Objective To study the mechanisms of extrarenal arterial blood supply of renal malignancy for its interventional therapy.Methods Routine abdominal aortography and selective questionable feeding arteriography were performed in 141 patients with renal malignancy.The characteristics and formation mechanisms of extrarenal arterial blood supply for renal malignancy were analyzed.Results Of the 141 patients,extrarenal arterial blood supply of renal malignancy were found in 51 patients and there were 87 branchs.The breakthrough of renal capsule with malignancy were found in those 51 patients.No extrarena]arterial blood supply of renal malignancy was found in 90 patients,including 50 patients with and 40 patients without the renal capsule breakthrough with malignancy.The emerge of extrarenal arterial blood supply of renal malignancy were significantly different(x~2 = 31.64,P

Objective To study the anti-tumor effects of humulon on arylamine N-acetyltransferase-1(NAT1)activity.Methods Employing HPLC,using PABA as substrate,in intact SGC-7901 cells and their cytoplasm,making PABA being acetylated to Ac-PABA by NAT1 as the activity of NAT1.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to study the expression of the NAT1 mRNA.Results The results show that humulon could inhibit the production of Ac-PABA in intact SGC-7901 cell and the cytoplasm,the production of Ac-PABA was gradually increased with the interaction time increasing.But comparing with corresponding negative control group's,the production of Ac-PABA was decreased evidently and the humulone could inhibit the expression of NAT1 mRNA.Conclusion Humulon could prevent the occurrence and deterioration of cancer.Its mechanisms can be attributed to its effect on decreasing the production of acetylation of carcinogenic aromatic amines,which is acetylated from aromatic amines,and inhibiting the NAT1 activity and expression of NAT1 mRNA.

Objective To explore the effects of humulon on kinetic parameters of N-acetyltransferase-1(NAT1) of human gastric cancer SGC-7901.Methods Employing HPLC,using para-aminobenzoic acid(PABA) as substrate,in intact SGC-7901 cells and their cytoplasm,taking the speed of PABA being acetylated to Ac-PABA by NAT1 as the rate of NAT1,using double reciprocal plot,taking the reciprocal of concentration of PABA and reaction rate of NAT1 as coordinates,regression equation was obtainied and the Michaelis constant(Km) and maximum reaction velocity(Vmax) were calculated.Results Study on enzyme kinetics demonstrated,as for intact SGC-7901 cells,Km and Vmax of control group were(3.910?0.087) ?mol/L and(0.306 0?0.006 7) pmol/L(1?106 cells),respectively,Km and Vmax of the humulon group were(3.830?0.123) ?mol/L and(0.275 0?0.005 8) pmol(1?106 cells),respectively.As for the cytoplasm of SGC-7901 cells,Km and Vmax of control group were(760.2?210.2) ?mol/L and(0.191 0?0.043 7) pmol/(mg?min),Km and Vmax of the humulon group were(449.0?72.9) ?mol/L and(0.094 0?0.010 4) pmol/(mg?min).Statistically,as for intact SGC-7901 cells or their cytoplasm,there was no difference of the Km between control group and humulon group,but there was remarkable difference of Vmax between control group and humulon group,P

OBJECTIVETo investigate the role of T cell and its subsets in the induction of insulitis and type 1 diabetes mellitus (T1DM) in BALB/c mice.

METHODSAutoimmune diabetes mellitus was developed by intraperitoneal injection of 40 mg/kg streptozotocin (STZ) daily for 5 consecutive days in BALB/c mice as sources of donor cells. Spleen cells from diabetic mice were then cultured for 7 days in the stimulation of interleukin-2 (IL-2) to harvest diabetogenic T cells, which were subsequently transferred into normal BALB/c mice recipients. MTT, ELISA, and HE staining were used to analyze the lymphocyte proliferation, cytokine (IL-2, interferon-gamma, IL-4, and IL-10) levels, and pathological changes in pancreatic islets.

RESULTSAs few as 3 x 10(6) diabetogenic T cells successfully induced diabetes mellitus in recipients pretreated with STZ twice, whereas transfer of equal amount of normal splenocytes, T cell-depleted diabetogenic splenocytes, or diabetogenic CD4+ T cells alone in recipients receiving STZ twice pretreatment was proved not to induce diabetes mellitus either. A markedly increased lymphocyte proliferation, high levels of interferon-gamma and IL-2 in the supernatants of diabetogenic T cells were observed. In addition, a markedly enhanced lymphocyte proliferation, a high level of interferon-gamma secretion in serum, and numerous lymphocytes infiltration in pancreatic islets were detected in the diabetic mice induced by diabetogenic T cells transfer.

CONCLUSIONSA novel T1DM murine model is established in STZ-pretreated BALB/c mice by adoptive transfer of diabetogenic T cells. CD4+ T cells with interferon-gamma may promote the onset of diabetes mellitus.

Animals ; Blood Glucose ; metabolism ; Cytokines ; immunology ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetes Mellitus, Type 1 ; immunology ; pathology ; Disease Models, Animal ; Islets of Langerhans ; cytology ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo develop a time-resolved fluoroimmunoassay (TRFIA) for detection of pertussis toxin (PT) S1 subunit for quality control of human PT vaccine.

METHODSA double antibody sandwich one-step method was used to establish the TRFIA for detecting PT S1 subunit in the vaccine.

RESULTSThe sensitivity of c peptide analysis reached 2.5 ng/ml without cross-reactions with other antigens. This assay could be used in detecting S1 subunit in the vaccine.

CONCLUSIONThe TRFIA for detecting PT S1 subunit is simple, sensitive and rapid for quality control of the PT vaccine.

Cross Reactions ; Fluoroimmunoassay ; methods ; Pertussis Toxin ; analysis ; Pertussis Vaccine ; chemistry ; standards ; Quality Control ; Sensitivity and Specificity

Heat-labile enterotoxin (LT) from Escherichia coli is a bacterial protein toxin with an AB5 hexamer structure. LT is a powerful mucosal adjuvant when co-administered with soluble antigens. However, its use in mucosal immunity is inconvenient because of its low yield and depolymerization during long-term storage under normal condition. In this study, we report an efficient expression system and optimized purification and storage strategy of LT. A gene encoding LT was cloned into the vector pET11c and transformed in E. coli BL21(DE3). By growing this strain on modified M9-CAA medium, LT was expressed efficiently. About 46mg/L LT could be purified from the supernatant of bacteria lysate. Using D(+)-Immobilized galactose column, LT could be purified at a wide pH range with various elution buffers. The optimized elution buffers are TEAN (pH 7.3) containing 0.3mol/L galactose and carbonate buffer (pH 10.4) containing 0.3mol/L galactose. After dried by freeze and placed in 4 degrees C, LT dissolved in TEAN (pH 7.3) and carbonate buffer (pH 10.4) were assayed by HPLC. The results indicated that the integrity of AB5 hexamer was kept well. LT could undergo long-term storage under this condition. This was proved to be an optimized strategy of LT storage. The results of GM1 binding assay and toxicity assay showed that the purified recombinant LT has normal biological character.
Animals ; Bacterial Toxins ; genetics ; isolation & purification ; metabolism ; pharmacology ; CHO Cells ; Cell Shape ; drug effects ; Chromatography, High Pressure Liquid ; Cricetinae ; Cricetulus ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; isolation & purification ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; isolation & purification ; metabolism ; pharmacology ; Female ; Genetic Vectors ; genetics ; Mice ; Mice, Inbred C57BL ; Toxicity Tests

BACKGROUNDDifferent strategies for hepatocellular carcinoma (HCC) may have distinct effects on the immune system. The aim of this research was to investigate changes in the immunological function after transcatheter arterial chemoembolization (TACE) plus radiofrequency ablation (RFA) in HCC patients.

METHODSA total of 51 consecutive HCC treatment-naïe patients was enrolled in this study and 20 healthy subjects served as controls. The therapeutic strategy was selected according to the tumor stage and general conditions. TACE was performed in 25 cases, TACE plus RFA in 17 and RFA in nine. All the patients underwent routine examinations and peripheral blood was harvested for the detection of lymphocyte subset by flow cytometry 1 day before, and 2 and 4 weeks after the treatment. The serum levels of alpha-fetoprotein (AFP), ALT and AST were also measured before and 4 weeks after treatment for the evaluation of therapeutic efficacy and liver function impairment.

RESULTSWhen compared with healthy controls, the CD4/CD8 ratio and the number of B cells and natural killer (NK) cells were significantly decreased in HCC patients before treatment (P < 0.05). When compared with before treatment, the CD4+ cells and CD4/CD8 ratio decreased but CD8+ cells increased in the TACE group (P < 0.05); the CD4/CD8 ratio and NK cells decreased but CD8+ cells increased in the TACE-RFA group (P < 0.05); the CD3+ cells, CD4+ cells, CD4/CD8 ratio and NK cells increased in the RFA group (P < 0.05). Significant differences in the CD3+ cells, CD8+ cells, CD4/CD8 ratio and NK cells were observed among groups (P < 0.05). Moreover, the AFP level decreased and transaminase level increased in all groups (P < 0.05). Differences of pre and post treatment between groups were statistically significant (P = 0.016, 0.025, 0.018 respectively).

CONCLUSIONSImmunity was compromised in HCC patients; TACE and TACE plus RFA lowered immunologic function to a certain extent. RFA improved it accompanied by a protective effect on liver function.

Adult ; Aged ; CD4-CD8 Ratio ; Carcinoma, Hepatocellular ; immunology ; therapy ; Catheter Ablation ; Chemoembolization, Therapeutic ; methods ; Combined Modality Therapy ; Female ; Humans ; Killer Cells, Natural ; immunology ; Liver Neoplasms ; immunology ; therapy ; Male ; Middle Aged ; alpha-Fetoproteins ; biosynthesis

As a kind of fluorescent nanoprobe, BHHCT-Eu3+ @ SiO2 fluorescent nanoparticles were synthesized using microwave irradiation. Transmission electron microscopy characterization showed that the nanoparticles were in spherical shape with particle size of about 36 nm. The BHHCT-Eu3+@SiO2 exhibited good fluorescence property, with a maximal excitation wavelength of 343 nm and an maximal emission wavelength of 615 nm. The fluorescence lateral flow immunoassay ( LFIA ) was established for rapid and quantitative detection of kanamycin ( Kana) in milk after BHHCT-Eu3+@SiO2 fluorescent nanoparticles were conjugated with Kana antibody, with dextran as a linker. The limit of detection of Kana with the LFIA method was 0. 85 ng/mL with IC50 of 12. 76 ng/mL, and the detection range was from 3. 0 ng/m to 76 ng/mL. The recoveries of Kana in milk were between 93 . 7% and 97 . 4% with RSD of 3 . 1%-4 . 6%, and cross-reactivity of the fluorescence immunoassay strip for Kana determination was<1%. The detection results of kana in milk samples between the LFIA and traditional ELISA method showed good correlation.

BACKGROUNDDrug treatment for secondary hyperparathyroidism caused by chronic renal failure may be available at the early stage of the disease, but it is not as effective for serious patients. The aim of the study was to evaluate the effect of total parathyroidectomy combined with forearm autotransplantation in the uremic patients with secondary hyperparathyroidism.

METHODSFrom September 1999 through September 2006, parathroidectomy and autotransplantation was performed in 20 patients. The coherence between the results of preoperative parathyroid ultrasonography and surgical exploration were compared. The serum calcium concentration and intact parathyroid hormone (iPTH) were monitored preoperatively, intraoperatively, and postoperatively.

RESULTSA total of 71 hyperplastic parathyroid glands were resected in the 20 patients. The accordance rate of parathyroid localization between B-ultrasonography and intraoperative exploration was 94.4%. The average iPTH value was (110.90 +/- 67.42) ng/L, (433.80 +/- 243.72) ng/L, (48.80 +/- 42.69) ng/L, (229.04 +/- 172.68) ng/L and (232.39 +/- 224.05) ng/L at day 1, 2, 3, 7, 30 after operation respectively. The clinical symptoms were ameliorated and the levels of serum calcium concentration were controlled within the normal range after operation. Recurrent secondary hyperparathyroidism had happened in 1 case, 4 years postoperatively because of the development of autograft hyperplasia, and in another case 2 years postoperatively due to remnant of neck parathyroid glands. The clinical symptoms were all alleviated after re-operation. No surgical complication had occurred in any of the patients.

CONCLUSIONSThe total parathyroidectomy with forearm autotransplantation is feasible, safe, and effective for patients with secondary hyperparathyroidism in the short term. The long-term effects should be further investigated.

Adult ; Aged ; Calcium ; blood ; Female ; Follow-Up Studies ; Forearm ; Humans ; Hyperparathyroidism, Secondary ; blood ; surgery ; Male ; Middle Aged ; Parathyroid Glands ; diagnostic imaging ; transplantation ; Parathyroid Hormone ; blood ; Parathyroidectomy ; methods ; Transplantation, Autologous ; Ultrasonography

OBJECTIVETo analyze the clinical factors affecting the recurrence of giant cell tumors (GCT) of bone.

METHODSThe complete data of 146 cases with GCT were reviewed. Thirteen clinical factors were analyzed by chi(2) analysis. And the related Campanacci's grade system and Jaffe's grade system was analyzed by Crosstabs analysis. Multipal factors were analyzed by Logistic regression analysis.

RESULTSNineteen of 146 cases recurred, and recurrence rate was 13.0%. Recurrence rates of curettage and enblock resection groups were 18.8% and 6.3% respectively. And recurrence rates of curettage with or without of extensive procedure were 12.9% and 38.9%. Five cases had lung metastasis, and two cases presented with malignant transformation. The metastasis rate and the rate of malignant transformation were 3.4% and 1.4% respectively. The two factors of surgery method and burst out of bone-envelope appearance were related with the recurrence. Moreover, Logistic regression revealed that the surgery method significantly affected the recurrence. And Campanacci's grade system and Jaffe's grade system were not related to each other.

CONCLUSIONSSurgery method is the main factor affected the recurrence of GCT, and Campanacci's grade system or Jaffe's grade system has no prognostic value.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; pathology ; surgery ; Factor Analysis, Statistical ; Female ; Giant Cell Tumor of Bone ; pathology ; surgery ; Humans ; Logistic Models ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Retrospective Studies ; Risk Factors