Abstract:Considering the difficulties and challenges faced by the medical and health services in China, as well as the features and advantages of traditional Chinese medicine (TCM), the State Council Information Office of the People's Republic of China issued White Paper-Traditional Chinese Medicine in China, and the 25^th Meeting of the Standing Committee of the 12^th session of the National People's Congress of the People's Republic of China approved the Law of the People's Republic of China on Traditional Chinese Medicine. The White Paper and the Law of the People's Republic of China on Traditional Chinese Medicine have further established the position and role of the TCM in the medical and health services in China. This paper introduced the challenges for the medical and health services, the characteristics of TCM theory, and described the equal importance of TCM and western medicine in the maintaining and improving people's health.

Abstract:Objective To determine whether the cytokeratin 19 (CK19) positive (CK19⁺) cells in mouse liver have the potential of differentiation into mature hepatocytes. Methods The CK19^CreERT/Rosa26-GFP double-transgenic mice were obtained by hybridizing CK19^CreERT mouse with Rosa26-GFP mouse and the GFP-labeled CK19⁺ cells were observed in mouse liver after tamoxifen (TM) injection. Then we constructed the 3, 5-diethoxycarbonyl-1, 4-dihydrocollidine (DDC)-induced liver injury mouse model and carbon tetrachloride (CCl₄)-induced liver injury mouse model in CK19^CreERT/Rosa26-GFP double-transgenic mouse and determined the differentiation potential of GFP-labeled CK19⁺ cells by frozen sections and immunofluorescence staining of liver tissues. Results The CK19^CreERT/Rosa26-GFP double-transgenic mice were obtained and the CK19⁺ cells were genetically labeled with GFP after TM injection. In DDC model, we detected GFP⁺ cholangiocytes in proliferating ductules, in which CK19, a marker for biliary epithelial cells was expressed, and the proportion of GFP⁺ cholangiocytes in the DDC model group ([63.5±6.3]%) was significantly higher than that in the control group ([53.6±4.8]%, P<0.05). In CCl₄ model, we identified GFP⁺ hepatocytes in the liver parenchyma; we also found that the hepatocyte-specific marker albumin was expressed in the GFP⁺ hepatocytes; and the proportion of GFP⁺ hepatocytes in the CCl₄ model group ([0.15±0.02]%) was significantly higher than that in control group ([0.008±0.003]%, P<0.01). Conclusion There are liver stem cells with a potential for hepatic differentiation in the CK19⁺ cells in mouse liver, which may provide a new clue for the identification of liver stem cells.

Abstract:Objective To analyze the risk factors of post-operative complications of endoscopic retrograde cholangiopancreatography (ERCP) in patients with liver cirrhosis, and to establish a simple and practical scoring system. Methods We retrospectively analyzed the clinicopathological data of 223 patients with liver cirrhosis who underwent ERCP in Changhai Hospital of Second Military Medical University from Jan. 2009 to Dec. 2015. The univariate analysis and logistic multivariate regression analysis were used to study the effects of pre-operative status, lab parameters and operation performance on post-operative complications. After screening for independent risk factors and transforming the continuous variables into categorical variables, we assigned the scores according to the β value of each independent risk factor, established a new scoring system to predict the occurrence of post-operative omplications, and then drew the receiver operating characteristic (ROC) curve to assess its predicting efficacy for complications. Results Multivariate regression analysis showed that total bilirubin (TBIL) was an independent risk factor for post-operative complications with an OR of 1.003 (95%CI 1.001, 1.005), and albumin (ALB) as a protective factor with an OR of 0.935 (95%CI 0.879, 0.994). The scoring system established according the two factors was as follows:TBIL ≤ 31.4 μmol/L for 0 point, 31.5-102.5 μmol/L for one point, ≥ 102.6 μmol/L for two points; ALB ≥ 31 g/L for 0 point, ≤ 30 g/L for one point. The area under ROC curve (AUC) of the new scoring system was 0.689, which was similar to the Child-Pugh classification (0.700) and the model for end-stage liver disease (MELD) score (0.692) by ROC analysis. All patients were divided into two parts by 1.5 points according to the new scoring system (0-1 point was set as low-risk, and 2-3 points was high-risk). The sensitivity of the new scoring system was 89.4%, specificity was 41.1%, accuracy rate was 49.3%, the positive predictive value was 23.7%, and the negative predictive value was 95.0%. Conclusion The new scoring system for post-ERCP complications is easy-to-use and can be used to identify patients at high-risk.

Abstract:Objective To investigate whether antihypertensive agent verapamil can inhibit hepatitis C virus (HCV) infection via reducing the expression of thioredoxin-interacting protein (TXNIP) in hepatocytes of the host. Methods Human hepatocellular carcinoma Huh7.5.1 cells were treated with different concentrations of verapamil, and then the mRNA and protein expressions of TXNIP were detected by qPCR and Western blotting, respectively. The HCV infection level of Huh7.5.1 cells was determined 48 h after treatment with verapamil and cell culture-derived HCV (HCVcc). We observed the effect of verapamil on the Huh7.5.1 cells with TXNIP silenced by siRNA after infected by HCVcc. Huh7.5.1 cells were transfected with expression plasmid of enhanced green fluorescent protein (EGFP) controlled by TXNIP promoter (pTXNIP-EGFP), and then the effect of verapamil on transcriptional activity TXNIP promoter was analyzed. Results Compared with the control group, verapamil (100, 200, 400 μmol/L) significantly inhibited TXNIP expression and HCV infection in Huh7.5.1 cells in a dose-dependent manner (P<0.05). Furthermore, verapamil reduced EGFP expression in Huh7.5.1 cells transfected with pTXNIP-EGFP in comparison with the control group (P<0.05). Conclusion Verapamil can inhibit HCV infection via reducing TXNIP expression, which may be associated with the inhibition of TXNIP promotor transcriptional activity.

Abstract:Objective To identify the Uighur medicine with good antifungal activity via determining the in vitro antifungal activities of 122 kinds of Uighur medicines. Methods The 75% ethanol extracts of 122 kinds of Uighur medicines were prepared by ultrasonic extraction, and anti-deep fungal medicines were screened according to the M38-A2 and M27-A3 broth microdilution method approved by the Clinical Laboratory Standards Institute (CLSI). The broth microdilution checkerboard method was used to determine the synergistic effect of Uighur medicine and fluconazole against drug-resistant Candida albicans. Tannin content was determined by phosphomolybdotungstic acid-casein method, and the relationship between antifungal activity of different extractions and contents of tannin was studied. Results and conclusion At a determination concentration of 2 000 μg/mL, 21 kinds of Uighur medicines such as Terminalia chebula Retz. and Terminalia bellerica Roxb. had inhibitory effect on Cryptococcus neoformans, 15 kinds of Uighur medicines such as Terminalia chebula Retz. and Terminalia bellerica Roxb. had inhibitory effect on Candida albicans, and Curcuma longa L. had inhibitory effect on Aspergillus fumigatus. Curcuma longa L. had the widest antifungal spectrum and showed good inhibitory effect on deep and superficial fungus. Nine kinds of Uighur medicines showed synergistic effects with fluconazole against Candida albicans. Antifungal activity parts of Terminalia chebula Retz., Terminalia bellerica Roxb., Bergenia purpurascens Engl., Acacia senegal Willd and Rhus coriaria L. were concentrated in ethyl acetate, and the minimum inhibitory concentration of different solvent extraction was directly proportional to the content of tannin.

Abstract:Objective To compare the effects of dexmedetomidine and propofol on sedation and hemodynamics of patients undergoing mechanical ventilation after cardiac valve surgery. Methods Sixty patients who received mechanical ventilation after cardiac valve surgery were randomly divided into dexmedetomidine group (Group D, n=30) and propofol group(Group P, n=30). The patients in group D received the sedation formula of dexmedetomidine 0.2-0.7 μg/(kg·h) and sufentanil 0.15 μg/(kg·h), while those in group P received the sedation formula of propofol 1.2-3.0 mg/(kg·h) and sufentanil 0.15 μg/(kg·h). The dose of sedation was regulated in order to maintain bispectral index (BIS) score at 70-85 and Richmond agitation-sedation scale (RASS) at -2——3. Hemodynamic parameters of the patients were assessed at pre-operation (T₁), 2 h post-cardiopulmonary bypass (CPB, T₂), 4 h post-operation (T₃) and 12 h post-operation (T₄). We compared and analyzed the sedation scores of BIS and RASS, and incidences of complications such as mean arterial pressure (MAP) decrease, bradycardia, delirium or agitation of the patients in two groups. Results There was no significant difference between two groups in BIS score and RASS score (P>0.05). There was a significant difference in hemodynamics change between the two groups. The MAP in Group P was significantly lower than that in Group D at T₂ (P<0.05). The cardiac index in Group D was significantly higher than that in Group P at T₃ (P<0.05) and T₄ (P<0.01). Arterial blood lactate contents in Group D were significantly lower than that in Group P at T₂, T₃ and T₄ (P<0.05, P<0.01).There was no significant difference between two groups in systemic vascular resistance index, stroke volume, right ventricular ejection fraction or oxygen saturation of mixed venous blood (P>0.05). The vasoactive agent dosages were not different between two groups (P>0.05). There was no significant difference between two groups in incidences of post-operative MAP decrease or bradycardia (P>0.05) and the incidences of post-operative delirium or agitation in Group D was significantly lower than those in Group P (P<0.05). Conclusion Sedation effect of both drugs is good in our study. Dexmedetomidine has relatively less influence on hemodynamics, and it can reduce the incidence of post-operative agitation or delirium. Dexmedetomidine seems to be more appropriate for post-operative sedation for the patients undergoing mechanical ventilation after cardiac valve surgery.

Abstract:Objective To explore the clinical efficacy of continuous hyperthermic peritoneal perfusion chemotherapy (CHPPC) in the treatment of advanced cholangiocarcinoma and its long-term impact on survival time. Methods This was a retrospective study of 103 patients with advanced cholangiocarcinoma from Aug. 2014 to Jun. 2016 in Eastern Hepatobiliary Surgery Hospital of Second Military Medical University. The 46 patients in CHPPC group underwent CHPPC and 57 patients in control group received conventional intravenous chemotherapy. We recorded and analyzed the quality of life (KPS score), clinical efficacy, liver function indexes (including aspartate aminotransferase[AST], alanine aminotransferase[ALT], γ-glutamyl transferase[γ-GT], total bilirubin[TB] and alkaline phosphatase[ALP]), serum carbohydrate antigen 19-9 (CA19-9), adverse reaction and survival time of the patients in the two groups. Results The improvement rate of quality of life in the CHPPC group was significantly higher than that in the control group (34.78%[16/46] vs 19.30%[11/57], P<0.05). The effective rate of the CHPPC group was significantly higher than that of the control group (67.39%[31/46] vs 40.35%[23/57], P<0.01). The median levels of AST, ALT, γ-GT, TB, ALP and CA19-9 in the CHPPC group were significantly lower than those in the control group (P<0.05). The incidences of aleucocytosis, thrombocytopenia and liver function injury in the CHPPC group were significantly lower than those in the control group (P<0.05), and there was no significant difference between two groups in the incidence of reduction of hemoglobin, gastrointestinal adverse reaction or renal injury (P>0.05). The survival time of the patients in the CHPPC group was significantly higher than that in the control group ([12.00±2.47] months vs[6.00±0.80] months, P<0.01). Conclusion CHPPC shows a significant clinical efficacy in the treatment of advanced cholangiocarcinoma. It can effectively decrease the median level of serum CA19-9, improve the liver function and quality of life,and prolong the survival time of the patients.

Abstract:Objective To explore the effect of microRNA-205 (miR-205) expression on migration and invasion of human melanoma cell lines (A375 and A2058) and its related molecular mechanism. Methods The miR-205 over-expressed melanoma cells (miR-205 group) were constructed via transfecting A375 and A2058 cell lines with Lenti-miR-205, and the cells transfected with blank vector virus were taken as control group (NC group). The migration and invasion ability of the cells in two groups were detected by scratch assay and Transwell assay, respectively. The confocal laser scanning microscopy was employed to observe the cell morphology and the cell number and fluorescence intensity of the cells expressing E-cadherin protein. Western blotting was performed to detect the protein expressions of E-cadherin, Twist, intergrin, vimentin, Zeb1, matrix metalloproteinase (MMP)-2 and MMP-9. Results At 8 h and 24 h of cells culture, the migration ability of A375 and A2058 cells in miR-205 group was significantly higher than that in NC group (P<0.01); the invasion ability of two kinds of cells in miR-205 group was lower than that in NC group (P<0.01). A375 and A2058 cells over-expressing miR-205 were transformed from shuttle, stroma type to round and epidermal type. Compared with the NC group, the E-cadherin expression of the cells in miR-205 group was significantly increased (P<0.01) and the cell number and fluorescence intensity of the cells expressing E-cadherin was increased, while the protein expressions of Twist, intergrin, vimentin, Zeb1, MMP-2 and MMP-9 were significantly decreased (P<0.01). Conclusion MiR-205 can inhibit the migration and invasion abilities of melanoma cells by inducing E-cadherin expression and reversing epithelial-mesenchymal transition.

Abstract:Objective To explore the protective effect of cannabidiol (CBD) on palmitic acid (PA)-induced hepatocytes injury and to investigate the underlying mechanism associated with autophagic flux. Methods Primary cultured rat hepatocytes were treated with 1 or 5 μmol/L CBD for 24 h and Western blotting was performed to detect autophagy-related protein microtubule-associated protein 1 light chain 3 (LC3) and p62 expressions. Then the hepatocytes were divided into 4 groups and received different interventions, including PA group (the hepatocytes were stimulated with 800 μmol/L PA), PA+CBD group (the hepatocytes were co-stimulated with 800 μmol/L PA and 5 μmol/L CBD), PA+CBD+CQ group (the hepatocytes were co-stimulated with 800 μmol/L PA, 5 μmol/L CBD and 50 nmol/L autophagy inhibitor chloroquine[CQ]) and negative control group (an equal volume of 0.03% DMSO was added to the culture medium); the hepatocytes in all groups were treated for 24 h. We used Western blotting to detect LC3 and p62 proteins expressions, flow cytometry to determine apoptosis, qPCR assay to detect the mRNA expressions of CCAAT/enhancer-binding protein homologous protein (CHOP), glucose-regulated protein 78 (GRP78) and X-box-binding protein 1 (XBP-1), and the Rh123 and lucigenin fluorescent probes to detect the mitochondrial membrane potential and reactive oxygen species (ROS) content, respectively. Results The ratio of LC3-Ⅱ to LC3-Ⅰand p62 protein expression had no change in the cultured rat hepatocytes treated with 1 or 5 μmol/L CBD. Compared with the negative control group, the ratio of LC-Ⅱ to LC3-Ⅰand p62 protein expression were significantly increased (P<0.05), the apoptosis was significantly risen (P<0.05), the mRNA expressions of CHOP, GRP78 and XBP-1 were significantly increased (P<0.05), mitochondrial membrane potential was significantly decreased (P<0.05), and the ROS content in mitochondrial was significantly increased (P<0.05) in the PA group. Compared with PA group, the hepatocytes in the PA+CBD group showed an improved autophagic flux, increased apoptosis, reduced endoplasmic reticulum stress and mitochondrial dysfunction (all P<0.05). The protective effect of CBD on PA-induced hepatocytes injury was significantly inhibited by co-incubation with CQ (P<0.05). Conclusion CBD can attenuate PA-induced hepatocytes injury through promoting autophagic flux, reducing hepatocyte apoptosis, and improving endoplasmic reticulum stress and mitochondrial dysfunction.

Abstract:Objective To explore the effect of metformin on thyroid hormone in rats with hypothyroidism and its mechanisms. Methods A total of 36 three-month-old male SD rats were randomly divided into normal control (NC) group and hypothyroidism (HT) group, metformin (Met) group. The rats in HT group and Met group was fed with 0.01% amino three triazole (AMT) water for 4 weeks to create hypothyroidism rat model, while NC group were fed with normal water. After modeling, the rats in Met group were intragastrically administrated with metformin 200 mg/kg, while the rats in HT group and NC group were intragastrically administrated with equal volume of 0.05% carboxymethylcellulose sodium (CMC-Na). After intragastric administration for eight weeks, the rats in each group were sacrificed to observe thyroid tissues with H-E staining, and to detect prothyrotropin-releasing hormone (proTRH) mRNA expression with qPCR assay. After intragastric administration for 8 and 12 weeks, serum T3 and T4 levels were detected using electrochemiluminescence assay, rat thyroid sodium/iodide symporter (NIS) mRNA and protein expressions were detected by qPCR and Western blotting, respectively. Results T3 and T4 levels of rats in HT group were significantly lower than those in NC group after 8 and 12 weeks of intragastric administration, while T3 and T4 in Met group were significantly higher than those in HT group (P<0.05). At week eight of intragastric administration, H-E staining showed that there was a greater variability in the size of folliculars, in which a large number of colloid and vacuoles were observed in the NC group; thyroid folliculars in the HT group were shrunk, the follicular epithelial tissues were columnar, and vacuoles around the follicles were decreased; and the thyroid follicular epithelial tissues hyperplasia in the Met group was slightly improved versus the HT group. At week eight of intragastric administration, the results of qPCR showed that thyroid tissue NIS mRNA and hypothalamic proTRH mRNA expressions in HT group were higher than those in NC group, while those in Met group were lower than those in HT group (P<0.05); Western blotting results showed that NIS protein expression in Met group was significantly lower than that in HT group (P<0.05), and there was no significant difference in NIS protein expression between HT group and NC group (P>0.05). After 12 weeks, there was no significant difference in NIS mRNA expression between HT group and Met group (P>0.05), while NIS protein in Met group was significantly higher than that in HT group (P<0.05). Conclusion Metformin can increase the thyroid hormone level, but its short-term mechanism may not be related to promoting the level of NIS.

Abstract:Objective To explore the impact of long-term administration of different nucleos(t)ide analogues on estimated glomerular filtration rate (eGFR) in post-liver transplantation patients with chronic hepatitis B virus (HBV) infection. Methods In this retrospective clinical cohort study, we included 177 post-liver transplantation patients treated with entecavir (ETV; ETV group), telbivudine (LDT; LDT group), lamivudine (LAM; LAM group) or adefovir dipivoxil (ADV; ADV group) between Aug. 2008 and Oct. 2014 in our hospital, and clinical data of all patients were analyzed. The eGFR levels in post-operation of 3 (baseline), 9, 15, 21 and 27 months were analyzed by modification of diet in renal disease (MDRD) formula and chronic kidney disease epidemiology collaboration (CKD-EPI) formula. Logistic regression method was used to analyze risk factors for ≥ 20% mean change of baseline eGFR level at 27 months after operation. Results Sixty-six of 177 patients received treatment with ETV, 21 with LDT, 61 with LAM and 29 with ADV. Compared with baseline, the eGFR levels of the patients in LAM group and ADV group were significantly decreased at 9 months after liver transplantation (P<0.05), and there was a decreasing tendency during the 27-month follow-up (P<0.001). However, the eGFR level of the patients in LDT group showed a significant increase at 9 months after post-operation (P<0.05), and there was an increasing tencdency during the 27-month follow-up (P<0.001). There was no significant change in eGFR level during the follow-up period in ETV group. Multivariate logistic regression analysis showed that LAM or ADV treatment were independent risk factors for ≥ 20% decrease in the eGFR level at 27 months after operation compared with baseline (P<0.01), and baseline blood urea nitrogen level and LDT treatment were independent predictive factors for ≥ 20% increase in the eGFR level (P<0.01). Conclusion In post-liver transplantation patients, long-course treatment with LAM or ADV may be a potential cause of nephrotoxicity, while LDT treatment can increase the patient's eGFR level, and ETV has no significant influence on the eGFR.

Abstract:Objective To determine whether serum deprivation can induce transdifferentiation of cardiac fibroblasts into endothelial cells in the ischemic and hypoxic microenvironment of acute myocardial infarction. Methods Fibroblasts were separated and cultured from hearts of new born C57BL/6 mice, and leukemia inhibitory factor (LIF) was used to accelerate the self-renewal, keep plasticity and inhibit differentiation to the terminal fates. The fibroblasts were exposed to five different intervention conditions:control group (DMEM+10% fetal bovine serum[FBS]) and serum deprivation (DMEM without FBS) for 24 h, 48 h and 72 h groups. qPCR was used to detect the expression of cell lineage specific genes; the in vitro angiogenesis test and Dil-Ac-LDL phagocytic function test were used to observe function of fibroblasts; and ELISA assay was used to examine the level of vascular endothelial growth factor (VEGF) secreted by fibroblasts. Results Compared with the control group, the number of capillaries-like bifurcation formed by fibroblasts was significantly increased at different serum deprivation time points (P<0.05), but no induced engulfment of Dil-Ac-LDL was noticed. Compared with the control group, the expressions of endothelial specific genes CD31 and VE-herin in fibroblasts were significantly increased at 48 h of serum deprivation (P<0.05); the expression of CD31 at 48 h of serum deprivation was 13.7 times that of the control group (P<0.05), and at 72 h of serum deprivation it was two times that of the control group (P<0.05). ELISA results showed that VEGF level at 48 h of serum deprivation was seven times that of the control group, and at 72 h of serum deprivation it was 3.7 times (P<0.05). Conclusion The serum deprivation can stimulate the transdifferentiation of cardiac fibroblasts into endothelial cell lineages.

Abstract:Objective To explore the diagnostic value of the imaging features of mural nodules in differentiating benign from malignant intraductal papillary mucinous neoplasms (IPMNs) of the pancreas. Methods In this retrospective study, we analyzed the pathology and imaging data of 112 patients with IPMNs in Changhai Hospital from Jan. 2012 to Jun. 2016. All patients underwent computed tomography (CT) and magnetic resonance imaging (MRI), and the mural nodule imaging features were observed and recorded by two resident physicians. The size, location, amount, margin and enhancement of the mural nodules were compared and analyzed in benign and malignant IPMNs. Finally, a receiver operating characteristic (ROC) curve was drawn and the area under curve (AUC) was calculated to evaluate the diagnostic value of the size, amount and margin of the mural nodules observed by imaging in differentiation of malignant and benign IPMNs, and to determine the best cut-off value, sensitivity, specificity and accuracy of the size. Results Sixty-one patients with definite mural nodules confirmed by pathology after surgery and imaging examination were included in this study; pathological analysis showed 36 patients having benign IPMNs and 25 having malignant, with main duct in 15 patients, branch duct in 13, and mixed type in 33. The malignant IPMNs mainly occurred in main duct and mixed type (P=0.01). The size of the mural nodules could be used to distinguish benign from malignant IPMNs (P<0.01), and the best cut-off value was 1.35 cm; the AUC was 74.7%, and the sensitivity, specificity and accuracy were 56.0%, 91.7% and 77.5%, respectively. Multiple mural nodules (P=0.02) and ill-defined margins (P<0.01) could be an imaging features for distinguishing benign from malignant IPMNs, and the AUC was 64.2% and 72.1%, respectively. The mural nodules of all patietns showed a progressive enhancement, which was not useful for differentiation. Conclusion The imaging features of the mural nodules are valuable for differentiation between benign from malignant IPMNs of the pancreas, and it is of clinical value for pre-operative evaluation and follow-up.

Abstract:Objective To detect the differential expression of osteopontin (OPN)-related long non-coding RNAs (lncRNAs) in sodium oxalate crystallization-induced renal tubular epithelial cell injury and to explore the role of OPN-related lncRNAs in the pathogenesis of renal injury by crystallization. Methods The renal tubular epithelial cells (HK-2 cells) were cultured and stimulated with 20 mmol/L sodium oxalate to establish the renal injury cells model. The OPN knockdown in HK-2 cell injury was carried out by transfecting OPN small interfere RNA (siRNA) into the cells, which was set up as the OPN-siRNA group; the cells in control group (NC-siRNA group) were transfected with negative random control sequences. The lncRNAs and mRNAs differential expressions were screened and identified by Arraystar Human lncRNA V4.0 Microarray in two groups, and the microarray results were analyzed by bioinformatics methods. Results Human genome lncRNA microarray results showed there were a total of 583 differentially expressed OPN-related lncRNAs in renal injury cell model of renal tubular epithelial cell induced by sodium oxalate crystallization, including 354 up-regulated lncRNAs and 229 down-regulated lncRNAs in OPN-siRNA group; there were a total of 235 mRNAs with differential expression, including 139 up-regulated mRNAs and 96 down-regulated mRNAs. Bioinformatics analysis results showed the differentially expressed lncRNAs were correlated with cell growth, enzyme activity regulation and other biological processes. Conclusion OPN-related differentially expressed lncRNAs are involved in the mechanism of the sodium oxalate crystallization-induced HK-2 cell injury.

Abstract:Objective To explore the different expression of serum proteins of the patients with breast cancer and to screen the diagnostic biomarkers. Methods Tandem mass tag (TMT)-based quantitative proteomics technology was used to detect and quantificationally analyze the serum proteins of 68 stage Ⅱ breast cancer patients and 62 healthy females; the differentially expressed proteins were screened. Bioinformatics analysis of the differentially expressed proteins were performed by UniPort Knowledgebase, Proteome Discoverer software and online software STRING (http://www.string-db.org). The protein and mRNA expressions of screened proteins, vimentin (VIME, up-regulated 3.918 folds than that of healthy females) and Raf-1 threonine-protein kinase (RAF1, down-regulated 0.251 folds) were detected by Western blotting and qPCR, respectively. Results We screened and identified 67 differentially expressed proteins, with 26 significantly up-regulated and 41 significantly down-regulated. Gene ontology (GO) and enrichment analysis showed that the differentially expressed proteins were related to tumor angiogenesis, metabolism and bioadhesive capacity. The protein and mRNA expressions of RAF1 in the patients with stage Ⅱ breast cancer were significantly lower than those in healthy females, and the protein and mRNA expressions of VIME were significantly higher, which was similar to the results of screening analysis. Conclusion TMT-based quantitative proteomics can effectively identify the differentially expressed proteins of stage Ⅱ breast cancer. VIME and RAF1 may be potential serum biomarkers of lymph node metastasis of breast cancer.

Abstract:Objective To explore the safety and efficacy of ultrasound-guided superficial cervical plexus block combined with general anesthesia in carotid endarterectomy (CEA). Methods Forty patients undergoing elective CEA were selected and divided into ultrasound-guided superficial cervical plexus block combined with general anesthesia group (combined with general anesthesia group) and simple general anesthesia group (general anesthesia group), with 20 cases in each group. We measured and analyzed the hemodynamics (invasive arterial blood pressure and heart rate), cerebral oxygen saturation, the doses of intra-operative anesthesia and vasoactive drug, post-operative visual analogue score (VAS), and one-day and three-day post-operative mini-mental state examination (MMSE) scores of the patients in the two groups at arrival to the operating room or before superficial cervical plexus block (T_0-B), after superficial cervical plexus block (T_0-L), after induction (T₁), at skin incision moment (T₂), at artery dissection moment (T₃), at carotid artery occlusion moment (T₄), at carotid artery opening moment (T₅) and two hours after operation (T₆). Results There was no significant difference in general situation between the two groups (P>0.05). In the general anesthesia group, the systolic blood pressures at T₂ and T₃ were significantly higher than that at T₁ (P<0.01), and the diastolic blood pressure, mean arterial pressure and heart rate at T₂ were significantly higher than those at T₁ (P<0.01). Except at T₄ of surgical side, the bilateral cerebral oxygen saturation at other time points in the two groups were higher than that at T_0-B, but the difference was not statistically significant (P>0.05). Compared with the general anesthesia group, the anesthesia dosage, the times of vasoactive drug and the VAS were significantly decreased in the combined with general anesthesia group (P<0.05, P<0.01), and there was no significant difference in MMSE score between the two groups (P>0.05). Conclusion The ultrasound-guided superficial cervical plexus block combined with general anesthesia can contribute to a better hemodynamic and analgesic states in CEA, and can reduce the dosage of anesthesia and vasoactive drug.

Abstract:Chemotherapy is one of the most common means of cancer treatment, and it has a lot of adverse effects and is easy to produce drug resistance. Nano-technology can be used to deliver effective dose of chemotherapeutic agents to the tumor cells and overcome tumor drug resistance, and it has no damage to normal cells. Tumor immunotherapy has the advantages of low toxicity, high effectiveness and preventing metastasis. Immunotherapy based on tumor vaccine, antibody or other immune-stimulating factor will be expected to destroy tumor and prevent tumor metastasis. Nano-technology can protect antigen and cytokine from degradation and target tumor cells, tumor interstitial or immune cells to improve the outcome. New chemical immune agents may effectively kill tumor cells as well as stimulate tumor immune antigen exposure to activate the immune system. Here, we reviewed the potential of combined application of tumor chemotherapy and immunotherapy and discussed the future of nano-technology for the targeted delivery of chemical immune agents.

Abstract:Coronary bypass grafting (CBG) is the most effective method of treating coronary artery disease and venae saphena magna is commonly used as vein graft for surgical revascularization; however, the incidence of vein graft restenosis is more than 60% in ten years after CBG, which threatens the prognosis and long-term survival of the patients. The vein graft restenosis has multiple factors and levels, including de-endothelialization, thrombosis, re-endothelialization and arterialization of vein graft in the early stage, intimal hyperplasia in the middle stage, and atherosclerosis and plaque rupture in the late stage. This review summarized the latest researches of the mechanisms of vein graft restenosis after coronary bypass grafting and discussed the prevention methods.

Abstract:Adipose-derived mesenchymal stem cells (AMDSCs) isolated from adipose tissue are stem cells with a multilineage differentiation potential, immune regulation function and highly self-renewal capacity. Compared with bone marrow mesenchymal stem cells (BMSCs), AMDSCs have some unique advantages, including abundant resource in adipose tissue, high extraction rate, and are easy to acquire tissue and so on. Hypoxic-ischemic encephalopathy (HIE) has very high mortality and disability rates, and the clinical therapeutic effect is not very satisfactory. ADMSC can decrease neurological deficits and improve neurological function following HIE through multiple mechanisms, including homing, paracrine, immune modulation, neural differentiation and endogenous neurogenesis, and the ADMSC constitute a promising therapy for HIE treatment. This review summarized the related mechanisms, clinical trials and questions about ADMSC treatment.

Abstract:Rapid hemostasis is critical for first-aid of war wound, and the application of tourniquet has always been the primary choice of combat casualties. With the change of battlefield injury form and development of hemostasis concept, the major technique for hemorrhage control has been changing from physical-aid to chemical-aid and bio-aid hemostasis, leading to the invention and application of various novel hemostatic materials and methods. The novel hemostatic techniques based on some special hemostatic agents are characterized by rapidity, convenience, high efficiency, less adverse event and good prognosis; in addition, they are particularly useful when the wounds can not be controlled by the traditional methods, such as internal hemorrhage and coagulopathy. This review summed the advance in hemostatic technology for first-aid of combat casualties, novel hemostatic agents and the related hemostatic products.

Abstract:Objective To explore the diagnostic significance of microRNA-4286 (miR-4286) for prostate cancer through detecting its expression in patients with an elevated serum prostate-specific antigen (PSA) level. Methods We selected 171 patients with an elevated serum PSA level who underwent prostate biopsy at Day Ward between Mar. 2016 and Aug. 2016 in Changhai Hospital of Second Military Medical University; we determined and compared the expression of serum miR-4286 in 50 patients with prostate cancer and 121 with benign prostate diseases by qPCR assay. The serum miR-4286 levels were analyzed at pre-operation and three-day post-operation in 10 patients with benign prostate disease and 15 patients with prostate cancer, and at pre-operation and seven-day post-operation in 8 cases with prostate cancer. The receiver operating characteristic curve was used to evaluate the diagnostic performance of miR-4286 for prostate cancer. Results The expression of miR-4286 showed a significant difference between the prostate cancer and benign prostatic disease patients (P<0.05). The expression of 3-day post-operative miR-4286 was significantly elevated compared with pre-operation in the prostate cancer patients (P<0.05); there was no significant difference in the expression of miR-4286 between pre-and 7 days post-operation in the prostate cancer patients (P>0.05) or between pre-and 3 days post-operation in the benign prostatic disease patients (P>0.05). The areas under curve of the miR-4286 and PSA were 0.653 (cut-off value 3.72, sensitivity 68.75%, specificity 60.06%) and 0.733 (cut-off value 11.38 ng/mL, sensitivity 57.16%, specificity 92.35%), respectively. Conclusion miR-4286 has a good diagnostic value and may be a new biomarker for liquid-biopsy in prostate cancer.

Abstract:Objective To investigate the correlation between the expression of △Np63 and epithelial-mesenchymal transition (EMT)-related molecular markers and its clinical significance in hypopharyngeal squamous cell carcinoma (HSCC). Methods The tumor and adjacent normal epithelium tissues of 98 patients with HSCC were collected from Department of Otorhinolaryngology Head and Neck Surgery, Changhai Hospital, Second Military Medical University from Jan. 2013 to Nov. 2016. The expression of △Np63, E-cadherin and vimentin protein in HSCC tissues, adjacent normal epithelium tissues and different regions of HSCC tissues was detected by immunohistochemistry. The correlation between the expression of △Np63, E-cadherin and vimentin proteins was examined by test of Spearman's rank-correlation coefficient. The correlation between the expression of proteins and clinicopathological features was evaluated by rank sum test. Results The expression of △Np63 and vimentin in the HSCC tissues was significantly higher than that in the adjacent normal epithelium tissues (P<0.01), while the expression of E-cadherin in the HSCC tissues was significantly lower (P<0.01). Compared with the central region of HSCC tissues, the positive expression of △Np63 and E-cadherin in the border region was significantly decreased, and the positive expression of vimentin was significantly increased (P<0.05). The expression of △Np63 was positively correlated with the expression of E-cadherin (r_s=0.409, P<0.01) and was negatively correlated with the expression of vimentin (r_s=-0.333, P<0.05) in HSCC tissues. There was no significant correlation between △Np63 and T stage, lymph node metastasis, degree of pathological differentiation or other clinicopathological features. The expression of E-cadherin and vimentin was correlated with the degree of pathological differentiation and lymph node metastasis (P<0.05). Conclusion Abnormal expression of △Np63 is associated with the occurrence and development of HSCC and the expression of EMT-related markers, indicating that △Np63 may inhibit EMT in HSCC.

Abstract:Objective To systemically evaluate the occurrence of common peri-operative complications in transcatheter aortic valve implantation (TAVI) through different approaches. Methods We searched the database, including PubMed, Embase, Ovid Medline and Database of Abstracts of Review of Effectiveness (DARE) and selected the published randomized controlled studies on the TAVI through the transapical (TAp group) or transfemoral (TF group) or transaortic (TAo group) approaches. And we compared and analyzed the 30-day post-operative mortality and the incidences of early paravalvular leakage (PVL), severe hemorrhage, major vascular complications, stroke, acute kidney injury (AKI) and permanent pacemaker (PPM) required atrioventricular block (AVB). A meta-analysis was carried out by Review Manager 5.3 software. Results A total of 22 studies, 11 530 cases were included in this study. Compared with the TF group, the incidence of early PVL in the TAp and TAo groups was significantly lower (4.6%[63/1 384] vs 9.2%[400/4 366], P<0.000 01; 6.4%[33/518] vs 9.3%[331/3 541], P=0.002), while the 30-day post-operative mortality was significantly higher (10.0%[271/2 711] vs 4.8%[326/6 756], P<0.000 01; 8.9%[46/518] vs 4.5%[160/3 541], P=0.002). The incidence of severe hemorrhage in the TAp and TAo groups was significantly higher than that in the TF group (8.4%[186/2 204] vs 3.9%[268/6 818], P<0.000 01; 6.5%[35/542] vs 1.4%[50/3 569], P=0.01). The major vascular complication had a lower incidence in the TAp group compared with the TF group (2.3%[58/2 524] vs 6.5%[417/6 367], P<0.000 01). There was no difference in the occurrence of stroke among the three groups (P>0.05). TF group showed a significant reduction of AKI incidence compared with the TAp group (8.3%[610/7 334] vs 22.8%[615/2 699], P<0.000 01) and the TAo group (3.1%[110/3 569] vs 12.5%[68/542], P<0.000 01). TF group showed a significant increase in the incidence of PPM required AVB compared with the TAo group (13.2%[472/3 569] vs 9.2%[50/542], P=0.003). Conclusion The TAp and TF approaches have a shorter path in TAVI and can significantly reduce the occurance of PVL and major vascular impairment, which is an unique advantage and has potential value.

Abstract:Objective To explore whether ghrelin can competitively inhibit glucagon-like peptide 1 (GLP-1) to stimulate the secretion of insulin via cyclic adenosinc monophosphate/protein kinase A (cAMP/PKA) pathway. Methods The pancreatic islets were isolated and purified from five 8-10 weeks old male SD rats, and identified by dithizone (DTZ) and acridine orange (AO)/propidium iodide (PI) staining. Sixty pancreatic islets were selected from each rat and then randomly divided into six groups for different disposals:S₀ group (8.3 mmol/L glucose solution), S₁ group (8.3 mmol/L glucose solution+10 nmol/L GLP-1), S₂ group (8.3 mmol/L glucose solution+10 nmol/L GLP-1+10 nmol/L ghrelin), S₃ group (8.3 mmol/L glucose solution+10 nmol/L GLP-1+10 nmol/L ghrelin+1 μmol/L growth hormone-releasing peptide 6[D-Lys³-GHRP-6], an antagonist of growth hormone secretagogue receptor 1α[GHSR-1α]), S₄ group (8.3 mmol/L glucose solution + 10 nmol/L GLP-1+10 nmol/L ghrelin+5 μmol/L forskolin, an adenylate cyclase activator), and S₅ group (8.3 mmol/L glucose solution + 10 nmol/L GLP-1+10 nmol/L ghrelin + 10 μmol/L 6-Phe-cAMP, a PKA activator); all reagents were added after last reagent treatment for 10 min. Each group had a disposal of three hours totally. ELISA assay was used to detect the concentrations of insulin and cAMP. Results The concentrations of insulin and cAMP in S₁ group were significantly higher than those in S₀ group (all P<0.05); their concentrations in S₂ group were significantly lower than those in S₁ group (both P<0.05). The concentrations of insulin and cAMP in S₃, S₄, and S₅ groups were significantly higher than those in S₂ group (all P<0.05). Conclusion Ghrelin can inhibit the effect of GLP-1 in promoting secretion of insulin, which may be mediated by cAMP/PKA pathway.

Abstract:Objective To explore the expression and clinical significance of the long non-coding RNA (lncRNA) urothelial carcinoma-associated 1 (UCA1) in the patients with intrahepatic metastasis of hepatocellular carcinoma (HCC). Methods We collected the tumor tissues, pre-and post-operative plasma samples of 20 HCC intrahepatic metastasis patients undergoing resection, the tumor tissues and plasma samples of 20 HCC patients, and the plasma samples of 20 healthy peoples from Changhai Hospital of Second Military Medical University between Jan. 2015 and Jan. 2016. qPCR was used to determine the UCA1 mRNA expression in the tumor tissues and plasma samples. We analyzed the relationship of UCA1 mRNA expression with clinicopathological characteristics, including Child-Pugh grade, the size of primary tumor, age, gender and drinking history. Results The expressions of UCA1 mRNA in the tumor tissues and the plasma of the patients with intrahepatic metastasis of HCC were significantly higher than those of HCC patients (P<0.01). In addition, UCA1 mRNA expression was significantly decreased before surgery in the patients with intrahepatic metastasis of HCC compared with after surgery (P<0.05). Expression of UCA1 mRNA in tumor tissues of HCC intrahepatic metastasis patients was correlated with the size of primary tumor (P=0.025) and Child-Pugh grade (P=0.006). Conclusion UCA1 may be a diagnostic and monitoring marker for metastasis of HCC; UCA1 expression is closely related to the development of HCC.

Abstract:Cysticercosis is often in the brain, skin, oral mucosa, tongue and other reported cases of ,but no liver cysticercosis cases .This paper reports a case of liver cysticercosis.