Abstract:Objective:To explore whether Cdk4 and CyclinD3 are involved in the inhibition of β-cell proliferation caused by miRNA-24. Methods:miRNA-24(miR-24)was highly expressed in MIN6 cells. Cell viability was determined using WST-1 assays. Cells were collected for flow cytometry analysis to test cell cycle distribution. Protein levels of Cdk4 and CyclinD3 were analyzed by Western blot. CyclinD3-pCMV5-myc and cdk4-pCMV5-myc recombinant plasmid were constructed and transfected into MIN6 cells together with pre-miR-24 or pre-Neg miRNA precursors. Cell proliferation was measured by the BrdU labeling. Results:MIN6 cells proliferation was reduced because of transfected miR-24,mainly due to G2 phase arrest. No changes of the level of apoptotic cells were detected. Cdk4 and CyclinD3 were downregulated by overexpressed miR-24. Overexpressed cdk4 and cyclinD3 reversed the β-cell proliferation inhibition. Conclusion:miR-24 inhibits β-cell proliferation by decreasing Cdk4 and CyclinD3 protein levels. The inhibition may be reversed by overexpressed cdk4 and cyclinD3.

Abstract:Objective:To investigate the expressions and significances of mTORC1-HIF1α pathway genes in human CD8+ regulatory T cell. Methods:Coculture systerm of CD8+ T cells and SKOV3 were conducted in vitro,CD8+ T cells cultured alone group acted as control group. At day 5,CD8+ T cells were collected,and used to examine the expression of eight glycolysis genes by quantitative real-time reverse transcriptase(qRT)-PCR(mTORC1,HIF1α,Glut1,PKM2,GPI,TPI,Eno1,and LDHα). Western blot was used to detect the expression of mTORC1,HIF1α and PKM2. We collected peripheral blood from 14 ovarian cancer(OC) patients,12 benign diseases,and 12 healthy females. CD8+ T cells were then separated using a CD8-positive isolation kit. The expressions of mTORC1,HIF1α,Glut1,PKM2,GPI,TPI,Eno1,and LDHα in CD8+ T cells from OC patients was detected by qRT-PCR and compared with that in patients with ovary benign tumor(BOT)and healthy volunteers. Results:Compared with CD8+ T cells cultured without SKOV3 cells,glycolysis gene expression showed varying degrees of decline in CD8+ T cells cultured with SKOV3 cells. The expression of mTORC1,HIF1α,PKM2,GPI,and TPI was significantly decreased in co-cultured cells compared with the control group. Results showed that the expression of mTORC1,HIF1α,and PKM2 decreased significantly(P < 0.05)in CD8+ T cells cultured with SKOV3 cells compared with control group. HIF1α and Glut1 mRNA had lower expression levels in CD8+ T cells from OC patients than those from healthy controls(both P < 0.05). mTORC1,PKM2,GPI,and TPI were also expressed at lower levels in OC patients than those in either BOT patients or healthy controls(both P < 0.05). The expression of mTORC1-HIF1α pathway genes has no difference between benign diseases group and the healthy controls. Conclusion:mTORC1-HIF1α signalling integrates the control of CD8+ T cells metabolism and differentiation in ovarian cancer microenvironment,which play a significant role in the immunotherapy of OC

Abstract:Objective:To investigate the migration and invasion of lung neoplasm potentiated by MKL1 and the change of CCNB2 activity involved in signal pathway. Methods:Lung cancer cells were infected with lentivirus carrying indicated shRNAs and effect of MKL1 inhibition on migration and invasion of lung cancer cells was measured. Small interfering RNA was transfected to suppress the expression of MKL1,RT-PCR was performed to examine CCNB2 mRNA level,and luciferase reporter assays was to assess the effect of MKL1 deletion on the promoter activities of CCNB2. Results:Cell colony formation was diminished by soft agar clonogenic assay and cell proliferation was reduced when MKL1 was silenced by siRNA. MKL1 depletion directly repressed CCNB2 transcription,and the inhibition was up to 70%. RT-PCR showed great depression of CCNB2 mRNA by the interference of MKL1. Difference was of significance(P < 0.05). Conclusion:Depletion of MKL1 by shRNA significantly hampered the migration and invasion of lung cancer cell. MKL1 depletion inhibited CCNB2 transcriptional activity and repressed mRNA expression. The study provided early-stage markers for lung cancer diagnosis and treatment.

Abstract:Objective:To investigate the effect of titanium modified by anodic oxidized TiO2 nanotubes on the osteogenic differentiation of human periodontal ligament stem cells(hPDLSCs). Methods:TiO2 nanotubes group and control group(smooth titanium)were involved in this study. The cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay. The osteogenic differentiation was evaluated using alkaline phosphatase(ALP)activity assay. In addition,osteogenic marker genes expression including Col-I,OCN and Runx-2 was evaluated. Results:We prepared nominally TiO2 nanotubes with 100 nm diameter on Ti substrates by anodization. No significant difference was detected on the proliferative activity at day 1 and the ALP activity at day 4 between the TiO2 nanotubes group and control group(P > 0.05). However,cell proliferative activity of TiO2 nanotubes group was lower than that of the control group at day 4 and 7. ALP activity assay demonstrated that the TiO2 nanotubes group at day 7,14 and 21 had a high ALP activity. The gene expression of Col-1,OCN,and Runx-2 of TiO2 nanotubes group was up-regulated at 21-days. Conclusion:The proliferation activity of hPDLSCs on the TiO2 nanotubes was less than that of control group. ALP activity,gene expression TiO2 nanotubes group were higher than those of on control group,and it suggested that TiO2 nanotubes could promote osteogenic differentiation of hPDLSCs.

Abstract:Objective:To investigate the emulsified isoflurane post-conditioning on isolated lung ischemia-reperfusion(I/R)injury in rats. Methods:Sprague-Dawley rats were randomly divided into four groups(n=6 each):control group(group S),ischemia-reperfusion group(group IR),emulsified isoflurane group(group EI)and intralipid group(group IL). Lung ischemia-reperfusion injury model was established. Recorded tidal volume(Tidal volume,VT),pulmonary venous partial pressure of oxygen(Oxygen partial,PaO2),lung compliance(Compliance,CL)and airway resistance(Airway resistance,R)values at the end of the balance(T0),immediately prior to ischemia and hypoxia(T1),reperfusion 30 min(T2)and reperfusion 60 min (T3);measured lung wet / dry weight ratio (W/D);observed lung tissue HE staining and detected the expression levels of SOD and MDA. Results:Compared with T0 moment,VT,CL and PaO2 decreased but R increased at T3 moment in the four groups(P < 0.05 or 0.01). Compared with S group,VT,CL and PaO2 significantly decreased but R obviously increased at T2-3 moment in IR group and IL group(P < 0.05 or 0.01). Compared with IR group,VT,CL and PaO22 increased but R decreased at T2-3 moment in EI group and IL group(P < 0.05 or 0.01). Compared with EI group,PaO2 decreased at T2-3 moment,VT and CL decreased but R increased at T3 moment in IL group(P < 0.05). Compared with S group,W/D,the expression level of MDA increased and the expression level of SOD decreased in IR group and IL group(P < 0.05 or 0.01),the expression level of SOD decreased in EI group(P < 0.01). Compared with IR group,W/D,the expression level of MDA decreased and the expression level of SOD increased in EI group(P < 0.05 or 0.01),W/D decreased and the expression level of SOD increased in IL group(P < 0.05). Compared with EI group,the expression level of SOD decreased and the expression level of MDA increased in IL group(P < 0.05). Pathology showed that the damage of lung tissue of group IL or group EI was lighter than that in group IR,the damage of lung tissue of group EI was lighter than that in group IL. Conclusion:8% emulsified isoflurane post-conditioning can prevent isolated lung ischemia-reperfusion injury in rats,and the mechanism may be related to alleviating lung oxidative stress caused by ischemia-reperfusion injury.

Abstract:Objective:To investigaee the expressions of VEGF and EGFR and their associations with the clinical biological behavior in primary liver cancer. Methods:The protein expressions of VEGF and EGFR were detected in 63 primary liver cancer by immunohistochemistry streptavidin-perosidase(SP)methods. Results:①The positive expression rate of VEGF and EGFR was 63.5% and 50.8%,respectively.②The expression of VEGF was significantly correlated with Edmondson pathologic grade,portal vein tumor thrombus,tumor capsule and clinical stage(P < 0.05),while not with sex,age,AFP level,HBsAg status,liver cirrhosis,histological type and tumor size(P > 0.05).③EGFR expression was related to histological type,Edmondson pathologic grade,tumor mass and tumor capsule(P < 0.05). No correlation was found between EGFR expression and sex,age,AFP level,HBsAg status,liver cirrhosis,portal vein tumor thrombus,clinical stage(P > 0.05). Conclusion:VEGF and EGFR play an important role in the growth and metastasis of primary liver cancer.

Abstract:Objective:The second generation intact parathyroid hormone (iPTH) assay is the most widely used method for the measurement of PTH which detect both full-length(1~84)PTH and(7~84)PTH. Whereas,the third generation whole PTH(wPTH)assay has been demonstrated to be specific for the measurement of(1~84)PTH with high attention from the clinic. This research was aimed to explore the features of intraoperative levels of plasma iPTH,(1~84)PTH,(7~84)PTH in chronic kidney disease complicated with secondary hyperparathyroidism(SHPT) patients with parathyroidectomy(PTX). Methods:Intraoperative levels of plasma iPTH,(1~84)PTH,(7~84)PTH in 58 severe SHPT patients who received total PTX+autotransplantation were recorded. Plasma iPTH and(1~84)PTH levels were measured by the second and third generation assays,respectively. Circulating(7~84)PTH levels were calculated by subtracting the(1~4)PTH values from the iPTH values. Results:Plasma iPTH,(1~84)PTH,(7~84)PTH levels were closely associated with each other(P < 0.001) at baseline,10 and 20 minutes after removing the last parathyroid gland. After PTX,plasma levels of iPTH,(1~84)PTH,(7~84)PTH had dropped to(19.3 ± 9.5)%,(12.7 ± 8.1)%,(26.7 ± 12.1)%,respectively at 10 minutes,(14.1 ± 5.9)%,(7.9 ± 4.4)%,(20.9 ± 8.5)% at 20 minutes respeatively. Conclusion:The third generation PTH assays were closely correlated with the second generation assays. PTX can significantly abate the levels of plasma different PTH fragments in severe SHPT patients,and intraoperative concentration of plasma(1~84)PTH was decreased more rapidly than iPTH and(7~84)PTH.

Abstract:Objective:To investigate the clinical utility of carbon nanoparticles suspension as lymph node tracer in the cervical lymphadenectomy and protection function of parathyroid in patients with papillary thyroid cancer. Methods:One hundred twenty consecutive patients were divided into carbon nanoparticles suspension group(n=60) and control group(n=60) for prospective studies. All patients underwent total thyroidectomy with unilateral cervical lymphadenectomy. The operating time,operative blood loss,postoperative drainage volume,the numbers of parathyroid autotransplantation,the total number of cleaning lymph nodes(cN0 and cN1) and the diameter < 5 mm lymph node,and metastasis lymph nodes,the parathyroid hormone(PTH),the serum calcium level,and the cases of hypocalcemia after surgery were compared between the two groups. Results:The operating time was longer than that of the control group(P=0.002),operative blood loss and postoperative drainage volume had no statistically significant difference between the two groups(P > 0.05). Carbon nanoparticles suspension group cleaned out 519 pieces of lymph nodes(average 10.6),was compared with the control 293 pieces of lymph nodes(average 6.9)(P=0.000 1). In cN0 cases,the total numbers of lymph node cleaning and diameter < 5 mm lymph node with carbon nanoparticles suspension were obviously more than the control group(P=0.000 1),but in the cN1 cases,there was no difference on the total number of lymph nodes(P=0.086) between the two groups,yet it was helpful for the diameter < 5 mm lymph node inspection(P=0.000 1). Postoperative hypocalcemia of the controls was more than the carbon nanoparticles suspension group(P=0.001). Conclusion:Carbon nanoparticles suspension might help to detect metastatic lymph nodes with thyroid cancer cN0,especially diameter < 5 mm lymph node inspection,and protect the function of the parathyroid gland.

Abstract:Objective:To study the change characteristics of muscle mass,fat mass,moisture,basal metabolic rate,and bone mineral density of patients with colorectal cancer. Methods: 198 patients with colorectal cancer and 129 healthy persons were carried on the physical anthropology analysis using by body composition analyzer or bone mineral density instrument. Results: Weight,abdomen circumference,body mass index(BMI),systemic fat mass,trunk fat mass,and intracellular fluid,and bone density of patients with colorectal cancer were less than those in the control group (P < 0.05),while the extracellular fluid and basal metabolic rate were higher than those in the control group (P < 0.05). There were group differerces among the fat mass,intracellular fluid and extracellular fluid of patients with differrent pathological stage (P < 0.05). Conclusion: The changes of body composition in patients with colorectal cancerr were obvious,accompanying the decrease of fat,intracellular fluid and bone mineral density,while the increase of extracellular fluid and basal metabolic rate.

Abstract:Objective:To explore the risk factors of bloodstream infection in patients with hydrocarbon resistant Klebsiella pneumoniae(CRKP)and the risk factors of death in patients with Klebsiella pneumoniae bloodstream infection. Methods:Clinical data from 91 patients with Klebsiella pneumonia bloodstream infection were analyzed in the Second Affiliated Hospital of Nanjing Medical University from December 2009 to January 2014. The risk factors of CRKP were determined by the chi square test,and the risk factors for the death of Klebsiella pneumoniae bloodstream infection were determined by Logistic regression. Results:Im CRKP group,proportion of bloodstream infectied patients complicated by pneumonia(P=0.016),invasive mechanical ventilation(P=0.048)and ICU(P=0.017) was significantly higher than that im the SKP group and ESBLKP group. Univariate logistic regression analysis showed that WIC score≥3,malignant tumor,renal function imsufficiency,hypoproteinemia,deep vein set tube,with invasive mechanical ventilation,ICU admission,Pitt bacteremia score≥6 were the risk factors of death in patients with Klebsiella pneumoniae bloodstream infection. Multivariate logistic regression analysis showed that Pitt bacteremia score≥6 points(OR=509.891,P < 0.001)was an independent risk factor of patients died of Klebsiella pneumoniae bloodstream infection. Conclusion:With pneumonia,invasive mechanical ventilation and ICU stay were risk factors for CRKP-producing bloodstream infection;Pitt bacteremia score≥6 was the independent risk factor of patients died of Klebsiella pneumoniae bloodstream infection.

Abstract:Objective:To investigate the value of lung ultrasonic score(LUS)for quantitative ultrasound diagnosis in neonatal in the neonates with acute respiratory distress syndrome(ARDS). Methods:Retrospectively,these participants with the suspected ARDS were considered candidates for the prospective trial between January 2012 and January 2016 in the First Affiliated Hospital of Nanjing Medical University neonatal intensive care unit,and 90 cases were included according to the diagnostic criteria of ARDS in 2012 Berlin. Lung ultrasound examination and lung functions were monitored for evaluation of the neonates with ARDS. Results:The LUS of term ARDS group and premature ARDS group was(15.6~3.9)and(20.5~4.2),respectively,with statistically significant difference (P < 0.05). Conclusion:Bedside lung ultrasound has a better accuracy in diagnosis of neonatal ARDS,which deserves to be popularized in the clinical practice.

Abstract:目的:探讨血尿酸(uric acid,UA)水平-血尿酸肌酐比(uric acid/creatinine ratios,UA/Cr)与帕金森病(Parkinson’s disease,PD)运动分型及病情的相关性-方法:将2014—2015年本院住院的PD患者77例为实验组,另77例性别-年龄匹配的健康体检者为对照组,比较两组UA及UA/Cr水平的差异-将实验组依照统一帕金森病评定量表(UPDRS)分为两组,其中以震颤为主型(tremor dominant,TD)26例,姿势异常步态障碍为主型(postural instability gait difficulty,PIGD)51例,比较两组UA及UA/Cr水平的差异-结果:实验组UA及UA/Cr显著低于对照组(P < 0.000 1),PIGD组UA及UA/Cr显著低于TD组-结论:UA及UA/Cr是PD的危险因素,同时也可作为其区分运动分型的生物学指标-

Abstract:Objective:To investigate and improve the management of type 1 diabetes mellitus in Jiangsu Province. Methods:A questionnaire designed for this survey was used to learn about the diagnosis,treatment,glycemic control,education and follow-up of type 1 diabetes mellitus among endocrinologists from 35 tertiary hospitals in Jiangsu Province. Results:Altogether 417 valid questionnaires were collected. Among them,71.9% of endocrinologists preferred multiple daily injections basal-bolus insulin regimens from diagnosis,and 75.1% of endocrinologists had prescribed oral medicines in combination with insulin. There was no significant difference among different professional title groups(P > 0.05). Self-monitoring of blood glucose(SMBG)and hemoglobin A1C measurement were recommended by 91.4% and 85.6% of endocrinologists respectively. Hypoglycemic events and ketosis occurred in nearly 80% patients. The self-management skills had become the main factor influencing endocrinologists’ decisions.Conclusion:The mastering of basic perception of type 1 diabetes mellitus knowledge is satisfactory among the endocrinologists,while overall level of diagnosis and treatment remains to be improved.

Abstract:Objective:The mortality and PYLL of malignancies in Jiangsu province in 2013 were studied to provide basic data for cancer prevention and control. Methods:Death case information related to malignancies and demographic information was collected through Provincial Death Cause Surveillance System and police department,disease burden of malignancies and its population distributive characteristics were analyzed by calculating mortality,standardized mortality,PYLL and SPYLL,etc. Results:Totally 135 236 cases were died of malignant tumor,which accounted for 29.38%. Arnong the sabjects,86 736 cases were male and 48 500 cases were female. The crude and standardized mortality were 196.80/100 000 and 111.59/100 000,respectinely the male to female ratio of crude mortality was 1.77,while the urban to rural ratio was 0.95,and the top five were lung cancer,stomach cancer,esophageal cancer,liver cancer,and colorectal cancer,which accounted for 75.54% of all. PYLL and SPYLL was 788 367 and 582 620.68 people*years,respectively,the male to female ratio of PYLL was 1.78,while the urban to rural ratio was 0.93. Liver cancer,lung cancer,stomach cancer,esophagus cancer and leukemia were the top five,however,breast cancer had replaced the fourth place of esophagus cancer in the female. Conclusion:The leading malignancies related to death and early death in the whole population were lung cancer,gastric cancer,esophageal cancer,liver cancer,colorectal cancer and leukemia,as well as breast cancer in the female. Malignant tumor mortality and PYLLR of the male was much higher than those of the female,and those of the rural residents was slightly higher than those the urban of. Preventive interventions should focus on key population and malignancy with higher mortality burden.

Abstract:Objective:To estimate the risk of deaths attributable to carbapenem-resistant enterobacteriaceae by performing a systematic review and meta-analysis. Methods:The Embase and PubMed databases were searched from January 1,2001 to December 31,2015 to identify eligible cohort studies. The initial literatures and references listed in the literature were manually searched. Controlled studies were analyzed using stata14.0 software. The risk ratio(RR)was pooled and the publication bias was evaluated using funnel plots. Results:Sixteen studies with a total of 2 916 patients were included for systematic review(the number of patients with carbapenem-resistant infections were 797 and those with carbapenem-susceptible infections were 2 119). According to the type of design,studies were divided into cohort studies and case control studies. According to the infection of specimen type,studies were divided into bloodstream infection type subgroup and comprehensive infection type subgroup. The risk ratio of the mortality in cohort studies was 2.44(95%CI:1.96~3.03,I2=16.1%,Pheterogeneity=0.312);the risk ratio of bloodstream infection was 2.07(95%CI:1.59~2.69,I2=0.0%,Pheterogeneity=0.511);the risk ratio of comprehensive infection type subgroups was 3.35(95%CI:2.25~4.99,I2=0.0%,Pheterogeneity=0.868). The rodds ratio of the mortality in case control studies was 2.30(95%CI:1.49~3.55,I2=53.5%,Pheterogeneity=0.018). The rodds ratio of bloodstream infection was 2.85(95%CI:1.66~4.90,I2=60.8%,Pheterogeneity=0.018). The rodds ratio of Comprehensive infection type subgroups was 1.46(95%CI:0.73~2.90,I2=25.2%,Pheterogeneity=0.261). Conclusion:Pooled outcomes showed that the number of deaths was significantly higher in patients with carbapenem-resistant infections and that the number of deaths attributable to carbapenem resistance is considerable,carbapenem resistant enterobacteriaceae in the bloodstream infections is closely related to death of patients.