Objective To investigate the effects of c-myc promoter binding protein(MBP-1)gene expression silencing on the pro-liferation in vitro in human gastric cancer cell line SGC-7901.Methods The cells divided into three groups:blank control group (cells without transfecting gastric cancer cell),negative control group(cells transfecting missense sequence)and experimental group (cells transfecting MBP-1 shRNA).Two MBP-1 shRNA sequences and one negative control shRNA sequence were designed,syn-thesized and cloned into pSIREN-retroQ plasma.Then the recombinant plasmids were constructed and transfected into human gas-tric cancer SGC-7901 cells by Lipofectamine 2000.After selection of cells with puromycin for two weeks,stable cell line SGC-7901 expressing gene of MBP-1 specific shRNA was obtained.The expressions of MBP-1 mRNA and protein in SGC-7901 were deter-mined by the real time PCR and Western blot,respectively.The effects of altered expression of MBP-1 on the cell proliferation were measured by MTT cell proliferation assay.Results PCR and sequencing indicated that the recombinant plasmids pSIREN-retroQ was constructed.Then the recombinant plasmids were transfected into human gastric cancer SGC-7901 cells by Lipofectamine 2000. After selection of cells with puromycin for two weeks,stable cell line SGC-7901 expressing gene of MBP-1 specific shRNA was ob-tained.The relative expression level MBP-1 mRNA in the MBP-1 siRNA transfection group was significantly decreased compared with the blank control group(P <0.05).Compared with the blank group,the expression levels of MBP-1 protein in the experimental group also significantly decreased.The proliferation abilities of SGC-7901 cells at 48,72,96,120 h after MBP-1 siRNA transfection were significantly increased compared with the blank control group (P < 0.05 ).Conclusion Down-regulating the expression of MBP-1 can obviously promote the proliferation of human gastric cancer cell line SGC-7901.MBP-1 gene may become the new target of gene therapy for gastric cancer.

Establishment and improvement of rules and regulations will benefit to the management of research institutions.A comprehensive,fair,reasonable,and workable rules and regulations are the basic guarantee for maintaining the orderly,steady,and sustainable development of the institutions.It is also help to fully mobilize the enthusiasm and creativity of all members of the institutes.In this paper,we use an example of a construction of rules and regulations in the military medical research Institutions,to analyze current information and posted suggestions on the rules and regulations formulation principles,making procedure,overall framework and main innovation for the future improvement and implementation of rules and regulations.

Objective To study the mechanism of adaptive immunity against Rhizopus arrihizus (R. arrihizus) infections. Methods Bone marrow derived dendritic cells (BMDCs) were separated from C57BL/6 mice and Card9-/- mice and then were cultured in vitro. Resting spores and swollen spores of R. arrihizus were in vitro co-cultured with BMDCs with or without Syk inhibition. Secretion of cytokines ( IL-23, IL-1βand IL-12) was analyzed by ELISA after 24 hours of culture. Na?ve T cells derived from C57BL/6 mice were in vitro co-cultured with spore-stimulated BMDCs for four days. Levels of IL-17A and IFN-γ in supernatants of cell culture were analyzed by ELISA. Flow cytometry was performed to analyze T cell differ-entiation. Confocal microscopy was used to observe the images of stained β-glucan on the surface of resting and swollen spores. Swollen spores were co-cultured with Dectin-1, Dectin-2, TLR2 and mannose receptor ( MMR) , and the binding results were analyzed by flow cytometry. Results Swollen spores but resting spores could induce the maturation of BMDCs and promote the secretion of cytokines (IL-23, IL-1βand IL-12). Co-culturing T cells with swollen spore-stimulated BMDCs enhanced their differentiation to Th17 and Th1. In addition, swollen spores promoted the secretion of Th1-related cytokine ( IFN-γ) and Th17-related cytokine (IL-17A). Adding Syk inhibitor to Card9-/-BMDCs or wild type BMDCs significantly inhibited the secretion of cytokines and T cell differentiation, especially in the Card9-/- group. β-glucan was overserved on the surface of swollen spores, but not on resting spores. On the surface of swollen spores existed pathogen associated molecular patterns ( PAMPs) that could bind with Dectin-1 and TLR2. Conclusion Swollen spores of R. arrihizus could active BMDCs to secrete cytokines of IL-23, IL-1β and IL-12 and trigger T cell responses in vitro. The possible mechanism might be associated with β-glucan exposed on the surface of swollen spores that binds with Dectin-1. The responses between BMDCs and R. arrihizus are Syk-Card9-dependent.

Objective To investigate the relationship between heart rate variability and type 2 diabetes mellitus(T2DM)in Kirgiz and Han,to understand the differences between ethnic groups,at the same time to provide effective method for type 2 diabetic autonomic neuropathy.Methods 24 -h Holter recording was performed in 1 46 individuals.75 cases of T2DMconfirmed by glycolated hemoglobin(HbA1 c)were selected as observation group, Kirgiz 40 people,Han 35 people.While 71 subjects without T2DM were chosen as control group,Kirgiz 34 people, Han 37 people.The HRV indices,including standard deviation (SD)for the time between normal -to -normal complexes in the entire 24 h electrocardiographic recording(SDNN),the average of SD for the time between normal -to -normal intervals for each 5 min period(SDNN5),the percentage of adjacent RR intervals 50 ms apart(PNN50), normal RR intervals divided by RR intervals derived from the triangle index(TRIA),SD of the average normal -to -normal intervals for each 5 min period (SDANN)and root -mean square of differences of successive RR intervals (RMSSD)were compared between these groups.Results There were no significant differences in age,gender, education level and smoking between Kirgiz and Han patients.The differences were statistically significant in SDNN, SDNN5,PNN50,SDANN,TRIA(t =3.392,P =0.001 ;t =3.652,P =0.001 ;t =3.826,P =0.001 ;t =3.235,P =0.002;t =2.378,P =0.02)between patients with or without T2DMin Kirgiz population.In contrast,the mean square root of SDNN,SDNN5,PNN50,TRIA,RMSSD(t =3.498,P =0.001 ;t =3.41 4,P =0.001 ;t =2.1 05,P =0.039;t =2.1 02,P =0.039;t =3.41 3,P =0.001 )were statistically significant in Han population between T2DM patients and nondiabetic patients.There was no deference between Han and Kirgiz patients without T2DM in HRV,whereas the TRIA(t =2.887,P =0.005 )had significant difference in T2DM patients between Han and Kirgiz population. Conclusion The HRV is significantly reduced in the T2DMpatients,especially the Kirgiz.The HRV analysis using 24h Holter is a noninvasive technology to detect the T2DM.

Objective To study the effects of macrophage migration inhibitory factor (MIF) on spinal cord injury. Methods C57BL/6 mice with injury at T9 ~T10 were taken as spinal cord injury ( SCI) model, and they were divided into the operated group and the sham oper-ated group. Immunohistochemistry was used to detect expression changes of MIF during 72 hours after operation. Reverse transcriptase-poly-merase chain reaction ( RT-PCR) was used to test changes of mRNA level. And double staining immunofluorescence techniques was used to show expressions of MIF and RhoA. Results Expression of MIF at acute stage (72 hours after injury) increased significantly. Results of RT-PCR showed that mRNA levels of MIF and RhoA both increased. There were statistical differences between the two groups (P<0. 05). Con-clusion In acute stage after SCI, MIF increased in the activated microglia. MIF accumulated in oligodendrocyte-like and astrocyte-like cells in injured spinal cord, which might contribute to inhibitory environment for regeneration. Moreover, mRNA level of MIF raised with the in-crease of RhoA mRNA level, which indicated the potential inhibition of MIF to axonal regeneration in CNS.

Objective To study the curative effect of minimally invasive surgery in the treatment of rectal cancer.Methods 100 patients with rectal cancer were researched.They were randomly divided into observation group and control group,50 cases in each group.The control group received traditional abdominal radical operation,the observation group was treated with minimally invasive surgery.The operation effect was compared between the two groups.Results The operation time,length of stay and feeding time after operation in the observation group were shorter than those in the control group[(78.45±12.34)min vs.(98.24±25.31)min,(3.12±0.43)d vs.(7.53±1.12)d,(1.23±0.30)d vs.(3.56±0.57)d](t=4.967,25.992,25.578,all P<0.05).The amount of bleeding during operation of the observation group was less than that of the control group[(11.23±2.31)mL vs.(26.58±4.04)mL](t=23.323,P<0.05).The incidence rate of postoperative complications of the observation group was significantly lower than that of the control group[6.00%(3/40) vs.20.00%(10/50)](x2=4.332,P<0.05).After surgery,the quality of life in the observation group was higher than that in the control group[(57.82±6.84)points vs.(48.23±4.50)points](t=8.282,P<0.05).After 1 year of follow-up,there were no significant differences in metastasis rate,recurrence rate and survival rate between the two groups[2.00%(1/50) vs.6.00%(3/50),0.00%(0/50) vs.4.00%(2/50),98.00%(49/50) vs.92.00%(46/50)](x2=1.042,2.041,1.895,all P>0.05).Conclusion The effect of minimally invasive surgery for patients with rectal cancer is significant,it is worthy of promoting.

BACKGROUND: With the prevalence of total knee arthroplasty, periprosthetic joint infection as a serious complication has been an issue of concern.OBJECTIVE: To summarize the advances in the diagnosis and treatment of infection after total knee arthroplasty.METHODS: The first author searched the PubMed and CNKI databases for the literatures related to the diagnosis and treatment of infection after total knee arthroplasty using the keywords of total knee arthroplasty,infection in English and Chinese, respectively. The repetitive articles were excluded, and the risk factors,diagnosis, classification and treatment of infection after total knee arthroplasty were summarized.RESULTS AND CONCLUSION: (1) The infection after artificial knee replacement is closely related to the patient's preoperative condition, operation skills and postoperative status. (2) Diagnosis of infection is based on the detailed understanding of the patients' situation and complete auxiliary examinations. (3) The treatment methods include antibiotics, debridement and retention, reversion, arthrodesis, as well as amputation. (4) Periprosthetic infection is a serious complication after total knee arthroplasty, and early diagnosis and treatment are critical. Furthermore,choosing an appropriate individualized treatment scheme for different patients is important.

Aim To investigate the effects of Ang Ⅱ receptor antagonist irbesartan on the expressions of connective tissue growth factor(CTGF) and membrane-type 1 matrix metalloproteinase(MT1-MMP) in high glucose-cultured rat glomerular mesangial cells(GMCs).Methods High concentration glucose and irbesartan were used to stimulate the cultured rat GMCs in vitro.The mRNA and protein expressions of CTGF and MT1-MMP were detected with semi-quantitative RT-PCR and Western blot.The secreted collagen Ⅳ in the supernatants of the GMCs was detected by enzyme-linked immunoadsorbent assay(ELISA).Results Compared with control group,the expressions of CTGF were continuously increased in GMCs under high concentration glucose medium;otherwise the mRNA and protein levels of MT1-MMP in GMCs were decreased in a time-dependent manner at the same time.These changes were accompanied by increased secretion of collagen Ⅳ.Irbesartan could inhibit those changes induced by high glucose.Conclusions High glucosecould induce the expression of CTGF and inhibit the expression of MT1-MMP in GMCs.Irbesartan could inhibit the secretion of ECM in GMCs under high concentration glucose medium,partly by regulating the expressions of CTGF and MT1-MMP.

Objective To assess the mortality,complications and major morbidity of pneumonectomy for non -small cell lung cancer(NSCLC)and to establish the importance of various prognostic factors.Methods The data of 64 consecutive patients who underwent pneumonectomy for NSCLC were retrospectively reviewed.Results The 30 -day mortality rate was 7.8%(5 /64).Complications developed in 29.7%(1 9 /64)and overall 5 -year survival was 1 9.0%.Pathological stage Ⅲ(P =0.030)and right pneumonectomy(P =0.01 0)were independent risk factors of an adverse outcome.Survival was not significantly influenced by histological types (P =0.1 25)or curability (P =0.587).Conclusion Pneumonectomy is associated with acceptable overall morbidity and mortality.However,the patients with pathological stage Ⅲ or right pneumonectomy require special consideration.Pneumonectomy should be performed only in selected patients.

Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.