Objective To evaluate differences of serum bone gamma-carboxyglutamic-acid-containing protein (BGP)levels be-tween different gender in the middle-aged and elder population and the correlation between serum BGP and osteoporosis,as well as the correlation between BGP and other bone metabolic markers.Methods The study population consisted of 270 health care middle-aged and elder people,who were excluded malignancy and chronic diseases,during 2011 January to Au-gust.Of all the Recipients,101 cases were male,aged 50 to 89 years,with a median age of 68 years old,female 169 cases, aged 50 to 89 years,with a median age of 64 years.Bone density were measured by absorptiometry and was evaluated by the T index values.Serum BGP,25-hydroxyvitamin D,calcium and phosphorus were measured by different assays systems.The different level of BGP between genders was also analyzed by Mann-Whitney test.Correlation between BGP and serum calci-um,phosphorus,25-hydroxyvitamin D,osteoporosis risk index were analyzed Spearman rank correlation analysis.Results T value,BGP,25-hydroxy vitamin D,calcium and phosphorus levels range of 270 cases were-3.5~-0.7 (median-1.6 ng/ml),3.59~264.90 ng/ml (median 12.84 ng/ml),4.0~34.0 ng/ml (media 10.5 ng/ml),1.79~2.69 mmol/L (median 2.36 ng/ml),0.43~2.89 mmol/L (median 1.12 ng/ml),respectively.BGP levels in the female groups were significantly higher than the male groups.Serum concentration of BGP was positively correlated with serum phosphorus,but the serum BGP with calcium,25-hydroxy vitamin D,age and osteoporosis risk indices were not correlated.Conclusion In the elder groups,female BGP levels were significantly higher than male,the gender factor should be considered in the clinical applica-tion of BGP.Since BGP and osteoporosis risk index T had positive correlation,those two tests can be combined to evaluate osteoporosis.

Objective To study the role of D‐lactate gradient across the lung in the rapid diagnosis of pneumonia and evalua‐tion of therapeutic efficacy .Methods Patients were divided into pneumonia group (n=46) and non‐pneumonia group (n=28) in ICU .D‐lactate gradient across the lung were calculated by the difference between arterial and mixed‐venous D‐lactate concentrations before the treatment ,after 3 and 7 days of treatment .Serum procalcitonin (PCT) ,Oxygenation index ,the lung injury score (LIS) and clinical pulmonary infection score(CPIS) were recorded at the same time .Results The mean D‐lactate gradient across the lung in pneumonia group was significantly higher than that in non‐pneumonia group[(163 .84 ± 10 .72) ng/mL vs .(30 .33 ± 7 .25) ng/mL ,P<0 .01) ]before treatment .Using a cut‐off value of 106 .11 ng/mL ,D‐lactate gradient across the lung′s sensitivity for di‐agnosis pneumonia was 90 .7% and its′specificity was 75 .5% .D‐lactate gradient across the lung correlated with LIS (r= 0 .554 , P<0 .01) and CPIS(r=0 .543 ,P<0 .01) .Conclusion D‐lactate gradient across the lung correlates with lung injury and pulmonary infection positively and may be a potential biomarker for rapid diagnosis of pneumonia .

Objective To construct the recombinant X-linked inhibitor of apoptosis protein(XIAP) gene 3′untranslational region (3′UTR)-luciferase reporter vector ,and analyze the microRNA(miRNA) which possibly regulate the expression of XIAP gene . Methods Polymerase chain reaction (PCR) was employed to amplify X IA P-3′UTR sequences from human cDNA ,in which luciferase reporter vector pGL3-Ctrl was inserted ,and the recombinant vector pGL3-Ctrl/XIAP was gained .Target Scan 6 .2 soft-ware was adopted to predict the miRNA which possibly combined with the X IA P-3′UTR .pGL3-Ctrl/XIAP recombinant plasmids and the miRNA were co-transfected into A549 cells ,and the X IA P-3′UTR-luciferase activity was measured .Results Confirmed by digestion and DNA sequencing ,the X IA P-3′UTR-luciferase reporter recombinant was successfully constructed .Prediction of miRNA target sites indicated that X IA P gene may be the target of miR-200b ,miR-200c and miR-429 .Compared with miRNA mim-ic ctrl group ,miR-200b ,miR-200c and miR-429 significantly reduced the luciferase activity of pGL 3-Ctrl/XIAP with statistically significant difference(P<0 .05) .Conclusion X IA P-3′UTR-luciferase reporter vector is successfully constructed .miR-200b ,miR-200c and miR-429 can obviously decrease the luciferase activity .

Objective To investigate the expression of livin of kidney tissues in rats with acute kidney injury (AKI) induced by endotoxin ,and the role of livin in the cell apoptosis of AKI .Methods The rat models with AKI were induced by endotoxin .The de-gree of kidney injury was observed by hematoxylin and eosin stain ,measuring the levels of the serum creatinine and urea nitrogen . The expression of livin and caspase-3 in kidney tissue at different time points was analyzed by immunohistochemistry assay ,and the relationship between the expression of livin and caspase-3 and kidney injury was analyzed .Results Compared to the control group , the rats injected endotoxin had the performance of AKI ,with obviously pathomorphological damage in the kidney tissues ,signifi-cantly increasing in the serum creatinine and urea nitrogen (P<0 .01) .The livin and caspase-3 of kidney tissues in rats with AKI caused by endotoxin were positive expression (P<0 .01) .With the lapse of time ,the trend of increasing of caspase-3 showed gradu-ally slow after the highest expression of livin .Conclusion The livin involved in the pathogenesis of endotoxin-induced AKI ,it might relieve the kidney injury and protect renal function by inhibiting casepase-3 important apoptotic effector protein .

Objective To improve early diagnosis of testicular torsion,reduce misdiagnosis and reduce the rate of orchiectomy.Methods The diagnosis and treatment of testicular torsion in 31 cases were retrospectively ana-lyzed.Results 54.8% misdiagnosis rate was in this group,all the 31 cases were diagnosed by color Doppler ultra-sound,including 19 cases of retained testicle and orchiectomy in 12 cases (38.7%).In 19 cases of this group retained testis,testicular torsion time within 5 hours was 2 cases,and postoperative had testicles survival.In the 6 cases of testicular torsion time within 5 hours to 10 hours,5 cases had the testis survival.In the 5 cases of torsion of testis time was 10 hours to 24 hours,3 caseshad the testis survival.In the 6 cases of testicular torsion time more than 24 hours,2 cases of testis survival.After postoperative following-up,19 cases of retained testis had no recurrence,all the 31 cases were found no contralateral testicular torsion and all cases sex hormone levels were in the normal levels. Conclusion Testicular torsion is easily misdiagnosed,color doppler ultrasound should be preferred.Early diagnosis and aggressive surgical exploration,unity and strictly control the orchiectomy surgery indications,means a lot to reduce the rate of orchiectomy.

Objective To investigate the impact of Ang-1 on the septic mice′pulmonary vascular endothelial barrier function and VE-cadherin and its mechanism. Methods 80 BALB/c mice were randomly divided into NS, LPS, LPS+Ang-1, LPS+Ang-1+ Ly and Ang-1 groups (n = 16). Measure VE-cadherin, Ang-2 levels in plasma and lung permeability index (LPI).Test the total VE-cadherin of lung and the phosphorylation of VE-cadherin expression. Results Plasma Ang-2 was higher compared with NS group(P<0.01) except Ang-1 group. In LPS+Ang-1 group and LPS+Ang-1+Ly group, plasma Ang-2 was lower compared with LPS group (P <0.05). In LPS+Ang-1+Ly group, plasma Ang-2 was higher compared with LPS+Ang-1 group (P<0.01). LPI, plasma VE-cadherin and lung phosphorylation of VE-cadherin were the same with the trends of the plasma Ang-2 , but the lung total VE-cadherin showed the opposite tendency. Conclusion Through the PI3K/Akt signal transduction pathway , Ang-1 may regulate septic mice′VE-cadherin , hence the pulmonary vascular endothelial barrier function improved.

Objective To investigate the serum concentration of 25-hydroxyl vitamin D in the common autoimmune diseases and whether the differences of the 25-hydroxyl vitamin D level exist in different autoimmune diseases.Methods 137 cases of autoim-mune diseases,including 71 cases of rheumatoid arthritis(RA),36 cases of systemic lupus erythematosus(SLE),16 cases of Sjogren syndrome(SS)and 14 cases of ankylosing spondylitis(AS),in our hospital from January 2012 to April 2013 were selected as the re-spondents.The serum samples were collected for detecting the 25-hydroxyl vitamin D level by the electrochemiluminescence method and comparing the differences of the 25-hydroxyl vitamin D level among different autoimmune diseases.At the same time whether the differences in the proportion of the normal level,insufficiency and lack of 25-hydroxyl vitamin D exist among different kinds of disease.Results The serum 25-hydroxyl vitamin D concentration had statistically significant difference among the patients with dif-ferent autoimmune diseases(P =0.006),which in the RA group was significantly higher than that in the other groups;the propor-tion of 25-hydroxyl vitamin D insufficiency in RA,SLE,SS and AS were 29.6%,52.8%,62.5% and 57.1% respectively,which in the SLE,SS and AS groups was significantly higher than that in the RA group(P <0.05).Conclusion The 25-hydroxyl vitamin)D insufficiency is general in common autoimmune diseases,the vitamin D supplements needs to be strengthened.

Objective To investigate the different diagnostic value of serum Chitinase 3-like 1 protein(CHI3L1)and Alpha-fe-toprotein(AFP)in diagnosing hepatocellular carcinoma (HCC).Methods One hundreds HCC patients confirmed by histopa-thology were recruited between December,2015 to April,2016 from the First Affiliated Hospital of Nanjing Medical Univer-sity.Simultaneously,100 patients with chronic hepatitis B and 59 healthy individuals,matched by sex and age with HCC pa-tients,were recruited as control groups.Serum CHI3L1and AFP were measured in different groups and the difference were analyzed by STATA 1 2.0 Statistical software.The ability of these two items in differentiating different group was analyzing by ROC curve using MedCal Ver 15.2.2 software.Results Serum CHI3L1 were significantly differences in the three groups using Kruskal-Wallis analysis (χ2=93.19,P=0.000),the differences were further compared in different two groups using Mann-Whitney analysis.The results showed that serum CHI3L1 in HCC group were significantly higher than in chronic hepatitis B and healthy control group (P=0.000,z=8.766,7.400).Serum AFP were significantly differences in the three groups using Kruskal-Wallis analysis (χ2=147.54,P=0.000),and the differences were further compared in different two groups using Mann-Whitney analysis.The results showed that serum AFP in HCC group were significantly higher than in chronic hepatitis B and healthy control group (P=0.000,z=10.938,9.033).The ROC curve analysis of serum CHI3L1 and AFP for differentiating HCC group from CHB group showed that CHI3L1 yield AUC of 0.859 (95% CI:0.803~0.904) with 85% sensitivity,79% specificity and 76.8 pg/ml cut-off value,AFP yield AUC of 0.948 (95% CI:0.904~0.974)with 85% sensitivity,98% specificity and 7.6 ng/ml cut-off value,in distinguishing HCC with CHB group,the power of AFP was superior to that of CHI3L1 (P=0.006).The ROC curve analysis of serum CHI3L1 and AFP for differentiating HCC group from healthy individuals group showed that CHI3L1 yield AUC of 0.852 (95% CI:0.787~0.903)with 85% sensi-tivity,76% specificity and 76.8 pg/ml cut-off value,AFP yield AUC of 0.929 (95% CI:0.878~0.964)with 84% sensitivi-ty,100% specificity and 7.8 ng/ml cut-off value,in distinguishing HCC with healthy individuals group,the power of AFP was also superior to that of CHI3L1 (P=0.045).Conclusion Serum CHI3L1 similar to AFP has much power ability to di-agnosis HCC,but AFP was superior to CHI3L1.

Objective To establish the reference interval for serum prostate-specific antigen (PSA) in apparent healthy men of different ages in Nanjing.Methods A total of 25 820 healthy men undergoing routine physical examinations in the First Affiliated Hospital of Nanjing Medical University from October 2013 to September 2015 were selected for the study.All of them were screened by prostate B ultrasound,excluding abnormal urinary tract diseases.The concentration of serum PSA and free prostate-specific antigen (fPSA) were measured by automatic luminescence immunoassay analyzer,and the fPSA/PSA values were calculated.The participants were divided into four groups (20～ 39,40～ 59,60～ 79 and older than 80 years old groups),then the median,5th,25th,75th and 95th percentiles of both PSA and fPSA/PSA were counted,respectively.Results The median of PSA (95th percentile ranges) of these groups by age from low to high were 0.78 (1.93),0.90 (2.93),1.34(6.60) and 2.01(11.91),respectively.The 25th to 75th percentiles were 0.55～1.11,0.61～1.36,0.77～2.51 and 0.94 ～ 4.19,respectively.The median of fPSA/PSA (95 th percentile ranges) were 0.37 (0.60),0.31 (0.56),0.28 (0.53) and 0.29(0.52),respectively.The 25th to 75th percentiles were 0.28～0.46,0.23～0.40,0.22～0.36 and 0.23～ 0.37,respectively.Among all the groups,median differences of both PSA and fPSA/PSA were statistically significant (P＜0.05),and PSA levels rise with age.PSA levels in different regions were different.Conclusion The PSA level of men under 40 years in Nanjing should be 0～2.5 ng/ml,40～60 years should be 0～4 ng/ml,while men who are above 60 years,could use 0～5 ng/ml as reference interval.

Objective:To assess the prognostic value of methylation of interferon regulatory factor 6 ( IRF6) gene promoter in patients diagnosed with Kidney renal clear cell carcinoma (KIRC). Methods:The primary lesions of fifty KIRC patients who were diagnosed at the First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. The expression of IRF6 protein was determined with an immunohistochemical method. The correlation between the level of IRF6 expression and survival and/or metastasis status was analyzed. The mRNA and protein levels of the IRF6 in KIRC and normal renal tissues were compared by using bioinformatic tools. The difference in the methylation rate of the IRF6 gene promoter between tumor and adjacent tissues was analyzed by searching the online databases. Statistical analysis was carried out for the methylation status of the IRF6 gene promoter region to select those negatively correlated with the overall survival (OS) among the patients. In vitro experiments were conducted with cell lines to verify the correlation between the status of promoter methylation and transcription level of the IRF6 gene. Results:The mRNA and protein levels of the IRF6 gene in KIRC tissues were significantly lower than those of the normal controls, and this was more prominent in patients who had died or developed metastasis. The extent of IRF6 gene promoter methylation in the KIRC tissues was much higher compared with that of the adjacent normal renal tissues. There was a significant negative correlation between the methylation of the IRF6 gene promoter and mRNA level of the IRF6 ( R= -0.52). The higher methylation degree in the IRF6 gene promoter regions cg12034118 and cg16030177, the shorter the OS and worse prognosis in the patients. Only twenty CpG sites in cg12034118 were confirmed to be highly methylated in KIRC cell lines. The transcription level of the IRF6 gene was upregulated in a time- and dose-dependent manner after the treatment with demethylation reagent 5-azadeoxycytidine. Conclusion:The methylation of IRF6 gene promoter in the renal tissues of KIRC patients is closely correlated with the OS. Cg12034118 may provide a promising biomarker for laboratory detection, and its high methylation rate has certain reference value for the prognosis.