Objective To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. Methods Twenty biopsies including scar tissues(n=8), chronic ulcer tissues(n=4) and normal tissues(n=8)were used in this study. The immunohistochemical staining (power visionTMtwo-step histostaining reagent) was used to explore the amount and expression characteristics of such protein.Results The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen ininflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. Conclusion Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing.
Hyperprolactinemia is the common clinical syndrome; the causes of hyperprolactinemia are physiological, pharmacological, and pathological, in which prolactinoma is the most common cause. In drug therapy, dopamine agonists are the first choice, but there are 10%–20% of the patients who are resistant to drug therapy. This paper mainly summarized the causes, treatments, mechanisms of drug resistance, treatment during pregnancy, and progresses in the treatment of prolactinoma, so as to provide some theoretical basis to further research of hyperprolactinemia.
Objective To investigate the expression of vascular endothelial growth factor-A （VEGF-A） and the phenotypic transition after the activation of fibroblasts by the supernatant of cultured tumor cells．Methods The growth tendency of fibroblasts was tested by the MTT assay．The expressions of alpha-smooth muscle actin （α-SMA） and VEGF-A mRNA were tested by RT-PCR．The expressions of α-SMA and VEGF-A protein were tested by immunohistochemistry and Western blot．Results The MTT assay indicated that the conditional medium which contained tumor cells supernatant could obviously promote the growth of the fibroblasts． RT-PCR and Western blot manifested that α-SMA expressed by the fibroblasts which cultured by normal medium reached its peak on day 5，then decreased to a low level on day 7．When the medium contained 2 ng/ml transforming growth factor-β1 （TGF-β1），the fibroblasts could steadily express more α-SMA．But the above two mediums could not make the fibroblasts express the VEGF-A. When using the conditional medium，the α-SMA peak advanced on the third day and maintained at a high level，so as the expression of the VEGF-A．Conclusions The results suggested that fibroblasts can be activated to be myofibroblasts when using the conditional medium．The best activation time of the fibroblasts is consistent with the time of the VEGF-A expression at the highest level by the activated fibroblasts．The fibroblasts which activated at the best time are expected to become a kind of cells which can be used for promoting revascularization．
ObjectiveTo investigate the mechanism of G protein coupled receptor kinase interacting protein 1 (GIT1) affecting angiogenesis by comparing the differentiation of bone marrow mesenchymal stem cells (BMSCs) differentiated into endothelial cells between GIT1 wild type mice and GIT1 gene knockout mice.MethodsMale and female GIT1 heterozygous mice were paired breeding, and the genotypic identification of newborn mice were detected by PCR. The 2nd generation BMSCs isolated from GIT1 wild type mice or GIT1 gene knockout mice were divided into 4 groups, including wild type control group (group A), wild type experimental group (group A1), GIT1 knockout control group (group B), and GIT1 knockout experimental group (group B1). The cells of groups A1 and B1 were cultured with the endothelial induction medium and the cells of groups A and B with normal cluture medium. The expressions of vascular endothelial growth factor receptor 2 (VEGFR-2), VEGFR-3, and phospho-VEGFR-2 (pVEGFR-2), and pVEGFR-3 proteins were detected by Western blot. The endothelial cell markers [von Willebrand factor (vWF), platelet-endothelial cell adhesion molecule 1 (PECAM-1), and vascular endothelial cadherin (VE-Cadherin)] were detected by flow cytometry. The 2nd generation BMSCs of GIT1 wild type mice were divided into 4 groups according to the different culture media: group Ⅰ, primary cell culture medium; group Ⅱ, cell culture medium containing SAR131675 (VEGFR-3 blocker); group Ⅲ, endothelial induction medium; group Ⅳ, endothelial induction medium containing SAR131675. The endothelial cell markers (vWF, PECAM-1, and VE-Cadherin) in 4 groups were also detected by flow cytometry.ResultsWestern blot results showed that there was no obviously difference in protein expressions of VEGFR-2 and pVEGFR-2 between groups; and the expressions of VEGFR-3 and pVEGFR-3 proteins in group A1 were obviously higher than those in groups A, B, and B1. The flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group A1 than in groups A, B, and B1 (P<0.05), and in group B1 than in groups A and B (P<0.05); but no significant difference was found between groups A and B (P>0.05). In the VEGFR-3 blocked experiment, the flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group Ⅲ than in groupsⅠ, Ⅱ, and Ⅳ, and in group Ⅳ than in groups Ⅰ and Ⅱ (P<0.05); but no significant difference was found between groups Ⅰ and Ⅱ (P>0.05).ConclusionGIT1 mediates BMSCs of mice differentiation into endothelial cells via VEGFR-3, thereby affecting the angiogenesis.
OBJECTIVE: To explore the expression of alpha-smooth muscle actin (alpha-SMA) induced by transforming growth factor beta 1 (TGF-beta 1). METHODS: Five samples of hypertrophic scars and three samples of normal mature scars were collected as the experimental and control groups respectively. The fibroblasts were isolated from scars, and cultured in 2-dimension or 3-dimension culture system. The immunohistochemical staining method of LSAB were used to investigate the expression of alpha-SMA in fibroblasts in the different concentration of TGF-beta 1. RESULTS: The expression of alpha-SMA in 3-dimension culture system were markedly lower than those in 2-dimension culture system with respect to the fibroblasts in the experimental group. The expression of alpha-SMA in fibroblasts were different in response to various TGF-beta 1 concentration, it was more effective at the concentration of 5 ng/ml. The expression of alpha-SMA in the fibroblasts from hypertrophic scars seemed to be more sensitive to TGF-beta 1 compared to that of the normal mature scars. CONCLUSION: There are concentration-dependent in the expression of alpha-SMA induced by TGF-beta 1 in scar fibroblasts in vitro. The biological characteristics of the fibroblasts from hypertrophic scars and normal mature scars and their sensitivity to the inducement of TGF-beta 1 were different. The inducement of TGF-beta 1 may be depressed by extracellular matrix components and that may decrease the expression of alpha-SMA.
Objective To explore the effect of cyclopamine （Cyc） which is the inhibitor of the Hedgehog signaling pathway on portal venous pressure of normal and liver cirrhosis rats, and it’s possible mechanisms. Moreover, to provide the experimental basis of drug efficacy and clinical treatment. Methods Thirty two healthy male SD rats were randomly average divided into four groups：normal control group, normal treatment group, liver cirrhosis control group, and liver cirrhosis treatment group. The liver cirrhosis models of rat were established by using the thioacetamide （TAA） method, which made 0.03% of TAA as the initial water concentration, and then the concentration of TAA in drinking water was adjusted according to the changes of the weekly body weight of rats lasting for twelve weeks. In thirteenth week, intraperitoneal injection of corn oil （0.1 ml/100 g body weight, 1 time/d） were performed lasting for a week in rats of the normal control group and liver cirrhosis control group; intraperitoneal injection of Cyc 〔1 mg （0.1 ml）/100 g body weight, 1 time/d〕were performed lasting for a week in rats of the normal treatment group and liver cirrhosis treatment group. In fourteenth week, the liver function, portal venous pressure （PVP）, and the ration of liver or spleen weight to body weight were detected, the expressions of α-smooth muscle actin （α-SMA） and typeⅠcollagen α1 （Col1α1） of hepatic stellate cell were detected by using immunohistochemistry. Results PVP were （10.7±0.9） and （12.3±1.3） cm H2O （1 cm H2O=0.098 kPa） in normal control group and normal treatment group, respectivly, the latter was higher than the former （t=-2.918，P=0.011）. PVP were （21.8±0.7） and （14.3±1.4） cm H2O in liver cirrhosis control group and liver cirrhosis treatment group, respectivly, the latter was lower than the former（t=13.602，P=0.000）. The expressions of α-SMA and Col1α1 in liver cirrhosis treatment group was lower than the liver cirrhosis control group. There were no significant difference of the liver function and ration of liver or spleen weight to body weight between the treatment group and the control group （P＞0.05）. Conclusion Cyclopamine could signally reduce the PVP of liver cirrhosis rats through reducing the expressions of α-SMA and Col1α1.
ObjectiveTo investigate the expression level and the clinical significance of Traf-2 and Nck-interacting protein kinase (TNIK) in gastric cancer tissues.MethodsWe retrospectively collected 78 cases of gastric cancer and its adjacent tissues which were diagnosed by surgery and pathology, all the patients received surgery in the Department of General Surgery in Luoyang Central Hospital Affiliated to Zhengzhou University, from July 2014 to December 2017, as well as 42 cases of gastric ulcer tissues in the same period. The expression of TNIK protein was detected by using immunohistochemical method and Western blot method in 78 cases of human gastric cancer tissues, corresponding adjacent normal tissues, and 42 cases of gastric ulcer tissues, the relationship between expression of TNIK protein and clinicopathologial features of gastric cancer was explored.ResultsThe positive expression rate of TNIK protein in gastric cancer tissues was 66.67% (52/78), and adjacent cancer tissues showed low expression rate of 11.54% (9/78), while it was 21.43% (9/42) in gastric ulcer tissues. On the positive expression rate of TNIK protein, gastric cancer tissues compared with adjacent cancer tissues and gastric ulcer tissues, the difference was statistically significant (P<0.05), however, there was no statistically significant difference between adjacent cancer tissues and gastric ulcer tissues (P>0.05). Western blot method showed that the expression level of TNIK protein in gastric cancer tissues was all significantly higher than that in adjacent cancer tissues and gastric ulcer tissues, the differences were statistically significant (P<0.05), but the difference was not significant when compared the adjacent cancer tissues and gastric ulcer tissues (P=0.772). The expression of TNIK protein was irrelevent to sexual distinction, CEA value, tumor diameter, and pathological type (P>0.05), while it was closely related to patients’ age, TNM stage, differentiation grade, distant metastasis, and lymph node metastasis (P<0.05). The prognosis of patients with positive expression of TNIK protein was worse than that of patients with negative expression of TNIK protein (P<0.05).ConclusionTNIK protein is highly expressed in gastric cancer tissues, which is associated with poorly prognostic factors and poor prognosis, and it may be a therapeutic target in patients with gastric cancer.
Objective To measure the expression level of Myc-interacting zinc finger protein-1 (MIZ1) in peripheral blood mononuclear cells (PBMC) of patients with severe and non-severe community-acquired pneumonia (CAP) and its relationship with inflammatory factors. Methods Thirty-six CAP patients from Beijing Chaoyang Hospital from April 2018 to June 2019 were enrolled in this study. MIZ1 mRNA level in PBMC were measured by reverse transcriptase-quantitative polymerase chain reaction. The levels of interleukin (IL)-6, IL-8, IL-10, and interferon-α in the serum of patients were measured by enzyme-linked immunosorbent assay. The levels of MIZ1 mRNA and inflammatory factors were compared between the severe CAP patients and the non-severe CAP patients. Results Compared with non-severe CAP patients, the MIZ1 mRNA level in the PBMC of severe CAP patients was lower (P<0.05) than non-severe group. Receiver operating characteristic (ROC) curve of the expression level of MIZ1 in PBMC was calculated according to whether CAP was severe or non-severe, and the area under ROC curve was 0.731 (P=0.018). Spearman correlation analysis showed that MIZ1 mRNA was negatively correlated with IL-10 level in the severe CAP patients (Spearman correlation co-efficient was –0.620, P<0.05). Conclusions MIZ1 may indicate the severity of CAP. MIZ1 may affect IL-10 so as to play a role in inflammation regulation.
Objective To validate the mechanism of effect of hepatic artery ischemia on biliary fibrosis after liver transplantation and the prevention method. Methods Eighteen male dogs were established into the concise auto orthotopic liver transplantation models and assigned into three groups randomly: hepatic artery ischemia (HAI) group, TBB group (transferred the blood by a bridge duct ) and control group, each group contained 6 dogs. After opening portal vein, the samples were cut from liver in each group at the time of 6 h, 3 d and 14 d. The pathological modifications of intrahepatic bile ducts were observed and expression of transforming growth factor-β1 (TGF-β1) were detected in the three times. Expressions of Smad3 and phosphate-Smad3 as well as mRNA of α-smooth muscle actin (α-SMA) in intrahepatic bile ducts were detected 14 d after opening portal vein.Results Compared with control group, the collagen deposition and lumens stenosis in biliary vessel wall were more obviously in HAI group. In TBB group, the pathological modifications were slighter compared with HAI group. The positive cell index of TGF-β1 reached peak on day 3 after opening portal vein, then decreased in TBB group, and which in HAI group kept increase and was significantly higher than that in TBB group （Plt;0.05）. The expression level of phosphate-Smad3 and transcriptional level of α-SMA mRNA were 1.04±0.13 and 1.12±0.55 in TBB group on day 14 after opening portal vein, which were significantly higher than those in control group (0.59±0.09 and 0.46±0.18) and lower than those in HAI group (1.82±0.18 and 1.86±0.73), the diversities among three groups were significant (Plt;0.05). There was not significant difference of expression of Smads among three groups (Pgt;0.05). Conclusions Hepatic artery ischemia could increase the deposition of collagen fibers and the transdifferentiation of myofibroblast in bile duct and result in the biliary fibrosis by activating the TGF-β1/Smads signaling pathway. The bridging bypass device could lessen the biliary fibrosis caused by hepatic artery ischemia by inhibiting the activation of TGF-β1/Smads signal transduction passageway.
Objective To investigate the modulating effect of transforming growth factor beta;2 (TGFbeta;2) and extracellular matrix (ECM) on the transdifferentiation of human fetal RPE (hfRPE) cells into myofibroblast-like cells , and to determine the mechanism of signal transduction. Methods hfRPE cells were cultured on ECM coated or uncoated petri dish with or witho ut TGFbeta;2 in the medium. The expression of alpha;-smooth muscle actin (alpha;-SMA) were detected by immunocytochemistry examination, flow cytometry and Western blotting via calphostin C, genistein, PD98059, and Wortmannin. Results After cultured on ECM coated petri dish with TGFbeta;2 in the medium,there were obvious morphological changes of hfRPE cells including cellular elongating and appearing of actin microfilaments. The results of flow cytometry and immunocytochemistry examination showed that expression of alpha;-SMA obviously increased after TGFbeta;2 was added in the medium in a dose-dependent manner. Compared with which of hfRPE cells cultured on the uncoated surface of culture plates, the total mean fluore scence intensity (TMFI) of hfRPE cells cultured on FN-coated surface increased (38.01plusmn;1.14)% when the stimulation concentration of TGFbeta;2 was 50ng/ml(Plt;0.05). Western blotting further confirmed the effects. The changes mentioned above could be inhibited mostly by protein kinase C (PKC) and calphostin C (10 nmol/L)(Plt;0.01). Conclusion TGFbeta;2 may induce the transdifferentiation of hfRPE cells into myofibroblast-like cells in a dose dependent manner, which could be intensified by FN. These mediated effects of TGFbeta;2 and ECM may act via the PKC signal transduction pathway. (Chin J Ocul Fundus Dis, 2006, 22: 328-332)