In this experiment, Hca-F or L615 elemene combo-TCV (H-TCV and L-TCV), H-TCV lysates, corynebacterium parvum (CP) were used to immunize 615 and Balb/c inbred mice, and their splenic DCs were prepared and pulsed in vitro with tumor cell lysates (H or L) and TCV lysates (TH or TL). The capacities of DCs to stimulate the proliferation of syngeneic nonadherent spleenic cells were tested with MTT assay. The results showed that the splenic DCs from normal mice in vitro pulsed with TH or TL could induced syngeneic noradherent splenic cells to proliferate (P＜0.01), while the H or L pulsed DCs could not. The splenic DCs from H-TCV or L-TCV or TH immunized mice re-pulsed in vitro with TH or TL exhibited stronger stimulating effects than the DCs from normal mice pulsed in vitro for the firth time pulsed with TH or TL (P＜0.01 or P＜0.05); The capacities of DCs to induce proliferation of syngeneic nonadherent splenic cells could be further enhanced by CP immunization, especially when were pulsed with TH (P＜0.01). Normal inbred 615 mice were transferred with DCs pulsed with lysates of elemene TCV (TDCs) or pulsed with lysates of Hca-F tumor cells (HDCs) on day 7 before challenged with lethal dose of live Hca-F cells, significant adoptive immunoprotective effects were seen, with 61.6% tumor inhibition rate and 25% survival in TDC adoptive transfer group. This study indicated that DCs might play a role in the mechanisms of active immunization and pulsing DCs with lysate of elemene combo TCV and isolating DCs from elemene combo TCV immunized mice were useful methods for DCs vaccine preparation.

Objective To explore the relationship between pulse pressure (PP) and artery atherosclerosis in elders. Methods Totally 2358 eiders( > 60y) with the risk factors of artery atherosclerosis from Yangpu District DOI:10.3760/cma.j.issn.1008-6315.2009.09.006Central Hospital and 11 Community Health Service Centers were enrolled . The basic information includes age, sex, height,body mass, history of smoking, blood pressure, heart rate, history of coronary heart disease, ischemic stoke, chronic kidney diseases and diabetes,etc. The fasting blood sugar(FBS) ,blood lipid( total cholesterel,triglyceride, low-density lipreprotein, high-deusity liproprotein), serum creatinine, serum uric acid, serum alanine aminotransferase were measured. The elders were divided into two groups according to the PP( PP <60 mm Hg and PP≥60 mm Hg) to analyse the relationship between PP and age, BMI, SBP, DBP, HR, FBS, TC, TG, LDL-C, HDL-C, Cr, Ccr, UA, AST and history of coronary heart disease,ischemic stoke,chronic renal disease, and diabetes. Results Serum levels of FBS,TC,TG, LDL-C, UA in the group with PP less than 60 nun Hg were ( 5.6±1.7 ) mmol/L, (4.3±1.3 ) mmol/L, ( 1.5±1.2) mmol/L, ( 2.5±1.1 ) mmol/L, UA ( 246.8±131.2 ) μmol/L, that in the group with PP over or equal to 60 mm Hg group were (6.3±2.6) mmol/L, (4.8±1.2) mmol/L, ( 1.9±1.1 ) mmol/L, (2.9±1.2) mmol/L, (291.4±133.6) μmol/L, Serum levels of FBS, TC, TG, LDL-C, UA in the group with PP over 60 mm Hg were obviously elevated as compared with that in the group with PP lower than 60 ram Hg(P <0.01 ) ; The preven-lence rates of coronary heart disease ischemic stoke,chronic,renal disease, diabetes in the group with PP less than 60 mm Hg were 10.8% ,26.0%, 12.4%, 17.6%, that in the group with PP over or equal to 60 mm Hg group were 17.8% 、31.7% ,16.9% ,23.8% (P <0.01 ). The preveulence of coronary heart disease ischemic stoke,chronic re-nal disease, diabetes were significandy increased in the group with PP over or equal to 60 mm Hg group. Logistic re-gression analysis showed PP was closely related to FBS, BMI, age, LDL-C, UA, HDL-C ( β = 0.103,0.093.0.097, 0.089 ,0.076 , - 0.057, P < 0.05 ). Conclusions For those elders with the basis of artery atherosclerosis, high PP can inhance the development of artery atheresclerosis and also inhance the occurance and development of cardio-ce-rebrovascular and renal diseases.

Objective To explore the operative methods and clinical outcomes in emergency or sube- mergency repair for the complicated tissue defects in hand in first stage applying microsurgical technique. Methods From Jan.,2000 to Aug.,2005,49 emergency cases of complicated hand tissue defects were re- paired in the first stage with replantation,reconstruction,free flaps,combined finger reconstruction and flap transplantation,including 21 cases in mini tissue mass replantation or reconstruction,15 cases in replantation combined with free flap transplantation,8 cases in replantation and reconstruction combined with free flap transplantation,5 cases in combined multiple digits reconstruction with free flap transplantation.The free flap transplantation included the anterolateral femoral flap,the latissimus dorsi myocutaneous flap,the dorsalis pe- dis flap,the media pedis flap and the instep flap.Results All the flaps,the replanted and reconstruceted finger survived uneventfully except for one replanted finger necrosis.45 cases healed in the first stage and the other 4 cases healed in the second stage.During a follow-up from 6 months to 3 years postoperatively,a satis- factory appearance and function of the reconstructed hand was achieved.The excellent and good rate was 85.7% assessed with provisional functional assessment criterion for upper limbs issued by Chinese Society of Hand Surgery.Conclusion The emergency or subemergency repair for the complicated tissue defects in hand has the advantage of short-term treatment and desirable functional outcome.The emergency replantation and reconstruction combined with various flaps or tissue mass can be applied to repair tissue defect in hand in the first stage according to the position and area of the defect along with the technique level of the surgeon, having been proved to achieve desirable clinical outcomes.And the key points leading to a successful operation is the correct treatment for the raw surface of the defects,suitable choice for various flaps,logical design of combination pattern and prevention and timely treatment for vessel crisis.

ObjectiveTo evaluate the potential value of acoustic radiation force impulse (ARFI)elastography in the characterization of solid liver tumors.MethodsForty-three patients with 56 liver tumors were evaluated with ARFI,which included 21 patients with hepatocellular carcinoma (HCC),8 patients with metastase,5 patients with cholangiocarcinoma(CCC),and 9 patients with hemangioma.The shear wave velocity of the tumor and background liver parenchyma were calculated,and results were compared with 30 healthy subjects.Statistical analysis was performed on the shear wave velocity for differentiation of normal liver,background liver parenchyma,and tumors.ResultsHCC and CCC had greater stiffness than metastase (P ＜0.05),there were no statistical differences between HCC and CCC (P = 0.179).Malignant liver tumors had significantly greater stiffness than hemangioma and normal liver (P = 0.000).34.5% (9/26) HCC and 33.3% (4/12) hemangioma appeared softer than the background liver.With a cut-off value of 1.5 m/s for the shear wave velocity,the sensitivity,specificity,positive predictive value and negative predictive value for malignancies were 79.5%,83.3%,94.5% and 52.6%,respectively.ConclusionsARFI elastography quantification is a promising noninvasive technique for assessing solid liver tumors.Use of ARFI elastography quantification may lead to new quantitative tissue characterization parameters for differentiating hemangioma and malignant liver tumors.

OBJECTIVETo evaluate the feasibility and safety of laparoscopic and endoscopic cooperative surgery for treating gastric gastrointestinal stromal tumors(GIST).

METHODSRetrospective analysis was performed on the clinical data of 46 patients with gastric GIST undergoing laparoscopic and endoscopic cooperative surgery between June 2009 and June 2011 at the Renji Hospital of Shanghai Jiaotong University School of Medicine.

RESULTSThere were 27 males and 19 females with the mean age of 58.5 years. Thirty-three patients received endoscopy-assisted wedge resection, and 13 cases received laparoscopy-assisted endoscopic resection. All the operations were successful. The mean operative time was (85.5±29.3) min, the mean blood loss was (31.4±12.2) ml, the mean post-operative gastrointestinal functional recovery time was (31.6±14.9) h, and the mean post-operative hospital stay was (5.1±2.9) d. No post-operative complication occurred. NIH risk assessment showed that 34 cases were very low risk and 12 low risk. No recurrence or metastasis was found during the follow-up ranging from 2 to 26 months(median, 12.6 months).

CONCLUSIONLaparoscopic and endoscopic cooperative surgery for gastric GIST is both feasible and safe with minimal invasiveness, fast recovery and satisfactory short-term outcomes.

Adult ; Aged ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; surgery ; Gastroscopy ; Humans ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Treatment Outcome

BACKGROUNDPrevious studies have suggested that interrupted clearance of nuclear DNA-protein complexes after cell death might initiate and propagate systemic lupus erythematosus (SLE). Deoxyribonuclease I (DNaseI) may be responsible for the removal of DNA from nuclear antigens at sites of high cell turnover, thus preventing the onset of SLE. The purpose of this study was to genotype the single nucleotide polymorphisms (SNPs) of DNase1 and characterize its gene expression and alternatively spliced transcripts in Chinese patients with SLE in order to understand the pathogenic role of DNase1 in human SLE.

METHODSFour SNPs located at the 3' end of the DNase1 gene, as listed on the SNP website, were selected for analysis. Those SNPs with relatively high heterozygosity were chosen for genotyping in 312 Chinese SLE families using the Taqman minor groove binder (MGB) allelic discrimination method. Haplotypes were constructed and linkage disequilibrium tests were performed using GeneHunter. DNase1 mRNA expression was detected using real-time polymerase chain reaction (PCR), and alternatively spliced transcripts were isolated using capillary electrophoresis. Any effects the specific SNP haplotypes had on DNase1 gene expression and the alternatively spliced transcripts were also assessed.

RESULTSrs179982 and rs1053874 had high heterozygosity, about 0.5 in this Chinese cohort, while rs1059857 was also found to be heterozygous. Analysis of the haplotype combining rs179982-rs1030874 (C-G) and rs179982-rs1030874-rs1059857 (C-G-G) revealed a skewed transmission in favor of affected offspring. DNase1 gene expression was higher in SLE patients than in normal controls (P < 0.001), but this was not related to disease activity or SNP haplotype. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients differed from that of normal controls. Furthermore, different SNP haplotype combinations generated different transcript patterns in SLE patients.

CONCLUSIONSThe SNP haplotypes are in linkage disequilibrium in Chinese SLE patients and may induce the disease through a modification of DNase1 mRNA splicing rather than at the level of mRNA expression. There is a relatively unique transcript band in SLE patients independent of special haplotype, which suggests that other unknown factors might be involved in adjusting gene expression.

Adolescent ; Adult ; Alternative Splicing ; Deoxyribonuclease I ; genetics ; Female ; Haplotypes ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

OBJECTIVETo localize susceptibility loci in Chinese systemic lupus erythematosus (SLE) cohort by linkage disequilibrium mapping the genomic interval in human chromosome 16 so as to understand whether the pathogenesis of human SLE is related to chromosome 16.

METHODSFive microsatellite markers in chromosome 16 spanning from 57.79 cM to 65.1 cM were used for fluorescence based genotyping in 157 SLE families. Evidence for linkage disequilibrium was assessed using the extended transmission disequilibrium test (ETDT) program and Genehunter software. Susceptibility gene was searched in the positive locus, and real-time PCR method was used to detect gene expression.

RESULTSWith the ETDT, evidence for linkage disequilibrium of the marker D16S409 (P=0.0278) and D16S517 (P<0.0001) with SLE were found. It was observed that 271 bp allele of D16S517 was much in evidence for preferential transmission, while 277 bp allele was found to be transmitted less often than expected. Genehunter analysis is concordant with ETDT. By real-time PCR, OAZ(OLF1/ EBF-associated zinc finger protein) gene which is located in the positive locus showed higher expression in SLE patients than in normal controls.

CONCLUSIONResults reveal that 16q12 (58.46 cM) is associated with Chinese SLE. OAZ is a likely susceptibility gene in this locus for SLE.

Adolescent ; Adult ; Alleles ; China ; Chromosomes, Human, Pair 16 ; genetics ; Cohort Studies ; DNA ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Microsatellite Repeats ; Middle Aged

OBJECTIVETo observe whether deoxyribonuclease I (DNASE1) gene expression and its DNASE1 mRNA expression was detected by real-time polymerase chain reaction and its alternatively spliced transcripts were performed by capillary electrophoresis. An analysis was also made to disclose whether specific single nucleotide polymorphisms (SNPs) haplotype had effects onDNASE1 gene expression and its alternatively spliced transcripts.

RESULTSDNASE1 gene expression was higher in SLE patients than in normal controls (P<0.001), and in patients it was found to be of no relationship with SLE disease activity index score. However, it was increased in female patients. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients was not the same as that in normal controls. Moreover, it seemed that different SNPs haplotype combination might show different transcript pattern in SLE patients.

CONCLUSIONIn SLE patients, DNASE1 gene expression is abnormal and there are alternatively spliced transcripts different from those in normal controls. DNASE1 gene is a critical factor in the pathogenesis of SLE.

Adolescent ; Adult ; Alternative Splicing ; Deoxyribonuclease I ; genetics ; Female ; Gene Expression ; Humans ; Lupus Erythematosus, Systemic ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide

OBJECTIVETo study the expression level of cyclin D1-CDK4 protein in human embryonic lung fibroblasts (HELF) induced by quartz, and to study whether the expression level of cyclin D1-CDK4 protein mediated by mitogen activated protein kinase (MAPK)/(AP-1) signaling pathways.

METHODSCells were harvested after stimulation 2 h for the detection of cytokines. Cyclin D1 and CDK4 (cyclin dependent kinase 4) proteins was measured by immunocytochemistry (IC) and Western blot (WB).

RESULTSThe exposure of HELF to crystalline quartz for 2 hours could cause the decrease of cyclin D1 and cyclin dependent kinase 4 (CDK4) protein expression level, (7.91 +/- 0.29) x 10(3) and (5.17 +/- 0.28) x 10(4) respectively, which was lower than that of the HELF group (P < 0.05). AG126 (chemical inhibitor of the extracellular signal regulated protein kinase (ERK) signaling pathway) and the dominant negative mutant of ERK2 (molecular inhibitor of ERK2), prevented the decrease of cyclin D1 and CDK4 protein expression level. The chemical inhibitor of c-Jun NH2-terminal amino kinase (JNK), SP600125, could prevent both cyclin D1 and CDK4 protein expression level decrease. But SB203580, the chemical inhibitor of p38, prevented neither cyclin D1 nor CDK4 protein expression level decrease. Curcumin could prevent CDK4 protein expression level decrease but not cyclin D1 protein.

CONCLUSIONERKs and JNKs, but not p38, are responsible for the decrease of cyclin D1 and CDK4 protein expression level in HELF induced by quartz. AP-1 is responsible for the decrease of CDK4 expression level but not that of cyclin D1.

Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Humans ; Lung ; cytology ; embryology ; Mitogen-Activated Protein Kinases ; metabolism ; Quartz ; toxicity ; Transcription Factor AP-1 ; metabolism

Objective To study the routine examination of Aconitum sinomontanum Nakai and determine the contents of lappacontine and ranaconitine; To provide basis for establishing the quality standard of Aconitum sinomontanum Nakai.Methods Aconitum sinomontanum Nakai were collected from different areas.A method of TLC was used for qualitative discrimination. The methods in the Chinese Pharmacopoeia were adopted for the determination of moisture content, ash content and extractives. Determination of lappacontine and ranaconitine were performed by HPLC. Results The TLC showed that the spots were clear and the separation was good. Individual provisional standards:the moisture,total ash and acid-insoluble ash content of Aconitum sinomontanum Nakai were not more than 11.0%, 12.0%, and 7.0%, respectively; water soluble and alcohol soluble extractives were not less than 18.2% and 10.6%,respectively.The content of ranaconitine and lappacontine in Aconitum sinomontanum Nakai were not less than 0.125% and 0.815%, respectively. Conclusion The method established by the study is accurate and reliable,and can be used for quality evaluation of Aconitum sinomontanum Nakai.