Standard venographies were pcrformed to evaluate the endothelial damage by the contrast medium. After different time intervals, the local veins were prepared for transmission and scanning electron microscopy (TEM and SEM) investigation. The veins were in a dilated state after the angiographies, which lasted for about two days. The endothelial damage was most severe 1 day after the venography. Besides the lesions of extensive endothelial tissurs, dcsquamations, and the exposure of subendothelial tissues, microthrombi somethimes were found. Healing occurred within 3 days. The results this study has also verifieed that it was more valuable to study venogqaphic effects on veins with TEM and SEM.
Objective To establish a scaffold model from heterogeneoussmall blood vessels. Methods Caudal arteries from 34 Wistar rats( average length 12.08±1.69 cm) were made into acellular blood vessel scaffolds. Some scaffoldswere observed by electron microscope, and others were transplanted to the cut ends of ear central arteries of male Japanese big ear white rabbits. Results Average external diameter was 0.74±0.08 mm in proximal, and 0.55±0.08 mm in distal end of rat caudal arteries. The small blood vessel scaffolds had shin wall whichwas white and soft, composed of fibrous tissues without cells. On the intima surface the fibrous tissues were arrayed densely in a grid-like pattern. After transplantation, the blood flow was reserved, and kept flowing freely in 24 hours. The pulsation of the transplanted artery was accessible and no blood leakage wasfound.Conclusion The natural scaffolds are composed of fibrous tissues, and can sustain the artery pulse pressure for 24 hours. It is better to suture the blood vessels by sleeve anastomosis.
Objective To summarize the methods, safety, and efficacy of the ex vivo liver resection followed by autotransplantation in the treatment of advanced hepatic alveolar echinococcosis (HAE). Method A retrospective analysis of clinical data and follow-up data in 21 cases who received ex vivo liver resection followed by autotransplantation in the treatment of HAE from February 2014 to December 2016 in West China Hospital was performed. Results All the patients successfully underwent ex vivo liver resection followed by autotransplantation and no death happened during operation. The median weight of remnant liver was 701.4 g (360–1 300 g), the average operation time were 13.6 h (9.4–19.5 h), the anhepatic phase time were 180–455 min with median of 314 min. The average of intraoperative blood loss were 2 379 mL (1 200–6 000 mL). The average of patients entered red blood cell suspension were 10.6 u (0–39.5 u), the average of fresh frozen plasma were 1 377 mL (0–6 050 mL) , of which 7 patients received autologous blood transfusion, with average of 1 578 mL (500–3 700 mL). The average of postoperative hospital stay were 23.5 days (4–51 days). Postoperative complications occurred in 12 patients during hospitalization, and 4 cases of postoperative complications were in grade Clavien-Dindo Ⅲ or above, 2 cases of grade Ⅴ (died). During the follow-up period, 19 patients were followed for a median of 16.2 months (3–38 months), no HAE recurrence or metastasis was found, only 1 patient were lost follow-up after surgery for 12 months. Massive ascites and hyponatremia were found in 1 patient who was diagnosis as left hepatic vein stenosis at the end of the 3 months after operation. The patient was cured after interventional treatment of hepatic vein stent implantation and angioplasty. Conclusions The ex vivo liver resection followed by autotransplantation provides radical treatment for patients with advanced HAE, but the surgery is difficult and has high risk of postoperative complications. The detailed preoperative evaluation, intraoperative pipeline reconstruction reasonably, and fine postoperative management can improve the patient’s survival, and reduce the rate of complications.
Objective To evaluate which is better method zymogen or low temperature frozen in removing vascular endothelial cell so as to lay a foundation for creating a kind of brace which is not to be rejected and the same as own blood vessel. Methods Fresh and not damaged umbilical blood vessel was collected from natural labour women, human umbilical blood vessel was remove carefully from normal foetus, then was put into disinfectant at 37℃ for 24 hours. They were divided into 3 groups:normal group(NG),zymogen group(ZG) and low temperature frozen group(LG). ZG: 0.1% collagenⅡ enzyme was addedin umbilical blood vessel and closed the both sides and the vascular endothelialcell was removed in 37℃ water. LG:Umbilical blood vessel was put into liquidnitrogen for 24 hours after frozened step by step, and then it was put into 37℃ water for 30-60 s and the vascular endothelial cells were washed away by normal saline. NG:Umbilical blood vessel was kept into 4℃ Kerb’s liquid. The bacteria were culturedin each group. The samples were stained by HE,elastic fiber and collagen fiberwere observed by light and scanning electron microscope. The difference of compliance was compared. Human leukocyte antigen ABC(HLA-ABC) and HLA-DR were observed by immunohistochemical method and the expression of antigen of umbilical blood vessel was analysed. Results In LG, umbilical vascular endothelial cells were removed completely; artery showed vertical smooth muscle and vein showed elastic membrane. InZG, umbilical vascular endothelial cells were removed completely after 20 minutes;artery showed vertical smooth muscle cells and vein showed lower endothelial layer. The vascular compliance in LG was higher than that in NG, and the latter was also higher than that in ZG,but showing no significant differences (Pgt;0.05). The compliance of umbilical vein was 2-3 times as much asthat of umbilical artery.The expression of HLA-ABC and HLA-DR in LG andZG were lower than that in NG, showing significant differences (Plt;0.01). Conclusion Low temperature frozen methodand zymogen method(0.1% collagen Ⅱ enzyme for 20 min) can remove vascular endothelial cells of human umbilical blood vessel completely.Low temperature frozenmethod was better than zymogen method.
Objective To study the surgical method to reduce bleeding in treating hemangioma at non-limb sites. Methods From November 1998 to November 2003,49 cases of non-limb hemangioma were treated, aged 3 months to 63 years, including 21 males and 28 females. There were 14 cases of capillary hemangioma, 25 cases of cavernous hemangioma, 7 cases of arterial racemose angioma and 3 cases of mixture hemangioma. According to the position and type of hemangioma, the various methods of blocking blood vessels were adopted to assist resect tumors. Afterthe pulsatile artery was felt in arterial racemose angioma of neck and face by palpation, we sutured and knotted it with 7-0 silk string to block the bleeding.We found out the common iliac artery or external iliac artery or femoral arteryand blocked them temporarily to resect arterial racemose angioma in inguen and thigh. We sutured and knotted vessel with 7-0 silk string to block the bleedingin capillary hemangioma and cavernous hemangioma of neck and face and truncus. Results Intraoperative bleeding obviously decreased and the tumor size reducedto various extent. Of the 49 cases, 47 cases achieved complete success, 2 casesbled within two days after operation. A postoperative follow-up of 6 months to4 years showed that the appearance and function were satisfactory. Conclusion The preoperative method of blocking blood vessels obviously can reduce intraoperative bleeding and decrease operative difficulty, which makes it possible to eradicate hemangioma and lower recurrence rate.
To study the feasibil ity of human adipose derived stem cells (ADSCs) in monolayer culture induced into smooth muscle cells in vitro as seeding cells in vascular tissue engineering. Methods The mononuclear cells in human adipose were separated by collagenase treatment and seeded on culture dishes with the density of 5 × 105/cm2. Cellswere cultured in M-199 plus 10% FBS. When reaching confluence, the cells were subcultured by 0.1% trypsin and 0.02%EDTA treatment, PDGF-BB (50 ng/mL) and TGF-β1 (5 ng/mL) were added at the passage 1 to enhance the smooth muscle cells’ phenotype. Cells were cultured under the inducing medium for 14 days. The morphology of induced cells was observed under the microscope. Cellular immunofluorescence and RT-PCR were used to determine the expression of smooth muscle cell markers of the post-induced cells. Flow cytometry (FACs) was used to examine the positive rate of induced team. Results Cocultured in M-199 media including TGF-β1 and PDGF-BB, the prol iferating capabil ity of the induced cells was significantly downregulated compared with the uninduced cells(P lt; 0.01). The induced cells exhibited “Hill and Valley” morphology, while the uninduced cells were similar to ADSCs of P0 which had the fibroblast-l ike morphology. The results of immunofluorescence indicated that the induced cells expressed smooth muscle (SM) cell- specific markers including α-smooth muscle actin (α-SMA), SM-myosin heavy chain (SM-MHC) and Calponin. The results of RT-PCR revealed that the induced cells also expressed α-SMA, SM-MHC, Calponin and SM-22α.The positive rates of α-SMA, SM-MHC and Calponin in FACs were 3.26% ± 1.31%, 3.55% ± 1.6% and 4.02% ± 1.81%, respectively, before the cells were induced. However, 14 days after the cell induction, the positive rates were 48.13% ± 8.31%, 45.33% ± 10.68% and 39.13% ± 9.42%, respectively. The positive rates in induced cells were remarkably higher than those in uninduced cells(P lt; 0.01). Conclusion The human ADSCs can be induced to express vascular smooth muscle markers, and they are a new potential source of vascular tissue engineering.
The experience on management of abnormal blood vessels in 128 cases of donor kidney during the tailoring operation was reported. The various techniques used for different types of abnormal arteries and veins, and the critical points which should be paid attention to have been discussed. It was concluded that the multiple renal arteries should be treated in a single renal artery and anastomosed with internal iliac artery or/and external iliac artery. The appropriate management given to abnormal renal blood vessels during the tailoring operation may shorten the warm ishemia time, ensure the renal blood supply, reduce the renal vasular complication, and promote the recovery of renal function.
PURPOSE:To observe tbe development of structure of the retinal blood vessels in human fetuses. METHODS:The retinas of 134 human fetuses from 12-weeks of fertilization to full term and of 4 adults were collected,and examined with immunohlstoebemical method. RESULTS:The examinations showed:(1)The spindle ceils only existed in the retinas of fetuseslt;27 weeks. Till full term,the vessels in ganglion cell layer(GCL)was still budding. (2)BM of all arterioles,venules and capillaries within the retinas of all fetuses and adults,was immunostained after exposure to antlsera against fibronectin.The laminin(LN)and type Ⅳ collagen(COL-Ⅳ)immunoreactive product appeared outside tbe endothelial cells of blood vessels in GCL at 25 weeks and 29 weeks,and tile b LN and COL-Ⅳ immunostaining around four layers of the retinal vessels in tile shape of sbeath appeared at 29 weeks and 32 weeks respectively. (3)As soon as the cords of the endothelial cells were canaized,the smooth muscle cells and perieytes appeared outside the walls. (4)While four layers of the retinal vasculature was spreading to the ora serrata,the glial limitans was formed outside the endothelial cells by the processes of astrocytes and Muuml;ller cells. CONCLUSIONS:Comparing with the adults,certain de fleieneies of the structure of the retinal vessels exisl in the human fetus,and tile endnthelial cells in the retina of the full lerm fetus arc still in proliferation and migration. （Chin J Ocul Fundus Dis,1997,13:153-156）
Since Oct. 1990, the 2nd metatarso-phalangeal joint and big-toe nail composite graft with the neuro-vascular bundle was transplanted to reconstruct the thumb in 4 cases. The transplants were all survived. The follow-up through 5 months, a comparatively good function and appearance were achieved.The applied anatomy, the surgical technique and the matters needing attention were detailed.
Objective To study the preparation method of acellular vascular matrix and to evaluate its biocompatibil ity and safety so as to afford an ideal scaffold for tissue engineered blood vessel. Methods Fresh caprine carotids (length, 50 mm) were harvested and treated with repeated frozen (—80 )/thawing (37℃), cold isostatic pressing (506 MPa, 4 ), and 0.125% sodium dodecyl sulfate separately for preparation of acellular vascular matrix. Fluorescence staining and DNA remain test were used to assess the cell extracting results. Biological characteristics were compared with the raw caprine carotids using HE staining, Masson staining, scanning electron microscope (SEM), and mechanical test. Biocompatibil ity wasdetected using cell adhesion test, MTT assay, and subcutaneously embedding test. Ten SD rats were divided into 2 groups (n=5). In experimental group, acellular vascular matrix preserved by the combination of repeated frozen/thawing, ultrahigh pressure treatment and chemical detergent was subcutaneously embedded; and in control group, acellular vascular matrix preserved only by repeated frozen/thawing and ultrahigh pressure treatment was subcutaneously embedded. Results HE staining and Masson staining revealed that no nucleus was detected in the acellular vascular matrix. SEM demonstrated that a lot of collagen fibers were preserved which were beneficial for cell adhesion. Fluorescence staining and DNA remain test showed that the cells were removed completely. There was no significant difference in stress and strain under the maximum load between before and after treatment. Mechanical test revealed that the acellular vascular matrix reserved mechanical properties of the raw caprine carotids. Cell adhesion test and MTT assay confirmed that cytotoxicity was grade 0-1, and the acellular vascular matrix had good compatibil ity to endothel ial cells. After subcutaneously embedding for 8 weeks, negl igible lymphocyte infiltration was observed in experimental group but obvious lymphocyte infiltration in control group. Conclusion The acellular vascular matrix, which is well-preserved by the combination of repeated frozen/thawing, ultrahigh pressure treatment, and chemical detergent, is an ideal scaffold for tissue engineered blood vessel.