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 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 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.

BACKGROUND:It is crucial for us to find an objective and sensitive means for the assessment ofthe severitv and development of dementia,so as to make instant diagnosis and examination of therapeutic effect on patientswith vascular dementias(VD).Event related potential endogenetic eomponent P300 is proved closely correlated with the cognitive management of examinees but not with stimulus property,thereby considered as one of the most valuable and objective index for brain superior psychological activity.OBJECTIVE:To study the role and significance of events related potential P300,providing objective basis for clinical diagnosis and therapeutic assessment for patients with VD.DESIGN:Non-randomized simultaneous controlled explorative study based on patients. SETTING:Neurological department and electromyography department in aprovincial hospital and neurological department in a medical college hospital.PARTICIPANTS:Between February 2000 and June 2001,64 patients were of VD at neurological clinic and ward ofthe Second Affiliated Hospital of Chongqing Medical College,including 38 males and 26 females with theaverage age of(66.0±4.6)years(VD group).Culturallevel:19 cases re-ceiyed over senior school education,31 cases of junior school education and14 cases of illiteracy.Meanwhile 60 inpatients without VD were enrolled(non-VD group)including 35 males and 25 females withthe average age of(68.0±3.7)years.Culturallevel:senior school or above,15 cases,junior school, 29 cases and illiteracy,16 cases.METHODS:Mental sate of both VD and non-VD groups were assessed with mini-mental state examination(MMSE),and tested with P300 potential observing the variability of latency and amplitude of P300 in patients.Both methods were compared between VD group and non-VD group, moreover relationship between P300 latency and MMSE scores was also explored.MAIN OUTCOME MEASURES:Variability of P300 latency and amplitude Relationship between P300 latency and MMSE scores.RESULTS:P300 latency was found prolonged in 74% and 7% of VD group and non-VD group respectively,with obvious significance between them,meanwhile prolonged P300 latency was proved closely correlated with the MMSE scores(r=0.609 6,P＜0.05).CONCLUSION:P300 latency can be considered as an obiective predictor for the earlier diagnosis and the assessment of therapeutic effects in patients with VD.

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.

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.

Background and purpose:miR-124 is considered to be a tumor suppressor in multiple tumors, including lung cancer, prostate cancer, bladder cancer, and breast cancer. However, its function is unclear in gastric cancer. This study aimed to explore the expression of miR-124 in human gastric mucosal epithelium cells and different gastric cancer cells, as well as gastric cancer tissues and matched para-cancerous tissues. The correlations of miR-124 expression with gender, age, histological grade, T stage, TNM stage, lymph node metastasis and prognosis of gastric cancer patients were analyzed.Methods:A real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) was employed for detecting the expression of miR-124 in human gastric mucosal epithelium cells and gastric cancer cells. The expression of miR-124 in gastric cancer tissues and matched para-cancerous tissues was detected by in situ hybridization.Results:The RTFQ-PCR results indicated that the expression of miR-124 was down-regulated in MKN-74, MKN-28, MKN-45, MGC-803, SGC-7901 and AGS cells compared to GES-1 cells.In situ hybridization showed that miR-124 was strongly expressed in normal gastric mucosa. However, low expression, focal positive expression or lack of miR-124 expression were observed in gastric cancer tissues. Statistical analysis showed that miR-124 was close-ly correlated to histological stage, TNM stage and node metastasis of gastric cancer patients, but not the age, gender and tumor size. The OS and DFS of the patients with low expression of miR-124 were shorter than that of the patients with high expression of miR-124. Multivariate analysis suggested that miR-124 down-regulation was an independent prog-nostic factor for survival in patients with gastric cancer.Conclusion:miR-124 is down-regulated in gastric cancer cells and tissues. The expression of miR-124 is correlated to histological stage, TNM stage, node metastasis and prognosis.