OBJECTIVETo explore bone marrow-derived mesenchymal stem cells (BMSC) mediated gene directed enzyme prodrug targeting anti-tumor therapy (GDEPT).

METHODSCYP1A2 gene was cloned from human hepatocytes by RT-PCR, and the eukaryote expression vector was constructed and transferred into Raji cells and human BMSCs via liposome. The targeted anti-tumor effect of BMSC-CYP1A2 cooperated with dacarbazine (DTIC) was measured. RT-PCR and Western blot were used to detect the expression of CYP1A2. Migration assay was detected with Transwell Plates. MTT was used to evaluate the growth inhibitory effect. Cell apoptosis was determined with Annexin V-FITC/PI staining by flow cytometry(FCM).

RESULTSThe results of FCM and differentiation induction were in line with the characteristics of BMSC. The expression of CYP1A2 was confirmed by RT-PCR and Western blot. Growth inhibition of Raji-CYP1A2 cells was increased with DTIC concentration in a dose-dependent manner (IC(50) was 1.67 mmol/L). However, BMSC was less sensitive to the cytotoxic effects of DTIC (IC(50) of 9.26 mmol/L and 7.53 mmol/L for BMSC-pcDNA3.1 and BMSC-CYP1A2 cells, respectively) than Raji cells did (IC(50) of 5.62 mmol/L and 1.67 mmol/L for Raji-pcDNA3.1 and Raji-CYP1A2 cells, respectively). BMSC migrated toward Raji cells in Transwell plate. BMSC-CYP1A2 cells mediated a bystander killing effect for CYP1A2-negative Raji cells when they were co-cultured with BMSC-CYP1A2 cells.

CONCLUSIONDTIC can be catalyzed by CYP1A2 in vitro. BMSC-based enzyme prodrug system of CYP1A2 and DTIC can induce lymphoma cell apoptosis targetedly via bystander killing effect.

Apoptosis ; drug effects ; genetics ; Bone Marrow Cells ; Cells, Cultured ; Cytochrome P-450 CYP1A2 ; genetics ; Dacarbazine ; pharmacology ; Genetic Vectors ; Humans ; Lymphoma ; pathology ; Male ; Mesenchymal Stromal Cells ; Transfection

OBJECTIVETo explore the effect of different doses of 1,25-(OH)(2)VitD(3) early supplementation on airway inflammation and lung inflammatory factors in baby rats with asthma.

METHODSForty male weaned Wistar rats were divided into normal group, model group, low 1,25-(OH)(2)VitD(3) group, middle 1,25-(OH)(2)VitD(3) group, high 1,25-(OH)(2)VitD(3) group using random number table (8 rats each group). The rats in low, middle and high 1,25-(OH)(2)VitD(3) groups were given 1, 4, 10 µg/kg of 1,25-(OH)(2)VitD(3) every other day by intraperitoneal injection respectively for 25 days. Except normal group, the rats in other groups were challenged with ovalbumin to establish the asthma model. The pathologic changes of lung tissue, the total white blood cell and classified cell counts in bronchoalveolar lavage fluid (BALF) were measured. The concentrations of IL-4, IL-5 and IFN-γ in serum and BALF were measured by ELISA method.

RESULTSThe level of total white blood cell counts in BALF were (5.98 ± 1.67)×10(5)/ml, (25.34 ± 4.28)×10(5)/ml, (17.24 ± 3.3)×10(5)/ml, (9.31 ± 3.37)×10(5)/ml, (45.1 ± 15.75)×10(5)/ml, respectively (F = 33.453, P < 0.01). The percent ratio of EOS in BALF were (1.44 ± 0.78)%, (17.81 ± 6.88)%, (15.00 ± 5.70)%, (8.89 ± 3.66)%, (25.88 ± 5.57)%, respectively (F = 27.299, P < 0.01). The level of IL-4 in serum of normal, model, low, middle and high-1,25-(OH)(2)VitD(3) groups were (0.62 ± 0.54), (7.57 ± 1.04), (3.58 ± 0.56), (2.70 ± 0.78) and (5.27 ± 0.30) pg/ml, respectively (F = 116.287, P < 0.01); IL-5 in resume were (32.20 ± 4.23), (67.14 ± 18.14), (37.51 ± 0.47), (40.69 ± 2.47) and (124.60 ± 36.19) pg/ml, respectively (F = 23.902, P < 0.01); IFN-γ in serum were (79.71 ± 10.08), (49.06 ± 4.46), (59.15 ± 2.51), (59.27 ± 2.33) and (53.85 ± 1.97) pg/ml, respectively (F = 39.954, P < 0.01). Also in BLAF, the IL-4 of all groups were (0.51 ± 0.30), (102.92 ± 54.61), (8.64 ± 4.07), (3.10 ± 1.28) and (33.67 ± 8.1) pg/ml, respectively (F = 24.062, P < 0.01); the IFN-γ were (247.37 ± 189.18), (43.82 ± 13.76), (81.32 ± 17.07), (86.50 ± 14.26) and (59.89 ± 34.17) pg/ml, respectively (F = 7.157, P < 0.01); the IL-5 in BALF were (38.81 ± 0.60), (80.48 ± 17.90), (45.11 ± 1.33), (43.39 ± 1.11) and (149.60 ± 45.87) pg/ml, respectively (F = 35.978, P < 0.01). Pathologic changes in lung of asthma rat groups were obvious. The lung pathologic changes in low and middle dose groups showed a significant improvement compared to the asthma group and high dosage group showed more serious pathologic changes compared to the low and middle dose groups.

CONCLUSIONIntervention with appropriate dose of 1,25-(OH)(2)VitD(3) in the early life could improve lung pathologic changes and reduce the effect of inflammatory factors in air way of baby rat asthma model. However, overdose might play detrimental effect.

Animals ; Asthma ; metabolism ; pathology ; Bronchoalveolar Lavage Fluid ; Disease Models, Animal ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-5 ; metabolism ; Lung ; metabolism ; pathology ; Male ; Pneumonia ; metabolism ; pathology ; Rats ; Rats, Wistar ; Vitamin D ; administration & dosage ; pharmacology

A field trial was carried out to study the influence of different kinds of spring topdressing on growth, yield and quality of Ligusticum chuanxiong. The results showed that the spring topdressing had effects of improving root length, tiller numbers and plant height to some extent. At the same time the chlorophyll content and dry weight accumulation especially the dry weight of root increased significantly. It also showed that the yield increased and quality was improved significantly. The effect of different treatment with urea58.7 kg x hm(-2)(N 27 kg x hm(-2)) was the best and the treatment with N,P,K the second.
Alkaloids ; metabolism ; Coumaric Acids ; metabolism ; Fertilizers ; Ligusticum ; growth & development ; metabolism ; Seasons

OBJECTIVETo investigate whether the PD-L expression in the liver cell lines transinfected with HBV (HepG2.2.15 cells) can be up-regulated after cytokines stimulating.

METHODSTo apply the liver cell lines (HepG2 cells and HepG2.2.15 cells) as a model, the cells were stimulated with IL-4, IFN-alpha and IFN-gamma (final concentration were 10 ng/ml, stimulated for 12 hours) and RT-PCR was carried out to determine the PD-L expression before and after cytokines stimulating.

RESULTSWhether or not transinfected with HBV, IFN-alpha and IFN-gamma both can induce the liver cell lines (HepG2 cells and HepG2.2.15 cells) PD-L1 expression while IL-4 can not; IL-4, IFN-alpha, IFN-gamma all can induce the PD-L2 expression in HepG2.2.15 cells which was transinfected with HBV, only IFN-gamma can induce the PD-L2 expression in HepG2 cells which was not transinfected with HBV.

CONCLUSIONIFN-alpha, IFN-gamma both can induce the PD-L1 expression in HepG2 cells and HepG2.2.15 cells, while it is easy for cytokines to induce the PD-L2 expression in HepG2.2.15 cells which was transinfected with HBV, this may provide a potential mechanism of the molecular basis for chronic HBV infection.

Cell Line, Tumor ; Cytokines ; genetics ; metabolism ; Gene Expression ; Hepatitis B ; metabolism ; pathology ; Hepatitis B virus ; Hepatocytes ; metabolism ; virology ; Humans ; Liver ; pathology

OBJECTIVETo investigate the effect of retrovirus mediated heme oxygenase (HO)-1 gene on chronic myeloid leukemia (CML) resistance cell apoptosis induced by nilotinib (AMN107).

METHODSHigh titer viral particles of pQCXIP-EGFP-HO-1 were prepared, and K562/A02 cells stably transfected with HO-1 gene was established. The expression of HO-1 in K562/A02 cells was detected by RT-PCR. After treated with AMN107 for 24 h, HO-1 mRNA and protein expression by RT-PCR and Western blot, respectively; Cell proliferation by MTT assay; bcr-abl fusion gene by RQ-PCR, and the apoptosis and cell cycle by flow cytometry.

RESULTSRecombinant retrovirus vector was constructed successfully and K562/A02/HO-1 cells were successfully set up. The expression of HO-1 in K562/A02 cells was expressed clearly. After three groups cells treated with AMN107 for 24 h, the expression of HO-1 mRNA and protein was significantly higher in gene-transfected group than in either empty vector or no-transfected group. The difference was statistically significant (P < 0.05). The cell proliferation ofs was inhibited, but the cell viability was significantly higher in gene-transfected group than in other two groups. The difference was statistically significant(P < 0.05); After treated with 10 µmol/L AMN107 for 24 h, the CT values of bcr-abl fusion gene were (18.15 ± 0.18) in K562/A02/HO-1 group, being significantly higher than that in K562/A02/LXSN (20.32 ± 0.20) and K562/A02 (20.51 ± 0.21) group, the difference was statistically significant (P < 0.05); the apoptosis rate were (17.26 ± 0.23)%, (39.47 ± 0.17)%, and (41.84 ± 0.09)%, respectively in three groups, and were (3.74 ± 0.03)%, (5.91 ± 0.08)% in K563/A02/HO-1 untreated with drug and K562/A02 untreated with drug group. The number of G(0)/G(1) phase and S phase cells markedly decreased. The cells were arrested in G(2)/M phase. But cell cycle in gene-transfected group did not change significantly.

CONCLUSIONAMN107 inhibits proliferation of CML resistance cells and induces cell apoptosis. HO-1 gene can protect CML resistance cells to apoptosis. There was a relationship between HO-1 gene and the growth of CML resistance cells.

Apoptosis ; drug effects ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; drug effects ; Heme Oxygenase-1 ; genetics ; Humans ; K562 Cells ; Pyrimidines ; pharmacology ; Retroviridae ; genetics ; Transfection

OBJECTIVETo study the effects of soybean isoflavone on liver lipid, serum lipid, antioxidant index and hepatic lipid metabolism associated factors in nonalcoholic fatty liver rats.

METHODSThirty-six male rats (SD) were randomly divided into four groups by weight: normal control group, nonalcoholic fatty liver disease (NAFLD) model control group, low-dose isoflavone treatment group (10 mg/kg) and high-dose isoflavone group (20 mg/kg), 9 rats in each group. Normal control rats were fed with D12450B (10% fat energy), model control and isoflavone intervention rats were fed with D12492 (60% fat energy). Twelve weeks later, liver lipid, serum lipid and antioxidant index were observed. Liver sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS) and peroxisome proliferators activated receptor alpha (PPAR alpha) were detected by western blotting.

RESULTSLiver triglyceride (TG) in normal control group, NAFLD model control group, low-dose isoflavone group and high-dose isoflavone group were (8.11 ± 4.13), (57.06 ± 16.95), (31.26 ± 10.48), (31.38 ± 13.25) mmol/mg protein, respectively (F = 22.569, P < 0.01); liver free fatty acid (FFA) were (0.030 ± 0.007), (0.042 ± 0.009), (0.038 ± 0.009), (0.032 ± 0.005) µmol/mg protein, respectively (F = 4.857, P < 0.01); liver superoxide dismutase (SOD) activity were (502.29 ± 23.71), (201.83 ± 16.99), (228.93 ± 21.71), (238.08 ± 15.96) U/mg protein, respectively (F = 9.555, P < 0.01); liver malondialdehyde (MDA) were (1.29 ± 0.29), (2.85 ± 0.73), (2.07 ± 0.49), (2.03 ± 0.37) nmol/mg protein, respectively (F = 13.449, P < 0.01); SREBP-1c protein expression were 0.45 ± 0.16, 1.42 ± 0.30, 1.02 ± 0.31, 0.47 ± 0.27, respectively (F = 24.515, P < 0.01); FAS protein expression were 0.27 ± 0.08, 1.97 ± 0.47, 1.35 ± 0.30, 0.49 ± 0.12, respectively (F = 60.361, P < 0.01); PPARα protein expression were 2.03 ± 0.56, 0.41 ± 0.17, 0.81 ± 0.27, 0.66 ± 0.16, respectively (F = 37.97, P < 0.01).

CONCLUSIONSoy isoflavone can reduce the hepatic lipid deposition and increase antioxidant capacity, the mechanism may be related to inhibition of SREBP-1c and activation of PPARα expression in liver.

Animals ; Fatty Acid Synthases ; metabolism ; Fatty Liver ; metabolism ; Isoflavones ; pharmacology ; Lipid Metabolism ; Liver ; drug effects ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; PPAR alpha ; metabolism ; Rats ; Rats, Sprague-Dawley ; Soybeans ; chemistry ; Sterol Regulatory Element Binding Protein 1 ; metabolism

OBJECTIVETo explore the mechanism of fine particulate matter (PM(2.5)) induced endothelial injury and the efficacy and mechanism of ginsenoside Rg1 on the inhibition of endothelium injuries in human endothelial cells exposure to PM(2.5).

METHODSHuman umbilical vein endothelial cells (HUVECs) were stimulated with various concentrations PM(2.5) (0.1, 0.2, 0.4, 0.8 mg/ml) and PM(2.5) at concentration 0.8 mg/ml induced significant endothelial injury and was chosen for the main study in the presence or absence of Rg1 (0.04 mg/ml). After 24 h treatment, cell growth A value was detected through MTT, intracellular reactive oxygen species (ROS) level through fluorescence labeling probe method and HO-1, Nrf2 mRNA expression was detected by RT-PCR.

RESULTSThe cell A value was significantly lower while the ROS fluorescence gray value and the average optical density ratio of HO-1 were significantly higher in PM(2.5) group than in the control group (all P < 0.05). The average optical density ratio of Nrf2 was similar between PM(2.5) group and control group (P > 0.05). The A value and the average optical density ratio of HO-1 were significantly higher while the ROS fluorescence gray value was significantly lower in co-treated PM(2.5) (0.8 mg/ml) + Rg1 (0.04 mg/ml) group than in the PM(2.5) (0.8 mg/ml) group (all P < 0.05).

CONCLUSIONPM(2.5) could induce human endothelial cells injury by increasing oxidative stress which could be attenuated by ginsenoside Rg1.

Cells, Cultured ; Ginsenosides ; pharmacology ; Heme Oxygenase-1 ; metabolism ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; drug effects ; Particulate Matter ; toxicity

OBJECTIVETo explore the impact of 5-fluorouracil (5-FU) on glucose metabolism and pancreatic pathology.

METHODSTwenty Wistar rats were divided into 5-FU group(n=10, chemotherapy was administered intraperitoneally to animals at a dose of 20 mg/kg daily for continuous 5 days) and control group (n=10, sodium chloride was administered intraperitoneally to animals with the same dose at the same time ). Glucose tolerance was evaluated 2 and 7 days following 5-FU treatment by serial measurement of blood glucose before and after an oral glucose load. Plasma insulin concentration was determined by radioimmunoassay. Pancreatic pathology was examined with morphological method and the ultrastructural changes of β cells were observed by transmission electron microscope.

RESULTSFasting blood glucose level was significantly higher in the 5-FU group than that in the control group [(7.6±0.9) mmol/L vs. (4.6±0.6) mmol/L at day 2; (8.9±1.0) mmol/L vs. (4.7±0.6) mmol/L at day 7, P<0.01]. Insulin releasing test indicated that the early phase insulin response to glucose load was significantly diminished in animals treated with 5-FU at day 2. Insulin level was significantly lower in the 5-FU group than that in the control group at 30 min (P<0.01). The peak secretion time of plasma insulin in 5-FU group was at 60 min, similar to the control group; and plasma insulin level decreased more slowly. Plasma insulin level was higher in 5-FU groups than in control groups on 120 min and 180 min. At day 7, Insulin level was lower in the 5-FU group than that in the control group on 60 min, and the peak secretion time of plasma insulin was delayed to 120 min. Plasma insulin level was significantly increased in 5-FU group than that in control group on 180 min(P<0.01). No gross histopathological damage to the pancreas was observed at day 2 and 7 following administration of 5-FU. The structural changes of mitochondria were mainly the quantities of secretory granule diminished at day 7 under transmission electron microscope. Dilated rough endoplasmic reticula, swollen mitochondria, and the presence of adipose drops in lysosomes were found in few cells.

CONCLUSIONS5-FU-induced hyperglycemia appears to be mediated in part by a relatively deficient insulin secretion to glucose stimulation. A relative deficiency in insulin secretion following 5-FU treatment appears to be related to β cells function impairs with islet cell ultrastructural changes induced by 5-FU.

Animals ; Blood Glucose ; drug effects ; metabolism ; Female ; Fluorouracil ; pharmacology ; Insulin ; blood ; Male ; Pancreas ; drug effects ; pathology ; Rats ; Rats, Wistar

Objective: To investigate the changes of myo cardial contractile function during myocardial stunning in calcium overload rats and the protective effects of tetrandrine. Methods: Forty-six rats were randomized into control, myocardial ischemia, myocardial stunning, low and high dose of tetrandrine groups. Another 10 rats were used to identify the calcium overload. vitamin D3 (0.3 million Unit/kg) and nicotinic acid were adm inistered. After 16 d when calcium overload occured, left anterior descending ar tery was ligated. Twenty minutes of myocardial ischemia followed by 60 min of re perfusion was induced. The contractile function parameters were determined dynam ically. At the end of experiment, myocardial cytosolic ［Ca2+］i was deter mined in various groups. In tetrandrine groups, tetrandrine (62.2 or 93.6 μmol/ kg ) was administered by gastrogavage daily.After 16 d, the rats undergone the e xperiments mentioned above. Results: Sixteen days after vitamin D3 , nicotinic acid were given, ［Ca2+］i increased by 2.6 folds (146.8±10.8 ) vs (368.5±22.6) nmol/L, (P<0.01). Whereas, ［Ca2+］i in tetrand rine groups were (210.8±16.4) and (198.6±15.3) nmol/L, which were significantl y lower than that of calcium overload group. Twenty minutes of myocardial ische mia resulted in the decrease of dp/dtmax and Vmax in all groups with the most si gnificant in stunning and calcium overload groups. The contractile function rest ored gradually after reperfusion. At all time points, dp/dtmax and Vmax in both tetrandrine groups were higher than those in both stunning and calcium overload groups. And effect with higher dose of tetrandrine were more significant than in low dose of tetrandrine. After 60 min of reperfusion, dp/dtmax in stunning, cal cium overload, low and high dose of tetrandrine groups were 49.7%, 51.5%, 71.0% and 83.4% of that in control, respectively， and Vmax were 55.0%, 49.8%, 73.9% and 77.5% of that in control, respectively. Conclusion: T he myocardial contractile function in vitamin D3-induced calcium overload gro up is impaired. On basis of myocardiocyte calcium overload, transient ischemia l eads to myocardial stunning. At the stage of ischemia, the impaired degree of my ocardial contractile function is similar to that in stunning group, suggesting a t this stage the effect of ischemia on myocardial function is greater than that of calcium overload. Tetrandrine chronically improves the myocardial function in Vitamin D3-induced calcium overload rats.

Objective To assess the efficacy and adverse-event profile of monotherapy oradd-on therapy with topiramate in elderly patients with epilepsy. Methods A multicenter, prospective,open-label, observational study of topiramate, for either monotherapy or add-on therapy, was performedamong 119 elderly patients with epilepsy in the Outpatient Department of Neurology of 52 generalhospitals in China. After the baseline evaluation, topiramate was given at the target dose of 200 mg/dayover an 8-week titration period. In the subsequent 12-week maintenance period, the topiramate dose wasadjusted (200-300 mg) according to the clinical results. The patients were followed up for 6 months, withmonthly visits and regular physical, neurological and laboratory examinations. Results After themaintenance period, 106 patients (89.1%) showed a seizure frequency reduction by 50% or greater fromthe baseline, among whom 65 patients(93.8%)received the monotherapy and 54(83.8%) had the add-ontherapy. Topiramate monotherapy and add-on therapy resulted in a complete seizure control in over 50%of the patients with various types of seizure. In patients with disease course less than 1 year, between 1and 3 years, between 4 and 6 years, and over 6 years, topiramate monotherapy resulted in seizurereduction by 92.86%, 91.67%, 100%, and 94.44%, and the add-on therapy reduced the seizure by80.00%, 85.71%, 70.00%, and 86.36%, respectively. When combined with carbamazepine, valproatesodium, phenytoin, phenol barbital, and diazepam, the total effective rate oftopiramate was 79.41%,87.50%, 85.71%, 0%, and 80.00%, respectively. Topiramate, especially in monotherapy, caused onlymild or moderate adverse effects. Conclusion Topiramate is effective and safe for either monotherapyor add-on therapy of epilepsy in elderly patients. The types of epilepsy, disease course, or the drugs usedin the add-on therapy do no obviously affect the efficacy of topiramate for seizure control.