Objective To determine the chemical constituents in BSLYT by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS/MS),and to establish a high-performance liquid chromatographic fingerprint of the material basis of BSLYT and the content determination of its six main constituents,mononoside,loganin,oroxin A,oroxin B,baicalein and astilbin,providing a reference for the quality control.Methods The mass spectrometry data were used to establish the fingerprints.The content of the main components in the BSLYT samples was calculated by using the external method,and the discrepency between different batches of samples were analyzed by combining with chemometric methods.Results A total of 69 compounds were identified by mass spectrometry,and 13 compounds were identified after comparison with high-performance liquid chromatography(HPLC).The similarity of the baseline characterization profiles of the 15 batches of BSLYT substances was above 0.90,and a total of 27 common peaks were identified.Cluster analysis(CA)classified the substance benchmarks into 2 classes,S1,S2,S5,S8,S9,S13,and S15 were clustered into one class,and S3,S4,S6,S7,S10-S12,and S16 were clustered into one class.By combining PCA and OPLS-DA,the chemical components affecting the baseline classification of the substances were screened and attributed to wood butterfly,cornelian cherry and cocos nigra,respectively.The contents of six components were determined by MICS,which ranged from 0.31%-0.51%,0.12%-0.22%,0.09%-0.19%,0.09%-0.24%,0.07%-0.18%,and 0.08%-0.29%for mononoside,loganin,oroxin A,oroxin B,baicalein and astilbin respectively.Conclusion The fingerprint and multi-indicator content determination method established in this paper are accurate and stable,which provide a basis for the quality control of the material benchmark of Kidney and Pharynx Formula and its related preparations.

Objective: To investigate the effects CXCL1 derived from TAMs on the progression of HCC by CXCL1 in tumor microenvironment (TME) ． Methods : PMA induced human monocyte (THP-1) to obtain undifferentiated macrophages (M0) and then co-cultured with Huh 7 HCC cells．The biomarkers of macrophage phenotype and mRNA level of HCC were analysed by qRT-PCR. Colony formation assay，cell viability assay and transwell assay were performed to evaluate the biological alteration of HCC cells in TME．Epithelial-mesenchymal transition (EMT) molecular genetics were detected using western blot．The levels of CXCL1 in TME were measured by ELISA assay. Results : High CXCL1 expression in HCC patients predicted poor prognosis．Abundant macrophages were found in CXCL1 high expression HCC tissues．In macrophages and HCC co-cultured model，significantly increasing CXCL1 expression was detected in both two cells and the biomarkers of M2 macrophages，CD163 and CD206 were elevated．The growth and migration of HCC cells were promoted． Conclusion Co-culture of macrophages with HCC cells induces macrophages pro-tumor phenotype and CXCL1 secretion which promotes the progression of HCC.

Objective To investigate the application value of lymph node-targeted chemotherapy in resection of esophageal carcinoma.Methods The prospective study was conducted.The clinical data of 117 patients with middle and low esophageal carcinoma (without involving esophagogastric junction) who underwent left transthoracic esophagectomy and regional lymph node dissection in the West China Hospital of Sichuan University between January 2013 and December 2013 were collected.All patients were allocated into the 4 groups by semi-randomized control method:CPL group received intraoperatively carbon nanoparticles-paclitaxel for lymph node-targeted chemotherapy,CFL group received intraoperatively carbon nanoparticles-fluorouracil for lymph nodetargeted chemotherapy,FV group received preoperative fluorouracil intravenous chemotherapy,and control group underwent left transthoracic esophagectomy and regional lymph node dissection.CPL and CFL groups:suspensions of carbon nanoparticles and chemotherapy drugs were preoperatively prepared,and were intraoperatively injected under mucosa of lower edge of thoracic esophageal tumor using 0.1 mL syringes.FV group:fluorouracil with 100 mL of saline were mixed,and then were preoperatively injected by intravenous drip within 30 minutes.After cutting tissues of esophageal carcinoma,lymph nodes of left gastric arteria were removed,and drug level in lymph nodes was measured.At the beginning of esophagogastrostomy in the CPL,CFL and FV groups,3 mL peripheral venous blood were collected and measured for serum drug level.Control group:patients underwent left transthoractic esophagectomy and regional lymph node dissection (no blood sample and esophageal specimen).Observation indicators:(1) comparison of drug levels in lymph node and serum of patients with chemotherapy;(2) follow-up and survival:4-year cumulative survival rate in 4 groups.Follow-up using outpatient examination and telephone interview was performed to detect patients' survival up to December 2017.Measurement data with normal distribution and homogeneity of variance were represented as (x)±s,and comparisons among groups were analyzed using the ANOVA.Measurement data with skewed distribution were described as M (P25,P75),and comparisons among groups and between groups were respectively analyzed using the Kruskal-Wallis rank test and Mann-Whitney U test or Wilcoxon signed rank test.Comparisons of count data were analyzed using chi-square test or Fisher exact probability.Ordinal data were compared by the Kruskal-Wallis test.The survival curve was drawn by the Kaplan-Meier method,and Log-rank test was used for survival analysis.Results One hundred and seventeen patients were screened for eligibility,including 90 males and 27 females,and age was 37-84 years old,with an average age of 62 years old.Of 117 patients,41,41,9 and 26 were respectively allocated into the CPL,CFL,FV and control groups.Eligible patients recovered and were discharged from hospital,without bone marrow depression,severe diarrhea,anastomotic leakage and severe pneumonia.(1) Comparison of drug levels in lymph node and serum of patients with chemotherapy:drug levels in the CPL,CFL and FV groups were respectively 2.16 μg/g (1.14 μg/g,4.39 μg/g),0.44 μg/g (0.11 μg/g,1.18 μg/g),0.11 μg/g (0,0.28 μg/g) in lymph nodes and 0 (0,0),0 (0,0.31 μg/mL),0 (0,0.30 μg/mL) in serum.Drug levels of lymph node in the CPL and CFL groups were higher than those of serum,with statistically significant differences (Z=-5.579,-3.069,P＜0.05).There was no statistically significant difference in drug levels of lymph node and serum of FV group (Z =-0.365,P＞0.05).There was a statistically significant difference in drug levels of lymph node among CPL,CFL and FV groups (H=33.458,P＜0.05),and in drug levels of serum among CPL,CFL and FV groups (H=10.356,P＜0.05).Further analysis showed that fluorouracil level of lymph node in the CFL group was higher than that in the FV group,with a statistically significant difference (Z =82.500,P＜ 0.05),and there was no statistically significant difference in fluorouracil level of serum between CFL group and FV group (Z =160.500,P＞0.05).Paclitaxel level of lymph node in the CPL group was higher than fluorouracil level of lymph node in the CFL group,with a statistically significant difference (Z =351.000,P＜0.05),and paclitaxel level of serum in the CPL group was lower than fluorouracil level of serum in the CFL group,showing a statistically significant difference (Z=577.000,P＜0.05).(2) Follow-up and survival:of 117 patients,21 lost follow-up,and 96 were followed up for 6.0-58.0 months,with a median time of 20.0 months.The 4-year cumulative survival rate in the CPL,CFL,FV and control groups was respectively 46.2％,27.8％,33.3％ and 17.1％.There was no statistically significant difference in the survival of 4 groups (x2 =5.166,P＞0.05).Conclusions The lymph node-targeted chemotherapy can promote chemotherapy drugs to aggregate in the lymph nodes during resection of esophageal carcinoma.The affinity of chemotherapy drugs on carrier is involved in clinical effects,and single use of chemotherapy drug cannot improve postoperative survival rate of patients.

Objective To summarize the experience of double layered anastomosis in thoracoscopic and laparoscopic esophagectomy,and to explore its impact on the postoperative anastomotic complications.Methods Patients with thoracoscopic and laparoscopic esophagectomy from September 2014 to Auguest 2015 were retrospectively included.The cervical anastomosis were conducted by hand-sewn double layered anastomosis on the posterior wall of the gastric remnant,with the anastomotic configuration of end-to-side.The patients' general information and postoperative complications were recorded and analyzed.Results 45 patients with esophageal squamous cell carcinoma were included.The major postoperative complications were gastric dilatation(6/45,13.3％),hoarseness(5/45,11.1％),anastomotic leak/gastric necrosis(2/45,4.4％),anastomotic stricture(0/45,0).All patients were discharged from hospital with no perioperative death.Conclusion Hand-sewn double layered anastomotic technique could be safely used in thoracoscopic and laparoscopic esophagectomy,which could assure the security of the anastomosis.The anastomotic complication rates for this technique are rare enough to be recommended,as compared with other anastomotic methods reported in the literatures.

OBJECTIVETo investigate the expressions and the role of Notch signaling-associated proteins in esophageal squamous cell carcinoma (ESCC).

METHODSFifty patients with ESCC were included in this study. The expressions of Notch signaling-associated protein (4 receptors: Notch1, Notch2, Notch3, Notch4; 5 ligands: Dll1, Dll3, Dll4, Jagged1, Jagged2) in cancer foci and adjacent normal tissues (5 cm distance to cancer) were examined by immunohistochemitry. Correlations of these proteins with cancer cell proliferation(Ki-67 index) and clinicopathologic features were investigated.

RESULTSHigher levels of Notch1 and Notch2 were measured in cancer foci compared with adjacent tissues (all P<0.05). There were no differences in the expressions of Notch3, Dll1 and Dll3 (all P>0.05). Notch4, Dll4 and Jagged2 were not detected in both cancer foci and adjacent tissues. Notch1 expression was negatively correlated with lymph node metastasis and TNM staging (all P<0.01). Jagged 1 expression was positively correlated with TNM staging (P<0.01). Ki-67 index was obviously higher in cancer foci, while it was negatively correlated with Notch1 and Notch3 (all P<0.01) and positively correlated with Dll1 and Jagged 1 (all P<0.01).

CONCLUSIONNotch signaling path may act as tumor suppressive gene in the pathogenesis of esophageal squamous cell cancer, in which Notch1 protein plays an important role.

Carcinoma, Squamous Cell ; metabolism ; Esophageal Neoplasms ; metabolism ; Genes, Tumor Suppressor ; Humans ; Lymphatic Metastasis ; Neoplasm Staging ; Receptor, Notch1 ; metabolism ; Receptor, Notch2 ; metabolism ; Receptor, Notch3 ; Receptors, Notch ; metabolism ; Signal Transduction

OBJECTIVETo compare the difference of mucosal damage in the remnant esophagus with similar postoperative reflux after esophagectomy and gastric interposition between Chinese and Canadian population.

METHODSA prospective 1 to 1 paired study based on the same surgical approach was performed in Medical Centre of University of Montreal and West China Hospital of Sichuan University during the period from September 2010 to October 2013. The patients were followed up and evaluated by reflux symptom scoring, endoscopic assessment of mucosal damage, pathologic examination of biopsies and proliferation index test of esophageal epithelium.

RESULTSEighteen Han Chinese and 18 Caucasian Canadian patients with esophagectomy and gastric interposition were included in this study, with a follow-up period of 45 (28-67) months. There were no significant differences between the two groups in the incidence of postoperative reflux symptom, reflux symptom scoring, histological reflux esophagitis, erosion or stricture of remnant esophagus (all P>0.05). However, the incidence of mucosal metaplasia [44.4% (8/18) versus 11.1% (2/18), P=0.026], quantitative MUSE scoring [1.5 (1.0-2.0) versus 1.0 (0-2.0), P=0.042] and proliferation index [0.40 (0.30-0.45) versus 0.35 (0.30-0.50), P=0.038] of esophageal epithelium were significantly higher in Canadian patients than those in Chinese patients.

CONCLUSIONUnder similar reflux situation, esophageal mucosa of Canadian population is more sensitive to the gastroesophageal reflux damage compared with Chinese population, resulting in more severe reflux damage of remnant esophagus in Canadian patients.

Biopsy ; Canada ; China ; Esophagectomy ; adverse effects ; Gastroesophageal Reflux ; pathology ; Humans ; Metaplasia ; Mucous Membrane ; pathology ; Prospective Studies

Objective To compare the difference of mucosal damage in the remnant esophagus with similar postoperative reflux after esophagectomy and gastric interposition between Chinese and Canadian population. Methods A prospective 1∶1 paired study based on the same surgical approach was performed in Medical Centre of University of Montreal and West China Hospital of Sichuan University during the period from September 2010 to October 2013. The patients were followed up and evaluated by reflux symptom scoring, endoscopic assessment of mucosal damage, pathologic examination of biopsies and proliferation index test of esophageal epithelium. Results Eighteen Han Chinese and 18 Caucasian Canadian patients with esophagectomy and gastric interposition were included in this study , with a follow-up period of 45 (28-67) months. There were no significant differences between the two groups in the incidence of postoperative reflux symptom, reflux symptom scoring, histological reflux esophagitis, erosion or stricture of remnant esophagus (all P>0.05). However, the incidence of mucosal metaplasia [44.4%(8/18) versus 11.1%(2/18), P=0.026], quantitative MUSE scoring [1.5(1.0-2.0) versus 1.0 (0-2.0), P=0.042] and proliferation index [0.40 (0.30-0.45) versus 0.35 (0.30-0.50), P=0.038] of esophageal epithelium were significantly higher in Canadian patients than those in Chinese patients. Conclusion Under similar reflux situation, esophageal mucosa of Canadian population is more sensitive to the gastroesophageal reflux damage compared with Chinese population , resulting in more severe reflux damage of remnant esophagus in Canadian patients.

Objective To investigate the expressions and the role of Notch signaling-associated proteins in esophageal squamous cell carcinoma (ESCC). Methods Fifty patients with ESCC were included in this study. The expressions of Notch signaling-associated protein (4 receptors: Notch1, Notch2, Notch3, Notch4; 5 ligands: Dll1, Dll3, Dll4, Jagged1, Jagged2) in cancer foci and adjacent normal tissues (5 cm distance to cancer) were examined by immunohistochemitry. Correlations of these proteins with cancer cell proliferation ( Ki-67 index ) and clinicopathologic features were investigated . Results Higher levels of Notch1 and Notch2 were measured in cancer foci compared with adjacent tissues (all P<0.05). There were no differences in the expressions of Notch3, Dll1 and Dll3 (all P>0.05). Notch4, Dll4 and Jagged2 were not detected in both cancer foci and adjacent tissues. Notch1 expression was negatively correlated with lymph node metastasis and TNM staging (all P<0.01). Jagged 1 expression was positively correlated with TNM staging (P<0.01). Ki-67 index was obviously higher in cancer foci, while it was negatively correlated with Notch1 and Notch3 (all P<0.01) and positively correlated with Dll1 and Jagged 1 (all P<0.01). Conclusion Notch signaling path may act as tumor suppressive gene in the pathogenesis of esophageal squamous cell cancer , in which Notch1 protein plays an important role.

Objective To compare the difference of mucosal damage in the remnant esophagus with similar postoperative reflux after esophagectomy and gastric interposition between Chinese and Canadian population. Methods A prospective 1∶1 paired study based on the same surgical approach was performed in Medical Centre of University of Montreal and West China Hospital of Sichuan University during the period from September 2010 to October 2013. The patients were followed up and evaluated by reflux symptom scoring, endoscopic assessment of mucosal damage, pathologic examination of biopsies and proliferation index test of esophageal epithelium. Results Eighteen Han Chinese and 18 Caucasian Canadian patients with esophagectomy and gastric interposition were included in this study , with a follow-up period of 45 (28-67) months. There were no significant differences between the two groups in the incidence of postoperative reflux symptom, reflux symptom scoring, histological reflux esophagitis, erosion or stricture of remnant esophagus (all P>0.05). However, the incidence of mucosal metaplasia [44.4%(8/18) versus 11.1%(2/18), P=0.026], quantitative MUSE scoring [1.5(1.0-2.0) versus 1.0 (0-2.0), P=0.042] and proliferation index [0.40 (0.30-0.45) versus 0.35 (0.30-0.50), P=0.038] of esophageal epithelium were significantly higher in Canadian patients than those in Chinese patients. Conclusion Under similar reflux situation, esophageal mucosa of Canadian population is more sensitive to the gastroesophageal reflux damage compared with Chinese population , resulting in more severe reflux damage of remnant esophagus in Canadian patients.

Objective To investigate the expressions and the role of Notch signaling-associated proteins in esophageal squamous cell carcinoma (ESCC). Methods Fifty patients with ESCC were included in this study. The expressions of Notch signaling-associated protein (4 receptors: Notch1, Notch2, Notch3, Notch4; 5 ligands: Dll1, Dll3, Dll4, Jagged1, Jagged2) in cancer foci and adjacent normal tissues (5 cm distance to cancer) were examined by immunohistochemitry. Correlations of these proteins with cancer cell proliferation ( Ki-67 index ) and clinicopathologic features were investigated . Results Higher levels of Notch1 and Notch2 were measured in cancer foci compared with adjacent tissues (all P<0.05). There were no differences in the expressions of Notch3, Dll1 and Dll3 (all P>0.05). Notch4, Dll4 and Jagged2 were not detected in both cancer foci and adjacent tissues. Notch1 expression was negatively correlated with lymph node metastasis and TNM staging (all P<0.01). Jagged 1 expression was positively correlated with TNM staging (P<0.01). Ki-67 index was obviously higher in cancer foci, while it was negatively correlated with Notch1 and Notch3 (all P<0.01) and positively correlated with Dll1 and Jagged 1 (all P<0.01). Conclusion Notch signaling path may act as tumor suppressive gene in the pathogenesis of esophageal squamous cell cancer , in which Notch1 protein plays an important role.