OBJECTIVETo establish the method to detect the cellular immune response of enhanced hepatitis B vaccine and make verification preliminary.

METHODSImmunized BALB/c mice with enhanced hepatitis B vaccine and detected the IFN-gamma spots forming cells (SFC) of mouse spleen cell by Elispot. Optimized the conditions of the experiment. Cellular immune response between enhanced hepatitis B vaccine and normal hepatitis B vaccine by Elispot were compared.

RESULTSIFN-gamma SFC was higher in 5microg dose than in 2microg dose after immunization with enhanced hepatitis B vaccine and IFN-gamma SFC was declined after immunization 3 weeks ago. IFN-gamma SFC was higher in stimulus by peptide than by protein. Compared to normal hepatitis B vaccine, IFN-gamma SFC was higher in enhanced hepatitis B vaccine.

CONCLUSIONEstablished the detection method to evaluate the cellular immunity of enhanced hepatitis B vaccine and tested the repeatability.

Animals ; Enzyme-Linked Immunospot Assay ; methods ; Female ; Hepatitis B ; immunology ; prevention & control ; virology ; Hepatitis B Surface Antigens ; administration & dosage ; immunology ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Humans ; Immunity, Cellular ; Interferon-gamma ; analysis ; immunology ; Mice ; Mice, Inbred BALB C

OBJECTIVETo explore the in vitro anti-tumor effect and mechanism of dendritic cell (DC) tumor vaccine induced by astragalus polysacharin (APS).

METHODSPeripheral blood mononuclear cells (PBMCs) isolated from human peripheral blood. DCs obtained from human peripheral blood were cultivated and added with culture solution for in vitro inducing them to immature DCs. On the 5th day of culture, 100 microg/mL (as the final concentration) APS was added to cells in the APS group. DCs were induced to mature in the cytokine groups by adding 20 ng/mL rhTNF-alpha (as the final concentration). Changes of morphology and phenotype of DCs were observed. Mature DCs were sensitized with tumor antigen SGC-7901 and co-cultured with allogeneic T cells. The proliferative function of T lymphocytes was detected by MTT assay. Levels of IL-12 and IFN-gamma in co-cultured supernatant were detected by ELISA. Cytotoxic lymphocytes (CTL) activated by DC were co-cultured with tumor cell SGC-7901. The specific killing capacity of CTL to target cells was detected by LDH release assay.

RESULTSThe morphological observation and phenotypic identification of APS induced DCs were in accordance with the characteristics of mature DCs. APS induced mature DCs could stimulate the proliferation of allogeneic T lymphocytes. The proliferation index of T cells increased with increased ratio of stimulator cells to effector cells (P < 0.05). Levels of IL-12 and IFN-gamma in co-culture supernatant significantly increased in a time-dependent manner (P < 0.05). CTL cells activated by sensitization of DCs could significantly kill tumor cells, and the killing effect increased along with increased effector-to-target ratio.

CONCLUSIONAPS could in vitro induce DCs to mature, promote its antigen-presenting capacity, effectively activate CTLs, and enhance anti-tumor function of the organism.

Antigen-Presenting Cells ; cytology ; drug effects ; immunology ; Cancer Vaccines ; immunology ; Cell Line ; Cell Proliferation ; drug effects ; Coculture Techniques ; Dendritic Cells ; cytology ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Interferon-gamma ; immunology ; Interleukin-12 ; immunology ; Leukocytes, Mononuclear ; cytology ; immunology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects

OBJECTIVETo observe the regulatory effect of assorted use of Chinese drugs for detoxifying and activating blood circulation on serum high sensitive C-reactive protein (hs-CRP) in apolipoprotein E gene knock-out [ApoE (-/-)] mice.

METHODSApoE (-/-) mice of 13-week old were devided into two groups, 12 in Group A fed with common forage and 98 in Group B with high lipid forage. Besides, 12 C57 BL/6J mice, 13-week old, were set as Group C fed with common forage. After being fed for 19 weeks, and the formation of vulnerable plaque had been confirmed in 2 mice of Group B, the other 96 mice were sub-divided into 8 groups, 12 in each group. Except that 0.4 mL normal saline was infused to the Group B1 as well as Group A and C, the other 7 sub-groups of Group B were respectively medicated with various drugs as follows: B2, Polydatin (PD, an extract from giant knotweed rhizome for detoxicating) 26. 6 mg/kg; B3, Xiongshao Capsule (XS, a Chinese herbal preparation for activating blood circulation), 110 mg/kg; B4, PD 53.2 mg/kg + XS 220 mg/kg; B5, PD 26. 6 mg/kg + XS 110 mg/kg; B6, PD 13.3 mg/kg + XS 55 mg/kg; B7, Lovastatin 3.3 mg/kg; and B8, Xuezhikang (XZK, a Chinese patent drug) 0.2 g/kg, all were administered by dissolving in 0.4 mL of distilled water for gastrogavage. The serum contents of hs-CRP were detected, with blood sample drawing from inferior vena cava, using enzyme-linked immunosorbent assay 17 weeks after medication.

RESULTSThe hs-CRP level was significantly higher in Group B1 than in Groups A and C (P <0.05, P <0.01). Comparisons between various sub-groups of Group B in hs-CRP content showed that hs-CRP in Group B2, B4 and B7 was lower than that in B1 (P <0.01), hs-CRP in Group B4 was the lowest, and hs-CRP was lower in Group B2 than in Group B3.

CONCLUSIONAssorted use of Chinese drugs for detoxifying and activating blood circulation could reduce serum hs-CRP level in ApoE (-/-) mice.

Animals ; Apolipoproteins E ; genetics ; metabolism ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; physiopathology ; Blood Circulation ; drug effects ; C-Reactive Protein ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Random Allocation

The diagnosis,surgical treatment,and comprehensive treatment of renal cell carcinoma with inferior vena cava tumor thrombus have advanced rapidly in recent years. Both the survival and quality of life of the patients have remarkably improved. Further advance in basic research may provide new direction of management of renal cell carcinoma.
Carcinoma, Renal Cell ; complications ; diagnosis ; therapy ; Embolism ; diagnosis ; etiology ; therapy ; Humans ; Kidney Neoplasms ; complications ; diagnosis ; therapy ; Venae Cavae

OBJECTIVETo evaluate the effect of transurethral holmium laser enucleation of the prostate (HoLEP) for prostate adenoma greater than 100 g.

METHODSSixty BPH patients with the prostate larger than 100 g were randomized to two treatment groups of HoLEP (n = 32) and open prostatectomy (n = 28). Comparisons were made between the two groups in operating time, blood loss, bladder irrigating time, catheterization time and hospital stay, as well as in the international prostate symptom score (IPSS), quality of life (QOL) score, maximum urinary flow rate (Qmax) and postvoid residual volume (PVR) before and 3 months after the surgery.

RESULTSCompared with the open prostatectomy group, the operating time was significantly longer (P < 0.01) but the blood loss, mean bladder irrigating time, catheterization time and hospital stay were significantly less in the HoLEP group (P < 0.01). Three-month follow-up revealed that HoLEP and open prostatectomy resulted in a similarly significant improvement in IPSS, QOL, Qmax and PVR (P < 0.01 ), with no statistical difference between the two groups (P > 0.05).

CONCLUSIONHoLEP and open prostatectomy are equally effective procedures for removal of large prostate adenomas, but the former is a better surgical option for prostate adenomas larger than 100 g for its greater safety, less pain and faster recovery.

Aged ; Holmium ; Humans ; Laser Therapy ; Male ; Prostate ; surgery ; Prostatectomy ; methods ; Prostatic Hyperplasia ; surgery ; Transurethral Resection of Prostate ; methods ; Treatment Outcome

AIMTo observe the effect of mesenteric lymph duct ligation on free radical and inflammatory mediator of myocardium with severe hemorrhagic shock in rats at different period, and explore the effect of intestinal lymphatic pathway on myocardium injury pathogenesis in shock rats.

METHODS78 male Wistar rats were divided into the sham group, shock group and ligation group. The model of serious hemorrhagic shock was established in shock group, ligation group, and mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group after resuscitate. All rats were executed and taken out heart making for homogenate of 10 percent to determine the MDA, SOD, tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL-6), myeloperoxidase (MPO), NO and NOS at after shock 90 min, after transfusion and resuscitate 0 h, 1 h, 3 h, 6 h, 12 h and 24 h etc. different times, and the expression of inducible nitric oxide synthase (iNOS) mRNA in myocardium was detected by RT-PCR.

RESULTSThe contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of shock group were rising after transfusion and resuscitate, and that was higher level at 3 h to 12 h, and that was significantly higher than sham group, the activity of SOD was significantly lower than sham group. The contents of MDA, TNFalpha, IL-6, MPO, NO, NOS and iNOS expression in myocardium of ligation group were significantly lower than that of shock group at sameness points, and the SOD activity was higher.

CONCLUSIONThe mesenteric lymph duct ligation and blocking mesenteric lymph could reduce the PMN detaining, decrease the discharging of TNFa and IL-6, reduce the NO and expression of iNOS mRNA, and reduce the releasing of free radical and consuming of SOD.

Animals ; Free Radicals ; metabolism ; Inflammation Mediators ; metabolism ; Interleukin-6 ; metabolism ; Lymphatic Vessels ; metabolism ; Male ; Mesentery ; metabolism ; Myocardium ; metabolism ; pathology ; Neutrophils ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; pathology ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate hepatitis E virus (HEV) infection among pigs in Henan province.

METHODSA total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively. The sera positive for HEV antigen were tested for HEV RNA with RT-PCR. The positive products of RT-PCR were cloned and sequenced.

RESULTSThe positive rates of anti-HEV antibody of the groups under 3-month and over 3-month of age were 90.27% and 92.55%, respectively, without statistical difference, while those of HEV antigen were 15.93% and 5.69%, respectively, with significant difference. The positive rates of anti-HEV antibody and HEV antigen were significantly different among different districts. HEV RNA was detectable in 5 of 47 HEV antigen positive samples. The sequence analysis showed that in 4 of 5 specimens the sequence belonged to genotype 4 while in the remaining one the sequence was genotype 1.

CONCLUSIONThe prevalence rate of HEV infection in pigs was high in Henan province and the rate differed in different districts.

Animals ; Antibodies, Viral ; analysis ; immunology ; Antigens, Viral ; analysis ; immunology ; China ; Genotype ; Hepatitis E ; epidemiology ; immunology ; veterinary ; virology ; Hepatitis E virus ; genetics ; immunology ; isolation & purification ; Phylogeny ; RNA, Viral ; analysis ; genetics ; Sequence Analysis, DNA ; Swine ; virology ; Swine Diseases ; epidemiology ; immunology ; virology

OBJECTIVETo investigate the possible protection provided by oral quercetin pretreatment against hepatic ischemia-reperfusion injury in rats.

METHODSThe quercetin (0.13 mmol/kg) was orally administrated in 50 min prior to hepatic ischemia-reperfusion injury. Ascorbic acid was also similarly administered. The hepatic content of quercetin was assayed by high performance liquid chromatography (HPLC). Plasma glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) activities and malondialdehyde (MDA) concentration were measured as markers of hepatic ischemia-reperfusion injury. Meanwhile, hepatic content of glutathione (GSH), activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and xanthine oxidase (XO), total antioxidant capacity (TAOC), contents of reactive oxygen species (ROS) and MDA, DNA fragmentation were also determined.

RESULTSHepatic content of quercetin after intragastric administration of quercetin was increased significantly. The increases in plasma GPT, GOT activities and MDA concentration after hepatic ischemia-reperfusion injury were reduced significantly by pretreatment with quercetin. Hepatic content of GSH and activities of SOD, GSH-Px and TAOC were restored remarkably while the ROS and MDA contents were significantly diminished by quercetin pretreatment after ischemia-reperfusion injury. However, quercetin pretreatment did not reduce significantly hepatic XO activity and DNA fragmentation. Ascorbic acid pretreatment had also protective effects against hepatic ischemia-reperfusion injury by restoring hepatic content of GSH, TAOC and diminishing ROS and MDA formation and DNA fragmentation.

CONCLUSIONIt is indicated that quercetin can protect the liver against ischemia-reperfusion injury after oral pretreatment and the underlying mechanism is associated with improved hepatic antioxidant capacity.

Administration, Oral ; Animals ; Antioxidants ; pharmacokinetics ; therapeutic use ; Ascorbic Acid ; pharmacokinetics ; therapeutic use ; Biological Availability ; Biomarkers ; blood ; DNA Fragmentation ; Glutathione Peroxidase ; metabolism ; Liver ; blood supply ; drug effects ; enzymology ; Male ; Malondialdehyde ; blood ; Quercetin ; pharmacokinetics ; therapeutic use ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Reperfusion Injury ; blood ; enzymology ; metabolism ; prevention & control ; Superoxide Dismutase ; metabolism ; Transaminases ; blood ; Xanthine Oxidase ; metabolism

Translocating protein and translocating peptides have therapeutic potential against tumors by exposing the cytotoxic domains of toxic proteins to the cell cytosol. The aim of this study is to investigate the effect of N-terminally fused PE translocating peptides on granzyme B (GrBa) activity. PE II-GrBa fusion protein genes were constructed by replacing N-terminal signal and acidic dipeptide sequence of human granzyme B gene with two truncated translocating sequences of Pseudomonas exotoxin A (PE II aa 280-364/358) by recombinant PCR, and then cloned into pIND inducible expression vector. The resulting pIND-PE II-GrBa expression vectors were co-transfected with assistant plasmid pVgRXR into HeLa cells through lipofectamine, followed by selection on G418 and zeocin. The resistant cells were collected and induced with ponasterone A. Western blot analysis demonstrated that ponasterone A induction caused the expression of PE II-GrBa fusion proteins, and indirect immunofluorescence detected giant sized multinucleated cells, suggesting cytoskeletal and mitotic abnormalities as reported in our previous studies. Western blot, enzymatic activity assay and cell counting analysis indicated that two types of PE II-GrBa fusion proteins were capable of cleaving both endogenous and exogenous substrates of granzyme B, and inhibiting the growth of cells. The PE II (aa 280-358)-GrBa was shown to have higher serine protease activity and stronger growth inhibitory effect. Such inhibition was presumably associated with G2 arrest as determined by cell cycle analysis. These data prove that PE II-GrBa fusion proteins have cell inhibitory effect similar to GrBa, and that the shorter PE-derived peptide exerts less influence on GrBa activity. This study helps to optimize the construction of recombinant protein comprising translocating peptides and cytotoxic molecules for tumor cell killing.
ADP Ribose Transferases ; genetics ; pharmacology ; Bacterial Toxins ; genetics ; pharmacology ; Cell Proliferation ; drug effects ; Exotoxins ; genetics ; pharmacology ; Granzymes ; genetics ; pharmacology ; HeLa Cells ; Humans ; Recombinant Fusion Proteins ; pharmacology ; Virulence Factors ; genetics ; pharmacology

OBJECTIVETo study the effect of hBcl-2 gene transfer on rat liver against ischemia-reperfusion injury, and explore the feasibility of this approach to reduce ischemia-reperfusion injury in liver transplantation.

METHODSWe constructed the replication-deficient recombinant adenoviruses Adv-EGFP and Adv-Bcl-2 and transfected them into 293 cells and packaged into adenovirus particles for amplification and purification. The empty plasmid vector virus was constructed similarly. Male SD rats were randomized into Adv-Bcl-2-transfected group, Adv-EGFP-transfected group, ischemia-reperfusion group, and sham-operated group, and liver allograft transplantation model was established by sleeve method. In the transfected groups, the recombinant viruses were administered by perfusion through the portal vein, and the ischemia-reperfusion and sham-operated groups received no treatment. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of bcl-2 in the liver tissue of each group, and at 0, 60 and 180 min after reperfusion, serum AST, LDH, and MDA levels were measured. Histological changes of the liver cells were evaluated by HE staining.

RESULTSBcl-2 mRNA and protein expressions in Adv-Bcl-2-transfected group, as compared with those in Adv-EGFP-transfected group and control group, were significantly increased (P<0.01); the serum levels of AST, LDH and MDA in Adv-Bcl-2-transfected group were significantly lower than those of Adv-EGFP-transfected group and ischemia-reperfusion group (P<0.05 or 0.01). Compared with the sham-operated group, Adv-Bcl-2 treatment group showed lessened edema and vacuolar degeneration of the liver cells without patches or spots of necrosis. In ischemia-reperfusion and Adv-EGFP group, HE staining revealed hepatic lobular destruction and extensive liver cell swelling, enlargement, vacuolar degeneration, edema and occasional focal necrosis.

CONCLUSIONAdv-Bcl-2 transfection can induce the expression of bcl-2 gene to reduce ischemia-reperfusion injury of the liver graft in rats.

Animals ; Genes, bcl-2 ; Liver ; blood supply ; Liver Transplantation ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; prevention & control ; Transfection