Objective:To predict the B cell epitope for MUC1 antigene and screen the mimic epitope of MUC1 from random phage dispay peptide library. Methods:In order to predict the B cell epitope for MUC1 antigen,the secondary structure and antigenicity was analysed with various methods. The purified Ma695 Ab was used to screen in phage random 12 peptide library. The positive clones were identified by sandwich ELISA and competitive inhibiton assay. Results:17 distinct antigenic epitope regions in MUC1 were identified by computation. 14 positive clones were acquired after 3 rounds of screening. Amino acid sequences deduced from DNA sequences showed four different sequences:KHYDPFHHRMPQ,QADTARSVALAG,VPSKPDLHVRSI and MTPIHYWNHNRV. The inhibitory assay showed that the 4 mimic epitope peptides displaying on the phage surface could effectively inhibit the combination of antibody with antigen and the inhibitory rates of each mimic epitope were 50% higher than that in the controls. Conclusion: Prediction of the B cell epitope for MUC1 can provide a basic clues for studies on structure and function of MUC1. The results indicated that KHYDPFHHRMPQ,QADTARSVALAG,VPSKPDLHVRSI and MTPIHYWNHNRV are the mimotopes which could mimic the epitope of MUC1.

Objective To study the changes of tartrate resistant acid phosphatase in the serum of patients with bone metastasis in pros-tate cancer,and to explore its clinical significance for bone metastasis diagnosis and prediction of prostate cancer. Methods Seventy-eight cases of prostate cancer and 40 cases of benign prostatic hyperplasia patients as the research object,at the same time,40 healthy young men as controls,were divided into the group of patients with prostate cancer bone metastases (group A,n=41),prostate cancer patients with no bone metastasis group (group B,n=37),benign prostatic hyperplasia patient group (group C,n=40),healthy control group (group D,n=40). Determination of serum TrACP levels in patients with the immunoassay using double antibody sandwich ELISA,combined with pathologi-cal grade,Gleason score,PSA,ALP and ALT were statistically analyzed. Results The serum TrACP concentration in patients with bone me-tastasis significantly increased,with significant difference compared with the other groups. The concentration of serum TrACP and PSA levels showed a positive correlation. The area under the ROC curve ( AUC) was higher than that in ALT,and the ROC curve of cross TrACP and PSA,suggesting that high value in the diagnosis of bone metastasis. Conclusion The detection of TrACP has directly diagnosis and predic-tive value for prostate cancer with bone metastasis,the serum TrACP content monitoring in patients with prostate cancer has valuable clinical significance in understanding the growth progression status of prostate cancer,judgment and prediction of bone metastases.

BACKGROUND:At present, occlusion of the drainage tube was commonly used to reduce the drainage volume after total hip arthroplasty so as to promote the incision healing and hip function rehabilitation. However, the occlusion time is a problem deserving further investigations. OBJECTIVE:To research the effects of temporarily clamping drainage tube on drainage volume in early stage after total hip arthroplasty. METHODS:From January to October 2013, 112 patients received unilateral total hip arthroplasty in the First Ward, Department of Orthopedics, First Affiliated Hospital, Zhengzhou University in China. They were randomly divided into four groups according to the admission time (n=28):occlusion of the drainage tube for 2, 4 and 6 hours and without occlusion of the drainage tube. Al tubes were pul ed out in postoperative 48 hours. Moreover, 48-hour postoperative hemoglobin and drainage volume, the healing of incision and the score of hip joint function when patients were fol owed up at 1.5 months postoperatively were recorded accurately. RESULTS AND CONCLUSION:48-hour hemoglobin drop level and 48-hour incision drainage volume were highest in the non-occlusion group, fol owed by 2-hour occlusion group, 4-hour occlusion group and 6-hour occlusion group (P<0.05). However, no significant differences in the healing time of postoperative incision and the Harris score of hip joint function when patients were fol owed up at 1.5 months postoperatively were detected among groups (P>0.05). Incision infection and tension split were not seen in each group. Six cases affected subcutaneous ecchymosis and swel ing accompanied by pain and four cases suffered from the venous plexus thrombosis of the calf muscle in the 6-hour occlusion group. One case experienced fat liquefaction separately in the 4-hour occlusion group and non-occlusion group. These results suggested that temporary occlusion of the drainage tube in the early stage of total hip arthroplasty could reduce the drainage volume of incision. The suitable time of clamping drainage tube was 4 hours, and there were no adverse effects on healing of incision and recovery of hip function.

Purpose:To study the reversal effects on multidrug resistance in multidrug-resistent bladder cell lines BIU-87/ADM. Methods:The degree of drug-resistence was detected by MTTmethod; the concentration of ADM in cells BIU-87 and BIU-87/ADM was detected after the cells were treated by carvedilol and ADM. Results:Pretreatment by carvedilol increased the concentration of ADM in BIU-87/ADM cells, ,and the concentration was much higher than that in those cells treated by ADM only. Conclusions:Carvedilol can enhance the cytotoxic effect of ADM,and can reverse the multidrug resistance in bladder cancer lines BIU-87/ADM.

Virtual reality simulators and surgical demonstration were used in laparoscopic teaching combining teacher' guiding with students' practice to improve the teaching effectiveness hecause of the limitations of the traditional apprenticeship teaching methods.The new method was applied for 2 cycles and for 6 weeks.The results showed that the teaching methods of virtual reality simulators combined with surgical demonstration can significantly improve the effectiveness of laparoscopic teaching and shorten the initial learning curve,therefore it is worth promoting in laparoscopic operation training and teaching.

Objective To explore the influences of decorin ( DCN) gene down-regulating on biological behavior in human lung adenocarcinoma A 549 cell line.Methods Chemically synthesis in vitro targeting DCN -siRNA was transfected into human lung adenocarcinoma A 549 cells by LipofectamineTM 2000 .The gene expres-sion of DCN was detected using Real-Time PCR ;The protein expression of DCN was investigated using Western blot;Checkout DCN-siRNA effects on lung adenocarcinoma cancer cell proliferation was detected by CCK -8;The migration and invasion ability were determined by Transwell assay .Results DCN-siRNA was successfully transfected into human lung adenocarcinoma A 549 cells.Real-Time PCR results showed that DCN mRNA ex-pression level significantly decreased ,compared with untransfected group ,negative control group .Western blot re-sults showed that DCN-siRNA transfection inhibited DCN protein expression level;CCK-8 results showed that the proliferation was enhanced in the DCN -siRNA group as compared with untransfected group ,as well as the negative control group and empty vector group .Transwell assay results showed the invasion of A 549 cells in DCN-siRNA group was significantly enhanced as compared with untransfected group [(22.6 ±1.14) vs.(5.2 ± 0.84)].Wound-Healing assay results revealed that the cell repairing rate was markedly increased in DCN -siRNA group.A549 cells were close to complete repair after 48 hours.Apoptosis was not significant in DCN -siR-NA group as compared with untransfected group (P=0.214).Conclusion Down-regulation the expression of DCN gene by DCN-siRNA may enhance the ability of invasion ,migration and proliferation of A 549 cells without affecting apoptosis ,which provides a new evidence of function for advancing research of lung cancer .

Purpose:To study the expression of TopoⅡ gene in multidrug resistant cells of human bladder cancer. Methods:The degree of drug resistance was detected by MTT method ;the expression of TopoⅡ gene in cell lines BIU 87/ADM and BIU 87 was detected with reverse transcriptase polymerase chain reaction. Results:The cell lines BIU 87/ADM were 56.4 times more resistant to ADM than the cell lines BIU 87;the expression of TopoⅡ gene was poorly positive in BIU 87/ADM but strongly positive in BIU 87 cells. Conclusions: The decreased expression of TopoⅡ gene in BIU 87/ADM cells might contribute to the development of multidrug resistance of human bladder cancer.

Purpose:To study antigenic epitopes and expression of P glycoprotein.Methods:The secondary structure and surface properties of P glycoprotein was studied by many methods , designed a pair of primers to amplify DNA with PCR. The product was inserted into a pGEM T vector, sequenced and cloned into pGEX 2T vector, established a high expression recombinant vector named pGEX Pgp,which was transferred into DH5? and expressed , identified and purified. Results:Antigenic epitopes in extracellular fragment of P glycoprotein were identified in amino acid residues 82 115(I),741 760(IV) and 800 816(V). SDS PAGE analysis indicated that the expressed protein was about 30?10 3 (30 kD).Conclusions:High level expression of the target gene fragment was established for preparation of its antibody and biospanning its specific peptide using random phage library , which became the basis for the targeting treatment of MDR. [

Objective To study the expression of the extracellular fragment of P glycoprotein in the bacteria. Methods According to the P glycoprotein antigen analyses and mdr1 gene structure, a couple of primers were designed to amplify the extracellular DNA fragment of the protein with PCR. The product was inserted into pGEM T vector followed by sequencing. The sequencing verified vectors were cloned into pGEX 2T to get a highly expression recombinant vector pGEX Pgp. The recombinant was transferred into E. coli DH5?, and its expression, identification and purification were studied. Results The highest level of recombinant expression was achieved at 4 h after IPTG induction. SDS PAGE analysis indicated that the expressed protein was about 30?10 3. Conclusion The target gene fragment expression at high level is established for the preparation of its antibody and biospanning of its specific peptide by using random phage library, founding a basis for the targeting treatment of multidrug resistance.

With the modernization of traditional Chinese medicine (TCM), classic extraction methods such as sol-vent extraction, steam distillation, squeezing method, sublimation method have become much more difficult in sat-isfying needs of the development of the society as a result of low extraction rate of effective components, low per-centage impurity clearance. Therefore, in recent years the new extraction techniques with the advantages of green environmental protection , conditional stability , high extraction rate , strong biological activity , have been widely used in TCM extraction. These new extraction techniques have already shown each of its strength. The application progress on new extraction technology of Chinese medicine polysaccharide, both in standalone application and in collaboration application, were discussed in this article, so as to make new technologies be applied more reason-ably and effectively in Chinese medicine polysaccharide components.