BACKGROUND: Studies have demonstrated that vascular endothelial cell growth factor (VEGF) and basic fibroblast growth factor (bFGF) play important roles in entochondrostosis and fracture healing blood vessel hyoperplasia. OBJECTIVE: To observe and compare the roles of VEGF and bFGF in fracture healing. DESIGN, TIME AND SETTING: Double-factor trial was performed at Pathological Laboratory, North China Coal Medical University from August 2005 to May 2006. MATERIALS: Twenty-four adult healthy Japan rabbits were selected and radial fracture models were created in 48 bilateral anterior limbs. The rabbits were randomly divided into VEGF group and bFGF group (n=24). METHODS: VEGF (0.2 ?g) and bFGF (100 ng) were injected respectively in the fracture site of rabbits. No external fixation was used. MAIN OUTCOME MEASURES: The rabbits were executed at postoperative weeks 2, 4 and 6 to measure callus sagittal diameter, transverse diameter and section area. X-ray was used to observe fracture healing and measure bony callus total area. Histological alterations during fracture healing were observed, and percentage of trabecular bone, cartilage and fibrous tissue in the bony callus was determined. RESULTS: At 2 weeks after operation, callus sagittal diameter and transverse diameter in VEGF group were significantly larger than bFGF group (P

BACKGROUND: The mouse model of cervical heart transplantation is an ideal medical research tool for study of transplant-induced ischemia/reperfusion injury and immunological rejection.However,technical problems have limited the widespread use of mouse cervical vascularized heart transplantation.OBJECTIVE: To improve the cervical heterotopic heart transplantation in mice using the tail-cuff technique.METHODS: Isogeneic transplantation was performed from Balb/c to BALB/c mice,and allogeneic transplantation from C57BL/6 to BALB/c mice.The right common carotid artery and the external jugular vein of the recipient were equipped with a tail cuff made from 24 G and 22 G intravenous catheter,and everted over the cuff,and then connected with the aorta and the pulonary artery of donor heart,respectively.RESULTS AND CONCLUSION: A total of 36 transplants for formal experiment,12 for isogeneic transplantation,and 24 for allogeneic transplantation,were performed with a surgical successful rate of 100%.The total surgical procedure was(49.6±7.4)minutes and total ischemic time of the grafts was(28.8±4.2)minutes.In particular,the average time for vascular everting and for the reconnection of both vessels was obviously shortened.This improved tail-cuff technique shows its superiority,and can serve as an ideal method for establishing cervical heterotopic heart transplantation model in mice.

0.05).The blood gas analysis(BGA) and pulmonary function test(PFT) were performed in patients with stable COPD.The cognitive function was evaluated by ESD in both patients with stable COPD and healthy persons as control.The patients were divided into different groups by BGA and PFT(FEV1/Pred).Results:The total score of ESD and the subtest scores of ESD in leaning,memory,calculation,constructive function were obviously lower in the patient group than those in the control group(The mean difference of the total score was 16,with the two groups' total scores as 208.1?17.6/224.3?10.6,t=5.19,P

Objective To investigate a new method for the reconstruction of the urethra using peritoneal free grafts in phalloplasty in a rabbit model. Methods Animal models were established in 24 adult male New Zealand white rabbits, which were randomly divided into the peritoneal graft group (n=12) and peritoneum-skin graft group (n=12). In the former group the peritoneal tube grafts with the mesothelial surface inward were used as the urethral substitutes. And a circumferential 1 cm rim of scrotal skin was inserted at the tip of the tubularized peritoneal graft for urethroplasty in the latter group. Subsequently, a superficial epigastric fasciovascular pedicle flap for phalloplasty was harvested and tubed over the reconstructed urethra. The process of growth was observed grossly. Results The reconstructed penis survived well without urethral stricture, and the peritoneal-lined grafts survived in all rabbits with a smooth, moist quality and without ulceration and fibrosis. Gross examination showed 8 meatal occlusions with fistulas simultaneously in the peritoneal graft group, and 3 fistulas in the peritoneum-skin graft group. Conclusion The authors have successfully designed the rabbit model of the urethroplasty using the tubularized peritoneal free grafts in the penile reconstruction and demonstrated the possibility of the new method for phalloplasty brings a light to clinical study.

AIM: To detect dystrophin expression in skeletal muscles of mdx mice after bone marrow transplantation (BMT), and to evaluate the effect of BMT on Duchenne muscular dystrophy (DMD). METHODS: Bone marrow cells were cultured for three days, and then transplanted into mdx mice irradiated lethally through tail veins. After 4 and 6 months, dystrophin expression on myocytes membranes in mdx mice was detected by fluorescent immunohistochemical staining. The centrally nucleated fibers (CNF) were calculated by HE staining, and the physiologic parameters measured and the motor function detected by traction test, rotating rods test and rotating wheels test were also observed. RESULTS: Until 4 and 6 months after BMT, dystrophin was expressed partly on myocytes membranes in mdx mice, and the ratio of CNF decreased, physiologic functions improved, the motor ability reinforced in treated group. CONCLUSION: After BMT, marrow stem cells settled in injured skeletal muscles and bone marrow, then differentiated into myocytes with dystrophin expression and caused the improvement of pathology, physiology and motor function in treated group finally. These results give a powerful proof for the treatment of DMD with BMT.

AIM: To study the effect of bone marrow stem cell transplantation on mdx mice at different ages. METHODS: The bone marrow stem cells of C57BL/6 mice (4 - to-weeks age) were cultured in vitro for 3 days, then injected intravenously into the 6 -week and 8-week aged mdx, which were preconditioned with 7 Gy ? ray. 12 weeks after being transplanted, the mdx mice were studied for the dystrophin protein expression on the skeletal muscle membrane. RESULTS: Three months after transplanted with bone marrow stem cells, about 16% and 7% muscles cells in 6-week and 8-week mdx mice expressed dystrophin protein, respectively. CONCLUSION: 12 weeks after transplantation with bone marrow stem cells of homologous series mice, different amounts of dystrophin protein expressed on the membrane of skeletal muscle cells were observed in different aged mdx mice. Bone marrow stem cell transplantation show more benefic effect for younger mdx mice.

AIM: To observe the effects of overload exercise on skeletal muscles in X-linked muscular dystrophy(mdx) mice.METHODS: Mdx mice and C57 mice were carried out swimming and hanging tail movement tests (mdx mice as control did not exercise). It lasted for 13 minutes each time per day, and lasted 3 days. Evans blue was injected into tail vain. The mice were killed the next day, and the hind limbs were taken photographs after skins were flayed. The gastrocnemius muscles and diaphragms cryostat sections were made. Under a fluorescence microscope, Evans blue staining was seen. Then the sections were tested by routine HE staining, the histological change of muscles was analyzed under a light microscope.RESULTS: Many blue colored longitudinal lines were observed in skeletal muscles of mdx mice, whereas they were hardly seen in control mdx and C57 mice. Under a fluorescence microscope, some muscle fibers of mdx mice were stained with Evans blue, few muscle fibers of control mdx mice were stained, and C57 mice were not. Under a light microscope, HE staining of muscles showed some degenerated muscle fibers became round in shape and the myonuclei became condensed, or necrotic fibers had amorphous structures, most of them in the degenerated and necrotic fibers of diaphragms C57 mice did not have these changes.CONCLUSION: Overload exercise did harm to skeletal muscles of mdx mice; Vital staining with Evans blue is useful not only for distinguishing degenerating muscle fibers, but also for studying the degeneration process in dystrophin-deficient muscle.

Objective To investigate the effect of chloroform extract of Zingiber mioga Roscoe on human dermal microvascular endothelial cell (HDMEC) expression of cell adhesion molecules and adhesivity to lymphocytes.Methods HDMECs were isolated from human foreskin tissue and subjected to subculture.After several passages,some HDMECs were treated with chloroform extract from Zingiber mioga Roscoe (CFMG) of 200 mg/L for 30 minutes before or after 24-hour stimulation with tumor necrosis factor (TNF)-α or interleukin (IL)-1α.Subsequently,enzyme-linked immunosorbent assay was performed to determine the expression levels of intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and E-selectin on HDMECs.Both T lymphocytes isolated from venous blood of healthy adults and Ramos Burkitt's lymphoma B cells were labelled with chromium-51 and incubated with the HDMECs for four hours followed by the determination of radiation intensity of lymphocytes adhering to HDMECs using a gamma-counter.Results Compared with 0.2％ dimethyl sulfoxide (DMSO),CFMG slightly down-regulated the expression of ICAM-1,VCAM-1 and E-selectin on resting HDMECs (all P ＞ 0.05).In comparison with culture medium,TNF-α enhanced the expressions of ICAM-1,VCAM-1 and E-selectin significantly (all P ＜ 0.01),and IL-1α elevated the expressions of ICAM-1 and E-selectin markedly(both P ＜ 0.01) as well as the expression of VCAM-1 slightly (P ＞ 0.05).The upregulating effects of TNF-α and IL-1α on the expressions of adhesion molecules were notably suppressed by the CFMG treatment prior to the stimulation with TNF-α and IL-1α(all P ＜ 0.01),but not affected by that after the stimulation (all P ＞ 0.05).The adhesivity of HDMECs to T lymphocytes and Ramos cells was slightly decreased by CFMG treatment compared with 0.2％ DMSO (both P ＞ 0.05),but was markedly increased by TNF-α and IL-1α compared with the culture medium (both P ＜ 0.01).Conclusions CFMG may play an antiinflammatory role via blocking the up-regulating effect of pre-inflammatory cytokines such as TNF-α and IL-1α on the expression of adherence molecules on HDMECs and,hence,inhibiting inflammatory cell infiltration in tissue.

Objective To investigate the role of induction of carbon monoxide (CO) with methylene chloride (MC) in recipients in ameliorating allograft rejection and prolonging allograft survival and to explore the possible mechanisms in a murine heart transplantation model.Methods Inbred male C57BL/6 mice were used as donors and inbred male Balb/c mice as recipients respectively to establish cervical heterotopic heart transplantation model.The experiments were divided into 3 groups.Recipients were treated with MC (100 mg/kg,per os) day 1 prior to transplantation to day 6 posttransplantation (group MC1w) or to day 13 posttransplantation (group MC2w),or treated with isovolumic olive oil day 1 prior to transplantation to cardiac arrest of allograft (group Tx).The serum TNF-α and IL-10 proteins,TNF-α and IL-10 mRNA,and Foxp3 mRNA and protein in cardiac grafts were measured respectively.The intercellular adhesion molecule-1 (ICAM-1) and Caspase-3 protein in cardiac grafts,and the histopathologic changes of cardiac grafts were also observed.Results Serum COHb levels in untreated mice were (0.85 ± 0.28)％.After MC application,serum COHb peaked within 3 h in recipients (5.24 ± 0,45)％ (P＜0.01 ).The median survival time of cardiac grafts in group MC1w(12.1 days) and group MC2w( 19.4 days) was longer than that in group Tx (6.3 days) (P ＜0.01). As compared with group Tx,induction of CO in group MC1w and group MC2w down-regulated significantly the levels of serum TNF-α (P＜0.01 ) and TNF-α mRNA (P＜0.01) of cardiac grafts and spleen in recipient mice,inhibited the protein expression of ICAM-1 (P＜0.01) and Caspase-3 (P＜0.01) of cardiac grafts,and inhibited,especially in group MC2w,the proliferation of lymphocytes and monocytes infiltration in cardiac grafts.There was no significant difference in serum IL-10 and Foxp3 mRNA and protein in cardiac grafts and spleen of recipients among the groups (P＞0.05).Conclusion Induction of CM in recipients could relieve cardiac allograft rejection and prolong cardiac allograft survival via its anti-inflammation and anti-apoptotic effects,not via up-regulation of Foxp3 in recipient mice.