Effect of 6-Shogaol on invasion and migration of triple negative breast cancer cells via Hedgehog/Glil signaling pathway / 中国药理学通报

Aim To explore the effects of the expression of the transcription faetor Glil of Hedgehog ( Hh ) signaling pathway and the 6-Shogaol mediated Hedge- hog/Glil pathway on the proliferation, invasion and migration in MDA-MB-231 eells of triple negative breast eaneer.Methods MDA-MB-231 eells were transfected by lentiviral vectors to stably overexpressed Glil gene.The overexpression efficiency of Glil was verified by qRT-PCR and Western blot.CCK-8 and EdlJ assays were used to detect the effect of Glil expression and 6-Shogaol on cell viability.Cell scratch assay and Transwell assay were used to detect the ability of migration and invasion.Western blot was used to detect the proteins expression of Hedgehog signaling pathway and other related genes.Results MDA-MB- 231-Glil overexpression cell line was successfully established.When Gli 1 gene was overexpressed, the invasion and migration ability of cells was significantly improved, anrl the expression of Hh signaling pathway gene Glil , EMT marker gene Vimentin, Hippo signaling pathway genes YAP and TEAD4 inereased.When the expression of Glil was inhibited by the Hh/Gli pathway inhibitor Gant61 , the proliferation, invasion and migration abilities were suppressed.When the eells were treated with 6-Shogaol, the abilities of proliferation, invasion and migration were inhibited as well as the proteins expression of Glil , Vimentin, YAP and TEAD4 deereased.Conclusions Glil gene ean promote the invasion and migration of MDA-MB-231 eells.6-Shogaol ean inhibit proliferation, invasion and migration of breast eaneer eells through Hedgehog signaling pathway, suggesting that transcription factor Glil may be one of the targets of 6-Shogaol.

A Biomechanical Analysis of the Interlock Suture and a Modified Kessler-Loop Lock Flexor Tendon Suture

OBJECTIVE: In this work, we attempted to develop a modified single-knot Kessler-loop lock suture technique and compare the biomechanical properties associated with this single-knot suture technique with those associated with the conventional modified Kessler and interlock suture techniques. METHODS: In this experiment, a total of 18 porcine flexor digitorum profundus tendons were harvested and randomly divided into three groups. The tendons were transected and then repaired using three different techniques, including modified Kessler suture with peritendinous suture, interlock suture with peritendinous suture, and modified Kessler-loop lock suture with peritendinous suture. Times required for suturing were recorded and compared among groups. The groups were also compared with respect to 2-mm gap load, ultimate failure load, and gap at failure. RESULTS: For tendon repair, compared with the conventional modified Kessler suture technique, the interlock and modified Kessler-loop lock suture techniques resulted in significantly improved biomechanical properties. However, there were no significant differences between the interlock and modified Kessler-loop lock techniques with respect to biomechanical properties, gap at failure, and time required. CONCLUSIONS: The interlock and modified Kessler-loop lock techniques for flexor tendon sutures produce similar mechanical characteristics in vitro.

Prognosis and Survival Analysis of Patients with Essential Thrombocythemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the survival status and prognosis of patients with essential thrombocythemia(ET) and analyze the prognostic factors for the patients' survival, so as to provide a evidence for clinical treatment and prognosis evaluation.</p><p><b>METHODS</b>A retrospective analysis of 118 patients with ET was conducted in the Fifth Affiliated Hospital of Sun Yat-Sen University and Zhongshan Municipale People's Hospital from December 2002 to December 2013. The clinical characteristics were summarized, such as the survival curve and multi-factor analysis, therefore looking for the disease characteristics and risk factors affecting the survival and prognosis.</p><p><b>RESULTS</b>Among 118 ET patients enrolled in this study, the survival rate of ET patients for 1, 3, 5 and 10 years were 95.5%,92.6%,89% and 81.6%, respectively. Kaplan-Meier survival curve showed that the age ≥60 years old at diagnosis, cardiovascular risk factors, anamnesis of thrombosis or hemorrhage, anemia(hemoglobin<120 g/L), thrombocythemia (≥1 000×10/L), risk stratification and hydroxyurea or HHT(hemoharringtonine) use in high-risk group were factors affecting the suvival rate, 7 out of those factors influencing survival rate were statistically significant (P<0.05). COX regression analysis showed that independent risk factors affecting survival have not yet been found.</p><p><b>CONCLUSION</b>ET patients display a high survival rate and long survival time, and their conversion risk into the marrow fibrosis or leukemia has been found to be low. The age≥60 years old at diagnosis, cardiovascular risk factors, anamnesis of thrombosis or hemorrhage, anemia and therombocythemia are the risk factors affecting prognosis. The use of hydroxyurea or HHT in high-risk group can improve the prognosis.</p>

Peroxisome proliferator-activated receptor-γ agonist pioglitazone fails to attenuate renal fibrosis caused by unilateral ureteral obstruction in mice / 华中科技大学学报（医学）（英德文版）

Renal tubulointerstitial fibrosis is the common ending of progressive renal disease. It is worth developing new ways to stop the progress of renal fibrosis. Peroxisome proliferator-activated receptor-γ (PPARγ) agonists have been studied to treat diabetic nephropathy, cisplatin-induced acute renal injury, ischemia reperfusion injury and adriamycin nephropathy. In this study, unilateral ureteral obstruction (UUO) was used to establish a different renal fibrosis model. PPAR? agonist pioglitazone was administrated by oral gavage and saline was used as control. At 7th and 14th day after the operation, mice were sacrificed for fibrosis test and T lymphocytes subsets test. Unexpectedly, through MASSON staining, immunohistochemistry for α-SMA, and Western blotting for a-SMA and PDGFR-β, we found that pioglitazone failed to attenuate renal fibrosis in UUO mice. However, flow cytometry showed that pioglitazone down-regulated Th1 cells, and up-regulated Th2 cells, Th17 cells and Treg cells. But the Th17/Treg ratio had no significant change by pioglitazone. Real-time PCR results showed that TGF-β and MCP-1 had no significant changes, at the same time, CD4(+) T cells associated cytokines were partially regulated by pioglitazone pretreatment. Taken together, pioglitazone failed to suppress renal fibrosis progression caused by UUO.

Peroxisome proliferator-activated receptor-γ agonist pioglitazone fails to attenuate renal fibrosis caused by unilateral ureteral obstruction in mice / 华中科技大学学报（医学）（英德文版）

Renal tubulointerstitial fibrosis is the common ending of progreβsive renal disease. It is worth developing new ways to stop the progreβs of renal fibrosis. Peroxisome proliferator-activated receptor-γ (PPARγ) agonists have been studied to treat diabetic nephropathy, cisplatin-induced acute renal injury, ischemia reperfusion injury and adriamycin nephropathy. In this study, unilateral ureteral obstruction (UUO) was used to establish a different renal fibrosis model. PPAR? agonist pioglitazone was administrated by oral gavage and saline was used as control. At 7th and 14th day after the operation, mice were sacrificed for fibrosis test and T lymphocytes subsets test. Unexpectedly, through MASSON staining, immunohistochemistry for α-SMA, and Western blotting for a-SMA and PDGFR-β, we found that pioglitazone failed to attenuate renal fibrosis in UUO mice. However, flow cytometry showed that pioglitazone down-regulated Th1 cells, and up-regulated Th2 cells, Th17 cells and Treg cells. But the Th17/Treg ratio had no significant change by pioglitazone. Real-time PCR results showed that TGF-β and MCP-1 had no significant changes, at the same time, CD4(+) T cells associated cytokines were partially regulated by pioglitazone pretreatment. Taken together, pioglitazone failed to suppress renal fibrosis progression caused by UUO.

Erbin interacts with Sema4C and inhibits Sema4C-induced epithelial-mesenchymal transition in HK2 cells / 华中科技大学学报（医学）（英德文版）

Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition (EMT) in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the interaction between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and (or) Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. After HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.

Erbin interacts with Sema4C and inhibits Sema4C-induced epithelial-mesenchymal transition in HK2 cells / 华中科技大学学报（医学）（英德文版）

Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition (EMT) in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the interaction between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and (or) Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. After HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.

Rapid differentiation of human umbilical cord-derived mesenchymal stem cells into insulin-secreting cells under the sole induction of biological products / 生理学报

In order to explore the feasibility of inducing the human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) to differentiate into insulin-secreting cells with biological products alone, hUC-MSCs were separated and purified from the whole umbilical cord by the sequent digestion of collagenase II and trypsin followed by two-step centrifugation. hUC-MSCs were induced with IMDM culture medium containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and Ginkgo biloba extract (GBE). Before and after the induction, the morphological changes were observed under inverse microscope; the islet-related genes were detected by RT-PCR; islet-like clusters (ILCs) were identified by dithizone (DTZ) staining; PDX-1 and immunoreactive insulin (IRI) were examined by immunofluorescence method; the quantity and quality of IRI secretion were assayed by chemiluminescence immunoassay and Western blot respectively. The results showed that the purified hUC-MSCs presented long spindle-like shape and parallel or spiral arrangement which are typical morphological features of MSCs. After the induction, hUC-MSCs changed gradually into round or oval shape and gathered together to form ILCs; there were more than one hundred clusters on the growth surface of a flask of T25; ILCs were stained into positive mauve by DTZ and positive for PDX-1 and IRI; Western blot displayed that most of the IRI was proinsulin (PI). Therefore, hUC-MSCs can rapidly differentiate into insulin-secreting cells under the sole induction of EGF, bFGF, GBE and IMDM, but ILCs are not mature enough to produce sufficient true insulin.

Differentiation of hepatic oval cell into mature hepatocyte induced by hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of hepatic stellate cells in the differentiation of hepatic oval cells into adult hepatocyte.</p><p><b>METHODS</b>The oval cell were cocultured with primary hepatic stellate cells (HSC) in the same well (M-coculture) or separately cultured with HSC by millIcell (S-coculture). Oval cells were cultured alone as control; the expression of adult hepatocyte marker HNF-4alpha, albumin, and oval cell marker AFP, CK-19 in each group were detected by real-time PCR and western-blot. Phenotype changes were observed by transmission electron microscope (TEM); PAS staining was used to detect the quantity of glycogen granule in oval cell. Albumin level in supernatant was detected using ELISA kit.</p><p><b>RESULT</b>(1) The relative level of HNF-4alpha and albumin mRNA expression compared with pre-coculture: M-coculture: HNF-4a: 1.9+/-0.2, 10.7+/-1.2, 12.0+/-1.3; albumin: 5.7+/-1.6, 110.7+/-13.7, 173.6+/-22.3. S-coculture: 1.4+/-0.1, 3.2+/-0.6, 8.9+/-1.4 times; albumin: 2.9+/-1.4, 22.3+/-8.5, 96.3+/-16.3. The relative level of HNF-4a and albumin mRNA expression in coculture group (M- and S-coculture) were higher than control group (LSD-t: 32.98, 10.08, 13.38, 7.96; P less than 0.01); and a higher level of HNF-4a and albumin was found in M-coculture group compared to S-coculture group (LSD-t: 32.98, 25.65; P less than 0.01). The relative level of AFP and CK-19 mRNA expression compared with pre-coculture: M-coculture: 1.1+/-0.2, 0.2+/-0.0, 0.0+/-0.0; S-coculture group: AFP: 1.0+/-0.2, 0.2+/-0.1, 0.1+/-0.0; CK-19: 0.6+/-0.1, 0.1+/-0.0, 0.0+/-0.0; control group: AFP: 1.0+/-0.1, 1.0+/-0.1, 1.1+/-0.1, CK-19: 1.0+/-0.1, 1.1+/-0.1, 1.0+/-0.1. The relative level of AFP and CK-19 mRNA expression in coculture group (M- and S-coculture) were lower than that in control group (LSD-t: 37.99, 34.50, 13.59, 22.46; P less than 0.01). (2) The albumin secretion was detected in M-coculture: 14 day: (15.30+/-0.09) ng/ml, 21: (20.98+/-0.12) ng/ml; S-coculture: 14 day: (11.41+/-0.13) ng/ml, 21 day:(15.12+/-0.17) ng/ml. (3) It showed more organelles such as endoplasmic reticulum, mitochondrion and Golgi apparatus in oval cells cocultured with HSC. And cholangiole-like structure appeared between oval cells cocultured with HSC. (4) PAS staining showed glycogen granules could be observed in coculture groups.</p><p><b>CONCLUSION</b>HSC can induce differentiation of oval cell into mature hepatocyte.</p>