methods Thirty cas?es of patients with serum allergic to milk were selected. Their skin prick tests were positive. Results of serum specific IgE (sIgE) test were positive and≥1+. One case of healthy control with negative sIgE test and without history of allertgy, was in?cluded in this study. The serum samples were collected and frozen at-20℃. Sephadex G200 gel chromatography was used to obtain milk high molecular weight proteins. Western blot and ELISA methods were used to detect milk high molecular weight proteins and the activity of serum sIgE. Results Results of SDS-PAGE showed that high molecular weights proteins displayed by Sephadex G200 gel chromatography, mainly including three bands, the molecular weight of 67 ku, 80 ku and 160 ku. Western blot analysis showed that three kinds of high molecular weights proteins can react with milk allergy serum, and the most obvious appeared near the molecular weight 67 ku band. ELISA analysis showed that the positive response rate of high molecular weight proteins with milk allergic patient’s serum was slightly higher than that ofβ-lactoglobulin (46.7%and 43.3%, respectively). Conclusion The milk high molecular weight protein components can induce specific IgE antibod?ies, which have important sensitization in the process of milk allergy.

Objective To investigate the effect of preoperative nursing intervention in the prevention of postoperative retention of urine of patients after total knee arthroplasty. Methods 128patients underwent total knee arthroplasty were divided into the intervention group and the control group (64 cases in each group) by random number table.The patients in the control group received routine nursing care and the patients in the intervention group accepted preoperative nursing intervention 3 to 5 days before surgery. The incidence of post-operation urinary retention, catheterization, satisfaction degree and compliance of the patients were compared between the two groups. Results The incidence rate of post-operation urinary retention and catheterization of the patients was respectively 15.6%(10/64) and 7.8%(5/64) in the intervention group,lower than 34.4%(22/64) and 23.4%(15/64) in the control group. The difference was statistically significant (χ2=6.00, 5.93, respectively, P<0.05). The satisfaction degree and compliance of patients were respectively 95.3%(61/64) and 75.0%(48/64) in the intervention group, higher than 85.2%(52/64) and 31.3%(20/64) in the control group,the difference was statistically significant (χ2=6.12, 24.60,respectively, P < 0.05). Conclusions Preoperative nursing intervention have good effect in the prevention of postoperative retention of urine and improving satisfaction degree of patients underwent total knee arthroplasty.