Value of autotaxin in predicting refractory Mycoplasma pneumoniae pneumonia in children and its correlation with inflammatory cytokines / 中国当代儿科杂志

OBJECTIVES@#To study the value of autotaxin (an autocrine motility factor) level in serum and bronchoalveolar lavage fluid (BALF) in predicting refractory Mycoplasma pneumoniae pneumonia (RMPP) in children and its correlation with interleukin-6 (IL-6), interleukin-8 (IL-8), and C-reactive protein (CRP).@*METHODS@#A retrospective analysis was performed on 238 children with Mycoplasma pneumoniae pneumonia who were admitted from January 2019 to December 2021. According to disease severity, they were divided into two groups: RMPP (n=82) and general Mycoplasma pneumoniae pneumonia (GMPP; n=156). The two groups were compared in terms of the levels of autotaxin, IL-6, IL-8, and CRP in serum and BALF to study the value of autotaxin level in serum and BALF in predicting RMPP in children, as well as the correlation of autotaxin level with IL-6, IL-8, and CRP in children with RMPP.@*RESULTS@#Compared with the GMPP group, the RMPP group had significantly higher levels of autotaxin, IL-6, IL-8, and CRP in serum and BALF (P<0.05). For the children with RMPP, the levels of autotaxin, IL-6, IL-8, and CRP in serum and BALF in the acute stage were significantly higher than those in the convalescent stage (P<0.05). The receiver operating characteristic (ROC) curve showed that the level of autotaxin in serum and BALF had a good value in predicting RMPP in children, with an area under the curve of 0.874 (95%CI: 0.816-0.935) and 0.862 (95%CI: 0.802-0.924), respectively. The correlation analysis showed that the level of autotaxin in serum and BALF was positively correlated with IL-6, IL-8, and CRP levels (P<0.001).@*CONCLUSIONS@#The level of autotaxin in serum and BALF increases and is correlated with the degree of disease recovery and inflammatory cytokines in children with RMPP. Autotaxin can be used as a predictive indicator for RMPP in children.

Study of association between expression of E-cadherin and Autocrine Motility Factor in different grades of Colorectal adenocarcinoma

Though incidence of Colorectal carcinoma is relatively less in consideration of world wide data, Indian scenario differs in higher incidence of Grade 3 carcinoma with signet ring cell component. We tried to find association between intercellular adhesion and propagation of cancer cell in Adenocarcinoma of Colorectal region. E-cadherin (ECAD) is the strongest intercellular adhesion molecule of epithelial cells and Autocrine Motility Factor (AMFR) is known propagator of cancer cell. We studied simultaneous immunohistochemical expression of these two molecules in 92 already diagnosed cases that were treated surgically by Colectomy or Abdomino Pelvic Resection. Normal Colorectal mucosa reacted strongly with ECAD and weakly with AMFR. With increasing dedifferentiation of the tumor, reversal manifested- more aggressive grades showed varied expression in comparison to its less aggressive type. Gastric 3 adenocarcinoma show weak ECAD (83% vs. 35%) and more AMFR (93% vs. 35%) expression in comparison to Grade 1. Weak ECAD and strong AMFR was also associated with increase in depth of tumor invasion. As ECAD and AMFR is at least partially responsible for varying histologic grade and behavioral pattern of colorectal adenocarcinoma, simultaneous evaluation of both parameters is helpful to understand pathway of progression of such cancer cell.

The treatment effect of novel hGHRH homodimer to male infertility hamster

Extra-hypothalamic growth hormone-releasing hormone (GHRH) plays an important role in reproduction. To study the treatment effect of Grin (a novel hGHRH homodimer), the infertility models of 85 male Chinese hamsters were established by intraperitoneally injecting 20 mg/kg of cyclophosphamide once in a week for 5 weeks and the treatment with Grin or human menopausal gonadotropin (hMG) as positive control was evaluated by performing a 3-week mating experiment. 2–8 mg/kg of Grin and 200 U/kg of hMG showed similar effect and different pathological characteristics. Compared to the single cyclophosphamide group (0%), the pregnancy rates (H-, M-, L-Grin 26.7, 30.8, 31.3%, and hMG 31.3%) showed significant difference, but there was no difference between the hMG and Grin groups. The single cyclophosphamide group presented loose tubules with pathologic vacuoles and significant TUNEL positive cells. Grin induced less weight of body or testis, compactly aligned tubules with little intra-lumens, whereas hMG caused more weight of body or testis, enlarging tubules with annular clearance. Grin presented a dose-dependent manner or cell differentiation-dependentincrease in testicular GHRH receptor, and did not impact the levels of blood and testicular GH, testosterone. Grin promotes fertility by proliferating and differentiating primitive cells through up-regulating testicular GHRH receptor without triggering GH secretion, which might solve the etiology of oligoasthenozoospermia.

Study of association between expression of E-cadherin and Autocrine Motility Factor in different grades of Colorectal adenocarcinoma

Though incidence of Colorectal carcinoma is relatively less in consideration of world wide data, Indian scenario differs in higher incidence of Grade 3 carcinoma with signet ring cell component. We tried to find association between intercellular adhesion and propagation of cancer cell in Adenocarcinoma of Colorectal region. E-cadherin (ECAD) is the strongest intercellular adhesion molecule of epithelial cells and Autocrine Motility Factor (AMFR) is known propagator of cancer cell. We studied simultaneous immunohistochemical expression of these two molecules in 92 already diagnosed cases that were treated surgically by Colectomy or Abdomino Pelvic Resection. Normal Colorectal mucosa reacted strongly with ECAD and weakly with AMFR. With increasing dedifferentiation of the tumor, reversal manifested- more aggressive grades showed varied expression in comparison to its less aggressive type. Gastric3 adenocarcinoma show weak ECAD (83% vs. 35%) and more AMFR (93.% vs. 35%) expression in comparison to Grade1.Weak ECAD and strong AMFR was also associated with increase in depth of tumor invasion. As ECAD and AMFR is at least partially responsible for varying histologic grade and behavioral pattern of colorectal adenocarcinoma, simultaneous evaluation of both parameters is helpful to understand pathway of progression of such cancer cell.

Autocrine IGF-1/IGF-1R signaling promotes cell migration and invasion in NK/T-cell lymphoma cells / 白血病·淋巴瘤

Objective To identify the expression pattern of IGF-1 and IGF-1R in NK/T-cell lymphoma (NK/TCL) cell lines and to investigate the role of IGF-1/IGF-1R signaling in regulation of cell migration and invasion.Methods RT-PCR and immunofluorescence were performed to detect the expression of IGF-1 and IGF-1R.Transwell assay was applied to observe the effects of IGF-1/IGF-1R signaling and downstream kinases activities on cell migration and invasion.Concentrations of MMP-2 and MMP-9 were quantified by ELISA.Results Co-expression of IGF-1 and its receptor IGF-1R were identified in two NK/TCL cell lines,SNK-1 and SNK-6,while normal NK cells lack the IGF-1R expression.IGF-1R inhibitors significantly reduced SNK-1 and SNK-6 cells migration and invasion rates.Exogenous IGF-1 promoted both cell lines migration and invasion,but these effects were both blocked by IGF-1R inhibitors.Inhibition of AKT,p38 and JNK,the possible IGF-1R downstream kinases,reduced cell migration rates.Further more,exogenous IGF-1 significantly increased MMP-2 and MMP-9 secretion,while decreased secretion of MMP-2 and MMP-9 were observed when IGF-1R inhibitors were applied.Conclusion An autocrine IGF-1/IGF-1R signaling loop is aberrantly expressed on NK/TCL cells and the autocrine loop significantly promotes cell migration and invasion through activation of p38,PI3K and JNK signaling and enhances secretion of MMP-2 and MMP-9.

Effect of down-regulation of phosphoglucose isomerase/autocrine motility factor gene with ginsenoside Rh2 on leukemia KG1α cells in vitro / 中草药

Objective: To investigate the down-regulation of phosphoglucose isomerase/autocrine motility factor (PGI/AMF) gene expression by interfering RNA for study on the pharmacological effect of PGI/AMF with ginsenoside Rh2 on leukemia KG1α cells. Methods: The KG1α and the silencing PGI/AMFα KG1α (siPGI-KG1α) cells at logarithmic growth phase were divided into control and drug groups in different dosages. The cells in the control group were normally treated and the cells in the drug groups were incubated with ginsenoside Rh2 in the concentration of 30, 45, 60, 75, and 90 μmol/L in 24, 48, and 72 h respectively. Cell Counting Kit-8 (CCK-8) was used to demonstrate the effect of ginsenside Rh2 on proliferation of KG1α and siPGI-KG1α cells. Trypan blue staining was applied to detecting the effect of human PGI on the proliferation of leukemia KG1α cell. The changes of KG1α by ginsenoside Rh2 on Akt pathway were tested by Antibody Array. The corelation of down-regulating PGI/AMF and Rh2 about mTOR, Raptor, and Rag was detected by Western blotting. Results: Compared with the control group, ginsenoside Rh2 had significant inhibition on the proliferation of KG1α. Down-regulation of PGI/AMF could make KG1α more sensitive to ginsenoside Rh2 and down-regulation of PGI could inhibit the proliferation of KG1α. Antibody Array showed that ginsenoside Rh2 could inhibit the proliferation of KG1α cells through decreasing the expression of P38, mTOR, Akt, AMPKα, PARP, and Bad. Western blotting results indicated that down-regulation of PGI through the synergy of mTOR, Rag, and Raptor with ginsenoside Rh2 could inhibit the proliferation of KG1α cells. Conclusion: Down-regulation of PGI/AMF coordinated with ginsenoside Rh2 could inhibit the proliferation of KG1α cells.

The relationships between the expression of autocrine motility factor receptor and the clinical features of papillary thyroid carcinoma / 中国医师杂志

ObjectiveTo investigate the expression of autocrine motility factor receptor (AMFR)in papillary thyroid carcinoma and its relationship with clinical characteristics of this disease.Methods Real - time quantitative PCR and immunohistochemical methods were used to analyze the expression of AMFR in papillary thyroid carcinomas.ResultsThe significant differences in AMFR expression between papillary thyroid carcinoma and normal thyroid tissues were found in the levels of mRNA (6.296 ± 1.568 vs 7.913 ± 2.351,t=3.681,P=0.001 ) and protein ( 63.1％ vs 34.5 ％,x2=13.722,P ＜ 0.001 ),respectively.Immunohistochemistry analyses showed that the protein expression of AMFR in papillary thyroid carcinomas were significantly correlated with tumour size ( x2=5.209,P ＜ 0.05 ) and lymph node metastasis ( x2=4.32,P ＜ 0.05 ),and it was affected by the factors age ( x2=0.739,P=0.39 ) and gender ( x2=0.064,P=0.81 ).ConclusionsThe increased AMFR in papillary thyroid carcinoma would be a new target for cancer therapy and a new marker for prognosis.

Epigallocatechin-3-gallate inhibits paracrine and autocrine hepatocyte growth factor/scatter factor-induced tumor cell migration and invasion

Aberrant activation of hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, Met, is involved in the development and progression of many human cancers. In the cell-based screening assay, (-)epigallocatechin-3-gallate (EGCG) inhibited HGF/SF-Met signaling as indicated by its inhibitory activity on HGF/SF-induced cell scattering and uPA activation (IC50 = 15.8 microg/ml). Further analysis revealed that EGCG at low doses specifically inhibited HGF/SF-induced tyrosine phosphorylation of Met but not epidermal growth factor (EGF)-induced phosphorylation of EGF receptor (EGFR). On the other hand, high-dose EGCG decreased both Met and EGFR proteins. We also found that EGCG did not act on the intracellular portion of Met receptor tyrosine kinase, i.e., it inhibited InlB-dependent activation of Met but not NGF-induced activation of Trk-Met hybrid receptor. This inhibition decreased HGF-induced migration and invasion by parental or HGF/SF-transfected B16F10 melanoma cells in vitro in either a paracrine or autocrine manner. Furthermore, EGCG inhibited the invasion/metastasis of HGF/SF-transfected B16F10 melanoma cells in mice. Our data suggest the possible use of EGCG in human cancers associated with dysregulated paracrine or autocrine HGF/SF-Met signaling.

Maintenance of undifferentiated state of human embryonic stem cells in chemical defined medium at high clone density without exogenous cell factors / 中南大学学报(医学版)

Objective To establish human embryonic stem cells (hESCs) feeder-independent and cell factor-free culture system. Methods Effect of high and low clone densities of hESCs culture was compared and impact of the clone densities to hESCs culture media was analyzed. Results HESCs could maintain their undifferentiated states at high clone density (34 clones/cm2) without cell factors. At the same time,the bone morphology protein (BMP)-like induction of N2 and B27 supplements (NB) medium could be modulated by the clone density,and high level of BMP-like induction was accompanied by high clone density. Conclusion High clone density of hESCs can change the environments by themselves to maintain the undifferentiated states,which provides a new clue to explore the mechanism of undifferentiated states of hESCs and simplify the culture medium.

Research progress in the relationship of autocrine motility factor,autocrine motility factor receptor and the nervous system / 中国药理学通报

Autocrine motility factor (AMF) plays an important role in the stimulation of the migration and motility of cells, especially the generation, migration and angiogenesis of tumor. Recently, it has been found that AMF has three isoforms, ATX-t, ATX-m and PD-I alpha. The PD-I alpha isoform is specifically expressed in the brain, which plays extensive functions in nervous system, such as regulating neural development and differentiation, promoting neurotrauma repair, inducing neuropathic pain, even contributing neurodegeneration under some circumstances. This indicates the close relationship of AMF/AMFR and the pathophysiology of the nervous system. This paper mainly reviews the function of AMF and AMFR and its possible mechanism in the nervous system.

Transfection of rat myoblasts with leuflvirus carrying autocrine motility factor gene / 中华创伤杂志

Objective To explore a safe and high efficiency way of gene transfection of autocrine motility factor(AMF) in order to provide experimental basis for transplantation of myoblasts carrying AMF gone. Methods Sprague Dawley rat myoblasts were cultured, purified, proliferated and immunohisto-chemically verified. Then, the myoblasts were transfected with AMF and eGFP (enhanced green fluores-cent protein) gene by FIV (feline immunodeficiency virus). Fluorescence microscope and laser scanning confocal microscope were employed to detect eGFP so as to verify positive transfection rate. Expression of AMF was detected by immunohistochemical method. Results Myoblasts with 98% purity could he ob-tained after two weeks of primary culture and purification. Positive transfection rate reached 90.4% when MOI (multiplicity of infection) was 100 (P <0.01). The transfected AMF gene could express normally. Conclusions Explant culture is a proper way in rat myoblast culture. Meanwhile, AMF gene can he effectively transfected into rat myoblast and well expressed via FIV.

VEGF-related Autocrine Growth in Periosteal-Derived Cells

PURPOSE: The development of a microvascularization is important for the homeostasis of normal bone. Vascular endothelial growth factor (VEGF) is one of the most important factors in vessel formation. The purpose of this study was to examine VEGF-related autocrine growth in periostealderived cells. MATERIALS AND METHODS: Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured for 21 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. RESULTS: The expression of four VEGF isoforms and VEGFRs was observed in periosteal-derived cells. Treatment with cultures with VEGFR-1 and VEGFR-2 Kinase Inhibitor inhibited osteoblastic differentiation and alkaline phosphatase (ALP) activity of periosteal-derived cells. In addition, exogenous VEGF treatment increased calcium content in the periosteal-derived cells. CONCLUSION: These results suggest that VEGF might act as an autocrine growth molecule during osteoblastic differentiation of cultured human periosteal-derived cells.

VEGF-related Autocrine Growth in Periosteal-Derived Cells

PURPOSE: The development of a microvascularization is important for the homeostasis of normal bone. Vascular endothelial growth factor (VEGF) is one of the most important factors in vessel formation. The purpose of this study was to examine VEGF-related autocrine growth in periostealderived cells. MATERIALS AND METHODS: Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured for 21 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. RESULTS: The expression of four VEGF isoforms and VEGFRs was observed in periosteal-derived cells. Treatment with cultures with VEGFR-1 and VEGFR-2 Kinase Inhibitor inhibited osteoblastic differentiation and alkaline phosphatase (ALP) activity of periosteal-derived cells. In addition, exogenous VEGF treatment increased calcium content in the periosteal-derived cells. CONCLUSION: These results suggest that VEGF might act as an autocrine growth molecule during osteoblastic differentiation of cultured human periosteal-derived cells.

Regulation of expression of matrix metalloproteinase-9 by JNK in Raw 264.7 cells: presence of inhibitory factor(s) suppressing MMP-9 induction in serum and conditioned media

Matrix metalloproteinase-9 (MMP-9) secreted from macrophages plays an important role in tissue destruction and inflammation through degradation of matrix proteins and proteolytic activation of cytokines/chemokines. Whereas the MEK-ERK and PI3K-Akt pathways up-regulate MMP-9 expression, regulation of MMP-9 by JNK remains controversial. Presently, we aimed to determine the role of JNK in MMP-9 regulation in Raw 264.7 cells. Inhibition of JNK by the JNK inhibitor SP600125 induced MMP-9 in the absence of serum and suppressed the expression of TNF-alpha, IL-6 and cyclooxygenase-2 in LPS-treated Raw 264.7 cells. In a knockdown experiment with small interfering RNA, suppression of JNK1 induced MMP-9 expression. Interestingly, mouse serum suppressed SP600125-mediated MMP-9 induction, similar to IFN-gamma. However, the inhibitory activity of mouse serum was not affected by pyridone 6, which inhibits Janus kinase downstream to IFN-gamma. In addition to mouse serum, conditioned media of Raw 264.7 cells contained the inhibitory factor(s) larger than 10 kDa, which suppressed SP600125- or LPS-induced MMP-9 expression. Taken together, these data suggest that JNK1 suppresses MMP-9 expression in the absence of serum. In addition, the inhibitory factor(s) present in serum or secreted from macrophages may negatively control MMP-9 expression.

Evaluation of angiogenic phenotypes in cultured human periosteal-derived cells under high-dose dexamethasone

Exogenous substance P induced autocrine of substance P in epidermal stem cells / 中华创伤杂志

Objective To study autocrine characteristics of substance P(SP)in epidermal stem cells(ESCs). Methods In vitro cultured ESCs was stimulated by exogenous SP.Then,the expressions of SP gene,protein and exogenous SP protein in ESCs and the supernatant fluid were detected by immunofluorescence,ELISA,Western blot and real-time PCR. Results After stimulation by exogenous SP,the expression of SP protein in ESCs and supernatant fluid was up-regulated and reached peak at 24th hour,with higher concentration than the added SP.The gene expression of SP was up-regulated to the peak 6 hours after stimulation by SP.Both expression of gene and protein remained unchanged after the natural killer cell receptor Was blocked even by stimulation with exogenous SP. Conclusion Anticrine of SP of ESCs can be induced by exogenous SP.

Combined assays of tumor marker CEA and TSGF for differential diagnosis of pleural effusion / 中国基层医药

Objective To study the clinical significance of tumor markers combined assay in the differential diagnosis of pleural effusion.Methods 165 serum and hydrothorax samples were collected from 165 in-patients with pleural effusion.Among them ,55 patients were suffered from malignant pleural effusion and 110 patients with benign pleural effusion.The concentration of CEA,and TSGF were determined by chemiluminescence assay.Results In malignant pleural effusion group,the positive rates of serum CEA and TSGF were 58.8% and 54.3% respectively,the positive rates of hydrothorax CEA and TSGF were 75.8% ,70.7% respectively.Among them,CEA and TSGF in malignant group were significantly higher than that in benign pleural effusion group(P < 0.01 ).So as did of the concentration.The sensitivity of combined assay with serum TSGF in diagnosing malignant pleural effusion was 85.7% ,with the specificity of 97.8%.Conclusion Tumor markers of CEA and TSGF have high clinical values in the differential diagnosis of pleural effusion.The combined assay with TSGF is recommended.

An experimental study for the detection of autocrine growth activity in the osteogenic cells after mandibular distraction osteogenesis

BACKGROUND: Distraction osteogenesis (DO) is a useful method for treating cases demanding the generation of new bone. During DO, the angiogenic activity is crucial factor in the new bone formation. The aim of this study was to detect the autocrine growth activity in the cellular components of the distracted bone with observation of the co-expression of vascular endothelial growth factor (VEGF) and its receptors following the mandibular DO. MATERIALS AND METHODS: Unilateral mandibular distraction (0.5 mm twice per day for 10 days) was performed in six mongrel dogs. Two animals were killed at 7, 14, and 28 days after completion of distraction, respectively. Immediately after the animals were killed, the right mandibles were harvested en block. Immunohistochemical staining was processed for observation of the VEGF expression, and double immunofluorescent staining was also processed for detection of the co-expression of osteocalcin and VEGF's two distinct receptors (VEGFR-1 and VEGFR-2). RESULTS: At 7 and 14 days after distraction, the expressions of VEGF were significantly increased in the osteogenic cells of the distracted bone. Up to 28 days after distraction, VEGF was still expressed moderate in the osteoblastic cells of distracted bone. The co-expressions of osteocalcin / VEGFR-1 and osteocalcin / VEGFR-2 were observed in the distracted bone at 7 and 14 days after distraction. In the double immunofluorescent staining, the co-expression's level of osteocalcin / VEGFR-1 was more than that of osteocalcin / VEGFR-2. Conclusion: Taken together, this study suggested that VEGF plays an important role in the osteogenesis, and these osteoblastic cell-derived VEGF might act as autocrine growth factor during distraction osteogenesis. In the other word, the cellular components, such as osteoblasts and immature fibroblast-like cellsor mesenchymal cells in the distracted bone, might have autocrine growth activity during distraction osteogenesis.

Expression of vascular endothelial growth factor and its receptors in the distracted periosteum after mandibular distraction osteogenesis

Changes of urotensin Ⅱ in rat with chronic obstructive pulmonary disease / 基础医学与临床

Objective To investigate the roles of urotensin Ⅱ(U-Ⅱ) in rat with chronic obstructive pulmonary disease(COPD).Methods Male SD rats were randomly divided into chronic bronchitis group(group B),COPD group(group C),exsmoker group(group D) and control group(group A). The concentrations of U-Ⅱ in plasma,BALF and lung tissues were measured by RIA.Lung function and pathological analysis of lung specimens were performed.Results There was no significant difference of plasma U-Ⅱ levels among groups.The BALF levels of U-Ⅱ was 1.7,2.2,4.7,4.9 folds higher than plasma levels in group A,B,C and D respectively(all(P