BACKGROUND:Endothelial progenitor cel s, known as the precursor cel s of mature endothelial cel s, have the function of neovascularization and neoendothelialization. Therefore, endothelial progenitor cel s have potential applicability in many fields. Endothelial progenitor cel s can be isolated and cultured from different resources with different methods, but the biological properties and identification of endothelial progenitor cel s stil have controversies.
OBJECTIVE:To explore the methods of isolation and culture of endothelial progenitor cel s from the human peripheral blood and to identify the biological features of endothelial progenitor cel s.
METHODS:Mononuclear cel s were isolated from the human peripheral blood using density gradient centrifugation, and the cel s were resuspended in endothelial basal medium-2 supplemented with the EGM-2-MV-SingleQuots. Then, the cel s were inoculated in human fibronectin-coated culture flasks and cultured in EBM-2MV medium. The morphology of endothelial progenitor cel s was observed. The proliferation potential and surface markers of endothelial progenitor cel s were characterized careful y. Furthermore, the functional properties such as nitric oxide release and tube formation on Matrigel were also evaluated.
RESULTS AND CONCLUSION:While adherent cel s maintained, spindle-shaped cel s formed a cel cluster after 6-7 days. Then, adherent cel s developed to endothelial progenitor cel s with a cobblestone appearance after 2-3 weeks. The endothelial progenitor cel s were confluent with an outgrowth appearance. Endothelial progenitor cel s had a higher proliferation potential compared with human aortic endothelial cel s under the same culture condition. Endothelial progenitor cel s expressed CD31, CD34, CD144 and KDR, displaying an obvious endothelial phenotype. Endothelial progenitor cel s were also found to uptake DiL-acLDL and exhibit lectin binding capability. Furthermore, endothelial progenitor cel s were able to form capil ary tubes on Matrigel and had the ability to release nitric oxide. Therefore, endothelial progenitor cel s can be obtained from the human peripheral blood by density gradient centrifugation and adherent culture. A combining method for the identification of endothelial progenitor cel s should be recommended.

Objective To discuss the experience in the diagnosis, treatment, complications and follow up of carotid body tumor. Methods All the 25 cases were diagnosised by DSA and CTA. The tumor was resected under carotid adventitial plane in 18 cases, with external carotid artery resection in 4cases, and in 3 cases, internal carotid artery (ICA) and external carotid artery (ECA) were resected simultaneously in which internal carotid artery was reconstructed in two cases including using self vein bypass in one and anastomosis between ICA and ECA in the other. ICA was ligated in the third case. Results No cases died perioperatively. ALL CBTs were treated successfully. Horner syndrome and trachyphonia were relieved after operation. Postoperative trachyphonia, bucking and lingual paralysis developed in 3 cases, and in one case with vagus resection caused dyspnea tracheotomy was performed. The rate of nerves injuries was 12% but no semiplegia and aphasia occurred. Follow up period was from 4 to 90 months (average 44 ±6 months) for 21 cases. The trachyphonia and bucking were improved during follow up but the lingual paralysis persists, and tumor recurred in two cases with one dying. Conclusions CBT treatment should include active surgery, sufficient preoperative preparation and avoiding the postoperative nervous complications.

BACKGROUND: At present, the commercial artificial small vascular grafts (diameter < 6 mm) are still unsatisfactory, due to poor blocompatibility and low long-term patency rate. Therefore, finding a vascular substitute with normal biological function and studying construction and function of tissue-engineered blood vessel have become hot topics recently.OBJECTIVE: To construct a novel tissue-engineered blood vessel by rabbit bone marrow mesenchymal stem cells (MSCs) and vascular acellular matrix, and to investigate the biocompatibility and patency rate of tissue-engineered blood vessels.DESIGN, TIME AND SETTING: An in vitro randomized controlled study at level of cytology and histopathology was performed at the Laboratory of Affiliated Drum Tower Hospital of Nanjing University Medical School from January 2006 to June 2008.MATERIALS: The decellularized vascular acellular matrix was obtained by a detergent-enzymatic procedure. MSCs from rabbits were isolated using density gradient centrifugation method and cultured in culture flasks coated with fibronectin. Subsequently, the expanded MSCs were seeded on the decellularized scaffolds, and then co-cultured in the self-made bioreactor to construct the tissue-engineered blood vessels.METHODS: Sixty rabbits were randomly divided into three groups. A1.0-cm abdominal aorta was sheared, and a tissue-engineered blood vessel was transplanted on the abdominal aorta using 8/0 polypropylene thread. Tissue-engineered blood vessel group: Tissue-engineered blood vessel was considered as the transplanted vessel; vascular acellular matrix group:Xenoma artery treated by vascular acellular matrix was considered as the transplanted vessel; xenoma artery group: Fresh xenoma artery was considered as the transplanted vessel.MAIN OUTCOME MEASURES: Immunocytochemical staining was used to identify the cultured MSCs. After 3 months of transplantation, the grafts were retrieved for digital subtraction angiography, pathological test and scanning electron microscope examination.RESULTS: Rabbits MSCs presented a whirlpool-like appearance at 8 days after culture. The immunocytochemistry results were consistent with the phenotype of MSCs. After high proliferation, MSCs were seeded onto the vascular acellular matrix for 12 days,and seed cells attached to well in the lumen of blood vessels. Three months after implantation, the patency rate was 90% of tissue-engineered blood vessel group and 80% of vascular acellular matrix group, which was superior to xenoma artery group (25%). At three months after transplantation, HE staining and scanning electron microscope demonstrated that internal, middle,and external membrane were clearly observed in the tissue-engineered blood vessel group, and the membrane morphology was similar to normal artery. The endothelial cells were covered completely. However, the endothelial cells were not covered completely in the vascular acellular matrix group, while mural thrombosis, mild proliferation of intima, and inflammatory cell infiltration were observed. The intima was thick and necrotic in the xenoma artery group, while lumens were stenotic and accompanied with a certain degree of thrombus organization.CONCLUSION: This study provides a new strategy to develop a tissue-engineered blood vessel with excellent biocompatibility and high patency rate constructed by rabbit MSCs and vascular acellular matrix.

Objective To investigate the technical details and therapeutic effect in treating StarIdford B aortic dissection by endovaseular aortic repair(EVAR). Methods We retrospectively review 115 type B aortic dissection cases undergoing the procedure of mini-traumatic EVAR from November 1998 to February 2009.There were 71 male patients and 44 female patients with a mean age of55.7 years and follow up period from 7 to 83 months for 75 cases. Results The procedure Was technically successful in 113 cases and 122 stent grafts were placed with hybrid surgery being carried out in 3 cases of carotid artery bypass.Severe perioperative complications included subclavian steal syndrome in 4 eases,pulmonaryinfections in 3 cases.arterial embolism in 2 cases,acute cerebral infarction in 2 cases,kidney tailure in 2 cases.alimentary tract hemorrhage,graft infection,acute intestinal ischemia and type A dissection occurred in one each case respectively.Five cases died perioperatively.Among the 75 cases that were fouowed up,there were 9 eases with type I endoleak and at last 5 cases redid endovaacular repmr.2 cases with secondary type A dissection accepted graft replacement.There were 6 deaths during the follow-up period,4 dying of acute myocardial infarction(2 cases),cerebral hemorrhage(1 case),type A dlssection (1 case)respectively,the causes of other2 deaths were not confirmed. Conclusion EVAR was effective for Standford B aortic dissection as shown by midterm follow up.

OBJECTIVESTo review the experience with the hybrid operations for patients with thoracoabdominal aortic diseases, as well as to evaluate and discuss the outcomes.

METHODSBetween June 2008 and September 2011, 11 patients (9 male and 2 female, mean age 52 years) were treated by hybrid operation. Hybrid operation includes debranching of the visceral arteries, followed by endovascular repair of the diseases (include aneurysmal disease, aortic dissection, or Takayasu's arteritis).

RESULTSAll the operations were successful. The mean operating time was 8.5 hours for patients done in one stage and 8.1 hours for the first stage of patient related in two stages. The endovascular stage took on average 2.0 hours. Overall mortality rate was 1/11 and morbidity was 4/11. No aneurysm-related death occurred. Two patients developed endoleak. Permanent paraplegia and renal failure rate was 0. Mean follow-up was 13.5 months.

CONCLUSIONHybrid operation is a mini invasive and safe method for thoracoabdominal aortic diseases with acceptable morbidity and mortality.

Adult ; Aged ; Aneurysm, Dissecting ; surgery ; Aortic Aneurysm, Thoracic ; surgery ; Aortic Diseases ; surgery ; Blood Vessel Prosthesis Implantation ; Female ; Humans ; Male ; Middle Aged ; Stents ; Vascular Surgical Procedures ; methods

Objective To verify the accuracy of multi-leave collimators (MLC) leaves position in intensity modulated radiation therapy (IMRT) using radiochromic films.Methods 7 medical electronic linear accelerators of Varian,Elekta or Siemens design were chosen.25 cm× 25 cm films were put on 30 cm×30 cm×3.0 cm homogeneous solid phantom and covered with a 2.0 cm thick of homogeneous solid phantom.The CT scanned images were transmitted to TPS for plan preparation.A 5 strip picket fence pattern was formed by MLC leaf,each 3 cm long and 0.6 cm wide,with 3.0 cm wide strip separation.At the maximum dose point,the SSD was 100 cm,with 250 MU per strip.After exposure,the films were transmitted to the dosimetry laboratory at IAEA for measurement and calculation.Results For 6 of 7 accelerators chosen,the differences of film-measured and TPS-planned MLC leaf position for every fence were within ± 0.5 mm as required by IAEA,and the other one not consistent with the requirements.The difference of film-measured MLC leaf position between each pair and all pair for 7 accelerators were within ± 0.5 mm,in line with IAEA's requirements.The differences of film-measured MLC leaf actual width were within 0.75 mm,as required by IAEA,for 6 accelerators and-0.8 mm for the other one,not consistent with the IAEA requirements.The standard deviations of film-measured MLC leaf actual width for all pairs for 6 accelerators were ＜0.3 mm,in line with IAEA requirements,but 0.4 mm for the other one,not consistent with IAEA requirements.Conclusions It is simple,fast and accurate to use radiochromic films for verification of the accuracy of MLC leaf position in IMRT.Therefore it is advisable to widely use radiochromic films in IMRT clinical practice.

OBJECTIVETo detect the expressions of miR-122 and miR-224 in hepatocellular carcinoma (HCC) by real-time fluorescence quantitative RT-PCR and investigate the significance of miRNAs in early diagnosis of HCC.

METHOD2(-Delta Delta CT) method was used for quantitative analysis of the expression pattern of miR-122 and miR-224 in 35 HCC and adjacent normal tissues. All the quantitative results were confirmed by Northern blotting.

RESULTSCompared with adjacent normal tissues, the HCC tissues showed significant miR-122 down-regulation (P<0.01) and miR-224 over-expression (P<0.001).

CONCLUSIONSHCC has obvious alteration in the expression patterns of miR-122 and miR-224, and real-time fluorescence quantitative RT-PCR provide a new means for early, efficient, and accurate diagnosis of HCC.

Adult ; Aged ; Base Sequence ; Blotting, Northern ; Carcinoma, Hepatocellular ; diagnosis ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; diagnosis ; genetics ; Male ; MicroRNAs ; genetics ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Spectrometry, Fluorescence ; Time Factors

ABSTARCT Purpose To investigate the clinicopathologic characteristics and genetic mutations of metastatic follicular thy-roid carcinoma(FTC).Methods A total of 22 cases of meta-static FTC were collected,including previous medical history,imaging,treatments and outcomes,and next-generation sequen-cing study and Sanger sequencing were performed in 12 cases.Results There were 16 women and 6 men.Sixteen cases were older than 50 years.Seven cases presented with metastases as the first symptom.Fourteen cases developed metastases 3 to 12 years after thyroid surgery.Sixteen cases developed bone metas-tasis,10 cases had lung metastasis,and 3 cases had brain me-tastasis.Those patients with multiple bone metastases progressed during the follow-up period.The common gene mutations in me-tastases were NRAS p.Q61R(6 cases),HRAS p.Q61R(2 ca-ses)and KRAS p.Q61R(1 case),followed by TERT promoter mutation(8 cases).Other mutated genes included KEL,BRCA1/2,ALK,ROS1,ErbB4,etc.Conclusion FTC has a high misdiagnosis rate.Those diagnosed with FTC should under-go regular systemic examinations to detect potential metastasis,especially in bone,lung,and brain.Further research on the sig-nificance of NRAS and other molecular indicators in FTC metas-tasis will help to better predict its biological behaviors.

Humans ; Lithiasis ; surgery ; Liver Diseases ; surgery ; Liver Transplantation ; Male ; Middle Aged ; Situs Inversus ; complications ; surgery

OBJECTIVETo evaluate the feasibility and efficacy of hybrid procedure for the treatment of multi-level iliac and common femoral occlusive disease.

METHODSFrom January 2008 to June 2011, 36 lower limbs with sever iliac and common femoral occlusive diseases were treated by iliac artery stenting combined with open femoral endarterectomy. The mean age of the whole study population was 65 years (range 49 to 87 years) with a male predominance (26 males, 72.2%). The early clinical results were determined by ankle brachial index and intermittent claudication distance. Patency analyses were performed using Kaplan-Meier life tables. Univariate and multivariate analysis were used to assess the influence of various risk factors on primary patency.

RESULTSAll lower limbs underwent successful hybrid surgical and endovascular therapy. Clinical improvement was seen in 94.4% of patients. The mean duration of follow-up was 24.2 months, overall, the primary patency rates, primary assisted patency rates and second patency rates were 72.2%, 83.3% and 94.4% respectively. The primary patency rate for intermittent claudication was significantly higher than that for critical limb ischemia (P = 0.041, 0.012). Cox regression analysis did not reveal any independent predictor of primary patency.

CONCLUSIONHybrid procedures provided an effective treatment management of multilevel iliac-femoral arterial occlusive disease.

Aged ; Aged, 80 and over ; Angioplasty, Balloon ; Arterial Occlusive Diseases ; surgery ; Endarterectomy ; Feasibility Studies ; Female ; Femoral Artery ; surgery ; Follow-Up Studies ; Humans ; Iliac Artery ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome