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.

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.

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 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.

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.

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 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 establish an HPLC method for the quantitation of nine ginsenosides (Rc ,Rb1 ,Rb2 ,Re ,Rd , Rg1 ,Rg2 ,Rg3 and Rh2 ) in Panax ginseng samples .Methods An HPLC method was developed to determine the quantities of the nine ginsenosides .The determination was performed on a Zorbax SB C18 column (4 .6 mm × 250 mm ,5μm) with an Extend-C18 guard column (4 .6 mm × 12 .5 mm ,5 μm) at 35 ℃ .The mobile phase was a multi-step acetonitrile-water gradient run at a flow rate of 1 .0 ml/min .The detection wavelength was 203 nm .Results The nine ginsenosides were baseline separated with-in 120 min .The method had good linearity ,precision ,stability and reproducibility with RSDs all less than 2 .0% .The sample recoveries were between 98 .3% to 102% .The quantity of total saponins in leaves and fibrous roots of Panax ginseng ,which were measured as 48 .9 mg/g and 23 .6 mg/g ,respectively ,were higher than those in the other plant components .The amounts of total saponins in Panax ginseng hairy roots were similar to those in taproots and fruits of Panax ginseng ,which was 7 .47 mg/g .Conclusion The established HPLC method is accurate ,simple ,rapid ,precise and reproducible and could be used for the quantitation of these nine ginsenosides in Panax ginseng samples .

Objective To compare the changes of four flavonoid glycosides in the leaves of Hippophae rhamnoides L . before and after fermentation .Methods The water extract of Hippophae rhamnoides L .leaves and its fermented tea were con-centrated and desiccated .The dry extracts were dissolved in 70％ ethanol .The chromatographic separation was performed with RP-HPLC method on an Extend-C18 column (4 .6 mm × 250 mm ,5 μm) .Acetonitrile-0 .1％ formic acid was selected as mobile phase at the flow rate of 1 .0 ml/min .The detection wavelength was 356 nm and the column temperature was 30 ℃ .Results The rutin content was high in the leaves of Hippophae rhamnoides L .After fermentation ,isoquercitrin content was increased , while the contents of rutin and narcissoside were reduced and isorhamnetin-3-O-glucoside stayed unchanged .There was a good linear relationship between the concentration and peak areas of the four compounds (r>0 .9997) .The average recoveries were between 96％-103％ .Conclusion This established method is rapid and reliable ,which can be used for the quality control of Hippophae rhamnoides L .leaves and its fermented tea .

In order to explore the treatments and prognosis of pituitary adenomas in children, the experience in the treatment of 15 children with pituitary adenomas was summed up. The clinical data of 15 children with pituitary adenomas in Tongji Hospital were retrospectively analyzed from Jan.1997 to Aug. 2004, including 9 males and 6 females with the age ranging from 9-15 years old (mean 12.8). The disease duration was 3 months to 5 years (mean 2 years). Fourteen cases were subjected to the microsurgical operation through oral tanssphenoidal approach and one case to medical treatment. All cases were followed up from 1 to 5 years by imaging studies and endocrine investigation. The results showed that 13 cases obtained total removal of the tumor and one subtotal removal of tumor. No severe complications and operation death occurred. It was concluded that the tanssphenoidal approach was the optimal treatment for children with pituitary adenomas. Regular and long-term follow-up is of great importance.