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OBJECTIVES@#Silence of SET domain containing lysine methyltransferase 7 (SET7) alleviates myocardial tissue injury caused by ischemia-reperfusion. But the effects of SET7 on angiotensin II (Ang II)-induced myocardial fibroblast proliferation and the collagen synthesis are not clear. The purpose of this study was to explore the effect of SET7 on the proliferation and collagen synthesis of myocardial fibroblasts and its mechanisms.@*METHODS@#Myocardial fibroblasts were isolated and identified by immunofluorescence. Myocardial fibroblasts were randomly divided into 4 groups: a control group (cells were normally cultured), an Ang II group (cells were treated with 100 nmol/L Ang II for 24 h), a siCtrl group (cells were transfected with siRNA control and were then treated with 100 nmol/L Ang II for 24 h), and a siSET7 group (cells were transfected with siRNA SET7 and were then treated with 100 nmol/L Ang II for 24 h). Cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assay were used to evaluate cell proliferation. Real-time PCR was used to detect the mRNA levels of SET7, collagen I, collagen III, and α-smooth muscle actin (α-SMA). Western blotting was used to detect the protein expression of SET7, collagen I, collagen III, α-SMA, sonic hedgehog (Shh), ptched1 (Ptch1), and glioma-associated oncogene homolog 1 (Gli1).@*RESULTS@#Fluorescence microscopy showed positive vimentin staining, and myocardial fibroblasts were in good condition. As compared to the control group, the mRNA and protein levels of SET7 in the Ang II group were significantly upregulated; cell proliferation rate and EdU fluorescence intensity in the Ang II group were significantly increased; the mRNA and protein levels of collagen I, collagen III, and α-SMA were significantly upregulated (all @*CONCLUSIONS@#Silence of SET7 gene inhibits Ang II-induced proliferation and collagen synthesis of myocardial fibroblasts. Shh signaling pathway may be involved in this process.
Subject(s)
Angiotensin II/pharmacology , Cell Proliferation , Cells, Cultured , Collagen/genetics , Fibroblasts , Hedgehog ProteinsABSTRACT
Objective@#This study was designed to identify the relative factors, bacteriological profile and their antibiotic susceptibility pattern of neonatal sepsis.@*Methods@#A retrospective survey was conducted on the clinical information, pathogen identification and antibiotic sensitivity results of 425 newborns with neonatal sepsis admitted to Nanjing Maternity and Child Health Care Hospital from 2010 to 2017. Of the 425 positive blood-cultures, 148 (34.82%) were early-onset neonatal sepsis (EOS) and 277 (65.18%) were late-onset neonatal sepsis (LOS). Clinical information and pathogen identification were compared between EOS and LOS. Antibiotic sensitivity of gram negative organisms (G-) and gram positive organisms (G+) were also detected.@*Results@#The rates of premature delivery (78.70%, n=218), low birth weights (67.15%, n=186) and cesarean delivery (59.57%, n=165) were significantly increased in LOS (P<0.05) compared with those rates in EOS, which were 41.89% (n=62), 37.84% (n=56) and 46.62% (n=69). Parturients fever (18.24%, n=27) and meconium like amniotic fluid (25.68%, n=38) were significantly increased in EOS (P<0.05) compared with those rates in LOS, which were 7.94% (n=22) and 5.42% (n=15). Among the identified pathogen, the incidence of G- and G+ bacteria were 216 (50.83%) and 201 (47.29%) respectively, and the rest was Candida glabrata (1.88%, n=8). Escherichia coli 68 (16.00%) was the predominant isolate followed by Klebsiella pneumoniae (13.18%, n=56). The detection rate of Hemolytic staphylococcus (10.81%, n=16) was significantly increased in EOS (P<0.001) compared with LOS (1.44%, n=4). However, the incidence of Klebsiella pneumonia (5.88%, n=44) was higher in LOS (P=0.024) compared with EOS (8.11%, n=12). Most of the gram positive isolates exhibited high resistance to penicillin (90.32%-100.00%) and cephalosporin group antibiotics (25.00%-100.00%). Similarly, the majority of the gram negative isolates showed higher resistance to ampicillin (83.33%-100.00%), but susceptible to aminoglycosides (0-11.76%), quinolones (0-17.65%) and β-lactams (0-5.88%).@*Conclusion@#Among the study population, the percent of preterm, low birth weight and cesarean section were higher in LOS while parturients fever and meconium-like amniotic fluid were higher in EOS. The pathogens with the highest detection rate were Escherichia coli and Klebsiella pneumoniae. The results of antibiotic susceptibility test showed that common pathogens had high resistance to commonly used antibacterial drugs.
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Objective To investigate the effect of ellagic acid on human epidermal melanocyte melanogenesis and melanin transfer, and the mechanism thereof. Methods The human melanocytes and and keratinocytes were co-cultured and purified. After passing the second generation, cells of 1∶10 ratio were inoculated into the small dish (3 cm × 3 cm). The changes of melanin content and tyrosinase activity in melanocytes were detected before and after intervention with ellagic acid (100, 10 and 1 mg/L) for 48 h. The melanin transfer in cultured cells was detected by flow cytometry method. The 10 nmol/L arbutin was used as the positive control. Results The tyrosinase activity was down-regulated by ellagic acid in a dose-dependent manner. The ellagic acid can reduce the melanin content except for the 1 mg/L of ellagic acid. The melanin transfer was also inhibited by ellagic acid in a dose-dependence manner. Conclusion Ellagic acid can be used for skin-whitening cosmetic and the depigmenting effect might be due to the down-regulation of melanogenesis and melanin transfer.
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Objective To evaluate the efficacy and safety of 308-nm monochromatic excimer light for the treatment of amelanotic nevus.Methods Eighteen patients with amelanotic nevus were treated with 308-nm monochromatic excimer light twice a week for five consecutive weeks.Melanin content index (MCI) was measured in the lesions before and after the treatment.After the final exposure,a three-month follow-up was carried out.Results After the start of treatment,the MCI in lesions increased in a session-dependent manner and approximated to 100％ of that in perilesional normal skin after six sessions of treatment.The follow-up revealed a decrease trend in MCI after the end of the treatment,which was nearly equal to that in perilesional normal skin one month later,and dropped to about 90％ of that three months later.No side effects such as blisters or scar were observed during the treatment.Conclusions The 308-nm monochromatic excimer light is safe and effective for the treatment of amelanotic nevus.Re-exposure to 308-nm monochromatic excimer light after three months is recommended as consolidation therapy.
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Objective To investigate the ultrastructural characteristics of amelanotic melanocytes (AMMCs). Methods Individual hair follicles from normal human scalp were digested with collagenase type V, then washed in phosphate buffer saline. Hair-follicle cell suspensions were prepared by trypsin and cultured in a medium suitable for melanocyte growth. The keratinocytes were removed by differential trypsinization. Geneticin (100?g/mL) was used to eliminate contaminating fibroblasts. After 3 passages the cells were trypsinized, washed in phosphate buffer saline, and finally processed for transmission electron microscopy. Results Under transmission electron microscope, the cultured cells were round or oval-shaped with a single large nucleus and double-layered karyotheca. Abundant euchromosome but sparse heterochromosome was observed within the nucleus. There were various organelles in the cytoplasm, including mitochondria, rough endoplasmic reticulum (RER), ribosomes and abundant melanosomes of nearly uniform size. The electronic density granules distributed in a concentric pattern in most of the melanosomes. Colgi complexes were inconspicuous in the cells. Conclusions Compared to epidermal melanocytes, AMMCs from human hair follicles have different ultrastructural characteristics which implies their functional immaturity. AMMCs may serve as the depot for mature melanocytes.