BACKGROUND: Statins can block many intracellular signal transductive pathways and suppress the proliferation of various cells by affecting the synthesis of mevalonic acid and the translation following modification of some membrane-connecting proteins.OBJETCIVE: To investigate the influence of simvastatin on the proliferation of lung fibroblasts, the synthesis of collagen and the secretion of matrix metalproteinase-2 (MMP-2).DESIGN:Completely randomized controlled study.SETTING: At Organ Transplanting Research Institute of Fuwai Cardiovasular Diseases Hospital, Peking Union Medical College.MATERIALS: This study was carried out at the Laboratory of Cardiology of Beijing Union Medical College hospital, Peking Union Medical College from June 2004 to October 2004. Lung fibroblasts derived from neonatal SD rats were co-cultured in vitro with different dosage of simvastatin of 0, 1, 5, 10,50 μmol/L, and 50 μmol/L simvastatin + 200 μmol/L mevalonic acid.METHODS: Lung fibroblasts deriving from neonatal SD rat were co-cultured with different dosage of simvastatine in vitro. Methyl thiazolyl tetrazolium colorinetry was used to detect the cell proliferation, and cell immunohistochemical assay was used to determine the collagen synthesis and meanwhile,MMP-2 content in supernatant was examined with enzyme-linked immunosorbent assay.MAIN OUTCOME MEASURES: The proliferation of fibroblasts, the synthesis of collagen and the secretion of MMP-2 due to different dosage of simvastatin intervention and simvastatin combined with mevalonic acid.presenting the expression of type Ⅰ, Ⅲ of collagen in lung fibroblasts and the level of MMP-2 in 5, 10, 50 μmol/L simvastatin group were obviously lower than those of 0 μmol/L simvastatin group (0. 520 ± 0.010, 0. 334 ± 0.011,0.260±0.012, 0.111±0.011; 0.508±0.011, 0.324±0.014, 0.232±0.015, 0. 083 ±0. 015; 0.445 ±0. 017, 0. 305 ±0. 015, 0.216 ±0. 015,0.068±0.012; 0.561±0.013, 0.361 ±0.012, 0.289±0.012, 0.140value, the mean absorbency( A value) in lung fibroblasts and the level of MMP-2 in 50 μmol/L simvastatin + 200 μmol/L mevalonic acid group were obviously higher than that of the 50 μmol/L simvastatin group(0. 567±0.015, 0.354±0.014, 0.283±0.012, 0.138±0.011, t=4.715-10.950, P ＜ 0.01).CONCLUSION: Simvastatin could suppress the fibroblast proliferation and collagen synthesis, attenuate the secretion of MMP-2 and suppress consequently the adhesion and migration of lung fibroblasts; moreover, it has the capability of anti-cell proliferation by affecting the mevalonic acid pathway.

Objective:To study the effects of recombinant human vascular endothelial growth factor on proliferation and differentiation of rat osteoblasts. Methods: Cell culturing, light microscop, MTT assay and PNPP assay were performed to observe the effects of rhVEGF on proliferation and differentiation of the cells. Results: rhVEGF could stimulate osteoblasts proliferation and differentiation. The maximal effect on proliferation was observed at a concentration of 3.125 ng/ml on the third day. ALP activities in osteoblasts were increased most significantly at a concentration of 12.5 ng/ml. Conclusion: rhVEGF may not only induce angiogenesis to facilitate bone formation, but also stimulate this process in a direct way by inducing proliferation and differentiation of osteoblasts.

Objective:To assess the efficacy and safety of acupuncture compound anesthesia(ACA)for traditional thyroidectomy. Methods:Randomized controlled trials(RCTs)studying the use of ACA for traditional thyroidectomy were retrieved from PubMed,Excerpta Medica Database(EMBASE),Cochrane Library,Web of Science,China National Knowledge Infrastructure(CNKI),Chongqing VIP Database(CQVIP),Wanfang Academic Journal Full-text Database(Wanfang),and China Biology Medicine Disc(CBM)from inception to September 30,2021.Two investigators independently extracted data and assessed the risk of bias and quality of the studies.Anesthesia effectiveness was the primary outcome,while the secondary outcomes included various pain scales,vital signs,analgesic consumption,and adverse events.Review Manager 5.3 was used for meta-analysis.Weighted mean difference(WMD),standardized mean difference(SMD),and confidence interval(CI)were used for statistical descriptions. Results:A total of 16 papers were included,involving 1228 patients.Meta-analysis showed that anesthesia effectiveness was significantly improved after adding acupuncture as an adjunct[SMD=0.62,95%CI(0.40,0.83),P<0.0001,I2=36%].Besides,ACA can also moderate vital signs and reduce the feeling of pain[SMD=-1.61,95%CI(-2.61,-0.61),P<0.00001,I2=95%],analgesic consumption,and adverse events.Subgroup analysis of the electroacupuncture(EA)group further revealed that the effectiveness of low-frequency EA[WMD=0.43,95%CI(0.30,0.55),P<0.00001,I2=15%]and the entire operative stimulation of EA[WMD=0.55,95%CI(0.33,0.77),P<0.00001,I2=0%]was significantly better than high-frequency EA and short-time stimulation of EA during the operation.Further,no significant difference existed between conventional analgesia and acupuncture analgesia. Conclusion:ACA is beneficial to traditional thyroidectomy regarding efficacy and safety when acupuncture is applied as an adjunct.However,additional high-quality studies with larger sample sizes are needed to verify the findings.

Objective To investigate de-escalation of empiric broad-spectrum antibiotics treatment for patients in intensive care unit (ICU).Methods Data of the patients discharged from ICU in the Second Affiliated Hospital of Zhejiang University from July 1 to December 31 of 2012 and from July 1 to December 31 of 2013 were retrospectively reviewed.Patients with initial use of empirical broad-spectrum antibiotics within 3 d after ICU admission were included in the study.Clinical data including status of infection,the initial empiric antimicrobial therapy,pathogens culture and adjustment of antibiotics in 5 days were analyzed.Results A total of 841 patients were discharged from ICU during the study periods and antibiotics were used in 786 (93.5％) patients.Among 786 patients,389 (49.5％) were treated empirically with broad-spectrum antibiotics,but only 269 (69.2％) had evidences of bacterial infections.Of the 389 patients with empiric antibiotics use,de-escalation of antibiotics was applied only in 6 (1.54％) patients within 5 days after the initiation of treatment.In 269 patients with evidence of infection,specimen sampling and culture were performed in 248 (92.2％) patients within 3 days,among which 165 samples were positive,and the clinical isolates were mainly multi-drug resistant gram negative bacilli and colonized bacteria in oropharyngeal cavity.De-escalation was applied only in 4 (1.49％,4/269) patients with evidences of bacterial infections.Conclusion Broad-spectrum antibiotics as initial empiric therapy is common for patients in ICU,however de-escalation of empiric therapy is rarely applied even in patients with positive results in pathogen isolation and culture.

Objective: To explore the inhibitory effect of Shiquandabu decoction on the spontaneous len tumor of the SV40 T antigen transgenic (TG) mice, and to clarify its molecular mechanism. Methods: The SV40 T antigen TG mice were randomly divided into control group (n=39) and drug treatment group (n=25). The mice in control group were fed normally, while the mice in drug treatment group were fed with Shiquandabu decoction at the 3rd week after birth, the survival time of mice was recorded. Three mice in control group and drug treatment group were randomly chosen to collect the blood from the tail vein and the amino acid levels were measured respectively 8 and 15 weeks after Shiquandabu decoction administration. Then the mice were sacrificed and the liver tissue was collected. Gene chip hybridization was used to detect the differences in the expressions of ribosomal function related genes in liver tissue of the mice in two groups and the related signal pathway was explored. Results: The survival analysis demonstrated that the survival rate of TG mice in drug treatment was higher than that in control group (P<0. 05). Compared with control group, the serum levels of alanine, valine, leucine, isoleucine, threonine, methionine, proline, tyrosine, lysine, sarcosine, citrulline, ornithine and hydroxylysine of the mice in drug treatment group 8 weeks after administration of Shiquandabu decoction were increased (P<0. 05); and the serum levels of cystathionine, taurine, methylhistidine, anserine and ethanolamine were decreased (P<0. 05). Fifteen weeks after administration, compare with control group, the serum levels of threonine and citrulline of the mice in drug teeatment group were increased (P<0. 05), but the serum levels of other amino acids had no significant difference (P>0. 05). The canonical analysis showed that thirteen genes involved in ribosomal function from 9 083 genes in liver tissue in drug treatment group had the changes compared with control group (P<0.05). Conclusion: Shiquandabu decoction can effectively prolong the lifetime of the TG mice by improving the levels of serum amino acids and promoting the liver ribosomal protein synthesis.

Objective To establish and validate an ultra performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS)method for quantification of MRX-I,a new oxazolidinone antibacterial agent,in human plasma and urine.Methods Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C8 column using an isocratic elution.The mo-bile phase consisted of acetonitrile and water (40∶60,v/v).Quantitative analysis was conducted in the multiple reaction moni-toring mode.Linezolid was used as an internal standard.Liquid-liquid extraction with ethyl acetate was used to remove impuri-ties in the plasma and urine samples.The method was validated in terms of matrix effect,recovery,precision,accuracy and stability.Results The calibration curves were linear within the range of 0.005 00-1 .00 mg/L.The lower limit of quantification was 0.005 00 mg/L for both plasma and urine samples.Retention time was less than 1 .5 min for both MRX-I and internal standard in plasma and urine.The ma-trix effect factors of plasma and urine for MRX-I was 90.4%±8.2% and 82.7%±7.9%,respectively.The recovery of MRX-I was 112.8% ± 13.4% from plasma and 105.6% ± 13.4% from urine samples,respectively.The inter- and intra-day accuracy of MRX-I was 98.9%-105.0% and 96.5%-102.6% in plasma samples,and 92.7%-98.6% and 95.1 %-105.7% in urine samples.MRX-I was stable for 24 h at room tem-perature,48 h in automatic sampler after pretreatment,and stable after 3 freeze-thaw cycles in plasma and urine.MRX-I was also stable at-40℃for eight months in plasma and six months in urine,respectively.Conclusions The UPLC-MS/MS method established in this study shows high sensitivity and specificity for determination of MRX-I in human plasma and urine.The re-sults of validation are consistent with the requirement of bioanalytical method validation.

Objective To investigate the effect of bone marrow derived endothelial progenitor cells (EPC)transplantation on microenvironments in a murine model of chronic vein thrombosis.EPCs transplantation was evaluated whether it can up-regulate thrombus organization and recanalization associated cytokines(VEGF,ANG-1 and MCP-1). Method EPCs from immature Wister rats' bone marrow were isolated using a Ficoll density gradient centrifugation,and cultured in fibronectin-coated plate in EGM-2M Vmedium.EPCs were harvested on the 10th day,then were transplanted into chronic inferior vens cava thrombus of adult Wister rat through the femoral vein.Rats were divided into three groups:blank control group(group A,sham operation),the control group(group B,the medium injected)and the experimental group(group C,EPCs injected).The rats were sacrificed after 28 days.VEGF,ANG-1 and MCP-1 mRNA was measured by real-time quantitative PCR and protein expression change by Western blotting from IVC and thrombus tissue. Results EPCs were identificated successfullv by immunohistochemistry,immunofluorescence and function,then were transplanted into chronic inferior vena cava thrombus of adult rats.After EPCs transplantation,the VEGF,ANG-1 and MCP-1 mRNA expression in group C expression was significantly up-regulated with statistical significance(P＜0.01)compared with group A and group B in IVC and thrombus tissue by real-time PCR.There was no significant difference between group A and group B (P＞0.05).VEGF,ANG-1 and MCP-1 protein expression were similar to mRNA expression.There was significant increase in group C compared to group A and group B(P＜0.01)and no statistical significance between group A and group B(P＞0.05).Conclusion EPCs deriving from bone marrow may change the microenvimnment of chronic vein thrombus through up-regulating thrombus organization and recanalization associated cytokines(VEGF,ANG-1 and MCP-1).

Objective To construct a new replicating adenovirus vector CNHK600-p53 carrying anti-tumor gene p53 and investigate its effect on hepatocarcinoma cell line PLC/PRF5. Methods The methylthiazolyl tetrazolium assay (MTT) was used to observe the killing effect of Fluorouracil, Mitomycin and Epirubicin alone or in combination with adenovirus CNHK600-p53 on PLC/PRF5. Results When 5-Fu at concentration of 400?g/ml, the inhibition rate was (65?4. 2) % ; with MMC at 1?g/ml, the rate was (41?1.9)%; and it was (65?1.8)% when EPI at concentration of 10?g/ml. With PLC/PRF5 infected by adenovirus CNHK600-p53 ( MOI = 0. 625) , the inhibition rate was significantly increased to (89?5. 3)%,(60?2.3)% and (75?1.5)% respectively; When MOI was 0.3125, 0.625, 1.25, the inhibition rate in CNHK600-p53 group and Ad-p53 group was (27?2. 5)% , (30?3. 7)% , (61?4. 3)% and(4?2.7)%, (5?3.5)%, (16?4.5)% respectively. Conclusion For hepatocarcinoma cell line PLC/PRF5 the effect of CNHK600-p53 is stronger than Ad-p53.

Objective To investigate the effects of inhibitor of growth 5 (ING5) gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells.Methods The eukaryotic ING5-expressing plasmid and GFP-empty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment.Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability (P<0.05).Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.

A method for determination of PCDD/Fs, PCBs and PCNs in soil sample was developed by using accelerated solvent extraction (ASE)-silica gel column cleanup-basic alumina column separation coupled with GC-MS/MS.The sample was extracted by ASE with Hexane-methylene chloride (Hex-DCM, 50∶50, V/V) at 120℃.The basic alumina column was used to separate PCDD/Fs, PCBs and PCNs.The extracts were eluted with Hex-DCM (95∶5, V/V) to obtain PCBs and PCNs, followed by Hex-DCM (50∶50, V/V) to obtain PCDD/Fs.The limits of detection (LOD) were in the range of 0.04-0.25 μg/L, 0.10-0.20 μg/L and 0.01-0.05 μg/L for PCDD/Fs, PCBs, PCNs, respectively.The relative standard deviations (RSDs) of average relative response factors (RRF) were below 13%.The recoveries of 13C-labeled internal standards of the three classes of analytes were 50%-95%, 51%-103% and 49%-74%, respectively.Concentrations of ∑PCDD/Fs, ∑PCBs and ∑PCNs in soil samples were 16.1-1148 pg/g, 6.6-152.6 pg/g and 10.9-99.5 pg/g, respectively.The results were consistent with that of high resolution mass spectrometer.