Y-box-binding protein 1 mediates sorafenib resistance via the extracellular signal regulated-protein kinase pathway in hepatoma cells / 中华肝脏病杂志

Objective: To investigate the effect and possible mechanism of Y-box-binding protein 1 (YB-1) on sorafenib resistance in hepatoma cells. Methods: Lentiviral vectors with YB-1 overexpression and knockdown were constructed, respectively, to stimulate human hepatoma cell lines (HepG2 and Huh7) alone or in combination with sorafenib.The overexpression part of the experiment was divided into four groups: overexpression control group (Lv-NC), YB-1 overexpression group (Lv-YB-1), overexpression control combined with sorafenib resistance group (Lv-NC+sorafenib), YB-1 overexpression combined with sorafenib resistance group (Lv-YB-1 + sorafenib). The knockdown part of the experiment was also divided into four groups: knockdown control group (Lv-shNC), YB-1 knockdown group (Lv-shYB-1), knockdown control combined with sorafenib resistance group (Lv-shNC + sorafenib), YB-1 knockdown combined with sorafenib resistance group (Lv-shYB-1 + sorafenib). The occurrence of cell apoptosis was detected by TUNEL. The protein expression levels of phosphorylated (p)-ERK and ERK, key proteins in the extracellular regulatory protein kinase (ERK) signaling pathway, were detected by Western blot and quantified by ImageJ software. Subcutaneous tumorigenesis experiments were performed in nude mice. The effect of YB-1 on the efficacy of sorafenib was verified in vivo. The comparison between the two sets of data was carried out by an independent sample t-test. One-way ANOVA was used for comparisons between the three groups of data above. Results: Sorafenib had accelerated the occurrence of apoptosis in hepatoma cells, while YB-1 overexpression had inhibited cell apoptosis, and at the same time also inhibited the apoptosis-accelerating impact of sorafenib. On the contrary, YB-1 knockdown accelerated cell apoptosis and amplified the induction effect of sorafenib on apoptosis. Furthermore, sorafenib resistance had down-regulated p-ERK levels (HepG2: Lv-NC 0.685 ± 0.143, Lv-NC + sorafenib 0.315 ± 0.168, P < 0.05; Huh7: Lv-NC 0.576 ± 0.078, Lv-NC + sorafenib 0.150 ± 0.131, P < 0.01), whereas YB-1 overexpression had inhibited sorafenib resistance p-ERK reduction (HepG2: Lv-NC + sorafenib 0.315 ± 0.168, Lv-YB-1 + sorafenib 0.688 ± 0.042, P < 0.05; Huh7: Lv-NC + sorafenib 0.150 ± 0.131, Lv-YB-1 + sorafenib 0.553 ± 0.041, P < 0.05). YB-1 knockdown further increased sorafenib-induced p-ERK downregulation (HepG2: Lv-shNC + sorafenib 0.911 ± 0.252, Lv-shYB-1 + sorafenib 0.500 ± 0.201, P < 0.05; Huh7: Lv-shNC + sorafenib 0.577 ± 0.082, Lv-shYB-1 + sorafenib 0.350 ± 0.143, P < 0.05), which was further verified in naked mice (Lv-shNC + sorafenib 0.812 ± 0.279, Lv-shYB-1 + sorafenib 0.352 ± 0.109, P < 0.05). Conclusion: YB-1 mediates the occurrence of sorafenib resistance via the ERK signaling pathway in hepatoma cells.

The mechanisms of inhibitory effect of adenovirus-mediated wild-type PTEN gene on the proliferation in activated hepatic stellate cells in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Using an adenoviral vector, the wild-type PTEN gene was transduced into activated hepatic stellate cell (HSC) cultured in vitro and cell cycle markers and were detect. Thereby, the potential mechanisms of inhibitory effect of the wild-type PTEN overexpression on the proliferation in activated HSC was investigated.</p><p><b>METHODS</b>The wild type PTEN gene was transduced into activated HSC (HSC-T6 ) cultured in vitro mediated by adenoviral vector. PTEN expression in HSC was measured by Western blot and Real-time fluorescent quantitation PCR. Flow cytometry (FCM) was then used to detect cell cycle phase of activated HSC. And the expressions of cyclinD1 and cyclin dependent kinase 4 (CDK4) in HSC were determined by Western blot.</p><p><b>RESULTS</b>The data showed that exogenous wild type PTEN gene was successfully transduced and expressed in activated HSC cultured in vitro. The over-expression of wild type PTEN resulted in the increased number of HSC at G0/G1 phase ( P less than 0.01), and the number of HSC at S phase and G2/M phase were decreased significantly, P less than 0.01. Furthermore, there were decreased cyclinD1 and CDK4 expression in HSC infected with Ad-PTEN, P less than 0.01.</p><p><b>CONCLUSION</b>The over-expression of wild type PTEN inhibit transition of activated HSC in vitro from G1 to S phase and arrest cell cycle of them at G0/G1 phase via the down-regulated expressions of cyclinD1 and CDK4, and then inhibit HSC proliferation.</p>

Treatment of complicated cavernous venous malformation with retained copper wires combined with pingyangmycin injection / 中华整形外科杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of retained copper wires combined with pingyangmycin (PYM) injection for complicated cavernous venous malformation.</p><p><b>METHODS</b>The location of venous malformation was detected by physical examination and MRI. The copper wires in 0.2 mm width were used to puncture the lesion repeatedly and retained in the lesion to form a net. After that, 8 mg PYM was injected into the residue malformed veins. 8-10 days later, the copper wires were taken out and necrotic tissue was squeezed out. The wounds of punctual holes healed through dressing. The patients received postoperative MRI to evaluate the therapeutic effect.</p><p><b>RESULTS</b>From Jan. 2002 to Dec. 2008, 45 cases were treated. The patients were followed up for 1-3 years. 51.1% (23/45) of the lesions shrinked markedly or even disappeared. 42.2% (19/45) of the lesions reduced. 6.67% (3/45) of the lesions didn't change. There was no complication like invasive infection.</p><p><b>CONCLUSIONS</b>It is very effective to treat complicated cavernous venous malformation with retained copper wires combined with pingyangmycin injection.</p>

Central wedge resection and asymmetric Z-plasty for minor labia reduction / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effect of central wedge resection and asymmetric Z-plasty for minor labia reduction.</p><p><b>METHODS</b>Based on the Giraldo procedure, the incision was designed and the redudent tissue was resected quantitatively. The Z-plasty was modified to rectangle flap with deviated incision. The incisions at the two surface of minor labia were designed in an opposited direction. The two rectangle flaps were inserted to form the free edge of minor labia.</p><p><b>RESULTS</b>11 cases of minor labia hypertrophy were treated with good results.</p><p><b>CONCLUSIONS</b>The modified procedure is easily performed with precise design. It is suitable for all kinds of minor labia hypertrophy.</p>

The expression and role of glucose transporter-1 in infantile hemangioma / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and role of glucose transporter-1 (Glut1) in infantile hemangioma.</p><p><b>METHODS</b>Fifty-two samples from infantile hemangioma, 25 in cavernous venous malformation, 9 in arteriovenous malformation, 2 in capillary malformation and 5 in normal skin samples were involved in this study. The EnVision immunohistochemical stain was used to investigate the expression of Glut1 protein in these samples.</p><p><b>RESULTS</b>In the early proliferating stage, a number of endothelial cells expressed Glut1. In the middle proliferating stage, most of vascular endothelial cells and scattered endothelial cells expressed Glut1. In the late proliferating stage, the expression of Glut1 decreased quickly. In the involuting stage, all hemangioma samples didn't express Glut1. All of the samples from the cavernous venous malformations, arteriovenous malformations, capillary malformations and normal skin had no expression of Glut1.</p><p><b>CONCLUSIONS</b>Glut1 may be one of the phenotypes of infantile hemangioma endothelial cells in their development, rather than the inherent character. The expression of Glut1 changes according to the metabolic need of infantile hemangioma cells.</p>

Plastic treatment of Paget's disease of the scrotum and penis / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the clinical features and plastic treatment of Paget's disease of the scrotum and penis.</p><p><b>METHODS</b>We analyzed 11 cases of Paget's disease of the scrotum and penis treated from 1997 to 2007. Extended excision of the focus of infection was performed and the skin absence was repaired by free skin grafting, local random skin flap, and island skin flap.</p><p><b>RESULTS</b>Satisfactory wound healing was achieved in all but 1 case, which was delayed due to infection. All the patients were followed up for 1-4 years. One patient relapsed 2 years after the surgery and received a second operation, and another 1 died of lymphoma complicated by lung infection. The original shape and contractibility of the scrotum and penis were basically preserved and their appearance and function were fairly good.</p><p><b>CONCLUSION</b>For patients with Pagets disease of the scrotum and penis, it is a desirable method to at once excise the lesion and repair the skin absence by plastic surgery.</p>

Effect of Salvia miltiorrhiza monomer IH764-3 on MMP-13 and TIMP-1 by downregulating the expression of focal adhesion kinase in hepatic stellate cell stimulated by H2O2 / 中国应用生理学杂志

<p><b>AIM</b>To investigate the effect of IH764-3 on the expression of MMP-13 and TIMP-1 by H2O2-stimulated hepatic stellate cell and the alteration of FAK during this process.</p><p><b>METHODS</b>The expression of MMP-13 and FAK mRNA was examined by RT-PCR. TIMP-1 mRNA was analyzed by in-situ hybridization. FAK and TIMP-1 were evaluated at protein level through Western blotting method.</p><p><b>RESULTS</b>Being incubated for 2 h, compared with control group, MMP-13 mRNA was upregulated by IH764-3, but TIMP-1 transcription was reduced in a dose-dependent manner, accompanied with the decrease of FAK mRNA. The expression of TIMP-1 and FAK protein in HSC also decreased after being exposed by IH764-3 for 24 h.</p><p><b>CONCLUSION</b>IH764-3 can induce the expression of MMP-13 and inhibit the expression of TIMP-1. Down-regulating the expression of FAK mRNA may be one of its mechanisms.</p>

The distribution and evolution of pericytes in infantile hemangioma / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the distribution, phenotype and development of pericytes in infantile hemangioma.</p><p><b>METHODS</b>Fifty-two infantile hemangioma samples were included in our study. alpha-SMA was used as the marker antigen to observe the distribution of pericytes. Transmission electron microscope and TUNEL method were used to analyze the apoptosis of pericytes.</p><p><b>RESULTS</b>In the early and middle proliferating stage, there existed many pericytes in hemangioma; Pericytes together with endothelial cells generated vasculogenesis. In the late proliferating stage, many pericytes became apoptotic. In the early involuting stage, there were only a few of pericytes around the microvessels; After that, the microvessels became obstruction progressively and pericytes disappeared finally.</p><p><b>CONCLUSIONS</b>The pericyte is one of the major constitutive cells of hemangioma. The vasculogenesis, development and disappearance of microvessels undertaken by pericytes and endothelial cells lead to the pathologic evolution of infantile hemangioma.</p>

Fasudil inhibits HSC adhesion, migration and proliferation via Rho/ROCK pathway / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To observe the effects of fasudil, a Rho/ROCK signaling pathways inhibitor, on adhesion, migration and proliferation of hepatic stellate cells (HSCs).</p><p><b>METHODS</b>Cultured HSCs were divided into 5 groups. Fasudil was added to 4 groups in a concentration of 12.5, 25, 50, and 100 micromol/L, respectively. A group without fasudil added served as untreated controls. The adhesive inhibition effect of fasudil on HSCs was examined by toluidine blue colorimetric assay, the inhibition of migration of HSCs was evaluated by a modified Boyden chamber, and cell proliferation was assessed by MTT assay. The protein levels of RhoA, p-MLC (Thr18/Ser19) and alpha-SMA were assayed by Western blot.</p><p><b>RESULTS</b>Fasudil inhibited HSC adhesion, proliferation and LPA-induced migration in a concentration-dependent manner; the protein expressions of alpha-SMA and p-MLC (Thr18/Ser19) were significantly decreased in the presence of fasudil.</p><p><b>CONCLUSION</b>Fasudil can inhibit HSC adhesion, migration and proliferation by suppressing the cytoskeleton regulation function of Rho/ROCK signaling pathways.</p>

Changes of contents of extracellular signal-regulated kinase in rat hepatic fibrogenesis induced by bile duct ligation / 中国应用生理学杂志

<p><b>AIM</b>To observe the dynamic expression of ERK1 in fibrotic rat liver.</p><p><b>METHODS</b>The rat hepatic fibrosis was induced by bile duct ligation (BDL). Histopathological changes were evaluated by hematoxylin and eosin staining, and by Masson's trichrome method. ERK1 mRNA in liver was determined by reverse transcription-polymerase chain reaction (RT-PCR), while the distribution of ERK1 was assessed by immunohistochemistry. ERK1 protein was detected by using Western blotting analysis.</p><p><b>RESULTS</b>With the development of hepatic fibrosis, the positive cells of ERK1 increased a lot, they were mainly distributed at portal ducts, fiber septa and around the bile ducts, vascular endothelial cells and perisinusoidal cells. Western blotting analysis results displayed that the expression of ERK1 protein were up-regulated with model course, and its levels were the highest at week 4 after operation, achieving to 3.9-fold of that in normal rat liver. ERK1 mRNA expressed in normal rat livers as well, they were up-regulated at day 2 after BDL and its level was the highest at week 4 after BDL.</p><p><b>CONCLUSION</b>These data suggest that the expression of ERK1 and its mRNA can be increase greatly in fibrotic rat liver.</p>

The effect and mechanism of Salvia miltiorrhiza monomer IH764-3 on proliferation and collagen synthesis of hepatic stellate cells stimulated by H2O2 / 中国应用生理学杂志

<p><b>AIM</b>To investigate the inhibiting role of Salvia miltiorrhiza monomer IH764-3 in proliferation and collagen synthesis and the effect on the expression of focal adhesion kinase in hepatic stellate cells (HSCs) stimulated by H2O2 in vitro, so as to provide evidences for preventing hepatic fibrosis.</p><p><b>METHODS</b>The effects of IH764-3 on the proliferation and collagen synthesis of HSCs were examined by 3H-TdR and 3H-proline incorporation assay, respectively. The expression of FAK mRNA was examined by RT-PCR in lysates of cultured HSCs.</p><p><b>RESULTS</b>The proliferation and collagen synthesis were inhibited in dose-dependent and time-dependent manners. The expressions of FAK mRNA in the IH764-3 (10 microg/ml, 20 microg/ml, 30 microg/ml, 40 microg/ml) groups were significantly lower than that in the control group.</p><p><b>CONCLUSION</b>The present results demonstrate that IH764-3 can inhibit HSC proliferation and collagen synthesis. Meanwhile the mRNA transcription of FAK is reduced significantly, FAK as a signal molecule, can be involved in the effect of IH764-3. This may be one of the anti-fibrotic mechanisms of Salvia Miltiorrhiza.</p>

Effects of Arg-Gly-Asp-Ser tetrapeptide on integrin signaling and apoptosis in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effects of Arg-Gly-Asp-Ser (RGDS) tetrapeptide on integrin signaling and apoptosis in fibronectin (FN) -stimulated hepatic stellate cells (HSCs).</p><p><b>METHODS</b>3H-thymidine incorporation, annexin-V/propidium iodide double-labeled flow cytometry (FCM) and transmission electron microscopy were employed to estimate the influence of RGDS on the proliferation and apoptosis of HSCs. And the adhesion rates were observed by toluidine blue colorimetric assay. The expression of focal adhesion kinase (FAK) mRNA and protein in HSCs was detected using RT-PCR and western blotting analysis, respectively.</p><p><b>RESULTS</b>RGDS tetrapeptide at the concentrations of 25 microg/ml, 50 microg/ml and 100 microg/ml inhibited the proliferation of HSCs and induced HSCs apoptosis in dose-dependent and time-dependent manners, with the apoptotic rates of 9.49%, 27.67%, 31.59%, and the necrotic rates of 3.47%, 5.38%, 9.10%, respectively. Both the rates were higher than those in FN group (apoptotic rate: 3.44%; necrotic rate: 2.39%), F=8.02, P<0.05. After adding RGDS tetrapeptide to HSCs for 2 hours, the adhesive inhibition rates were 8.82%, 29.41% and 45.59%, respectively, which were higher than that in FN group (F=20.58, P<0.01). After exposure of HSCs to RGDS tetrapeptide for 24 hours, FAK protein decreased, and FAK mRNA was down-regulated earlier, about 2 hours after exposure to RGDS tetrapeptide.</p><p><b>CONCLUSION</b>These results suggest that RGDS tetrapeptide may induce apoptosis of HSC in both dose-dependent and time-dependent manners in vitro, which may be related to the disruption of cell matrix adhesion and down-regulation of FAK expression.</p>

Allograft transplantation with compound tissue of two ears and skin of head and neck: A report of 1 case, China / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the possibility and efficacy of allograft transplantation in treating patient with huge tissue defect after radical giant malignant melanoma resection.</p><p><b>METHODS</b>A male person received blood type matching was chosen as donor. Immediately after the donor's brain death, allograft was excised with the depth to the layer intervenient between periosteum and epicranial fascia in calvaria, the superficial layer of deep temporal fascia in both sides of temporal regions, close to zygomatic bones and mandibles including masseter and auricles upon in face, and cervical soft tissues including sternocleidomastoid muscles, cervical and external jugular vessels of both sides were excised simultaneously. After being perfused with 4 degrees C UW solution through both common carotid arteries, the homograft was sheared and radiated with X-ray before being preserved in UW solution for further use. During the operation, both sides of external auditory meatus were anastomosed with ears firstly, and vessels were anastomosed end-to-end sequently, at last, the border of skin flap was sutured intermittently. Combined use of MMF, FK506, Prednisone and Zenopax was performed as post-operation immunosuppressive treatment. Clinical observations were made on the signs and symptoms of graft survival or rejection as well as blood FK506 concentrations and immunological indexes were tested in laboratory. Biopsies of graft were also made at 1 h, 4 h, 8 h, 7 d, 14 d and 30 d after operation.</p><p><b>RESULTS</b>The circulation of the graft was satisfactory, and the temperature and color of skin were normal. Primary healing of suture and hair growth about 0.8 cm in a month were observed. Skin Biopsies of every time had no found of hyperacute or acute rejection. The concentration of FK506 was maintained 20 mg/ml 1 month after the operation.</p><p><b>CONCLUSION</b>Allograft transplantation with compound tissue of head skin flap and ears is a kind of effective and safe treatment in repairing huge tissue defect. Good tissue matching and combined use of currently available immunosuppressants can prevent hyperacute and acute rejection efficiently.</p>