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 compare the effect of lymph node dissection by VATS lung cancer resection surgery and conventional lung cancer surgery.Metnods 120 patients with non-small cell lung cancer were selected and randomly divided into observation group (n =60) and control group (n =60) according to the digital table.The control group was given conventional resection of lung cancer treatment,the observation group was assisted thoracoscopic lung resection treatment.The postoperative drainage,postoperative ambulation time,postoperative hospital stay,the number of lymph node metastasis and incidence rate of postoperative complications were observed.Results The time of postoperative drainage,get out of bed and postoperative hospital stay of the observation group were all shorter than those of the control group [(2.43 ± 0.54) d vs.(5.82 ± 1.35) d,(1.23 ± 0.29) d vs.(3.87 ± 0.75) d,(2.59 ± 0.63) d vs.(6.92 ± 1.27) d;t =18.059,25.430,23.658,all P ＜ 0.05)].The lymph node dissection number of the two groups had no statistically significant difference(x2 =1.008,P ＞ 0.05).The positive rate of lymph node metastasis in the observation group was lower than that in the control group(x2 =7.033,P ＜ 0.05).The incidence rate of postoperative complication of the observation group was lower than that of the control group (x2 =6.708,P ＜ 0.05).Conclusion Video-assisted thoracoscopic lung resection for non-small cell lung cancer patients with lymph node dissection is better than conventional lung surgery,and it is worthy of studying.

Objective To observe the clinical efficacy of acupuncture-moxibustion in treating early-stage glandular cystitis. Method Forty patients diagnosed with glandular cystitis by cystoscopy and biopsy were divided into a treatment group and a control group, 20 cases in each group. The treatment group was intervened by acupuncture-moxibustion, while the control group was by intravesical perfusion chemotherapy. The clinical efficacies were compared between the two groups. Result The therapeutic efficacy of the treatment group was significantly superior to that of the control group (P<0.05), and the relapse rate confirmed by using cystoscopy and biopsy was significantly lower in the treatment group than in the control group (P<0.05). Conclusion Acupuncture-moxibustion can enhance the clinical efficacy in treating early-stage glandular cystitis.

Objective To establish a SYBR Green based real-time fluorescence quantitative PCR method for detecting human Annexin Ⅱ mRNA expression,and to detect the level of Annexin Ⅱ mRNA in human breast cancer cells MCF-7 and MDA-MB-231.Methods The specific primers were designed according to the conserved sequence of human Annexin Ⅱ gene.Total RNAs were extracted from human breast cancer cells(MCF-7,MDA-MB-231),then RNAs were transcribed reversely into cDNAs.The plasmid standards were constructed.The relative expression levels of human Annexin Ⅱ mRNA in human breast cancer cells were detected by this method.Results The square(r2 )of correlation coefficient of the standard curve in this method was 0.997,the melting curve analysis showed the single peak.The the intra-batch and inter-batch variable coefficients in the pGM-T Annexin Ⅱplasmid standard substance were 6.2%,7.8% and 9.1%,12.3% respectively.The further study indicated that AnnexinⅡ mRNA in MDA-MB-231 was higher than that in MCF-7(P<0.01).Conclusion The established SYBR Green real time fluorescence quan-titative PCR method for detecting human AnnexinⅡ is of good specificity and repeatability and can be used for quantitatively detec-ting AnnexinⅡ mRNA in breast cancer cells.

17α-hydroxylase/17,20-lyase deficiency (17OHD) is a rare cause of congenital adrenal hyperplasia.The patient predominantly presents with low-renin hypertension,hypokalemia,lack of secondary sexual development,and in women with primary amenorrhea,in male with pseudohermaphroditism.We herewith analyse the clinical features of a case of 17OHD diagnosed by gene sequencing.And the etiology,clinical manifestations,genetic features,diagnosis and treatment for 17OHD were reviewed.

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.

The construction of characteristic talent training program is crucial to strengthening the development of independent college.During the construction process,efforts should be made in the following aspects:the training aim being the inheritance of its matrix with the emphasis of application ; the curriculum system emphasizing skill training as well as improvement; the training model design making full use of resources with sufficient practice; the quality improvement highlighting regional characteristics and inheriting culture.Besides,special attention should be paid to understanding the government's relevant policies,to studying relevant theories about higher education and to grasping the generality of the education aimed at undergraduates and individuality of talents in independent college.

Objective To evaluate the performance of UW2000 automatic urine system .Methods Using UW2000 automatic u‐rine system ,to evaluate intra ,inter batch precision ,linear ,carryover ,and the accuracy of the urine visible components(RBC ,WBC);at the same time intra ,inter batch precision and accuracy of of urine dry chemistry component .Results The within‐run coefficients of variation(CV% ) of RBC in high ,middle and low‐value samples were 15 .6% ,6 .8% ,20 .3% ,WBC were 16 .4% ,10 .5% ,24 .6% , respectively .Inter‐run precision of variation(CV% ) of RBC in middle and low‐value were 12 .8% ,13 .2% ,WBC were 15 .3% , 22 .8% ,respectively .The correlation coefficient of RBC was 0 .991 and the linear range was 88-19 513/μL .The correlation coeffi‐cient of WBC was 0 .997 and the linear range was 7-8 254/μL .And carryover rate was 0 .00% for RBC and 0 .06% for WBC ;The coefficient of UW2000 and microscopy on RBC and WBC were 0 .992 ,0 .995 .Within‐run precision ,inter‐run precision and accuracy for urine dry chemical composition of high ,low controls were all within the requirements of standards .Conclusion UW2000 auto‐matic urine analyzer workstation was not only save manpower ,but also with high detection speed and save cost .Importantly ,the re‐sults are reliability .In addition ,it could meet the working requirement in the general hospital ,and has great application value .

Objective To investigate the correlation of M-type phospholipase A2 receptor (PLA2R) genetic polymorphism in two single nucleotide polymorphisms (SNPs) with idiopathic membranous nephropathy (IMN) of Chinese Han population in Northeast China.Methods A total of 327 individuals were enrolled in the study including 95 adult patients with biopsy-proved IMN (IMN group) followed up for (25.4±11.6) months and 232 healthy people identified by healthy examination in China-Japan Union Hospital of Jilin University (HC group).Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotype and allele frequency of rs35771982 and rs3828323 site in PLA2R gene.The x2 test was performed to compare the distribution difference of allelic frequency and genotype frequency of the two sites in PLA2R gene between two groups.Unconditional Logistic regression analysis was used to determine the risk factor of IMN.Results IMN and HC group were matched in male predominance and body mass index (BMI).Patients with IMN were older than the healthy controls and had higher Scr,serum total cholesterol (TC),24-hour urine protein level and lower serum albumin (Alb) level,lower estimated glomerular filtration rate (eGFR) than the healthy controls (all P ＜ 0.01).The CC genotype frequency and the C allele frequency at SNP rs35771982 site of PLA2R gene in IMN group were significantly higher than those in HC group (x2=13.658,P=0.001;x2=15.315,P=9.10× 10-5),whereas there was no distribution difference of genotype and allele frequency at rs3828323 site between two groups (x2=2.844,P =0.241; x2 =2.959,P=0.085).The CC genotype at rs35771982 site in patients with IMN was not related to,age,gender,BMI,blood pressure and several laboratory indexes such as Alb,TC,Scr,eGFR and 24-hour urine protein level (all P ＞ 0.05).Unconditional Logistic regression analysis revealed that the genotype at rs35771982,age,TC,Scr and eGFR were correlated with IMN occurrence.The CC genotype at rs35771982 was the risk factor of IMN (OR =4.408,95％CI 1.488-13.058).Conclusions The CC genotype and C allele at rs35771982 site in PLA2R may be associated with the susceptibility to IMN,whereas the correlation between gene polymorphism at rs3828323 site and IMN is not demonstrated.The CC genotype at rs35771982 is the independent risk factor of IMN in Chinese Han population in Northeast China.

Objective To observe the turnaround time(TAT) of biochemistry laboratory in a certain hospital of Chongqing city,and to improve the quality of the laboratory by shortening the TAT.Methods TAT was analyzed by analyzing the daily workload,average TAT and failure rate of outpatient clinics,outpatient emergency,inpatient clinics,and inpatient emergency subjects from 2013 to 2015.The reasons for TAT prolongation were analyzed.Results The biochemical test samples were 77 060,97 129 and 105 304 from 2013 to 2015,and the annual growth rate was 26.0％ and 8.4％ respectively.TAT of the routine outpatient department samples were (78.55nu48.47)、(69.18± 37.20)、(62.82 ±21.62)min,which decreased year by year,and the difference were statistically significant(P＜0.05),and the TAT of the outpatient emergency were (64.13 ± 31.16),(59.22 ± 23.51),(66.01±37.73)min.TAT of inpatient clinics were (92.34± 53.41),(95.03±55.73) and (122.92±78.94)min from 2013 to 2015,which increased year by year,and the difference were statistically significant(P＜0.05),and the TAT of the inpatient emergency were(65.29±36.06),(62.41±30.18),(61.48±30.12)min,which decreased year by year,and the difference were statistically significant (P＜0.05).The substandard rate of samples aforementioned were 0.04％,2.99％,0.63％ and 3.69％,respectively.Conclusion TAT increases with the samples increase,it is necessary that making sure staffs more responsible in daily work,optimizing the procedure of daily biochemistry tests,improving ability of serving for clinic and patients.