In the study of repair of massive bone defect with free vascularized fibula graft, 13 cases were reported, in which traumatic defect in 7 cases, segmental resection of bone from tumors in 5 cases and osteomylitis in 1 cases. They all were treated successfully with vascularized fibular graft. After a follow-up of 6 months to 7 year, bone healing was observed with satisfactory and rehabilitation of functions. In one case, fatigued fracture occured twice due to early walking. It was concluded that free vascularized fibular graft was very helpful in the repair of massive bone defect, but prolonged external fixation after operation might be important to prevent fractur of grafted bone.
Objective
To investigate the effect of arginase (Arg) inhibitor N-ω-Hydroxy-L nor-Arginine (nor-NOHA) on high glucose cultured rhesus macaque retinal vascular endothelial cell line (RF/6A) in vitro.
Methods
The RF/6A cells were divided into the following 4 groups: normal control group (5.0 mmol/L of glucose, group A), high glucose group (25.0 mmol/L, group B), high glucose with 125 mg/L nor-NOHA group (group C), and high glucose with 1% DMSO group (group D). The proliferation, migration ability and angiogenic ability of RF/6A cells were measured by Methyl thiazolyl tetrazolium (MTT), transwell chamber and tube assay respectively. The express of Arg I, eNOS, iNOS mRNA of RF/6A cells were measured by real-time polymerase chain reaction (RT-PCR), Enzyme-linked immuno sorbent assay (ELISA) was used to detect the expression of NO and interleukine (IL)-1b of RF/6A cells.
Results
The proliferation, migration, and tube formation ability of group A (t=2.367, 5.633, 7.045;P＜0.05) and group C (t=5.260, 6.952, 8.875;P＜0.05) were significantly higher than group B. RT-PCR results showed the Arg I and iNOS expression in group B was higher than that in group A (t=6.836, 3.342;P＜0.05) and group C (t=4.904, 7.192;P＜0.05). The eNOS expression in group B was lower than that in group A and group C (t=4.165, 6.594;P＜0.05). ELISA results showed NO expression in group B was lower than that in group A and group C (t=4.925, 5.368;P＜0.05). IL-1b expression in group B was higher than that in group A and group C (t=5.032, 7.792;P＜0.05).
Conclusions
Nor-NOHA has a protective effect on cultured RF/6A cells in vitro and can enhance its proliferation, migration and tube formation. The mechanism may be inhibiting the oxidative stress by balancing the expression of Arg/NOS.
Abstract In order to have more selective sources of skin flaps to repair soft tissue defects, the prefabricated flap combining with skin expander was tried. Implanted the dorsal thoracic artery and vein with a muscle bundle of latissimus dorsi into the lateral thoracic wall subdermally andset a skin expander subcutaneously. Injected saline into the expander to inflate the flap gradually. In a month, an axial flap with the dorsal thoracic vesselswas prepared. the flap was transferred to the defect by vascular anastomosis technique. This method was applied in two cases, one to the left ankle, another to the left side of the neck. The sizes of the two flaps were 20cm×14cmand 22cm×15cm respectively. After operation, the flaps were alive completely. The advantages included selective source of vascular pedicle, thinpliable flap with enough blood supply, and direct closure of the donor site without skin graft.
ObjectiveTo explore repressive effects of transthyretitin (TTR) on the growth of human retinal endothelial cells (hREC) under high glucose and hypoxia environment.MethodshRECs were divided into 8 groups, including normal glucose group (5.5 mmol/L glucose), hypoxia group, high glucose group (25.0 mmol/L glucose), high glucose and hypoxia group, normal glucose group+TTR, normal glucose and hypoxia group+TTR, high glucose group+TTR, high glucose and hypoxia group+TTR. Flow cytometry was used to analyze cellular apoptosis. The expression level of Akt, p-Akt, eNOS, Bcl-2 and Bax protein were measured by Western blot.ResultsHypoxia could induce apoptosis as the apoptosis rate of normal and hypoxia group was higher than normal group (χ2=25.360, P＜0.05), high glucose and hypoxia group was higher that high glucose group (χ2=17.400, P＜0.05). The cell apoptosis rate of high glucose and hypoxia group+TTR were increased significantly as compared with high glucose and hypoxia group (χ2=9.900, P＜0.05). There was no statistically significant difference on the cell apoptosis rate between normal group and high glucose group, normal group+TTR and normal group, high glucose group+TTR and high glucose group, normal and hypoxia group+TTR and normal and hypoxia group (P＞0.05). Western blot showed that the expression of Akt did not change significantly in all eight groups（F=2.450, P＞0.05). Compared to normal group, the expression of p-Akt, eNOS, Bcl-2 in normal and hypoxia group were decreased (t=9.406, 5.306, 4.819), and the expression of Bax (t=?4.503) was increased (P＜0.05). Compared to high glucose group, same trend was found in high glucose and hypoxia group (t=8.877, 7.723, 6.500, ?14.646; P＜0.05). The expression of p-Akt in normal and hypoxia group+TTR was higher than normal and hypoxia group (t=?5.024, P＜0.05) , but there was no difference on the expression of eNOS, Bcl-2, Bax between these two groups (t=?2.235, ?2.656, ?0.272; P＞0.05). Compared to high glucose and hypoxia group, the expression of p-Akt and Bcl-2 in high glucose and hypoxia group+TTR were decreased (t=4.355, 4.308; P＜0.05), the expression of Bax was increased (t=?4.311, P＜0.05), and there was no difference on the expression of eNOS between these two groups (t=?1.590, P＞0.05). There was no statistically significant difference in the expression of p-Akt, eNOS, Bcl-2, Bax between high glucose group and normal group (t=?3.407, ?4.228, ?4.302, ?2.076; P＞0.05), normal group+TTR and normal group (t=?4.245, ?4.298, ?2.816, ?1.326; P＞0.05), high glucose group+TTR and high glucose group (t=4.016, ?0.784, 0.707, ?0.328; P＞0.05).ConclusionUnder high glucose and hypoxia, transthyretitin suppress the growth of hREC through Akt/Bcl-2/Bax, but not Akt/eNOS signaling pathway.
ObjectiveTo construct the connective tissue growth factor (CTGF) recombinant interference vector (shRNA) and observe its inhibitory effect on the expression of endogenous CTGF in retinal vascular endothelial cells. Methods The human CTGF shRNA was constructed and the high-titer CTGF shRNA lentivirus particles was acquired via three-plasmid lentivirus packaging system to infect retinal vascular endothelial cells. The optimal multiplicity and onset time of lentivirus infection were identified by tracing down the red florescent protein in interference vector. The cells were classified into three groups: blank control group, infection control group and CTGF knockdown group. The differences in cells migrating ability was observed through Transwell allay. The mRNA and protein expression of CTGF, fibronectin, α-smooth muscle actin (α-SMA) and collagen Ⅰ (Col Ⅰ) were quantified through real-time PCR testing and Western blot system. Data between the three groups were examined via one-way analysis of variance. ResultsThe result showed that an optimal multiplicity of 20 and onset time of 72 hours were the requirements to optimize lentivirus infection. Transwell allay result showed a contrast in the number of migrated cells in the CTGF knockdown group and that in the blank control group and infection control group (F=20.64, P=0.002). Real-time PCR testing showed a contrast in related gene expression (CTGF, fibronectin, α-SMA and Col Ⅰ) in the CTGF knocked-down group and that in the blank control group and infection control group (F=128.83, 124.44, 144.76, 1 374.44; P=0.000, 0.000, 0.000, 0.000). Western blot system showed the statistical significance of the contrasted number of related protein expression (CTGF, fibronectin, α-SMA and Col Ⅰ) in the knockdown group and that in the blank control group (F=22.55, 41.60, 25.73, 161.68; P=0.002, 0.000, 0.001, 0.000). ConclusionThe success in producing CTGF shRNA lentivirus particle suggests that CTGF shRNA lentivirus can effectively knock down CTGF expression.
Objective
To explore transthyretin (TTR) effect on retinal vascular endothelial cells (hREC) under high glucose and hypoxia environment.
Methods
hREC and human retinal pigment epithelial cell (hRPEC) were cultured at low-glucose (LG), high glucose (HG) and hypoxia. The glucose concentration was increased from 5.5 mmol/L up to 25 mmol/L, and hypoxia was induced by 200 μmol/L CoCl2. The cells were divided into LG group, LG-hypoxia group, HG group, HG-hypoxia group according to the different cell culture environment. The growth index was detected at 0, 4, 8, 16, 24, 36, 48, 60, 72 hours after cultured. Furthermore, hREC and hRPEC were also cultured with additional TTR (4 μmol/L), respectively. Then transwell co-culture system was employed to reveal the effects of hRPEC on the growth of hREC.
Results
At 72 hours after cultured, the growth index of hREC and hRPEC in LG group were increased as compared with LG-hypoxia group and HG group (hREC: F=17.098, 22.970; P < 0.05. hRPEC: F=45.442, 9.011; P < 0.05); the growth index of hREC and hRPEC were decreased in HG group and HG-hypoxia group (hREC: F=146.184, P < 0.05；hRPEC: F=27.907, P < 0.05). Additionally, hREC could be significantly repressed by added TTR during culture with high concentration of glucose (F=161.430, 24.106; P < 0.05). hREC could be significantly increased by added TTR during culture with low concentration of glucose (F=200.486, 48.662; P < 0.05). In co-culture process, hRPEC revealed inhibition activity against hREC under both natural and abnormal environment (LG group: F=15.711, P < 0.05; LG-hypoxia group: F=45.659, P < 0.05; HG group: F=7.857, P < 0.05; HG-hypoxia group: F=6.348, P < 0.05).
Conclusion
Under high glucose and hypoxia environment, the growth of hREC from neovascular could be inhibited by TTR.
Objective To study the clinicopathologic features which influence the prognosis of patients with stage Ib nonsmall cell lung cancer (NSCLC) after operation, and discuss the indication of postoperative chemotherapy. Methods From January 2002 to December 2002, the clinical materials of 152 patients who underwent complete pulmonary lobectomy and were confirmed to have stage Ib NSCLC by postoperative histopathological examination were collected from Shanghai Chest Hospital. There were 82 male and 70 female cases aged from 33-80 years. The mean age was 63.0 years. KaplanMeier method was used to compare and analyze the age, gender, tumor diameter, tumor location, lymphatic or vascular carcinoma embolus, differentiation, pleural invasion and chemotherapy of patients. Cox regression model was used to do prognostic multivariate analysis to above factors. Results The 5year survival rate was 71.1%. The median survival time was 44.20 months. The results of single factor analysis showed that the tumor diameter was longer than 5 cm（χ2=4.020，P=0.042）, lymphatic or vascular carcinoma embolus existed（χ2=14670，P=0.001）, poorly differentiated tumor（χ2=8.395，P=0.004）, and those whose tumors were located on middlelower lobars had a poor prognosis（χ2=3.980，P=0.045）. The age(χ2=0.478，P=0.740), gender(χ2=0.571，P=0.450), pathological type(χ2=0.406，P=0.816), pleural invasion(χ2=0.022，P=0.882) and postoperative chemotherapy of patients (χ2=1.067，P=0.302)had no relationship with postoperative survival. The results of multivariate analysis showed that lymphatic or vascular carcinoma embolus（P=0.006，95%CI:1.491,10.524） and poorly differentiated tumor（P= 0.001,95%CI:0.116,0.578） were the main factors which influenced the survival rate of patients. Conclusion The tumor differentiation and lymphatic or vessel carcinoma embolus of patients with stage Ib NSCLC are important factors which influence prognosis and survival rate. The poorly differentiated tumor and lymphatic or vessel carcinoma embolus could be regarded as one of the indications of postoperative chemotherapy.
Thumb and finger reconstruction by the method of pedal digit transplantation had been successfully performed in 541 casee from 1977 to 1996, which contained 404 cases of thumbs and 78 cases of fingers. The thumb reconstruction was mainly the simple transplantation of distal phalanx (42 cases) and the compound transplantation of hallucal nail-cutaneous flap with iliac bone segment (16 cases) for the defect of thumbs in degree 1 and 2. The combined transplantation of hallucal nail-cutaneous flap with the joint and tendons of the second toe (34 cases) and the transplantation of the distal part of the second toe (182 cases) for the defect of degree 3 and 4. The combined transplantation of the second pedal digit with its metatarsalphalangeal joint (189 caese) for the defect in degree 5 and 6. The finger reconstruction was performed by anastomosis of the arteries of the digit with those of the fingers for 29 cases with the defect in degree 2 and 3, 60 cases with the defect in degree 4 and 5, and 17 cases with the defect in degree 6. One-hundred and four cases of versels vasiation were found in this group (19 cases with the pedal dorsal artery, 13 cases with the greater saphenous vein and 72 cases with the first dorsal metatarsal artery). The main point of the operation and the treatment of the vessel variations were discussed.
OBJECTIVE: To sum up the experience of clinical application of distal base sural island flap. METHODS: From January 1997 to April 1999, the posterior island flap of leg pedicled with sural nerve and its nutrient vessels was applied to repair heel or dorsum of foot in 6 cases, chronic ulcer of heel in 2 cases, chronic osteomyelitis in 2 cases, scar contracture of bone defect accompanying fistulation in 1 case. The range of flap was 5 cm x 8 cm to 8 cm x 12 cm. RESULTS: All the flaps survived completely without vascular crisis. All the wounds healed by first intention. Followed up 3 to 12 months, no ulcer, osteomyelitis, fistulation were recurrence, and the sensation of flap was recovered slightly. CONCLUSION: The flap do not damage critical blood vessels and nerves, the donating region is hidden. The manual of operation is simple and blood supply of flap is sufficient. It can repair the defect of soft tissue on heel and dorsum with high survival rate.
Objective To investigate the change of vasa vasorum in vessel wall of varicose vein of the lower extre-mity. Methods Thirty-two patients with varicose vein of the lower extremity were collected， in which of 12 patients with simple varicose veins （varicose group）， 9 patients with recurrent varicose veins （recurrent group）， 11 patients withthrombophlebitis of varicose vein （thrombophlebitis group）， 9 patients with normal venous tissue as control group. HE staining was performed to observe the distribution of vasa vasorum and detect the vasa vasorum density. Results The increasing vasa vasorums were observed in the adventitia and media， but few was observed in the intima in the varicose， recurrent， and thrombophlebitis groups. The distribution of vasa vasorum was in the adventitia in the control group. The vasa vasorum densities （/mm2） in the varicose， recurrent， and thrombophlebitis groups （5.65±1.45，6.20±1.73， and 5.94±1.63， respectively） were greater than those in the control group （2.87±0.54）， the difference wasstatistically significant （P＜0.05）， but there was no significant difference of the vasa vasorum density among the varicosevein， recurrent， and thrombophlebitis groups （P＞0.05）. Conclusion Change of vasa vasorum is an important pathol-gical change with the nosogenis of varicose vein of the lower extremity.
