ObjectiveTo explore the antitumor effect of tumor vaccine fused from dendritic cells (DC) and Walker-256 cancer cells on implanted liver cancer in rats and the related mechanism of inhibition for tumor angiogenesis. MethodsWalker-256 cancer cells and mature DC were fused by 50% polyethylene glycol method for preparation of DC-Walker-256 fusion vaccines. Implanted liver cancer models were established through operations on healthy male SD rats at the age of 6－8 weeks. All the rats were divided into four groups, and rats in each group were injected subcutanely with fusion vaccine (group), mixed cultured cells (group), simple DC (group), and PBS (blank control group), respectively. On 28 d after making model, the rats were put to death, the tumor was observed and pathological essays were prepared. All rats’ spleens were collected and prepared into lymphocyte to detect antigenic specificity cytotoxic T lymphocyte (CTL) by enzymelinked immunosorbent spot (ELISPOT) method. The expressions of VEGF, ANG-1, ANG-2, and MVD were detected by immunohistochemistry. ResultsThe numbers of rats survived in the fusion vaccine group, mixed culture cells group, simple DC group, and blank control group was 8, 5, 6, and 3, respectively. The rats in the other three groups except for fusion vaccine group were manifested as inaction, anorexia, and gloomy fur in some degree as well as ascites. The tumorigenesis was found in all survival rats except for two in the fusion vaccine group. The weight of liver tumors of rats in the fusion vaccine group 〔(32.4±9.2) g〕 was significantly lighter than that in the mixed culture cells group 〔(67.3±5.1) g, P=0.031〕, simple DC group 〔(75.0±8.3) g, P=0.019〕, and blank control group 〔(86.6±10.5) g, P=0.008〕, respectively. The number of tumorspecific CTL of rats in the fusion vaccine group was also significantly higher than that in the other three groups (P=0.019, P=0.025, and P=0.001, respectively). The MVD of tumor tissue in the fusion vaccine group was (24.12±2.32) vessels/HP, which was significantly lower than that in the mixed culture cells group 〔(40.34±1.29) vessels/HP, P=0.025〕, simple DC group 〔(42.36±3.16) vessels/HP, P=0.035〕, and blank control group 〔(56.48±5.16) vessels/HP, P=0.006〕, respectively. The MVD of tumor tissue in the mixed cultured cells group and simple DC group was similar (P=0.165), however, which was significantly lower than that in the blank control group (P=0.040 and P=0.043). The positive rate of VEGFA protein expression was 23.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (42.5%, P=0.031), simple DC group (61.3%, P=0.019), and blank control group (89.6%, P=0.003), respectively. The positive rate of VEGF-A protein expression in the mixed cultured cells and simple DC groups was similar (P=0.089), however, which was significantly lower than that in the blank control group (P=0.027 and P=0.038). The positive rate of ANG-1 protein expression in the fusion vaccine group (43.2%) was not different from that in the mixed culture cells group (46.3%, P=0.292), simple DC group (51.3%, P=0.183), or blank control group (49.6%, P=0.179), respectively, and the difference of pairwise comparison in latter three groups was not significant (P=0.242, P=0.347, and P=0.182). The positive rate of ANG2 protein expression was 19.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (62.3%, P=0.007), simple DC group (67.3%, P=0.005), and blank control group (71.6%, P=0.004), respectively, however, the difference of pairwise comparison in latter three groups was not significant (P=0.634, P=0.483, and P=0.379). ConclusionFused vaccine can induce CD8+ CTL aiming at tumor cells and establish the effective antitumor immunity in vivo and also downregulate the level of VEGF and ANG-2 to suppress tumor angiogenesis and thereby achieve the purpose of curing tumor.
ObjectiveTo review the research status in respect of interspinous distraction devices (IDD) in the treatment of degenerative lumbar spinal stenosis (DLSS). MethodsRecent original articles related to IDD in the treatment of DLSS were retrieved extensively, and the effectiveness was analyzed and summarized. ResultsIts short-term effectiveness was superior to that of conservative treatment, no significant difference was found when compared with decompression or fusion alone. Its complication rate was higher than that of decompression or fusion alone, although complication dose not significantly affect treatment results, it still will increase the medical care cost of patients. ConclusionIDD is appropriate for patients who have failed to nonoperative treatment measures and can not tolerate open surgery, but the long-term effectiveness and durability of IDD need further research.
Objective To review the osteogenic mechanism and osteogenic effects of bone morphogenetic protein 6 (BMP-6) so as to provide the basis for further research of BMP-6. Methods The related articles about the osteogenic mechanism and the osteogenic effects of BMP-6 in experimental animals were extensively summarized. Results BMP-6 from bone matrix can transduct the osteogenic signal to bone marrow mesenchymal stem cells (BMSCs) by means of Smad protein signal transduction pathway. And the BMSCs which received the signals will differentiate into osteoblasts and chondroblasts. Therefore, BMP-6 plays an important role in the development and maturation of bone and cartilage. In addition, BMP-6 has a close relation with bone diseases, such as fracture, osteoporosis, and bone tumor. Conclusion The deep research of BMP-6 is expected to provide a new therapeutic approach for treating bone diseases of nonunion, osteoarthritis, and osteoporosis.
ObjectiveTo observe and compare the cytological and biological differences between human normal and degenerated nucleus pulposus (NP), and to investigate the repair effect of insulin-like growth factor 1 (IFG-1) and platelet derived growth factor (PDGF) on human degenerated NP.MethodsHuman degenerative and normal NP tissues were obtained from operative patients, a portion of which were processed into tissue sections and HE staining was performed to observe the morphological changes of nucleus pulposus cells (NPCs) before and after degeneration of NP. Immunohistochemistry staining was used to determine the expression levels of collagen type Ⅰ, collagen type Ⅱ, B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X (Bax) proteins. Another portion of tissues were isolated and cultured and NPCs morphology was observed under inverted microscope. Western blot analysis was used to detect collagen type Ⅱ protein expression. Then, the gene transfection experiments were launched, including 4 groups, with group A designed as degenerated NPCs only, and groups B, C, and D of degenerated NPCs transfected with IGF-1 gene lentiviral particles, PDGF gene lentiviral particles, and lentiviral particles carrying IGF-1 and PDGF double genes, respectively. At 21 days after transfection, the cell morphology of each group was observed under inverted microscope, the positive rates of IGF-1 and PDGF of each group were measured by flow cytometry, and the expression of collagen type Ⅱ protein was detected by using immunohistochemistry staining and Western blot.ResultsHE staining showed that there were a large number of notochordal cells and a small number of chondrocytes in the central NP tissue of normal group, while the NPCs in degeneration group were significantly reduced, and a large proportion of fibrocartilage tissues were found in NP tissue. Immunohistochemistry staining showed that the percentages of collagen type Ⅰ and Bax protein-positive cells in degeneration group were significantly higher than those of normal group, while the percentages of collagen type Ⅱ and Bcl-2 protein-positive cells were significantly lower than those of normal group (P<0.05). Western blot showed that the relative expression level of collagen type Ⅱ protein in degeneration group was significantly lower than that in normal group (t=65.493, P=0.000). At 21 days after gene transfection, compared with group A, the cell viability of groups B, C, and D increased and the morphology became more regular. Flow cytometry showed that the percentages of IGF-1-positive cells in groups B and D were significantly higher than that in group A, and the percentages of PDGF-positive cells in groups C and D were significantly higher than that in group A (P<0.05). Immunohistochemistry staining showed that the positive stainings of collagen type Ⅱ in groups A, B, C, and D was (±), (+), (+), and (++), respectively. Western blot showed that the relative expression of collagen type Ⅱ protein in groups A, B, C, and D increased by degrees, and the differences between groups were significant (P<0.05).ConclusionBoth IGF-1 and PDGF can reverse the degeneration of intervertebral discs NPCs and they have synergistic effects, providing experimental basis for its application in clinical treatment approaches for degenerative disc disease.
ObjectiveTo investigate the effectiveness of one-stage arthroscopic reconstruction and strict immobilization for 6 weeks for treatment of knee dislocation. MethodBetween August 2010 and May 2013, 22 cases (22 knees) of knee dislocation were treated with one-stage reconstruction and strict immobilization for 6 weeks. There were 15 males and 7 females, aged 21-54 years (mean, 31.5 years). The left knee and right knee were involved in 8 cases and 14 cases respectively. The disease causes were traffic accident in 12 cases, falling from height in 6 cases, and sports injury in 4 cases. The time between injury and operation was less than 2 weeks in 6 cases, 2-3 weeks in 10 cases, and more than 3 weeks in 6 cases. The results of anterior drawer test, posterior drawer test, and Lachman test were positive in all patients. The posterior displacement of the tibia was more than 10 mm. The results of valgus stress test and varus stress test were positive in 13 cases and 11 cases respectively. The preoperative knee range of motion was (58.2±28.4) °, Lysholm score was 39.7±4.6. All patients had anterior cruciate ligament rupture and posterior cruciate ligament rupture; combined injuries included medial collateral ligament rupture in 11 cases, lateral collateral ligament rupture in 9 cases, both medial and lateral collateral ligament rupture in 2 cases, femoral condylar avulsion fracture in 2 cases, and meniscus injury in 7 cases. No nerve or blood vessel injury was observed. ResultsAll cases obtained primary healing of incision without infection. All the patients were followed up 12-48 months (mean, 27.8 months). At 12 months after operation, the results of the anterior drawer test, posterior drawer test, Lachman test, valgus stress test, and varus stress test were all negative; the knee range of motion increased was significantly to (121.3±7.9) °(t=30.061, P=0.000) ; Lysholm score was 87.2±6.1, showing significant difference when compared with preoperative score (t=24.642, P=0.000) . ConclusionsA combination of arthroscopic one-stage reconstruction and strict immobilization for treatment of knee dislocation is a safe and effective method, good stability and joint function can be achieved.
ObjectiveIn order to correct left pulmonary artery (LPA) stenosis in tetralogy of Fallot (TOF), a technique for LPA reconstruction by widening pulmonary arterial posterior wall was devised. To evaluate its surgical technique as well as postoperative outcome. MethodsFrom January 2008 through August 2014 year, 1 142 consecutive patients underwent repair of TOF including 44 patients with TOF and LPA stenosis in our hospital. We used widening pulmonary arterial posterior wall for LPA reconstruction in 21 patients. The median age was 22.8 months (range, 1 month to 11 years), and the median weight was 9.1 kg (range, 3.8-29 kg). ResultsThere was one operative death. No death occurred during the follow-up period in the other 20 patients. There were 4 patients with complicate unbalanced pulmonary perfusion postoperatively. Echocardiography at 3-72 months follow-up demonstrated no obvious stenosis was found at LPA in 14 patients, and mild stenosis (32.8±12.7 mm Hg) in 6 patients. Only 1 patient required further interventions because of restenosis of LPA. ConclusionsLPA reconstruction by widening pulmonary arterial posterior wall is an effective method in the management of stenosis of LPA in the patients with TOF. We emphasize division of the arterial duct or ligamentum completely because it will tether and kink the LPA. This procedure retains the better growth potential by increasing autogenous tissue of native tissue.
Objective To investigate and compare the different surgical strategies for typeⅠpulmonary atresia with ventricular septum defect (PA/VSD) and the outcomes of postoperative prognosis in early stage. Methods We retrospectively analyzed the clinical data of 61 typeⅠPA/VSD patients (40 males, 21 females) with a median age of 249 days (range, 13 days-19 years) in Guangdong Cardiovascular Institute from January 2005 to December 2014 . Among them, 42 patients (27 males, 15 females) with a median age of 11.11 months, ranging from 0.80–211.70 months received radical surgery as a radical surgery group. And 19 patients (13 males, 6 females) with a median age of 2.96 months, ranging from 0.47–161.83 months underwent palliative surgery as a palliative surgery group. We compared the two surgeries and their early outcomes. Results The mean postoperative oxygen saturation was 88.08%±9.64%, which showed significant improvement compared with preoperative oxygen saturation of 74.08%±12.99% (P＜0.05). Patients in the palliative surgery group had a lower body temperature during cardiopulmonary bypass and more respiratory complications than those in the radical surgery group (24.69 °C±3.11 °C vs. 27.18 °C±2.10 °C). Conclusion Both radical and palliative surgeries are good for the increase of pulmonary blood volume and the development of pulmonary vessels. Surgeons must pay more attention to choosing radical surgery for the babies, which is only considerd for those with well developed pulmonary arteries.
Objective To observe the effect of Melittin on collagen type II (Col-II) expression of rat endplate chondrocytes (EPCs) induced by interleukin 1β (IL-1β). Methods Primary EPCs from the lumbar vertebra of 4-week-old Sprague Dawley rats were culturedin vitro and identified by morphological observation, toluidine blue staining and Col-II immunofluorescence staining. Then, MTT assay was used to determine the optimal concentration of IL-1 and Melittin. Next, EPCs at passage 3 were randomly divided into 4 groups: no treatment was done in group A as control group; the optimal concentration of IL-1β, Melittin, and both IL-1β and Melittin were used in groups B, C, and D respectively. The expression of Col-II was detected by Western blot after 48 hours intervention. Results Under inverted microscope, the first generation EPCs were polygonal; cell proliferation decreased after fifth generation, and cell morphology changed into fusiform. The acidic mucosubstance in the cytoplasm (such as Aggrecan) was stained dark blue by toluidine blue. After marking Col-II by immunofluorescence, the positive expression of cytoskeleton (green fluorescence) could be observed. MTT assay showed that IL-1β and Melittin could inhibit the EPCs in a dose-dependent manner after intervention of 24 and 48 hours, and the optimal concentrations of IL-1β and Melittin intervention were 10 ng/mL and 1.0 μg/mL respectively. Compared with group A, the expression of Col-II was significantly reduced in group B, and was significantly increased in group C by Western blot assay, but there was no significant difference between group D and group A. The Col-II expression levels of groups A, B, C, and D were 0.991±0.024, 0.474±0.127, 1.913±0.350, and 1.159±0.297 respectively, showing significant difference between the other groups (P<0.05) except between group A and group D (P>0.05). Conclusion Melittin has a protective effect on endplate cartilage, and the research results provide experimental basis for the prevention and treatment of spinal degenerative disease.
Objective To review the clinical experience of one-stage repair of interrupted aortic arch(IAA) in neonate with cardiac anomalies. Methods We retrospectively analyzed the clinical data of 21 patients (18 males, 3 females) with IAA total repair in our hospital between May 2003 and September 2014. The average age of patients was 6–26 (15.9±5.8) days and the mean body weight was 3.3±0.4 kg. Fourteen patients belonged to IAA type A, and 7 patients to type B. All patients were complicated with ventricular septal defect, atrial septal defect, and patent ductus artefious. All patients with cardiac anomalies underwent one-stage repair through median sternotomy. The aortic continuity was reestablished by anastomosis between the descending aortic segment and aortic arch. Results CPB time was 92–174 (132.6±27.1) min, and aortic cross clamping time was 48-118 (70.9±18.8) min. Hospital day was 4-52 (28.0±12.1) d. There were 3 postoperative deaths. Causes of death included a cardiac arrest in one patient, hematosepsis in one patient, and a pulmonary hypertension crisis in one patient. Eighteen patients were followed up for 3 months to 11 years and the results were excellent. Conclusion One-stage repair of IAA in neonate with cardiac anomalies can improve life quality of patients and achieve good results.
Objective To analyze the clinical efficacy and mid-term outcomes of reimplantation of anomalous origin of left coronary artery from the pulmonary artery (ALCAPA), and to evaluate whether concomitant management of mitral regurgitation (MR) during ALCAPA repair is needed. Methods Between March 2005 and March 2015, 52 consecutive patients (20 males and 32 females with a median age of 10 months ranging 2-193 months) underwent reimplantation of ALCAPA at Department of Cardiac Surgery, Guangdong Cardiovascular Institute. There were 14 males and 21 females with a mean age of 35.4±42.8 months not receiving mitral valvuloplasty (a Non-MVP group), and 6 males and 11 females with a mean age of 13.5±11.0 months receiving mitral valvuloplasty (a MVP group). In order to facilitate the analysis, degree of MR was graded by number: 0.0=none, 1.0=trivial, 2.0=mild, 2.5=mild-moderate, 3.0=moderate, 3.5=moderate-severe and 4.0=severe. Results The left ventricular fractional shortening (LVFS) and left ventricular end diastolic diameter (LVEDD) demonstrated significant improvement between preoperation and discharge (28.6%±9.6% vs. 32.1%±10.1%, P=0.023; 38.4±5.6 mm vs. 30.5±5.7 mm, P<0.001), and there was also significant improvement in the mean MR grade between preoperation and discharge (2.9±1.2,vs. 2.4±1.2, P=0.001). There were 4 in-hospital deaths (7.7%). The median follow-up was 21.0 months (ranging 1.5-111.0 months). Three patients (5.8%) were lost to follow-up, 1 patient required reoperation for mitral valve replacement and there was no death during follow-up. Significant improvement was seen in LVFS between discharge and final follow-up (32.1%±10.1% vs. 38.0%±6.0%, P=0.001); however, there was no significant difference in the degree of MR between discharge and final follow-up (2.4±1.2 vs. 2.3±1.2, P=0.541). There was no significant difference in cardiopulmonary bypass time, aortic cross-clamping time, mechanical ventilation time or hospital stay between the two groups. Conclusion Creation of a dual coronary system with reimplantation of the left coronary artery results in complete recovery of left ventricular function. However, concomitant management of MR during ALCAPA repair remains controversial. Concomitant mitral valve repair for ALCAPA patients with moderate-severe and severe MR is helpful to early function recovery of mitral valve.