Objective To construct a recombinant bacillus Calmette-Guérin(BCG) vaccines based on different tandem repeats of MUC1 and GM-CSF, rBCG-MVNTR1/4/8-CSF, and to observe the ability of three recombinant BCG vaccines in the inhibition of breast cancer. Methods After MUC1 variable-number tandem repeats (MVNTR1/4/8) were cloned in a stepwise manner, the E. coli-Mycobacteria shuttle expression vector pDE22-MVNTR1/4/8-CSF were constructed by fusing MVNTR1/4/8 and GM-CSF, and then used to transform competent BCG by electroporation after identification by restriction endonuclease digestion analysis and DNA sequencing. A novel breast cancer vaccines, rBCG-MVNTR1/4/8-CSF was constructed. The expression of fused MVNTR1/4/8-CSF protiens was analyzed by SDS-PAGE and Western blot. The ability of rBCG vaccines inhibiting the growth of breast cancer was observed in hu-PBL-SCID mice. The specific T cell responses in mice were assessed by immunohistochemistry. Results The expression of recombinant MVNTR1/4/8-CSF fusion proteins were detected by SDS-PAGE and Western Blot in rBCG-MVNTR1/4/8-CSF vaccines, respectively. Tumor incidence in mice prophylactic immunized with rBCG-MVNTR4-CSF (37.5%) or rBCG-MVNTR8-CSF (25%) significantly decreased compared with PBS and BCG-pDE22 control ( 100% ) at 42 days after tumor implantation ( P ＜ 0. 05 ). MCF-7 tumor growth inhibition in rBCG- MVNTR4/8-CSF-immunized mice was more significant than that in controls ( P ＜0. 01 ).The inhibition effect of three rBCG vaccines on breast rumor growth appeared to rise with increase of numbers of the tandem repeats of MUC1. Survival rate was 75% of mice in the rBCG-MVNTR4-CSF-treated group and 87. 5% of mice in the rBCG-MVNTR8-CSF-treated groups at 70 days after tumor implantation; however,survival rate was only 12. 5% in control group( P ＜0. 05). The CD4-positive and CD8-positive lymphocytes were detected only in rBCG-MVNTR4/8-CSF-immunized mice. Conclusions rBCG-MVNTR4/8-CSF immunization inhibits breast cancer growth in mice.

AIM To compare the contents of benzoyl mesaconitine,benzoyl hypaconitine,benzoyl aconitine,aconitine,hypaconitine and mesaconitine in Zingiberis Rhizoma Recens boiled juice-processed,Zingiberis Rhizoma boiled juice-processed,Zingiberis Rhizoma Recens juicing-processed,Zingiberis Rhizoma Recens slice malaxation steaming,and Zingiberis Rhizoma slice malaxation steaming Aconiti lateralis Radix Praeparata.METHODS Acid-base titration method was adopted in the content determination of total alkaloids.HPLC was applied to determining the contents of six alkaloids,the analysis was performed on a 35 ℃ thermostatic Waters Symmetry(C)C1scolumn (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.04 mol/L ammonium acetate flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 235 nm.RESULTS The contents of total alkaloids and diester-type alkaloids (mesaconitine,aconitine and hypaconitine)in various processed products,especially for Zingiberis Rhizoma Recens slice,Zingiberis Rhizoma slice malaxation steaming products,were obviously lower than those in the raw product.The contents of benzoyl hypaconitine and benzoyl aconitine in Zingiberis Rhizoma slice malaxation steaming product were the lowest,while that of benzoyl mesaconitine in the raw product was the lowest.CONCLUSION Compared with other Zingiber-processing methods,both Zingiberis Rhizoma Recens slice and Zingiberis Rhizoma slice malaxation steaming can reduce the toxicity of Aconiti lateralis Radix Praeparata more effectively.

Aim To investigate the molecular mecha-nism for the inhibitory effect of puerarin on IL-6 secre-tion in human osteoblast-likeMG-63 cells. Methods According to our previous studies, human osteoblast-like MG-63 cells containing two estrogen receptor ( ER) isoforms are a suitable model for this study. En-zyme-linked immnosorbent assay (ELISA), RT-PCR, luciferase reporter assay and small interfering RNA were performed to investigate the effect of puerarin on IL-6 expression in osteoblast-derived cells and underly-ing molecular mechanism. Results Puerarin could obviously inhibit IL-6 expression and IL-6 promoter ac-tivity by human osteoblastic MG-63 cells. Treatment with the ER antagonist ICI 182,780 abrogates the a-bove actions of puerarin on osteoblast-derived cells. U-sing siRNA technology, we further demonstrated that the effects of puerarin on IL-6 production were media-ted by ERα. Conclusion The effect of puerarin on IL-6 production in osteoblast is mediated by ERα.

Objective To investigate the effects of enteral nutrition with galactooligosaccharides (GOS) on serum inflammatory cytokines in rats with severe acute pancreatitis (SAP). Methods SD rats were randomly divided into sham operation control group, SAP with enteral nutrition (EN) group and SAP with EN supplemented with GOS (GOS-EN) group, and each group was divided into 4 d and 7 d subgroups according to the time that animals were sacrificed (n=8 in each subgroup). Rat SAP models were established by injection of 38 g/L sodium taurocholate beneath the pancreatic capsule. The serum amylase, inflammatory cytokines TNF-α, IL-2 and IL-10 were detected. Results At each time point, the levels of serum amylase in all SAP groups were significantly higher than those in sham operation control group (P < 0.01), and the levels in GOS-EN group were significantly lower than those in EN group (P < 0.01). The levels of serum TNF-α and IL-10 in all SAP groups were significantly higher than those in sham operation control group (P < 0.01), while the levels of IL-2 in all SAP groups were significantly lower than those in sham operation control group. The levels of TNF-α in GOS-EN group were significantly lower than those in EN group (P <0.05), while the levels of IL-2 and IL-10 and the ratio of IL-10/TNF-α were significantly higher than those in GOS-EN group (P < 0.05). Conclusion Early EN supplemented with GOS could modulate the balance of pro- and anti-inflammatory response.

ObjectiveTo investigate the iodine nutritional status of residents in Shanxi province,and to provide a scientific basis for adjustment of control strategies and measures to iodine deficiency disorders (IDD).MethodsIn the 11 cities and 119 counties(cities,districts),except high water iodine townships,9 townships were selected in each county according to their sub-area positions of east,west,south,north and center,4 villages were sampled in each chosen township,and 8 households were selected in each chosen village in every chosen county (cities,districts ) with 9 or more townships.In every chosen county (cities,districts) with 6 to 9 townships,1 township was selected respectively in east,west,south,north and center sub-areas of the township,4 villages were sampled in each chosen township,and 15 households were selected in each chosen village.In the county (cities,districts) with 5 or less townships,all township were selected,4 villages were sampled in each chosen township,and 15 households were selected in each chosen village.Edible salt samples from these households were collected; iodized salt was determined by direct titration.In the 119 counties(cities,districts),1 township was selected,respectively,in east,west,south,north and center sub-areas in each county,and 20 children aged 8 - 10 in each of the selected townships were selected to collect urine samples and urinary iodine was determined by As-Ce catalytic spectrophotometry.Evaluation criteria:median urinary iodine ＜ 100 μg/L was iodine deficiency,100 - 199 μg/L as appropriate,200 - 299 μg/L as more than appropriate,and ≥ 300 μg/L as iodine excess.ResultsMedian iodine of the 34 808 household salt samples was 31.55 mg/kg.The coverage rate of qualified iodized salt was 99.18％(34 521/34 808) and the consumption rate of qualified iodized salt was 97.12％(33 805/ 34 808).In the 11 cities,119 counties(cities,districts),the median of urinary iodine of 11 967 children aged 8 -10 was 244.0 μg/L,of which ＜ 50 μg/L acoounted for 2.6％(312/11 967),50 - 99 μg/L accounted for 6.9％(823/11 967),100- 199 μg/L accounted for 26.3％(3145/11 967),200 - 299 μg/L accounted for 28.7％(3440/11 967),and 300 μg/L or higher accounted for 35.5％(4247/11 967).The medians of urinary iodine in the 9 municipal cities were 200 - 300 μg/L,and other 2 cities were 300 - 400 μg/L At the county level,the medians urinary iodine of children of the 119 counties(cities,districts) were 100 - 199 μg/L that accounted for 15.1％(18/119),200 - 299 μg/L accounted for 63.9％(76/119),and 300 μg/L or higher accounted for 21％(25/119).Conclusions The iodine nutrition level of residents in Shanxi province is more than appropriate.The salt iodine concentration in Shanxi province needs to be reduced,but the space is not wide.

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

Objective To study the sources and the relationship between the management and the outcome of hemorrhage after cephalic pancreatoduodenectomy.Methods The clinical data of 370 patients who underwent pancreatic resection at the Lihuili Hospital and the Second Affiliated Hospital of Zhejiang University were retrospectively analyzed.Results Postoperative bleeding occurred in 35 patients with 11 deaths.Among those intraabominal bleeding occurred in 14 cases and gastrointestinal hemorrhage occurred in 22,with one case suffering from both.Bleediug developing within 72 hours after operation in 12 cases (early-stage group),which was caused by improper intraoperative homeostasis.In other 23 cases,bleeding 72 hours after operation(later stage group)was caused by the erosion following pancreatic and/or bile leakage.Relaparotomy was performed in 13 cases and endoscopic homeostasis was performed in 3. Relaparotomy or endoscopic homeostasis was superior to that of conservative therapy in the early-stage group (P0.05).Pancreatic or bile leakage was identified as the significant risk factors for the postoperative bleeding.Conclusions In order to prevent the postoperative hemorrhage and to reduce the mortality of pancreatic resection,skillful techniques,expeditious homeostasis,proper management of stump pancreas and the prevention of pancreatic and bile leakage are essential.

OBJECTIVETo explore the effect and mechanism of Poly I:C in inducing growth inhibition and apoptosis of human hepatocellular carcinoma SMMC-7721 cells.

METHODSSMMC-7721 cells were treated with different doses of Poly I:C for 24, 48, and 72 h, and the cell growth inhibition rate was analyzed with CCK-8 assay. The cell cycle and the apoptosis were analyzed using flow cytometry with Annexin-V and PI staining, and quantitative RT-PCR analysis were used to detect the expression of TLR3, TRIF, and IFN-beta mRNA in cells.

RESULTSIn the cells exposed to Poly I:C at low, moderate, and high doses, the inhibitory rates was the highest in high-dose Poly I:C group, and at a given Poly I:C dose, prolonged exposure resulted in significantly increased cell growth inhibition rate (P<0.05). Flow cytometry showed that Poly I:C induced cell apoptosis in a time- and dose-dependent manner and significantly increased the percentage of G1-phase cells as compared with that in the control group. The mRNA level of TLR3, TRIF, and IFN-beta were also increased following Poly I:C treatment in comparison with the control group.

CONCLUSIONPoly I:C can induce significant growth inhibition and apoptosis of SMMC-7721 cells in a dose- and time-dependent manner possibly by causing cell cycle arrest and TLR3 signaling pathway activation that leads to IFN-beta production and cell apoptosis.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Interferon-beta ; genetics ; metabolism ; Liver Neoplasms ; pathology ; Poly I-C ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Receptors, Cholecystokinin ; metabolism ; Signal Transduction ; Toll-Like Receptor 3 ; genetics ; metabolism

OBJECTIVETo investigate the inhibitor of matrix metalloproteinase-2 (MMP-2) (2R)-2-[5-[4-[ ethyl-methylamino] phenyl [thiophene-2-sulfonylamino]-3-methylbutyric acid (TISAM) therapeutic effect on experimental autoimmune myocarditis (EAM) in Lewis rats.

METHODSTreatment protocol of oral administration of 5 mg/kg TISAM once a day for 14 days was performed on EAM Lewis rats. EAM Lewis rats were divided into 3 groups: treatment in early, middle and later stage respectively (n = 20). After experiment at the designate time point, the rats were euthanatized and hearts were harvested. Cardiac inflammatory score, fibrosis score and content, and infiltration of macrophages and T lyminflammatory score, fibrosis score and content, and infiltration of macrophages and T lymphocytes, message RNA (mRNA) expression of matrix metalloproteinase (MMP)-2 and MMP-9 and protein activity of gelatinase were determined.

RESULTSTISAM treatment in early phase was invalid (treatment started from the creation of the model), treatment in middle and later phase was effective (treatment started from 7 and 14 day after the creation of the model).

CONCLUSIONInhibitor of MMP-2 can block ventricular remodeling in middle stage in EAM Lewis rats. The mechanism maybe alleviate the inflammatory cell cardiac infiltration, decrease the mRNA expression of MMP-2 at transcript level and downregulate gelatinase activity at protein level.

Animals ; Autoimmune Diseases ; drug therapy ; Female ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase Inhibitors ; therapeutic use ; Myocarditis ; drug therapy ; Rats ; Rats, Inbred Lew ; Thiophenes ; therapeutic use

OBJECTIVETo investigate the iodine levels of urine from 1 month old breast-fed infants and the ones of milk and urine from the lactating women, and to observe the effects of different feeding methods (breast-feeding, mixed-feeding, bottle-feeding) on the iodine status of the infants during the weaning period in Beijing.

METHODSFrom March, 2001 to March, 2002, the iodine levels of urine from 97 breast-fed infants 1 month of age and the ones in milk and urine from lactating women were measured and compared. The infants followed up were divided into 3 groups (breast-fed, mixed and bottle-fed) until 6 months old. Their iodine levels of urine were measured and compared with the ones of 1 month of age.

RESULTSThe median value of urine iodine from breast-fed 1 month old infants was 183 micro g/L, suggesting that the infants with breast-fed had good iodine nutritional status. The median value of urine iodine from lactating women was 122 micro g/L, significantly lower than the value of milk iodine, 201 micro g/L (P < 0.001). which suggests that the lactating women were iodine deficient but could provide infants iodine adequately through breast feeding. Compared with 1 month af age, the urine iodine levels of 6 months old infants with breast-feeding increased (P < 0.001), the ones with bottle-feeding decreased significantly (P < 0.001) and the mixed-feeding group did not change (P > 0.05). The differences among 3 groups were significant (P < 0.005), the urine iodine levels of infants of both breast-feeding and mixed-feeding groups were higher than the ones of bottle-feeding. The breast-feeding group was the highest one among three groups.

CONCLUSIONThe breast-fed infants were nourished with iodine, but the lactating women were iodine deficient. Accompanied the decrease of the amount of breast milk, the iodine levels of infants urine decreased during the weaning period, some bottle-feeding infants were iodine deficient.

Bottle Feeding ; Breast Feeding ; Feeding Methods ; nursing ; Female ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Infant, Newborn ; Iodine ; urine ; Male