Objective To establish a separation method of loganin and morroniside reference substance from Fructus Corni. Methods After extracted with hot water and precipitated with alcohol, the extract of Fructus Corni was isolated and purified by macroporous resin, silica gel column chromatography, and preparative HPLC. Loganin and morroniside were identified by UV, 1H-NMR, 13C-NMR, and MS. Results The contents of loganin and morroniside was higher than 98% by normalization method of HPLC. Conclusion The developed method is simple with lower cost, by which loganin and morroniside can be used as reference substances for the qualitative and quantitative analysis of Chinese herbal medicine.

Objective:To investigate the values of immunophenotype and the Collagen type1 alpha1/Proto-oncogene Proteins c-sis (COL1A1/PDGFB) fusion gene in the diagnosis of dermatofibrosarcoma protuberans (DFSP). Methods:IHC markers and the COL1A1/PDGFB fusion gene were detected by IHC staining and interphase fluorescence in situ hybridization (FISH) in 73 cases previously diagnosed as DFSP. A total of 85 and 10 non-DFSP cases were also included as controls for IHC staining and FISH, respectively. Results:In the 73 DFSP cases, the positive detection rates for immunohistochemical marker vimentin, CD34, CD99, S100, desmin and SMA were 100%, 91.78%, 61.64%, 0, 0, and 6.85%, correspondingly. Protein expression levels in these cases varied from the control group, and CD34 ex-pression was significantly different among the differential diagnoses. The positive detection rate for the COL1A1/PDGFB fusion gene was 86.96%(60/69), whereas the gene expression in the control group was negative. Conclusion:The COL1A1/PDGFB fusion gene is a highly specific and sensitive marker in the diagnosis of DFSP. CD34 is a suitable marker for DFSP.

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.

Objective To investigate the possibility of immune tole rance induced by bone marrow-derived mesenchymal stem cells in allogeneic organ transplantation. Methods Allogeneic bone marrow-derived mesenchymal stem cells and syngeneic bone marrow cells were cotransplanted to lethally irradiated female C57BL/6 recipient mice. FACS was used to analyze the chimerism 150 days later. Mixed lymphocyte reaction and ConA induced proliferation test were performed to evaluate proliferative activity of mice spleen cells in cell-transplanted group. Skin transplantation test was done to observe immune response of cell-transplanted group mice against organ graft from donor mice. Results About 5 97% donor T cells were detected in splenocytes of cell-transplanted group mice. MLR showed that mean SI of cell-transplanted group mice was 1.79, and that of untreated group mice was 7 28. ConA induced proliferation test showed that mean SI of cell-transplanted group mice was 31 92; and that of untreated group mice was 34 99. Mean survival time of donor-derived skin graft in cell-transplanted group mice was more than 90 days; and that in untreated group mice was 8 days. Conclusion Our results showed for the first time that induction of stable mixed hematopoietic chimerism after allogeneic bone marrow derived mesenchymal stem cells transplantation lead to stable donor-specific tolerance in allogeneic host and skin graft survival from donor mice.

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.

Objective To investigate the distribution characteristics of serum pepsinogen (PG) in healthy people and its reference interval establishment .Methods 3 753 healthy people were enrolled and divided into <45-year old ,45- <60-year old and ≥60-year old group according to their ages .Double antibody sandwich enzyme-linked immunosorbent assay was used to detect PG Ⅰ ,Ⅱ . Results Detection results of serum PG Ⅰ ,PGⅡ and PGⅠ /PGⅡ of male and female healthy people in different age group showed a skewed distribution(P<0 .05) .Serum PGⅠ and PGⅠ /PGⅡlevels of females were significantly higher than males(P<0 .01) .In the same age group ,difference of serum PGⅡ levels between males and females was not statistically significant (P>0 .05) .In the same gender ,pairwise comparison of PGⅠlevels was conducted in different age groups ,and the difference showed no statistical sig-nificance(P>0 .05) .PGⅡlevel increased with age increasing (P<0 .01) while PGⅠ /PGⅡlevel increased with age reducing (P<0 .05) .Percentile method was adopted to determine the 95% reference interval ,the bilateral reference intervals (P2 .5 - P97 .5 ) was taken for PGⅠ ,unilateral upper limit(≤ P95 ) for PGⅡ and unilateral limit (≥ P5 ) for PGⅠ /PGⅡ .Conclusion The establishment of serum PG Ⅰ ,PG Ⅱ ,PG Ⅰ /PG Ⅱ reference intervals of healthy people provides a basis for the prevention and treatment for stomach disease .

Objective To explore the CT and MR imaging features of nodular fasciitis(NF), which will in return improve the standard and quality of diagnosis. Methods CT (n = 4) and MRI (n = 5) findings of pathologically proved nodular fasciitis in 9 patients were retrospectively analyzed. Results Of 9 NF, 5 were located in the lower extremities, 2 in the abdominal wall, and the others in the acoustic duct. The resected tumor size vary from 1.1 to 9. 5 cm in the largest diameter and mean (3.7±0.3) cm. All lesions appeared as a discrete solid mass on imaging. The CT value of lesions was similar to muscle. MR imaging manifested as a hypo to isointense lesion on T_1WI and hyperintense signal intensity on T_2WI. There was homogeneous or heterogeneous moderate to significant enhancement in the lesions. Conclusion CT and MRI can provide helpful information for the clinical and differential diagnosis of NF.

Objective To explore the mechanism that gold nanorods trigger apoptosis in cancer cells.Methods Gold nanorods was synthesized by gold seed growing method, and its characterization was detected; gold nanorods on cell proliferation-toxicity were evaluated by CCK-8 Kit and apoptosis were detected by flow; mitochondrial membrane potential were tested by JC-1 and activation of Caspase 9 and Caspase 3 were detected by western blot. Results The results found that gold nanorods had nontoxic to normal cells, but highly toxic to tumor cells; and with the increasing of gold nanorods’ working time, the percentage of apoptotic cancer cells was increasing; in addition to, normal cells’ mitochondrial membrane potential did not change, but cancer cells had a significant reduction in mitochondrial membrane potential.Conclusion This study proves that gold nanorods induce apoptosis through the mitochondrial apoptosis pathway.

Objective: To investigate the clinicopathological features and immunophenotype of inflammatory myofibroblastic tumor (IMT) and their relationship with IMT diagnosis and prognosis. Methods:A total of 11 IMT cases with follow-up were analyzed morpho-logically and immunohistochemically. Results:The patients included 6 men and 5 women aged 13-66 years. The tumors were found in various anatomical sites, including lung, mediastinum, liver, intra-abdominal, and bladder. Histologically, the majority of the cases com-prised spindled fibroblastic and myofibrobalstic cells accompanied by chronic inflammatory cells in a myxoid or hyalinized stroma;the rest were individual cases of abscess formation. Prognosis mala was indicated for cases with features including atypia tumor cells with two cases demonstrating epithelioid morphology and nucleoli. Immunohistochemical study showed that vimentin, ALK, SMA, S-100, CD117, and CD34 were expressed in 91%(10/11), 55%(6/11), 100%(11/11), 27%(3/11), 18%(2/11), and 9%(1/11) of IMT, respective-ly. Ki-67 was expressed from 3%-40%respectively. CK, H-caldesmon, and DOG1 were negative in all cases. Follow-up data were avail-able for 11 patients and ranged from 4 to 22 months. Data showed that 7 patients were alive with no evidence of disease;4 patients were alive with tumor, whereas 3 showed aggressive biological behavior. Conclusion:IMTs had intermediate behavior or malignant po-tential. Most IMTs with aggressive behavior showed a minority of tumor cells with atypia, epithelioid morphology, and nucleoli. High proliferation index expression, ALK, SMA, and H-caldesmon can aid in IMT diagnosis.

Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages.Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor.Statistical tests were performed to analyze the data.Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages.Blood pressure values in the elderly group were higher than those of other groups.The morbidity of hypertension in the elderly group was higher than those of the other groups.Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age.The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups.Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension.Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages.Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.