Objective To screen the formulation for tanshinone self-emulsifying drug delivery system (SEDDS) and evaluate its stability.Methods The optimal tanshinone SEDDS formulation was established through solubility experiment,emulsion examination,fully emulsified time,droplet size determination,and pseudo-ternary diagram drawing.The content and stability were evaluated by HPLC assay under the condition of illumination,high and low temperature.Results The oil phase,surfactant and co-surfactant in the optimal tanshinone SEDDS formulation were ethyl oleate,TX10,Tween 80,and isopropyl alcohol (60∶84∶21∶35).Conclusion The acquired formulation of tanshinone SEDDS is stable in the dark and room temperature.

BACKGROUND: Whehter estradiol (E2) can change the expression of peroxisome proliferator-activated receptor gamma-2(PPAR-γ2) during differentiation of bone marrow stroma cells should be studied further.OBJECTIVE: To investigate the effects of 17β-estradiol (E2) on the gene expression of PPAR-γ2 mRNA and PPAR-γ2 protein.DESIGN: Contrast observational trial.SETTING: Laboratory Department of People's Hospital of Deyang City; Laboratory Center of Chengdu Military Command General Hospital and Chengdu Bai'ao Biol-Tech Limited Company.MATERTALS: A 3-month-old female SD rat (200±20) g used to isolate bone marrow stromal cells was obtained from Animal center of Chengdu Chinese Medicine University (Medical Experimental Animal Number 11). Dulbecco's mimimum essential medium (DMEM) was obtained from Hyctone Compnay. 1α, 25(OH)2D3, dexamethasome (DEX) and E2 were purchased from Sigma Company. Total RNA kits were obtained from Omga. One step RNA PCR kit (AMV) was obtained from Takara Shuzo Co, Ltd. Northern direct HRP labeling and detection kit was purchased from PIERCE. Western blotting luminol reagent was obtained from Santa cruz.METHODS: The experiment was carried out from April 2001 to July 2002 in the Laboratory Department of People's Hospital of Deyang City, Laboratory Center of Chengdu Military Command General Hospital and Chengdu Bai'ao Biol-Tech (0, 0.1, 10, 1 000 n) was used to interfere cell differentiation for 3 days. Cultured cells were crushed with Tris-Triton X-100 PBS; activity of alkaline phosphatase was detected with Beckman CX-7 biochemical analytical device; effect of 100 g/L formalin, stained with Weigert-hematoxylin for 10 minutes, rinsed with water, differentiated with 5 g/L hydrochloric ethanol, stained with Van Gieson, desiccated with ethanol of the fractional volume of 0.95, cleared with dimethylbenzene with RT-PCR, Northern blot and Western blot during cell differentiation.protein.proliferated within 24-72 hours. Cells shaped as triangle, multiple angles and fusiform. Three days later, volume of adherent cells was increased and colony, and 10 days later, they confluenced. Bone marrow stroma cells in many generalight red to yellow). Red plasma presented synthesis of collagen. The deeper the red was, the more the collagens were.pression of PPAR-γ2 mRNA was (4.0±0.4)%, (1.7±0.2)% and (2.8±0.2)% (t=6.1, 7.2, 11.5, P＜ 0.01), which was higher crease the expression of PPAR-γ2 protein. When concentration of E2 was 0.1, 10 and 1 000 nmol/L, expression of PPAR-γ2 protein was (2.2±0.2)%, (2.6±0.2)% and (4.1 ±0.2)%, which was higher than that in 0 nmol/L E2 group [(1.2±0.10)%, t=6.6, 8.5, 13.2, P＜0.01].CONCLUSTON: E2 can inhibit expression of alkaline phosphatase and promote differentiation of bone marrow stromal cells and expression of PPAR-γ2 mRNA and PPAR-γ2 protein.

Objective To select the strains which can produce tanshinone ⅡA like its host plant Salvia miltiorrhiza bung. Methods A total of 50 strains of endophytic fungi were isolated from healthy, living and symptomless tissues of Salvia miltiorrhiza bung, among which 29 strains were obtained from the root, 14 from the stem, 3 from the leaf, 3 from the flower and 1 from the seed. Their antimicrobial activities against nine different bacteria, including both Gram-negative and Gram-positive bacteria, were measured by Oxford plate agar diffusion bioassay. Results Our data showed that all but four strains had significant antibacterial activities on at least one indicator bacterium to some extent, and five strains (DR1, DR4, DR16, DR18 and DF2) manifested quite prominent antibacterial activities against certain pathogenic bacteria. In some degree, it might indicate that this endophytic fungus isolated from the tissues of Salvia miltiorrhiza bung has a potential value as a natural antibacterial medicine as well. Thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were carried out to test selected strains, both inside and outside of the cell to see if any strain can produce tanshinone ⅡA. The result showed that extracts from three strains, labeled as DR12 (outside cell), DR21 (inside cell) and DF3 (inside cell), had a component with the same Rf value in TLC assay as that of authentic tanshinone ⅡA. The extract from DR12 (outside cell) and DR21 (inside cell) had a peak at retention time identical to that of authentic tanshinone ⅡA in HPLC. Conclusion The fungi appear to produce the bioactive compound tanshinone ⅡA, and they could be used to produce tanshinone ⅡA by fermentation. It provides a new way to synthesize this natural medicine.

OBJECTIVETo evaluate the feasibility of detecting macrophage content on atherosclerotic plaques by optical coherence tomography (OCT) technique.

METHODSThirty New Zealand white rabbits were equally divided into 3 groups at random: Control group (fed normal rabbit chow, n = 10); lipid diet group (fed regular chow supplemented with cholesterol, n = 10) and balloon injury+ lipid diet group (balloon catheter injury of the common carotid artery after 2 weeks lipid diet, n = 10). After 12 weeks, all rabbits underwent pharmacological triggering with Chinese Russell's viper venom (CRVV, 15 mg/kg, i.p.) and histamine (0.02 mg/kg, i.v.). Common carotid arteries were detected with OCT and the Movat pentachrome stain respectively. OCT and histological examination results were compared and the correlation was analyzed.

RESULTSThe intra thickness measured by Movat pentachrome stain and by the OCT was (15.2 ± 0.9) µm and (20.2 ± 7.6) µm, the medial thickness was (434.2 ± 86.5) µm and (453.8 ± 87.2) µm, the plaque thickness was (330.2 ± 87.1) µm and (392.2 ± 134.5) µm, the fibrous cap thickness was (58.3 ± 5.6) µm and (61.2 ± 4.9) µm, respectively (all P > 0.05). The normalized standard deviation of the OCT signal (NSD) was compared with immunohistochemical detection. The OCT signal within the cap is relatively homogeneous for low macrophage density in high lipid diet group. For the raw OCT data, a correlation of r = 0.846 (P < 0.01) was found between OCT NSD and a CD68 area<10%, whereas for the base 10 logarithm OCT data, a correlation of r = 0.646 (P < 0.05) was found between OCT NSD and a CD68 area<10%. In balloon injury + high lipid diet group, the OCT signal within the cap was relatively heterogeneous for high macrophage content. For the raw OCT data, a correlation of r = 0.906 (P < 0.01) was found between OCT NSD and a CD68 area >10%, whereas for the base 10 logarithm OCT data, a correlation of r = 0.593 (P < 0.05) was found between OCT NSD and a CD68 area >10%. For the raw OCT signal NSD, a range of NSDs (7.12% to 7.35%) demonstrated 100% sensitivity and specificity (Kappa value 1.0) for differentiating caps containing >10% CD68 staining. For the base 10 logarithm OCT signal, NSD values ranging from 7.81% to 7.92% provided 70% sensitivity and 75% specificity (value 0.44) for identifying caps containing >10% CD68 staining.

CONCLUSIONSOCT is an effective tool to determine macrophage content in this model. OCT imaging can clearly visualize different types of atherosclerotic plaques and provide detailed information on plaque characteristics.

Animals ; Carotid Artery, Common ; Cholesterol ; Fibrosis ; Macrophages ; Plaque, Atherosclerotic ; Rabbits ; Sensitivity and Specificity ; Tomography, Optical Coherence

BACKGROUND:Previous studies on immunosuppression and anti-rejection after organ transplantation mainly focused on effects of T lymphocytes-mediated immune response and immunosuppressive agents on T lymphocytes. Effects of dendritic cel s were unclear. The manifestation and mechanism of immunosuppressive agent effects on dendritic cel s are not identical. OBJECTIVE:To compare the effects of different immunosuppressive agents on expression and function of costimulatory molecules of dendritic cel s, and to explore the mechanism of action of immunosuppressive agents. METHODS:20μg/L rapamycin, 0.04 mg/L mycophenolate, 10μg/L tacrolimus and 1 mg/L cyclosporine A were separately added during bone marrow cel s of C57BL/6 mice were differentiated into dendritic cel s. RESULTS AND CONCLUSION:Flow cytometry results revealed that CD40 expression in each group:rapamycin0.05). One-way mixed lymphocyte reaction results displayed dendritic cel costimulatory T cel proliferation in each group:rapamycin

Objective To investigate the correlation between NRAMP1 gene polymorphisms and susceptibility to tuberculosis ( TB) in Tibetan people in Qinghai. Methods A case-control study was con-ducted in this study, involving 99 Tibetan patients with TB and 89 healthy Tibetans. The single nucleotide polymorphisms of NRAMP1 gene at rs17235409 and rs3731865 sites were detected by using TaqMan probe method. Gene cloning and sequencing typing were performed to analyze the single nucleotide polymorphisms of NRAMP1 gene at the rs17235416 site. SPASS20. 0 software was used to statistically analyze the correla-tion between NRAMP1 gene polymorphisms and susceptibility to TB in Tibetan people. Results No signifi-cant difference in the genotype frequencies of rs3731865 and rs17235409 was found between the two groups (χ2=0. 852, P=0. 356;χ2=0. 279, P=0. 597). The genotype frequencies of TGTG/TGTG and TGTG/del+del/del at the rs17235416 site were 70. 7% ( 70/99 ) and 29. 3% ( 29/99 ) in patients with TB and 86. 5% (77/89) and 13. 5% (12/89) in healthy subjects. There were significant differences in the geno-type frequencies of TGTG/TGTG and TGTG/del+del/del between the two groups (χ2=6. 870, P=0. 009). The genotypes of TGTG/del and del/del at rs17235416 were risk factors for TB ( OR=0. 376; 95%CI:0. 178-1. 794 as compared with the TGTG/TGTG genotype in Tibetan people in Qinghai. Conclusion This study suggested that the NRAMP1 gene polymorphisms at rs3731865 and rs1723409 sites had no correlation with the susceptibility to TB in Tibetans in Qinghai. However, the NRAMP1 gene polymorphisms at rs17235416 site were correlated with the susceptibility to TB. The TGTG/del alleles at the rs17235416 site might be the risk factors for tuberculosis in Tibetans in Qinghai.

OBJECTIVETo compare the clinical features and long-term outcome of patients with midventricular obstructive hypertrophic cardiomyopathy (MVOHCM) and patients with apical hypertrophic cardiomyopathy (AHCM) in China.

METHODSThis retrospective study analyzed clinical data of 66 patients with MVOHCM and 263 patients with AHCM from a consecutive single-center cohort consisting of 2 413 patients with HCM. The clinical features, cardiovascular mortality and morbidity were compared between the two groups.

RESULTSCompared with the AHCM, patients in the MVOHCM group was younger and more likely to be symptomatic over a mean follow-up of 7 years. The proportion of MVOHCM and AHCM were 2.7% (66/2 413) and 10.9% (263/2 413) (P < 0.001), respectively, in this cohort. Cardiovascular mortality of the two groups were 13.6% (9/66) and 0.8% (2/263) (P < 0.001), and cardiovascular morbidity of the two groups were 53.0% (35/66) and 14.4% (38/263) (P < 0.001).

CONCLUSIONMVOHCM is rarer, but the clinical manifestations and long-term outcomes are worse compared with AHCM in this patient cohort.

Cardiomyopathy, Hypertrophic ; Humans ; Retrospective Studies