The elements in blood or serum, including calcium, magnesium, copper, zinc, chromium, manganese, nickel, selenium, lead and cadmium were determined in the normal and patients in Nanjing by atomic absorption spectrophotometry. The method used was proved to be reliable being checked by the reference bovine serum and the data obtained in this paper was coincident with those reported by the other authors previously. The relationship between the concentrations of elements and the ages as well as the sexes in the normal was discussed briefly. The concentrations of several of the elements of the patients were significantly different from those of the normal in the same ages.

Objective:To investigate the antitumor efficiency of the special cytotoxic T lymphocytes(CTLs) activated by dendritic cells(DCs) pulsed with K-ras (12-Val) antigen.Methods:DCs was generated from PBMC in the presence of granuloceyte/macrophage-colony stimulating factor(GM-CSF),interleukin-4(IL-4)in vitro.DCs were differently sensitized with K-ras mutant pancreatic cancer cell line,K-ras(12-Val) mutant peptide,K-ras(12-Val) mutant peptide with the surface of cationic nanoparticle.Cell surface markers on DCs was measured by flow cytometry.The activation of CTL induced by DCs was detected by ~3H- thymidine incorporation test.The killing effects of CTL to pancreatic cancer was detected by ~(125)I-UdR release test. Production of IL-12 and IFN-γ by DCs and PBMC was detected by ELISA.Results:Compared with DCs pulsed with K-ras(12-Val) mutant peptide and K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle,DCs pulsed with whole tumor antigen could better induce CTLs killing activity(P＜0.05).The DCs with K-ras(12-Val) mutant peptide and K-ras mutant peptide with the surface of cationic nanoparticle could produce specific CTL killing activity aganist pancreatic cancer cell line Patu8988(K-ras+)(P＜0.05),but not SW1990(K-ras-)(P＞0.05). K-ras (12-Val) mutant peptide with the surface of cationic nanoparticle at lower concentrations can be effectively presenting on the surface of DCs than only K-ras (12-Val) mutant peptide.Conclusion:K-ras (12-Val) mutant peptide with cationic carrier can be effectively presenting and expression of DCs and induce CTL specific killing activity aganist pancreatic cancer cell lines with K-ras (12-Val) mutant peptide.

Objective To obserVe the effect of gyPenoside on liPid Peroxidation and hePatic lesion in rats with tyPe 2 diabetes mellitus and nonalcohol fatty liVer disease. Methods Totally,65 SPF male SD rats were randomly diVided into blank control grouP (grouP N),NAFLD model grouP (grouP NM),and NAFLD with T2DM model grouP. The NAFLD with T2DM model grouP was further diVided into three subgrouPs:JH grouP,Perfused with 1 g·kg-1 ·d-1 GPS;JL grouP,Perfused with 0. 5 g·kg-1 ·d-1 GPS;model control grouP,Perfused with the same Volume of water. Blood sugar,triglycerides ( TG) ,total cholesterol ( TC) ,alanine aminotransferase ( ALT) ,asPart aminotransferase ( AST) ,adePonectin ( ADP) in the Plasma were measured. TG, malondialdehyde (MDA),and suPeroxide dismutase (SOD) in the liVer tissue were also tested. Results ADP leVel was (7. 46±1. 12),(3. 58±0. 98),(4. 89±1. 02),(4. 79±1. 01) and (4. 13±0. 89) ng·mL-1 in N,M,NM,JH and JL grouPs, resPectiVely. The ADP leVel was significantly higher in grouP JH and JL than in grouP M (P<0. 01),and significantly higher in grouP JH than in grouP JL (P<0. 05). MDA leVel was (2. 98±0. 09),(4. 22±0. 11),(3. 66±0. 10),(3. 72±0. 11),(3. 99±0. 13) nmol·mL_1 in N,M,NM,JH and JL grouPs,resPectiVely. The MDA leVel was significantly lower in grouP JH and JL than in grouP M (P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). SOD leVel was (240. 8±17. 4), (149. 9±20. 6),(181. 6±19. 4),(209. 8±19. 2),(189. 4±18. 9) U·mL_1 in N,M,NM,JH,and JL grouPs,resPectiVely. SOD leVel was significantly higher in grouP JH and JL than in grouP M (P<0. 01),and significantly higher in grouP JH than in grouP JL (P<0. 05). TG leVel was (28. 98±1. 68),(214. 46±5. 44),(198. 46±6. 98),(142. 87±6. 64) and (164. 92±7. 56) mg·g-1 in N,M,NM,JH and JL grouPs,resPectiVely. TG leVel was significantly lower in grouP JH and JL than in grouP M ( P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). ALT and AST were significantly lower in grouP JH and JL than in grouP M (P<0. 01),and significantly lower in grouP JH than in grouP JL (P<0. 05). Conclusion The liPid Peroxidation in the liVer of rats with T2DM comPlicated with NAFLD can be reduced by gyPenoside,and hePatic lesion may be alleViated through inhibition of liPid Peroxidation.

Objective To get genetic information of VP 1 coding region of HEV 71 in Beijing in 2008.Methods Enteroviruses were isolated from samples of throat swabs collected from 33 HFMD patients within 3 days after onset by rhabdomyosarcoma(RD) cells and identified by RT-PCR method with specific primers to human enteroviruses,then VP 1 coding region was amplified and sequenced by Sanger dideoxy.Bioedit 7.0.5 and MEGA3.1 were used for the nucleotide sequence alignment and phylogenetic analysis.Results 16 virus strains were isolated from 33 samples,of which 14 strains were identified as HEV 71 and 2 were CVA 16,and 1 was co-infected with CVA 16 and HEV 71.Sequence analysis of VP 1 nucleotide sequences of 10 HEV 71 isolates showed that homologous analysis of nucleotide identity amino acid 95.5%～100% and 98.9%～100% respectively;representative strains of Fuyang in 2008,nucleotide identity was 95.4%～99.1%;with representative strain of C 4 nucleotide identity was over 92%.Phylogenetic analysis of HEV 71 strains for the nucleotide sequence of VP 1 coding region clarified that the HEV 71 isolates in Beijing belonged to C 4a cluster of C 4 sub-genotype and 10 strains formed four relatively separated clusters.Conclusions The HEV 71 viruses isolated from children of HFMD in Beijing belonged to C 4 sub-genotype,and C4a cluster which was the predominant in China since 2004.According to phylogenetic analysis,HEV 71 which belonged to more than 4 different clusters were circulating in Beijing in 2008.More virological suggestion for disease control and prevention,and information of HEV 71 molecular epidemiology need to be collected urgently due to the successive large epidemic of HEV 71 in China.

Adult ; Drainage ; Female ; Humans ; Hysterectomy, Vaginal ; adverse effects ; Laparoscopy ; adverse effects ; Radiography ; Subphrenic Abscess ; diagnostic imaging ; etiology ; therapy ; Ultrasonography

The clinical data of patients with hypoparathyroidism accompanied by elevated and normal muscle enzymes were analyzed retrospectively.The results showed that there were no differences in age,gender,pathogenesis,etiology,and parathormone level between groups with elevated muscle enzyme and normal muscle enzyme.Serum calcium concentration in the group with normal muscle enzyme was within normal range.Serum calcium concentration in the group with elevated muscle enzyme was significantly lower than that in the group with normal muscle enzyme (P＜0.01).As the serum calcium concentration rose,the elevated enzyme level gradually returned to normal range,suggesting that the decrease of serum calcium concentration resulted in the elevated muscle enzyme levels in patients with hypoparathyroidism.

Objective To construct the adenovirus vector containing mice CD40- IgG2aFc-IRS-GFP fusion gene and detect the expression of the fusion protein in transfeete 293cells.Methods The fragments of CD40, IgG2aFc were obtained and inserted into plasmid PDC316-IgG2aFc-GFP.After being verified by sequencing, Ad5-PDC316-CD40-IgG2aFc-GFP was contransfored with the bckbone vector into 293 cells.The virus titer was detected after replicating and purifing and the expression of the fusion protein was analyzed.Results A virus and plasmid were constructed successfully.The vitro infection showed that the virus can infect cells of which the fusion protein was confirmed by fluo-rescence.The expression of the fusion protein increased with the increased time and virus concentra-tion.The protein expression stopped increasing after the virus concentration came to a certain level.Conclusion CD40, IgG2aFc fragments are correctly ligated with plasmid PDC316-IgG2aFc-GFP, and the fusion protein can be expressed in 293cells.This might lay a foundation for further studies of the expresstion of virus, immune tolerance and mechanism of liver transplantation model in rats.

Objective To observe the effects of hepatitis B virus(HBV)X gene muhifunctional protein(HBx)on the biological characteristics of QSG7701 and the transformational effects on QSG7701 cell.Methods QSG7701 ceils were stably transformed by recombinant plasmid pCMV/X and eukaryotic expressed plasmid pRc/CMV2 by liposome-based assay,respectively.Non-transfeeted QSG7701 cells were employed as controls.The expressions of HBx,c-Myc and Bel-2 proteins in QSG7701 cells were detected by Western blot.MTT colorimetric analysis,flow cytometry and soft agar clone-forming assay were performed tO detect the biolo~dcal activity of cells.Results HBx Drotein was highly expressed in pCMV X/QSG7701 cells.The expression level of c-Myc protein in the pCMV X/QSG7701 cells was much higher than those in the other tWO groups of cells.The expression of Bcl2 protein was detected in the three groups of cells,but the expression levels were similar.Percentage of S stage cells in pCMV X/QSG7701 ceils was significantly higher than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells E(28.80±2.32)%,(15.5±2.64)%and(21.5±3.66)%,LSD 0.05=3.95%,LSD 0.01=5.47%,P<0.01].While percentage of G1 stage cells in pCMV X/QSG7701 cells was significantly lower than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells[(62.30±3.85)%,(78.70±4.12)%and(78.10q±4.45)%,LSD 0.05=5.63%,LSD 0.01-7.79 %,P<0.01].The apoptosis rate of pCMV X/QSG7701 cells was much higher than those in pRcCMV2/QSG770t and non-transfected QSG7701 cells[(14.90±1.01)%,(8.91±0.48)%and (4.03±0.47)%,LSD 0.05=O.94%,LSD 0.01=1.31%,P<0.01].The population doubling time of pCMV X/QSG7701 cells was shorter than those in pRcCMV2/QSG and non-trarmfected cells(14 h,29 h,38 h,respectively).The cloning ratio in soft agar of pCMV X/QSG7701 cells WaS significantly higher than those of pRcCMV2/QSG and non-transfected QSG7701 cells[(19.83±1.96)%,(1.76±0.03)%and (1.33±0.18)%,LSD 0.05=1.53%,LSD 0.01=2.11%,P<0.01].Conclusion HBx may transform human non-tumor hepatoeyte QSG7701,which makes the cell growth malignizeA.

Objective:To investigate the relationship between hypercoagulable state and the activation of the NF- kappa B pathway,inflammatory/suppression cytokines in patients with osteoarthritis. Methods:56 patients with OA were divided into two groups according to random number table:XFC group ( 28 cases ) and glucosamine ( GS ) group ( 28 cases ) . Two groups were treated for 3 months. Nine healthy people are healthy control group ( NC) . Determining the expression levels of the index of the NF-κB signaling pathway (p50,p65,TAK1,IκBα) and TNF-α,IL-1,IL-10,platelet activating factor(PAF) in serum by enzyme-linked immunosorbent assay. Detected the level of the indicators and laboratory indexes related with coagulation,observed the changes between the two group, used OA symptoms integral scale, LequesneMG, SF-36 and vas to assess efficacy;and made a correlation analysis. Results: After treatment,FIB,D-D, PAF, PLT, p50, p65, TAK1, IL-1, TNF-α, hs-CRP, ESR, IgG, LequesneMG, symptom integral meter, and VAS integral were significantly higher,APTT,PT,PAF-AH,IL-10 and each dimension integral of SF-36 significantly decreased in 2 groups (P<0. 05). XFC group was better than the GS group in reducing the level of PLT,FIB,TNF-α,p65,TAK1,hs-CRP,ESR,symptom integral meter,and VAS integral and increasing PT,each dimension integral of SF-36, etc (P<0. 05,P<0. 01). The results of Pearson correlation analysis show that PLT,FIB,D-D,PAF had positive correlation with p50,p65,TAK1,IL-1,TNF-α,hs-CRP,ESR,IgG,Le-quesneMG,symptom integral meter,and VAS integral,and negative correlation with IL-10 and each dimension integral of SF-36 ( P<0. 05,P<0. 01). PT had positive correlation with IL-10,GH and PF,and negative correlation with p50,p65,TAK1,IL-1,TNF-α,hs-CRP,ESR,IgG,symptom integral meter,and VAS integral (P<0. 05,P<0. 01). Conclusion: XFC could inhibit the NF-κB signaling pathway, raise the level of IL-10, reduce the expression of IL-1, TNF-α, P50, p65, TAK1 and so on, and reduce the abnormal inflammatory immune response. So as to achieve the purpose of delaying and inhibiting the production of hypercoagulable state,reduce joint disease,relieve the symptoms of joint pain and stiffness,eventually improve the patient’s quality of life.

Objective To investigate the effect of intermittent hypoxia on neuronal apoptosis and inducible nitric oxide synthase (iNOS) expression in rat hippocampus,in order to explore the potential mechanism of hippocampal neuronal apoptosis induced by intermittent hypoxia.Methods Twenty-four Wistar male rats (280-350 g) were randomly divided into three groups:the normal control,intermittent normoxia and intermittent hypoxia group (n=8 each).The animal model of intermittent hypoxia was established by an automated nitrogen/oxygen profile system.The three groups were respectively exposed to continuous normoxia (21 ％O2),cyclical normoxia (21 ％O2),and cyclical hypoxia [alternating between normoxia (21 ％ O2) and hypoxia (5 ％ O2),every 120 seconds] throughout the eight hours of light time(8:00-16:00).The rats were dissected and the hippocampus was removed at the end of the designated duration of exposures for six weeks.TdT-mediated dUTP nick end labeling (TUNEL) was used to detect the apoptotic rate of the hippocampus region in each group.Reverse transcription-PCR,immunohistochemistry (IHC) and Western blotting were used to examine mRNA and protein expressions of iNOS in the hippocampus tissue.Results The apoptotic rate of hippocampal neurons was higher in intermittent hypoxia group than in intermittent normoxia group [(28.236±0.081) ％ vs.(9.341±0.026)％,P＜0.05].The mRNA and protein expressions of iNOS were higher in intermittent hypoxia group than in intermittent normoxia group (3.394± 1.344 vs.0.125±0.040,7.793±0.052 vs.1.356±0.039,both P＜0.05].Conclusions Intermittent hypoxia can induce hippocampal neuronal apoptosis in rats,accompanied by the upregulation of iNOS gene transcription and protein expression,which indicates that iNOS may involve in the process of hippocampal neuronal apoptosis induced by intermittent hypoxia.