Objective To establish relative quantification in real time reverse transcription polymerase chain reaction to measure cytokine expression.Methods TNF? and GAPDH were used as target gene and internal reference respectively. Two gene fragment were cloned, vectors were purified and quantificated. Standard curves were established using a series dilution of quantificated plasmids to measure the amplification efficiency of TNF? and GAPDH. By changing reaction conditions, the amplification efficiency of two gene were nearly 100%. Ct value of TNF? and GAPDH were measured in 14 samples stimuteneously.Results The method can detected as low as 10 3 copies with the linear range was 10 3～10 9 copies, the intra assay and interassay variation was 1% and 8% respectively. TNF? increased 8 6～9 8 fold with the average 9 2 relative to untreated control group analyzed by the 2 -??Ct methods. Conclusions The method we established has fine sensitivity and reproducibility and the data analysis was simple and reliable and can be apply to any genes.

Objective To investigate the effect of combined medication of psychological intervention and retention enema on wound healing and plasma endotoxin in patients with thrombotic hemorrhoid. Methods According to the different postoperative intervention will be January 2014-2016 year in December with hemorrhoids thrombosis in our hospital group in 90 cases as control group with normal saline retention enema,the observation group with Chinese medicine retention enema+psychological intervention;the comparative analysis of two groups of patients with various experimental data and detailed records,discusses the thrombosis of hemorrhoids after the drug retention enema combined with psychological intervention effects on wound healing and plasma endotoxin. Results The observation group(traditional Chinese medicine retention enema plus psychological intervention)clinical treatment effect is better than that of control group(saline enema)clinical curative effect,clinical symptoms of the patients were better than those in control group,plasma endotoxin level was lower than the control group,the difference between groups was statistically significant (P<0.05). Conclusion Thrombosis after hemorrhoid surgery patients choose herbal retention enema+psychological intervention clinical effect significantly ,can effectively promote wound healing ,and fully reduce plasma endotoxin.

Objective To investigate the impact factors for temperature of cornea(TOC) and analyse the relationship between TOC and evaporative dry eye. Methods Two hundred and fifty-seven patients(405 eyes) with normal lacrimal secretion received dry eye tests.Patients were divided into positive group and negative group according to the results,and were randomly subdivided into 4 groups with different environment temperature(T) and relative humidity(RH).For all eyes,TOC,body surface temperature(TBS) of forehead and center corneal thickness(CCT) were measured right after blinking.The impact factors for TOC and the differences in TOC between positive group and negative group were analysed.Results TOC was positively correlated with TBS(r=0.89),T(r=0.75) and RH(r=0.60)(P

Chordoma ; metabolism ; pathology ; Diagnosis, Differential ; Elbow ; Humans ; Immunohistochemistry ; Male ; Mucin-1 ; metabolism ; Myoepithelioma ; metabolism ; pathology ; surgery ; Soft Tissue Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; Young Adult

AIM: To construct a recombinant adenovirus vector which regulated expression of mouse transforming growth factor beta (TGF-?1). METHODS: Total RNA was extracted from Balb/c mice spleen pre-treated with Con A to clone TGF-?1. pTRE-shuttle vector was used as mediator to ligate TGF-?1 gene and the backbone of the replication-incompetent adenoviral vector. The constructed recombinant adenovirus contains tetracycline-responsive element which can regulate the expression of inserted genes. Identifying the desired recombinant adenovirus, it was packaged in HEK 293 cells. Supernatant of high titer adenovirus was collected to detect the TGF-?1 gene expression by ELISA kit. RESULTS: The constructed recombinant adenovirus had been identified by restriction endonucleases cutting, sequencing and PCR. In the supernatant of infected cells, it expressed the mice TGF-?1 in high concentration to 91.16 ng/L. CONCLUSION: The mice TGF-?1 recombinant adenovirus has been built which make a basis for further research and use in gene therapy.

Objective To study the effect of continuous renal replacement therapy(CRRT) in preventing myonephropathic metabolic syndrome(MNMS) after operation of acute arterial occlusion. Methods Twenty-four patients with acute arterial occlusion were divided randomly into 2 groups: CRRT group(n=11) and control group(n=13).The patients were treated with embolectomy or revascularization.In control group,we used conventional therapy such as anti-inflammation,expansion of blood capacity,anticoagulation,and correcting acidosis and electrolyte disorder.In CRRT group,patients were treated by continuous veno-venous hemofiltration(CVVH) with 6 h during operation and 24 h after operation. Results In control group,24 h after operation,the serum potassium,blood urea nitrogen(BUN),serum creatinine(SCr),and myoglobin(Mb) were significantly increased(P

Aim To investigate the signaling basis of the biological effects of IL-6 in U937 cells. Methods We determined the effect of IL-6 on the growth and differentiation of U937 cells and the effect of IL-6 on the activation of nuclear factors in U937 cells. Results IL-6 induced macrophage differentiation of U937 cells. Differentiated cells had strengthened ANAE (acid naphthyl acetate esterase)and NBT(nitroblue tetrazolium)-reducing activities and CD54 expression was upregulated. STAT3 could be remarkably activated by IL-6 in U937 cells and STAT3 activity was dependent on dose and time course of IL-6 treatment. However, NF-IL6,NF-κ B, AP-1 and other STAT members were not activated by IL-6. Conclusion IL-6 might induce terminal differentiation of U937 cells via JAK-STAT3 pathway.

Objective To investigate the information of outpatients with acute and chronic insomnia in the department of neurology in the general hospital.Methods In the case-control study and epidemiological investigation, the sleeping information of the outpatients in Department of Neurology of Peking University Third Hospital were collected according to classification and diagnostic criteria of mental disorder in American from October 2004 to December 2004. Multiple Logistic regression was performed to analyze the associated factors for insomnia.Results Total 925 cases were investigated and divided into acute insomnia group(149 cases), chronic insomnia group (361 cases) and control group (415 cases). After adjustment for major confounders including gender, smoking, coronary artery disease, hyperlipoidemia, education and occupation, it showed that:(1) Female (OR=2.02, P=0.001), hyperplasia of prostate (OR=1.94, P=0.036), anxiety (OR=3.29, P=0.000), depression (OR=2.48, P=0.000) and leg-moving (OR=1.78, P=0.019) increased the risk of acute insomnia significantly. (2) Female(OR=2.13, P=0.000), education (OR=2.05, P=0.005;OR=2.07, P=0.003), hyperplasia of prostate (OR=1.71, P=0.023), anxiety (OR=7.42, P=0.000) and depression (OR=4.73, P=0.000) increased the risk of chronic insomnia significantly.Conclusions Female, hyperplasia of prostate, anxiety and depression are associated with acute and chronic insomnia.

Objective This study explored the protective effect of tetramethylpyrazine on myocardial ischemia/reperfusion injury via inhibiting oxidative stress. Methods Primary cultured neonatal myocytes were applied to explore the anti-ischemia/reperfusion injury property in vitro. The survival viability of myocytes was determined by MTT; enzyme activities such as lactate dehydrogenase, creatine kinase, superoxide dismutase, malondialdehyde, and nitric oxide were analyzed with assay kits; inducible nitricoxide synthase and endothelial nitric oxide synthase expressions were determined by Westernblot. Results Tetramethylpyrazine significantly improved the beating frequencies of myocytes after oxygen-glucose deprivation/reoxygenation procedure, decreased lactate dehydrogenase, creatine kinase, and malondialdehyde levels and enhanced superoxide dismutase activity.Tetramethylpyrazine also inhibited excessive production of nitric oxide through downregulating inducible nitric oxide synthase as well as upregulating endothelial nitric oxide synthase during ischemia/reperfusion injury. Conclusion Tetramethylpyrazine could significantly improve the oxidative-stress tolerance of myocytes to keep cell membrane integrity and protect the myocardial tissue of normal physiological function via an antioxidant effect and by restoring the balance between inducible nitric oxide synthase and endothelial nitric oxide synthase, while inhibiting the generation of cytotoxic concentrations of NO.

Objective To investigate the incidence and the drug-sensitivity results of mycoplasma infection in genital tract from the gynecological outpatients,in order to provide a laboratory evidence for clinical rational use of drug.Methods The drug sensitiv-ity kit was used for isolation and culture of mycoplasma,reproductive tract secretions samples of 1 067 cases of gynecological pa-tients were collected to do mycoplasma identification and drug sensitivity test.Results In 1 067 samples,the positive rate of myco-plasma were 659 samples,including ureaplasma urealyticum(Uu)positive rate was 72.99%(481/659),mycoplasma hominis (Mh) positive rate was 3.79%(25/659),Uu and Mh positive rate accounted for 23.22%(1 53/659).The drug sensitivity results of 12 kinds of antimicrobial agents showed that single infection Uu was more sensitive to josamycin,doxycycline,minocycline,tetracy-cline,clarithromycin,azithromycin and roxithromycin.Single infection Mh was more sensitive to doxycycline,josamycin,minocycline and tetracycline.Mixed infection of Uu and Mh was more sensitive to josamycin,minocycline and doxycycline.Conclusion The main pathogen isolated from patients infected with plasma in our hospital is Uu,whether single Uu or Uu,Mh mixed infection,there are differences in the sensitivity of antimicrobial,clinicians should use drug rationally according to the conditions of patients and drug sensitivity results to reduce drug resistance rate.