ObjectiveTo identify the pharmacodynamic material basis of Ruyi Zhenbaowan in relieving neuropathic pain by integrating the calculation of biological network proximity and pharmacokinetic characterization. MethodThe interaction network of "drug candidate target-related gene of disease" was constructed by Cytoscape 3.8.2， and the average shortest path value of each drug putative target acting on neuropathic pain-related genes in this network was calculated by Pesca 3.8.0 tool so as to evaluate the network proximity between them， and screen prescription candidate targets with strong intervention efficiency and their corresponding potential effect components. After that， plasma and cerebrospinal fluid samples were collected from rats after administration of Ruyi Zhenbaowan at set time points， and the contents of potential effect components in samples was quantified by ultra performance liquid chromatography-quadrupole-ion trap mass spectrometry（UPLC-Q-TRAP/MS）， and drug concentration-time curves were plotted， then the pharmacokinetic parameters were calculated by DAS 2.1.1. ResultBy evaluating the network proximity between candidate targets and neuropathic pain-related genes in the interaction network， a total of 40 putative targets of Ruyi Zhenbaowan with strong intervention effects on neuropathic pain-related genes， such as estrogen receptor 1（ESR1）， cyclic adenosine monophosphate（cAMP）-dependent protein kinase catalytic subunit alpha（PRKACA） and protein kinase B1 （Akt1）， and 10 corresponding potential effect components， such as glycyrrhizic acid and betulinic acid， were obtained. Pharmacokinetic characterization showed that among the 10 potential effect components， gallic acid， apigenin-7-O-glucuronide， glycyrrhizic acid and apigenin were well absorbed and metabolized in plasma and cerebrospinal fluid， with long onset time and good bioavailability. ConclusionFrom the perspective of efficacy-target-constituent-pharmacokinetic， this study analyzes the main effective materials of Ruyi Zhenbaowan， such as glycyrrhizic acid， gallic acid， apigenin-7-O-glucuronide and apigenin， which have a high exposure in plasma or cerebrospinal fluid and have a strong intervention effect on neuropathic pain. The related results provide reliable experimental evidences for clarifying the material basis and developing quality standards of Ruyi Zhenbaowan.

Objective:To explore the differences of immune reconstitution between peripheral blood stem cell transplantation and umbilical cord blood transplantation.Methods:A total of 300 patients (aged 18 (8, 33), 163 males and 137 females) with malignant hematological diseases who received allogeneic hematopoietic stem cell transplantation in the First Affiliated Hospital of University of Science and Technology of China from January 2018 to March 2020 were enrolled in this study, including 255 cases of umbilical cord blood transplantation and 45 cases of peripheral blood stem cell transplantation. Multicolor flow cytometry was applied to analyze lymphocyte subsets of the percentages and absolute counts in the two donor types and peripheral blood of patients after receiving hematopoietic stem cell transplantation. The differences between the two grafts were compared, and the lymphocyte subsets results were evaluated at 1, 2, 3, 4, 6, 9, 12, and 18 months after transplantation. 18-month disease-free survival (DFS) within the 300 patients under the two transplantation types were retrospectively analyzed.Results:1. The proportion of NKT cells in peripheral blood group was significantly higher than that in cord blood group (2.79% vs 0.24%, P<0.001). 2. The proportion of helper T cells in the UCBT group was higher than that in the PBSCT group, as well as the counts 6 months after transplantation ( P<0.05). 3. The proportion of NK1 cells (3 rd to 9 th month) and count (4 th to 12 th month) in UCBT group were significantly higher than those in PBSCT group ( P<0.05). 4. NKT cells in the UCBT group were lower than those in the PBSCT group (proportion and count) throughout the monitoring process ( P≤0.001). 5. The proportion of DNT cells (within 1 year) and count (within 6 months) in the UCBT group were significantly lower than those in the PBSCT group ( P<0.05). Conclusions:Compared with the peripheral blood stem cell transplantation group, the umbilical cord blood transplantation patients had a faster rate of lymphocyte reconstitution, and patients received umbilical cord blood transplantation had a stronger ability of immune reconstitution and could achieve long-term hematopoiesis.

Objective:To study the changes of urinary metabolic profile, screen metabolic ions characterization with clinical diagnostic value, and a disease differentiation model establishment, in an attempt to help the clinical diagnosis of hepatocellular carcinoma patients.Methods:A case-control study was conducted. Ultra-performance liquid chromatography/mass spectrometry (UPLC-MS) were used to analyze urine samples of 32 patients with hepatocellular carcinoma, 28 patients with liver cirrhosis and 28 healthy persons, respectively. The orthogonal partial least squares discriminant analysis (OPLS-DA) and the principal component analysis (PCA) model were constructed using MZmine2.0 and SIMCA-P + 12.0.1.0 software for preliminary screening of metabolites. The metabolic ions selected in the final test were analyzed by SPSS, and the markers were analyzed and screened by one-way analysis of variance. Finally, the sensitivity and specificity of the selected markers were analyzed by calculating the area under the receiver operating characteristic (ROC) curve. One-way analysis of variance was used to compare quantitative indicators between groups.Results:OPLS-DA model parameters were R2X = 35.3%, R2Y = 86.9%, and Q2 = 72.2%, which had a good identification value. A total of 26 characteristic ions were screened, of which 17 were identified. 14, 19-Dihydroaspidospermatine had a high value in distinguishing healthy person with hepatocellular carcinoma patients, and the area under the receiver operating characteristic curve was higher than 0.9. The area under the ROC curve for distinguishing liver cancer with liver cirrhosis patients was 0.88, which was higher than the ROC curve of alpha-fetoprotein (0.75).Conclusion:Based on the UPLC-MS platform, the PCA and OPLS-DA models were successfully constructed, and the characteristic metabolic ions in the urine were extracted and identified, which has a certain value in assisting clinical screening of hepatocellular carcinoma patients.

Curriculum integration and problem based learning (PBL) is the development trend of medical education all over the world,and it is an effective mode to train high-quality medical talents to meet the needs of the times and national medical and health services.However,PBL is difficult to popularize in an all-round way at present due to the large enrollment of clinical medicine major in medical colleges and universities in China.Aiming at integrating knowledge and focusing on ability,Nanjing Medical University implemented a problem-based two-way integrated case teaching after top-level design,careful trial and steady promotion and has achieved initial results and improved the clinical competency of medical students.

Objective To explore the expression feature of long non-coding RNA (lncRNA) 1010001N08Rik in hyperoxia-induced bronchopulmonary dysplasia (BPD) and predict the mechanism that 1010001N08Rik might be involved in the occurrence and development of BPD by a series of bioinformatics analysis.Method The sequence,genomic position and structure characteristics of 1010001N08Rik were acquired from UCSC genome browser,and its target gene was predicted by Ensemble database.We successfully established the animal model of BPD by making newborn C57BL/6J mice exposed to 95％ concentrations of ambient oxygen for seven days.The expression of 1010001N08Rik and Gata 6 were detected using real-time quantitative polymerase chain reaction (PCR).Student's t test was used to compare their expression levels during the BPD process.Result The relative expression of 1010001N08Rik in BPD process at d1,d3,d5,d7 was 1.21 ± 0.33,2.02 ± 0.41,2.95 ± 0.45,4.20-± 0.48 respectively,and there were significant difference between adjacent time points (P ＜ 0.05).The relative expression of Gata 6 mRNA was 0.92 ±0.30,1.10 ± 0.31,0.86 ± 0.24,0.45-± 0.08 respectively,and there was significant difference between d5 and d7 (P ＜0.05).1010001N08Rik had highly conserved property among different species.The chromosomal regions of 1010001N08Rik existed transcriptional factors binding locations and epigenetic regulation clues,and its possible candidate target gene was Gata 6.Conclusion The expression of 1010001N08Rik increased during the formation process of BPD.Bioinformatics analysis and preliminary experiment results suggested that 1010001N08Rik might participate in the process of BPD by down-regulating Gata 6 expression.

Objective: To determine the diagnostic value of serum cystatin C (Cys C) for acute kidney injury (AKI) in patients with liver cirrhosis. Methods: Serum Cys C levels in 150 liver cirrhosis patients (88 AKI and 62 non-AKI patients) were measured by the Particle-Enhanced Nephelometric Immuno-Assay. The accuracy of serum Cys C for the diagnosis of AKI in liver cirrhosis was evaluated by the ROC curve. Results: Liver cirrhosis patients with AKI had significantly higher serum Cys C levels [2.37 (1.75-2.83) mg/L] than those without AKI [0.97 (0.85-1.09) g/L] (P <0.001). Serum Cys C level was highest in the acute tubular necrosis group [5.41 (2.77-6.19) mg/L], followed by the hepatorenal syndrome group [2.55 (2.28-3.59) mg/L] and prerenal azotemia group [2.07 (1.70-2.41) mg/L], and the serum Cys C level was significantly different between the three groups (P <0.001). In addition, patients with AKI were further divided into infection group and non-infection group. Serum Cys C level was significantly higher in the infection group than in the non-infection group (P <0.05). The area under the ROC curve of serum Cys C for the diagnosis of AKI in liver cirrhosis was 0.99 (0.98-1.00) at a cut-off value of 1.36 mg/L, and the sensitivity and specificity were 97% and 95%, respectively. Conclusion Serum Cys C is a good marker for detecting AKI in liver cirrhosis, and the different levels of increase in Cys C may be useful in differentiating the different types of AKI.

Objectives This research explored the characteristics of changes in the serum metabolic profile of preeclampsia pregnancy(PE) to establish the disease distinguish model and screen characteristic metabolic markers with potential diagnostic value for preeclampsia.Methods From August 2014 to January 2016,samples in three groups were collected at Tianjin Third Central Hospital.Thirty-one clinically diagnosis patients with preeclampsia,25 normal pregnancy women and 29 healthy volunteers of childbearing age were enrolled.Ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze serum metabolites of PE group (31 patients with preeclampsia),P group (25 normal pregnancy women) and Normal group (29 healthy volunteers of childbearing age).Nonparametric test analyzes were used to analyze the data and find the specific metabolites.Results This research established the principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) disease distinguish model for PE group,P group and Normal group.To distinguish PE group,P group and Normal group,15 characteristic metabolites were identified.Eight kinds of glycerol phospholipid (including 7 kinds of hemolysis phosphatidyl choline and 1 kind of lysophospholipids acid) and 1 kind of sphingomyelin in PE group were higher than that of normal pregnancy group.The difference had statistically significant(Z of the metabolites were 2.32,3.34,3.21,2.60,2.22,3.40,3.58,5.84,2.70 respectively,all P<0.05).1,25-Dihydroxyvitamin D3-26,23-lactone and 24-Oxo-1alpha,23,25-trihydroxyvitamin D3 in PE group were higher than that of P group and Normal group,which had a statistics difference (Z of the metabolites were 2.01,3.89,3.26,2.34 respectively,all P<0.05).Conclusions Metabolomics distinguish model has a good ability to distinguish PE group,P group and Normal group.Serum characteristic metabolites can successfully reflect the status of fat,calcium and phosphorus metabolism of preeclampsia patients and provide high value for prediction,diagnosis and treatment.

AIM:Atherosclerotic calcification is highly linked with plaque instability and cardiovascular events .Adenosine monophosphate-activated protein kinase ( AMPK) has been involved in the pathogenesis of various cardiovascular disease .The contributions of AMPKαsubunits to the development of atherosclerotic calcification in vivo remained unknown .We hypothesized that AMPKαsubunits may play a role in the development of atherosclerotic calcification .METHODS: Atherosclerotic calcification was generated by 24-week fed of western diet in ApoE-/-background mice .Calcification was evaluated in aortic roots and innominate arteries of ApoE-/-mice or in mice with dual deficiencies of ApoE and AMPKαsubunits globally ( AMPKα1 and AMPKα2 ) , or vascular smooth muscle cell ( VSMC)-specific or macrophage-specific knockout of AMPKα1 with atherosclerotic calcification pone diet . The mechanism of AMPKα1 in regulating Runx2 was further explored in human aortic VSMC .RESULTS: Ablation of AMPKα1 but not AMPKα2 in ApoE-/-background promoted atherosclerotic calcification with increased Runt -related transcription factor ( Runx2 ) expression in VSMC compared with ApoE-/-mice.Conversely, chronic administration of metformin, which activated AMPK, markedly reduced ath-erosclerotic calcification and Runx2 expression in ApoE-/-mice but had less effects in ApoE-/-/AMPKα1 -/-mice.Furthermore, VSMC-but not macrophage-specific deficiency of AMPKα1 in ApoE-/-background promoted atherosclerotic calcification in vivo com-pared with the controls .AMPKα1 silencing in human aortic VSMC prevented Runx 2 from proteasome degradation to trigger osteoblastic differentiation of VSMC .Conversely , activation of AMPK led to Runx 2 instability by inducing its small ubiquitin-like modifier modifi-cation (SUMOylation).Protein inhibitor of activated STAT-1 (PIAS1), the SUMO E3-ligase of Runx2, was directly phosphorylated by
AMPKα1 at serine 510, to enhance its SUMO E3-ligase activity.Ablation of PIAS1 serine 510 phosphorylation inhibited metformin-in-duced Runx2 SUMOylation, and subsequently prevented the effect of metformin on reducing oxLDL-triggered Runx2 expression in hu-man aortic VSMC.CONCLUSION:Deficiency of AMPKα1 in VSMC increases Runx2 expression and promotes atherosclerotic calcifi-cation in vivo.AMPKα1 phosphorylates PIAS1 to enhance Runx2 SUMOyalation and subsequent degradation .

Objective To assess the efficacy and safety of lamotrigine for absence seizures in children and adolescents . Methods Databases of PubMed ,the Cochrane Library ,EMbase ,CENTRAL ,VIP ,WanFang ,CBM and CNKI were electron-ically searched till August ,2014 for clinical trials on lamotrigine for absence seizures in children and adolescents .All literature were screened by two reviewers independently according to the inclusion and exclusion criteria .The data was extracted ,and the methodological quality was assessed .Then ,meta-analysis was performed using RevMan 5 .2 .Results Seven trials were in-cluded involving a total of 721 patients .The results of methodological qualities were two studies rated as A-class ,three studies rated as B-class and two studies rated as C-class .Meta-analysis results showed that the efficacy of lamotrigine monotherapy for absence seizure in children and adolescents was better than placebo ,but efficacy of lamotrigine was lower than valproic acid and ethosuximide .The adverse reaction rates of lamotrigine were with no significant difference compared with valproic acid and et-hosuximide .Conclusion Lamotrigine monotherapy was effective for absence seizures in children and adolescents and was well tolerated .Lamotrigine was a good choice for patients that are intolerable to valproic acid or ethosuximide .

Objective To investigate the value in the diagnosis of tumor of the stomach by hypotonic water filling method com-bined with CT multi planar reconstruction (MPR).Methods CT image data of 21 5 cases with gastric tumor confirmed by operation and pathology in our hospital were analysed retrospectively.Conventional CT enhanced scan was obtained in patients with the stom-ach hypotonic water filling condition,and MPR CT characteristics of lesions were observed.Results In the 21 5 cases of gastric be-nign or malignant lesions,MPR showed 5 pathological types in 210 cases.In the conventional CT examination,the tumor diagnosis rate had obvious improvement in different gastric parts and types of the stomach tumors through CT MPR.Conclusion There is high detection rate in the diagnosis of gastric tumors using hypotonic water filling method with MPR,which can accurately display invasion and metastasis,and reduce the misdiagnosis and missed diagnosis in gastric tumor.