Objective: It has been found that the expression of Hedgehog signaling molecule Gli1 can inhibit the activity of estrogen signaling pathway. The present study is to observe the effect of estrogen receptor α (ERα) on the transactivation and expression of Gli1 in breast cancer cells, so as to study whether there is a cross talk between the two pathways. Methods: Using luciferase reporter gene transactivation analysis, we cotransfected MCF-7 cells with pGli-BS-luc, PcDNA3. 1-Gli1, pRL-CMV, pSG5-ERa or equimolar amounts of pSG5 vector. Then the cells were subjected to Luciferase Assays to analyze the change of Gli1 transactivation. The mRNA and protein expression of Gli1 following ectopic overexpression of ERa was also analyzed by quantitative real-time PCR and Western blotting analysis. Results= Expression of ERα induced the luciferase activity in a dose-dependent manner. ERα at 500 ng/well increased the activity of luciferase by 3.5 folds (P<0.001). The luciferase activity had no obvious changes after estrogen treatment. Overexpression of ERα increased the expression of Gli1 mRNA by 2 folds (P< 0.01), and obviously increased the expression of Gli1 protein. Conclusion: Overexpression of ERa in MCF-7 cells can greatly increase the transactivity of Gli1 and increase its expression, which indicates that there is a cross talk between the two transcription factors in breast cancer cells.

Objective To explore the association between C reactive protein (CRP)with prostatic cancer (PCa)and benign prostatic hyperplasia (BPH).Methods Retrospective analysis the 110 patients of the First Affiliated Hospital of Zhejiang University In January 2010 to August 2012 whose TPSA>4 ng/ml,postoperative pathology confirmed the diagnosis of 54 cases of prostate cancer,5 6 cases of benign prostatic hyperplasia.Detected serum CRP levels by using transmission turbidim-etry and TPSA levels by using chemiluminescence immunoassay of 54 PCa and 56 BPH patients.According to the Gleason score,PCa patients were divided into high-risk and low-risk PCa two groups,the differences among high-risk PCa,low-risk PCa and BPH groups were analyzed by nonparametric statistics analysis.Results The CRP level of high-risk PCa was 4.20~2.12 mg/L,the CRP level of low-risk PCa was 1.90~0.91 mg/L and the CRP levels in BNP patients was 1.49±0.87 mg/L,the high-risk and low-risk PCa serum CRP level obviously higher than that of patients with BPH,the differences were statistically significant (P<0.05).Conclusion Serum CRP levels of PCa patients were increased significantly,espe-cially in high-risk PCa patients.

Objective To observe the expression of resistin mRNA and protein in adipose tissues of obese rats,and to explore the correlations between resistin and obesity,insulin resistance.Methods Thirty SD rats were divided into control group(n=15) and high-fat diet group(n=15).The rats in control group recieved common forage.The obese and insulin resistance models were induced with high-fat diet in SD rats.The plasma insulin level was determined by double antibody radioinimunity analysis,and automatic biochemistry analyzer in plement was used to detect the plasma free fatty acid.After 11 weeks,glucose tolerance test was carried out to determine blood glucose levels at intervals(0,30,60,90,120 min).Resistin mRNA from fat pads tissue was extracted by RT-PCR,and then its protein was detected by Western blot.The data were analyzed statistically by SPSS 11.5 software.Results After 11 weeks,the rats′ weight in high-fat diet group increased obviously than that of control group,as well as fasting blood glucose,free fat acid homeostasis model assessment of insulin resistance(HOMAIR) and the glucose tolerance in high-fat diet group reduced greatly.The resistin mRNA and protein in white adipose tissues were significantly higher in the obese mice than those of control group(Pa

Adenocarcinoma, Mucinous ; pathology ; Breast Neoplasms ; pathology ; Carcinoma, Ductal, Breast ; pathology ; Carcinoma, Renal Cell ; pathology ; Carcinoma, Squamous Cell ; pathology ; Cystadenocarcinoma, Mucinous ; pathology ; Cystadenoma, Mucinous ; pathology ; Female ; Giant Cells ; pathology ; Humans ; Osteoclasts ; pathology ; Ovarian Neoplasms ; pathology ; Pancreatic Neoplasms ; pathology ; Thyroid Carcinoma, Anaplastic ; Thyroid Neoplasms ; pathology ; Tongue Neoplasms ; pathology ; Urologic Neoplasms ; pathology

BACKGROUND: More and more patients with periodontal disease require orthodontic treatments. Thus, the remodeling process and its mechanism of inflammatory periodontal tissues become a hot point during orthodontic tooth movement.OBJECTIVE: To observe the remodeling of inflammatory periodontal tissues during orthodontic tooth movement.METHODS: A total of 50 Sprague Dawley rats were randomly divided into the control and periodontitis groups. In the periodontitis group, rats were established periodontitis models. After that, all rats were prepared for orthodontic tooth movement models. The remodeling of periodontal tissues was observed by hematoxylin-eosin staining at 1, 3, 5, 7 and 14 days after orthodontic tooth movement.RESULTS AND CONCLUSION: The movement distance of the periodontitis group was greater than that of the control group. At 0-7 days after orthodontic force application, there was obviously bone resorption at the pressure side and the bone formation was inhibited at the tension side; at 14 days after force application, the bone resorption was diminished, associated with large numbers of multinucleated osteoclasts at the pressure sides in both groups. The findings showed that rats with periodontitis suffered more periodontal traumatism during orthodontic tooth movement, thus, treatment should be delayed until the inflammatory signs were controlled and the local inflammatory was eliminated.

OBJECTIVE To study the expressions of heat shock protein 70 and human papillomavirus16E7 protein in human laryngeal squamous cell carcinoma, and their relationship in the genesis of human laryngeal squamous cell carcinoma. METHODS The expressions of HSP70 and HPV16E7 protein were detected by the immunohistochemical method in 78 specimens with laryngeal squamous cell carcinoma, 24 specimens with vocal cord polyps and 10 specimens of normal laryngeal tissues. RESULTS In human laryngeal squamous cell carcinoma, vocal cord polyps and normal laryngeal tissues, the positive expression rates of HSP70 were 69.2 % , 8.3 % and 0 % respectively, with those of HPV16E7 protein being 43.6 % 4.2% and 0 % respectively. There was a significant difference of the expression rate of HSP70 or HPV16E7 protein between the laryngeal squamous cell carcinoma and the vocal cord polyps(P

OBJECTIVE To construct a prokaryotic expression plasmid encoding HPV16E7-HSP70 fusion gene for further study on the immunity of HPV16E7- HSP70 fusion protein against laryngeal carcinoma. METHODS HPV16E7 was PCR-amplified,digested by NheI and SacI,and ligated into pET28a. HSP70 was cloned into pGEMTeasy,then recut from the vector by SalI and NotI and ligated into pET28a-HPV16E7. PCR amplification, restrict enzyme digestion, DNA sequencing, IPTG induction and Western Blot were used to identify the recombinant plasmid. RESULTS Double digestion and PCR amplification of the recombinant plas- mid have shown that the size of the inserted fragment is as expected. Sequence analysis has demonstrated that the inserted fragment encodes for the HPV16E7- HSP70 fusion gene. IPTG induction and Western Blot have shown that the fusion protein is expressed suc- cessfully in the prokaryotic expression plasmid. CONCLUSION The recombinant prokaryotic expression plasmid pET28a-HPV16E7-HSP70 has been con- structed successfully.

Objective:To study the effect of invigorating the kidney and spleen complex prescription on deltamethrin intoxicated pregnant rats. Methods:Pregnant rats were divided into 5 groups. Except the blank group,other groups were orally fed 1/20LD50(6.93mg/kg)DM daily from day 1 to 15 when proved pregrancy. 4 Hours later,rats in model group were given physiological saline,other 3 groups were given high/ median /low dose of Zhuyun Formula. Half pregnant rats were killed on D15. The rest was kept until labour. The abortion rate,survival rate,delivery condition,the level of estradiol,progestogen were observed. Results:With respect to abortion rate,model group showed statistical signifi cance when compared with other groups. Model group also showed low progesterone level. With the treatment Zhuyun Ⅲ,it had desirable effect in decreasing abortion rate,and increasing the level of progesterone. The labour time of all groups was within the normal range. Conclusion:DM had toxicity on reproduction,it could lead to abortion which was related to endocrine function of pregnant rats. Zhuyun Ⅲ could regulate the above condition and relief DM toxicity.

Objective To investigate the mutations of CLCNKB gene in a family with classic Bartter syndrome. Methods Genetic DNA was extracted from peripheral blood leucocytes of family members.The coding exons and intron exon junctions of CLCNKB gene were amplyfied by PCR and sequenced directly.Fifty unrelated healthy subjects were selected to exclude the possibility of polymorphism. Results A heterozygous(missense)mutation(482T>G,L161R)was detected in the exon 4 of patients.The hetemzygous mutation(L161R)was found in the mother,while no mutation was found in the father of this family.L161R had not been reported and was a novel mutation when referring to literatures and human genomic database home and abroad.Conclusion A new CLCNKB gene mutation(L161R)is identified for the first time.

Vibrio parahaemolyticus has been considered as one of the most important foodborne bacterial pathogens.The loop-mediated isothermal amplification(LAMP) that amplifies DNA with high specificity and rapidity under an isothermal condition was applied for rapid detection of this pathogen for the first time.A set of four primers,two outer and two inner primers,was designed specifically to recognize the thermolabile hemolysin gene(tlh) of V.parahaemolyticus.The LAMP reaction mix was optimized.The most optimal reaction temperature and time of the LAMP assay for the tlh gene were 60℃ and 60min,respectively.Genomic DNAs from 28 bacterial strains including 14 V.parahaemolyticus strains were amplified using LAMP,and no amplicon was observed in other bacterial strains.The detection limit of this LAMP assay was around 90 fg of V.parahaemolyticus genomic DNA and 24 colonies forming units for pure cultures.In addition,this method was applied to detect artificially contaminated food samples,and the detection limit was 89 cfu/g for non-cultured artificially contaminated food samples.These results suggested that detection of V.parahaemolyticus by LAMP is an effective and low-cost procedure with high specificity and sensitivity that requires no specialized equipment.This assay is expected to become a valuable tool for rapid detection and identification of V.parahaemolyticus.