Objective:To investigate the strategies and methods of pancreatic cancer immunotherapy adopt cytotoxic T lymphocytes activated by dendritic cells(DCs),which modified by heat shock protein 70 peptide complex(HSP70-PC).Methods:HSP70 peptide complex from lumps of tumor-bearing mouse innoculated with pancreas cancer cell(MPC83)were purified by ConA-Sepharose and ADP-Agarose affinity chromatography.Dendritic cells derived from normal murine bone marrow were induced by GM-CSF and IL-4.MTT method was applied to analyses the proliferation activities of DCs.To achieve the specific T lymphocyte cells,DCs modified with HSP70 peptide complex were coculture with murine spleen lymphocytes for 48 hours.Then these activated lymphocytes were cocultured with MPC83 at ratio of effect-target 10∶1 and 20∶1 in vitro and the killing activities were detected by MTT method.Results:High purify HSP70-PC was obtained;104 DCs need 50～100ng of HSP70-PC to modify,100?g of HSP70-PC peptide complex can be extracted from per 1g lump;T cells stimulated by HSP70-PC modify DCs showed more specific cytotoxic activity to MPC83 than lymphocytes activated by DCs without HSP70-PC in vitro.Conclusion:We can extract high purity HSP70-PC for clinical individual immunotherapy from pancreatic cancer lump by using hypobaric affinity chromatography;DCs vaccine modified with HSP70-PC has significant kill effect against pancreatic cancer cell in vitro.

Objective To discuss the change of cellular immune and humoral immune function in children patients with Henoch-Schonlein purpura(HSP)and to investigate its clinical significance.Methods The expression levels of T lymphocyte CD3 + ,CD3 +CD4 + ,CD3 + CD8 + ,CD3 + CD4 + CD8 + ,B lymphocyte CD3 - CD19 + and NK lymphocyte CD3 - CD16 + CD56 + in periphetal blood of 156 patients with HSP was detected by flow cytometry.The level of immunioglobins(IgG,IgA,IgM)and complement(C3,C4)of serum were evaluated by nephelometry immunoassay.The level of immunioglobin IgE was measured by immunochemiluminometric assays and compared with 25 healthy controls.Results Compared with control group,the ratio of CD4 +/CD8 + and the contents of CD3 + ,CD4 + cells in HSP patients were decreased significantly(P <0.01),while the content of CD19 + was significantly increased (P <0.01).The content of CD16 + CD56 + was decreased(P <0.05 ),while the content of CD3 + CD4 + CD8 + was increased(P <0.05),the content of CD8 + had no obvious change(P >0.01).The level of IgA and IgE were increased significantly(P <0.01).The level of IgG and C3 were increased (P <0.05),The level of IgM and C4 had no obvious change(P >0.05).Conclusion The cellular immune and humoral immune function in children with HSP are in disorder.To detect lymphocyte subsets,immunioglobin and com-plement will be of important significance to understand the immune function from the perspective of the cellular immune and humor-al immune to explore the pathogenic mechanisms and to assist diagnosis and patient′s prognosis.

Objective To investigate the clinical and laboratory features of acute promyelocytic leukemia (APL).Methotis 513 APL patients in the last two decades were retrospectively analyzed in this research.We investigated the clinical features including age,sex,abnormality of peripheral hemogram before treatment.therapeutic effect and follow-up and laboratory data such as morphology,immunology,cytogenetics and molecular biology(MICM).Results The median age of the APL patients was 33 years old and the ratio of male and female was 1.21:1.Before treatment,the median level of WBC was 4.3×109/L and the deteetion rate of abnormal promyelocyte on blood film was 85.8%;with immunophenotypie detection,the expression levels of CD117、CD34、HLA-DR、CD7、CD14 and CD19 in APL were found to be lower and the expression 1evels of CD2、CD33 and MPO higher than those in other subtypes of acute myelocytie leukemia(AML)(beth P<0.01).Specific abnormal chromosome t(15;17)was detected in 91.7%of the patients,of whom 75.9%had standard translocation of t(15;17),being the most common one and 15.8% of the patients had t(15;17)with additional abnormal chromosome.There was only 7.5%of the patients with nolnlal karyotype.However,the presence of both simple translocation and complex translocation was seldom seen.With molecular biological detection.PML/RARα fusion gene positive rate was 99.6%.In a relativelv long clinical follow-up,we found that the complete remission(CR)rate in APL patients was 84.7%.incidence of DIC was 13.4%and five-year survival rate was 30.7%.111e median count of WBC in CR group was lower than that non-remission group(P<0.01).There were no significant differences on expressions of CD34 and CD2 and changes of cytogenetics between the two groups(P>0.05).Conclusions Comprehensive evaluation of MICM could be of important significance in the diagnosis and prognosis iudgrnent for APL patients.The CR rate in these patients with high WBC eount was considerable low.

Objective To study the changes and significance of CD4+CD25+ regulatory T cells and C-reactive protein(CRP) in patients with acute exacerbation of chronic obstructive pulmonary disease(COPD). Methods Flow cytometry was used to detect the frequency of CD4+CD25+ regulatory T cells in peripheral blood from 36 patients with acute exacerbation of COPD( COPD group) and 36 patients with clinical stability of COPD(control group one)and 36 normal individuals(control group two). The level of CRP was detected routinely. Results The ratio of CD4+CD25+ regulatory T cells number in peripheral blood of COPD group to the total number of CD4+T cell was (2.56±1.83 )%, and it was significantly decreased compared to the other two groups (P all<0.01 ). The level of CRP in COPD group was markedly higher than that in the other two groups (P all<0.01 ). The level of CD4+CD25+ regulatory T cells in patients with acute exacerbation of COPD had negative relation with CRP. Conclusions CD4+CD25+ regulatory T cells participate in inflammatory response. The proportion of CD4+CD25+ regulatory T cells decreases in patients with acute exacerbation of COPD, and it may result in maladjustment of cytoimmunity.

Objective To evaluate the immune reconstitution of HIV-1 infected individuals after long-term highly antiretroviral therapy (HAART). Methods Twenty-five HIV-1 infected individuals receiving HAART, 17 without HAART and 15 healthy controls were included in the study. CD4 +T, CD8 +T,CD8/human leukocyte antigen DR (HLADR) + T, CD8/CD38 +T cells, and the expression of CD127 on CD3 +T cells from peripheral blood samples were measured by flow cytometry. IL-7 in peripheral blood was measured by enzyme-linked immunosorbort assay (ELISA) in HAART group. t test was performed to compare the measurement data among the groups. Results Before HAART, the count of CD4 + T cells in HIV-1 infected group was lower than that of the healthy control (t =9. 12, P ＜0. 01 ), while the counts of CD8 + T,CD8/HLADR+T, and CD8/CD38 +T cells were higher than those of the healthy control (t = 4.48, 4.89 and 3.88, P＜0. 01 ). Ater 7 years' antiviral therapy, CD4 +T cells increased, CD8 +T cells decreased, but both of them didn' t reach to the normal levels ( t = 2.66 and 2.43, P ＜ 0.05 ). While the counts of CD8/HLADR+T cells and CD8/CD38 +T cells almost reached to the normal levels (t = 0. 86 and 1.39, P ＞0.05). Before HAART, the concentration of IL-7 in HIV-1 infected group was higher than that in healthy controls (t =5.31, P ＜0.01 ). It decreased with HAART, but was still higher than the normal level (t =2. 81, P ＜ 0. 05 ). The expression of CD127 on CD3 + CD8 + T cells in non-HAART group was significantly lower than that in healthy control ( t =- 6.01, P ＜ 0.01 ), while that in HAART group was higher ( t = 2.32,P ＜0.05), but still not reached to the normal level ( t = 4.49, P ＜ 0. 05 ). CD127 expression on ( CD45RA + ) CD3 + CD8 + T cells almost increased to the normal level ( t= 0. 28, P ＞ 0. 05 ), while that on ( CD45RO + ) CD3 + CD8 + T cells was still remarkably lower than the normal ( t = 4. 86, P ＜ 0. 05 ). Conclusion Long-term HAART can partially restore the count and function of lymphocyte subsets in HIV-1 infected individuals, and the abnormal immune activation can be inhibited.

[Objective] To establish the quality standard for Guanxin Qiwei Tablet. [Methods] Radix Salviae Miltiorrhizae, Lignum Dalbergiae Odoriferae, Rhizoma Kaempferiae and Fructus Choerospondiatis in Guanxin Qiwei Tablet were identified by TLC. TanshinoneⅡA content was determined by HPLC.[Results] Radix Salviae Miltiorrhizae, Lignum Dalbergiae Odoriferae, Rhizoma Kaempferiae and Fructus Choerospondiatis can be identified by TLC, the spot being clear without the interference of negative control. A good linearity was in the range of 0.022-0.154 ?g, the average recovery of tanshinone ⅡA was 97.9%, and RSD was 1.22%. [Conclusion] This method is simple and can be used to evaluate the quality of Guanxin Qiwei Tablet.

Objective The aim of this study is to analyze the long-time outcome of hepatocellular carcinoma(HCC)patients with micro-vascular invasion underwent liver resection combined with transarterial chemoembolization(TACE).Methods Our database of surgical re-section from January 2009 to September 2015 was retrospectively analyzed.This study was conducted on 296 HCC patients with MVI.Patients were divided into two groups:one group underwent liver resection (n =159)and another for liver resection combined with TACE (n =137). The 5-year overall survival rate (OS)and disease free survival (DFR)were compared.A multivariate Cox proportional hazards regression a-nalysis was performed to assess the prognostic risk factors associated with overall survival rate.Results The 5-year OS and 5-year DFR see significant difference (OS:18% vs.8%,P =0.001;TRF:15% vs.8%,P =0.008).Multivariate analysis revealed that HBsAg(HR 1.596, P =0.002,95% CI 1.194 ~2.131),tumor size >5 cm(HR 0.729,P =0.042,95% CI 0.539 ~0.989)as well as multiple tumors(HR 1.480,P =0.049,95% CI 1.002 ~2.186)were correlated to poor overall survival rate.Conclusion Surgical resection combined TACE for HCC patients with MVI realized a better prognosis than patients merely underwent therapy of resection.

Objective To explore the indications for liver transplantation among patients with hepatolithiasis.Methods Data from 1 431 consecutive patients with hepatolithiasis who underwent surgical treatment from January 2000 to December 2006 were retrospectively collected for analysis.Surgical procedures included T-tube insertion combined with intraoperative cholangioscopic removal of intrahepatic stones,hepatectomy,cholangiojejunostomy,and liver transplantation.Results Nine hundred and sixty-one patients who had a stone located in the left or right intrahepatic duct underwent hepatectomy or T-tube insertion combined with intraoperative cholangioscopic removal of intrahepatic stones.The rate of residual stones was 7.5%(72/961).Four hundred and seventy patients who had a stone located in the bilateral intrahepatic ducts underwent surgical procedures other than liver transplantation;the rate of residual stones was 21.7%(102/470).Only 15 patients with hepatolithiasis underwent liver transplantation;they all survived.According to the degree of biliary cirrhosis,recipients were divided into 2 groups: a group with biliary decompensated cirrhosis(n=7),or group with biliary compensated cirrhosis or noncirrhosis group(n=8).There were significant differences in operative times,transfusion volumes and blood losses between 2 groups(P

OBJECTIVETo investigate the role of retinoic acid receptor beta (RARbeta) in mediating inhibitory effect of all-trans retinoic acid (ATRA) on activator protein-1 (AP-1) activity in gastric cancer cells.

METHODSTransient transfection and chloramphenicol acetyltransferase (CAT) assay, Nort hern blot, gene transfection, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, and anchorage independent growth assay were used.

RESULTSTransient transfection of RARbeta expression vector into MKN-45 cells resulted in the RARbeta concentration dependent repression of AP-1 activity induced by 12-o-tetradecanoylphorbol-13-acetate (TPA), regardless of the presence of ATRA. When the c-jun and c-fos expression vectors were cotransfected with the RARbeta expression vector into MKN-45 cells, AP-1 activity was also obviously repressed. The inhibitory effect, again, was RARbeta-concentration-dependent. The stable transfection of the RARbeta gene into MKN-45 cells led to cell growth inhibition and colony formation inhibition by ATRA. Furthermore, Cotransfection of both RARbeta/DNA binding domain (DBD) and reporter gene could not alter AP-1 activity, even in the presence of ATRA.However, when the cotransfection was substituted with the RARbeta/ligand binding domain (LBD), the inhibition was significantly enhanced by ATRA.

CONCLUSIONRARbeta might be required for anti-AP-1 activity, and contribute to growth inhibition of gastric cancer cells by ATRA.

Antineoplastic Agents ; pharmacology ; Binding Sites ; Cell Division ; drug effects ; DNA ; metabolism ; Humans ; Receptors, Retinoic Acid ; chemistry ; physiology ; Stomach Neoplasms ; drug therapy ; pathology ; Transcription Factor AP-1 ; antagonists & inhibitors ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

Objective To evaluate the changes in motor function impairment and brain functional networks of patients with acute ischemic stroke(AIS)through parameters of spontaneous activities of both upper limbs and electroencephalogram graph theory analysis methods.Methods The data of 34 acute ischemic stroke patients(observation group)with upper limb motor disorders who were treated in the Department of Neurology of the Affiliated Hospital of Southwest Medical University from January 2022 to October 2023,and 40 healthy controls(HC group)were collected.The subjects completed the National Institutes of Health Stroke Scale(NIHSS)and Fugl-Meyer Assessment(FMA)within 7 days,and wore wrist activity recorders(Actiwatch)continuously for 24 hours to collect data on sponta-neous activities of upper limbs and analyzed related parameters such as the coordination coefficient of both upper limbs(r),the activity ratio of the affected side to the healthy side upper limb(ULAR),etc.At the same time,all subjects completed approximately 2 hours of 19-channel electroencephalogram examination.After preprocessing the electroencephalogram data,5 segments of 10-second resting-state electroencephalogram were extracted for graph theory.Results ① Compared to healthy individuals,AIS patients exhibited decreased functional connectivity edges in the 8 and θ bands,with substantial reductions in network connections in the α band.In the β band,connections between the frontal,right parietal,and occipital regions weakened,while connections from the right temporal lobe to the left temporal lobe strengthened.In the γ band,there was a significant increase in connections throughout the brain.② Graph theory analysis revealed significantly increased shortest path lengths(α band:t=2.228,P＜0.05,d=-0.52;β band:t=-3.641,P＜0.01,d=-0.878)and decreased global efficiency(α band:t=2.535,P＜0.05,d=0.591;β band:t=3.321,P＜0.01,d=0.803)in the observation group compared to the control group.In the γ band,local effi-ciency(t=3.279,P＜0.01,d=0.765)and clustering coefficients were significantly higher(t=3.358,P＜0.01,d=0.783).③ In the-γ band,the ULAR ≤30％group showed significantly re-duced shortest path length(t=-2.063,P＜0.05,d=-0.802)and increased global efficiency(t=2.226,P＜0.05,d=0.865),local efficiency(t=2.95,P＜0.05,d=1.147),and cluste-ring coefficient(t=2.962,P＜0.05,d=1.148).④ In the observation group,the bilateral upper limb coordination coefficient during sleep was negatively correlated with NIHSS scores(r=-0.389,P＜0.05)and ULAR(r=-0.395,P＜0.05),while FMA scores were positively cor-related with ULAR(r=0.442,P＜0.05).Conclusion The parameters of spontaneous activities of the upper limbs can be used to determine the impairment of motor function in AIS patients.The combination of changes in brain functional networks and motor impairments can provide new ideas for the study of their neural network mechanisms.