Objective To investigate the effect of adenovirus vector mediated transfer of human herpes simplex virus thymidine kinase (HSVtk) gene inhibits intimal hyperplasia of vein grafts. Methods Auto vein graft models of Wistar rats were established. Adenovirus vector dwelled in cervical veins which were transplanted into inferior renal abdominal aorta. The combination of HSVtk (4×109 plaque forming units) and ganciclovir (GCV) was applied to test the inhibition effect. GCV was infused 〔60 mg/(kg·d), IP, Bid〕 from day 3 to day 21 after transplantation. Vein samples were harvested and the existence of HSVtk DNA was measured by PCR and the mRNA of it was studied by in situ hybridization. Van gieson (VG) and proliferating cell nuclear antigen (PCNA) stains were carried out in paraffin sections to study the thickness of neointima and smooth muscle cells (SMCs) proliferation with a computer-assisted analysis system. The apoptosis of SMCs also was detected by TUNEL. Results The existence of HSVtk gene in veins and its transcription were demonstrated. Morphometric analysis demonstrated a reduced intima thickness in the group receiving combination therapy (HSVtk/GCV) compared with HSVtk alone 〔(17.2±3.2) μm versus (31.1±2.5) μm, P＜0.05〕. GCV per se had no effect on intimal hyperplasia after vein transplantation. The apoptosis of SMCs increased significantly and expression of PCNA decreased in HSVtk/GCV gene therapy group versus blank control group 〔(9.1±2.3)% vs (28.7±3.6)%, P＜0.05; (38.7±5.6)%vs (18.5±2.6)%, P＜0.05〕. Conclusion GCV conditions reduction of intimal hyperplasia after intraluminal delivery of HSVtk in transplanting vena veins involving SMCs apoptosis.
Objective To explore the clinical features, treatment methods and prognostic factors of primary appendiceal tumors. Methods The clinicopathologic data of 82 patients with primary appendiceal tumor who met the inclusion and exclusion criteria and were treated in the Affiliated Hospital of Southwest Medical University from January 2013 to November 2022 were retrospectively collected, and the clinicopathologic characteristics of the patients were summarized. Log-rank and Cox proportional hazard regression models were further used for univariate and multivariate survival analysis for patients without other malignant tumors. Results Among the 82 patients with primary appendiceal tumor, 9 cases (11.0%) were complicated with other malignant tumors. Low-grade mucinous tumors were the most common (51.2%, 42/82), 25 cases were complicated with appendicitis and 12 cases were complicated with intestinal obstruction. Common clinical manifestations included abdominal pain, changes in bowel habits, abdominal distension, vomiting, nausea, etc. Among the patients without other malignancies, 69 patients were followed up for a median of 32 months (4–106 months), 18 of whom died. The 1-year, 2-year, 3-year and 5-year cumulative survival rates were 91.1%, 87.5%, 80.5% and 60.7%, respectively. Multivariate Cox proportional hazard regression analysis showed that platelet-to-lymphocyte ratio (P=0.004), anemia (P=0.015), tumor metastasis (P=0.007) and diabetes mellitus (P=0.011) were prognostic factors. Among them, patients with platelet-to-lymphocyte ratio >190.1, anemia, tumor metastasis and diabetes mellitus had poor prognosis. Conclusions The main pathological type of primary appendiceal tumors is low grade mucinous tumors. It lacks characteristic clinical manifestations, with abdominal pain and changes in bowel habits as the main manifestations, and can be accompanied by appendicitis and intestinal obstruction. There is no unified standard for treatment. For patients characterized by prognostic risk factors including platelet-to-lymphocyte ratio >190.1, anemia, tumor metastasis and diabetes, further monitoring and intervention are required.
Objective To assess clinical outcomes of therapeutic video-mediastinoscopy （VMS）. Methods Clinical data of 82 patients undergoing VMS in Zhongshan Hospital of Dalian University from December 2008 to October 2011 were retrospectively analyzed. Among them，24 patients received therapeutic VMS，including 18 men and 6 women with their median age of 56 （22-81） years. Three patients underwent operation through a neck incision，4 patients through a parasternal incision，and 17 patients through a lateral intercostal incision. Five patients received local anesthesia and basal anesthesia，and all the other patients received general anesthesia through single-lumen or double-lumen endotracheal intubation. Results Twelve patients with pleural effusion underwent pleural or lung biopsy and talc pleurodesis. Pathology examination showed malignant diseases in 11 patients and tuberculous pleural effusion in 1 patient. The median operation time was 35 （30-50） minutes，and postoperative hospital stay was 3-6 days. These patients were followed up for 1 month without recurrence of pleural effusion. Ten patients with mediastinal mass received pathological diagnosis and complete mass resection with their median operation time of 55 （30-270） minutes and median hospital stay of 7 （5-40） days. Two patients with hyperhidrosis underwent bilateral intercostal VMS sympathectomy. Their operation time was 60 minutes and 50 minutes respectively，and their hospital stay was 3 days. Postoperatively their sweating symptoms obviously resolved. They were followed up for 3 months，and their hands，feet and armpit were warm and dry. There was no in-hospital death in this group. Two patients （8.3%） had postoperative complications including 1 patient with phrenic nerve injury and another patient with pneumonia. Opioid analgesic drugs were not used postoperatively in 9 patients. Conclusion Therapeutic VMS is a safe，effective，minimally invasive and cosmetic procedure，but it is not suitable for resection of a large mediastinal mass.
Objective To investigate the effectiveness of percutaneous transluminal renal artery stenting (PTRAS) in treating atherosclerotic renal artery stenosis (ARAS). Methods A total of 69 patients with severe ARAS were treated with PTRAS between January 2002 and December 2008. There were 47 males and 22 females with an average age of 66.2 years(range, 42-88 years), including 66 cases of unilateral ARAS (single functional kidney, 1 case) and 3 cases of bilateral ARAS. Renal angiography revealed that the degree of renal artery stenosis was 70%-99%. Concomitant diseases included hypertension (67 cases), atherosclerosis obl iterans (69 cases), coronary heart disease (34 cases), diabetes (44 cases), and hyperl ipidemia (36 cases). Blood pressure, serum creatinine (sCr), and patency of the renal artery were measured to assess the effectiveness of PTRAS after 12 months. Results The renal artery stent was successfully implanted in 68 patients and the technical success rate was 98.6%. One patient was converted to il io-renal bypass because of intra-operative acute renal artery occlusion. One patient died of heart failure at 6 months after PTRAS, and 1 patient was lost at 3 months after PTRAS. The other 66 patients were followed up 32 months on average (range, 13-60 months). The blood pressure decreased significantly at 1 month and gained a further decrease at 12 months after PTRAS when compared with the preoperative ones [systol ic blood pressure: (132 ± 24) mm Hg vs (163 ± 34) mm Hg, P lt; 0.05; diastol ic blood pressure: (78 ± 11) mm Hg vs (89 ± 17) mm Hg, P lt; 0.05; 1 mm Hg=0.133 kPa]. Hypertension was cured in 4 cases (6.3%), improved in 52 cases (81.2%), failure in 8 cases (12.5%), and the overall benefit rate was87.5%. The sCr level was stable after 12 months of PTRAS, showing no significant difference when compared with preoperative basel ine [(107.8 ± 35.4) μmol/L vs (104.1 ± 33.8) μmol/L, P gt; 0.05]. Renal function was improved in 9 cases (13.6%), stable in 48 cases (72.8%), deterioration in 9 cases (13.6%), and the overall benefit rate was 86.4%. Instent restenosis found in 2 patients (3.0%) at 12 months after operation. Conclusion PTRAS is a safe and effective method to treat ARAS. It can control the blood pressure and stabil ize the renal function in most ARAS patients. Long-term efficacy needs further investigation.
Objective To observe the changes in the number and function of bone marrow-derived endothel ial progenitor cells (EPCs) after bone-marrow stimulation, and to investigate the possible mechanism of improving ischemicl imb disease after bone-marrow stimulation through autologue bone-marrow stem cell implantation. Methods Twelvemale Lewis rats, weighing 200-250 g, were classified into the bone marrow stimulation group (n=6) and the control group(n=6). In the stimulation group, the bone marrow of each rat was stimulated by injection of recombinant human granulocytemacrophage colony-stimulatory factor. Mononuclear cells were harvested from bone marrow and cultured in EBM-2 medium. After 7-day culture, EPCs were stained by 1, 1-dioctadecyl-3, 3, 3, 3-tetramethyl indocarbocyanine-labbled acetylated low density l ipoprotein/fluorescein isothiocyanate-ulex europaeus agglutinin 1, and the double positive cells were counted by the fluorescent microscope. The adhesive abil ity of EPCs was determined by counting the number of re-cultured EPCs. The unilateral ischemia hindl imb model was made with 12 Lewis rats. Three days later, EPCs were transplanted into the ischemic tissues. According to different sources of EPCs, the 12 rats were divided into 2 groups: the stimulation group (n=6) and the control group (n=6). At 3 weeks after EPCs transplantation, the quantity of the collateral vascular was observed by digital subtraction angiography (DSA). Results After 7-day culture, the number of EPCs in the stimulation and control groups was (145.2 ± 37.0)/HP and (95.2 ± 39.4)/HP, respectively, and there was significant difference between the two groups (P lt; 0.05). Meanwhile, the number of adhesive EPCs in the stimulation and control groups was (21.8 ± 4.3)/HP and (15.0 ± 5.2)/HP, respectively, and the difference between the two groups was significant (P lt; 0.05). At 3 weeks after the EPCs implantation, the number of the collateral vascular was significantly larger in the stimulation group (4.2 ± 1.2) compared with the control group (2.7 ± 0.8), (P lt; 0.05). Conclusion Bone marrow stimulation increases the number of EPCs and improves the function concurrently, which may be the reason why autologue bone-marrow stem cell implantation improves the curative effect of ischemic l imb diseases after bone-marrow stimulation.
【Abstract】ObjectiveSome studies have demonstrated that recombinant adenoviruses are efficient vectors for gene transfer to the venous wall and AdCMV.tk encoded thymidine kinase can be used to reduce restenosis. In this study AdCMV.tk was apply to human vein smooth muscle cells (SMC) and organ cultured saphenous veins to study its effects on proliferation of SMCs and reduction of intimal hyperplasia. MethodsThe adenovirus vector transferred tk gene and mark gene lacZ to the SMC of human saphenous veins and organ cultured vein segments. Various concentrations ganciclovir (GCV) were contained in culture media. The efficiency of gene transfer was studied by using Xgal staining. The proliferation of SMC was monitored by the method of trypan blue exclusion. The bystander effect was observed by mixed cell culture. After vein segments treated by AdCMV.tk+GCV and cultured for 14 days, HE and VG staining were carried out and intimal thickness was analysis by computer image system. ResultsAdenovirus vector could infect saphenous vein SMC efficiently both in cultured SMCs and organ cultured vein segments. Gene expression sustained 14 d at least. The inhibition of SMCs proliferation in vitro was a positive correlation in GCV concentrations and the levels of tk expression. The proliferation of SMCs transfectered lacZ wasn’t restrained by GCV (P＜0.05). In mixed cell experiment there was at least 55% reduction in total cell number when as few as 10% of the cells express tk. Assessment of this “suicide gene strategy” in saphenous vein organ culture model demonstrated that veins treated with AdCMV.tk+GCV had a significant reduction at 14 days in the intimal thickness compared to control group (P＜0.01). ConclusionThe results suggest that adenovirusmediated gene transfer of tk along with GCV administration may be a useful strategy to treat the proliferation of intimal hyperplasia of transplanting saphenous veins. Bystander effects are amplified by AdCMV.tk/GCV gene therapy system.
Objective To investigate the feasibil ity of preparing the porous extracellular matrix (ECM) by use of some chemicals and enzymes to decellularize the porcine carotid artery. Methods The porcine carotid artery was procured, and warm ischemia time was less than 30 minunts. The porcine carotid artery was decellularized with 1% sodium dodecyl sulfate (SDS) for 60 hours to prepare common ECM; then common ECM was treated with 0.25% trypsin (for 6 hours) and 0.3 U/ mL collagenase (for 24 hours) to prepare porous ECM. The common ECM and porous ECM were stained with HE,Masson’s trichrome, and Orcein to evaluate the histological features. Then the mechanical property, cytotoxicity, and pore size of ECMs were determined. After 4 weeks of subcutaneous implantation in dogs, the histological examination was used for the study. Results Histological observation confirmed that 2 kinds of ECMs were decellularized completely and more porous structure was observed in porous ECM. Scanning electron microscope showed the pores in porous ECM were greater and the length of shorter axis in porous ECM ranged from 5 to 30 μm, the length of longer axis from 40 to 100 μm. The porosity of porous ECM (99.25%) was greater than that of common ECM (91.50%). The burst pressure of porous ECM decreased when compared with common ECM, showing significant difference [(0.154 3 ± 0.012 7) MPa vs [0.305 2 ± 0.015 7) MPa, P lt; 0.05]. There was no significant difference in suture retention strength between 2 kinds of ECMs (P gt; 0.05). The cytotoxicity test showed no obvious cytotoxicity in 2 kinds of ECMs. In vivo implantation test showed that the deeper host cells infiltration and more neo-microvessels in porous ECM were observed than in common ECM. Conclusion SDS and some enzymes can be used to prepare porous ECM as the scaffold for tissue engineered blood vessels.
Objective To discuss the safety and feasibil ity of treating complex renal aneurysm with ex vivo aneurysmectomy and renal revascularization and renal autotransplantation after hand-assisted retroperitoneoscopic nephrectomy. Methods In October 2006, one male patient with complex renal aneurysm was treated. The preoperative color Doppler ultrasonograph, CT and DSA showed that there was an aneurysm (3.4 cm × 4.3 cm × 4.5 cm) located in the main renalartery bifurcation and its five branches of the left kidney. The patient had a history of hypertension with no response to treatment. After successful hand-assisted retroperitoneoscopic nephrectomy, the kidney off-body was perfused by the renal irrigating solution immediately to protect the kidney. Then ex vivo aneurysmectomy and renal artery revascularization were performed, the renal artery was reconstructed with an autologous right internal il iac artery. The reconstructed left kidney was re-implanted into the right il iac fossa. Results The operation was successful and the patient recovered without perioperative complications. The postoperative renal function was normal and the color Doppler ultrasonograph showed that the blood circulation in the transferred renal artery of the right il iac fossa and its branches was smooth, the blood circulation of the renal venous was smooth and no stenosis in the ureter 2 weeks after operation. Thirteen months follow-up showed the blood pressure was recovered to normal and the renal function was normal. Conclusion The method of ex vivo aneurysmectomy and autotransplantation is safe, feasible and minimally invasive for treating complex hilar renal artery aneurysms.
Objective To investigate the influencing factors for restenosis after femoral endarterectomy in treatment of arteriosclerosis obliterans at femoral artery . Methods A total of 103 patients with arteriosclerosis obliterans at femoral artery who underwent femoral endarterectomy from Jan. 2012 to Jan. 2017 in our hospital were retrospectively selected as subjects of this study, to compare the clinical feathers between restenosis group and patent group, and then exploring the influencing factors for restenosis after femoral endarterectomy. Results Thirty-six patients (35.0%) suffered from restenosis after femoral endarterectomy. Patients in the restenosis group had a high proportion of high smoking and diabetes mellitus, and high level of low density lipoprotein than those corresponding indexes of the patent group (P<0.05). Results of Cox proportional hazard model showed that, diabetes mellitus 〔RR=3.338, 95% CI was (1.003, 11.113), P=0.049〕 and high level of low density lipoprotein 〔RR=3.311, 95% CI was (1.166, 9.397), P=0.024〕 were independent risk factors for restenosis after femoral endarterectomy. Conclusions Monitoring of high-risk factors like controlling blood glucose strictly and strengthening statin treatment should be done to reduce the risk of restenosis after femoral endarterectomy for patients with arteriosclerosis obliterans at femoral artery.
Objective To explore the middle-term outcome of autologous bone marrow mononuclear cells transplantation in the treatment of lower l imb ischemia. Methods From March 2003 to June 2005, 65 patients with lower l imb ischemia were treated by autologous bone marrow mononuclear cells transplantation. Of the patients, there were 50 males and 15 females, with a mean age of 66.5 years (range 36-89 years), including 4 cases of simple arteriosclerotic occlusion,5 cases of thromboangiitis obl iterans and 56 cases diabetic lower l imb ischemia. A total of 400 mL bone-marrow blood were extracted from the posterior superior il iac crest. And then the mononuclear cells were isolated from the bone-marrow blood in the laboratory. The amount of transplantation bone marrow mononuclear cells was (0.60-1.80) × 109 (mean 1.05 × 109). Twelve patients received cell transplantation from two to four times and the other patients one time. According to the improvement of cl inical finding, the outcome was evaluated. Results All the patients were followed up for 8-56 months (mean 21.5 months). There were 8 deaths, and the mortal ity was 12.3%; 5 were due to myocardial infarction and heart failure and 3 were due to cerebral infarction. The general effective rate was 70.8% (46/65) and the recurrent rate was 10.7% (7/65). Of them, the response to treatment lasted over 12 months in 42 cases, accounting for 91.3% (42/46); over 24 months in 24 cases, accounting for 52.2% (24/46); and over 37 months in 12 cases, accounting for 26.1% (12/46). The effective rates were 100% in 12 patients who received 2-4 times transplantation and 64.2% in 53 patients who received 1 time transplantation, showing statistically significant difference between them (P lt; 0.001). Conclusion The middle-term outcome of autologous bone marrow mononuclear cells transplantation show that it is a feasible and simple method for treatment of lower l imb ischemia.