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Objective To explore the relationship between latissimus dorsi myocutaneous flap and blood supply, so as to provide a scientific basis for the re-division, transposition and transplantation of latissimus dorsi myocutaneous flap. Methods The latissimus dorsi muscle of 48 cadavers were observed by anatomy and angiography. The clinical applications of latissimus dorsi myocutaneous flap in 31 cases were reviewed. Results The latissimus dorsi myocutaneous flap had many sources of blood supply. The main thoracodorsal artery was distributed in the upper and outer latissimus dorsi muscle. The medial and lateral branches were separated steadily with their respective distribution areas. The inner and lower parts of latissimus dorsi muscle were supplied by intercostal and lumbar arteries. The anastomosis between them and the branches of thoracodorsal artery was obvious according to X-ray angiography. The caliber was between 320-550 μm. The blood supply of the skin superficial to the latissimus dorsi muscle was from the myocutaneous artery. But the anastomosis between the perforating branches was sparse and the caliber was small near the inner and lower parts. Myocutaneous flaps were applied for wound repair, breast reconstruction and leg defect repair after mass excision in 31 cases. Thirty cases of myocutaneous flaps survived completely post-operation. One case of myocutaneous flap had early signs of partial blood flow disturbance. After treatment, blood flow gradually improved and wound healing delayed. Conclusion The latissimus dorsi myocutaneous flap can be divided into 3 parts according to their arterial branches and anastomotic characteristics, which can provide the vascular anatomical basis for whole or partial separation, transposition or transplantation and preservation of muscle function.
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OBJECTIVE@#To investigate the outcomes of breast reconstruction with employing improved techniques throughout the tissue expander/implant two-stage breast reconstructed process, which involved the tissue expander placement, the saline filling intraoperatively and postoperatively, the implant selection, and the permanent implant replacement.@*METHODS@#In this study, 68 patients who had been provided immediate or delayed tissue expander/implant two-stage breast reconstruction with autologous fat injection post-mastectomy in Peking University Third Hospital from April 2014 to September 2018 were involved, and the relevant information was analyzed retrospectively. The enhancements of the techniques, involving the incision selection, the expander placement, the principle of expansion, the management of capsule, the prosthesis selection, and the assisted reconstruction method were summarized, and the reconstruction outcomes were evaluated objectively through three-dimensional surface imaging.@*RESULTS@#Among the 68 patients in this study, immediate reconstruction was conducted in 25 patients and 43 patients underwent delayed reconstruction. The median time of tissue expansion was 7.0 (3.0, 20.0) months, and the average volume of expansion was (372.8±87.2) mL. The median size of breast implant was 215 (100, 395) mL. The median number of injections for fat grafting was 1 (1, 3), and the average volume of fat grafting was (119.3±34.1) mL. The median follow-up time was 7.0 (4.0, 24.0) months. During the process of breast reconstruction, the tissue expander leakage was observed in two patients, and one of them underwent expander replacement due to the secondary infection. In the immediate reconstruction cases, the volume symmetry of bilateral breasts after reconstruction got even better than that before mastectomy (t=4.465, P<0.01). And in the delayed reconstruction cases, the volume between bilateral breasts also achieved good symmetry after reconstruction (t=0.867, P>0.1).@*CONCLUSION@#Good results of tissue expander/implant two-stage breast reconstruction could be achieved through the techniques enhancement, which involved the preferred transverse incision, the downward placement of expander, the rapid expansion of chest soft tissue, the release of capsule tension, the application of sizer in prosthesis selection, and the assisted autologous fat grafting.
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Humanos , Neoplasias da Mama , Mamoplastia , Mastectomia , Estudos Retrospectivos , Dispositivos para Expansão de Tecidos , Resultado do TratamentoRESUMO
The activation of hepatic stellate cells (HSCs) and their transformation to myofibroblasts are the key steps in the pathological progress of liver fibrosis. The transforming growth factor-β (TGFβ)/Smad pathway is involved in the proliferation and collagen synthesis of HSCs. This study aimed to examine the effect of the protease inhibitor MG132 on the signaling pathway of TGFβ/Smad in HSC-T6 cells and seek a novel therapeutic approach for liver fibrosis. The HSC-T6 cells were treated with MG132 at different concentrations (0-10 μmol/L). Cell proliferation was detected by MTT method. The mRNA and protein expression levels of TGFβ1, Smad3 and Smad7 were determined in HSC-T6 cells by real-time PCR and Western blotting, respectively, after treatment with MG132 at different concentrations (1, 2, 3 μmol/L) or RPMI1640 alone (serving as control). The results showed that MG132 could inhibit the proliferation of HSC-T6 cells in a dose-dependent manner, and the IC(50) of MG132 was 6.84 μmol/L. After treatment with MG132 at 1, 2 or 3 μmol/L for 24 h, the mRNA expression levels of TGF-β1 and Smad3 were significantly decreased (P<0.05), but the Smad7 mRNA expression had no significant change (P>0.05). There was also a significant decrease in the protein expression level of TGF-β1 and Smad3 (P<0.05). However, the expression of Smad7 protein was substantially increased when compared with the control group (P<0.05). It was concluded that the inhibition of TGFβ/Smad pathway in HSC-T6 cells by MG132 can reduce the production of profibrosis factors (TGFβ1, Smad3) and promote the expression of anti-fibrosis factor (Smad7), suggesting that MG132 may become a potential therapeutic alternative for liver fibrosis.
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Animais , Ratos , Linhagem Celular , Leupeptinas , Farmacologia , Inibidores de Proteases , Farmacologia , Transdução de Sinais , Proteínas Smad , Metabolismo , Fator de Crescimento Transformador beta , MetabolismoRESUMO
The activation of hepatic stellate cells (HSCs) and their transformation to myofibroblasts are the key steps in the pathological progress of liver fibrosis. The transforming growth factor-β (TGFβ)/Smad pathway is involved in the proliferation and collagen synthesis of HSCs. This study aimed to examine the effect of the protease inhibitor MG132 on the signaling pathway of TGFβ/Smad in HSC-T6 cells and seek a novel therapeutic approach for liver fibrosis. The HSC-T6 cells were treated with MG132 at different concentrations (0-10 μmol/L). Cell proliferation was detected by MTT method. The mRNA and protein expression levels of TGFβ1, Smad3 and Smad7 were determined in HSC-T6 cells by real-time PCR and Western blotting, respectively, after treatment with MG132 at different concentrations (1, 2, 3 μmol/L) or RPMI1640 alone (serving as control). The results showed that MG132 could inhibit the proliferation of HSC-T6 cells in a dose-dependent manner, and the IC50 of MG132 was 6.84 μmol/L. After treatment with MG132 at 1, 2 or 3 μmol/L for 24 h, the mRNA expression levels of TGF-β1 and Smad3 were significantly decreased (P0.05). There was also a significant decrease in the protein expression level of TGF-β1 and Smad3 (P<0.05). However, the expression of Smad7 protein was substantially increased when compared with the control group (P<0.05). It was concluded that the inhibition of TGFβ/Smad pathway in HSC-T6 cells by MG132 can reduce the production of profibrosis factors (TGFβ1, Smad3) and promote the expression of anti-fibrosis factor (Smad7), suggesting that MG132 may become a potential therapeutic alternative for liver fibrosis.
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The type I interferon and IFNAR play an important role in hepatitis B virus (HBV) infection and anti-HBV therapy. However, its mechanism of action is still poorly understood. To gain more insights into the role of type I interferon and type I interferon receptor (IFNAR) in HBV infection, we established an HBV persistent replication IFNAR knockout (IFNAR(-/-)) mouse model and preliminarily applied this model. At first, the progeny of IFNAR(-/-) mouse was reproduced. Then hydrodynamic injection with pAAV/HBV1.2 plasmid was conducted to establish the persistent HBV replication IFNAR(-/-) mouse model. At last, we applied this model to evaluate the effect of nucleoside analogues entecavir (ETV) on HBV replication. It was found that there was no difference in the serum HBsAg and HBeAg levels and HBcAg expression in the liver tissue between the ETV treated groups and normal saline (NS) treated group, but the serum HBV DNA levels were significantly suppressed 10, 25, 40 and 55 days after the ETV treatment [P=0.035, P=0.00, P=0.149 and P=0.084, IFNAR knockout (KO) control group vs. C57BL/6 ETV groups, respectively; P=0.081, P=0.001, P=0.243 and P=0.147, IFNAR KO control group vs. IFNAR KO ETV groups, respectively]. Interestingly, there was no difference in serum HBV DNA levels between the ETV treated IFNAR(-/-) and C57BL/6 mice. This result suggests that HBV suppression during ETV treatments doesn't depend on type I interferon and IFNAR. Collectively, persistent HBV replication IFNAR(-/-) mouse model that we established is a useful and convenient tool to detect the function of the type I interferon and IFNAR in HBV infection and anti-HBV treatments.
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The type I interferon and IFNAR play an important role in hepatitis B virus (HBV) infection and anti-HBV therapy. However, its mechanism of action is still poorly understood. To gain more insights into the role of type I interferon and type I interferon receptor (IFNAR) in HBV infection, we established an HBV persistent replication IFNAR knockout (IFNAR(-/-)) mouse model and preliminarily applied this model. At first, the progeny of IFNAR(-/-) mouse was reproduced. Then hydrodynamic injection with pAAV/HBV1.2 plasmid was conducted to establish the persistent HBV replication IFNAR(-/-) mouse model. At last, we applied this model to evaluate the effect of nucleoside analogues entecavir (ETV) on HBV replication. It was found that there was no difference in the serum HBsAg and HBeAg levels and HBcAg expression in the liver tissue between the ETV treated groups and normal saline (NS) treated group, but the serum HBV DNA levels were significantly suppressed 10, 25, 40 and 55 days after the ETV treatment [P=0.035, P=0.00, P=0.149 and P=0.084, IFNAR knockout (KO) control group vs. C57BL/6 ETV groups, respectively; P=0.081, P=0.001, P=0.243 and P=0.147, IFNAR KO control group vs. IFNAR KO ETV groups, respectively]. Interestingly, there was no difference in serum HBV DNA levels between the ETV treated IFNAR(-/-) and C57BL/6 mice. This result suggests that HBV suppression during ETV treatments doesn't depend on type I interferon and IFNAR. Collectively, persistent HBV replication IFNAR(-/-) mouse model that we established is a useful and convenient tool to detect the function of the type I interferon and IFNAR in HBV infection and anti-HBV treatments.
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Animais , Feminino , Humanos , Masculino , Camundongos , Doença Crônica , Modelos Animais de Doenças , Hepatite B , Genética , Virologia , Vírus da Hepatite B , Fisiologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptor de Interferon alfa e beta , Genética , Metabolismo , Replicação Viral , GenéticaRESUMO
<p><b>OBJECTIVE</b>This report aims to investigate the Toll-like receptor (TLR) signaling pathways and induced antiviral activity in hepatocytes.</p><p><b>METHODS</b>We isolated primary hepatocytes from wild-type C57BL/6 mice and examined the expression of TLR by realtime RT-PCR. Hepatocytes were stimulated with TLR 1-9 agonists and the supernatants were harvested. The secretion of cytokines were tested by ELISA. The antiviral effectors in supernatants were assayed via virus protection assay (in EMCV system) and the control of HBV replication were assessed via Southern blotting (in HBV system).</p><p><b>RESULTS</b>We demonstrated that hepatocytes expressed TLR1-9. In accordance with these TLR expression profiles, hepatocytes responded to all TLR ligands by producing inflammatory cytokines (TNF-α or IL-6), to TLR -1,-3,-7 and -9 ligands by producing type I IFN (IFN-α or IFN-β). Only TLR 3 and TLR 7 agonists could stimulate the production of high amounts of antiviral mediators by hepatocytes in virus protection assay. By contrast, supernatants from TLR1, -3 and -4 directly stimulated hepatocytes and TLR 3, -7 and -9 transfected hepatocytes were able to potently suppress HBV replication.</p><p><b>CONCLUSION</b>Primary hepatocytes display a unique TLR signaling pathway and can control HBV replication after stimulation by TLR agonists in mice. It may be helpful for the development of TLR-based therapeutic approaches against hepatotropic virus.</p>
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Animais , Camundongos , Células Cultivadas , Vírus da Hepatite B , Alergia e Imunologia , Fisiologia , Hepatócitos , Alergia e Imunologia , Metabolismo , Imunidade Inata , Camundongos Endogâmicos C57BL , Transdução de Sinais , Receptores Toll-Like , Alergia e Imunologia , Metabolismo , Replicação ViralRESUMO
<p><b>OBJECTIVE</b>To illustrate the effects of the site and method of liposuction on differentiation of human preadipocytes.</p><p><b>METHODS</b>Forty-two fatty samples were obtained with liposuction, which were then divided into four groups according to operation sites (abdomen, hip or extremity) and the methods (conventional negative-pressure or syringe method). Each sample was treated with collagenase I to release preadipocytes for in vitro culture. Affected by the differentiation-induced agents for 9 days or 28 days, the cultured adipocytes were stained with Nile red and observed under a fluorescent microscope. The differentiation rates were examined with flow cytometric analysis and the quantity of intracytoplasmic lipids was determined with oil red O staining. The results were analyzed with independent samples t-test (Mann-Whitney) using SPSS 10.0.</p><p><b>RESULTS</b>There was no statistical difference in differentiation at the 9th day or 28th day among the preadipocytes obtained from the abdomen, hip or extremity with the negative-pressure method. The preadipocytes from the abdomen differentiated more at the 28th day than the 9th day (P < 0.05), which was not observed in the hip or the extremity. The preadipocytes obtained from the abdomen with the negative-pressure method differentiated more than those with the syringe method (P < 0.05).</p><p><b>CONCLUSION</b>No essential difference was found in preadipocyte differentiation among the liposuction sites, while the abdomen might have some superiority. The negative-pressure method of liposuction is the first option in future research of tissue engineering. The flow cytometric analysis is a convenient way to study preadipocyte differentiation.</p>