Natural killer (NK) cells play an important role in the immune response to viral infection and tumors.The ability of NK cells depends on the integrated signals across the array of stimulatory and inhibitory receptors engaged upon interaction with target cell surface ligands. Some stimulators, including viruses, tumor cells and hot shock, could promote the expression of NKG2 receptors and their ligands via activating certain transcription factors which are capable of up-regulating NKG2 promoters' activity. Meanwhile, epigenetic mechanisms including DNA methylation and histone posttranslational modifications are also critical to expression of NKG2 receptors and their ligands and may control the clonally distribution of some NK cell receptors. Investigating the epigenetic mechanisms of NK cell receptors and their ligands might helpful to the rational design of therapy against infection, inflammation, cancer or autoimmune diseases.

Objective To investigate the effects of nuclear factor κB(NF-κB) antisense oligodeoxynucleotides (AS-ODNs) on transforming growth factor β1 (TGF-β1)-induced epithelial mesenchymal transition (EMT) in human renal tubular epithelial cells. Methods NF-κB AS-ODNs were transferred into the human renal tubular epithelial cells (HK-2), and the cells were stimulated by 10 μg/L TGF-β1 for 24 hours. The expression of NF-κB mRNA and α-SMA mRNA were measured by RT-PCR. α-SMA protein expression was assessed by fluorescence spectrum.Results TGF-β1 significantly up-regulated the expression of NF-κB mRNA, which was 8 folds of blank control (P<0.01). TGF-β1-indueed epithelial mesenchymal transition was inhibited by NF-kB AS-ODN and the NF-KB mRNA expression of AS-ODNs was decreased by 75%(P<0.05).The expression of α-SMA mRNA and protein was also down-regulated obviously (P<0.05).Conclusion NF-κB AS-ODN can inhibit the expression of NF-κB and the epithelial-mesenchymal transition, which may be a new therapeutic strategy against tubulointerstitial fibrosis.

Objective To explore the nursing measures of patients with pure diffuse axonal injury (DAI)treated by multi-step sub-hypothermia therapy.Methods The nursing care of pure DAI patients treated by multi-step sub-hypothermia therapy,which included the traumatic condition evaluation before treatment,the nursing care during treatment,the nursing care after treatment but also in coma,and the nursing care after palinesthesia.Results The intracranial pressure and concentration and lactic acid in blood and cerebrospinal fluid alleviated.The forehead expanded.The disability rating scale(DRS)decreased.While the incidence of sub-hypothermia related complications did not increased.Conclusions The elaborative nursing care aiming at different pathogenetic conditions,different stages is the first guarantee in the treatment of pure DAI with multi-step sub-hypothermia therapy.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced podocyte injury and its signal transduction mechanism.Methods Differentiated mouse podocytes were exposed to normal glucose,high glucose,and different concentrations of 1,25(OH)2D3 or LY294002 (a selective PI3K inhibitor) for 24 h.PCR and immunofluorescent staining were used to detect nephrin,podocin,and desmin.Western blotting was used to detect protein expression of nephrin,podocin,desmin,PI3K,Akt and p-Akt.Results Compared with high glucose group,1,25(OH)2D3 (100 nmol/L and 1000 nmol/L) significantly up-regulated the expression of podocin and nephrin in podocytes induced by high glucose (P ＜ 0.05).Meanwhile,1,25(OH)2D3 (100 nmol/L) significantly reduced the expression of desmin (P ＜ 0.05).PI3K and p-Akt were obviously reduced in high glucose group.In the presence of 1,25(OH)2D3,the trends were reversed.However the above effects of 1,25(OH)2D3 were abolished when p-Akt was blocked by the PI3K inhibitor LY294002.Conclusions 1,25 (OH)2D3 can inhibit high glucose-induced pedocyte injury through PI3K/p-Akt signaling pathway.

Objective To develop a kind of new machine for making animal model for pressure ulcer,and inspect its effect through experiments,in order to lay the foundation for the research of pressure ulcer experiments on animals.Methods This study developed the machine after reviewing the domestic and foreign literature,making full use of the existing experimental platform of our university.Then 55 Sprague Dawley(SD) rats were selected,after anesthesia and the skin preparation,the researchers imposed certain pressure with 70 mmHg/cm2 (1 mmHg=0.133 kPa) on the skin and muscle tissue on the inner left thigh of SD rats by using self-designed machine,pressing for 2 h,then reperfusion for 30 min,3 times a day,a total of 7 days.Results The authors developed a kind of new machine for making animal model for pressure ulcer,and successfully prepared Ⅲ phase pressure ulcers model in SD rats with success rate of 98.2％(54/55).Conclusion This machine can prepare Ⅲ phase pressure ulcers model on animals,it's easy to use and efficient,it can be used for researches in the field of prevention and cure of pressure ulcers.

Objective To investigate the effect of active vitamin D (VD) on macrophage M1 and M2 phenotype and its role in protecting podocyte impairment in diabetic nephropathy (DN).Methods Diabetes mellitus rats were established by intraperitoneal injection with streptozocin.Rats were randomly divided into four groups:normal-1 (NC-1,n=8),normal-2 (NC-2,n=8,normal rats treated with calcitriol 0.1 μg· kg-1 · d-1 by gavages),DN (n=24) and VD (n=24,DN+calcitriol 0.1 μg· kg-1 · d-1 by gavages).Blood glucose and body weight were assessed,and 24-hour urine was collected regularly.Blood and urine samples were taken for biochemical study,and kidney tissues were used for PAS staining to assess histological changes.Immunohistochemical staining was used to detect number of CD68 + macrophage.Western blotting was used to detect protein expressions of nephrin,podocin,CD68,M1 specific marker of inducible nitric oxide synthase (iNOS),TNF-α and M2 specific marker of CD163,arginase 1 (Arg-1),mannose receptor (MR).Results (1) In DN group,levels of BUN,Scr,urinary protein and glomerular mesangial matrix proliferation were significantly higher (P ＜ 0.05),and the expressions of nephrin,podocin were significantly decreased compared with NC groups (P ＜ 0.05).These above changes were significantly improved in VD group (P ＜ 0.05).(2)Number of CD68 + macrophage infiltration in DN group was increased in a time dependent manner compared with NC groups,which was significantly reduced in VD group (P ＜ 0.05).(3)To further definite M1 and M2 macrophage activation phenotype,the protein expressions of iNOS and TNF-α was increased in DN group at 8th,14th,18th weeks compared with NC groups (P ＜ 0.05),which were significantly decreased in VD group (P ＜ 0.05).Although,there were no significant difference of protein expressions of CD163,Arg-1 and MR between VD and DN group at both 8th and 14th week (P ＞ 0.05),the protein expressions of CD163,Arg-1 and MR were higher in VD group at 18th week than that in DN group (P ＜ 0.05),and the ratio of CD163/CD68 was also enhanced in VD group (P ＜0.05).(4)Moreover,the protein expression of iNOS was negatively correlated with expression of either nephrin or podocin (r =-0.707,P ＜ 0.01; r =-0.712,P ＜ 0.01),whereas the protein expression of CD163 was positively correlated with expression of either nephrin or podocin (r =0.627,P＜ 0.01; r=0.613,P ＜ 0.01).Conclusion Vitamin D can regulate macrophage phenotype,via inhibiting M 1 macrophage activation and enhancing M2 macrophage activation to protect podocyte impairment.

Objective To investigate the effects and underlying mechanism of calcitriol on ameliorating podocytes impairment in DN rats.Methods SD rats were randomly divided into four groups:normal control (NC) group,calcitriol treatment (VD) group:calcitriol 0.1μg· kg--1 d-1,diabetic nephropathy (DN) group:streptozocin (STZ) 58 mg/kg,DN treated with calcitriol (DN + VD) group:calcitriol 0.1 μg · kg-1 · d-1 + STZ 58 mg/kg.Rats were sacrificed at the end of 18 weeks.Results Compared with the DN group,the DN + VD group exhibited significantly lower proteinuria by 36％,improved renal histology at the end of the experiment (P ＜ 0.05),and similar levels of blood glucose,serum urea nitrogen as well as body weight (P ＞ 0.05).There were no significant differences in the serum concentrations of creatinine,calcium and phosphorus among the four groups (P ＞ 0.05).In DN group,the expressions of nephrin,podocin,VDR,PI3K-p85 and p-Akt were significantly decreased and the expression of desmin was increased compared to NC group.Calcitriol treatment could attenuate the above changes.Additionally,a positive correlation was observed between the expressions of nephrin and VDR (r=0.776,P ＜ 0.05).Likewise,the expression of nephrin was positively correlated with either PI3K -p85 or p-Akt (r=-0.736,r=0.855,all P ＜ 0.05).Conclusion Calcitriol can ameliorate podocytes injury in DN rats,which might be related with the further up-regulation of PI3K/p-Akt signaling pathway.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced macrophage activation and its underlying signal transduction mechanism.Methods RAW 264.7 cells were used to perform cell culture,the activity of intracellular iNOS was measured.VDR siRNA and PPARγ antagonist pre-treatment with macrophages were done before using 10-8 mol/L1,25(OH)2D3 to intervene high glucose pre-incubated macrophages.M1 markers including iNOS,TNF-α,IL-12,M2 markers including MR,Arg-1,IL-10 and nuclear receptors VDR and PPARγ were separately examined.Results The iNOS activity was increased in a glucose-dose and time dependent manner.Particularly,25 mmol/L glucose at 24 h gave the maximum response.After being treated with 25 mmol/L glucose for 24 h,not only inflammatory cytokines of TNF-α,IL-12 in the supernatant were increased,but quantitative real-time PCR and Western blotting analysis showed iNOS was also up-regulated (P ＜ 0.05).However,M2 markers,i.e.MR and Arg-l were significantly decreased (P ＜ 0.05).When in the presence of 1,25(OH),D3,the trends were reversed:the markers of M1,including TNF-α,IL-12 and iNOS were obviously reduced (P ＜ 0.05),while M2 markers,IL-10,Arg-1 and MR were increased (P ＜ 0.05).In addition,VDR and PPARγ were also increased (P ＜ 0.05).However,the above effects of 1,25 (OH)2D3 were abolished when further inhibited the expression of VDR and PPARγby VDR siRNA and PPARγ antagonist.Besides,accompanied by VDR,PPARγwas also decreased upon the treatment with VDR siRNA (P ＜ 0.05).Conclusion 1,25(OH)2D3 can promote high glucose induced classically activated macrophages (M1) converting to alternatively activated macrophages (M2) and this is achieved through VDR-PPARγ pathway.

Objective To investigate the sequence-dependent effects of gemcitabine plus erlotinib on the proliferation of human pancreatic carcinoma cells,BXPC-3 and PANC-1,and the possible mechanisms involved.Methods The expressions of mRNA and protein of endothelial growth factor receptor(EGFR)were determined by RT-PCR and Western blotting respectively.MTT assay was used to examine the effects of gemcitabine(3?10-11-3?10-2mol/L)and erlotinib(10-8-10-4mol/L),respectively,on the proliferation of human pancreatic carcinoma cells,then the IC50 was calculated subsequently.The effects of gemcitabine(IC50)and erlotinib(10-5mol/L)either administered alone,simultaneously,or sequentially(with 72h or 24h interval)on the proliferation of cells with MTT assay.Cell cycle was detected by flow cytometry.Results Both mRNA and protein of EGFR were expressed in BXPC-3 and PANC-1 cells.Gemcitabine(3?10-10-3?10-2mol/L)and erlotinib(10-6-10-4mol/L)significantly inhibited the proliferation of BXPC-3 and PANC-1 cells in a time-and concentration-dependent manner.The effects of gemcitabine plus erlotinib on cell proliferation were sequence-dependent.The inhibitory effects on cell proliferation was enhanced when administered simultaneously(P=0.034;P=0.049)or erlotinib was administered 24h prior to gemcitabine(P=0.001;P=0.025)in comparison to that of each drug used alone.However,administration of erlotinib 24h after that of gemcitabine(P=0.499,P=0.824)exerted no additive effects on the cell proliferation when compared with the effect of gemcitabine used alone.No statistical difference existed in the inhibitory effects on cell proliferation between the simultaneous administration of both drugs and the gemcitabine administration following erlotinib(P=0.199,P=0.767).Erlotinib plus/or gemcitabine treatment resulted in the block of cell cycle of BXPC-3 cells at G1 phase.Conclusions Both gemcitabine and erlotinib can inhibit the proliferation of BXPC-3 and PANC-1 cells.The concurrent administration or sequential administration of gemcitabine following erlotinib exerts stronger additive effects on cell proliferation than when gemcitabine is used alone.However,the additive effects are not related to the influence of the both drugs on cell cycles.

OBJECTIVE To investigate the expression of PTEN and VEGF gene in laryngeal squamous cell carcinoma and their clinical significance. METHODS The expression of PTEN and VEGF gene were examined by using immunohistochemical S-P staining method in 10 cases of normal tissues, 20 cases of para-tumor tissues and 60 cases of LSCC. RESULTS The positive rates of PTEN in normal tissues, para-tumor tissues and LSCC were 100.0 %, 95.0 %, and 70.0 % respectively (P＜0.05),and VEGF protein showed positive expression of 20.0 %,50.0 %,73.3% in normal, para-tumor and LSCC tissues with statistical significance(P＜0.05), In LSCC, the expression of PTEN and VEGF was positively correlated with TNM stage, differentiation of tumor, cervical and distant metastasis of lymph node, 3-year survive rate of patients(P＜0.05),No significant association was observed in expression of PTEN and VEGF with tumor location,patient's age and sex(P＞0.05). Spearman correlation analysis revealed that PTEN was negatively correlated with VEGF expression(?=-0.3948, P=0.0018). CONCLUSION The aberrant expression of PTEN and VEGF may play a role in occurrence, progression and metastasis of LSCC.