ABSTRACT
Objective:To investigate the effect of long non-coding RNA 068 (lncRNA 068) on the migration of a melanoma cell line A375, and to explore its mechanism of action.Methods:From December 2015 to November 2020, 21 patients with pathologically confirmed cutaneous melanoma were collected from Department of Dermatology, Affiliated Hospital of Nantong University, and quantitative PCR (qPCR) was performed to determine the expression of lncRNA 068 in melanoma and paracancerous tissues. LncRNA 068 was overexpressed or knocked down via lentiviral transfection in A375 human melanoma cells in the following experiments. Specifically, A375 cells were divided into lentiviral vector (LV) -green fluorescent protein (GFP) group and LV-lncRNA 068 group to be transfected with a GFP-expressing LV and a LV containing lncRNA 068 respectively in the overexpression experiment, and were divided into LV-LacZ short hairpin RNA (shRNA) group and LV-lncRNA 068 shRNA group to be transfected with a LV containing the reporter gene LacZ-specific shRNA and a LV containing the lncRNA 068-targeting shRNA respectively in the low-expression experiment, with the LV-GFP group and LV-LacZ shRNA group serving as the control groups. Transwell and scratch assays were performed to evaluate cell migration, EdU cell proliferation assay and cell counting kit-8 (CCK8) assay to determine the proportion of proliferative cells and cell viability respectively, and immunofluorescence staining was conducted to evaluate epithelial-mesenchymal transformation in the above groups. Lentivirus-transfected A375 cells from the above groups were inoculated into the axillae of BALB/c nude mice, and tumor volume was measured and calculated every 3 days. After 30 days, all mice were sacrificed, and tumor tissues were resected to measure the tumor volume and weight. Cultured B16F10 cells were subcutaneously inoculated into the back and foot of BALB/c nude mice to construct mouse models of subcutaneously transplanted B16F10 melanoma. After 2 weeks, the mice were sacrificed, and qPCR and Western blot analysis were performed to determine the mRNA expression of inflammatory factors in transplanted B16F10 melanoma and paracancerous tissues, and expression of IκB kinase (IKK) /P65 signaling pathway-related proteins, respectively. Comparisons between 2 groups were done by t test, and comparisons of tumor volume and weight at different time points among groups were done by repeated measures analysis of variance. Results:qPCR showed that the relative expression of lncRNA 068 was significantly lower in human melanoma tissues and transplanted B16F10 murine melanoma tissues (0.414 ± 0.109, 0.717 ± 0.041, respectively) than in the corresponding paracancerous tissues (1.050 ± 0.103, 1.011 ± 0.023, t = 19.48, 10.83, respectively, both P < 0.001). Transwell and scratch assays both showed that the cellular migratory ability was significantly lower in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.01), and significantly higher in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Immunofluorescence assay showed significantly increased fluorescence intensity of E-cadherin and decreased fluorescence intensity of N-cadherin in the LV-lncRNA 068 group compared with the LV-GFP group (both P < 0.001), but significantly decreased fluorescence intensity of E-cadherin and increased fluorescence intensity of N-cadherin in the LV-lncRNA 068 shRNA group compared with the LV-LacZ shRNA group (both P < 0.05). In vivo tumor formation experiment in nude mice showed that there were no significant differences in the volume or weight of melanoma between the LV-lncRNA 068 group and LV-GFP group (both P > 0.05), as well as between the LV-lncRNA 068 shRNA group and LV-LacZ shRNA group (both P > 0.05). As qPCR and Western blot analysis showed, the mRNA and protein expression of interleukin-10 (IL-10) and claudin-1 in A375 cells were significantly higher in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.05), but significantly lower in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Compared with the paracancerous tissues, B16F10 melanoma tissues showed significantly decreased mRNA expression of IL-10 ( P < 0.01), but significantly increased mRNA expression of IL-6 and tumor necrosis factor-α, as well as protein expression of phosphorylated P65 and phosphorylated IKK ( P < 0.01) . Conclusion:Overexpression of lncRNA 068 can inhibit the migration of A375 melanoma cells, and may affect the development of inflammation and inhibit the epithelial-mesenchymal transformation by inhibiting the IKK/P65 signaling pathway.
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Objective:To explore the influence of early essential newborn care (EENC) on breastfeeding after vaginal delivery.Methods:Women with full-term vaginal delivery at Jiaxing Women and Children's Hospital Affiliated to Wenzhou Medical University from January to June 2019 were enrolled in this prospective randomized controlled trial. These participants and their neonates were assigned randomly to receive early skin-to-skin contact for 90 min after delayed umbilical clamping (EENC group) or routine nursing care after delivery and skin-to-skin contact for 30 min (control group). The main outcomes were successful initiation and duration of first-time breastfeeding, initiation time of second-stage lactation, self-efficacy score of breastfeeding, acceptance and satisfaction score of breastfeeding, and the rate of exclusive breastfeeding at 24, 48, 72 h, and 42 d postpartum. The secondary outcomes included the onset time of first deep sleep of the newborn, the baby's first breast-fed stool, and the neonatal weight within 4 days of birth. Two independent samples t-test and Chi-square test were used for statistical analysis. Results:A total of 200 women with full-term vaginal delivery were recruited and 117 (61 in EENC group and 56 in control group) completed the study. (1)Main outcomes: Compared with the control group, the EENC group had earlier initiation of the first breastfeeding [(25.5±6.9) vs (33.2±7.2) min, t=5.902] and the second stage of lactation [(72.1±3.3) vs (78.6±4.3) h, t=9.504], longer duration of the first breastfeeding [(40.6±5.2) vs (32.6±6.1) min, t=7.806], and higher self-efficacy score [(122.5±2.1) vs (98.2±3.1) scores, t=50.660], acceptance score [(21.8±3.1) vs (15.3±4.3) scores, t=9.444], satisfaction score of breastfeeding [(23.2±2.1) vs (18.8±3.1) scores, t=9.278], and higher rate of exclusive breastfeeding at 24, 48 and 72 h after delivery[43% (26/61) vs 21% (12/56), 59%(36/61) vs 36%(20/56), 89%(54/61) vs 64%(36/56); χ 2 were 5.980, 6.353, and 9.663, respectively], all P<0.05. (2)Secondary outcomes: In the EENC group, the onset time of first deep sleep [(90.1±10.6) vs (118.3±9.9) min, t=7.645] and the the baby's first breast-fed stool [(57.1±6.4) vs (66.4±5.5) h, t=8.435] were earlier; neonatal weight at 3 d [(3 720±329) vs (3 558±412) g, t=2.382] and 4 d after birth [(3 778±289) vs (3 562±315) g, t=3.857] were significantly heavier, comparing with the control group (all P<0.01). Conclusions:EENC can improve multiple short-term breastfeeding indicators and maternal acceptance and satisfaction of breastfeeding in women after full-term vaginal delivery.
ABSTRACT
Objective To explore the effects of early essential newborn care (EENC) on short-term maternal and neonatal health and to evaluate the satisfaction and acceptability of both patients and medical staff in the operating room regarding the implementation of EENC during term cesarean section(CS). Methods Two hundred gravidas who underwent CS in Wenzhou Medical University Affiliated Women and Children Hospital from January 2018 to April 2018 were recruited in this prospective study. According to the odd or even number of their medical records, these women were randomly divided into EENC or control group, 100 in each. EENC was offered to those in the EENC group immediately after birth, including drying the newborn immediately and thoroughly, mother–infant skin-to-skin contact at least 90 min and initiating the first breastfeeding, and delayed cord clamping until l-3 min after birth. Routine neonatal care was provided to the control group, including regular drying, insufficient skin-to-skin contact and cord clamping within 1 min after birth. Differences were compared between the two groups in the incidence of abnormal pulse, low oxygen saturation, hypothermia and mild asphyxia from the neonatal aspect and the incidence of postpartum hemorrhage and early initiation of breastfeeding from the maternal aspect. The acceptability and satisfaction of all the participants and the medical staff were also analyzed. t-test and Chi-square test were used as statistical methods. Results All of the 200 participants were finally analyzed. In the EENC group, the incidence of neonatal hypothermia [2% (2/100) vs 13% (13/100), χ2=8.721, P=0.003] and maternal postpartum hemorrhage [1% (1/100) vs 6% (6/100), χ2=5.701, P=0.035] were lower comparing to the control group, while the initiation rate of early breastfeeding was higher [56% (56/100) vs 5% (5/100), χ2=61.352, P<0.001]. There was no significant difference in the incidence of neonatal abnormal pulse, low oxygen saturation or mild asphyxia between the two groups (all P>0.05). The scores of maternal satisfaction (24.6±0.4 vs 23.4±1.9, t=6.443, P=0.001) and acceptability (24.3±0.5 vs 23.5±1.4, t=5.436, P=0.001) in the EENC group were also significantly higher than those in the control. For the obstetric operation team, the acceptability scores among obstetricians (22.6±0.8 vs 21.6±1.3, t=2.379, P=0.019), instrument nurses (23.2±0.9 vs 21.3±1.1, t=13.592, P<0.001) and anesthetists (22.6±0.9 vs 21.5±1.7, t=5.625, P=0.001) in the EENC group were higher than those in the control group. However, no significant difference was observed in the acceptability among rounding nurses between the two groups (P=0.086). Conclusions EENC during CS could stabilize the neonatal temperature, reduce postpartum hemorrhage, promote early breastfeeding and improve the maternal satisfaction as well as the acceptability of both obstetric operation team and puerperae. There is a clinical significance to promote EENC during CS.
ABSTRACT
Objective To evaluate the effects of kangaroo mother care (KMC) during cesarean section on neonatal health parameters in the "golden hour" of late preterm infants. Methods A total of 120 singleton pregnant women undergoing preterm cesarean section (34-36+6 gestational weeks) from January 1 to December 31, 2018 in Jiaxing Women and Children's Hospital Affiliated to Wenzhou Medical University were prospectively enrolled and divided into two groups (observation and control groups) with 60 cases in each by random number table method. During cesarean section, the both groups received the thorough drying immediately and delayed cord clamping, in addition, the observation group adopted KMC, while the control group received routine maternal and infant care. Several parameters including changes in body temperature, crying, respiration, oxygen saturation and other vital signs, as well as the incidence of hypothermia, transfer to the Department of Neonatology and the success rate of early breastfeeding initiation during the "golden hour" were compared between the two groups. Satisfaction of obstetricians, pediatricians, the pregnant women and their families to KMC and routine maternal and infant care were compared. Two-independent sample t test and Chi-square test were used as statistical methods. Results In total, 113 cases (58 in the observation group and 55 in the control group) were finally analyzed. The body temperature of the late preterm infants at 5 and 10 min after birth in the observation group was higher than that in the control group (36.5 vs 36.0 ℃, 36.4 vs 35.8 ℃, t=11.756 and 7.512, both P<0.05), and the body temperature during 15 min-1 h after birth was also higher in the observation group but without statistical significance (all P>0.05). The observation group had a lower incidence of hypothermia [0% (0/58) vs 15% (8/55), χ2=9.079, P<0.001] and a higher success rate of early breastfeeding initiation [86% (50/58) vs 58% (32/55), χ2=11.137, P<0.001] than the control group. The satisfaction scores of obstetricians (19.3±1.1 vs 13.4±1.9, t=20.517), pediatricians (18.2±1.8 vs 12.6±2.1, t=15.382), gravidas (19.6±0.4 vs 13.6±1.6, t=27.723) and their family members (18.2±0.9 vs 15.3±1.1, t=11.535) were all higher in the observation group than those in the control group (all P<0.001). Conclusions KMC during cesarean section can improve the body temperature of late premature infants in the "golden hour", reduce the occurrence of hypothermia, facilitate early initiation of breastfeeding and improve the satisfaction of doctors and patients on health care.