Objective To explore a stable and feasible method for the primary culture of breast cancer-associated fibroblast (CAF), and to offer the theoretical basis for the further studies about the role of CAF in the processing of breast cancer by analyzing the biological property of CAF. Methods CAF was primary cultured with tissue block enzymolytic method. Flow cytometry (FCM) was used to detect the cell cycle. The expression of Vimentin, α-SMA and Ki-67 were detected by cell immunochemistry method. Transwell assay was used to detect the invasion and migration capacity of CAF. Results The CAF was successfullyculturedbyconditionoptimization.ImmunohistochemicalresultsshowedthatVimentin, α-SMA and Ki-67 were highly expressed in CAF.Comparedwithfibroblastsfrombreastcancersidestissues,the proliferating ability and the invasion and migration capacities of CAF were enhanced (number of invasive cells: 79.7 ± 3.5 vs. 66.3 ± 1.5, number of migrated cells: 242.3 ± 3.1 vs. 218.3 ± 2.1, both P<0.05). Conclusion CAF cultured from breast cancer has better proliferating ability and strong invasion and migration capacities.

Objective To study the curative effect of combing montelukast with inhaled corticosteroids in the treatment of cough variant asthma .Methods According to diagnosis standard guideline for diagnosis and treatment of cough in CVA,84 cases of CVA patients were chosen and divided into the treatment group (n=42)and the control group(n=42).The control group were treated with inhaled corticosteroid Budesonide .The treatment group was given monelukast based on the treatment in control group .And the clinical effects of two groups ,recurrence and the lung function index were observed .Results The total efficiency of treatment of the control group was 73.8%,and the treatment group was 95.0%.The total effective rate of observation group was apparently higher than that of the control group(χ2 =6.46,P<0.05).The lung function of the treatment group compared to the control group was significantly increased(t=2.873,P<0.05).Conclusion Montelukast combined with Glucocorticoid in the treatment of CVA can effectively improve the cure rate ,reduce the recurrence ,which can be widely used in clinical treatment .

Objective To evaluate the effect of nebulized budesonide and oral prednisolone in treatment of idiopathic pulmonary fibrosis(IPF).Methods Forty-two patients with IPF were randomized to divide into two groups,nebulised budesonide groups and oral prednisolone groups.Symptoms,signs,pulmonary function,arterial blood gases analysis and the adverse effects were measured.ResultsThere were significant improvement of VC and PaO2 of nebulized budesonide and oral prednisolone groups,but there were no significant differences between of nebulized budesonide groups and oral prednisolone groups.The adverse effects in nebulised budesonide group were obviously less than those in oral prednisolone groups.Conclusion Nebulized budesonide was an effective method in treatment of idiopathic pulmonary fibrosis.

[Objective]To investigate the clinical application of treating soft tissue defects of the feet、ankle and lower one third of the legs with reverse saphenous neurocutaneous concomitant vessel pedicled island flap.[Method]From March 1999 to June 2003,six cases of soft tissue defects of the feet,ankle and lower one third of the legs were treated with reverse saphenous neurocutaneous concomitant vessel pedicled island flap.The size of flaps were from 12 cm?7 cm to 5 cm?3 cm,aver aging 8 cm?5 cm.[Result]Flaps of 6 cases survived all showing good quality.After 10 months to 3 years follow-up,all of the six cases were thinked very good.[Conclusion]The reversed saphenous neurocutaneous concomitant vessel pedicled island flap treating soft tissue defects of the feet,the ankle and the lower one third of legs is easy to be performed.The flap has liable blood supply and does not sacrifice the major arteries.

Objective To analyze the projects funded by National Natural Science Foundation of China(NSFC) in the field of liver diseases during 2010 to 2014.Methods The projects in the field of liver disease were statistically analyzed regarding the subordinate branches and distribution of the applicants and registered institutions.Results 1.There were 624.4 projects and 321,839,200 yuans annual funded in the field of liver diseases; 2.The percentage of the key/major projects was lower than the average level; 3.Nearly half of the projects funded focus on the liver cancer; 4.19.03％ of registered institution got 80％ of funded projects.Conclusions With the continuous funding aid from NSFC,the fundamental research in this field has progressed significantly,especially in immunology.However,the overall research capacity still need to be further improved and the research direction need to be connected more closely to the clinical requirement to meet the translational medicine demand.

Objective To explore the value of preoperative D-dimer in patients with gastric cancer in judging gastric cancer metastasis and assessing prognosis.Methods Clinicopathological data of 132 patients with gastric cancer in the First Affiliated Hospital of Anhui Medical University between Jan.2010 and Jan.2011 was analyzed retrospectively in this study.All patients were divided into two groups according to the cutoff value (1.465 mg/L)of D-dimer and the relationships between D-dimer and clinicopathological data were determined by chi-square test.Moreover,the association of preoperative D-dimer and the prognosis was analyzed by Kaplan-Meier analysis and Log-rank test.Univariate and multivaritate Cox model were used to analyze the factors which might affect the survival of the patients and significant independent factors.Results A total of 132 patients were enrolled in this study in accordance with the inclusion criteria.D-dimer was positively correlated with the depth of invasion (x2 =4.996,P ＜ 0.05),age (x2 =4.311,P ＜ 0.05) and distant metastasis of gastric cancer (x2 =16.641,P ＜0.01),but not with thc gcndcr,lymph node metastasis,tumor size,the degree of differentiation and TNM stage (P ＞ 0.05).The mean D-dimer level was (1.39 ± 0.7) mg/L in distant metastasis patients and (0.97 ±0.83) mg/L in non distant metastasis patients (P =0.023),the mean plasma D-dimer level in patients alive at the 5 years after the surgery was (0.78 ± 0.58) mg/L,which was significantly lower than the amounts determined for the deceased patients (0.75 ± 0.58) mg/L (P ＜ 0.01).The Kaplan-Meier curves showed that the patients with gastric cancer had a longer time in the low D-dimer group than in the high D-dimer group,showing a significant difference between the two groups (P ＜ 0.01).Univariate analysis showed that the overall survival rate was significantly correlated with the gender,D-dimer,tumor size,the depth of invasion,lymph node metastasis and TNM stage (P all ＜ 0.05).The D-dimer was proved to be independent risk factor for the prognosis of gastric cancer by multivariate analysis(P ＜ 0.05).Conclusion According to the analysis,D-dimer may be valuable biomarker for metastasis patients,besides,D-dimer was an independent prognostic factor for patients with gastric cancer.

ING1 family is a candidate for tumor suppressor,which has three splicing isoforms named p47ING1a,p33ING1b,and p24ING1c. Study of the effect of different isoforms of ING1 on HeLa cells proliferation and its molecular mechanism would help further identifying the functional relationship of ING1 isoforms,and finding important genes regulated by ING1. Cell growth curve and cell cycle analysis were used to observe the effect of ING1a,ING1b,and ING1c on HeLa cells growth,and the result indicated that they could all inhibit HeLa cells growth by arresting cell cycle at G0/G1 phase. PCR method was used to construct the PHD domain deletions of ING1a and ING1b. ING1a,ING1b,ING1c and the PHD domain deletions 1a?C and 1b?C were then overexpressed in HeLa cells. p16INK4a,PTEN/p27Kip1 and p53/p21Waf1 protein levels were detected by Western blot. The result showed that ING1a,ING1b,ING1c,and 1a?C except for 1b?C induced p16INK4a protein expression,in which ING1c had the most powerful effect. Luciferase assay identified that overexpression of pcDNA3.1(+)-1a?C facilitated p16INK4a transcription through enhancing p16INK4a promoter activity,while pcDNA3.1(+)-1b?C repressed the p16INK4a promoter activity . In a word,it was found for the first time that except for the p53/p21Waf1 pathway,three splicing isoforms of ING1 family could also inhibit HeLa cells proliferation though upregulation of p16INK4a and PTEN,and the PHD domain deletion of ING1a enhanced p16INK4a transcription. These findings provide new clews to further study on the mechanisms of ING1 family suppressing cancer cells growth.

OBJECTIVE To analyze the pathogens distribution and the antibiotic resistant characters.METHODS Sputum culture was performed in 267 cases who suffered from lower respiratory tract infection in emergency ward from Jan 2004 to Jun 2007.The results of 183 pathogens and drug sensitivity were analyzed.RESULTS The detected 183 strains of pathogens showed that Gram-negative bacilli accounted for 77.1%(Klebsiella pneumoniae was in the first place) and Gram-positive cocci were for 22.9%(Enterococcus faecium and Staphylococcus aureus were in the first places),the total drug resistance rate of Gram-negative bacilli to gentamicin,cefazolin,SMZ/TMP,piperacillin,ciprofloxacin and aztreonam had highly reached 60.5-70%,however to meropenem,cefepime,amikacin,imipenem and Sulperazone was 32.5-38.6%.Gram-positive cocci appeared generally higher multidrug resistant,the drug resistance rate of Gram-positive cocci to penicillins and erythromycin was all 100%,but the resistantce of Gram-positive cocci to vancomycin hadn′t been found.CONCLUSIONS Gram-negative bacilli are major pathogens in lower respiratory tract infection of emergency ward,K.pneumoniae is in the first place of Gram-negative bacilli and E.faecium and S.aureus are in the first places of Gram-positve cocci,both of them appearing highly multidrug resistant,analysis of pathogens and drug sensitivity are necessary to rational antibiotics usage,and the immunological protection should be enhanced.

Objective To investigate the prognostic factors and influencing factors of lymph node ratio (LNR) in patients with Siewert Ⅱ and Ⅲ adenocarcinoma of the esophagogastric junction (AEG).Methods The retrospective case-control study was conducted.The clinicopathological data of 444 patients with Siewert Ⅱ and Ⅲ AEG who were admitted to the First Affiliated Hospital of Anhui Medical University between January 2010 and January 2011 were collected.All the 444 patients underwent radical D2 lymph node dissection,extent of lymph node dissection was inferior mediastinum,around the esophageal hiatus and celiac lymph node.Follow-up usingtelephone interview and outpatient examination was performed to detect patients' prognosis once every 3 months within 2 years postoperatively,once every 6 months from 2 vears to 5 years postoperatively and once every 12 months after 5 years up to October 2016.Observation indicators:(1) follow-up and survival situations;(2) univariate and multivariate factors analysis affecting prognosis of patients with Siewert Ⅱ and Ⅲ[AEG;(3) univariate and multivariate factors analysis affecting LNR of patients with Siewert Ⅱ and Ⅲ AEG.The survival curve and rate were respectively drawn and calculated by the Kaplan-Meier method.The univariate analysis and multivariate analysis were respectively done using the Log-rank test and the COX regression model.Results (1) Follow-up and survival situations:all the 444 patients were followed up for 1-81 months,with a median time of 52 months.The 1-,3-,5-year overall survival rates of 444 patients were respectively 93.2％,60.3％ and 45.7％.(2) Factors analysis affecting prognosis of patients with Siewert Ⅱ and Ⅲ AEG:results of univariate analysis showed that tumor diameter,tumor differentiation,invasion depth of tumor,pN staging and staging of LNR were related factors affecting prognosis of patients with Siewert Ⅱ and Ⅲ AEG,with statistically significant differences (x2 =12.332,5.898,36.045,38.847,46.464,P＜0.05).Results of multivariate analysis showed that invasion depth of tumor and staging of LNR were independent factors affecting prognosis of patients with Siewert Ⅱ and Ⅲ AEG [RR =1.393,1.411,95％ confidence interval (CI):1.137-1.708,1.106-1.801,P＜0.05].(3) Factors affecting LNR of patients with Siewert Ⅱ and Ⅲ AEG:results of univariate analysis showed that tumor diameter,tumor differentiation and invasion depth of tumor were related factors affecting LNR of patients with Siewert Ⅱ and Ⅲ AEG,with statistically significant differences (x2=20.077,12.618,36.586,P＜0.05).Results of multivariate analysis showed that tumor diameter,tumor differentiation and invasion depth of tumor were independent factors affecting LNR of patients with Siewert Ⅱ and Ⅲ AEG (OR=0.684,0.688,0.788,95％ CI:0.485-0.965,0.505-0.936,0.687-0.903,P＜0.05).Conclusions The invasion depth of tunor and staging of LNR are independent factors affecting prognosis of patients with Siewert Ⅱ and Ⅲ AEG.Tumor diameter,tumor differentiation and invasion depth of tumor are independent factors affecting LNR of patients with Siewert Ⅱ and Ⅲ AEG.