Aim To investigate the inhibitive effects of trichostatin A(TSA) on telomerase activity of HL-60 cells and expression of subunit hTERT during apoptosis in vitro and its mechanism.Methods The proliferative activity of HL-60 cells was assessed using morphology and MTT assay.Cell apoptosis was confirmed using Flow Cytometer.Telomerase activity was examined using TRAP-ELISA.The expression status of telomerase subunits was analyzed using RT-PCR.Results A time-and dose-dependent inhibition was detected in HL-60 cells treated with TSA.After 48 h TSA(300 nmol?L~(-1)) treatment,the apoptotic rate detected using cytometric assay(Annexin V/PI double staining)of HL-60 cells was 42.6%.Telomerase activity and expression level of hTERT and the key subunit of telomerase decreased at 24-hour after TSA treatment.No significant changes were observed in the expression of hTR,hTP and the other two subunits of telomerase.Conclusion TSA inhibits telomerase activity and induces apoptosis in HL-60 cells.The underlying mechanism might be related to the down regulation of hTERT transcription.

The drug-loading system of DMF (dextran - magnetic layered double hydroxide - fluorouracil) was synthesized by "co-precipitation intercalated assembly - dextran composite in situ - solvent conversion" technology. The crystal-phase characteristic and slow-release performance of DMF were investigated through X-ray diffraction (XRD), infrared spectrum (IR), transmission electron microscopy (TEM), thermogravimetry (TG) and in vitro release experiment. The targeted transshipment and slow-release effect of DMF system were evaluated by in vivo animal experiment. It was showed that the XRD of DMF matched with R-sixtetragonum type layered double hydroxide and Fd-3m cubic type ferrite. IR test demonstrated that the DMF system was a supra-molecular complex consisted of Dextran (DET), magnetic layered double hydroxide (MLDH) and fluorouracil (FU) components. The two-level supra-molecular MLDH-FU presented six-edge lozenge TEM morphology, with layered characteristics. DET on the surface of DMF was capable of protecting the layered structure of MLDH-FU, improving particle dispersion properties, and strengthening the slow-release performance of the drug delivery system. The drug release model of DMF at pH 7.35 of PBS in vitro fit to the zero-order kinetics equation C = 1.1716 x 10(-5) + 4.4626 x 10(-7) t. The drug delivery system DMF could transport drugs principally to in vivo target organs with a local effect, targeted specificity, and excellent circulation transshipment performance. The pharmacokinetic process of DMF presented multi-peak phenomenon with peak attenuation and cyclic growth. The peaks appeared at 0.25, 1, 3, 5 and 9 d separately after dosing intervention. The first peak process of DMF accorded with a pharmacokinetic equation of C(FU) = 14.34 e(-0.530t) + 36.04 e(-0.321t) + 24.18 e(-0.96t), and presented the characteristic of slow absorption and fast elimination. As for subsequent peak processes, half-life increased, bioavailability increased, and plasma clearance decreased. The highest peak value of DMF was 1/37 of original value of FU, and the relative bioavailability was 419% to original FU.

Aim To observe the effect of methotrexate (MTX) on proliferation, migration and apoptosis of cultured vascular smooth muscle cells (VSMC). Methods Rabbit thoracaortic VSMC were cultured in vitro.VSMC proliferation was evaluated by cell counting and cell cycle analysis. Monolayer cell scrape was used to observe VSMC migration. Apoptosis was observed with flow cytometry, DNA gel electrophoresis and TUNEL stain. Results MTX (25～100 nmol?L -1) inhibited VSMC proliferation in a dose-dependent manner.25 nmol?L -1 and 50 nmol?L -1 MTX increased the percentage of the S phase cells and decreased the percentage of the G 2/M phase cells (P

Objective To explore the effects of silencing H2AX gene on esophageal cancer ECA109 cell xenograft growth and sensitivity to radiotherapy in nude mice.Methods BALB/c nude mice models were established by subcutaneously inoculating differently treated ECA109 cells into nude mice.By random number table method 60 nude mice were divided into six groups,including blank group,irradiation group,negative group,negative with irradiation group,silence group and silence with irradiation group.The nude mice were irradiated with 15 Gy of 6 MV X-rays 21 d after inoculation.After irradiation 48 h,half of each mice group was terminated.The tumor specimens were processed by Western blot,RT-PCR and flow cytometry were used to detect the change in protein levels,RNA levels,and cell cycle.Results The tumor volume was (42.76 ±4.40) mm3 in silence group,significantly smaller than that of the blank group (73.18±8.80) mm3 and the negative group(71.27 ±8.40) mm3(F=67.8,P＜0.01).H2AX protein expression level in the silence group was also significantly lower (0.12 ±0.03 vs.1.12 ±0.11,1.16 ± 0.08,F =34.27,P ＜ 0.01).Relative H2AX mRNA expression level in the silence group was (0.85 ± 0.31),significantly lower than that of the blank group (1.86 ± 0.26) and the negative group (1.82 ±0.24,F =39.45,P ＜ 0.01).Co-immunoprecipitation assay showed that γ-H2AX and MDC1,53BP1 obvious interaction after irradiation,which would be weakened through silencing H2AX.The tumor volumes at different groups had significant difference after irradiation (F =13.56,P ＜ 0.01).The apoptosis rate in the silence group was significantly higher than that of the blank group and the negative group [(24.15±2.25) ％ vs.(13.26±1.54) ％,(12.78±1.47) ％ (F=54.33,P＜0.01)].G2/M phase arrested in the silence with irradiation group lower than that of the radiation group (F =10.21,P ＜ 0.01).Conclusions Silencing H2AX gene expression could inhibit the growth of esophageal cancer ECA109 cell xenograft and increase the radiosensitivity of tumor.

Objective To optimize the ethanol extraction technology ofLichong ShengsuiDecoction.Methods An L9(34) orthogonal test was used in the study. The extraction rates of calycosin glycoside, icariin, baohuosideⅠ, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, astragaloside, and ginsenoside Rd were set as indexes. The influence of ethanol concentration, extraction time, extraction temperature, and amount of ethanol on the yield of Lichong ShengsuiDecoction were detected by comprehensive scoring method.Results The optimal ethanol extraction technology forLichong ShengsuiDecoction was soaking for 2 h with ten times of 60% ethanol and then reflux extracting for two times; extraction time was 1.5 h each time at 80℃.ConclusionThe optimal extraction technology is efficient, stable and feasible, which can provides data for the further study ofLichong ShengsuiDecoction.

Objective:To explore protective effect of atorvastatin on blood vessels in early stage of atherosclerosis (AS).Methods:A total of 120 patients without AS plaques,who had >2 cardiovascular risk factors and received control cardiovascular risk factors therapy,were randomly divided into four groups:control group (did not receive atorvastatin),atorvastatin 5mg group,10mg group and 20mg group (received corresponding dose of atorvastatin). All patients were followed up for six months,changes of thromboxane B2 (TXB2),6-Keto-prostaglandin F1α (6-Keto-PGF1α),brachial-ankle pulse wave velocity (baPWV),ankle brachial index (ABI)and intima-media thickness (IMT)were observed.Results:There were no significant changes in ABI and IMT between before and after treat-ment among four groups (P >0.05 all).Compared with baseline,TXB2、baPWV levels significantly rose,6-Keto-PGF1αlevel significantly decreased after treatment in control group and 5mg group;in contrast,TXB2、baPWV lev-els significantly decreased,6-Keto-PGF1αlevel significantly rose after treatment in 10mg group and 20mg group(P <0.05~ < 0.01).After treatment six-month,compared with control group and 5mg group,the TXB2 [(148.3 ± 29.2)pg/ml,(142.3±30.6)pg/ml vs.(111.5±22.8)pg/ml,(104.9 ± 17.4)pg/ml]、baPWV[(1621.1 ± 136.1) cm/s,(1597.7±125.3)cm/s vs.(1232.9±132.3)cm/s,(1178.2±155.1)cm/s]levels significantly decreased,6-Keto-PGF1α[(104.7±66.1)pg/ml,(102.2±70.3)pg/ml vs.(132.8±48.3)pg/ml,(139.1±66.3)pg/ml]level significantly rose(P <0.05~<0.01)in 10 mg group and 20 mg group.Conclusion:Atorvastatin has protective effect on blood vessels in early stage of atherosclerosis,and 10mg atorvastatin may be the minimum effective dosage to protect blood vessels.

This article was aimed to study the acute toxicities on intragastric administration of differentRadix Aconiti Lateralis Praeparataprocessed products to Beagle dogs. A total of 16 healthy and qualified Beagle dogs were randomly divided into the blank group,Pao-Fu-Pian(PFP) group,Pao-Tian-Xiong(PTX) group andHei-Shun-Pian(HSP) group according to the body weight. The intragastric administration of 4 g crude herb per kg was given. Before medication, 1 h, 24 h, and 3, 7, 14 days after medication, the body weight, food consumption, rectal temperature, electrocardiogram, blood routine and blood biochemistry were measured. The results showed that after medication, all dogs in three experimental groups were depressed. And there were significant differences in the electrolytes of blood. Among them, the HSP group was the most obvious one. The red blood cells, blood sugar and triglycerides of dogs in the PFP group had significant difference. The lymphocytes and blood sugar had significant difference of dogs in the PTX group. However, after the medication of HSP, the lymphocytes of the dogs were decreased significantly. It was concluded that the toxicity of three processed products followed the order of HSP > PFP > PTX.

10.3969/j.issn.1000-3606.2013.06.005

The expression of resistin protein in normal human abdominal, thigh, pregnant women abdominal, non-pregnant women abdominal subcutaneous adipose tissue and placenta and the relationship between obesity, type 2 diabetes mellitus (T2DM), pregnant physiological insulin resistance (IR) and gestational diabetes mellitus (GDM) was investigated. The expression of resistin protein in normal human abdominal, thigh, pregnant women abdominal, non-pregnant women abdominal subcutaneous adipose tissue and placenta was detected by using Western blotting method. Fasting serum glucose concentration was measured by glucose oxidase assay. Serum cholesterol (CHOL), serum triglycerides (TG), serum HDL cholesterol (HDL-C) and serum LDL cholesterol (LDL-C) were determined by full automatic biochemical instrument. Fasting insulin was measured by enzyme immunoassay to calculate insulin resistance index (IRI). Height, weight, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured to calculate body mass index (BMI) and body fat percentage (BF %). Resistin protein expression in pregnant women placental tissue (67 905 +/- 8441) (arbitrary A values) was much higher than that in subcutaneous adipose tissue in pregnant women abdomen (40 718 +/- 3818, P < 0.01), non-pregnant women abdomen (38 288 +/- 2084, P < 0.01), normal human abdomen (39 421 +/- 6087, P < 0.01) and thigh (14 942 +/- 6706, P < 0.001) respectively. The resistin expression in abdominal subcutaneous adipose tissue showed no significant difference among pregnant, non-pregnant women and normal human, but much higher than that in thigh subcutaneous adipose tissue (P < 0.001). Pearson analysis revealed that resistin protein was correlated with BMI (r = 0.42), fasting insulin concentration (r = 0.38), IRI (r = 0.34), BF % (r = 0.43) and fasting glucose (r = 0.39), but not with blood pressure, CHOL, TG, HDL-C and LDL-C. It was suggested that resistin protein expression in human abdominal subcutaneous adipose tissue was much higher than that in human thigh subcutaneous adipose tissue. Resistin was closely related with central obesity, leading to IR, subsequently obesity and T2DM. Resistin protein expression in placental tissue was much higher than that in subcutaneous adipose tissue in normal human abdomen, pregnant abdomen, non-pregnant women abdomen and thigh. It was indicated that resistin protein could be secreted from human placental tissue. Resistin might be one of the factors that lead to pregnant physiological IR and GDM.

Objective To observe the differences in T-lymphocyte subsets between patients with sepsis,non-sepsis and healthy controls with different ages,and to investigate the effect of cellular immune function on readmission rate.Methods The 337 patients with sepsis,350 patients with non-sepsis and 141 healthy controls were enrolled in this prospective study from general department and intensive care unit(ICU)of emergency department in Beijing Chaoyang Hospital during May 2017 to April 2018.The effects of ages on T lymphocyte subsets and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ)and the effects of T lymphocyte subsets and APACHE Ⅱ on 30-day rehospitalization rate were comparatively studied.Results In patients with non-sepsis,the values of CD3+T,CD4+T,CD8+ T,natural killer(NK)cell count were lower in elderly group than in the non-elderlygroup(956.0±85.8)×10-6/L vs.(1363.3±46.7)×10 6/L,(647.3±59.9)×10-6/L vs.(875.8±33.4) × 10-6/L,(288.9± 31.1)× 10-6/L vs.(451.2±21.2)× 10-6/L and (14.5±0.64)％vs.(15.4±0.6)％,respectively,P＜0.01.In patients with sepsis group,the values of CD3+ T,CD4+T,NK cells count were decreased and APACHE Ⅱ were increased in the elderly group as compared with the non-elderly group.The 30-day readmission rate was higher in elderly patients than in the non-elderly patients(74 cases vs.39 cases,P ＜0.01).Compared with the non-elderly patients,the elderly patients had decreased values of CD3+ T,CD4+ T,CD8+ T,NK cells,CD4+/CD8+ cell count and an increased APACHE Ⅱ (P ＜0.05).Conclusions The cellular immune function is decreased in elderly patients,which is more obvious in elderly patients with sepsis.Age,CD3+ T,CD4+ T,CD8+T and APACHE Ⅱ could be used as an independent predictor for 30 day readmission rate.