Effects of RCCS simulated microgravity on proliferation and cell cytoskeleton of human HaCaT keratinocyte / 中华急诊医学杂志

Objective To investigate the effects of simulated microgravity by RCCS on proliferation and cell cytoskeleton of human HaCaT keratinocyte. Methods The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and human HaCaT keratinocytes were divided randomly(random number) into the simulated microgravity group (SMG) and normal gravity group (NG). HaCaT cells in the two groups were harvested respectively after 32, 36 and 42 h culture. The HaCaT cells proliferation and cycles were detected by flow cytometry, the concentration of hb-EGF in supernatant was detected by ELISA, and the cell cytoskeleton was observed after 42 hours' culture under laser confocal microscope with FITC-labeled technique. SPSS 23.0 statistical software was used for statistical analysis, and P <0.05 was considered statistically significant. Results The flow cytometry showed that the proportions of human HaCaT keratinocytes in G1 and G2/M phases were increased while the proportion of HaCaT cells in S stage was decreased significantly after 32, 36 and 42 h RCCSculture compared with those in the normal gravity group. The HaCaT cells in G1 stage were declined along with incubation time. ELISA results showed that the hb-EGF concentration in HaCaT supernatant under simulated microgravity culture for 24 and 36 h was lower than that in the normal control group (P<0.01). The laser confocal microscope revealed that the HaCaT fluorescence intensity was decreased,and there were disordered microfilaments, structural ambiguity, pseudopodia reduction and irregularshape among FITC-labeled HaCaT cells cultured 42 h in RSSC compared with the normal gravity group.Conclusions RCCS simulated microgravity environment could inhibit the cell cycle transformation and proliferation of human HaCaT keratinocyte, affect the keratinocyte-secreting function, and induce alterations of the cell cytoskeleton.

Study of HB EGF,C-erbB-2 test in differential diagnosis of ovarian benign and malignant tumor and determine the meaning of a complex condition / 国际检验医学杂志

Objective To study HB‐EGF ,C‐erbB‐2 test in differential diagnosis of ovarian benign and malignant tumor and de‐termine the meaning of a complex condition .Methods Totally 80 cases of patients with ovarian tumors were collected from August 2014 to July 2015 ,including 20 cases benign ovarian tumor ,and 40 cases epithelial ovarian cancer ,13 cases of Ⅰ period ,Ⅱ period in 14 cases ,Ⅲ ~ Ⅳ period 13 cases .At the same time ,normal endometrium ovarian tissue from 30 cases volunteers were collected .HB EGF ,C‐erbB‐2 expression in ovarian tissue were detected by using flow cytometry .The immune histochemical method was used to detect C‐erbB‐2 expression from 40 examples carcinoma ,30 cases of normal endometrium intrauterine membrane .Results The rate of positive cells of HB‐EGF ,C‐erbB‐2 was significantly higher than that of benign ovarian tumor(P< 0 .05) ;C‐erbB‐2 for lower ex‐pression in normal endometrium ,endometrial carcinoma in content was higher than the normal uterus express ,the difference was statistically significant(P< 0 .05) ;Endometrial cancer of different pathological staging ,C‐erbB‐2 expression was different also ,in‐cluding C‐erbB‐2 expression in endometrial carcinoma of stage Ⅰ ,Ⅱ significantly below Ⅲ ,Ⅳ period ,the difference was statistical‐ly significant(P< 0 .05) .Conclusion The high expression of C‐erbB‐2 protein is associated with the occurrence and development of endometrial carcinoma ,C‐erbB‐2 testing could help to clinically evaluate the biological behavior and prognosis of endometrial carci‐noma .

Expression and its correlations of caudal type homeobox transcription factor-2, heparin-binding epidermal growth factor-like growth factor and bone morphogenetic protein-4 in Barrett's esophagus / 中华消化杂志

Objective To explore the function and significance of caudal type homeobox transcription factor-2 (CDX)-2,heparin-binding epidermal growth factor-like growth factor (HB-EGF) and bone morphogenetic protein 4 (BMP-4) of esophageal stromal tissues in the pathogenesis of Barrett's esophagus (BE).Methods A total of 116 patients were divided into groups according to gastroscopic finds and hematoxylin-eosin (H &E) staining of biopsy samples.They were divided into control group (n=29),RE group (n=32),BE group (n=35),RE treatment group (n=10) and BE treatment group (n=10).The expression of CDX 2,HB-EGF and BMP-4 in different esophageal mucosal lesions was detected by immunohistochemical staining and the changes of positive expression levels of CDX-2,HBEGF and BMP 4 were compared among groups.Variance analysis and chi-square analysis were performed to analyze the correlation among the three factors.Results The CDX-2 positive cell number of control group ((0.0±0.0)/high power field(HPF)),RE group ((43.1±10.6)/HPF),and BE group ((67.8±11.3)/HPF) increased in turn,and the differences among three groups were statistically significant (F=67.664,P＜0.01).The HBEGF positive ((6.4±1.4)/HPF,(39.4±13.5)/HPF,(55.8±13.9)/HPF) and BMP 4 positive ((0.0±0.0)/HPF,(22.6±6.4)/HPF and ((25.1± 10.3)/HPF) cell number of three groups had the same trend and the differences among three groups were statistically significant (F HB-EGF =22.925,FBMP-4 =10.463,both P＜0.01).Except the expression of BMP-4 between RE group and BE group,there were significant differences between every other two groups (LSD test,all P＜ 0.01).The expression of CDX 2,HB EGF,BMP-4 in RE treatment group ((21.7±1.7)/HPF,(16.6±5.0)/HPF and (9.2±1.0)/HPF) and BE treatment group ((51.4±8.7)/HPF,(31.0± 10.4)/HPF and (12.7±3.9)/HPF) were lower than those in RE group and BE group respectively,the differences were also statistically significant (LSD test,all P＜0.05).In RE group,the positive rate of CDX-2 (31.2％ (10/32)) was significantly lower than that of HBEGF and BMP4 (62.5％ (20/32) and 56.2％(18/32)),and the differences were statistically significant (x2=6.275 and 4.063,both P＜0.05).However in BE group,there was no statistically significant difference in the positive rate among CDX-2(85.7％(30/35)),HB-EGF (88.6％(31/35)) and BMP-4 (74.3％(26/35),all P＞0.05).Conclusions CDX-2,HB-EGF and BMP-4 may play an important role in the pathogenesis of RE and BE.HB-EGF and BMP-4 may be involved in the early episodes of BE genesis and have promotion effects on CDX-2 expression.HB-EGF and BMP-4 may be the new target in the research and treatment of BE.

Heparin-binding epidermal growth factor-like growth factor and necrotizing enterocolitis / 国际儿科学杂志

Necrotizing enterocolitis(NEC) is a common and devastating gastrointestinal disease that occurs predominantly in premature infants.Despite various advances in management,the mortality of this disease remains high.During the last decade,studies have shown that heparin-binding epidermal growth factor-like growth factor (HB-EGF),can protect intestinal epithelial cells from various forms of injury in vitro.Furthermore,studies using both an intestinal ischemia-reperfusion injury model in adult rats and neonatal rat model of NEC,have shown that HB-EGF can protect the intestines from injury.Present studies provide a new research direction in the prevention and treatment.

The Effect of Antiproliferative Factor on the Proliferation of Bladder Cancer Cell and the Expression of c-Jun and HB-EGF / 中国医科大学学报

Objective To clarify the effect of antiproliferative factor（APF）on the proliferation of bladder cancer cell line T-24.Methods Active or no active APF with different concentrations was added to cultured bladder cancer T-24 cells.Methyl thiazolyl tetrazolium（MTT） assay was used to determine the proliferation of T-24 cells.The expression of c-Jun and HB-EGF were examined by Western blot.Results MTT showed that there was significant difference in the proliferation with the concentration of AFP increased from 0.5 μg /ml to 1.0 μg /ml.Compared with the negative control group,the absorption value of no active APF group showed no change（P 〉 0.05）.Western blot indicated that the c-Jun and HB-EGF expression were down-regulated with the concentration of active APF increased（P 〈 0.05）.No difference in their expression was found in no active APF groups.Conclusion Active APF may decrease the expression of c-Jun and HB-EGF,so as to inhibit the proliferation of bladder cancer cells.Our experiment might provide evidence for the therapeutic applications of APF for the bladder cancer.

Epidermal Hyperplasia and Elevated HB-EGF are More Prominent in Retinoid Dermatitis Compared with Irritant Contact Dermatitis Induced by Benzalkonium Chloride

BACKGROUND: 'Retinoid dermatitis' is a retinoid-induced irritant contact dermatitis (ICD). The mechanism of retinoid dermatitis may be different from that of other ICDs. However, it remains uncertain how topical retinoid induce ICD. OBJECTIVE: We compared several aspects of contact dermatitis induced by topical retinol and benzalkonium chloride (BKC) on hairless mice skin. METHODS: 2% retinol or 2.5% BKC was applied to hairless mice and transepidermal water loss (TEWL), ear thickness, histologic and immunohistochemical findings were compared. We also compared mRNA expression of inflammatory cytokines, epidermal differential markers, cyclooxygenases (COXs) and heparin binding epidermal growth factor like growth factor (HB-EGF). RESULTS: Topical application of 2% retinol and 2.5% BKC increased TEWL and ear thickness in similar intensity. Epidermal hyperplasia was more prominent in retinol treated skin. Proliferating cell nuclear antigen, involucrin and loricrin expression were higher in retinol-treated skin than in BKC-treated skin. Filaggrin, however, was more expressed in BKC-treated skin. The mRNA expression of IL-8, TNF-alpha, COX-2, involucrin, loricrin and filaggrin were increased in both retinol- and BKC-treated skin in similar intensity. HB-EGF was more significantly increased in retinol-treated skin. CONCLUSION: Elevated HB-EGF and epidermal hyperplasia are more prominent features of retinoid dermatitis than in BKC-induced ICD.

Effect of exogenous gonadotropin on HB-EGF expression in endometrium / 대한산부인과학회지

OBJECTIVE: Gonadotropin hyperstimulation for in vitro fertilization and embryo transfer (IVF-ET) has been considered to be harmful to the uterine receptivity. The role of HB-EGF has been implicated in the mouse embryonic implantation. In the present study, we assessed the effect of gonadotropin hyperstimulation on the expression of HB-EGF mRNA in murine endometrium. METHODS: Competitive, reverse transcription-PCR was performed to evaluate the expression of HB-EGF mRNA in the hyperstimulated murine endometrum from pregnant uteri of gestation day (g.d.) 5.5 after the injection of saline (placebo), pregnant mare serum gonadotropin (PMSG) 5 IU and 10 IU. RESULTS: HB-EGF expression was observed at 10-4-10-6 dilution concentrations of internal standard, and dual bands (423/360 bp) were observed at 10-5 dilution concentration. The expression of HB-EGF mRNA showed no significant change in the endometrial expression in PMSG groups, compared to the control group, on g.d. 5.5. There was no dose-response relationship between the two PMSG groups. CONCLUSION: Our data reveals that ovarian hyperstimulation using gonadotropin does not influence the expression of HB-EGF in the murine endometrium. Further studies are necessary to elucidate which stage of embryonic implantation and the expression of other substance is affected by the ovarian hyperstimulation.

The expression of HB-EGF (heparin binding epidermal growth factor) and TGF-A (transforming growth factor alpha) during liver regeneration after partial hepatectomy in mice

The expression of HB-EGF (heparine binding epidermal growth factor) in cells of mice increased just after partial level removal in hapatectomy and reached maximal value 3 hours later. TGF-A expression in mice cells after 2/3 hepatectomy reduced in comparing with control in 3 first hours later and increased in 6 hours later

Effects of Heparin-Binding Epidermal Growth Factor on the Expression of MMP-9 and ATPase gamma-subunit mRNA in the Mouse Embryo / 대한불임학회지

OBJECTIVES: The purpose of this study was to evaluate effects of heparin-binding epidermal growth factor (HB-EGF) on the rate of blastocyst formation and hatching in the mouse embryos and the expression of matrix metalloproteinase-9 (MMP-9) and ATPase g-subunit mRNA. METHODS: Late 2-cell mouse embryos was cultured for 72 hours in HTF medium containing with 1, 10, and 100 ng/ml HB-EGF. The mRNA expression level of MMP-9 and ATPase gamma-subunit was detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The rate of hatching was significantly higher (p<0.05) in group containing with 1 ng/ml HB-EGF than other groups. Also, the rate of hatched blastocyst was significantly higher (p<0.05) in 10 ng/ml. The mRNA expression level of MMP-9 mRNA was not shown any difference among groups, but ATPase g-subunit was higher than other groups. CONCLUSIONS: Taken together these results suggest that HB-EGF has the positive effect to promote the blastocyst formation and hatching process and influences the blastocoel expansion by promoting the ATPase mRNA expression in the mouse embryos.

Regulation of tazarotene on HB-EGF in hyperproliferative epidermis of psoriasis / 西安交通大学学报(医学版)

Objective To investigate the mechanism of tazarotene in active psoriasis vulgaris. Methods HB-EGF mRNA in active psoriatic lesions before and 10 days after the treatment with tazarotene was detected by hybridization in situ. Results There was nearly no expression of HB-EGF mRNA in psoriatic lesions (9.1%); after the treatment with tazarotene, there was expression of HB-EGF not only in basal layer (95.5%), but also focal expression in suprabasal layers of epidermis (77.3%). Conclusion Tazarotene can inhibit proliferation and induce apoptosis of keratinocytes though upregulating expression of HB-EGF in psoriatic epidermis.