1.Trichostatin A up-regulates coxsackievirus-adenovirus receptor (CAR) expression through inhibiting MAPK/ERK pathway to enhance the antitumor activity of H101 virus in thymic carcinoma
Zhanfeng HE ; Wei WANG ; Tianliang ZHENG ; Donglei LIU ; Yang YANG ; Dengyan ZHU ; Kai WU ; Liping WANG ; Song ZHAO
Chinese Journal of Microbiology and Immunology 2020;40(8):628-634
		                        		
		                        			
		                        			Objective:To investigate the expression of coxsackievirus-adenovirus receptor (CAR) in thymic carcinoma and the relationship between CAR and the antitumor activity of oncolytic adenovirus H101.Methods:The expression of CAR in thymic carcinoma tissues and cells were detected by RT-qPCR and Western blot. H101 expression and virus titers in Bcap-37, MP59 and T1889 cells after infection were detected by RT-qPCR and 50% tissue culture infectious dose (TCID 50). The proliferation activity and apoptosis rates of T1889 cells infected with H101 at different multiplicity of infection (MOI) were detected by CCK-8 and flow cytometry. CAR expression in T1889 cells treated with different concentrations of trichostatin A (TSA), a histone deacetylase inhibitor, was detected. H101 expression and virus titers in the TSA-treated and H101-infected cells were detected. Cell activity was detected by CCK-8. The phosphorylation levels of MARK and ERK1/2 and the expression of CAR at protein level in TSA-treated or TSA+ TBHQ (ERK activator) treated cells were detected. Results:CAR expression at both mRNA and protein levels were significantly lower in thymic carcinoma tissues than in adjacent normal tissues ( P<0.01), and lower in MP59 and T1889 cells than in thymic epithelial cells (TEC) and Bcap-37 cells ( P<0.01). H101 expression in MP59 and T1889 cells and the titers of H101 in culture supernatants were significantly lower than those in Bcap-37 cells ( P<0.01). Compared with Bcap-37 cell, the activity of MP59 and T1889 cells was significantly increased and the apoptosis rates were significantly decreased 48 h after H101 infection ( P<0.01). The expression of CAR at both mRNA and protein levels in T1889 cells treated with different concentrations of TSA increased in a dose-dependent manner. When T1889 cells were treated with 0.25 μmol/L of TSA, the expression of H101 at mRNA level and H101 titers were significantly increased ( P<0.05); the phosphorylation levels of MAPK and ERK1/2 proteins were continuously decreased; the expression of CAR was continuously increased. Compared with the TSA treatment group, the expression of CAR at protein level in the TSA+ TBHQ treatment group decreased significantly ( P<0.01), and the p-ERK1/2/ERK1/2 ratio increased significantly ( P<0.01). Conclusions:TSA could up-regulate CAR expression in thymic carcinoma by inhibiting the MARK/ERK1/2 pathway, thereby enhancing the antitumor activity of H101.
		                        		
		                        		
		                        		
		                        	
2.Comparison of short-term and long-term efficacy between robot-assisted and thoracoscopy-laparoscopy-assisted radical esophageal cancer surgery
Zhanfeng HE ; Tianliang ZHENG ; Donglei LIU ; Yang YANG ; Dengyan ZHU ; Kai WU ; Liping WANG ; Song ZHAO
Chinese Journal of Gastrointestinal Surgery 2020;23(4):390-395
		                        		
		                        			
		                        			Objective:To explore the safety and short-term and long-term efficacy of robot-assisted radical esophageal cancer surgery.Methods:A prospective randomized controlled trial was conducted. Patients who were preoperatively diagnosed as stage 0-IIIB esophageal squamous cell carcinoma and suitable for minimally invasive surgery in our hospital from January 1, 2014 to June 30, 2018 were prospectively enrolled. Those of age ≥75 years having received preoperative neoadjuvant therapy, contradicted to anesthesia or operation due to severe complications, with history of thoracotomy or laparotomy, with concurrent malignant tumors, without complete informations or refusing to participate in this study were excluded. Participants were randomly divided into the thoracoscopy-laparoscopy group and the robot group using a random number table in ratio of 1:1. Preoperative clinicopathological data, surgical data and postoperative outcomes were recorded. The patients were followed up mainly by telephone. Follow-up endpoint was recurrence of esophageal cancer and death. Kaplan-Meier method was used to estimate survival rate. The survival difference between the two groups was analyzed using the log-rank test.Results:According to above criteria, a total of 192 esophageal cancer patients were enrolled finally, including 144 males and 48 females with mean age of (61.9±8.6) years. The robot group had 94 cases, including 72 males and 22 females with mean age of (61.3±8.2) years, and the thoracoscopy-laparoscopy group had 98 cases, including 72 males and 26 females with mean age of (62.4±9.1) years. There were no significant differences in baseline data between the two groups (all P>0.05). Operation was abandoned in one case in each group due to extensive pleural cavity metastasis and one case in each group was converted to thoracotomy. The success rate of operation was 97.9% (92/94) in the robot group and 98.0% (96/98) in the thoracoscopy-laparoscopy group (χ 2=0.002, P=0.996). The number of lymph nodes dissected in the robot group was significantly higher than that in the thoracoscopy-laparoscopy group (29.2±12.5 vs. 22.8±13.3, t=3.433, P=0.001), while there were no significant differences in operative time, intraoperative blood loss, R0 resection rate, postoperative 30-day mortality, postoperative hospital stay, ICU stay, time to withdrawal of chest drainage tube, ICU readmission, and postoperative morbidity of complications between the two groups (all P>0.05). The median follow-up time was 21 (3 to 57) months. During the follow-up, 3 cases and 4 cases were lost, and 2 cases and 3 cases died of other diseases in the robot group and in the thoracoscopy-laparoscopy group respectively. Recurrence occurred in 39 cases during follow-up, including 14 recurrences in the robotic group with 1- and 3-year recurrence-free survival rates of 92.4% and 87.6% respectively and the median recurrence time of 15 (9 to 42) months. There were 25 recurrences in the thoracoscopy-laparoscopy group with 1- and 3-year recurrence-free survival rates of 81.7% and 67.9% respectively and the median recurrence time of 9 (3 to 42) months. There was significant difference in recurrence-free survival between the two groups (χ 2=4.193, P=0.041). Conclusions:The robotic surgical system has good oncology effect and surgical safety in the radical operation of esophageal cancer, which deserves further research and promotion.
		                        		
		                        		
		                        		
		                        	
3.Comparison of short-term and long-term efficacy between robot-assisted and thoracoscopy-laparoscopy-assisted radical esophageal cancer surgery
Zhanfeng HE ; Tianliang ZHENG ; Donglei LIU ; Yang YANG ; Dengyan ZHU ; Kai WU ; Liping WANG ; Song ZHAO
Chinese Journal of Gastrointestinal Surgery 2020;23(4):390-395
		                        		
		                        			
		                        			Objective:To explore the safety and short-term and long-term efficacy of robot-assisted radical esophageal cancer surgery.Methods:A prospective randomized controlled trial was conducted. Patients who were preoperatively diagnosed as stage 0-IIIB esophageal squamous cell carcinoma and suitable for minimally invasive surgery in our hospital from January 1, 2014 to June 30, 2018 were prospectively enrolled. Those of age ≥75 years having received preoperative neoadjuvant therapy, contradicted to anesthesia or operation due to severe complications, with history of thoracotomy or laparotomy, with concurrent malignant tumors, without complete informations or refusing to participate in this study were excluded. Participants were randomly divided into the thoracoscopy-laparoscopy group and the robot group using a random number table in ratio of 1:1. Preoperative clinicopathological data, surgical data and postoperative outcomes were recorded. The patients were followed up mainly by telephone. Follow-up endpoint was recurrence of esophageal cancer and death. Kaplan-Meier method was used to estimate survival rate. The survival difference between the two groups was analyzed using the log-rank test.Results:According to above criteria, a total of 192 esophageal cancer patients were enrolled finally, including 144 males and 48 females with mean age of (61.9±8.6) years. The robot group had 94 cases, including 72 males and 22 females with mean age of (61.3±8.2) years, and the thoracoscopy-laparoscopy group had 98 cases, including 72 males and 26 females with mean age of (62.4±9.1) years. There were no significant differences in baseline data between the two groups (all P>0.05). Operation was abandoned in one case in each group due to extensive pleural cavity metastasis and one case in each group was converted to thoracotomy. The success rate of operation was 97.9% (92/94) in the robot group and 98.0% (96/98) in the thoracoscopy-laparoscopy group (χ 2=0.002, P=0.996). The number of lymph nodes dissected in the robot group was significantly higher than that in the thoracoscopy-laparoscopy group (29.2±12.5 vs. 22.8±13.3, t=3.433, P=0.001), while there were no significant differences in operative time, intraoperative blood loss, R0 resection rate, postoperative 30-day mortality, postoperative hospital stay, ICU stay, time to withdrawal of chest drainage tube, ICU readmission, and postoperative morbidity of complications between the two groups (all P>0.05). The median follow-up time was 21 (3 to 57) months. During the follow-up, 3 cases and 4 cases were lost, and 2 cases and 3 cases died of other diseases in the robot group and in the thoracoscopy-laparoscopy group respectively. Recurrence occurred in 39 cases during follow-up, including 14 recurrences in the robotic group with 1- and 3-year recurrence-free survival rates of 92.4% and 87.6% respectively and the median recurrence time of 15 (9 to 42) months. There were 25 recurrences in the thoracoscopy-laparoscopy group with 1- and 3-year recurrence-free survival rates of 81.7% and 67.9% respectively and the median recurrence time of 9 (3 to 42) months. There was significant difference in recurrence-free survival between the two groups (χ 2=4.193, P=0.041). Conclusions:The robotic surgical system has good oncology effect and surgical safety in the radical operation of esophageal cancer, which deserves further research and promotion.
		                        		
		                        		
		                        		
		                        	
4.Research updates on surgical treatments for portal hvpertension
Jinwei YANG ; Zhen MA ; Jike HU ; Tianliang SONG ; Xiaohong LIU ; Chunyu GENG ; Zhijian HAN ; Yumin LI ; Hao CHEN
Chinese Journal of Hepatobiliary Surgery 2017;23(9):640-645
		                        		
		                        			
		                        			Portal hypertension is a common clinical syndrome in chronic liver disease,such as schistosomiasis,portal vein occlusion cirrhosis and so on,which can be diagnosed when the hepatic venous pressure gradient is (HVPG) > 5 mmHg (1 mmHg =0.133 kPa).It could lead to gastroesophageal varicose veins rupture,ascites,spontaneous bacterial peritonitis,hepatorenal syndrome,hepatopulmonary syndrome,hepatic encephalopathy and some other serious complications,and is the primary death cause in cirrhosis and liver transplantation.The development of portal hypertension has experienced 4 phases ineluding the research about portal hypertension related theories and animal trial phase,preclinical tests and data accumulation phase,devascularization and shunts rapid development phase,the development phase of new technologies such as interventional and endoscopic surgical treatment,liver transplantation since the middle of the 19th century.The surgical procedures have been modified,which greatly reduce the complication and improve the life quality after operation.But so far none of them can cure portal hypertension thoroughly.This paper not only introduces the pathophysiologic basis of the surgical treatment,but also reviews the history of its development to summarize the recent progress,which may facilitate its surgical treatment.
		                        		
		                        		
		                        		
		                        	
5.Mechanism of bone marrow mesenchymal stem cells in promoting lung cancer metastasis
Tianliang ZHENG ; Song ZHAO ; Haizhou GUO ; Guanghui CUI ; Dawei LIN
Chinese Journal of Tissue Engineering Research 2016;20(19):2783-2788
		                        		
		                        			
		                        			BACKGROUND:So far the positive or negative effects of mesenchymal stem cel s on tumor growth and metastasis are under discussion. OBJECTIVE:To explore the mechanism of bone marrow mesenchymal cel s in promoting lung cancer metastasis. METHODS:Primary rat bone marrow mesenchymal stem cel s were obtained by direct adherence method of the whole bone marrow, and differential adherence combined with digestion control method was performed to purify cel s. Lung cancer cel lines were cultured, and the effects of bone marrow mesenchymal stem cel s on the migration, invasion and metastasis of lung cancer cel s were observed by scratch test, cel invasion and migration assays. Orthotopic lung cancer models were established in rats and bone marrow mesenchymal stem cel s were seeded onto the left lung of rats. Then, pathological changes of lung tissues were observed at 14 days after transplannation. RESULTS AND CONCLUSION:After the scratch test, the migration rate of lung cancer cel s became higher, and the scratches healed with time. And after cel transplantation, the number of migrated lung cancer cel s increased, as wel as the ability of lung cancer cel s penetrating the Matrigel was strengthened. Besides, fibrous connective tissues could be found around the lung cancer tissues, and necrosis with distinct boundary and large tumor nuclei;the metastatic tissues showed obvious infiltration and necrosis with large tumor nuclei. These results suggest that bone marrow mesenchymal stem cel s can promote the invasion, migration and metastasis of lung cancer cel lines.
		                        		
		                        		
		                        		
		                        	
6.Human umbilical cord mesenchymal stem cells:isolation, identificationand transplantation combined with chemotherapy for lung cancer in mice
Tianliang ZHENG ; Song ZHAO ; Haizhou GUO ; Guanghui CUI ; Dawei LIN
Chinese Journal of Tissue Engineering Research 2016;20(23):3457-3463
		                        		
		                        			
		                        			BACKGROUND:Mesenchymalstem cels have pluripotent differentiation, and can promote cel engraftment and immune regulation. Therefore,we attempt to use human umbilical cord mesenchymal stem cels as anew source for treatment of lung cancer by exploringcelisolation, identification and transplantation combined with chemotherapyforlung cancer in mice. 
 OBJECTIVE:To investigate the isolation and identification of human umbilical cord mesenchymal stem cels and its transplantation combined with chemotherapy for lung cancer inmice. 
 METHODS:Human umbilical cord mesenchymal stem cels were isolated from fresh umbilical cord of newborns and identified using tissue culture and enzyme digestion. Twenty Balb/C nude mouse models of lung cancer were randomly divided into two groups:mice in chemotherapy group were given chemotherapy, and those incombinedgroup given combination of chemotherapy with human umbilical cord mesenchymal stem cel transplantation. 
 RESULTS AND CONCLUSION:Compared with the chemotherapy group, the gastrointestinal tract was rosy and shiny, intestinal mucosa was smooth and complete, and tumor mass and blood indexes significantly decreased in thecombinedgroup (P< 0.05). To conclude, mature human umbilical cord mesenchymal stem cels can be obtained by tissueculture and enzyme digestion, andthecel transplantation combinedwith chemotherapy can significantly reduce gastrointestinal tract damage and themake peripheral hemogram in a stable level.
		                        		
		                        		
		                        		
		                        	
7.Establishment of dual liver transplantation rat model
Hao CHEN ; Tianliang SONG ; Zhijian HAN ; Aiqun ZHANG ; Xiaohong LIU ; Zhengkui PENG ; Yumin LI
Chinese Journal of Hepatobiliary Surgery 2016;22(9):630-632
		                        		
		                        			
		                        			Objective To establish a dual liver transplantation rat model,which could benefit the future clinical practice.Methods Y type vein derived from the crossover segment of vena cava and two iliac veins in donor and Y type bile duct prosthesis were employed to recanalize portal vein and bile duct from dual liver grafts to recipient liver.The dual right upper lobes with about 45% ~ 50% of the recipient liver volume were taken as donor.One was orthotopically implanted at its original position,while the other was rotated 180° sagittally and heterotopically positioned in the left upper quadrant.Survival rate was analyzed to evaluate the function of dual liver grafts.Results A total of 7 rats which underwent dual liver transplantation survived more than 7 days and the survival rate was 58.3%.5 rats died due to abdominal hemorrhage,bile leakage and liver abscess.Conclusion Using Y type vein and bile duct prosthesis,we successfully established a novel rat model of dual right upper liver lobe transplantation.
		                        		
		                        		
		                        		
		                        	
8.Expression of tumor necrosis factor receptor associated factor 6 in human esophageal carcinoma and its clinical significance
Tianliang ZHENG ; Song ZHAO ; Haizhou GUO ; Guanghui CUI
Chinese Journal of Primary Medicine and Pharmacy 2016;23(22):3437-3440
		                        		
		                        			
		                        			Objective To investigate the expression and clinical significance of tumor necrosis factor recep-tor associated factor 6(TRAF6)in human esophageal cancer.Methods The clinical data of 72 patients with esopha-geal cancer were collected.Immunohistochemistry method was used to determine TRAF6 expression in esophageal carcinoma and its adjacent normal tissue,and its relationship with clinical pathological features was explored.Results The TRAF6 positive expression rate in esophageal cancer tissue was 66.13%,which was significantly higher than that of normal tissue (13.89%),the difference between the two groups was statistically significant(χ2 =56.850,P <0.01).And TRAF6 expression level was significantly correlated with esophageal cancer clinical staging,lymph node metastasis(χ2 =6.818,4.428,all P <0.05),but TRAF6 expression was not correlated with age,sex,tumor differenti-ation.Conclusion The expression level of TRAF6 in esophageal carcinoma was significantly increased,and there was a significant correlation between the TRAF6 expression level and clinical pathological characteristics.
		                        		
		                        		
		                        		
		                        	
9.Radiation dose reduction to the lens of eye with bismuth shielding in head and neck MSCT
Yantao NIU ; Yaoyao SONG ; Yongxian ZHANG ; Senlin GUO ; Zongrui ZHANG ; Tianliang KANG ; Yunfu LIU
Chinese Journal of Radiological Medicine and Protection 2015;35(2):149-152
		                        		
		                        			
		                        			Objective To explore the effects on image quality and dose reduction to the lens when using bismuth shielding in head and neck MSCT.Methods The standard phantom and the cadaveric head with none,1,2 and 3 layers of bismuth shielding were scanned with protocols of brain,temporal bone and paranasal sinuses using a 16-MDCT scanner.The organ dose to the lens in each scanning was measured with thermoluminescence dosimeters(TLD).The above scanning with sinus protocol was repeated with a sponge with thickness of 5,10,15 and 20 mm placed between the shielding and phantom/head.The CT attenuation of phantom with the distance of 2,4,6 and 8 cm to shielding were measured.The image quality was subjectively evaluated by 2 physicians.Results The organ doses of the lens with protocols of brain,temporal bone and sinuses were 24.31,27.60 and 20.01 mGy,respectively.The doses were decreased significantly when using bismuth shielding.With the increase of the shield gap,the degree of dose reduction was reduced,but the increasing degree of CT attenuation significantly reduced.Using 2-layer and 3-layer bismuth shield in brain and temporal bone CT scan,the radiation doses were reduced by 47.1% and 59.1%,respectively while maintaining the diagnostic image quality.Using 1-layer shield without gap and 2-layer shield with 1.5 cm gap in sinus CT,the radiation doses were reduced by 31.5% and 34.5%,respectively.Conclusions Reasonable usage of bismuth shielding can effectively reduce the radiation dose to the lens of eye in head and neck MSCT.
		                        		
		                        		
		                        		
		                        	
10.Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis
Hao CHEN ; Zhijian HAN ; Zhengkui PENG ; Xiaohong LIU ; Tianliang SONG ; Shusen ZHENG ; Yumin LI
Chinese Journal of Organ Transplantation 2015;36(10):611-615
		                        		
		                        			
		                        			Objective To compare the cuffed renal vein technique and the classical techniques in kidney transplantation.Method The classical techniques of mouse renal transplantation required clamping both vena cava and aorta simultaneously and carried out suture anastomoses of the renal artery and vein in a heterotopic position.In our laboratory,we have successfully developed mouse orthotopic kidney transplantation for the first time,using a rapid cuffed renal vein technique for vessel anastomosis,wherein the donor's renal vein was inserted through an intravenous catheter,folded back and tied.During grafting,the cuffed renal vein was directly inserted into the recipient's renal vein without the need for clamping vena cava and suturing renal vein.Result This technique allowed for the exact transplantation of the kidney into the original position,compared to the classical technique,and had significantly shortened the clamping time due to a quicker and more precise anastomosis of renal vein as described.The renal vein anastomoses time was dramatically shortened in cuffed renal vein technique (4 min) as compared with the classical technique (9 min,P<0.001).This also allowed for a quicker recovery of the lower extremity activity,reduction in myoglobinuria with resultant kidney graft survival of 88.9%.Conclusion The cuffed renal vein technique simplifies microvascular anastomoses and affords important additional benefits.
		                        		
		                        		
		                        		
		                        	
            
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