Objective: To research the expression of PTEN and its influence on biological ability in breast cancer cell in vivo. Methods: PTEN-shRNA plasmid was transtected into M231 breast cancer cells to knock down the expression of PTEN. The changes of PTEN expression, proliferation, adhesion and metastasis of PTEN knocked down cell were tested by western-blot, colony formation, adhesion and invasion assay. Results: PTEN-shRNA was successfully transfected into M231 cells and it inhibited PTEN expression efficiently.The capabilities of colony formation, migration and invasion of transfected cell were much greater than those of the controlling cell line. But the transfected cells were more difficulty in adhesion than the scrambled ones. Conclusion: PTEN genecan enhance the adhesion, but restrict the proliferation, migration of breast cancer cells in some degree, so that inhibit the development of the breast cancer. PTEN loss can be a prognostic factors for the patients with breast cancer.

BACKGROUND: Posterior internal fixation and fusion system is a main method for chronic back pain caused by intervertebral disc degeneration, but more postoperative adverse reactions occur. Dynamic stabilization system can reduce adjacent-segment degeneration, and theoretically, repair intervertebral disc degeneration.OBJECTIVE: To investigate the efficacy and safety of K-rod dynamic stabilization system in the repair of lumbar degenerative diseases.METHODS/DESIGN: We conducted a prospective, single-center, self-controlled, clinical trial at the Orthopedic Hospital of Shenyang, China. Sixty-seven patients with lumbar degenerative diseases were enrolled, and treated with K-rod dynamic stabilization system. All patients were followed for 2 years. The primary outcome was the changes in the Oswestry dysfunction index scores at baseline, 3, 6, 12 and 24 months postoperatively. The secondary outcomes were the ratio of height vertebral space to body and lumbar lordotic angle at baseline, 3, 12 and 24 hours postoperatively; the visual analogue scale scores for back pain and morphological changes in the lumbar vertebrae on x-ray preoperatively and 3, 6, 12 and 24 months after surgery; the incidence of adverse reactions at 3, 6, 12 and 24 months postoperatively.This trial has been registered at ClinicalTrials.gov (identifier: NCT03214042). The study protocol has been approved by the Ethics Committee of Orthopedic Hospital of Shenyang. All protocols will be performed in accordance with the Ethical Principles for Medical Research Involving Human Subjects in the Declaration of Helsinki. Written informed consent was provided by each patient after they indicated that they fully understood the treatment plan.DISCUSSION: This trial was designed to investigate the efficacy and safety of K-rod dynamic stabilization system for lumbar degenerative diseases, thus providing reference for its clinical application. Partial results demonstrated that the Oswestry Dysfunction Index and Visual Analogue Scale scores at 24 months postoperatively were significantly improved (P < 0.01), but the ratio of height vertebral space to body and lumbar lordotic angle did not differ significantly at different time points (P > 0.01). These results suggest that K-rod dynamic stabilization system can alleviate pain and improve lumbar function in the patients with lumbar degenerative diseases.

Objective:To evaluate the effect of iliac bone graft and radial forearm flap in the reconstrucion of maxilla.Methods:Maxilla defects were reconstructed using iliac bone graft and radial forearm flap in 4 patients.The effects were evaluated clinicaly.Results:In all the 4 cases,palatal defects resulted from maxillectomy were optimally reconstructed with non-vascularized iliac graft and radial forearm flap.The masticatory function of the upper jaw,intelligible speech,swallow and natural facial appearance were recovered.As a result,quality of life of the patients was improved.Conclusion:Iliac bone graft and radial forearm are feasible in the reconstruction of maxilla defects.

Objective To investigate the influence of impulsivity on the functions of the conflict monitoring and the conflict resolution process for heroin addicts.Methods 80 heroin addicts were drawn randomly using simple random sampling method and finished Opioid Addiction Severity Inventory ( OASI ) and Barratt Impulsivity Scale ( BIS-11) .High impulsivity group( 7 females of 22) and low impulsivity group (8 females of 21) were divided according to 27%of the total BIS-11score followed the grouping method in psychometrics.Then the event-related potentials (ERPs) technique with the classical color-word Stroop task was used to reveal the influence of impulsivity on the executive function for heroin addicts.Results ( 1) The scores of motor impulsivity(23.00±1.73) and no-plan impulsivity(27.77±3.22) were higher in high im-pulsivity group than those in low impulsivity group(19.31±2.80,23.38±3.59)(P<0.01);and there was posi-tive correlation between impulsivity and addiction severity( r=0.415, P<0.05).(2) The behavioral data showed significant Stroop interference effects in both groups(P<0.01).(3) ERPs data showed that there were significant incongruent-N450 and SP effects in the low impulsivity group,whereas incongruent N450 and SP effects disappeared in high impulsivity group ( Low impulsivity group incongruent vs congruent condition N450:(2.82±3.09)μV vs (4.51±2.77)μV, P<0.05; SP:(3.54±1.25)μV vs (2.84±1.03)μV, P<0.05;High impulsivity group incongruent vs congruent condition N450:(4.98±4.10)μV vs (3.39±3.31)μV, P<0.05;SP:(3.43±3.84)μV vs (4.66±4.53)μV, P<0.05).Conclusion The brain time-interval change of executive function such as the conflict monitoring and the conflict resolution process is influenced by the im-pulsivity levels of heroin addicts.

0.05) . The sensory block reached T7 in levobupivacaine group and T6 in bupivacaine group respectively. The motor blocked was somewhat more intense in bupivacaine group. Conclusion The efficacy and safety of epidural anesthesia with levobupivacaine and bupivacaine are comparable.

Objective:To explore the effect of siRNA targeting myeloid cell leukemia-1(Mcl-1)on the biological behavior of salivary adenoid cystic carcinoma cells.Methods:The chemically synthesized Mcl-1-siRNA was transfected into salivary adenoid cystic carci-noma SACC-2 cells.The expression levels of Mcl-1-mRNA and Mcl-1protein were examined by Real-time PCR and western blotting respectively.MTT assay,transwell chamber and flow cytometry were used to determine the effect of Mcl-1-siRNA on SACC-2 cell pro-liferation,migration and apoptosis.Results:Compared with the control group,liposome group and NC-siRNA group,SACC-2 cell proliferation rate of Mcl-1-siRNA group was obviously slowed down.48 h after transfection,the migration of SACC-2 cells in Mcl-1-siRNA group(39 ±9.0)were lower than that in control group(69 ±6.0).The apoptosis rate of Mcl-1-siRNA group(8.6%)was sig-nificantly higher than that in control group(1.9%).Conclusion:Silence Mcl-1 can inhibit cell proliferation and migration and pro-mote apoptosis of salivary adenoid cystic carcinoma cells.

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

Objective:To explore the effects of cysteine-rich 6 1 (Cyr6 1 )on biological behavior of human adenoid cystic carcinoma ACC-LM and ACC2 cells.Methods:The chemically synthesized Cyr6 1-siRNA was transfected into ACC-LM and ACC2 cells.Cell proliferation was measured by the MTT method,the invasive ability was evaluated by Transwell chamber assay,and cell apoptosis was analyzed using flow cytometry by double staining with Annexin V and propidium iodide.Results:Cyr61-siRNA significantly down-regu-lated Cyr61 protein expression in ACC-LMand ACC2 cells.Cyr61-siRNA markedly inhibited the proliferation and invasion of the cells, however,there was no significant difference in cell apoptosis between Cyr6 1-siRNA and control groups.Conclusion:Cyr6 1 promote the proliferation and invasion of adenoid cystic cancer cells.

Objective To observe the change of neuropeptide of the infant patients with post infection cough (PIC) and explore the possible pathogenesis and diagnostic value.Methods Fifty-two cases with PIC in our hospital were selected as PIC group,while thirty infant patients with bronchopneumonia as pneumonia group.Phlegm cells classification and substance P(S P),neurokinin(NKA),neurokinin (NKB),calcitonin gene-related peptide (CGRP) concentration of the patients in both groups were compared.Results Phlegm cells classification of the two groups showed that,compared with the PIC group,the neutrophils,macrophages,and lymphocytes of the infant patients in the pneumonia group were obviously higher (P ＜ 0.05).After anti-infection treatment,the neutrophils,macrophages and lymphocytes of the infant patients in the pneumonia group were obviously lower(P ＜ 0.01).The phlegm cells classification of the PIC group had not changed significantly before and after treatment (P ＞ 0.05).After treatment,the phlegm cells classification of both groups had no significant difference (P ＞ 0.05).During the progress of treatment,there was no evident change for the eosinophils (P ＞ 0.05).Neuropeptide of the two groups showed that,before treatment,the SP and CGRP of the patients in the PIC group were (538.4 ±432.2) ng/L,(123.6 ±70.2) ng/L,and that in the pneumonia group were (613.2 ± 345) ng/L,(156.2 ± 82.6) ng/L.There was no significant difference (P ＞ 0.05).After treatment,the SP and CGRP of the PIC group were (552.8 ± 421.7) ng/L,(133.5 ± 81.3) ng/L,and there was no significant difference (P ＞0.05).While the SP and CGRP of the patients in the pneumonia group were (156.2 ± 131.2) ng/L,(741.2 ± 35.4) ng/L,they were obviously lower (P ＜ 0.01).There was statistical difference compared with the PIC group after treatment (P ＜ 0.01).The NKA and NKB had not changed significantly before and after treatment.Conclusion Airway neurogenic inflammation mediated SP and CGRP plays a key role in the acute and chronic infections.The increase of SP and CGRP is closely related to the morbidity of PIC.So SP and CGRP is a reference index for early detection of PIC.

AIM:To explore the effects of miR-21 on biological behavior of colon cancer cells and their sensi-tivity to epidermal growth factor receptor monoclonal antibody cetuximab .METHODS:Lentiviral vectors were constructed to generate up-and down-regulations of miR-21 lentiviruses (LV-miR-21 and LV-anti-miR-21, respectively), and the cor-responding negative control viruses (LV-miR-21 NC and LV-anti-miR-21 NC, respectively) were also constructed.The vi-ruses were used to infect human colon cancer RKO cells .The changes of the miR-21 expression level , the cell prolifera-tion, the colony-forming ability, the cell apoptosis and the sensitivity of the cells to cetuximab were detected by real -time PCR, MTT assay, soft agar colony assay , flow cytometry and CCK-8 assay.RESULTS: The lentivirus titers of LV-miR-21, LV-miR-2 NC, LV-anti-miR-21 and LV-anti-miR-21 NC were 3.0 ×1012 TU/L, 6.0 ×1011 TU/L, 2.0 ×1012 TU/L and 8.0 ×1011 TU/L, respectively.The infection efficiency was over 80% by the observation of green fluorescence .The miR-21 expression level , the cell proliferation , and the colony-forming ability in LV-miR-21 group were significantly higher than those in LV-anti-miR-21 group.The early apoptotic rate and the inhibitory rate of cetuximab for the cells in LV-anti-miR-21 group were higher than those in LV-miR-21 group.CONCLUSION: miR-21 promotes the proliferation of colon cancer cells.Down-regulation of miR-21 enhances the sensitivity of the colon cancer cells to the targeted therapy drug cetuximab.