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

Exosomes are small (30 -100 nm) membrane vesicles secreted by various cell types , they mediate cell-to-cell communication by transferring mRNAs , miRNAs, and proteins.As a hotspot in the research of oncology , exosomes have potential values in the research of development , diagnosis and treatment of cancer. This paper briefly reviews the relationship between breast cancer microenvironment , drug resistance and exosomes and its progress in breast cancer diagnosis and treatment , in order to propose new diagnostic and therapeutic strategies .

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.

Objective To study the proliferation promoting effect on osteoblast cells induced by sodium orthovanadate and whether nitric oxide (NO) was involved in this process. Methods MTT assay was employed to determine cell proliferation and its inhibition. The level of NO was measured by enzyme reduction method. Results The MTT values of MC3T3-E1 cells under the 2.5,5.0,10.0 ?mol/L of sodium orthovanadate were 0.3380?0.0045, 0.3400?0.0141, 0.3840?0.0313 respectively, which were higher than in control group (0.2540?0.0167)(P

Metallothionein is a sulfhydryl-rich,and chelating a large number of metal ions,which is widely found in biological world.It is very highlighted that MT had the same physical and chemical features from different origins.Plenty of research data indicated that MTs contain 20 free sulfhydryl genes,as well as their sulfhydryl clusters,It is a multifunctional protein in health.Many experiments demonstrate that MTs may be a novel target of original new drugs because they play a role in regulating and controlling the organ fibrosis and tumor.

Objective To investigate the effect of over-expressed Cyr61 on the expression of extracellular matrix of human renal tubular epithelial cells(HKC), and explore the role of Cyr61 in the pathogenesis and development of autosomal dominant polycystic kidney disease(ADPKD). Methods Cyr61 gene cDNA was amplified by RT-PCR. A recombinant plasmid pcDNA3.1+ Cyr61 was constructed by cloning Cyr61 gene into pcDNA3.1. HKC cells were transfected with wild-type Cyr61 plasmid vector (pcDNA3.1+Cyr61). Then the fusion of Cyr61 gene and expression of its protein were detected by RT-PCR and Western blot. The mRNA expression of Cyr61, collagen Ⅰ, collagen Ⅳ, and laminin were determined in four groups (the untransfected cells, pcDNA3.1 transfected cells, pcDNA3.1+Cyr61 -transfected cells and cyst-lining epithelial cells) by fluorescence quantum PCR. Results The amount of Cyr61 protein in Cyr61-transfected cells and cyst-lining epithelial cells were much greater than the control. The mRNA expression of Cyr61, collagen Ⅰ, collagen Ⅳ and laminin in Cyr61-transfected cells were all amplified significantly, and the level of collagen Ⅳwas much higher than collagen I(P

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.

The growth-propagation and decolorization-degradation systems for the model dy e Biebrich Scarlet by Trametes hirsuta were established preliminar ily. It was showed that 30℃ and the static culture were better; the effects of compo nents in culture media on the efficiencies of decolorization and degradation wer e not significant; considering the convenience of observing the changes of dye c o lors and shortenig the culture period, the potato liquid medium had the advantag e of others as a preferable medium for reaction systems of Trametes hirsuta . The decolorization and degradation of all dyes Biebrich Scarlet and Direct Deep Blu e L-3RB, Reactive Blue X-BR, Basic Violet 5BN, and Methylene Blue by Tramete s hirsuta were better.