OBJECTIVE: To compare the acute toxicity of common injection and sustained-release preparation of norcantharidin for mice so as to identify the attenuation of the sustained-release preparation of norcantharidin. METHODS: The poloxamer 407 was used as a sustained-release vehicle for topical administration of norcantharidin, and the acute toxicity of mice treated with common injection and sustained-release preparation of norcantharidin was observed. The median lethal dose (LD(50)) was calculated by Bliss software. RESULTS: The symptoms of mice were similar between the two groups, but the appearance of symptoms in norcantharidin/poloxamer 407 group was 4 hours later than that in norcantharidin group. The LD(50) of norcantharidin administered through vein injection was 12.6 mg/kg. The LD(50) of norcantharidin/poloxamer 407 administered through intraperitoneal injection, intrahepatic injection and intramuscular injection were 19.9, 19.1 and 20.9 mg/kg, respectively, and the LD(50) of the common preparation were 13.0, 13.1 and 15.1 mg/kg, respectively. CONCLUSION: The norcantharidin/poloxamer 407 is less toxic than the equivalent dose of norcantharidin, mainly because norcantharidin/poloxamer 407 may release norcantharidin sustainedly, thus reducing norcantharidin concentration in blood.

ObjectiveTo investigate the inhibitory effect of lentivirusly-mediated ObR-siRNA on transplanted MCF-7 human breast cancer cells by intratumoral injection.MethodsA model of subcutaneous implanted tumor was generated through injecting MCF-7 human breast cancer cells into the nude mice.Thirty established mice with MCF-7 breast cancer cells xenograft were divided into 3 groups randomly,and mice in the experimental group were intratumorally injected with ObR-siRNA lentivirus,while the negative control group and blank control group mice were injected with the same dose of negative lentivirus and normal saline.All mice were subcutaneously injected with recombinant human leptin around the tumor site once a day.Tumor size was blindly measured every other day and the mRNA expression and protein expression levels of ObR in each group were determined.ResultsKnockdown of ObR-treated xenografted nude mice with a high leptin microenvironment was successfully established.Local injection of ObR-siRNA lentivirus significantly suppressed the established tumor growth in nude mice(P ＜ 0.01,P ＜0.01 ).Real time-PCR and Western blotting showed that the mRNA and protein expression of ObR was decreased in the ObR-siRNA lentivirus group( P ＜ 0.01,P ＜ 0.01 ).ConclusionsIntratumoral injection of recomhinant ObR-siRNA lentivirus inhibits the growth of MCF-7 cells xenografts in the nude mice,suggesting that ObR might represent a therapeutic target in the genotherapies of human breast cancer.

OBJECTIVE: To establish a multidrug-resistant cell line BEL-7402/5-FU of hepatocellular carcinoma (HCC). METHODS: BEL-7402/5-FU was induced by pulse therapy combined with continuous stepwise exposure to 5-fluorouracil in vitro. MTT assay was used to determine its multidrug resistance (MDR). Biological characteristics of the BEL-7402/5-FU cell line were observed including morphological changes, cell growth curve, population doubling time, plate cloning efficiency, adherence rate, cell cycle distribution, chromosome and tumorigenicity. Accumulation amount of adriamycin (ADM) in cytoplasm was measured by flow cytometry. The protein expression of thymidylate synthase (TS) was evaluated by immuno-cytochemical method. RESULTS: The acquired MDR cell line of BEL-7402/5-FU was established successfully. The BEL-7402/5-FU cells showed cross-resistance to ADM, vincristine (VCR), methotrexate (MTX) and oxaliplatin (OHP), whereas still sensitive to hydroxycamptothecin (HCPT). The BEL-7402/5-FU cells tended to grow in clusters in vitro. It was found that the population doubling time of BEL-7402/5-FU cells was longer than that of its parental cells. The plate cloning efficiency and the adherence rate of BEL-7402/5-FU cells at the 2nd and 3rd hour were both lower than those of the parental cells. The distributing proportion of BEL-7402/5-FU cells in G(0)/G(1) phase was less than that of the parental cells, whereas the distributing proportion of BEL-7402/5-FU cells in S phase was higher than that of the parental cells. The accumulation amount of ADM in cytoplasm of BEL-7402/5-FU cells was significantly lower while the expression level of TS protein of which was highly up-regulated as compared with those of the parental cells. CONCLUSION: Establishment of the human HCC cell line BEL-7402/5-FU might be beneficial to the studies of 5-Fluorouracil acquired MDR mechanisms and the selection of reversal modifiers.

BACKGROUND: Adequate preparation of donors and recipients prior to living-related donor renal transplantation, short warm and cold ischemia time for donor kidney, good histocompatibility of human leukocyte antigen match, and low postoperative rejection incidence provide feasibility for use of low-dose immunosuppressive agents after living-related donor renal transplantation. OBJECTIVE: To investigate the safety and effectiveness of low-dose calcineurin inhibitors (CNI), an immunosuppressive agent, in living-related donor renal transplantation. METHODS: A total of 38 recipients who underwent living-related donor renal transplantation at the Center of Renal Transplantation of the First Affiliated Hospital of Nanjing Medical University from January 2006 to June 2008 were randomized for treatment with mycophenolate mofetil (750 mg twice a day), prednisone, and either standard-dose CNI (n=18) or low-dose CNI (n=20) during 12 months post-transplantation. Ciclosporin A was given orally (starting dose, 6 and 4 mg/kg per day, respectively) in two divided doses to achieve the 12-hour whole blood concentration as measured by fluorescence polarization immunoassay. The starting dose of tacrolimus was 0.12 and 0.08 mg/kg per day respectively, and its whole blood concentration was measured by enzyme-multiplied immunoassay technique. After transplantation, patients were followed up. Renal function, pulmonary infection, liver dysfunction, and CNI nephrotoxicity at different time periods were compared between different regimens. RESULTS AND CONCLUSION: During 12 months post-transplantation, patient death occurred in one of 18 patients (5.6%) in the CNI standard-dose group and none of 20 patients (0%) in the CNI low-dose group. There was no significant difference in renal function and acute rejection between CNI standard-dose and CNI low-dose groups (P > 0.05). The incidence of liver dysfunction and CNI nephrotoxicity was significantly lower in the CNI low-dose group than in the CNI standard-dose group (P < 0.05). In addition, a low-dose CNI regimen helped recipients to lessen the economic burdens. These findings indicate that it is effective, safe and economical to use a low-dose CNI regimen in living-related donor renal transplantation.

ObjectivePolyamidoamine(PAMAM) dendrimers enhance the solubility of nicotinic acid. MethodsPAMAM dendrimers of generation 1 to 6 were prepared and the effect of pH and concentration of the dendrimers on the solubility enhancement of nicotinic acid was investigated. ResultsThe pH and concentration of the dendrimers influence the solubility enhancement of nicotinic acid. Conclusions Electrostatic interaction between the carboxyl group of the nicotinic acid and the amine groups of the dendrimers is involved.

Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Carcinoma, Large Cell ; metabolism ; pathology ; Diagnosis, Differential ; Histiocytic Sarcoma ; metabolism ; pathology ; surgery ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; Male ; Melanoma ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery

To analyze the distribution of traditional Chinese medicine constitution types in elderly patients with insomnia.

Objective To investigate the relationship between serum cholesterol levels and immunoglobin types and clinical stages in the patients with multiple myeloma (MM). Methods We retrospectively analyzed the blood lipid levels in 65 patients with MM at diagnosis, including total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein Al (apo-Al) and apolipoprotein B (apo-B), and explored relationship between lipid parameters and immunoglobulin types or clinical stages in patients with MM. Thirty healthy persons were served as controls. Results Of the 65 MM patients, 53.85% were IgG type, 63.1 % were at stage Ⅲ. The levels of TC, HDL-C, LDL-C, apo Al and apo B in the patients with MM were significantly lower than that in the controls (P <0.05), and TG in MM patients was no difference with that in the controls (P >0.05). Except one case of IgD type, the levels of TC, HDL-C, LDL-C, apo Al and apo B in Ig G and Ig A types of patients were significantly lower than that in the light chain type among other 64 cases (P <0.05), and TG levels in different immunoglobulin types was found no statistical differences. The levels of TC, HDL-C, LDL-C and apo A1 in the patients with stage Ⅲ were lower than that of stage I and controls (P <0.05), furthermore, the level of LDL in stage Ⅱwas lower than that in stage Ⅰ. Conclusion Hypocholesterolemia are seen in the patients with MM and serum cholesterol levels are related to MM staging.

Objective To investigate the effects of simvastatin (SV) on the proliferation,differentiation and apoptosis of human promyelocytic leukemia cell line NB4.Methods NB4 cells were incubated with SV at different concentration with or without all-trans retinoic acid (ATRA),and NB4 cells without any treatment were taken as normal control.Cells of different groups were collected at 24 h,48 h and 72 h after incubation for further detection.Morphological changes by Wright stain were performed.MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the surface CD11b expression levels,the early stage apoptosis ratio and cell necrosis ratio.Results Treated with 15 μ mol/L SV,10 μ mol/L SV and 5 μ mol/L SV respectively,with the NB4 cells growth,the cell inhibition rates gradually increased (F =7.15,P =0.000),as well as CD11b expression levels (F =3.41,P =0.014) and AnnexinVexpression levels (F =43.38,P =0.000).Furthermore the NB4 cells treated with 15 μ mol/L SV exhibited the most significant changes with cell inhibition rate of 0.96±0.02,CD11b expression level increased to (62.41±6.37) ％ and AnnexinV expression level increased to (87.38±2.94) ％ after 72 h incubation.Combination of 15 μmol/L SV with 0.5 μmol/L ATRA displayed obvious interaction for increasing CD11b expression levels (F =4.093,P =0.025),while no significant interaction for cell inhibition rates and Annexin V expression levels were observed.After 72 h incubation,the CD11b expression levels (89.46±9.13) ％ in NB4 cells treated with 15 μ mol/L SV in combination with 0.5 μ mol/L ATRA were significantly higher than those treated with ATRA (71.27±7.27) ％ and SV (62.41±6.37) ％ (t =2.71,P =0.054; t =4.37,P =0.017)' solely.Conclusion Simvastatin in vitro inhibits NB4 cell proliferation,promotes cell apoptosis,and synergistically induces cell differentiation with ATRA dose-dependently in vitro,which indicates that SV may have the effect of synergistic anti-promyelocytic potency with ATRA.

Objective To investigate the effect of simvastatin (SV) in combination with cytosine arabinoside (ARA-C) on the proliferation and apoptosis of K562 cells. Methods Human K562 cells were incubated with SV and cytosine arabinoside alone or in combination and K562 cells without any treatment were taken as normal control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detections. Morphological changes by Wright stain were performed. MTT method was used to assay the growth inhibition rate and cytoflowmetry was used to detect the early stage apoptosis ratio and cell necrosis ratio. Results Compared with Ara-C group and SV group, cells in the group treated with SV combined with Ara-C showed obvious karyopyknosis,apoptosis bodies formation and significant cell growth inhibition, which were positively correlated with culture time. Combination of 15 μmol/L SV and Ara-C showed the most significant cell growth inhibition with a inhibition rate of (72±1) % at 72 h of culture, as was significantly higher than that of 15 μmol/L SV group (45±2) % and 20 μmol/L Ara-C group (44±0) % (P ＜0.01),furthermore, combination of 15 μmol/L simvastatin and Ara-C showed synergistic inhibition with Q value of 1.24 and 1.19 at 24 h and 48 h in each. The apoptosis rates at early stage (AnnexinV) detected by flow cytometry in 20 μmol/L, 15 μmol/L and 10 μmol/L SV treated K562 cells were significantly higher than that in normal K562 cells (P ＜0.01), as were positively correlated with culture time and SV dose (P ＜0.05). There were no significant difference of early apoptosis rate between the 20 μmol/L SV and 15 μmol/L SV groups (P ＞0.05), yet the very two were both higher than that of 10 μmol/L SV group (P ＜0.05). There were no statistic differences of late apoptosis rate (PI) amongdifferent treated groups (P ＞0.05). Conclusion SV inhibited K562 cell proliferation and induced cell apoptosis in vitro, and combination of SV and Ara-C exhibited obvious synergistic inhibition and apoptosis, which may increase the sensitivity of K562 cell to chemotherapy. SV at 15 μmol/L may be the best concentration for K562 cells in vitro.