Objective To construct an eukaryotic expression vector encoding an shRNA targeting CA916798. Methods According to the CA916798 cDNA sequence in GenBank, 2 pairs of oligo nucleotides were designed and synthesized. After primer annealing, they were inserted into plasmid pGenSil-l to construct the shRNA eukaryotic expression vector. The recombinant plasmid were transformed into DH5?, and the positive strain were identified by enzyme digestion and sequence analysis. The recombinant vector were transfected into A549/CDDP cells with Lipofectamine 2000. Drug sensitivity and proliferation of the transfected cells were measured by MTT test. Results The pRNAi-CA916798 shRNA of recombinant plasmid was constructed successfully. The growth of A549/CDDP cells transfected by pRNAi-CA916798 shRNA was slowly than that of those with blank vector transfection after 1.0 ?g/ml CDDP treatment (P

Objective To explore the influence of low dose cyclophosphamide (CTX)acting on regu-latory T cells (Tregs)of hepatocellular carcinoma-bearing mice,and the anti-tumor effect of low dose CTX combined with cytokine induced killer cells (CIKs).Methods Models of tumor-bearing mice were established by subcutaneous inoculation with hepatocellular carcinoma cells.The mice were randomly divided into 4 groups (n =35),there were normal control group,single tumor group,single CTX group (1 time,1 00 mg/kg,intra-peritoneal)and cyclical CTX group (once every 6 days,3 times,1 00 mg/kg,intraperitoneal).The changes of the Treg/CD4 + in spleens were detected by flow cytometry.We isolated mononulear cells from spleen of mice to culture CIKs.To study the effect of anti-tumor in vitro,tumor-bearing mice were randomly divided into 6 treat-ment groups,there were single tumor group,single CTX group,single CIK group,cyclical CTX group,single CTX +CIK group and cyclical CTX +CIK group.The growth curves of tumor were drawn.At the end of the experiment,tumor was separated and weighted.The growth inhibition ratio of tumor was calculated.Results With time prolonged after tumor inoculation,the proportion of Treg/CD4 + increased gradually in spleen of mice in single tumor group.On the 1 0th day,this proportion of single CTX group was (8.95 ±1 .90)% and the sin-gle tumor group was (9.25 ±1 .74)%,and there was no difference between two groups (t =0.374,P =0.71 4).The proportion in the cyclical CTX group (8.99 ±2.1 1 )% was still lower than that in the single tumor group (1 6.76 ±2.02)% on the 1 9th day,and the difference was significant (t =8.544,P =0.000). Treatment for 21 days,the volume and weight of the single tumor group,single CTX group,single CIK group, cyclical CTX group,single CTX +CIK group and cyclical CTX +CIK group were (3 800.4 ±607.5 ), (3 764.8 ±537.7),(3 352.2 ±485.4),(2 076.6 ±620.2),(1 867.1 ±533.6),(970.7 ±1 35.3)mm3 and (4.01 ±0.66),(3.86 ±0.74),(3.80 ±0.42),(2.08 ±0.27),(1 .83 ±0.93),(0.86 ±0.25)g. The tumor volume and weight were minimum in the cyclical CTX +CIK group.The tumor inhibition effects were weaker in the cyclical CTX and single CTX +CIK groups.But tumor in single CIK and CTX groups were unsup-pressed.The differences among the 6 groups had statistically significance (F =21 3.750,P =0.000;F =27.1 42,P =0.000).Conclusion Cyclical administration of low-dose CTX combined with CIKs has an obvi-ous therapeutic effect on hepatocellular carcinoma bearing mice and can provide a new idea for anti-tumor therapy.

Objective To study the application effect of clinical pathway teaching in respiratory medicine. Methods Seventy clinical medical students of our department during 2007 to 2009 were randomly divided into two groups. The control group (35 students) was treated by traditional teaching ways, while the experimental group(35 students) was treated by clinical pathway teaching ways, with 5 to 6 students forming a small group. Teachers provided one copy of the CP version to each person in advance. Then progressive questions and discussions were conducted according to the diagnosis, dif-ferential diagnosis and treatment of CP. After its implementation for a certain time, students were co-mprehensively assessed by the practice examination and questionnaire and the statistical analysis was performed by using SPSS 17.0 version statistics software. Results The experimental group's total aver-age score was (90.00±4.00) points, while the control group's total average score was (76.00±7.20) points. There was significant difference between the two groups(P=0.001). The effect of these two kinds of teaching was evaluated and compared in stimulating interest in learning (P=0.002), improving the analytical ability(P=0.004),improving self-study ability(P=0.001), deepening the under-standing of the basic concepts(P=0.112), improving the innovation ability (P=0.005), improving the efficiency of learning(P=0.034), improving clinical comprehensive ability(P=0.016), and improving the ability of language expression(P=0.000), showing that teaching method of clinical pathway could significantly improve clinical teaching effect, and there was statistically significant difference between them. Conclusion Clinical pathway teaching has obvious advantages in cultivating students' diagnostic thinking and clin-ical ability to solve practical problems, and therefore it has better clinical teaching effect than the tra-ditional teaching method.

BACKGROUND:In previous experiments, TiO2-xNy OBJECTIVE:To study the friction properties of orthodontic brackets coated with TiO-coated bracket has been confirmed to have excelent antibacterial properties and biological safety performance. 2-xNy METHODS: TiO film. 2-xNy film was prepared by radiofrequency magnetron sputtering on MBT bracket (0.022″). The TiO 2-x N y RESULTS AND CONCLUSION: TiO-coated brackets were analyzed by X-ray diffraction and scanning electron microscopy observations. The coefficient of static friction and coefficient of kinetic friction between the wires (0.012″, 0.014″, 0.016″) and orthodontic brackets were calculated. 2-xNy film on the bracket was of anatase structure, which was compact and had good crystalinity. Under dry condition, the coefficient of static friction and kinetic friction of the brackets coated with TiO2-xNy were less than those of ordinary brackets; under wet condition, the coefficients of static friction and kinetic friction of the brackets coated with TiO2-xNy were less than those of ordinary brackets, but the difference was not statisticaly significant. Nano-TiO2-xNy film can reduce the friction between bracket and archwire, which wil offer a novel opportunity to significantly reduce the friction during tooth movement.

Meisoindigo is an indigo natural derivative commonly used in anti-cancer therapy. In the clinical application, it was also found to have good therapeutic effect on psoriasis. In order to further understand its mechanism of action, human normal keratinocyte cell line HaCaT and RAW 264.7 were used to identify if meisoindigo could affect the inflammatory factors such as NO and other important cytokines which were highly involved in psoriasis. Our results indicated that meisoindigo decreased the production of NO in LPS-stimulated RAW 264.7 cells and reduced the expression of cytokines in LPS-stimulated HaCaT cells. And TLR4-TAK-NF-kappaB was a possible pathway mainly involved in the attenuation of iNOS and pro-inflammatory cytokine production by meisoindigo, which may take part in the therapeutic effect of meisoindigo on psoriasis.

BACKGROUND:With more and more elderly patients suffering lumbar degenerative disease undergoing internal fixation, infection after spinal internal fixation is a common complication in orthopedic surgeries, but its treatment strategy remains controversial.OBJECTIVE: To explore the curative efficacy of incision infection after internal fixation in the elderly with lumbar degenerative disease.METHODS: 197 patients with lumbar degenerative disease undergoing internal fixation and followed up for more than 2 years admitted in Department of Orthopedics, Beijing Shijitan Hospital Affiliated to Capital Medical University from January 2012 to January 2015, were analyzed retrospectively. The follow-up time was 2-4.9 years. There were 97 cases of lumbar stenosis, 29 cases of lumbar disc herniation, 33 cases of lumbar spondylolisthesis, 17 cases of degenerative scoliosis and 21 cases of lumbar compression fractures.RESULTS AND CONCLUSION: (1) Eleven patients experienced incision infection, including ten acute, and one delayed infection. (2) Among acute infected cases, three were superficial infection in three cases and seven had deep infection,who characterized as exudation (10/10), local pain (8/10) and fever (9/10). Acute infected cases received bacterial culture, drug sensitive test, antibiotic therapy, and debridement of the infected wound, and leaving all internal fixators in situ in all but one case. (3) For delayed infection, one patient had local pain, incision exudation, and intermittent fever,and then the internal fixators were removed. (4) Pseudarthrosis was not founded during 2-year follow-up in all patients.(5) These results suggest that for the elderly patients suffering lumbar degenerative disease with infection after internal fixation, intravenous antibiotics, debridement plus drainage are recommended, but without internal fixator removal, and repetitive debridement and drainage is a rational choice if necessary.

Syl948 is a synthesized selective S1P1 agonist with novel structure. HTRF-IP1 test indicated that Syl948-P, the active form of Syl948 in vitro, has strong activity against S1P1 (EC50: 83 +/- 16 nmol x L(-1)), but its effect on S1P3 was very weak (EC50: 1 026 +/- 90 nmol x L(-1)). In SD rats, oral administration of Syl948 10 mg x kg(-1) significantly decreased the peripheral blood lymphocytes (PBL), with the maximal PBL inhibition rate of 63%, which was as similar as equal dose of fingolimod (FTY720). Oral administration of Syl948 10 mg x kg(-1) had no effect on heart rate of SD rats, which was better than FTY720. Daily oral administration with Syl948 (2 or 4 mg x kg(-1)) significantly prolonged the survival time of the allografts of skin slice on mice. In summary, the above results demonstrated that Syl948 has great selectivity in vitro and good activity in vivo, which indicated its potential use as an anti-rejection drug in skin transplantation.

Objective To investigate the value of magnetic resonance imaging (MRI)-based intravenous thrombolysis in patients with wake-up ischemic strokes (WUIS).Methods Patients presenting within 12 hours of acute stroke symptom onset and those with WUIS confirmed by CT,excluding intracranial hemorrhage,were encouraged to perform an emergent brain MRI scan to confirm the diagnosis of hyperacute ischemic stroke (hyper-intense in DWI without hyper-intense change in T2WI or fluid attenuated inversion recovery (FLAIR)).These patients then received intravenous thrombolytic therapy with recombinant tissue plasminogen activator (rt-PA).All patients were divided into either stroke presenting within 12 hours or WUIS.The clinical outcomes were assessed by the modified Rankin scale (mRS) and the Barthal index (BI) at baseline and at 90 days after the thrombolysis therapy.Results Two hundred and sixty-one patients (261/563,56.4％) had confirmed diagnosis of hyperacute ischemic stroke (WUIS,n =73,73/121 =60.3％ vs within 12 hours,n =188,188/342 =55.0％).Altogether,192 patients (139 in within 12 hours group,and 53 in WUIS group) received intravenous thrombolytic therapy with rt-PA.No significant differences were found between the 2 groups at the baseline characteristics and at 90 days outcomes after the thrombolysis therapy(x2 =1.296 and 1.473,P =0.538 and 0.489,respectively).Also no significant differences were found in the incidence rate of secondary hemorrhage (including both of asymptomatic and symptomatic) and mortality rate between the 2 groups.Conclusion MRI-based intravenous thrombolysis is safe and effective in the treatment of patients with hyperacute WUIS.

Aim To determine the effect of SYL934 , a novel immunosuppressant, on skin allograft rejec-tion. Methods HTRF-IP1 assay was used to evaluate the agonistic activity of SYL934-P, the active form of SYL934 in vivo, on S1P1 and S1P3 in vitro. SD rat peripheral blood lymphocytes ( PBL ) test and heart rate test was used to assess the in vivo immunosuppres-sive effect and heart rate effect of SYL934 . Mice skin graft transplantation experiment was used to observe the effect of SYL934 on skin allograft refection. ResultsSYL934-P selectively activated S1 P1 but not S1 P3 in vitro. Single orally administration of SD rats with
SYL934 decreased the PBL significantly and played an obviously immunosuppressant role, but did not affect the heart rate. Daily orally administration of mice with SYL934 significantly increased the survival rate of al-lografts of skin slice in mice. Conclusion SYL934 has great selectivity in vitro and good activity in vivo, which indicated it potential use as an anti-rejection drug in skin transplantation.

Objective To investigate the serum levels of 25-hydroxyvitamin D3 (25 (OH)D3) and urine vitamin D binding protein(uVDBP) in patients with diabetic nephropathy (DN),and to determine the relationship between 25 (OH) D3,uVDBP and DN,in order to provide a new method for early diagnosis and treatment of DN.Methods From January 2015 to December 2015,85 DN patients admitted into Weihai Municipal Hospital were selected.According to the ratio of UALB to UCR(UACR),the patients were divided into three groups.Type 2 diabetes had 28 cases of normal albuminuria group,31 cases of microalbuminuria group,and 26 cases of clinical albuminuria group.We also enrolled 25 healthy people who received outpatient service as control group.Serum 25 (OH) D3 levels were measured by chemiluminescence method.Urine VDBP levels were assayed by ELISA.FPG,HbA1 c,UREA,SCr,TC,TG were measured by electrochemiluminescence.Results The results showed that serum 25 (OH)D3 was significantly lower in the normal albuminuria group,microalbuminuria group and clinical proteinuria group than that in the control group (P ＜ 0.05),and there was statistically significant difference among the four groups [(20.04 ± 7.52) ng/mL,(16.54 ± 6.51) ng/mL,(10.77 ± 4.63) ng/mL,(29.65 ± 5.47) ng/mL,F =86.294,P ＜ 0.001].The results showed that uVDBP was significantly higher in the DN group than that in the control group(all P ＜ 0.05),and there was statistically significant difference among the four groups [(8.44 ± 3.20) mg/L,(14.22 ± 3.26) mg/L,(2 1.77 ± 5.87) mg/L,(4.95 ± 1.34) mg/L,F =125.583,P ＜ 0.001].Correlation analysis showed that serum 25 (OH) D3 decreased gradually with the increase of DN and negatively correlated with UACR (r =-0.575,P ＜ 0.01),while uVDBP level was positively correlated with UACR (r =0.436,P =0.015).Conclusion With the progress of DN,serum 25 (OH) D3 levels gradually decreased,indicating that 25 (OH) D3 may play an important role in the pathogenesis of DN;uVDBP may be an early diagnostic method for DN.