Objective To observe the permeability and stability of the transfection of antisense oligonucleotide (ASODN) hybridized epidermal growth factor receptor (EGFR) to retinal glial cells (RG).Methods Phosphorothioate and unmodified EGFR ASODN conjugated with 5′-isothioc yanate (5′-FITC) were encapsulated with or without lipofectin, and then added into human retinal glial cells culture media. The cellular permeability and stability of the transfection were observed by fluorescence microscopy in fixed cells.Results In the absence of lipofectin, phosphorothioate and unmodified EGFR ASODN were found in a few RG cells at 30 minutes, and in about 50% RG cells at 4 hours. Phosphorothioate EGFR ASODN were kept in RG cells for 3-4 hours and disappeared at about 8 hours. In the presence of lipofectin, phosphoro thioate and unmodified EGFR ASODN were found in a few RG cells at 15 minutes and about 70%-80% RG cells at 4 hours. Phosphorothioate EGFR ASODN were kept in cells for 10-12 hours, and phosphorothioate and unmodified EGFR ASODN were disapp eared at about 14 hours and 4 hours respectively.Conclusion 5′-FITC EGFR ASODN encapsulated with lipofectin could enter RG cells and express stably in RG cells. (Chin J Ocul Fundus Dis，2003，19：52-54)
OBJECTIVE: To investigate the efficiency of recombinant human epidermal growth factor (rhEGF) on burn wound healing and to explore the effective density of the ointments. METHODS: A total of 120 cases of burn in superficial II degree and profound II degree were randomly divided into 2 groups. In the first group of 15 cases of superficial II degree, the wounds were treated by rhEGF ointments of different density, 0.5 microgram/g, 10 micrograms/g and 50 micrograms/g, to screen out the effective density. And in the other 105 cases of the second group, optimal density of the ointments based on the result of the first group were employed to treat the burn wound in superficial II degree and profound II degree, with the self-corresponding wounds of the same degree as control, to study the efficiency of rhEGF on wound healing, according to the wound healing time, and adverse reaction of the ointment. RESULTS: In the first group, the average healing time of superficial II wound treated by ointments of 10 micrograms/g and 50 micrograms/g significantly shortened when compared with that treated by ointments of 0.5 microgram/g(P lt; 0.01), but there was no obvious difference between the cases treated by ointments of 10 micrograms/g and 50 micrograms/g. In the second group, the healing time of superficial II wound treated by ointments of 10 micrograms/g was (8.39 +/- 2.25) days, (9.52 +/- 2.56) days in the control (P lt; 0.01); and healing time of profound II burn treated by ointments of 10 micrograms/g was (16.80 +/- 2.99) days, (18.27 +/- 3.17) days in the control (P lt; 0.01). And healing rates of burn wound at different periods were higher than those of the control. CONCLUSION: The above results indicate that rhEGF ointments can enhance burn wound healing significantly, and the ointment of 10 micrograms/g is a good choice for clinical application.
To investigate the significance of epidermal growth factor receptor (EGFR) in thyroid carcinoma, the expression of EGFR in 81 samples of thyroid carcinoma were determined by immunohistochemical SP method and comparison among thyroid carcinoma, thyroid adenomas and normal thyroid tissue adjacent to the cancer were made. The results showed: EGFR expression was positive in 45 cases (55.6%) of thyroid carcinoma with no positive expression either in thyroid adenomas or normal thyroid tissue adjacent to the cancer (Ｐ＜0.01). There were no statistical differences between EGFR positive rate and thyroid carcinomatous pathological type, clinical stage, depth of invasion, lymph node metastasis or patients′ postoperative survival time (Ｐ＞0.05). This data suggests that expression of EGFR in thyroid carcinoma is associated with its autonomous growth and malignant phenotype, but it is probably not a useful index for assessing the biological behavious and prognosis of thyroid carcinoma.
ObjectiveTo evaluate the effect of heparin binding epidermal growth factor-like growth factor (HB-EGF) on liver regeneration after partial orthotopic liver transplantation. MethodsFourty SD rats were used to establish the model of partial orthotopic liver transplantation with ameliorated two-cuff technique. Then all the rats were divided into 2 groups: experiment group and control group. Twenty rats of experiment group were administered 500 μg/kg HBEGF via vena caudalis immediately after operation twice a day, while the same volume of saline was administered to the rats in control group. Five rats in each group were selected randomly and killed at the 6th hour, day 2, 4 and 7 after operation, respectively. The serum levels of albumin (Alb) and alanine aminotransferase (ALT) in the blood sample were detected. Every liver was removed and weighed. The expression of Ki67 was detected by using immunohistochemistry assay. The regeneration activity of hepatocytes was evaluated by flow cytometry. ResultsThe wet weights of liver in experiment group were all significantly higher than that in control group at the 6th hour, day 2 and 4 after transplantation (P＜0.05). The serum levels of ALT were significantly lower in experiment group than those in control group at the 6th hour, day 2， 4, 7 after operation (P＜0.05), while the levels of Alb were significantly higher on day 4 and 7. The proliferating index and Ki-67 labeling index of graft in experiment group were higher than those in control group on day 2 and 4 after transplantation （2 d: P＜0.01; 4 d: P＜0.05）. ConclusionHBEGF could promote the regeneration of rat hepatocytes after partial liver transplantation.
Objective To investigate the activation and role of signal transduction pathway of epidermal growth factor (EGF)-epidermal growth factor receptor (EGFR)-mitogen activated protein kinase (MAPK) in proliferation of human retinal pigment epithelial (RPE) cells. Methods Human RPE cells were stimulated with 0.1%，10% foetal calfserum (FCS) and EGF(0.1, 1, 10, 50 and 100 ng/ml)in 0.1% FCS Dulbeco′s modified Eagle′s medium (DMEM) and in 10% FCS DMEM for 3 days, respectively. Immunohistochemical staining and in situ hybridization were used to observe the expressions of EGFR protein and EGFR mRNA，respectively. Activation of MAPK was detected by immunohistochemical method with specific anti-phosphorylated ERK 1/2 antibody. Results The optimal concentrations of EGF were 10 ng/ml in 0.1% FCS DMEM and 1 ng/ml in 10% FCS DMEM. After 3 days of stimulation with EGF, phosphorylated ERK 1/2 staining was detectable in nucleus of RPE cells, whereas cells presented immunostaining for phosphorylated ERK 1/2 in the cytoplasm before stimulation. Conclusions EGF may improve the expression of EGFR protein and EGFR mRNA of RPE cells, and induced MAPK nuclear translocation in a concentration-dependent manner. EGF-EGFR-MAPK signal transduction pathway may play a key role in RPE cells proliferation, and serum exerts an important acceclerating function in the process. (Chin J Ocul Fundus Dis，2004，20：67-132)
Objective:To study the effects of growth factor on the proliferation of the cultured huamn retinal glial cells. Methods:EGF(0.5～100.0ng/ml) and NGF (0.5～10.0ng/ml) were added to cultures of human retinal glial cells and the proliferation rates of the cells were measured by MTT method. Results:EGF at a dosage ranging from 0.5ng/ml to 100.0ng/ml and NGF (0.05～10.0ng/ml) stimulated the cellular proliferation effectively with their EC 50 of 17ng/ml and 0.7 ng/ml respectively. Conclusion:Both EGF and NGF NGF had an effective stimulation on human retinal glial cell proliferation.They may play a role in the formation of PVR. (Chin J Ocul Fundus Dis,1998,14:33-34)
OBJECTIVE: To investigate the mechanism of overhealing alleviation by salvia miltiorrhiza (SM) in wound healing. METHODS: Fibroblasts were cultured in vitro, and SM was applied with different concentrations (40, 80, 160 and 320 micrograms/ml) and time(the 1st, 2nd, 3rd, 4th and 5th days) to influence their autocrine. The levels of transforming growth factor-β1 (TGF-β1) and epidermal growth factor (EGF) were determined by ELIAS and radioimmunoassay respectively. RESULTS: The SM could inhibit autocrine of TGF-β1 by fibroblasts (P lt; 0.05). However, it did not affect autocrine of EGF (P gt; 0.05). CONCLUSION: The above results indicate that SM reduces overhealing by inhibiting the autocrine of TGF-β1 selectively.
Objective To investigate the differences in clinicopathological characteristics and prognostic survival of human epidermal growth factor receptor 2 (HER2) high expression, HER2 low expression and HER2 negative breast cancer. MethodWe retrospectively collected 1 560 female breast cancer patients who underwent surgical treatment at the Department of Breast and Thyroid Surgery in Renmin Hospital of Wuhan University between January 8, 2010 and December 31, 2015, and divided them into high expression group, low expression group and negative group according to HER2 expression, to compare the differences in clinicopathological characteristics among the three groups of breast cancer patients and to explore the factors influencing prognosis. Results The proportions of histological grade Ⅲ, tumor diameter >2 cm, lymph node metastasis, TNM stage Ⅲ, Ki67 high expression, and hormone receptor negative expression were higher in the high expression group than those in the low expression group and negative group (P<0.050); the proportions of histological grade Ⅲ, tumor diameter >2 cm, lymph node metastasis, and TNM stage Ⅲ were higher in the low expression group than those in the negative group (P<0.050). However, the proportions of Ki67 high expression and hormone receptor negative expression were lower than those of the negative group (P<0.050). The 5-year disease-free survival rate were 85.6%, 80.3% and 74.5% for the high expression, low expression and negative group, respectively, and the 5-year overall survival rate were 90.4%, 86.0% and 80.7%, respectively. The results of multivariate Cox proportional hazard model showed that patients with high histological grade, late TNM stage, Ki67 high expression and weaker HER2 expression intensity had worse 5-year disease-free survival (P<0.050); patients with older age, high histological grade, lymph node metastasis, late TNM stage, Ki67 high expression and weaker HER2 expression intensity had worse 5-year overall survival (P<0.050). Conclusions The intensity of HER2 expression affects the 5-year disease-free survival and overall survival of breast cancer patients, and the higher the intensity of HER2 expression, the better the 5-year disease-free survival and overall survival, while the weaker the HER2 expression, the worse the 5-year disease-free survival and overall survival.
In order to investigate the function of epidermal growth factor (EGF), the following experiments were performed. Thirty white rats were chosen and divided into 3 groups. In the back of each rat, two 2 cm x 2 cm wounds were made bilaterally, the skin and subcutaneous tissue was removed. EGF were used in one of the two wounds randomly, while those without EGF usage in control. After 1, 2 and 3 weeks, the rats were sacrificed. The area of the wounds was measured, and the healing time of each wound was recorded. The results showed that the healing time of EGF group was 14.6 days while that of control group was 18.5 days (P lt; 0.01). Furthermore, the DNA, protein and hydroxyproline contents of EGF group were higher than those of the control group (P lt; 0.01). It was suggested the EGF could accelerate wound healing and shorten the healing time.
Objective To explore the effect of membrane surface nucleolin （NCL） on activation of epidermal growth factor receptor （EGFR） signaling. Methods Immunohistochemistry was used to identify the expressions of membrane surface NCL or EGFR in pallilary thyroid carcinoma （PTC） tissues. The level of phosphorlated EGFR in TPC-1 cells was observed by Western blot. TPC-1 cells invasion capacity was detected by Transwell assay. Results The posi-tive expression rates of membrane surface NCL and EGFR in PTC tissues were 100% （56/56） and 80.4% （45/56） respe-ctively， while the expressions of NCL and EGFR were related with lymph node metastasis （P＜0.05）. There was posi-tive correlation between the expressions of NCL and EGFR （r=0.635， P＜0.01）. Western blot showed that anti-NCL or anti-EGFR of TPC-1 cells could inhibit the expression of phosphorlation EGFR （P＜0.01）. Transwell assay showed the number of membrane-invading cells were reduced significantly in anti-NCL group anti-EGFR group （P＜0.01）. Conclusions Membrane surface NCL may be a kind of indispensable component in activation of EGFR signaling， by which EGFR can participate in growth and invasion of tumors. NCL can be used as a target for developing a new field of tumor treatment.