Objective To investigate the level of serological interleukin 35 (IL-35) in breast cancer patients and its clinical significance.Methods From Jan 2015 to July 2016,serum of 55 patients with breast cancer were collected from a tertiary hospital of Nanjing.Their pathological stages were also analyzed,including 12 cases of stage Ⅰ,15 cases of stage Ⅱ,12 cases of stage Ⅲ,and 15 cases of stage Ⅳ.Meanwhile,serum from 54 healthy individuals also selected.The level of serological IL-35 was determined by ELISA,and the IL-35 level,pathological stage and tumor metastasis were analyzed by correlation a nalysis.Results The serum IL-35 in patients with breast cancer was significantly elevated than health individuals.With the increase in pathological stage,IL-35 level in breast cancer patients with stage Ⅲ and Ⅳ was significantly higher compared to patients with stage Ⅰ breast cancer (P＜0.05).Further,patients with breast cancer metastasis had increased level of IL-35,compared to breast cancer patients without tumor metastasis (P＜ 0.05).Finally,Spearman correlation analysis indicated that the serum level of IL-35 in patients with breast cancer was positlvely correlated with the pathological stages (r=0.390,P=0.004) and tumor metastasis (r=0.361,P=0.008) of the patients,respectively.Conclusion High level of IL-35 was detected in serum from breast cancer patients,and its expression was highly correlated with pathological stage and tumor metastasis.Detection of IL-35 could be applied as a valuable potential biomarker for the diagnosis of breast cancer.

Objective To investigate the clinical value of urinaryα1-acid glycoprotein (AAG)in the early diagnosis of diabetic nephropathy(DN).Methods 266 cases of diabetic patients in Drum Tower Hospital Affiliated to Medical School of Nanjing University from May 2013 to October 2014 were recruited,and patients were divided into four groups based on the urine al-bumin/creatinine (ACR)levels,as follows:non-diabetic nephropathy group (Non-DN)152 cases,low urinary albumin/cre-atinine group (LUA)49 cases,high urinary albumin/creatinine group (HUA)32 cases,and very high urinary albumin/cre-atinine group (VHUA)33 cases.63 cases of healthy subjects (NC group)and 66 cases of fracture patients (stress group) were recruited as control group.The urinary AAG and other biochemical indicators were measured for all subjects.Results The level of urinary AAG in DN group was significantly higher than that in stress group,Non-DN group and NC group (t=7.951,7.985,8.021,P <0.05),and gradually increased from LUA,HUA to VHUA group (P <0.05).The urinary AAG level was positively correlated with UA,Cr,CRP and ACR (r=0.169,0.286,0.373,0.606,P <0.01).Stepwise logistic re-gression analysis showed that the urinary AAG was the independent risk factor of DN (P <0.05).Conclusion The level of urinary AAG has good application value in the early diagnosis and monitoring progression of DN.

Objective To study the iodine nutritional status and the thyroid function of pregnant women during different periods,and provide scientific basis for iodine supplementation.Methods Totally 728 pregnant women who visited the obstetric outpatient of Nanjing Drum Tower Hospital for routine prenatal care from December 2014 to August 2016 were recruited in this study,and at the same time 182 non-pregnant women were recruited as control group.The thyroid stimulating hormone (TSH) and free thyroxine (FT4) were measured by Roche 601.The urinary iodine level was measured by SR-I-100 kit.Results The median urinary iodine of 728 pregnant women was 168.24 g/L,and the median urinary iodine of those women in the T1,T2 and T3 period were 186.31,162.65 and 148.76 g/L,respectively.The TSH at T1 period was lower than T2 and T3 period (t=3.429,3.135,P＜0.05).The FT4 at T1 period was higher than T2 and T3 period (t=5.251,5.965,P＜ 0.05).The prevalence rate of thyroid disease in normal urinary iodine group was lower than that in low urinary iodine group and high urinary iodine group (x2 =4.139,4.131,P＜0.05).Conclusion There was no iodine deficiency among those pregnant women groups,but only 34.75 ％ individuals reached the appropriate iodine nutritional level,and the ratio of iodine insufficient increased with the extension of pregnancy.The whole prevalence rate of thyroid disease in abnormal urinary iodine pregnant women was obviously higher than that in normal.It is necessary to improve the pregnant women's knowledge of iodine nutrition,moreover it is suggested that urinary iodine monitoring and thyroid function should be conducted in pregnant women.

Objective To explore the molecular mechanisms of carbapenem-resistant Klebsiella pneumoniae.Methods Carba NP confirmatory test were used to detect carbapenemases.Carbapenemase genes,ESBL genes and plasmid-mediated AmpC genes were amplified by polymerase chain reaction (PCR).The genetic correlation analysis was carried out by using multiple sequence type (MLST).Results 42 out of the 50 carbapenem-resistant Klebsiella pneumoniae strains were KPC-2-positive strains,1 strain was positive for NDM-1,the other 7 strains were not detected for any carbapenemase genes.The percentages of KPC-2-positive Klebsiella pneumoniae with the bla CTX-M,bla SHV,bla TEM,bla DHA were 21.5 ％,42.9 ％,69.1％ and 4.8％ respectively.The results of MLST showed that 37 out of 42 KPC-2 positive strains were ST11.Conclusion The production of KPC-2 is the main mechanism of Klebsiella pneumoniae resistance to carbapenem and there is an outbreak of ST11 KPC-2 Klebsiella pneumoniae in this hospital.

Objective To investigate the expression and clinical significance of zinc finger protein 217 (ZNF217)in human pancreatic cancer.Methods 43 cases with pancreatic cancer undergoing surgery in the PLA 117 Hospital and People's Hospital of Ruian City from Apr .2011 to May.2014 were enrolled in the study . The pancreatic cancer and the corresponding tumor-adjacent tissues were collected .Real-time quantitative PCR (qRT-PCR)was applied to detect ZNF217 mRNA expression in pancreatic cancer (n=43)and the corresponding tumor-adjacent normal tissues.The protein expression of ZNF217 was measured by immunohistochemistry(IHC). The relationship between the expression of ZNF 217 and clinical features was analyzed by pearson chi-square test . Results The expression level of ZNF217 mRNA and protein was significantly higher in pancreatic cancer tissues than in adjacent normal tissues(P<0.05).The high expression of ZNF217 protein was positively correlated with perineural invasion, tumor size, lymphatic metastasis and advanced TNM stage (P<0.05).Conclusions The expression of ZNF217 is significantly higher in pancreatic cancer tissues than in tumor-adjacent normal tissues , and the upregulation of ZNF 217 is associated with clinicopathological features of tumor malignance .ZNF217 may become a new marker and effective therapeutic target in early diagnosis of pancreatic cancer .

Objective To study the expression of vimentin in the tongue mucosa carcinogenesis and to explore its significance in the invasion and metastasis of tongue squamous cell carcinoma .Methods The occurrence of tongue squamous cell carcinoma in rat was induced by means of 4NQO water solution ,and 56 cases in total were collected in the cancerous process ,including normal tongue mucosa ,epithelial hyperplasia ,mild dysplasia ,moderate dysplasia ,severe dysplasia and the tongue tissue specimen of squa‐mous cell carcinoma .The immunohistochemistry was used to evaluate protein expression and real‐time fluorescent quantitative PCR method was used to obtain the expression quantity of mRNA .Results In immunohistochemistry ,with the increase of degree of rat tongue mucosa dysplasia ,the positive rate of vimentin expression increases obviously .The difference between groups was statistical‐ly significant (χ2 =10 .685 ,P<0 .05) .Lesion groups compared with normal group ,their mRNA expression differences all hold sta‐tistical significance(P<0 .05);Mild dysplasia ,moderate dysplasia ,severe dysplasia and squamous cell carcinoma groups were com‐pared with epithelial hyperplasia group .The difference between squamous cell carcinoma group and epithelial hyperplasia group was statistically significant (P<0 .05) .mRNA expression of epithelial hyperplasia ,mild dysplasia ,moderate dysplasia and severe dys‐plasia were respectively 1 .22 times ,1 .28 times ,1 .29 times and 1 .42 times of that of the normal group .Conclusion During rat tongue carcinogenesis induced by 4NQO ,the expression of vimentin was increased with the increase of the degree of pathological change ,which is closely related to the invasion of tumor and could be regarded as a predictor of tongue squamous cell carcinoma .

Objective To explore the clinical application value of the expression levels of serum hypoxia induced factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),carcino-embryonic antigen(CEA)and CA125 in predicting the recurrence or metastasis of colon cancer after radical resection.Methods Totally 123 cases of colon cancer after radical surgery in our hospital from July 2009 to December 2013 were performed the retrospective analysis.The correlation between the serum markers levels with the recurrence or metastasis was dynamically analyzed.Results (1)The HIF-1αand CEA levels after operation in 123 cases of co-lon cancer were decreased significantly compared with before operation;(2)The HIF-1α,CEA and CA125 levels in 39 cases of re-currence or metastasis were significantly increased compared with those in 84 cases of non-recurrence or non-metastasis (P <0.05);(3)in 39 cases of recurrence or metastasis,the HIF-1α and CEA levels in recurrence or metastasis were increased signifi-cantly compared with those after the end of chemotherapy (P <0.05);(4)HIF-1αwas positively correlated to VEGF(P <0.05). Conclusion The serum levels of HIF-1αand CEA in the patients with colon cancer are positively correlated with recurrence or me-tastasis,the dynamically combined detection of serum markers has an important significance for predicting and early find recurrence or metastasis after radical operation in colon cancer.

The osmotic pressure of ammonium sulfate solutions has been measured by the well-established freezing point osmometry in dilute solutions and we recently reported air humidity osmometry in a much wider range of concentration. Air humidity osmometry cross-validated the theoretical calculations of osmotic pressure based on the Pitzer model at high concentrations by two one-sided test (TOST) of equivalence with multiple testing corrections, where no other experimental method could serve as a reference for comparison. Although more strict equivalence criteria were established between the measurements of freezing point osmometry and the calculations based on the Pitzer model at low concentration, air humidity osmometry is the only currently available osmometry applicable to high concentration, serves as an economic addition to standard osmometry.

Objective To investigate the role of A2B adenosine receptor(A2BAR)in 6% HES 130/0.4-induced reduction of pulmonary capillary permeability in a rat model of sepsis.Methods Fifty male SD rats weighing 250-300 g were randomly divided into 5 groups(n = 10 each): group Ⅰ sham operation(group S);group Ⅱ sepsis(group CLP);group Ⅲ ,Ⅳ,Ⅴ low,medium,high dose HES(group H1,2,3).The animals were anesthetized with intraperitoneal pentobarbital sodium 50 mg/kg.Left carotid artery and left femoral vein were cannulated for MAP and HR monitoring and fluid and drug administration.Sepsis was induced by cecal ligation and puncture (CLP).6% HES 130/0.4 7.5,15.0 and 30.0 ml/kg were infused iv over 2 h in group H1,2,3 respectively at 4 h after CLP.The animals were sacrificed at 6 h after CLP.The lungs were isolated for determination of pulmonary capillary permeability(by iv Evans blue injection),the expression of A2BAR and the contents of cAMP,protein kinase A(PKA),TNF-α,IL-6 and IL-10 in the lung tissue.Results CLP significantly increased pulmonary capillary permeability,A2BAR expression and cAMP,IL-6 and TNF-α contents in the lung tissue in group Ⅱ as compared with group S.0.6% HES 130/0.4 significantly reduced pulmonary capillary permeability,increased A2BAR expression,cAMP,PKA and IL-10 and decreased IL-6 and TNF-αcontents in the lung tissue in group H1,2,3 as compared with group CLP.6% HES 130/0.4 decreased pulmonary capillary permeability and up-regulated A2BAR expression in a dose-dependent manner.6% HES 130/0.4 15.0 ml/kg was most effective in increasing cAMP and PKA contents in the lung and depressing inflammatory response.Conclusion 6% HES 130/0.4 decreases pulmonary capillary permeability in a rat model of sepsis by up-regulating A2BAR expression in lung tissue.

Objective To establish a simple, stable, specific and sensitive method for detection of Tyzzer ’ s or-ganism by reverse dot blotting ( RDB) .Methods Primers and specific probes were designed according to the conservative sequence of Tyzzer 16S rDNA.The forward primer was labeled with biotin .The reverse dot blotting method was established followed by PCR amplification .The specificity and sensitivity of this method were determined .Next, 41 mice and 38 rats were examined by RDB , ELISA and IFA .Results The RDB method showed a high specificity , and in the testing of the 79 laboratory animals , its limit of detection was 4.5 ng/μL.Compared the results of ELISA and IFA , its consistence with ELISA was 100%and the positive rate was 7.59%(6/79), the consistence with IFA was 92.4%(73/79), and the posi-tive rate was 0%.Conclusions An accurate, sensitive and specific method in combination with PCR and RDB in detection of Tyzzer’ s organism is established in this study .