Objective To compare the potential cytotoxicity induced by Veratrum nigrum coadministered with Panax ginseng, and to provide experimental evidence on the mode of herb-herb interaction based on human liver drug metabolizing enzymes.Methods The effect of V.nigrum and coadministration on cultured human hepatoma (HepG2) cells was investi-gated by detecting morphological changes , cell viability , cytomembrane integrity and apoptosis after the cells were treated for 24 h.The mRNA expression levels of drug metabolizing enzymes influenced by P.ginseng were determined by real-time quantitative reverse-transcriptase polymerase chain reaction .Results V.nigrum coadministered with P.ginseng had a better inhibitive effect on the growth of HepG2 cells at the IC50value of (15.18 ±1.03) mg/ml than at the value of IC50 (21.46 ±1.10) mg/ml of V.nigrum.Coadministration more significantly raised the LDH level in cell culture medium than at the same dose of V.nigrum.Moreover, in coadministration group, compared with the same dose of V.nigrum,the total apoptosis and necrosis of HepG2 cells were significantly increased .P.ginseng had effect on the expression of CYP3A4, CYP1A1, CYP1A2, CYP2B6 and CYP2E1 mRNA.Conclution Compatibility of medicines in a prescription also has herb-herb interactions based on drug metabolizing enzymes .The interaction mode is that the P.ginseng inhibits and induces CYPs and the modulated CYP isozymes ,inturn,have an impact on the metabolism of constituens in coadministered herbs causing herb-herb interaction .

Objective To evaluate the regulatory effect of OX40 co-stimulatory signal on the expression of Foxp3 in inductive regulatory T cells (iTreg) in vitro.Method CD4+ CD25+ naive T cells were isolated from C57BL/6 mouse lymphocyte suspension by MASC CD4+ CD25+ regulatory T cell isolation kit.Inductive Tregs were generated by stimulation of naive T cells in the presence of transforming growth factor beta (TGFβ1),anti-CD3,anti-CD28 and IL-2.The regulatory effect on iTregs was shown by use of OX40 stimulation monoclonal antibody (OX86) or control antibody.Using flow cytometric analysis (FACS),we examined the antibody-based identification of Tregs surface markers CD4 and CD25,along with the intracellular activation marker FoxP3.Results The ratio of CD4+ CD25+ nTregs isolated from mouse lymphatic node was (5.0 ± 0.4)％ vs.(71.8 ± 13.4)％ of TGFβ1-driven iTregs.The ratio of CD4+ CD25+ Tregs was (80.0 ± 1.6) ％ in OX40 stimulation McAb group vs.(86.0 ± 1.4)％ in control antibody group.Furthermore,the expression of Foxp3 was (59.2 ± 0.7) ％ in OX40 stimulation McAb group vs.(70.0 ± 0.8) ％ in control antibody group (P＜0.05).Conclusion TGFβ1-dependent protocol may induce the conversion of naive CD4+ T cells into CD25+ Foxp3+ iTregs.OX40 stimulation can down-regulate the expression of Foxp3 in CD4+ CD25 + iTreg significantly.Thus OX40 molecular may become an attractive target in Tregs-induced transplant tolerance.Further study should be performed to increase the suppressive activity of iTregs through blockade of OX40 signal.

Action Potentials ; drug effects ; physiology ; Animals ; Anti-Arrhythmia Agents ; pharmacology ; Female ; Heart Ventricles ; metabolism ; Isoflavones ; pharmacology ; Male ; Microelectrodes ; Myocardium ; metabolism ; Patch-Clamp Techniques ; Potassium Channels, Inwardly Rectifying ; metabolism ; physiology ; Rats ; Ventricular Function ; physiology

Objective To investigate whether rat adipose-derived stem cells (rASCs) could resist human xenoantibody-dependent complement-mediated lysis and to explore its possible mechanisms.Method SD rat ASCs were isolated,rASCs at passage 2 to 8 were used for the following studies and rat lymphocytes were harvested as control cells.α-Gal expression was detected by flow cytometry.After incubation of rASCs with 20％ normal human serum (NHS) or heat inactivated normal human serum (HINHS),flow cytometry was used to detect cytotoxicity,IgG or IgM binding,and C3c,C4c and C5b-9 deposition.Result We successfully established the method to isolate and culture rASCs.The morphology of rASCs remained unchanged after passages.rASCs were positive for tell surface markers of CD44 and CD90,while negative for CD45 and MHC-Ⅱ.As compared with rLCs,rASCs significantly resisted human natural antibody and complement-mediated lysis when incubated with 20％ NHS in vitro (20.42％ ± 2.80％ vs 51.84％ ± 6.70％,P ＜ 0.01).Mechanistically,rASCs expressed lower level of α-Gal (13.97 ± 0.33 vs.24.47 ± 3.03,P＜0.05),which was correlated with decreased binding of human xenoreactive IgG and IgM (IgM:9.4 ± 2.0 vs.107.2± 4.8,P＜0.01; IgG:5.73 ± 1.0 vs.27.49 ± 3.9,P＜0.01) and reduced deposition of complements C3c,C4c and C5b-9 (C3c:294.6 ± 38.02 vs.1924 ± 509.4,P＜0.05; C4c:35.23 ± 3.1vs.177.3 ± 37.17,P＜0.05; C5b-9:5.63 ± 1.74 vs.37.05 ± 7.4,P＜0.01).Conclusion These data demonstrated that the resistance of rASCs to human xenoantibody and complement-mediated lysis is associated with low expression of xenoantigen a-Gal and inhibition of MAC (membrane attack complex) formation.

Objective To investigate the clinical significance of retinal binding protein (RBP) and Cystatin C in blood and urine in patients with chronic kidney disease (CKD).Methods One hundred and twenty-one patients with CKD in Nephrology Laboratory of The People's Hospital of Xinjiang Uygur Autonomous Region from Aug.2012 to Jan.2013 were served as the case group.Sixty healthy check-up people were considered as control group.Enzyme linked immunosorbent assay was used to detect RBP and Cystatin C in urine,and the immune turbidimetry testing was applied to detect RBP and Cystatin C in blood.The receiveroperating characteristic (ROC) curve and area under curve (AUC) were applied to evaluate the sensibility and diagnostic value of indexes in CKD.Results The positive rate of RBP,Cystatin C in urine and blood were 89.6％,92.6％,52.5％,59.5％ respectively of 12 patients in case group,higher than those in control group (all value were 0,P ＜0.001).ROC curve showed that the sensitivity of urine RBP and Cystatin C were higher than that in blood of case group.AUC of RBP,Cystatin C level in urine and blood were 0.915,0.974,.655,and 0.623 respectively.And 95％ confidence interval were 0.877-0.954,0.956-0.992,0.575-0.736,0.543-0.702 respectively,and the differences are statistically significant (P ＜ 0.001).Conclusion The sensitivity of urine cystatin C and RBP are significantly higher than that in blood,which might be served as a diagnostic marker of CKD and can provide important basis for clinical diagnosis.

10.3969/j.issn.1008-9691.2013.03.011

Objective To explore the differentiation of allogeneic naive T cells to regulatory T cells (Tregs) and T helper (TH) 1/2/17 cells by coculture with bone marrow-derivedimmature dendritic cells (irnDC).Method Bone marrow-derived imDC were cultivated from Balb/c mice.Lipopolysaccharide-stimulated DC were harvested as mature dendritic cells (mDC) and unstimulated cells were collected as imDC.Then irnDC or mDC were cocultured with allogeniec naive T cells,respectively.TH1 cytokines [interferon-γ (IFN-γ) and interleukin (IL)-2],TH2 cytokines (IL-4 and IL-10),and TH17 cytokine (IL-17) of co-cultured cells were detected by enzyme linked immunospot assay.CD4+ Forkhead box p3 (FoxP3) + Treg proportion in CD4+ cells in the co-cultured system with IL-2 and transforming growth factor-β1 (TGF-β1) was analyzed by flow cytometry.Result As compared with mDC,na(i)ve T cells cocultured with imDC secreted much less IFN-γ (11.67 ± 2.18 vs.182.00±23.71,P＜0.01),IL-2 (26.67±2.96 vs.318.30± 18.62,P＜0.01),IL-4 (17.00±3.78 vs.45.33±3.48,P＜0.01),IL-10(7.00±1.00vs.158.70±10.90,P＜0.001) and IL-17 (0.66 ± 0.33 vs.238.30 ± 24.39,P＜0.001).Furthermore,imDC induced more CD4+ FoxP3+ Tregs than mDC after adding IL-2 and TGF-β1 in the coculture system for 7 days (22.70 ± 1.53 ％ vs.5.42 ± 1.27％,P＜0.01).Conclusion imDC are more effective to induce na ve T cells to Tregs,but not differentiate to TH 1/TH 2/TH 17 cells.These findings provide in vitro experimental evidence for induction of transplant tolerance by adoptive transfer of imDC.

Objective To investigate the treatment and prevention strategies of hematomas in operation area after anterior approach surgery for cervical spondylosis.Methods A retrospective review was conducted on 12 with hematoma compression in operation area out of 785 patients managed by anterior cervical surgery from January 2007 to July 2013,including 10 males and 2 females at age ranging from 40-71 years (mean 56.8 years).Surgery method was anterior cervical corpectomy and interbody fusion using titanium mesh cage plus plate and intraoperative blood loss was 300-1 200 ml.Primary clinical manifestations were neurological dysfunction in 5 patients,dyspnea in 6,and both neurological dysfunction and dyspnea in 1.There were 10 patients with the presence of symptoms at postoperative 0.5-22 hours,1 at postoperative 73 hours,and 1 at postoperative 74 hours.All the 12 patients underwent a second anterior cervical exploration.Results There were 5 patients with epidural hematoma,6 with subcutaneous hematoma,and 1 with both hematomas.After surgical interventions,the patients presented improvement in respiratory and neurological function,with inapparent respiratory abnormality and improved neurological function at discharge.One patient was died of cardiovascular-associated disease after being discharged from hospital.The left 11 patients were followed up for mean 19.8 months (range,6-43 months),with improved Japanese Orthopedic Association (JOA) score at final follow-up.Conclusions Hematoma took place frequently in the early period,especially within 24 hours in operation area after anterior approach to cervical disorders and close attention should be paid to respiratory and limb sensation and motion functions.Early detection and early surgical interventions are the key countermeasures to avoiding the severe results.

OBJECTIVETo investigate the blood quality of blood banks.

METHODSHBsAg, Anti-HCV, Anti-HIV were detected by enzyme linked immunoadsorbent assay (ELISA). Treponema pallidum was detected by Trust or rapid plasma regain card test (RPR) before August 2001 and ELISA was used afterwards. Alanine aminotransferase (ALT) was detected by Reitman-Frankel before December 1999 and by continuous monitoring assay afterwards. Results were compared between (1) sampling and spot check, (2) different determent methods, (3) two kinds of reagents with different antigen coating.

RESULTSSeventy-two (8.28 per thousand ) of the 8,699 plasma samples were found unqualified with levels of anti-HCV, ALT, HBsAg, Treponema pallidum and anti-HIV 24 (2.76 per thousand ), 19 (2.18 per thousand ), 16 (1.84 per thousand ), 7 (0.80 per thousand ), 6 (0.69 per thousand ), respectively. There were significant differences between different determent methods and different reagents, but only ALT showed significant difference in the sampling and spot check.

CONCLUSIONThe unqualified samples were associated with testing methods, quality of reagents as well as ability and responsibility of the staff.

Alanine Transaminase ; blood ; Blood Banks ; standards ; Blood Donors ; Blood-Borne Pathogens ; isolation & purification ; China ; D-Alanine Transaminase ; Enzyme-Linked Immunosorbent Assay ; HIV Antibodies ; blood ; HIV Seropositivity ; Hepatitis B Surface Antigens ; blood ; Hepatitis C Antibodies ; blood ; Humans ; Quality Control

In order to investigate the characteristics of transdermal delivery of ferulic acid under the treated of microneedle arrays and the influence on permeability of rat skin capillaries, improved Franz-cells were used in the transdermal delivery experiment with the rat skin of abdominal wall and the length of microneedle arrays, different insertion forces, retention time were studied in the influence of characteristics of transdermal delivery of FA. The amount of FA was determined by HPLC system. Intravenous injection Evans blue and FA was added after microneedle arrays treated. Established inflammation model was built by daubing dimethylbenzene. The amount of Evans blue in the rat skin was read at 590 nm wavelength with a Multiskan Go microplate reader. Compared with passive diffusion group the skin pretreated with microneedle arrays had a remarkable enhancement of FA transport (P <0.01). The accumulation of FA increased with the enhancement of insertion force as to as the increase of retention time. Microneedle arrays with different length had a remarkable enhancement of FA transport, but was not related to the increase of the length. The research of FA on the reduce of permeability of rat skin capillaries indicated that the skin pretreated with microneedle arrays could reduce the content of Evans blue in the skins of rat significantly compared with the untreated group. The permeation rate of ferulic acid transdermal delivery had remarkable increase under the treated of microneedle arrays and the length of microneedle arrays ,the retention time so as to the insertion force were important to the transdermal delivery of ferulic acid.
Administration, Cutaneous ; Animals ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Skin Absorption