Objective To observe the effect of the joint treatment containing single sialic acid four hexose gangliosides,clopidogrel,atorvastatin in the treatment of cerebral infarction.Methods 88 acute cerebral infarction patients met the inclusion criteria were randomly divided into the observation group of 44 cases and the control group of 44 cases.The control group were given clopidogrel and atorvastatin,and the observation group were give single sialic acid four hexose gangliosides on the base of the control group,100mg/time,qd,4 week for a course.The symptoms before and after treatment were observed.The NIHSS and ADL score were calculated.Blood lipids,CRP,IMT was inspected.Results The total effective rate of the observation group was 90.91％,the control group was 77.27％,the observation group was significantly higher than the control group (x2 =11.967,P ＜ 0.05) ; the TC,TG,LDL-C,HDL-C,CRP,IMT,ADL,NIHSS of both group after treatment were significantly improved than before treatment,there were a statistically significant differences (all P ＜ 0.05).After treatment,the TC,TG,LDL-C,HDL-C,CRP,IMT,ADL NIHSS of observation group and the group were (4.06/ ± 0.98) mmol/L vs (4.18 t 1.02) mmol/L,(1.35 ±0.73)mmol/L vs (1.42 ±0.81)mmol/L,(1.68 ±0.82) mmol/L vs (1.74 ±0.85)mmol/L,(1.44 ±0.58) mmol/L vs (1.41 ±0.52)mmol/L,(40.55 ±3.12)mg/L vs (47.27 ±3.57)mg/L,(0.78 ±0.08)mm vs (0.87 ±0.09)mm,(28.9 ± 11.3) scores vs (22.3 ± 10.0) scores,(8.2 ± 5.3) scores vs (15.5 ± 6.2) scores.the CRP,IMT,ADL,NIHSS of observation group were improved more significantly than the control group,the differences were statistically significant(all P ＜ 0.05).Conclusion Jointing treatment containing single sialic acid four hexose gangliosides,clopidogrel,atorvastatin can improve significantly lipid levels,inhibit platelet aggregation,make a stability of carotid atherosclerotic plaque,improve activities of daily living and neurological deficit,but also can prevent secondary brain damage after traumatic brain injury,which is the ideal solution for the treatment of cerebral infarction.

Objective: To investigate the expression of recombinant human intedeukin-18 (hIL-18) in 3.7 L fermenter with the constructed engineer train Pichia pastoris X-33/hIL-18, and the procedures of expression and purification thereof.Methods: The train X-33/hIL-18 was inoculated in BMGY medium and then inoculated into the fermenter until the A600 of the culture reached about 6. The supernatant of fermentation was isolated and purified with centrifugal fiher devices, hydrophobic chromatography column and anion exchange chromatography column. Results: The recombinant hIL-18 was expressed in 3.7 L fermenter with batch feed methanol and the concentration reached 202 mg/L. After the purification, the purity could be more than 97%. The recombinant hIL-18 was shown to induce interferon-gamma (IFN-γ) production by human peripheral blood mononuclear cells (PBMCs), and enhanced NK cell cytotoxicity synergistically with IL-2. Conclusion: A great deal of the recombinant hIL-18 with higher purity could be harvested by Pichiapastoris expression system. This study showed a new potential for further study of its function and activities.

Objective To observe the effects of Yushen Heji on expressions of Heme Oxygenase (HO-1) and HO-2 in kidney of diabetic rats;To explore its mechanism in diabetic nephropathy. Methods SD rats were randomly divided into normal group, model group, control group and treatment group. Models were duplicated by subcutaneous injection STZ. Control group and treatment group were given gavage with related medicine, while normal group and model group were given gavage with the same amount of distilled water. Eight weeks later, the urine of rats was collected to detect the 24 h Upro and Ucr;the rats were sacrificed to detect the levels of CO, BUN and Scr;the kidneys were harvested for observing the expressions of HO-1 and HO-2 by immunohistochemistry.Results Compared with normal group, the contents of BUN, 24 h Upro and Scr all increased in model group;Ccr and plasma CO decreased;the expressions of HO-1 and HO-2 in kidney tissues were obviously reduced (P<0.01). Compared with model group, the contents of BUN, 24 h Upro and Scr all obviously decreased in treatment group and control group;Ccr and plasma CO increased;the expressions of HO-1 and HO-2 in kidney tissues obviously increased (P<0.05,P<0.01). Compared with control group, the contents of 24 h Upro decreased in treatment group;plasma CO increased;the expressions of HO-1 and HO-2 in kidney tissues obviously increased (P<0.05,P<0.01).Conclusion Yushen Heji has therapeutic effect on diabetic nephropathy rats, which may related with HO/CO system.

Fourteen H9N2 avian influenza viruses (AIV) were isolated from sick chickens in China from 1998 to 2008. The sequences of the Non-structural(NS) gene of these isolates were determined by RT-PCR and sequencing, and the entire ORF sequences of NS1 and NS2 protein were obtained.-The homology of these nucleotide sequences and the putative amino acid sequences were compared with several classic reference viruses of H9N2. These isolates were proved to be highly homologous in NS gene (92.9%-99.9% identity) and all belonged to A/Chicken/Beijing/1/1994-like group in the Asia bird-swine branch of allele A of HS gene phylogenetic tree.-According to this study and previous reports of other researchers, NS gene of H9N2 subtype AIV in chickens of China is genetically stable and there is no enough evidence to support the establishment of other sub-lineages in chickens.

Objective To study the effect of feto-matemal microchimerism in the treatment of activated human leukocyte antigen (HLA) haploidentical mobilized peripheral blood cells against solid tumors. Methods Genomic DNA samples of 25 pairs of HLA haploidentical donors and recipients were extracted. The donor-derived HLA-DRB loci were detected with nested PCR-sequence specific primer(SSP) typing. The mixed lymphocyte proliferation action between the patients and respective donors, the engraftment of donor's cells and the serum levels of Th1/Th2 type of cytokines were measured with MTT,FISH and EIJSA method respectively. The survival time of patients with or without feto-matemal microchimerism were compared as well. Results Using nested PCR-SSP typing, the positive rates of feto-maternal microchimerism in the 25 pairs of HLA haploidentical donors and recipients were 40% in the maternal/children pairs and 0 in the paternal/children pairs. The chimerism positive patients showed less proliferation activity when cocultured with respective donors as compared with unrelated ones (P=0.03).Only one chimerism positive patient experienced the engraft of donor's cell 3 months after treatment as the donor derived XX chromosome was identified with FISH. When the data of chimerism positive patients were deleted, the serum levels of IFNγ 1 month after treatment dropped dramatically from 171.4 (26. 3～258.4) ng/L to 29. 4(1.2～39.9)ng/L. The survival time in chimerism positive patients of the maternal/children pairs was significantly longer than that in chimerism negative patients, which was (31.2±4. 3) months and (11.1±3.3) months, respectively (P=0.036). Conclusion Feto-maternal microchimerism might induce anergy in the HLA haploidentical donors, favor the engraftment of donor's progenitors and maintenance of positive microenvironment and prolong the survival time.

OBJECTIVE:To provide reference for rational use of antibiotics to treat Pseudomonas aeruginosa (PA) infection in the clinic. METHODS:Resistant rate of PA in our hospital during 2011-2015 were analyzed retrospectively. Antibiotics use densi-ty(AUD)of 10 commonly used antibiotics were analyzed statistically,and the correlation of resistant rate with AUD was investi-gated by Spearman correlation analysis. RESULTS:One thousaud and eleven strains of PA were isolated in our hospital during 2011-2014,detection rate of PA always occupied the top 5 place. Top 3 antibiotics in the list of AUD were levofloxacin,ceftazi-dime,cefoperazone sodium and tazobactam sodium. AUD of piperacillin sodium and tazobactam sodium,levofloxacin,ciprofloxa-cin and meropenem were positively correlated with resistant rate of PA(r were 1.000,0.900,1.000,1.000,P<0.05). AUD of ce-foperazone sodium and tazobactam sodium were negatively correlated with resistant rate of PA(r=-0.900,P<0.05). AUD of imi-penem and cilastatin sodium,ceftazidime,gentamicin,aztreonam and amikacin had no correlation with resistant rate of PA(P>0.05). CONCLUSIONS:There is correlation between AUD of antibiotics and resistant rate of PA. It is of important significance to detect resistant rate of PA and the use of antibiotics regularly. Antibiotics should be selected cautiously in accordance with bacterial monitoring data,results of drug sensitivity tests,the amount and resistant rate of antibiotics,etc,in order to reduce resistant PA.

Objective: To evaluate the anti-tumor and side effects of activated-HLA haploidentical peripheral blood tem cells (haplo PBSCs) in the treatment of advanced refractory solid tumor patients. Methods: Forty-two patients with advanced refractory tumor, who were diagnosed in our hospital from Oct. 2004 to Oct. 2007, were enrolled in this study (all patients signed informed consent), including 12 with ovarian cancer, 9 with renal cancer, 8 with lung cancer, 8 with breast cancer, 2 with colon cancer, 2 with gastric cancer, and 1 with melanoma. The donors were healthy direct relatives of the patients; the donors' haplo-PBSCs were mobilized, collected, and activated by rhIL-2 in vitro. The clinical efficacy and side effects of haplo-PBSCs therapy were assessed by CT/PET-CT scanning, RESIST standard, KPS score, and clinical response rates, etc. Results: All 42 patients received one episode of haplo-PBSCs treatment. The progression-free survivals (PFS) were 6 months and the clinical beneficial rate (CR+PR+SD) was 73.8%. The beneficial rate of life quality was 76.2% and the KPS increased by 20 (0-30) points on average after haplo-PBSCs treatment. The patients with KIR unmatched in GVH direction had better outcomes than those with KIR matched or KIR unmatched in HVG direction (P<0.05), and the clinical beneficial rate, PFS and total beneficial rate were 94.1% vs 60.0%, (13.4±1.3) vs (8.0±0.9) months, and 89.5% vs 65.2%, respectively (all P<0.05). The donor/recipient relation as the mother/child had a better outcome than that as the father/child (P<0.05). Patients with renal cancer or ovarian cancer had better outcomes than those with other cancers, with clinical beneficial rates being 90.0% and 81.8%, respectively. Conclusion: Activated haplo-PBSCs therapy can induce non-specific anti-tumor effect, and improve the clinical symptom and life quality of advanced tumor patients.

Case recruitment of large-scale clinical trials should be strictly checked in quality and quantity for it is the key to clinical trial. This study discusses the main difficulties and countermeasures in the case recruitment of large sample, multi-center clinical trials according to the national research project "Myocardial Infarction Secondary Prevention Study in Traditional Chinese Medicine".

Objective: To investigate the effect of RetroNectin on CIKs cells and the related mechanisms. Methods: Peripheral blood mononuclear cells (PBMC) were collected from patients and divided into two groups: group Ⅰ and group Ⅱ. Samples in group Ⅰ were seeded into culture flask precoated with RetreNec-tin and CD3mAb to induce CIKs. While samples in group Ⅱ were seeded into common culture flask. The pro-liferation of CIKs was detected by cytometric analysis. The cytotoxic activity of CIKs was determined by LDH assays. The phenotype changes and cell cycle of CIKs were identified by flow cytometry. The apoptosis of cells was detected by Annexin V/PI. Western blot was employed to detect the level of protein Vav1. The CD49d and CD49e were blocked by anti-CD49d and anti-CD49e and the proliferation of cells was tested by cytometric analysis after the blockage. The phenotype changes of cells were identified by flow cytometry after the blockage. Results: RetroNectin enhanced the proliferation of CIKs (P<0.05). Flow cytometric analysis showed that RetroNectin significantly increased the number of CD25+ T cells (P<0.05). RN-CIK was more ac-tive than CIK in killing HCT-8 cell lines in vitro (P<0.05). RetroNectin could block the CIKs at G_1 phase (P<0.05) and resist apoptosis. There was no significant difference in the proliferation between the two groups af-ter the blockage with CD49d and CD49e (P>0.05). The expression of protein Vavl was associated with CD25+T cells. Conclusion: RetroNectin enhances the proliferation of CIKs by influencing the cell cycle, resist-ing apoptosis possibly through the site of CD49d and CD49e, and inducing T cell activation as the second sig-naling through Vav1.