Abstract:Objective：To explore the regulatory role of KLF4，as a transcription factor，in the production of pro-inflammation cytokine IL-23 gene in rat glomerular mesangial cells（GMC）induced by sublytic C5b-9. Methods：The plasmids of KLF4 short hairpin RNA（shKLF4）and KLF4 overexpression（pIRES2-KLF4）were first generated and transfected rat GMC respectively，then the cells were treated with or without sublytic C5b-9. The roles of silencing KLF4 gene in IL-23 synthesis induced by sublytic C5b-9 or overexpressing KLF4 gene in IL-23 production were detected by qPCR，Western blot and ELISA. Moreover，the plasmid of IL-23 promoter（full-length，FL）was also constructed. After co-transfection，the activity of IL-23 promoter with or without sublytic C5b-9 after KLF4 gene knockdown or overexpression was measured by luciferase reporter assay. Results：①Sublytic C5b-9 could obviously elevate IL-23 production in rat GMC，and after KLF4 gene knochdown with shKLF4，the increase of IL-23 expression upon sublytic C5b-9 was remarkably diminished，but KLF4 overexpression in the GMC could markedly elevate IL-23 synthesis. ②The IL-23 promoter activity incubated with sublytic C5b-9 was significantly up-regulated. Meanwhile，after KLF4 gene knockdown，the IL-23 promoter activity in the GMC stimulated by sublytic C5b-9 was obviously declined，but KLF4 overexpression greatly increased IL-23 promoter activivy. Conclusion：Sublytic C5b-9 could induce IL-23 production of GMCs through the increase of KLF4 expression.

Abstract:Objective：To explore the effects of nuclear localization sequence（NLS） and carboxyl terminus of parathyroid hormone related peptide（PTHrP） on the oligodendrocytes development and myelination in mice. Methods：PTHrP knock in（PTHrP KI）mice and their wild type（WT） littermates were accepted 5-bromo-2-deoxyuridine（BrdU） intraperitoneal injection at postnatal day 6（P6），and were sacrificed at the next day. The myelination in the central nervous system（CNS） was observed by myelin basic protein（MBP）immunohistochemical staining and electron microscope. The proliferation of oligodendrocyte progenitor cells in the hippocampus was detected by BrdU labeling and immunofluorescence staining. O4 positive oligodendrocyte progenitor cells（OPCs）from the cerebral cortex of P5 PTHrP KI mice and their WT littermates were selected and cultured in vitro. Then，the Ki67 positive cells were observed by immunofluorescence staining，the ratio of apoptotic cells was measured by flow cytometer analysis，and the proliferation and apoptosis related proteins were detected by Western blot. The differentiation ability of OPCs was inspected by 2′，3′-cyclic-nucleotide 3′-phosphodiesterase（CNPase）immunofluorescence staining after cultured in differentiation medium for 7 days. Results：Comparison to their WT littermates，PTHrP KI mice showed decreased expression of MBP positive nerve fibers and the thinner myelin sheathes of axons in the brain，decreased BrdU and oligodendrocyte transcription factor 2（Oligo2）double positive cells in hippocampus area. The in vitro cultured OPCs from PTHrP KI mice showed the decreased ratio of Ki67 positive cells，the decreased expression level of proliferation related proteins as B cell-specific MLV integration site-1（Bmi-1）and proliferating cell nuclear antigen（PCNA），the increased ratio of AV+/PI- apoptotic cells，the increased expression level of apoptotic proteins as caspase-3 and p16，and the decreased intensity of CNPase immunofluorescence after 7 days’ differentiation culture. Conclusion：Defect of NLS and C-Terminus of PTHrP leads to the decreased proliferation，increased apoptosis，and delayed differentiation of OPCs，and results in the axonal myelination disability in CNS.

Abstract:Objective：To screen B cell and T cell antigen epitopes of Acinetobacter baumannii protein TonB. Methods：B cell epitopes and T cell epitopes were predicted by bioinformatics methods and synthesized. Recombinant pET28a-tonB was cloned and TonB was purified to immunize mice. BALB/c mice were immunized by subcutaneous for 3 times. Five weeks later，serum was collected from each of immunized mice and then negative control were mice used to perform indirect ELISA with synthesized peptides to identify B cell epitopes. Cells of the spleen were separated from every mouse and cultured with each of synthesized peptides respectively，and then cell supernate was collected to detect IFN-γ secretion by double sandwich ELISA for T cell epitopes selection. Results：Construction of 4 candidate B cell and T cell epitopes was performed，including PB1，PB2，PB3，PB4，PT1，PT2，PT3，and PT4，respectively. Indirect ELISA showed that peptide PB3，PB4 reacted with immunized mice serum and the A450 nm value was significant higher than that of the negative mice（P＜0.05）. Double sandwich ELISA showed that IFN-γ concentration that stimulated by peptide PT2，PT3，and PT4 was significant higher than its negative control group（P＜0.05）. Conclusion：Two B cell epitopes PB3 and PB4，3 T cell epitopes PT2，PT3，PT4 of TonB were successfully constructed and screened.

Abstract:Objective：To obtain a set of aptamers of liver cancer serum markers by SELEX screening，and to provide new molecular biological detection method for the early diagnosis of liver cancer. Methods：Serum samples from 50 patients with primary hepatocellular carcinoma（HCC） and 50 normal serum samples of which physical examination showed no abnormalities were collected and mixed in equal proportions to prepare mixed sera of HCC and normal subjects. The serum was used as the target molecule for screening. Magnetic beads-normal human serum complex was combined with ssDNA library. The supernatant was then combined with magnetic beads-liver cancer serum to elute and isolate the specific ssDNA binding to serum of liver cancer. Nine rounds of ssDNA were screened by the method of the streptavidin biotin method，and transformed into PMD18-T vector. The single clones were selected and sequenced by Shanghai Biosystems. Affinity determination of serum tumor marker aptamers in patients was performed by flow cytometry. Results：After 9 rounds of screening，200 nucleic acid sequences were successfully isolated，of which 10 sequences were different. Specificity test showed that the binding dissociation constants of serum tumor marker aptamers and HCC serum were all in nanomolar level. Among them，Seq-1，Seq-16，Seq-17，Seq-56，Seq-72 aptamers could be bound to the serum of liver cancer with highly specificity，and could not bound to the normal serum. The positive rate of serum tumor marker aptamers was detected in 200 cases of liver cancer serum and 200 cases of normal human serum，positive detection rate was more than 91%. Conclusion：The use of a random single-stranded oligonucleotide library to successfully obtain an aptamer that specifically binds to liver cancer patient serum，and the obtained aptamer has the ability to antagonize liver cancer serum，which may provide a new convenient method for the early diagnosis of liver cancer.

Abstract:Objective：To investigate effects of 17β-estradiol（E2）on mechanical stretch-induced integrin β1/FAK/p38 MAPK signal pathway of cardiomyocyte hypertrophy. Methods：Cardiomyocyte from neonatal rats were cultured in vitro and cardiomyocyte hypertrophy was induced by mechanical stretch. The association of integrin β1 and FAK was evaluated by immunoprecipitation，and the phosphorylation of FAK and p38MAPK were determined by Western blot. Results：Mechanical stretch for 24 h significantly increased the association of integrin β1 and FAK，and increased the phosphorylation of FAK and p38MAPK. Pretreatment with 100 nmol/L E2 for 30 min significantly attenuated the increases in the association of integrin β1 and FAK which were attenuated by estrogen receptor antagonist ICI182780. Furthermore，E2 also decreased the levels of the phosphorylation of FAK and p38MAPK induced by mechanical stretch，which were also attenuated by ICI182780. Conclusion：The combination of E2 and estrogen receptors can inhibit the activation of integrin β1/FAK/p38MAPK signaling pathway induced by mechanical stretch to play a cardiovascular protective effect.

Abstract:Objective：To detect the expression and function of Kv1.3 channel in peripheral blood T lymphocytes of asthma patients，to study the effect of Kv1.3 channel blockers on the proliferation of T lymphocytes and the production of Th2 cytokines in peripheral blood of asthmatic patients，and to explore the possibility of Kv1.3 channel blockers in treating chronic airway inflammation in patients with asthma. Methods：T lymphocytes were isolated from peripheral blood of asthma patients and healthy controls. Then，we detected the mRNA levels of Kv1.3 in T cells. We detected the effect of Kv1.3 channel inhibitors ShK on T cell proliferation and the production of Th2 cytokines. Results：The Kv1.3 level of peripheral blood T lymphocytes in patients with asthma was significantly higher than that of healthy controls. ShK significantly inhibited the proliferation and Th2 cytokines production ability of T lymphocytes. Conclusion：Kv1.3 may play important roles in inflammation in asthma. Selective blocking of Kv1.3 channels may be one of the future treatments of asthma.

Abstract:Objective：To investigate the effects of vitamin E on airway remodeling in asthma rats. Methods：Twenty-four clean grade SD male rats were randomly divided into the control group，the asthma group and the vitamin E group，with 8 in each. The asthma group and the vitamin E group were sensitized and challenged with ovalbumin to establish asthmatic models. The control group was treated with the same amount of saline. From the 15th day，the vitamin E group was fed with vitamin E，100 mg/（kg·d）. The other two groups were fed with equal amount of saline. On day 69，all rats were sacrificed in 24 hours after challenged，then the serum and lung tissue specimen were preserved. The pathological changes of lung tissues were detected by HE staining. The thickness of airway wall and smooth muscle was detected by computer image analysis system. The expression of hypoxia inducible factor-1α（HIF-1α），matrix metalloproteinase-9（MMP-9）and tissue inhibitor of metalloproteinase-1（TIMP-1）in the serum was detected by ELISA. MMP-9 and TIMP-1 in lung tissues were observed by immunohistochemistry. Results：Compared with the control group，the thickness of airway wall and smooth muscle in the asthma group significantly increased，while the vitamin E group somehow improved in the above changes when compared with the asthma group. Compared with the control group，the expression of HIF-1α，MMP-9，TIMP-1 in serum and MMP-9，TIMP-1 in lung tissues in the asthma group significantly increased. while HIF-1α，MMP-9，TIMP-1 in serum and MMP-9，TIMP-1 in lung tissues in the vitamin E group obviously decreased when compared with the asthma group（P＜0.05）. Conclusion：Vitamin E may improve airway remodeling in asthma by inhibiting HIF-1α，MMP-9 and TIMP-1.

Abstract:Objective：To analyze pathogenic characteristics of diarrheogenic Escherichia coli（DEC）in the patients with food borne diseases in Jiangsu Province；to understand drug resistance status and provide effective basis for better prevention，control and clinical treatment in the future. Methods：The fecal specimens of diarrhea patients in 48 food borne disease sentinel hospitals in Jiangsu Province were collected in 2017，5 kinds of DEC were detected by traditional separation culture method combined with fluorescence quantitative PCR. The infection characteristics were analyzed by epidemiological description. At the same time，266 DEC strains were subjected to drug resistance test against 14 antibiotics of 8 categories by micro broth dilution method. Results：Among 9 884 samples of fecal or anal swabs，a total of 289 DEC were detected（accounted for 2.92%），the highest detection rate in 5 types was EAEC（accounted for 1.53%），followed by ETEC（accounted for 0.81%）and EPEC（accounted for 0.54%），2 samples were double bacteria combined infection. The positive DEC detection rate was relatively higher in summer，the highest rate was observed in August. The proportions of male and female infection（2.78% and 3.08%，respectively）were approximately equal and no statistical difference was found between different genders（χ2=0.801，P＞0.05）. For age distribution，the highest rate was detected among the 1 year old group（accounted for 4.55%），followed by 1-5-year-old group（accounted for 4.06%），the detection rate of different age groups was statistically significant（χ2=45.780，P < 0.01）. The DEC demonstrated high drug resistance rate（accounted for 91.35%），among which ampicillin had the highest drug resistance rate（64.3%），followed by nalidixic acid（60.9%）and tetracycline（51.9%）；multi-drug resistance rate was 60.9%，1 strains was resistant to all 14 kinds of antibiotics. Conclusion：The infection rate of EAEC in DEC is the highest among food borne DEC in Jiangsu province. Infants younger than 5 years old require emphasized focus. The multi-drug resistance of DEC is relatively serious，which needs to more attention.

Abstract:Objective：To investigate the related factors of anemia in patients with diabetic nephropathy（DN）diagnosed by renal biopsy. Methods：In this retrospective study，178 patients with DN diagnosed by renal biopsy were included. The levels of hemoglobin（Hb）<130 g/L（in males） and Hb<120 g/L（in females） were identified as anemia；The levels of Hb≥130 g/L（in males）and Hb≥120 g/L（in females）were identified as non-anemia. The relationship between anemia and clinical manifestations，laboratory findings，pathological classification of DN，renal tubular injury scores was analyzed. Logistic regression analysis was used to evaluate related factors in DN patients of different genders. Results：In 178 cases with DN，duration of diabetes（DD），estimation of glomerular filtration rate（eGFR），urinary albumin/creatinine（UACR），glycosylated hemoglobin（HbA1c），albumin（Alb），triglyceride（TG），low density lipoprotein cholesterol（LDL-C），serum iron（SI），transferrin（TF），high sensitive C reactive protein（hs-CRP）were significantly different between the anemia and the non-anemia group. There were significant differences in DD，eGFR，UACR，HbA1c，Alb，TG，SI，TF，hs-CRP between the two groups of male patients；in female cases，DD，Alb，HbA1c，UACR，hs-CRP were significantly different between the two groups. Logistic regression analysis showed that eGFR，Alb and DD，hs-CRP were related to anemia in male patients；Alb and hs-CRP were correlated with anemia in female patients. The tubulointerstitial damage score in the anemia group were significantly higher than that in the non-anemia group，and significant difference in Hb level between pathological stage Ⅱa and other stages was also detected. Conclusion：Related factors of anemia in DN patients include duration of diabetes，renal function deterioration，iron metabolism，inflammation and nutritional deficiency. Tubulointerstitial damage might be an important mechanism of renal function deterioration associated anemia.

Abstract:Objective：To investigate the clinical value and significance of high risk human papilloma virus（HPV）E6/E7 mRNA test in patients with HPV infection. Methods：Between January 2017 and June 2017，in Center of Diagnosis and Treatment for Cervical Disease，200 patients with HPV infection in cervical screening received high-risk HPV E6/E7 mRNA test and pathological biopsy under colposcopy. We analyzed the E6/E7 mRNA detection rate of positive cases，and the E6/E7mRNA expression in different pathological diagnosis. Results：The E6/E7 mRNA detection rate was 60% in the 200 patients. The highest E6/E7 mRNA positive expression in the high-grade squamous intraepithelial lesion group（HSIL）had significant differences with the low-grade squamous intraepithelial lesion group（LSIL）and the nomal group. The mRNA sensitivity was 97.9% and negative predictive value was 98.8%（P < 0.05）. The sensitivity，specificity，positive predictive value and negative predictive value were lower in the LSIL group. Conclusion：We could shunt the patients with HPV infection，avoid excessive treatment and reduce the patient’s psychological burden byHR-HPV E6/E7 mRNA test. High-risk HPV E6/E7 mRNA test has high clinical value in diagnosis of HSIL during cervical screening.

Abstract:Objective：To investigate the serum level of stanniocalcin-1（STC1） in patients with stable chronic obstructive pulmonary diseases（COPD） and to explore the relationship between STC1 and severity of the disease. Method：A total of 55 patients with definite stable COPD and 24 healthy controls were enrolled in this study. Serum STC1 was measured by enzyme-linked immunosorbent assay（ELISA），and the correlation between STC1 and COPD was analyzed. Results：①Serum STC1 levels were significantly lower in patients with COPD than healthy controls（P < 0.05）. ②The receiver-operating characteristic（ROC） curve of serum SCT1 was used for threshold identification（AUC=0.659，P=0.025），the optimal cut-off point of serum STC1 was 1 350 pg/mL（sensitivity 81.82%，specificity 50.00%，respectively）. ③Poor association of serum STC1 levels with stable COPD was found，including the GOLD classification，COPD assessment test（CAT） scores and lung functions. Conclusion：Serum STC1 levels were significantly decreased in COPD patients while it is poorly correlated with disease severity. STC1 might have clinical application in stable COPD.

Abstract:Objective：To determine the pathological significance of serum miR-21 expression level in premature respiratory distress syndrome（RDS）infants. Methods：Randomized blood samples were collected from RDS and non-RDS infants in the Neonatal Intensive Care Unit（NICU）of Jiangsu Province Hospital from June to December 2015. Total RNA was extracted from each blood sample and reverse transcribed into complementary DNA（cDNA）. We used real-time reverse transcription polymerase chain reaction（RT-PCR）technique to determine the serum miR-21 expression levels in the RDS and the non-RDS groups（n=30 and n=21，respectively）. Unpaired student t-test was used to compare the significance between groups. Results：The serum miR-21 expression level in the RDS group was significantly higher than that in the control group（P < 0.05）. The miR-21 expression levels in the moderate to severe RDS groups appeared to be higher than that in the mild groups，however，statistic significance was not reached in our study. In addition，the RDS group that received pulmonary surfactant therapy had significant increase in miR-21 expression levels（P < 0.05）. Conclusion：Serum miR-21 expression level is closely related to the development of infant RDS，suggesting that miR-21 is involved in the pathogenesis of infant RDS.

Abstract:Objective：To evaluate the antiplatelet efficacy of ticagrelor in patients after off-pump coronary artery bypass graft（OPCABG）surgery and observe its clinical outcomes. Methods：A total of 90 patients undergoing OPCABG successfully in the First Affiliated Hospital of NMU during May 2015 to June 2016 were enrolled in this study. Patients were randomly divided into three groups. In the first day after surgery，group A was given clopidogrel（75 mg，1/day，oral）；group B was given ticagrelor（90 mg，2/day，oral）；group C was given half amount ticagrelor（45 mg，2/day，oral）. The three groups took aspirin standard dose（100 mg，1/day）at the same time. In 5 to 7 days after OPCABG，platelet inhibition（PI）rate induced by arachidonic acid（AA）and adenosine diphosphate（ADP）（PIADP and PIAA）by TEG was monitored to compare antiplatelet efficacy of the three groups. Major adverse cardiac events（MACE），bleeding events and related adverse reactions were also compared between the groups to evaluate the clinical effect. Results：Compared with group A，PIADP（%）of group B and group C were significantly increased（83.8 ± 17.6 vs. 60.5 ± 25.6，P=0.001；77.9 ± 24.6 vs. 60.5 ± 25.6，P=0.013，respectively）. There was no obvious difference of PIADP（%）between group B and group C（77.9 ± 24.6 vs. 83.8 ± 17.6，P > 0.05）. The three groups were not seen obvious MACE. Compared with group A，incidence of bleeding events in group B was increased，but there was no statistically significant difference（23.3% vs. 16.6%，P > 0.05）. Conclusion：Ticagrelor treatment as antiplatelet therapy in patients after OPCABG was effective and safe，and its antiplatelet efficacy was superior to clopidogrel.

Abstract:Objective：A localization system for multiple magnetic targets based on magnetic sensor rotation detection was studied，which was aimed at realizing human multiple capsule endoscopy positioning and nano-magnetic drug testing. Methods：A three-axis tunneling magnetoresistive sensor was used to make a circular motion near the magnetic source. Three sets of magnetic signals were acquired and transferred by three-dimensional coordinates. The position of the permanent magnet was solved using a positioning algorithm. Results：Through the preparation of LabVIEW PC program，the trajectory of two magnetic target tracking and motion curve storage and playback were achieved. Conclusion：The detection system has the advantages of good real-time，high precision and low cost，it has the potential application value for the positioning of multiple capsules in the human body and the detection of nano-magnetic drugs.

Abstract:Objective：To compare the predictive performance of constructing the early prognostic model of acute paraquat（PQ）poisoning between support vector machine（SVM）and logistic regression. Methods：A total of 152 patients with acute PQ poisoning were collected and the clinical results were observed for 2 months. The patients were divided into two groups with a 3∶2 ratio by the random numerical table method. One group with a total of 91 cases was used as a training sample for selecting variables and establishing predictive models. Another group with a total of 61 cases was used as a validation sample to evaluate the predictive effect of the model. SVM and conventional logistic regression was used as the modeling method. Results：The prediction accuracy of the kernel，polynomial，sigmoid kernel and radial basis function nuclear SVM model was 77.92%，74.03%，75.32% and 79.22% respectively，when being tested by the validation group. The results of performance comparison showed that SVM models performed better than logistic regression model；RBF-SVM was the best among all the models with a sensitivity of 87.5% and a specificity of 70.6%. Conclusion：SVM model could preferably integrate all kinds of prognostic information of PQ poisoning patients，and the established model had better prediction ability，providing a new method for predicting the prognosis of patients with PQ poisoning.

Abstract:The detection rate and incidence of thyroid nodules have increased year by year. Ultrasound-guided fine needle aspiration cytology（FNA）is the gold standard for preoperative evaluation of the nature of thyroid nodules. However，approximately 20% to 30% of thyroid nodules by FNA are not clearly benign or malignant. Patients with such nodules are often received diagnostic surgery. Most nodules are confirmed benign lesions by postoperative pathological findings. Thus，those nodules have accepted unnecessary surgery. Preoperative accurate diagnoses like genetic tests are used to identify benign or malignant nodules to avoid overtreatment. This article reviews the genetic diagnosis advances and clinical value of risk assessment，efficacy prediction，and prognosis for thyroid nodules.