1.Parthenolide enhances the apoptosis induced by PKC inhibitor in human gastrointestinal stromal tumors cell lines
Xiangdan LI ; Lan LIU ; Xuesen FANG ; Toufeng JIN
Basic & Clinical Medicine 2017;37(2):202-205
Objective To investigate the effect of parthenolide ( PTL) and PKC inhibitor on human gastrointestinal stromal tumor (GIST) cell proliferation and apoptosis and the mechanism involved .Methods Human GIST cell lines were cultured in vitro, and the cell proliferation rate of GIST , was determinate by MTT;flow cytometry was used to test the early apoptosis rate of GIST;Western blot assay was applied to detect the expression of endoplasmic reticulum stress-related proteins , GRP78 and GADD153.There were four groups: control group , PTL group, PKC inhibitor group , combine PTL and PKC inhibitor group .Results PTL and PKC inhibitor combination therapy for GIST was sig-nificantly more effective than a single-drug therapy (P<0.05);as for the early apoptosis rate , the combination ther-apy for GIST cells was significantly higher than that medication alone group (P<0.05).the expression of endoplas-mic reticulum stress-associated protein GRP 78 and GADD153 was obviously higher in PTL and PKC inhibitor combi-nation group than that in medication alone group (P<0.05).Conclusions PTL and PKC inhibitor combination therapy for GIST cells can induce apoptosis , which is possibly mediated via endoplasmic reticulum stress pathway .
2.Experimental in vivo and in vitro study of inhibitory effect of extract from taraxacum mogonon on human poorly differentiated gastric cancer
Yucheng SUN ; Xuesen FANG ; Chunxiao LIE
Cancer Research and Clinic 2018;30(2):79-84
Objective To investigate inhibitory effect and mechanism of extract from taraxacum mogonon (ETM) on human poorly differentiated gastric cancer MKN45 cells and their model nude mice tumors. Methods Human poorly differentiated gastric cancer MKN45 cells cultured in vitro were divided into the control group and the experimental group. The experimental group was treated with different concentrations of ETM, while the control group was treated with phosphate buffered saline (PBS). The tumor cell growth was examined by methyl thiazolyl tetrazolium(MTT).Cell apoptosis was detected by flow cytometry (FCM). The expression of Survivin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR), and the protein expression of Survivin was detected by Western blot. The nude mouse model with poorly differentiated gastric cancer was established with MKN45 cells. Successful models of nude mice were divided into the experimental group and the control group according to random number table. ETM 0.4 ml (100 mg/ml) was injected into abdominal cavity of the nude mice in the experimental group, while only equal dose of 0.9 % NaCl solution was injected in the control group, every 3 days for once, 42 days in total. The weight and volume changes of transplantable tumors were observed. The protein expression of Survivin in transplantable tumors was examined by immunohistochemistry(IHC) method. Results ETM could inhibit the cell proliferation and promote apoptosis of MKN45 cells in a dose-dependent and time-dependent way when its concentration reached 0.2 μmol/L (P< 0.05). The expression of Survivin mRNA and Survivin protein was down regulated in a dose-dependent way in the experimental group. Survivin mRNA expression in the experimental group with 0.1,0.2,0.4,0.8, 1.6 μmol/L of ETM and in the control group was 0.18±0.04, 0.14± 0.03, 0.11±0.01, 0.08±0.04, 0.06±0.02 and 0.19±0.03 respectively (F = 132.35, P< 0.05); Survivin protein expression in the experimental group in 0.1, 0.2, 0.4, 0.8, 1.6 μmol/L of ETM, and in the control group was 0.86±0.03, 0.60±0.05, 0.43±0.01, 0.22±0.01, 0.14±0.03 and 0.92±0.06 respectively (F= 76.57, P< 0.05). The difference of experimental group was statistically significant when concentration of ETM reached 0.2 μmol/L or above compared with the control group (P< 0.05). Nude mouse experiment showed that the tumor inhibition rate was (60.3±3.2) % in the experimental group. The volume and weight of transplantable tumor in the experimental group was(326± 27)mm3and(0.31±0.13)g,which was obviously lower than that in the control group[(843±14) mm3and (0.78±0.25) g]. The difference was statistically significant (t= 3.94,P =0.043;t= 3.27,P = 0.037). IHC experiment results showed that positive cell count of Survivin protein in the experimental group was obviously less than that in the control group(28±11 vs.152±20;t =4.32,P =0.029). Conclusions ETM has an obvious inhibitory effect on human poorly differentiated gastric cancer MKN45 cell and its nude mouse tomor model. The mechanism may be related with down-regulation of Survivin expression caused by ETM.
3.The pulmonary arterial hypertension of patients on maintained hemodialysis is positively associated with the decreased percent of CD8 T cell in the peripheral blood independently
Rongyi CHEN ; Fangfang XIANG ; Jiachang HU ; Xuesen CAO ; Yi FANG ; Bo SHEN ; Zhonghua LIU ; Wenlyu LYU ; Xiao TAN ; Zhihui LU ; Xiaoqiang DING ; Jianzhou ZOU
Chinese Journal of Nephrology 2017;33(5):342-348
Objective To explore the risk factors of pulmonary artery hypertension (PAH) and the its relationship with T cell subsets to provide a foundation for the prevention and treatment of PAH.Methods 154 maintained hemodialysis (MHD) patients in our dialysis center were recruited according to the criterion and divided into two groups subsequently:PAH group (pulmonary artery systolic pressure,PASP > 35 mmHg) and non-PAH group (PASP≤35 mmHg).The related clinical,biochemical and ultrasonic cardiogram data were collected and peripheral blood was acquired to detect the expressions of the surface antigen CD3,CD4,CD8 and CD69 with flow cytometry.Logistic regression analysis was used to find out the relationship between PAH and T cell subsets.Results There was no significant difference between 56 cases of PAH and 98 cases of non-PAH as regards gender,age,mean systolic and diastolic pressure,dialysis durations,morbidities of hypertension and diabetes,smoking rate,and left ventricular diameter.Compared with the non-PAH group,the PAH group demonstrated a lower percent of CD8 T cells and CD8 CD69 T cells,but a much higher left atrial diameter (LAD),Interventricular septum thickness,left ventricular posterior wall thickness,and NT-proBNP.The percentage of T cells,CD4 T cells and CD4 CD69 T cells showed no difference between the two groups.Multivariate analysis confirmed that PAH was negatively independently associated with the percentage of CD8 T cells and CD8CD69 T cells.Conclusions The decreased percentage of CD8 T cells and CD8CD69 T cells in the peripheral blood is a risk factor of PAH in maintained hemodialysis patients,and CD8 T cells may play an important role in the genesis of PAH.
4.Effect of dynamic lung compliance-guided PEEP titration on lung injury in elderly patients undergo-ing robot-assisted radical prostatectomy
Xin YUAN ; Shouyuan TIAN ; Xin WANG ; Lixia NIE ; Xuesen SU ; Rui ZHANG ; Chao LI ; Shu-Fang LIU
Chinese Journal of Anesthesiology 2019;39(3):264-267
Objective To evaluate the effect of dynamic lung compliance ( Cydn)-guided positive end-expiratory pressure (PEEP) titration on lung injury in the patients undergoing robot-assisted radical prostatectomy. Methods Forty American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients, aged 65-80 yr, with body mass index of 19-28 kg∕m2 , scheduled for elective robot-assisted radical prosta-tectomy under general anesthesia, were divided into 2 groups ( n=20 each) using a random number table method: control group (group C) and PEEP group (group P). Mechanical ventilation was performed ac-cording to preset parameters after tracheal intubation in group C, and PEEP was set by a double titration method after tracheal intubation and after pneumoperitoneum in group P. At 4 min after intubation (T1), 4 min, 1 h and 2 h after establishing pneumoperitoneum-Trendelenburg position ( T2-4 ) , and 1 and 30 min after extubation ( T5,6 ) in group C or at 4 min after completing the first PEEP titration ( T1 ) , 4 min, 1 h and 2 h after completing the second PEEP titration (T2-4) and T5,6 in group P, blood samples were collect-ed from the radial artery for determination of club cell protein 16, surfactant protein-D, tumor necrosis fac-tor-alpha and interleukin-6 concentrations in serum ( by enzyme-linked immunosorbent assay) . Results Compared with group C, the serum concentrations of club cell protein 16 at T2-6 and surfactant protein-D, tumor necrosis factor-alpha and interleukin-6 at T3-6 were significantly decreased in group P (P<0. 05). Conclusion Cydn-guided PEEP titration can reduce the lung injury in patients undergoing robot-assisted radical prostatectomy.
5.Effect of dynamic lung compliance-guided PEEP titration on extravascular lung water in elderly pa-tients undergoing robot-assisted radical prostatectomy
Xuesen SU ; Shouyuan TIAN ; Xin WANG ; Lixia NIE ; Xin YUAN ; Rui ZHANG ; Chao LI ; Shu-Fang LIU
Chinese Journal of Anesthesiology 2019;39(4):415-417
Objective To evaluate the effect of dynamic lung compliance ( Cdyn) -guided positive end-expiratory pressure (PEEP) titration on extravascular lung water in elderly patients undergoing robot-assisted radical prostatectomy. Methods Forty American Society of Anesthesiologists physical statusⅡ orⅢ patients, aged 65-80 yr, with body mass index of 19-28 kg∕m2 , scheduled for elective robot-assisted radical prostatectomy, were divided into 2 groups ( n=20 each) using a random number table method:control group (group C) and PEEP group (group P). In group P, immediately after endotracheal intuba-tion, immediately after establishing pneumoperitoneum-Trendelenburg position and after restoring the supine position, PEEP was set starting from the lowest PEEP allowed by the machine, increasing by 2 cmH2 O ev-ery 4 min until the maximum Cdyn was obtained. PEEP was not set in group C. Immediately after establis-hing the invasive blood pressure monitoring ( T1 ) , at 10 min after the first successful PEEP titration ( T2 ) , 10 min, 1 h and 2 h after the second successful PEEP titration ( T3-5 ) , 10 min after the third successful PEEP titration (T6), and 30 min after tracheal extubation (T7) in group P, or at T1, 10 min after intu-bation ( T2 ) , 10 min, 1 h and 2 h after establishing pneumoperitoneum-Trendelenburg position ( T3-5 ) , 10 min after restoring the supine position ( T6 ) and T7 in group C, blood samples were collected from the radial artery for blood gas analysis, and the oxygenation index was calculated. The B-line score was recor-ded at T1 and T7 . Results Compared with group C, the B-line score was significantly decreased at T7 , and the oxygenation index was increased at T5-7 in group P (P<0. 05). Conclusion Cdyn-guided PEEP titration can decrease the formation of extravascular lung water in elderly patients undergoing robot-assisted radical prostatectomy.
6.Therapeutic strategies for a functional cure of chronic hepatitis B virus infection.
Jinhong CHANG ; Fang GUO ; Xuesen ZHAO ; Ju-Tao GUO
Acta Pharmaceutica Sinica B 2014;4(4):248-257
Treatment of chronic hepatitis B virus (HBV) infection with the viral DNA polymerase inhibitors or pegylated alpha-interferon has led to a significant retardation in HBV-related disease progression and reduction in mortality related to chronic hepatitis B associated liver decompensation and hepatocellular carcinoma. However, chronic HBV infection remains not cured. The reasons for the failure to eradicate HBV infection by long-term antiviral therapy are not completely understood. However, clinical studies suggest that the intrinsic stability of the nuclear form of viral genome, the covalently closed circular (ccc) DNA, sustained low level viral replication under antiviral therapy and homeostatic proliferation of hepatocytes are the critical virological and pathophysiological factors that affect the persistence and therapeutic outcomes of HBV infection. More importantly, despite potent suppression of HBV replication in livers of the treated patients, the dysfunction of HBV-specific antiviral immunity persists. The inability of the immune system to recognize cells harboring HBV infection and to cure or eliminate cells actively producing virus is the biggest challenge to finding a cure. Unraveling the complex virus-host interactions that lead to persistent infection should facilitate the rational design of antivirals and immunotherapeutics to cure chronic HBV infection.