Objective:To screen the differentially expressed genes(DEGs)under high phosphate-induced calcification in the vascular smooth muscle cells(VSMCs)by mRNA high-throughput sequencing technology,and to analyze the key genes and signaling pathways involved in the VSMCs calcification.Methods:The human VSMCs were divided into control group and model group.The cells in model group was exposed to the high-phosphate medium,while the cells in control group were cultured in DMEM supplemented with 10%fetal bovine serum under the same conditions.The VSMCs in two groups,stably transfected,were cultured for 12 d.The morphology of the cells in two groups were observed and photographed under inverted microscope.The DEGs were selected by Hisat2 software,and Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed by Stringtie software from three aspects,such as biological processes(BP),molecular functions(MF),and cellular components(CC).The calcification of the cells in two groups was observed by Von Kossa staining method.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to analyze the expression levels of alkaline phosphatase(ALP),bone morphogenetic protein 2(BMP2),alpha-smooth muscle actin(α-SMA),tumor protein 53(Tp53),glutathione peroxidase 4(GPX4),ferritin light chain 1(Ftl1),and glycosylphosphatidylinositol-specific phospholipase D1(GPLD1)mRNA in the cells in two groups.Results:Compared with control group,there were 2 524 DEGs in the cells in model group,and there were 1 368 upregulated DEGs and 1 156 downregulated DEGs.Clustering of DEGs between the cells in two groups was distinct.The GO functional and KEGG pathway enrichment analysis results showed that the upregulated DEGs were primarily involved in regulating the microtubule cytoskeleton,cell polarity,protein localization,and cell cycle regulation among BPs;in constructing cell membrane,microtubule organization,chromosomes,and kinetochore among CCs;and functioning in phosphatidylinositol phosphate,Rho GTPase protein binding,transmembrane transport,and protein kinase regulatory activity among MFs.Downregulated DEGs were mainly involved in cytoplasmic translation,protein membrane localization,mRNA metabolism,and protein endoplasmic reticulum localization among BPs;in forming ribosome subunits,cell membrane,and autophagy among CCs;and functioning in single-stranded DNA,ribonucleoprotein complex,growth factor binding,regulating protein kinase activity,and catalytic activity among MFs.Seven signaling pathways were significantly enriched in upregulated genes,most notably in the biosynthesis of glycosylphosphatidylinositol(GPI)anchors;whereas 18 signaling pathways were significantly enriched in the downregulated genes,most notably in ferroptosis.The RT-qPCR results showed that compared with control group,the expression levels of GPX4,Ftl1,and Tp53 mRNA in the cells in model group were significantly decreased(P<0.01),while the expression level of GPLD1 mRNA was significantly increased(P<0.01);compared with control group,the expression level of α-SMA mRNA in the cells in model group was significantly decreased(P<0.01),and the expression levels of ALP and BMP2 mRNA were significantly increased(P<0.01).Conclusion:The VSMCs underwent calcification and normal cells exhibit the DEGs.The key signaling pathways in the calcification induced by high phosphate in the VSMCs include ferroptosis and GPI anchor biosynthesis,mediated primarily through GPX4,Ftl1,Tp53,and GPLD1.

Objective:To identify the differentially expressed long-chain non-coding RNA(lncRNA) and mRNA using ribonucleic acid sequencing(RNA-seq), and construct a competing endogenous RNA(ceRNA) regulatory network in mice with sepsis-associated encephalopathy.Methods:Ten clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups( n=5 each) using a random number table method: sham operation group(group Sham) and sepsis group(group Sepsis). Sepsis was induced by cecal ligation and puncture(CLP) in group Sepsis, while group Sham only underwent laparotomy without CLP. Morris water maze test and contextual fear conditioning test were performed to detect the cognitive function on 1 day before CLP and 3 days after CLP. Three mice were randomly sacrificed in group Sham, and 3 mice with the worst results in the cognitive function test were sacrificed in group Sepsis. The hippocampal tissues were obtained for RNA-seq via the BGISEQ-500 platform, and the differentially expressed mRNA and lncRNA were identified. The differentially expressed mRNAs and lncRNAs were visualized and analyzed by Dr. Tom platform provided by Shenzhen BGI Technology Service Co., Ltd., and the ceRNA regulatory network was constructed using the online visualization tool Cytoscape software. Results:Compared with group Sham, the escape latency was significantly prolonged, and the percentage of time of staying at the target quadrants and percentage of time spent freezing were decreased in group Sepsis( P<0.05). A total of 62 differentially expressed lncRNAs were obtained from RNA-seq, of which the expression of 45 lncRNAs was up-regulated and the expression of 17 lncRNAs was down-regulated.There were 282 differentially expressed mRNAs identified from RNA-seq, of which the expression of 173 mRNAs was up-regulated, and the expression of 109 mRNAs was down-regulated.Gene Ontology enrichment analysis revealed that the differentially expressed mRNAs were involved in biological processes such as memory, learning or memory, inflammatory responses, regulation of aging-related behavioral decline, and regulation of synaptic plasticity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that differentially expressed mRNAs were enriched in IL-17 signaling pathway, TNF signaling pathway, NF-κB signaling pathway and etc. KDA analysis was performed on the differentially expressed mRNAs to identify the key driver genes, and the results showed that Ch25h, Il6ra, Lcn2, Sgk1, Nr4a3, Osm, Saa3, Ccl7, Sqle, Dhcr24 were the key SAE genes.A competing endogenous RNA regulatory network was successfully constructed based on 9 lncRNAs, 28 mRNAs and 134 miRNAs in the hippocampus of mice with SAE. Conclusions:The results of RNA-seq find that 10 mRNAs including Ch25h, Il6ra, Lcn2, Sgk1, Nr4a3, Osm, Saa3, Ccl7, Sqle, Dhcr24 and lncRNAs such as Rian, Gm35874 and Gm34347 are key genes regulating SAE in mice. Meanwhile, a ceRNA regulatory network based on lncRNA-miRNA-mRNA is successfully constructed in the hippocampus of mice with SAE.

Objective:To review our clinical experience on total pancreatectomy in treatment of pancreatic diseases.Methods:The clinical data of 25 patients with pancreatic diseases who underwent total pancreatectomy at Nanjing Drum Tower Hospital from February 2016 to October 2019 were retrospectively analyzed. The data on general information, surgical methods, postoperative complications, changes in postoperative pancreatic endocrine and exocrine functions and quality of life were analyzed.Results:There were 16 males and 9 females, aged (60.9±9.4) years. Planned selective surgery was carried out in 16 patients while unplanned operations in 9 patients. There were 15 patients with pancreatic neoplasms with 4 patients who received neoadjuvant or conversion therapy, 6 patients with intraductal papillary mucinous neoplasms, 3 patients with chronic pancreatitis and 1 patient with pancreatic neuroendocrine carcinoma. The total postoperative complication rate was 36% (9/25). One patient died in the perioperative period, and one patient underwent a second operation. After 6 months of operation, 2 of 24 patients had died. The remaining patients had a fasting blood sugar of (8.9±1.6) mmol/L, with an insulin dosage of (30.7±10.6) U/d. The average dosage of trypsin was (1.1±0.3) g/d. The quality of life score reached or exceeded the preoperative levels.Conclusions:Total pancreatectomy was safe and feasible for some pancreatic diseases. For patients with pancreatic cancer, its long-term oncological outcomes need further studies.

OBJECTIVE：To provide refere nce f or the quality control and evaluation of rice-wine processed Coptidis Rhizoma decoction piece. METHODS ：Taking 17 batches of rice-wine processed Coptidis Rhizoma decoction piece from different manufacturers as samples ，HPLC method was adopted to determine the contents of 4 kinds of alkaloids as epiberberine ，coptisine， palmatine and berberine. The compound weights of epiberberine ，berberine，palmatine and berberine were calculated by the subjective and objective combination weighting method （AHP combined with variation coefficient ）. Then the quality evaluation method was used to evaluate the quality of decoction pieces combined with the appearance of decoction pieces and the contents of 4 alkaloids. The percent mass constant was calculated and the grade of rice-wine processed Coptidis Rhizoma decoction piece was classified. RESULTS ：According to the results of content determination of 4 kinds of alkaloids ，among 17 batches of samples ，a total of 13 batches of samples met the requirements of 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. Mass constants of 13 batches of qualified samples were 10.03-26.96，and the percent mass constants were 37.20％-100％. If the percent mass constant ≥ 80％ of the sample was listed as the first-class product ，the sample with the percent mass constant between 50％-＜80％ was classified as the second-class product ，and the rest was listed as the third-class product ，therefore the quality constant of first-class product was ≥21.57，that of second-class product was 13.48-＜21.57，and that of third-class product was ＜13.48. According to the grading standard ，3 batches of 13 batches of qualified samples are classified as first-class products ，6 batches are classified as second-class products ，4 batches are classified as third-class products. CONCLUSIONS ：The established subjective and objective combination weighting method and quality constant method can more scientifically and reasonably classify rice-wine processed Coptidis Rhizoma decoction piece.

Objective To analyse the value of serum alpha fetoprotein(AFP),keratin antigen 21-1(CY-FRA21-1)and neuron specific enolase(NSE)in the diagnosis and prognosis of lung cancer.Methods A total of 89 patients with primary lung cancer were selected as the primary lung cancer group from June 2014 to June 2016.The other 59 cases of benign lung disease were selected as the benign lung disease group,and 63 cases of healthy physical examination were collected as normal group.The content of AFP,CYFRA21-1 and NSE were detected by electrochemiluminescence assay.Results Before treatment,the levels of AFP,CYFRA21-1 and NSE in the benign lung disease group and the primary lung cancer group were significantly higher than those in the normal group,those in the primary lung cancer group were significantly higher than those in the benign lung disease group(P<0.05).The levels of AFP,CYFRA21-1 and NSE in patients with stage Ⅲ aand Ⅲ b-Ⅳ were significantly higher than those in patients with stage Ⅰ - Ⅱ,and those in Ⅲ b- Ⅳ stage were signifi-cantly higher than those in Ⅲ astage(P<0.05).The sensitivity,specificity,accuracy of two joint tests were significantly higher than those of the single test,those of the three joint tests were significantly higher than those of the single test and the two combined tests,and the differences were statistically significant(P<0.05).The AFP,CYFRA21-1 and NSE of the patients in the ineffective group were significantly higher than those in the effective group,and the differences were statistically significant(P<0.05).Conclusion Tumor markers AFP,YFRA21-1 and NSE are of great value in the diagnosis and treatment of lung cancer,joint detec-tion could improve the positive rate and accuracy of diagnosis.

Objective To translate and revise the Atrial Fibrillation-Quality of Life -18 (AF-QoL-18),and to test its reliability and validity. Methods The Chinese version of AF-QoL-18 was developed through the process of translation, back- translation, cultural adaptation, and preliminary experiment. A total of 187 atrial fibrillation (AF) patients in eight hospitals in Jiangsu province were investigated using the Chinese version to test the reliability and validity. Results Exploratory factor analysis identified three factors, including physiological, psychological and sexual dimensions, which could explain 65.055%of the total variance. The content validity index was 0.969. The Cronbach α coefficient was 0.915 and the retest reliability was 0.948 for the total scale. Conclusions The Chinese version AF-QoL-18 has proved to be reliable and valid.It can be used to measure the quality of life of AF patients in China.

Objective To explore the functional improvement of dancers' sensorimotor system after years of dance training.Methods Twenty-three dancers(the dance group) of more than 6 years of dancing experience and twenty-one college students (the control group) without any dance experience were recruited in this study.Then the resting state functional magnetic resonance imaging(RS-fMRI) was conducted for both groups and the data were collected.Regional homogeneity (ReHo),fractional amplitude of low frequency fluctuation (fALFF) and functional connectivity (FC) were compared between the two groups.Results Compared to the control group,significant increase was observed in the ReHo of the bilateral postcentral gyruses,the left superior temporal gyrus,the right precentral gyrus and the middle occipital gyrus and fALFF in the bilateral precentral gyruses of the dance group.Moreover,the connection between the right precentral gyrus and left precentral gyrus and the FC between the right precentral gyrus and bilateral postcentral gyrus enhanced significantly in the dance group compared to the control group.Conclusions The functional modulation of dancers' sensorimotor system may be associated with the long-term dancing experience,which may lead to the changes in action perception,memory processing,motor learning and movement control to enhance the formation of the mode to observe,implement and adjust complex actions.Our study has provided a supporting evidence for dancing-induced brain plasticity.

Objective To explore the prognostic factors of portal hypertension treated with devascularization.Methods A total of 397 patients with portal hypertension underwent devascularization in Nanjing Drum Tower Hospital from February 1993 to April 2014,among which there were 242 male and155 female patients with median age of 48 years.The perioperative data were retrospectively collected.Logistic regression was used to find the risk factors which affect the operative complications.Follow-up evaluation was in progress regularly.Kaplan-Meier survival curve,Log-rank test and Cox regression model were used to find out factors which affect the long-term results.Results All together 397 patients underwent devascularization,in whom 8 patients died perioperative,389 patients discharged successfully.Logistic regression showed that age (≥ 48 years) (X2 =4.559,OR =2.048,P =0.033),red color sign before surgery (x2 =4.959,OR =2.129,P =0.026) and without portosystemic collateral vessels reserved (x2 =13.348,OR =5.122,P =0.000) were risk factors of perioperative complications.The follow-up time was (5.7±4.6) years.Totally 27 patients were lost from follow-up,103 patients died for the disease during follow-up.The survival rate at 1-,3-,5-,10-,15-and 20-years was 93.6％,86.9％,80.1％,59.3％,54.1％ and 38.5％ respectively.Univariate analysis showed that gender (male),age (≥ 48 years),hemorrhage before surgery (≥500 ml per time),hepatitis virus and without portosystemic collateral vessels reserved were risk factors of the long-term survival (P ＜ 0.05).Cox regression analysis showed that age (≥48 years) (X2 =9.850,RR =1.904,P =0.002),hemorrhage before surgery (≥ 500 ml per time)(X2 =34.402,RR =3.273,P =0.000),hepatitis virus (X2 =7.573,RR =2.525,P =0.006) and without portosystemic collateral vessels reserved (x2 =5.905,RR =１.889,P =0.015) were independent risk factors that affect the long-term survival.Conclusion Devascularization with portosystemic collateral vessels reserved has favorable perioperative and long-term outcome,and it definitely is a very safe and effective technique for portal hypertension.

Objective To explore the prognostic factors of portal hypertension treated with devascularization.Methods A total of 397 patients with portal hypertension underwent devascularization in Nanjing Drum Tower Hospital from February 1993 to April 2014,among which there were 242 male and155 female patients with median age of 48 years.The perioperative data were retrospectively collected.Logistic regression was used to find the risk factors which affect the operative complications.Follow-up evaluation was in progress regularly.Kaplan-Meier survival curve,Log-rank test and Cox regression model were used to find out factors which affect the long-term results.Results All together 397 patients underwent devascularization,in whom 8 patients died perioperative,389 patients discharged successfully.Logistic regression showed that age (≥ 48 years) (X2 =4.559,OR =2.048,P =0.033),red color sign before surgery (x2 =4.959,OR =2.129,P =0.026) and without portosystemic collateral vessels reserved (x2 =13.348,OR =5.122,P =0.000) were risk factors of perioperative complications.The follow-up time was (5.7±4.6) years.Totally 27 patients were lost from follow-up,103 patients died for the disease during follow-up.The survival rate at 1-,3-,5-,10-,15-and 20-years was 93.6％,86.9％,80.1％,59.3％,54.1％ and 38.5％ respectively.Univariate analysis showed that gender (male),age (≥ 48 years),hemorrhage before surgery (≥500 ml per time),hepatitis virus and without portosystemic collateral vessels reserved were risk factors of the long-term survival (P ＜ 0.05).Cox regression analysis showed that age (≥48 years) (X2 =9.850,RR =1.904,P =0.002),hemorrhage before surgery (≥ 500 ml per time)(X2 =34.402,RR =3.273,P =0.000),hepatitis virus (X2 =7.573,RR =2.525,P =0.006) and without portosystemic collateral vessels reserved (x2 =5.905,RR =１.889,P =0.015) were independent risk factors that affect the long-term survival.Conclusion Devascularization with portosystemic collateral vessels reserved has favorable perioperative and long-term outcome,and it definitely is a very safe and effective technique for portal hypertension.

Objective To investigate the diagnostic value ofbiomarkers in cerebrospinal fluid in differentiating meningeal carcinomatosis (MC) and tubercular meningitis (TBM).Methods The clinical features and cerebrospinal fluid (CSF) test results of 34 patients with MC and 44 patients with TBM,admitted to our hospital from January 1994 to August 2014,were retrospectively analyzed and compared.Results The main clinical manifestations of MC included headache and bowel and bladder dysfunction,and those of TBM were fever and headache.The cell population of MC and TBM patients increased and the number of leucocyte of TBM patients was significantly larger than that of MC patients (P＜0.05);the CSF of all the patients presented with decreased levels of glucose and chloride,and increased protein level,and higher protein in MC patients was noted than that in TBM patients (P＜0.05).The biomarkers in CSF of MC and TBM patients increased to varying degrees,and the biomarkers,including carcinoembryonic antigen (CEA),cancer antigen (CA) 125,CA 199,CA 153 and CYFRA21-1,in MC group were significantly higher than those in TBM group (P＜0.05).The neuron-specific enolase level in the TBM patients of Ⅲ grade was significantly higher than that in the MC patients (P＜ 0.05).Conclusion The detections of tumor biomarkers in cerebrospinal fluid is a tool in differentiating MC and all stages of TBM,especially with the combined detections of CEA,CA125,CA199 and CYFRA21-1.