Objective To investigate the survival of islet isograft in NOD mice treated with islet antigen-specific regulatory T cells.Methods GAD-65 antigen pulsed immature dendritic cells (imDC) were used to induce naive T cells into islet antigen-specific regulatory T cells.NOD mice which had progressed to type 1 diabetes (T1DM),as the recipients,received islet isografts (500 IEQ) under renal capsule from NOD mice without T1DM.In NOD mice in control group without transplantation,the changes in blood glucose (BG) were observed.NOD mice in simple islet transplantation group were given islet isograft without Treg infusion.In experiment group,NOD mice were infused with 1 × 106 islet antigen-specific regulatory T cells on the 1st day before transplantation,subsequently underwent islet isotransplantation.The survival of the islet isograft was evaluated by BG levels and the pathological changes were observed.Results BG levels were sustained above 11.1 mmol/L in control group.In simple islet transplantation group,BG level was decreased to the normal level in 1 ～2 days after transplantation,and began to rebound in 7～ 17 days posttransplantation and maintained at the preoperative level.The mean survival of the islet isograft in the NOD mice was (12.2 ± 2.6) day;In experiment group,BG level was decreased to the normal level in 1 ～2 days after transplantation,rebounded above 11.1 mmol/L in some mice on the 27th day after transplantation,and rebounded above 11.1 rnmol/L on the 43th day in all mice.The mean survival of the islet isograft in the NOD mice was (35.2 ± 4.3) days,which was significantly prolonged compared to simple islet transplantation group (P＜ 0.01).In simple islet transplantation group,the islet isograft was infiltrated by many lymph cells and damaged severely,and only few residual islet cells secreted insulin without complete islet existing in insulin staining.The islet isograft in experiment group was intact on the 15th day,with little lymph cell infiltration and a great number of islets secreting insulin.Conclusion Infusion of islet antigen-specific regulatory T cells induced by imDC and islet antigen GAD-65 in vitro could delay the destruction of autoimmune system and prolong the islet isograft survival in NOD mice.

Objective To develop a candidate reference method for the determination of sodium in serum by inductively coupled plasma mass spectrometry method (ICP-MS). Methods Aluminum, as internal standard of sodium, was added into serum samples and sodium standard solutions by gravimetric analysis. The samples were digested by HNO3 and diluted, and its 23Na/27Al isotope ratios were obtained by ICP-MS. The sodium concentrations were calculated with the standard curve method in serum. Results The analytical recoveries of sodium were 100.67% and 100.15% respectively, and the precisions were 0.08% and 0.04% respectively for 2 different serum samples. The results of measuring sodium in serum of Standard Reference Material (SRM) gave the coefficients of variation (CVs) of 0.18% and 0.22% for 2 levels of standard reference material(SRM) 909b and 0.41%, 0.41% and 0.66% for 3 levels of SRM 956b. The relative deviations between the results and median of the certified value were 0.17% and 0.14% for 909b and -0.09%, -1.05%, -0.48% for 956b respectively. Conclusions The ICP-MS and aluminum internal standard method is proved to be not only precise and accurate, but also quick and convenient for measuring sodium in serum. It is promising to be a candidate reference method for determination of sodium in serum.

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.

Objective:To investigate the effects of oxymatrine(OM)on the apoptosis of human hepatoma HepG2 cells and its possible mechanisms.Methods:HepG2 cells were treated with different concentrations of OM.The proliferation inhibition was measured by MTT assay and the apoptosis of HepG2 cells were examined by Hochest staining method.Flow cytometry was used to analyze the cell cycle distribution and apoptosis rate.The expression of caspase-3,Bcl-2,Bcl-x_L and Bax proteins was assayed by Western blotting assay.Results:OM inhibited HepG2 cells growth in a time-and dose-dependent manner.After treatment with OM for 24 hours,some cells appeared typical apoptotic characteristics and the apoptosis rate was increased. Treatment with OM also increased caspase-3 activity and Bax expression in HepG2 cells,and decreased the expression levels of Bcl-2 and Bcl-x_L.Conclusion:OM can induce HepG2 cell apoptosis,which may be related to the down-regulation of PI3K/Akt signal pathway,suppression of Bcl-2 and Bcl-x_L activity,and activation of caspase-3.

A phytochemical investigation on the roots and stems of Litsea cubeba led to the isolation of seven isoquinolone alkaloids. By spectroscopic analysis and comparison of their 1H and 13C-NMR data with those in literatures, these alkaloids were identified as (+)-norboldine (1), (+)-boldine (2), (+)-reticuline (3), (+)-laurotetanine (4), (+)-isoboldine (5), (+)-N-methyl-laurotetanine (6), and berberine (7), respectively. Among them, 7 was isolated from the genus for the first time. The evaluation of these compounds showed weak anti-inflammatory activity against NO production in RAW 267.4 and BV-2 cells.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Litsea ; chemistry ; Molecular Structure ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo study the expression of the Trp-p8 protein in the prostate tissue of the PSA "grey zone" with different PSA density of the transition zone (PSADTZ) and explore the value of determining Trp-p8 expression and PSADTZ in the early diagnosis of prostate cancer (PCa).

METHODSThis study involved 30 cases of benign prostatic hyperplasia (BPH) and another 30 cases of PCa with different PSADTZ values. Using a data imaging and analysis system, we determined the expression levels of Trp-p8 in BPH and PCa tissues and analyzed their correlation with PSADTZ.

RESULTSThe expression of Trp-p8 was weak or negative in the BPH but strong in the PCa tissue and even stronger in the PCa tissue with high PSADTZ (F = 34. 05, P < 0.05).

CONCLUSIONThe Trp-p8 protein is expressed differently in BPH and PCa tissues of the PSA " grey zone" and its expression is positively correlated with PSADTZ. Determination of the Trp-p8 expression and PSADTZ contributes to the early diagnosis of prostate cancer.

Biomarkers, Tumor ; metabolism ; Early Detection of Cancer ; methods ; Humans ; Male ; Prostate ; metabolism ; Prostate-Specific Antigen ; metabolism ; Prostatic Hyperplasia ; diagnosis ; metabolism ; Prostatic Neoplasms ; diagnosis ; metabolism ; TRPM Cation Channels ; metabolism

Adenoma ; genetics ; metabolism ; pathology ; surgery ; Adult ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Diagnosis, Differential ; Exons ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Mutation ; Nuclear Proteins ; metabolism ; Proto-Oncogene Proteins B-raf ; genetics ; Thyroglobulin ; metabolism ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Thyroid Nuclear Factor 1 ; Transcription Factors ; metabolism

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.

OBJECTIVETo explore the possible mechanism of electroacupuncture preconditioning (EAPC) and combined with ATP-sensitive potassium channel (KATP) blocker preconditioning for hypoxia/ischemic brain injury protection by observing the changes of the immediate genes (c-fos and c-jun protein content) in brain at the early stage after cerebral hypoxia/ischemic injury, and the effect of EAPC on these changes.

METHODSIntegrated density (ID) of c-fos and c-jun expression was measured by Western blot and computerized image processing.

RESULTSHypoxia/ischemia could induce c-fos and c-jun protein in both cerebral cortex and hippocampus simultaneously, with the peak appearing 2-4 hrs later, and the expression in hyppocampus was higher than that in cortex. EAPC could lower KATP blocker induced permanent high expression in hyppocampus.

CONCLUSIONThe effect of EAPC preconditioning in antagonizing cerebral hypoxia/ischemic injury may be related with its action in activating KATP, inhibiting the neuron apoptosis induced by the immediate genes at early stage of injury.

Animals ; Animals, Newborn ; Apoptosis ; Brain ; metabolism ; pathology ; Electroacupuncture ; Female ; Hypoxia-Ischemia, Brain ; metabolism ; prevention & control ; Ischemic Preconditioning ; methods ; Male ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Litsea cubeba is one of aromatic medicinal plant belonging to family Lauraceae. The roots, stems and fruits of L. cubeba have been widely applied as folk medicines in some districts in China for relieving rheumatism and cold, regulating Qi (meridian) to alleviate pain. Previous studies revealed that this species contains major alkaloids, in specific aporphines, and minor flavonoids, lignans as well. Related pharmacological investigations demonstrated its activities and clinical applications on cardiovascular diseases, anti-cancer, against rheumatoid arthritis, relieving asthma and anti-allergic effects, as anti-oxidants, and so on. As an effort for further exploration of this bioactive ingredients and potential drug development, this paper summarizes most phytochemical and pharmacological results. Further, future prospects are also included.
Animals ; Drug Therapy ; Humans ; Litsea ; chemistry ; Molecular Structure ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry