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Blockade of OX40/OX40L pathway promotes CD4 + CD25 + T regulatory cells proliferation

Pengji GAO ; Xisheng LENG ; Dong ZHANG ; Tao LI ; Jiye ZHU

Chinese Journal of General Surgery.2010;25(10):822-825. doi:10.3760/cma.j.issn.1007-631X.2010.10.013

Objective To evaluate the feasibility of OX40L gene silence in dentritic cells by RNAi through lentiviral vector, so that to explore the influence of CD4 + CD25 + T regulatory cells by blocking OX40/OX40L costimulation signals. Methods Serial plasmids were constructed, consisting of lentiviral vector framework, containing different OX40L siRNA sequences. The most effective packaged one by 293T cells to be the operating siRNA vector named OX4OL-RNAi-LV was chosen. The negative comparing vector was NC-GFP-LV. DCs isolated from C57BL/6 mice by magnetic selecting system were cultured in vitro and divided into 3 groups. Experimental group and negative control group were transfected with OX40L-RNAi-LV and NC-GFP-LV, respectively, and the blank control group was given the same volume culture solution. The MOI was 25. The status of transfection and GFP expression was monitored by GFP fluorescence. 6 days later, CD86 positive DCs were isolated and were co-cultured with CD4 + CD25 + T regulatory cells isolated from na(y)ve BALB/C. 6 days later, the proliferation and apoptosis of Tregs were evaluated by flow cytometry.Results The most effective siRNA sequence targeted the C,CTCATACAAGAATGAGTA episode of OX40L gene. The suppressive ratio of the OX40L protein expression was 73.1%. While the MOI was 25, the DCs transfected ratio by lentiviral vectors was at the level of 86. 4%. After co-cultured for 6 days, the CD4+CD25 +T regulatory cells of experimental group have a higher proliferation index (respectively, 38.3% vs.24.5 % ,22. 9%, F = 95.40, P = 0. 000) and lower apoptosis percentage ( respectively, 8.7 % vs. 20. 1%,19.8%, F=244.22,P=0. 000) than negative group and blank group. Conclusions The OX40L siRNA lentiviral vetor was constructed. This vector could effectively transfect DCs and block OX40/OX40L pathway, so to promote CD4 + CD25 + T regulatory cells proliferation in vitro.

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Pretransplantation HBV DNA load and post liver transplant HCC recurrence in HCC patients undergoing liver transplantation

Minru LI ; Shuhong YI ; Changjie CAI ; Guoying WANG ; Huimin YI ; Guihua CHEN

Chinese Journal of General Surgery.2010;25(9):717-719. doi:10.3760/cma.j.issn.1007-631X.2010.09.008

Objective To explore the relationship between hepatitis B virus (HBV) concentration and hepatocellular carcinoma (HCC) recurrence in HCC patients undergoing orthotopic liver transplantation (OLT). Methods 148 HCC patients associated with HBV infection undergoing OLT were enrolled in the study.Survival analysis was performed using the Kaplan-Meier method.Cox multiple regression analysis was performed to determine the parameters predicting HCC recurrence. Results Survival rates at 1,3 and 5 years were 86%,72% and 72%,respectively,and disease-free survival rate were 79%,71% and 54%,respectively.In this series 43 patients suffered from HCC recurrence.The recurrence rate was 29.1%(43/148),with the mean recurrence time being (13.16 ± 14.17) months (1 ～ 54 months).Exceeding Milan criteria (HR = 9.89; 95% CI 2.30 ～ 42.52; P = 0.002) and pretransplant HBV DNA level ＞ 5log10copies/ml (HR = 2.26; 95% CI 1.01 ～ 5.04; P = 0.047) were significant independent predictors for posttransplant HCC recurrence. Conclusion High HBV DNA load before transplantation is statistically associated with recurrence of HCC after liver transplantation.

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Changes of esophagogastric varicosis and portal hemodynamics in cirrhotic portal hypertensive patients after Hassab procedures

Yingjian ZHANG ; Jiansheng LI ; Wei WANG ; Dechun LIU ; Jianjun JIN ; Yufeng ZHENG

Chinese Journal of General Surgery.2010;25(9):706-709. doi:10.3760/cma.j.issn.1007-631X.2010.09.004

Objective To investigate changes of esophagogastric varicosis,and portal hemodynamics in cirrhotic portal hypertensive patients after Hassab surgical operation. Method The changes of esophageal varicosis and gastric varices incidences at different phases of cirrhotic patients after the Hassab surgical operation were examined by electric endoscope.Hemodynamics of portal vein system was measured by Doppler color imaging and color Doppler ultrasound after operation. Results Mild esophageal varicosis incidences on the 6th,12th and 18th month after the Hassab surgical operation were respectively 44.1%,34.7% and 28.4%,higher than that before operation (7.5%) (P＜0.05).Severe esophageal varicosis incidences on the 6th,12th and 18th month were respectively 25.1%,30.0% and 35.0%,lower than that before operation (65.2%) (P ＜ 0.05).Mild esophageal varicosis with gastric varices incidences on the 6th,12th,18th and 24th month before and after operation were respectively 16.6%,and 7.4%,9.2%,9.1%,10.3% respectively.Severe esophageal varicosis with gastric varices incidences in the 6th,12th,18th and 24th month were respectively 43.8%,and 45.8%,45.2%,47.4%,48.8% (P＜0.05).Inner diameter of portal vein and the right branch of portal vein on the 6th and 12th month before and after operation were respectively (13.5±1.7) mm,(9.8±2.0) mm,(10.5±2.2) mm,(11.2±1.2) mm,(8.1 ± 1.3) mm and (8.2 ± 2.2) mm (P ＜ 0.05).Maximum flow rate and blood flow of portal vein,velocity of the left of portal vein in the 6th and 12th month were respectively (13.6 ±2.6) cm/s,(1095 ±290) ml/min,(13.3±2.6) cm/s,(11.5 ±1.6) cm/s,(11.8 ±1.8) cm/s,(847 ±249) ml/min,(907 ± 310) ml/min,(11.0 ± 1.9) cm/s and 11.1 ± 1.9 cm/s (P ＜ 0.05).Velocity of the right of portal vein in the sixth month were (11.6 ± 2.6) cm/s,which was significantly slower than preoperative level of (13.2 ± 2.9) cm/s. Conclusion After the Hassab surgical operation the degree of esophageal varices lightens during 18 months and aggravates after 18 months.There is obvious relation between gastric varices and the degree of esophageal varices.Velocity of portal blood flow becomes slow,blood flow diminishes and inner diameter reduces after the Hassab surgical operation.

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Total laparoscopic splenectomy and pericardial devascularization for the treatment of portal hypertension

Xin DUAN ; Tao HE ; Zhi ZHENG ; Yingjian ZHENG ; Mingjie LI

Chinese Journal of General Surgery.2010;25(9):702-705. doi:10.3760/cma.j.issn.1007-631X.2010.09.002

Objective To evaluate total laparoscopic splenectomy combined with pericardial devascularization. Methods In this study,40 patients with portal hypertension and hypersplenism underwent total laparoscopic splenectomy combined with pericardial devascularization,in which 26 cases had a history of up-GI bleeding. Results Procedures were successful in 36 patients.There was four conversions to open surgery.The operations were completed in a mean time of (224 ± 54) min,with a mean estimated blood loss of (296 ± 209) ml and with no serious complication. Conclusion Total laparoscopic splenectomy combined with pericardial devascularization is a safe and effective available minimally invasive procedure in patients with portal hypertension and hypersplenism.

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Intraoperative internal carotid artery shunt in the resection of carotid body tumors (CBT)

Jichun ZHAO ; Yukui MA ; Bin HUANG ; Yi YANG ; Guojun ZENG

Chinese Journal of General Surgery.2010;25(7):533-535. doi:10.3760/cma.j.issn.1007-631X.2010.07.007

Objective To summarize the experience of surgical management of carotid tumors ( CBT ) and application of shunt between common and internal carotid artery intraoperatively. Methods Thirty patients of CBT (mean age:39. 2 ±2. 3 years old,10 male and 20 female, 15 in left, 14 in right and 1 in both sides) who underwent surgical resection, were retrospectively reviewed. The average size of CBT was 4.9 ±0.3 cm. The diagnosis was established by ultrasound, CT, MRI or carotid arteriography. 16 patients underwent surgical resection of CBT, 10 patients underwent additional ligation of external carotid artery, and 4 patients underwent additional intraoperative shunt between common and internal carotid artery. Results Surgical procedures were successfully performeded in all 30 patients with CBT. Intraoperative shunts were successfully used between common and internal carotid artery in 4 patients. The postoperative complications included hoarseness (15) , bucking (11), crooked tongue ( 17) , dyspnea (1), dysphagia(3). There was no hemiplegia and death. Conclusion Surgical resection is the choice of treatment of carotid body tumor. The application of intraoperative shunt between common and internal carotid artery in complicated Shamblin Ⅲ stage is safe and effective.

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Endovascular repair of acute standford type B aortic dissection complicated with massive hydrothorax

Chang SHU ; Mingyao LUO ; Quanming LI ; Ming LI ; Hao HE ; Xin LI

Chinese Journal of General Surgery.2010;25(7):529-532. doi:10.3760/cma.j.issn.1007-631X.2010.07.006

Objective To evaluate endoluminal repair and preoperative management for acute Standford type B aortic dissection complicating massive hydrothorax. Methods The clinical data of 27 patients (23 males, 4 females) hospitalized from January 2003 to December 2008 were analysed retrospectively. The average age was 47 ±9 years (35 ～70). Eleven patients had bilateral huge hydrothorax (40. 7% ) , while 13 had left hydrothorax (48. 1% ) and 3 had right hydrothorax (11. 1% ) only, and in 2 of them with additional pericardial effusion (7.4% ). SaO2 was below 90% in all cases. All patients underwent emergency endovascular repair. For control of massive hydrothorax found by CT or chest fluoroscopy, puncture drainage or tube drainage were administrated postoperatively. Results All the 27 operations were successful, and there was no perioperative mortality. Three type Ⅰ and one type Ⅳ endoleaks occurred but disappeared in one month. Hydrothorax disappeared 28 days to 3 months postoperatively in all patients, of which 5 cases had puncture drainage (18.5%) and 1 case had tube drainage ( 3. 7% ). Mean follow-up was 30 ± 20 m ( 6 ～ 78 m ) after endovascular management. Complications included pleural thickening (6 of 27, 22. 2% ) , pulmonary atelectasis (2 of 27, 7. 4% ) , and lung consolidation combined with chest dent (2 of 27, 7. 4% ). Conclusions Emergency endovascular therapy is safe and effective for acute Stanford type B aortic dissection with massive hydrothorax. Drainage of hydrothorax after stent-graft deployment is a must for the patient suffering from severe respiratory failure.

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Prophylaxis and treatment for complications associated with catheterization of brachial artery

Danming WU ; Yubin ZHOU ; Likui ZHANG

Chinese Journal of General Surgery.2010;25(7):526-528. doi:10.3760/cma.j.issn.1007-631X.2010.07.005

Objective To summarize our clinical experience of prophylaxis and treatment for complications associated with catheterization of brachial artery. Methods 87 patients underwent endovascular treatment via brachial artery. Complications associated with catheterization of brachial artery were retrospectively analyzed. Results Under ultrasonic guidance the catheterization procedure of brachial artery was successful in all 87 patients. The success rate of cannulation was 100%. In 53 patients(61% ) ultrasonic guided cannulation was successful at the first attempt, the other 35 (39% ) with 2 or more than 2 times puncture. Complications associated with catheterization of brachial artery were detected in 16 cases. The complication rate was 18. 4%. Guidewire insertion into peri vascular compartment took place in 4 cases (4. 6% ). Episodes of local hematomas were noted in 11 cases( 12. 6% ) and in three of the 11 cases nerve injuries were detected (one of which was of late-onset type). Brachial artery pseudoaneurysm was found in 1 case(1.2%). Conclusion Familiarity with the anatomical features of the brachial access, skillful application of the techniques for the catheterization, full understanding of complications and sufficient preparation of treatment strategies can prevent those complications effectively.

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Complications after carotid artery starting with the use of cerebral protective device

Daqiao GUO ; Xiao TANG ; Weiguo FU ; Junhao JIANG ; Bin CHEN ; Zhenyu SHI ; Lixin WANG ; Haolei CAI ; Yuqi WANG

Chinese Journal of General Surgery.2010;25(7):519-522. doi:10.3760/cma.j.issn.1007-631X.2010.07.003

Objective To review the prevention and treatment strategies for complications after carotid artery stenting in a single medical center. Methods From December 2004 to December 2009,clinical data of 118 cases of carotid artery stenting with the use of cerebral protective device were analyzed retrospectively. Results The surgical success rate was 100% with no perioperative 30- day mortality or severe complications such as disabling stroke. The overall perioperative complication rate was 32. 20% (38/118), including carotid artery spasm in 10.17% (10/118), bradycardia in 11.02% (13/118), hypotension in 2.54% (3/118), transient ischemic attack in 5.93% (7/118) and false aneurysm of femoral artery in 2. 54% (3/118) cases. The average follow-up time was (28 ± 15) months. The mortality was 2. 38% (2/84) and the overall complication rate was 10. 71% (9/84) , including ischemic stroke in 3. 57% (3/84) , in-stent restenosis in 1. 19% ( 1/84) , stent deformation in 4. 66% (4/84) and stent fracture in 1. 19% (1/84) cases. Conclusion Death or ischemic stroke and other severe complications after carotid artery stenting under cerebral protective device were low. A careful selection of patients and strict indication for surgery is the key to reduce the complication rate.

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Management of wrong coverage of carotid artery during endovascular repair of type Ⅲ aortic dissection by stented graft

Xiaoming ZHANG ; Xuemin ZHANG ; Chenyang SHEN ; Qingle LI ; Jingjun JIANG ; Yang JIAO ; Junlai ZHAO ; Tao ZHANG

Chinese Journal of General Surgery.2010;25(7):515-518. doi:10.3760/cma.j.issn.1007-631X.2010.07.002

Objective To evaluate the management of wrong coverage of the left common carotid artery ( CCA ) during interventional treatment of type Ⅲ aortic dissection by stented graft. Methods Coverage of the left CCA occurred in four cases during endovascular repair of type Ⅲ aortic dissection using stented graft. All four cases were male. The ages ranged from 37 to 45 years old with the average of 41. The first case was due to moving proximally of the first stented graft during delivery of one more cuff because of endoleak. Recanalization of the left CCA and subclavian artery ( SA) was acquired through sliding distally of the stented graft with the help of an aortic balloon. Coverage of the left CCA and SA in the second case was due to jumping-forward of stented graft during deploying, we dragged the stented graft distally through the pigtail catheter introduced from the left brachial artery with the help of a trap device. Then, the covered left CCA and SA were recanalized. The 2/3 left CCA was covered because of mislocation of the left CCA for the 3rd case. We recanalized the covered left CCA using chimney technique with the exposure of the left CCA. For the 4th case, half the innominate artery, the left CCA and SA were covered because the performer mistake the stented graft without naked stent as one with proximal naked stent. A bypass of assending aorta to bi-carotid arteries and the left axillary artery was performed using vascular graft one month after the endovascular repair. Result The proximal intimal entry was sealed successfully in all four cases. No endoleak, cerebral infarction and ischemia of the left upper limb occurred. Conclusion In cases of wrong coverage of carotid artery during endovascular repair of type Ⅲ aortic dissection, the CCA must be recanalized as soon as possible through surgical or interventional treatment in order to avoiding cerebral ischemia.

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Effects of HIF-1α expression regulated by Tet-on gone expression system on hepatoma cell proliferation and cell cycle in vitro

Ronglong XIA ; Zongquan XU ; Ruiqin ZHANG ; Xiaoping CHEN

Chinese Journal of General Surgery.2010;25(10):834-837. doi:10.3760/cma.j.issn.1007-631X.2010.10.016

Objective To investigate the effects of HIF-1α expression regulated by Tet-on gene expression system on cell proliferation and cell cycle of hepatoma cells in vitro. Methods The change of human hepatocellular carcinoma cell lines HepG2 cell cycle and cell proliferation was measured after HIF-1 α expression of HepG2 in vitro was regulated by Tet-on expression system. Results Amplified products were confirmed as the cDNA of HIF-1α by DNA sequencing, and pTRE-HIF-1α obtained by edonuclease digestion,capable of expression in HepG2 Tet-on cells. After being incubated under different concentrations of doxycycline for 48 h, MTT assays showed that up-regulation of HIF-1α expression increased HepG2 cell proliferation activities. The cell index of S and G2/M phase was significantly higher and that of G0/G1 phase reduced with the increasing concentrations of doxycycline. The mRNA expression of Cyclin A increased with the increasing concentrations of doxycycline ( P ＜ 0. 001 ), CyclinD1 and CyclinE did not change ( P ＞0. 05). Conclusion HIF-1 α gene promotes cell proliferation and cell cycle of hcpatoma cells in vitro, and this effects increased with the increasing of HIF-1α expression possibly through influencing the expression of CyclinA.

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