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.
ObjectiveTo summarize the experiences and lessons of diagnosis and treatment of follicular thyroid carcinoma (FTC) with lymph node and lung metastases.MethodThe clinicopathologic data of a case of FTC with metastases of cervical, mediastinum, axillary lymph nodes, and bilateral lungs were analyzed retrospectively.ResultsThe case was a 39 years old male patient, who was diagnosed with the right FTC with multiple metastases. The total thyroidectomy+lymph node dissection in bilateral central and bilateral neck regions+lymph node dissection in the left axillary was intended to perform in the Department of Thyroid and Parathyroid Surgery of the West China Hospital. During the operation, the upper mediastinal lymph node fusion was found, and the tumor was tightly adhered with the recurrent laryngeal nerve, the signal was lost after the adhesion separation. In the right central area, the enlarged lymph nodes penetrated down into the upper mediastinum. More enlarged lymph nodes extended upward into the subclavian area in the left armpit. Considering a longer operation time and about 2 000 mL bleeding amount, the mediastinal and left neck operation was decided to perform in the second stage after consulting with the thoracic surgeon. At more than 6 months after the operation, the patient underwent the mediastinal lymph node dissection+superior vena cava (SVC) restoration+metastatic tumor resection+SVC shunt operation in the Department of Thoracic Surgery of the West China Hospital. The operation was successful, without hypocalcemia, dyspnea, and other complications. At 8 months after the operation, the third operation was performed in the Department of Thyroid and Parathyroid Surgery of the West China Hospital, that was, total residual thyroidectomy+left central and left cervical lymph node dissection+left axillary lymph node dissection (level Ⅱ–Ⅲ). The postoperative pronounce didn’t change, without hypocalcemia and other complications. Three times of iodine ablation therapy were performed in the West China Hospital. The occupy didn’t be found by the neck ultrasound and the pulmonary metastasis was stable by the CT during the regular follow-up.ConclusionsAccording to this case, neck surgery with SVC syndrome, SVC syndrome should be treated preferentially. For patient with complicated condition, unconventional approaches could be selected according to actual condition of patient, and staging surgery might be a good choice.
【摘要】 目的 分析成都市近年一线抗结核药的耐药状况，为耐药结核病预防控制提供依据。 方法 对成都市2007年1月-2009年12月就诊的结核患者，临床分离株培养鉴定为结核分枝杆菌的菌株采用绝对浓度法进行一线抗结核药:链霉素（SM）、异烟肼（INH）、利福平（RF）、乙胺丁醇（EMB）耐药性检测，分析结核分枝杆菌的耐药情况。 结果 1 235例结核患者中，总耐药率和总耐多药率分别为28.83%、14.01%，初始耐药率和获得性耐药率分别为12.82%、61.27%。近3年耐多药率有下降趋势，但获得性耐药率呈逐年上升趋势。 结论 成都市结核耐药状况仍然比较严重，进一步加强耐药结核的监测和控制非常重要。【Abstract】 Objective To analyze the drug resistant treating mycobacterium tuberculosis (MTB) in Chengdu in recent three years, and to provide the evidence for tuberculosis controlling. Methods The patients with MTB diagnosed from January 2007 to December 2009 in Chengdu were enrolled. Absolute concentration method was used to test the drug-resistance of streptomycin (SM), isoniazide (INH), rifampicin (RFP), and ethambutol (EMB). Results The total rate of drug resistance and multi-drug resistance were 28.83% and 14.01% respectively. The rates of initial drug resistance and the acquired drug resistance were 12.88% and 61.27% respectively. Multi-drug resistance rate showed a downward trend, but the rate of acquired drug resistance increased gradually. Conclusion The situation of drug resistance of tuberculosis in Chengdu is still serious, and it′s very important to further monitor and control the drug resistance treating tuberculosis.
【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 explore the predictive value of peripheral blood cells in the efficacy of neoadjuvant immunotherapy combined with chemotherapy for esophageal squamous cell carcinoma. Methods A retrospective study was conducted on patients with esophageal squamous cell carcinoma (clinical stages Ⅱ-Ⅳa) who underwent neoadjuvant immunotherapy combined with chemotherapy at the Department of Thoracic Surgery, Affiliated Hospital of North Sichuan Medical College from April 2020 to November 2023. According to whether the pathology was completely relieved after treatment, patients were divided into a pathological complete remission group and a pathological incomplete remission group. According to the CAP criteria for tumor pathological regression grading after neoadjuvant therapy, patients were divided into groups (TRG=0, 1 defined as good efficacy, TRG=2, 3 defined as poor efficacy). Results A total of 92 patients with esophageal squamous cell carcinoma were included, including 72 males and 20 females. The average age was 65.86±7.66 years. The complete remission of pathology was closely related to the number of lymphocytes in the blood before treatment (P=0.019). The AUC for predicting complete remission of esophageal squamous cell carcinoma after neoadjuvant immunotherapy combined with chemotherapy was 0.678, the maximum Yoden index was 0.328, and the optimal cutoff value was 1.845. The incidence of postoperative pulmonary infection in the group with incomplete pathological remission (25% vs. 5.6%, P=0.030) was higher than that in the group with complete pathological remission. According to the optimal cutoff value classification, there was a statistically significant difference (P<0.05) in the pathological N stage and pathological TNM stage between the two groups. The efficacy response was closely related to the number of red blood cells in the blood before treatment (P=0.009). The AUC for predicting TRG response after neoadjuvant immunotherapy combined with chemotherapy in esophageal squamous cell carcinoma was 0.669, the maximum Yoden index was 0.385, and the optimal cutoff value was 4.235. There were significant differences in postoperative pathological T staging (P=0.000), N staging (P=0.041), and TNM staging (P=0.000). According to the optimal cutoff value classification, there was a statistically significant difference in age (P=0.000) and hypertension (P=0.022) between the two groups. Conclusion Before neoadjuvant therapy, lymphocyte absolute value≥1.845 and red blood cell count<4.235 have good predictive effects on pathological complete remission and pathological response of esophageal squamous cell carcinoma after neoadjuvant immunotherapy combined with chemotherapy.
Objective To establish chronic hindlimb ischemia model with suture-occluded method in rats， and then compare the effects of chronic hindlimb ischemia model with acute ischemia model. Methods Models of chronic hindlimb ischemia were established by using suture-occluded femoral artery method. The laser Doppler blood flow analysis and angiography were performed on day 7， 14， 28， 42， and 49 after operation， and then the rats were sacrificed after angiography， respectively， the quadriceps and gastrocnemius of contralateral and ipsilateral （surgical side） were gotten， which were tested by HE staining and α-actin immunohistochemistry staining， and then calculate arteriolar density. Results There were no lameness and limb necrosis after operation in chronic hindlimb ischemia models. Laser Doppler analysis found that chronic hindlimb ischemia models were still maintained in ischemia state on day 49 after operation compared with acute ischemic models. The resluts of HE staining showed no acute necrosis and muscle fibrosis in chronic hindlimb ischemia model group. Chronic hindlimb ischemia models after operation did not appear obvious lameness and limb necrosis. The arteriolar density of quadriceps femoris on day 7 after operation in chronic hindlimb ischemia models were less than that in acute hindlimb ischemia models （0.015 2 vs. 0.036 4）. Conclusions Compared with the commonly used acute ischemic models， the duration of arterial limb ischemia in chronic hindlimb ischemia rats， which were established by suture-occluded method， is longer and less likely to be affected by the compensatory mechanisms. So suture-occluded method can provide a new animal hindlimb ischemia model for further study of ischemia angiogenesis mechanism and treatment of severe lower extremity ischemia.
Objective To evaluate effect of hypoxia condition (1% or 5% oxygen concentration) on proliferation, adhesion, migration, or viability ability of bone morrow-derived endothelial progenitor cells (EPCs). Methods The bone marrow mononuclear cells of SD rat were acquired with density gradient centrifugation method. They were cultured, induced, and differentiated to the EPCs. Then they were cultured respectively in three different oxygen concentrations (1%, 5%, or 21%). On the 3rd day and the 7th day, the effects of the different oxygen concentrations (1%, 5%, or 21%) on the EPCs’ neovascularization characteristics (including proliferation, adhesion, migration, and viability abilities) were evaluated. Results Whether cultured for the 3rd day or 7th day, the proliferation, adhesion, migration, and viability abilities of the cultured cells in the 1% and 5% oxygen concentrations were significantly better than those of the cultured cells in the 21% oxygen concentration (all P<0.05). Except for the proliferation ability of the cultured cells in the 5% oxygen concentration was significantly better than that of the cultured cells in the 1% oxygen concentration (P<0.05) on the 3rd day, and the adhesion ability on the 3rd day and the proliferation ability on the 7th day had no significantly differences, the other abilities (adhesion, migration, and viability abilities) of the cultured cells in the 1% oxygen concentration were significantly better than those of the cultured cells in the 5% oxygen concentration (allP<0.05). Conclusion Different oxygen concentration has an effect on proliferation, adhesion, migration, or viability ability of bone morrow-derived EPCs, appropriate hypoxia condition (1% or 5% oxygen concentration ) can enhance these abilities.