AIM:To investigate the effects of gonadotropin-releasing hormone ( GnRH) analogue on the growth of breast cancer cell lines MCF-7 and MDA-MB-231 in vitro and to explore the related mechanisms with PI 3K/Akt or ERK/MAPK pathways .METHODS: The proliferation of human breast cancer cell lines MCF-7 and MDA-MB-231 treatment with triptorelin was detected by MTT assay and the distribution of the cell cycle was determined by flow cytometry .The phosphorylation of the ERK 1/2 and Akt was evaluated by Western blotting .RESULTS:Triptorelin inhibited the prolifera-tion of MCF-7 cells at concentration of 10-5 mol/L after treated for 192 h or at concentration of 10 -4 mol/L after treated for 168 h and 192 h.Triptorelin inhibited the proliferation of MDA-MB-231 cells at concentration of 10 -4 mol/L after treated for 192 h (P<0.05).Treatment with triptorelin for 192 h at concentration of 10 -4 mol/L had no statistical significance effect on phosphorylation of ERK1/2 and Akt(P>0.05).CONCLUSION:Inhibitory effect of GnRH analogue triptorelin on human breast cancer cells is not just the connection with the down-regulation of pituitary hormone , but also a direct in-hibitory effect.The role may not be involved in the activation of ERK /MAPK and PI3K/Akt signaling pathways .

Objective To investigate the clinical value of lasting methylene blue staining in precise hepatectomy.Methods The clinical data of 21 patients with liver cancer who received precise hepatectomy after methylene blue staining at General Hospital of PLA from February to August in 2009 were retrospectively analyzed.After the hepatic pedicle Was dissected,methylene blue WaS injected into the portal vein,and then the hepatic pedicle was ligated.Parenchymal division is initiated along the line of devascularization demarcated on Glisson capsule.Results The success rate of methylene blue staining Was 100%.Methylene blue retained in the parenchyma for(80±23)minutes.Right hepatectomy was performed on 2 patients,left hepatectomy on 1,right posterior lobectomy on 2,right anterior lobectomy on 3,left lateral lobectomy on 1,segmentectomy of segment Ⅷon 2,segmentectomy of segment Ⅶ on 3,segmentectomy of segment Ⅵ on 1,segmentectomy of segment Ⅳ on 2 and combined segmentectomy on 4.The mean volume of blood loss,incidence of postoperative complications and postoperative hospital stay were(236±6)ml,14%(3/21)and(12±3)days.Conclusions Ligation of hepatic pedicle after methylene blue injection has the advantages of high success rate and lasting staining of parenchyma of liver.Especially,this staining method contributes to improve the precision of hepatectomy by guiding the segment selection during parenchyma transection.

Objective To investigate the existence of epithelial-mesenchymal transition(EMT)and its relation to the expression of HBx protein in hepatocellular carcinoma(HCC).Methods The expression of HBx protein,E-cadherin,β-catenin,N-cadherin and fibronectin were detected by immumnohistochemistry technique in 76 cases of HCC.Results Among 76 HCC samples,the loss of expressions of E-cadherin and β-catenin were 34%(26/76)and 20%(15/76),respectively.The positive expressions of HBx protein,N-cadherin and frbronectin were observed in 68%(52/76),55%(42/76),46%(35/76)of HCC samples,respectively.The loss of expression of E-cadherin significantly correlated with the positive expression of N-cadherin and HBx protein (P＜0.01).The positive expression of N-cadherin and fibronectin significantly correlated with the loss of expression of β-catenin and the positive expression of HBx protein(P＜0.05).Conclusions EMT exists in hepatocellular carcinoma,and the expression of HBx is significantly associated with EMT.

BACKGROUND: As a kind of polypeptide, angiotensin-converting enzyme (ACE) inhibitory peptide can lower the blood pressure of human body through restraining the formation of angiotensin Ⅱ.OBJECTIVE: To investigate the influence of AGE inhibitory peptide on endothelial cell proliferation and endothelin expression in cultured human umbilical vein cells based on cellular and molecular levels in order to provide the experimental evidences for ACE inhibitory peptide to be the potential blood pressure-lowering health food.DESIGN: Repeated measures design.SETTING: School of Bioscience and Bioengineering, South China University of Technology; School of Applied Chemistry and Biological Technology,Shenzhen Polytechnic.MATERIALS: The experiment was carried out in the Institute of Applied Chemistry and Biological Technology, Shenzhen Polytechnic from September 2004 to March 2005. The AGE inhibitory peptide was provided by the Institute of Applied Chemistry and Biological Technology, Shenzhen Polytechnic. Under certain circumstance, 15 μ mol/L of the inhibitor was needed to decrease half of the AGE activity. The human umbilical vein endothelial cells of the 4th generation were cultured randomly in 7 groups with different concentrations: medium group, 150, 300 and 600 mg/L ACE inhibitory peptide groups, captopril group, norepinephrine(NE) group, and ACE inhibitory peptide+NE group.METHODS: ①The endothelial cells were cultured as recommended. The medium was M199+FBS(0.15, v/v)+penicillin(10 000 U/mL)+streptomycin (100 mg/L). After cellular fusion, the cells were carried on the passage with the ratio of 1:2. The 4th generation cells were used for experiment. ②M199(0.15, v/v) was contained in each group. ACE inhibitory peptides were added to make the final concentration 150, 300 and 600 mg/L in the 150, 300 and 600 mg/L ACE inhibitory peptide groups respectively. Captopril was added to make the final concentration 10-5 mol/L in the captopril group. NE was added to make the final concentration 100 μg/L in the NE group. ACE inhibitory peptide and NE were added to make the final concentration 300 mg/L and 100 μg/L in the ACE inhibitory peptide+NE group respectively. ③The state of cell growth was determined with cytometry. The contents of endothelial cells in the medium with different culture times were determined with radioimmunoassay. The expression of endothe lin mRNA was determined with reverse transcriptase-polymerase chain reaction. The expression of cellular endothelin protein was determined with immunohistochemical method.MAIN OUTCOME MEASURES: ①The influence of ACE inhibitory peptide on endothelial cell proliferation. ②The influence of ACE inhibitory peptide on the endothelin mRNA and endothelin protein.RESULTS: ①The influence of ACE inhibitory peptide on endothelial cell proliferation and endothelin secretion: Compared with the medium group,in the captopril and 150, 300, 600 mg/L ACE inhibitory peptide groups,the growth of endothelial cells was restrained and the endothelin content in the medium was lowered(P ＜ 0.01 or 0.05). NE could promote the growth of endothelial cells and the secretion of endothelin, but the cell density and endothelin content after treatment with ACE inhibitory peptide were similar to those in the medium group (P ＞ 0.05). ②The influence of ACE inhibitory peptide on the expressions of endothelin mRNA and protein in endothelial cells: Compared with the medium group, the expressions of endothelin mRNA and protein might be lowered in the captopril and 150,300, 600 mg/L ACE inhibitory peptide groups(P ＜ 0.01 or 0.05). The expressions of endothelin mRNA and protein could be up-regulated by NE.The gene expression after treatment with ACE inhibitory peptide was similar to that in the medium group(P ＞ 0.05).CONCLUSION: The ACE inhibitory peptides of different dosages can all restrain the growth of endothelial cells, lower the endothelin content, decrease the expression of endothelin gene and resist NE improved growth and secretion of endothelial cells in umbilical vein cell effectively.

Objective:To investigate the prognostic value of preoperative inflammatory indicators for hepatocellular carcinoma (HCC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 73 patients with primary HCC who underwent radical partial hepatectomy in the Beijing Tsinghua Changgung Hospital of Tsinghua University from December 2014 to July 2019 were collected. There were 57 males and 16 females, aged from 33 to 81 years, with a median age of 58 years. Results of blood examination indicators at the first time in hospital were determined for patients. Observation indicators: (1) the best cut-off values of?? preoperative inflammatory indicators calculated by the maximally selected rank statistics; (2) follow-up; (3) influencing factors for prognosis of HCC patients; (4) comparison of clinicopathological parameters of HCC patients; (5) comparison of predictive value for overall survival. Follow-up was conducted using outpatient examination and telephone interview to determine postoperative survival of patients up to September 2019. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). The best cut-off values ??for continuous variables were obtained using the maximally selected rank statistics based on survival at endpoint of follow-up. Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Kaplan-Meier method was used to calculate survival rates, and Log-rank test was used for survival analysis. Univariate analysis was performed using the Log-rank test. Multivariate analysis was performed using the COX proportional hazard model. The time-dependent receiver operating characteristic curve (ROC) was used to compare the predictive value of independent prognostic factors. Results:(1) The best cut-off values of?? preoperative inflammatory indicators calculated by the maximally selected rank statistics: the best cut-off values of neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), and prognostic nutrition index (PNI) were 3.46, 131.05, and 45.65. (2) Follow-up: 73 patients were followed up for 31 months (range, 2-57 months). Twenty patients died during the follow-up. (3) Influencing factors for prognosis of HCC patients: results of univariate analysis showed that NLR, PNI, tumor diameter, and tumor differentiation degree were related factors affecting prognosis of patients ( χ2=10.213, 4.434, 5.174, 4.306, P<0.05). Results of multivariate analysis showed that NLR and tumor differentiation degree were independent factors affecting prognosis of patients ( hazzard ratio=4.429, 13.278, 95% confidence interval as 1.662-11.779, 1.056-10.169, P<0.05). (4) Comparison of clinicopathological parameters of HCC patients: of 73 patients, 64 cases had NLR<3.46 and 9 cases had NLR≥3.46. Cases with tumor length >5 cm or ≤5 cm, neutrophils, lymphocytes were 23, 41, (2.9±1.2)×10 9/L, (1.7±0.6)×10 9/L for 64 patients with NLR<3.46, versus 8, 1, (5.8±2.9)×10 9/L, (1.0±0.3)×10 9/L for 9 patients with NLR≥3.46; there were significant differences in above indicators between the two groups ( χ2=7.017, t=2.982, -3.168, P<0.05). (5) Comparison of predictive value for overall survival: time-dependent ROC curves of NLR and tumor differentiation degree for 1-, 2-, 3-, 4-year survival rates had the area under curve of 0.735,0.611, 0.596, 0.574 and 0.554, 0.583, 0.572, 0.556, respectively. NLR had better predictive value for overall survival of patients than tumor differentiation degree. Conclusion:Preoperative NLR is an independent factor affecting prognosis patients, and its predictive efficacy is better than tumor differentiation degree.

For unresectable advanced hepatocellular carcinoma (HCC),besides sorafenib,alternative drugs and treatment modalities are required.Clinical studies of hepatic arterial infusion chemotherapy (HAIC),transcatheter arterial chemoembolization (TACE),and system chemotherapy have shown favorable efficacy and tolerance in advanced HCC patients.In addition,the potential efficacy of sorafenib combined with focal treatment is also an interesting issue.As more therapies become available,decision-making for treating advanced HCC becomes increasingly complex.In our opinion,diverse treatment modalities should be utilized for the best interest of patients.Based on predictive biomarkers,we should develop a precise patient stratification system to select suitable candidates for each treatment modality in future studies,as is useful for improving prognosis of patients with advanced HCC.

Objective To investigate the value and efficacy of continuous renal replacement therapy(CRRT) for treatment of heat stroke patients complicated by multiple organ dysfunction syndrome(MODS). Methods The clinical data of 19 heat stroke patients complicated by MODS admitted into the hospital in a period from July 15,2010 to August 30,2010 and treated by CRRT were analyzed retrospectively. Continuous venovenous hemofiltation(CVVH) mode was used in all patients and the initial temperature of replacement fluid range was 28℃to 32℃persisting in 2.0 to 2.5 hours and afterward it maintained at 36℃. Prognosis and adverse effect were observed,the patients' body temperature,heart rate(HR),mean arterial pressure(MAP),acute physiology and chronic health evaluationⅡ(APACHEⅡ)scores,oxygenation index(PaO2/FiO2),the levels of serum urea nitrogen(BUN),serum creatinine(SCr), myoglobin(Mb),creatine kinase(CK),alanine aminotransferase(ALT),aspartate aminotransferase(AST)and arterial lactate(Lac)were monitored before and after CRRT treatment. Results Fifteen patients were cured or improved,and 4 died. Compared with those before CRRT treatment,body temperature(℃),HR(bmp),MAP(mm Hg,1 mm Hg=0.133 kPa),APACHEⅡevaluation(score),PaO2/FiO2(mm Hg)were significantly improved(body temperature:36.8±0.2 vs. 41.6±0.3,HR:93.6±10.3 vs. 132.5±11.4,MAP:69.8±9.9 vs. 45.2±7.7,APACHEⅡ:12.3±3.9 vs. 29.6±4.6,PaO2/FiO2:213.6±95.4 vs. 126.5±87.4,all P<0.05);the levels of BUN(mmol/L),SCr(μmol/L), Mb(μg/L),CK(U/L),ALT(U/L),AST(U/L),Lac(mmol/L)were significantly reduced after the treatment(BUN:23.9±5.3 vs. 42.6±5.4,SCr:123±47 vs. 356±51,Mb:201±45 vs. 468±39,CK:217±32 vs. 843±41,ALT:79±36 vs. 894±88,AST:57±28 vs. 867±92,Lac:3.5±2.4 vs. 16.6±3.9,all P<0.05). In the process of the treatment,hemodynamics was stable,and no obvious side effects occurred. Conclusion CRRT treatment can exactly and safely reduce the core body temperature of patients with heat stroke,and it can also effectively eliminate metabolites of BUN,Cr,Mb,etc,ameliorate the inflammatory reaction and supporting the functions of liver,kidneys and other vital organs,thus the treatment is also safe and effective for such patients complicated by MODS.

Objective To evaluate three-dimentional (3D) reconstruction technique and methylene blue staining in precise anatomic hepatectomy.Methods The clinical data of 12 patients with hepatocellular carcinoma who were admitted to the Chinese PLA General Hospital from February 2009 to August 2011 were retrospectively analyzed.The 3D reconstruction of the liver tumor and intrahepatic vessels were done based on the computed tomography data and magnetic resonance imaging data.The portal vein supplying the tumor and its anatomic relationship with adjacent vessels were evaluated.Precise anatomic hepatectomy was performed guided by sustained methylene blue staining.Results The accurate rate of 3D model of the portal triad was 12/12.The shape of target segments observed after methylene blue staining was consistent with the results of 3D evaluation.Two patients received hemihepatectomy,3 received lobectomy,5 received monosegmentectomy or subsegmentectomy,2 received multisegmentectomy.The mean tumor diameter,operation time,blood loss,postoperative hospital stay and complication rate were 5.6cm (2.5-16.0 cm),(150±24)minutes,(236±25)ml,(10±3)days and 2/12,respectively.After a median follow-up of 14 months,tumor recurrence was found in 2 patients,and 1 of them died of tumor progression.Conclusions The 3D reconstruction may contribute to precise evaluation of the anatomic relationship between the tumor and its adjacent vessels.The 3D technique combined with sustained methylene blue staining may significantly improve the accuracy of anatomic hepatectomy.

Objective To examine the role of c-Src activation in hepatitis B virus X (HBx) protein induced epithelial-mesenchymal transition (EMT) in liver cancer.Methods SMMC-7721 liver cancer cells were transfected with HBx gene to induce EMT and the activated c-Src expression was evaluated by Western blot.Both the morphological changes and the epithelial and mesenchymal markers expression (real-time PCR,western blot and immunocytochemistry) of HBx-transfected SMMC-7721 cell treated by c-Src kinase inhibitor PP2 and negative control PP3 were observed and compared,respectively.Results The activated c-Src expression in HBx gene transfected SMMC-7721 cells was significantly increased compared to that in mock transfected cells,c-Src kinase inhibitor PP2 could enable the HBx-transfected SMMC-7721 cells to transmit from spindle-like shape to original epithelial morphology.Western blot and immunocytochemistry confirmed that the expression of epithelial markers and mesenchymal markers almost returned to the levels of parental cells,indicating the mesenchymal-epithelial transition.Conclusions c-Src activation plays a key role in the process of EMT induced by HBx protein in SMMC-7721 cells.

Objective To investigate the clinical value of computer-assisted operation planning system for precise hepatectomy.Methods The clinical data of 45 patients who had undergone precise hepatectomy at General Hospital of PLA from November 2006 to November 2009 were retrospectively analyzed.The 3-dimensional imaging of liver was constructed by computer-assisted operation planning system,and the anatomic relationship between hepatic vasculature and tumor Was analyzed.Volume of hepatic segments,total liver volume,tumor volume,predicted resection volume and the remnant liver volume were calculated before operation.Virtual liver resection was performed in order to select the best operative procedure.The predicted resection volume Was compared with the hepatic segment resected.All data were analyzed by t test,Pearson rank correlation analysis and chi-square test.Results The predicted resection volume calculated based on the 2-dimensional imaging was similar to that calculated based on the 3-dimensional imaging(t=2.125,P>0.05).The three-grade branches of the hepatic artery,portal vein and hepatic vein were clearly shown in the computer-assisted operation planning system.The anatomic relationship between tumor and adjacent vessels was quantitatively analyzed.There was a positive correlation between the predicted resection volume and the resected liver volume(r=0.999,P<0.05),and the error rate Was 5.1%.All patients received anatomical hepatectomy,and the incidence of postoperative complications was 20%(9/45).No liver failure or perioperative mortality was observed.Conclusion Computer-assisted operation planning system may facilitate and promote precise hepatectomy.