This study was aimed to investigate the immunological effect of modified dendritic cells (DC) which inducing cytotoxic T cells (CTL) against lymphoma cells. The DC were isolated from the lymph node and peripheral blood of patients with diffuse large B cell lymphoma (DLBCL). DC were transfected with recombinant adenovirus vector carrying human p53 gene (rAd-p53-DC). The expression of p53 gene was detected by flow cytometry. Western-blot was used to detect the expression of P53. ELISA was used to detect IL-12 level in supernatant. The mixed lymphocyte reaction (MLR) was used to detect the proliferative ability of auto-lymphocyte stimulated by DC. The lactate dehydrogenase (LDH) release test was used to determine the cytotoxicity of CTL. The results indicates that the expressions of DC surface molecule (except for CD1a) such as CD83, CD80, CD86 and HLA-DR were significantly higher in experiment group than that in control group and blank control group. The secretion of IL-12 in supernatant was higher in experiment group than that in control group. The autologous T lymphocyte proliferation and cytotoxic activity against the same kind of DLBL-cells increased in experiment group as compared with control group and blank control group (P < 0.05). The ability to stimulate T lymphocyte proliferation increased with the rising of the ratio of DC and T lymphocyte. However, there was statistically significant difference between rAd-p53-DC derived from Lymph node and peripheral blood (P < 0.05). It is concluded that rAd-p53-transfected DC can induce CTL response in vitro against lymphoma cells.
Adenoviridae ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; Genes, p53 ; Genetic Vectors ; Humans ; Lymphocyte Activation ; Lymphocyte Culture Test, Mixed ; Lymphoma, Large B-Cell, Diffuse ; blood ; immunology ; Transfection

Methylotrophic yeast, Pichia pastoris was used to express recombinant batroxobin, and a technology route of producing recombinant protein was finally established. We synthesized batroxobin gene artificially by means of recursive PCR. pPIC9-batroxobin was constructed and transformed into Pichia pastoris GS115 (his4). Recombinant batroxobin was expressed in yeast engineering strain and it was purified from the culture supernatant. 10 mg of recombinant batroxobin was purified from 1 liter fermentation media, it exhibited specific activity of 238 NIH units/mg and had molecular weight of 30.55 kD. The purified recombinant protein converted fibrinogen into fibrin clot in vitro, and shortened bleeding time in vivo. This study laid a foundation of development of hemostatic of recombinant snake venom thrombin-like enzyme.
Animals ; Batroxobin ; genetics ; metabolism ; pharmacology ; Electrophoresis, Polyacrylamide Gel ; Gene Expression ; Hemorrhage ; prevention & control ; Hydrogen-Ion Concentration ; Male ; Mice ; Pichia ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; metabolism ; pharmacology ; Time Factors

OBJECTIVETo compare clinical therapeutic effects of double moxibustion at Yifeng (TE 17) combined with electroacupuncture. and simple electroacupuncture on facial paralysis.

METHODSEighty-seven cases of facial paralysis were randomly divided into a treatment group (n = 47) treated by double moxibustion at Yifeng (TE 17) combined with electroacupuncture, and a control group (n = 40) treated by simple electroacupuncture. Their therapeutic effects were investigated after 30 days.

RESULTSThe total effective rate was 100.0% in treatment group and 87.5% in the control group with a significant difference between the two groups (P < 0.05).

CONCLUSIONDouble moxibustion at Yifeng (TE 17) combined with electroacupuncture in clinical therapeutic effect on peripheral facial paralysis is superior to simple electroacupuncture.

Acupuncture Points ; Electroacupuncture ; Facial Paralysis ; therapy ; Humans ; Moxibustion

Objective In this study,we investigated the relationship between oxidative stress,selenoproteins level and onset of Keshan disease (KD) through detecting the expression of 8-hydroxy-2-deoxy guanosine (8-OH-dG),thioredoxin reductase 1 (TrxR1) and glutathione peroxidase 1 (GPx1) in myocardial tissue.Methods Myocardium samples of autopsy patients including 8 cases of KD (KD group included 4 acute KD and 4 chronic KD) and 9 cases of non-KD (control group) were immunohistochemically stained for 8-OH-dG,TrxR1 and GPx1.The staining intensities subsequently quantified by using Olympus Image-Pro Plus 6.0 software.Results The positive rate of 8-OH-dG expression in myocardial nuclei was higher in the case group[(68.6 ± 20.4)％] than that of the control group[(2.4 ± 1.5)％,t =8.515,P ＜ 0.05].In addition,the positive rate of 8-OH-dG expression in acute KD[(91.7 ± 3.7)％] was significantly higher than that of chronic KD[(53.2 ± 7.9)％,t =6.409,P＜0.05].The distribution of TrxR1 and GPx1 was not associated with the distribution of myocardial damage.The expression of these two selenoproteins in KD group (401340 ± 59865,497590 ± 197082) were both lower than that of control group(2790300 ± 379298,1348400 ±615840; t =-28.493,-6.016,respectively,all P＜0.01).Conclusions Oxidative damage is detected in myocardium tissue of KD,and 8-OH-dG expression is associated with the degree of myocardial damage in KD.Selenoproteins,TrxR1 and GPx1,may be closely related to the pathogenesis of KD.

BACKGROUNDAnisodamine is widely used in therapy for treating acute glomerulonephritis and diabetic nephropathy because it can improve renal microcirculation. We performed a study to evaluate the preventive effects of anisodamine against contrast-induced nephropathy (CIN) in type 2 diabetics with renal insufficiency undergoing coronary angiography or angioplasty.

METHODSA total of 260 patients with type 2 diabetes and an estimated glomerular filtration rate (eGFR) of 60 ml(-1)×min(-1)×1.73 m(-2) or less, who were undergoing coronary angiography or angioplasty, were randomly assigned to receive an infusion of either sodium chloride (control group, n = 128) or anisodamine (treatment group, n = 132). Patients in the treatment group received an infusion of anisodamine at a rate of 0.2 µg×kg(-1)×min(-1) from 12 hours before to 12 hours after coronary angiography or angioplasty, while patients in the control group received an infusion of sodium chloride with the same volume as the treatment group. All patients received intravenous sodium chloride hydration. CIN was defined as a 25% increase in serum creatinine from baseline or an absolute increase of > 0.5 mg/dl within three days after contrast exposure. The primary end point was the incidence of CIN. The secondary end point was a 25% or greater reduction in eGFR.

RESULTSThere were no significant differences between the two groups with regard to age, gender, risk factors, laboratory results, medications and interventions. The incidence of CIN was 9.8% (13/132) in the treatment group and 20.3% (26/128) in the control group (P < 0.05). The secondary end point was 6.0% (8/132) in the treatment group and 16.4% (21/128) in the control group (P < 0.05).

CONCLUSIONThese results indicate the preventive effects of anisodamine against CIN in type 2 diabetics with renal insufficiency who are undergoing coronary angiography or angioplasty.

Acute Kidney Injury ; chemically induced ; prevention & control ; Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Contrast Media ; adverse effects ; Coronary Angiography ; adverse effects ; Creatinine ; blood ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Renal Insufficiency ; blood ; drug therapy ; Sodium Chloride ; administration & dosage ; Solanaceous Alkaloids ; therapeutic use

OBJECTIVESOrthotopic liver transplantation (OLT) is an accepted therapy for selected patients with advanced liver diseases. However, the early mortality rate after OLT remains relatively high due to the poor selection of candidates with various serious conditions. The aim of this study is to assess the value of pretransplantation artificial liver support treatment in reducing the pre-operation risk factors relating to early mortality after OLT.

METHODS50 adult patients in various stages of different etiologies who underwent OLT procedures had been treated with molecular adsorbent recycling system (MARS) preoperatively. The study was designed in two parts: the first one was to evaluate the effectiveness of a single MARS therapy by using some clinical and laboratory parameters which were supposed to be therapeutical pretransplantation risk factors. The second part was to study the patients undergoing OLT by using the regression analysis on preoperation risk factors relating to early (within 30 d after OLT) mortality rate.

RESULTSAmong the 50 patients, a statistically significant improvement of the biochemical parameters was observed (pretreatment vs posttreatment). 8 patients cancelled their scheduled LTXs due to significant improvements in their clinical conditions or recovery of their failing liver functions. 8 patients died and 34 patients successfully underwent LTX. The immediate outcome (within 30 postoperative days) of these 34 patients was that 28 were kept alive and 6 died.

CONCLUSIONSPreoperation sequential organ failure assessment (SOFA), level of creatinine, INR, TNFalpha, and IL-10 are the main preoperative risk factors relating to early death after an operation. MARS treatment before a transplant operation can relieve these factors significantly, hence improve survival rate of liver transplantation or even make the transplantation unnecessary.

Aged ; Factor Analysis, Statistical ; Female ; Humans ; Interleukin-10 ; blood ; Liver Cirrhosis ; surgery ; Liver Neoplasms ; surgery ; Liver Transplantation ; methods ; Liver, Artificial ; Male ; Middle Aged ; Preoperative Care ; Risk Factors ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo evaluate the influence of intracoronary administration of anisodamine on myocardial blush grade (MBG) and left ventricular regional and global systolic function and synchrony in the acute myocardial infarction (AMI) patients with no-reflow phenomenon post percutaneous coronary intervention (PCI).

METHODSForty-seven AMI patients who underwent PCI within 12 hours of onset and MBG was 0 - 1 were randomized to receive standard therapy [group B, n = 23, 18 males, mean age (62.72 +/- 11.48) years] or standard therapy plus intracoronary administration of anisodamine [200 microg/ml, group A, n = 24, 18 males, mean age (64.23 +/- 12.27) years]. The left ventriculography (LVG) was performed immediately and 6 months after PCI to measure the ventricular volume, LVEDP and wall motion score (WMS). Equilibrium radionuclide angiography (ERNA) was performed 1 week and 6 months after PCI to determine the parameters of left ventricular regional, global systolic function and systolic synchrony. Incidence of major adverse cardiac events (MACE) during the follow-up was analyzed.

RESULTSAnisodamine [(2530 +/- 340) microg/person)] was well tolerated by patients. The MBG remained unchanged in group B and significantly increased from grade 0.74 +/- 0.32 to grade 2.33 +/- 0.28 10 min after anisodamine injection in group B. Six months post PCI, LVESVI [(40.53 +/- 8.12) ml/m(2) vs. (50.32 +/- 8.26) ml/m(2)], LVEDVI [(80.13 +/- 9.74) ml/m(2) vs. (87.17 +/- 10.25) ml/m(2)], WMS [(8.24 +/- 1.31) vs. (10.23 +/- 1.82)] and LVEDP [(13.36 +/- 4.21) vs. (16.38 +/- 3.21) mm Hg, 1 mm Hg = 0.133 kPa] were significantly lower in group A compared with that in group B (all P < 0.05) while LVEF [(44.02 +/- 5.86)% vs. (38.52 +/- 5.18)%], PER [(1.86 +/- 0.09) EDV/s vs. (1.61 +/- 0.09) EDV/s] and PFR [(2.19 +/- 0.32) EDV/s vs. (1.78 +/- 0.17) EDV/s] measured by ERNA were significantly increased in group A compared with that in group B (all P < 0.05). (2) LrEF(2)-LrEF(8) in group A were higher by 13.96%, 25.02%, 30.36%, 22.86%, 27.67%, 22.07% and 18.71% respectively compared with that in group B. (3) Phase analysis showed that the left ventricular systolic synchrony parameters PS [(46.04 +/- 8.93) degrees vs. (53.19 +/- 162) degrees ], FWHM [(23.02 +/- 6.27) degrees vs. (25.02 +/- 5.31) degrees ] and PSD [(7.92 +/- 4.12) degrees vs. (11.76 +/- 4.11) degrees ] were also significantly lower in group A than that in group B (all P < 0.05). (4) During the 6 months of follow-up, the incidence of MACE in group A was significantly lower than that in group B (P < 0.05).

CONCLUSIONIntracoronary administration of anisodamine is safe and could partly attenuate the no-reflow phenomenon, improve the left ventricular systolic function and synchrony and reduce the incidence of MACE in patients with no-reflow phenomenon post AMI-PCI.

Aged ; Angioplasty, Balloon, Coronary ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; physiopathology ; therapy ; Myocardial Reperfusion ; Solanaceous Alkaloids ; administration & dosage ; therapeutic use ; Ventricular Function

Objective To explore the signal transduction mechanism of p38 mitogen activated protein kinase (p38MAPK) in human facial hypertrophic scar fibroblast (FB) differentiation into myofibroblasts(MFB).Methods Fibroblasts of primary culture were simple randomly assigned into two groups:cyclic stretch (control group) and cyclic stretch pre-treated with SB203580 (experimental group).Expression of P-p38MAPK and α-smooth muscle actin (α-SMA) protein were examined using Western blotting and expression of transforming growth factor β1 (TGF-β1) mRNA and α-SMA mRNA were examined using reverse transcription PCR(RT-PCR).Results In control group,the expressions of α-SMA,p38MAPK,TGF-β1 mRNA and α-SMA mRNA(0 h:0.134 ± 0.011,0.239 ± 0.015,0.214 ± 0.018,0.252 ± 0.010 ; 6 h:0.152±0.014,0.287 ±0.016,0.288 ±0.011,0.277 ±0.013; 12 h:0.172 ±0.017,0.320 ±0.017,0.335 ±0.013,0.297 ± 0.006),were significantly increased with loading time (6 h ＞ 0 h; 12 h ＞ 0 and 6 h).In experimental group (pre-treated with SB203580),the expressions of α-SMA,p38MAPK,TGF-β1 mRNA,α-SMA mRNA(6 h:0.116 ±0.017,0.128 ±0.016,0.134 ±0.014,0.163 ±0.009; 12 h:0.149 ± 0.013,0.136 ±0.018,0.144 ±0.013,0.187 ±0.010) on corresponding time decreased sharply compared with those in control groups (6,12 h).Conclusions The human facial hypertrophic scar fibroblasts differentiation in response to cyclic stretch was mediated by p38MAPK phosporylation.

BACKGROUNDPrevious studies have proved the renal protective effects of anisodamine in patients with septic shock. The aim of this study was to investigate anisodamine for the prevention of contrast induced nephropathy (CIN) in patients with acute coronary syndrome (ACS).

METHODSConsecutive ACS patients undergoing elective percutaneous coronary intervention (PCI) were randomly assigned to one of two groups: patients in the anisodamine group (ANI group) were assigned to receive intravenous infusions of anisodamine by an adjusted-dose (0.1 - 0.2 µg × kg(-1)× min(-1)) from the PCI procedure to 24 hours after PCI, and the control group (CON group) received 0.9% isotonic saline of the same volume. All patients were hydrated for 6 to 12 hours before and 12 hours after PCI. Blood samples were taken on the day of PCI and at 24, 48 and 72 hours after PCI to measure the serum creatinine (SCr).

RESULTSA total of 177 patients were involved in the study, 88 in the ANI group and 89 in the CON group. In both groups, the SCr concentrations significantly increased after PCI, with the peak value occurring at 48 hours. At 72 hours, the SCr concentration in the ANI group retuned to the baseline level (P > 0.05), but the SCr concentration in CON group was still higher than baseline level (P < 0.01). The SCr concentrations at 48 and 72 hours after PCI were much lower in the ANI group than those in the CON group (both P < 0.01). The estimated glomerular filtration rate (eGFR) significantly decreased after PCI, the lowest value occurred at 48 hours. In the ANI group, the eGFR at 72 hours was similar to the baseline level. In the CON group, the eGFR failed to return to baseline at 72 hours (P < 0.01). The eGFR at 24, 48 and 72 hours after PCI were higher in the ANI group (all P < 0.05). The incidence of CIN in the ANI group was lower than that in the CON group within 72 hours after PCI (P < 0.05). The results of multiple Logistic regression proved that both diabetes and left ventricular ejection fraction (LVEF) were independent predictors of CIN, and treatment with anisodamine was an independent preventive factor of CIN (OR 0.369 and 95%CI 0.171 to 0.794, P = 0.011). No serious side effects were found in the ANI group.

CONCLUSIONIntravenous infusion of anisodamine during and after elective PCI may safely prevent the occurrence of CIN in ACS patients.

Acute Coronary Syndrome ; therapy ; Adult ; Aged ; Angioplasty, Balloon, Coronary ; Contrast Media ; adverse effects ; Creatinine ; blood ; Female ; Glomerular Filtration Rate ; Humans ; Kidney Diseases ; chemically induced ; epidemiology ; prevention & control ; Logistic Models ; Male ; Middle Aged ; Solanaceous Alkaloids ; adverse effects ; therapeutic use

OBJECTIVETo compare the sensitivity and specificity of four kits for detection of anti-severe acute respiratory syndrome (SARS)-CoV IgG in sera of SARS patients.

METHODSAnti-SARS-CoV IgG was detected in 99 serial sera from 18 SARS patients and in 123 negative reference sera, using two enzyme linked immunosorbent assays (EIA No. A and No. B) and two indirect immunofluorescence assays (Australian IFA and Euroimmun IFA).

RESULTSAnti-SARS-CoV IgG was not detected in sera collected from SARS patients at the first week after onset by any of the four kits, however, it was detectable in sera obtained at the second week of illness by EIA No. B, and two IFA, but not by EIA No. A, with the positive rates of 57.1% (4/7), 57.1% (4/7) and 42.9% (3/7), respectively. The anti-SARS-CoV IgG was first determined in sera on the 9th day by Euroimmun IFA, 12th day by EIA No. B, 13th day by Australian IFA, and 16th day by EIA No. A. The positive rates of antibody on the 3rd week after onset were 84.2% (16/19), 94.7% (18/19), 78.9% (15/19) and 52.6% (10/19) respectively. They were identical since the 4th week after the disease onset. Through detection of 123 negative reference sera, the specificity of EIA No. A and two IFA was 100%, with exception of 94.9% for EIA No. B.

CONCLUSIONThe sensitivity and specificity of the two IFAs were relatively higher than that of the two EIAs. The quality of the two homemade EIAs should be improved.

Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoglobulin G ; blood ; Male ; Reagent Kits, Diagnostic ; SARS Virus ; immunology ; isolation & purification ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; diagnosis ; immunology ; virology