Objective To investigate the impacts and mechanism of circular RNA polyribonucleotide nucleotidyltransferase 1(Circ-PNPT1)on glucose and lipid metabolism and offspring outcomes in rats with GDM through microRNA-345-3p(miR-345-3p)/lipid raft scaffold protein 2(FLOT2)axis.Methods 75 pregnant female rats were divided into normal control(NC)group,GDM group,si-NC group,si-Circ-PNPT1 group,and si-Circ-PNPT1+miR-345-3p-inhibitor group,with 15 rats in each group.Glucose and lipid metabolism indexes were detected in each group.FIns was detected by ELISA.The survival rate and body weight of fetal rats were compared in each group.The expression of Circ-PNPT1,miR-345-3p and FLOT2 mRNA in placenta was detected by qRT-PCR.The double luciferase reporter gene experiment verified the targeting relationship between miR-345-3p and Circ-PNPT1 and FLOT2.Western blot was used to detect the expression of FLOT2 protein in rat placenta.Results Compared with the NC group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expressions of Circ-PNPT1,FLOT2 in GDM group increased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p decreased(P<0.05).Compared with GDM group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expression of FLOT2 in si-Circ-PNPT1 group decreased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p increased(P<0.05);MiR-345-3p-inhibitor reversed the improvement of si-Circ-PNPT1 on GDM rats(P<0.05).Conclusions Knocking down Circ-PNPT1 can up-regulate miR-345-3p and down-regulate FLOT2 to improve glucose and lipid metabolism and offspring outcome in GDM rats.

Objective:To extract and evaluate the relevant evidence to improve the family discharge preparation of premature infants with bronchopulmonary dysplasia, so as to provide evidence-based basis for clinical formulation of scientific and effective discharge plans.Methods:All the evidence on the family discharge readiness of premature infants with bronchopulmonary dysplasia was collected from Chinese and English databases or websites, and the quality of various studies was evaluated. JBI′s evidence grading and recommendation level system (2014 edition) was used to extract and summarize the evidence.Results:A total of 21 articles were included, and 24 pieces of evidence were summarized, which were divided into seven themes: pre-discharge planning, environmental preparedness, parent education and training, support systems, feeding and nutrition, respiratory management, and discharge follow-up.Conclusions:For preterm infants with bronchopulmonary dysplasia, discharge readiness should be improved in terms of standardizing discharge criteria and implementing a discharge plan; improving family care preparation to ensure discharge support; strengthening health management and implementing a personalized plan; and continuing high-quality follow-up to ensure long-term health.

Objective To investigate the impacts and mechanism of circular RNA polyribonucleotide nucleotidyltransferase 1(Circ-PNPT1)on glucose and lipid metabolism and offspring outcomes in rats with GDM through microRNA-345-3p(miR-345-3p)/lipid raft scaffold protein 2(FLOT2)axis.Methods 75 pregnant female rats were divided into normal control(NC)group,GDM group,si-NC group,si-Circ-PNPT1 group,and si-Circ-PNPT1+miR-345-3p-inhibitor group,with 15 rats in each group.Glucose and lipid metabolism indexes were detected in each group.FIns was detected by ELISA.The survival rate and body weight of fetal rats were compared in each group.The expression of Circ-PNPT1,miR-345-3p and FLOT2 mRNA in placenta was detected by qRT-PCR.The double luciferase reporter gene experiment verified the targeting relationship between miR-345-3p and Circ-PNPT1 and FLOT2.Western blot was used to detect the expression of FLOT2 protein in rat placenta.Results Compared with the NC group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expressions of Circ-PNPT1,FLOT2 in GDM group increased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p decreased(P<0.05).Compared with GDM group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expression of FLOT2 in si-Circ-PNPT1 group decreased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p increased(P<0.05);MiR-345-3p-inhibitor reversed the improvement of si-Circ-PNPT1 on GDM rats(P<0.05).Conclusions Knocking down Circ-PNPT1 can up-regulate miR-345-3p and down-regulate FLOT2 to improve glucose and lipid metabolism and offspring outcome in GDM rats.

Objective:To extract and evaluate the relevant evidence to improve the family discharge preparation of premature infants with bronchopulmonary dysplasia, so as to provide evidence-based basis for clinical formulation of scientific and effective discharge plans.Methods:All the evidence on the family discharge readiness of premature infants with bronchopulmonary dysplasia was collected from Chinese and English databases or websites, and the quality of various studies was evaluated. JBI′s evidence grading and recommendation level system (2014 edition) was used to extract and summarize the evidence.Results:A total of 21 articles were included, and 24 pieces of evidence were summarized, which were divided into seven themes: pre-discharge planning, environmental preparedness, parent education and training, support systems, feeding and nutrition, respiratory management, and discharge follow-up.Conclusions:For preterm infants with bronchopulmonary dysplasia, discharge readiness should be improved in terms of standardizing discharge criteria and implementing a discharge plan; improving family care preparation to ensure discharge support; strengthening health management and implementing a personalized plan; and continuing high-quality follow-up to ensure long-term health.

Objective:To evaluate the effectiveness and safety of 3.5% lidocaine hydrochloride ophthalmic gel for eye surface anesthesia.Methods:A multicenter, randomized, double-blind, placebo controlled clinical trial was conducted in 10 hospitals in China from August 2018 to April 2019 under the approval of an Ethics Committee of Beijing Hospital (No.2018BJYYEC-014-02). A total of 220 fellow eyes of 220 subjects who received ocular surgery in one eye were actually enrolled according to a same included criteria in different institutes.The 220 eyes were randomly divided into control group and trail group.Two drops of lidocaine hydrochloride gel were dropped at about 5 mm away from corneal limbus at 6 o'clock direction of experimental eye of the trail group, and the blank gel was used in the eyes of the control group in the same way.The pain sensation was assessed with a 0.3 mm toothless micro forceps on conjunctiva within a specified time, and ''pain'' or ''no pain'' was answered by the subjects.The primary effective indexes, namely the number of eyes and percentage of ''no pain'' within 5 minutes following dropping, as well as the secondary indexes including the onset time point of the drug and the duration of anesthesia were recorded and evaluated.Safety evaluation took ocular and system adverse events into account.Results:Within 5 minutes after dropping, ''no pain'' occurred in 104 eyes (94.55%) and 29 eyes (26.36%) in the trail group and control group, respectively, showing a significant difference between the two groups ( χ2=106.948, P<0.001). And there was a significant difference in anesthesia onset time between the trail group and control group (40.0 seconds vs. 300.0 seconds) ( Z=-15.17, P<0.001). The duration of anesthesia was 860.5 (577.5, 1 180.0) seconds in the trail group and 676.0 (280.0, 1 401.0) seconds in the control group, with no statistically significant difference between the two groups ( Z=0.898, P>0.05). The incidence of adverse events in the trail group and control group were 5.45% (6/110) and 4.55% (5/110), respectively, without statistical significance between them ( P=1.000). Conclusions:The 3.5% lidocaine hydrochloride is a safe, effective, easy to use and high-quality surface anesthesia drug for eye surgery.

Objective: To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro. Methods: The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco′ s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 μmol/L capsaicin group, hypoxia+ 1.0 μmol/L capsaicin group, and hypoxia+ 10.0 μmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 μmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 μmol/L chloroquine, 10.0 μmol/L capsaicin, and 50 μmol/L chloroquine+ 10.0 μmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 μmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 μmol/L capsaicin group were treated with 10.0 μmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: (1) Compared with those of normoxia group, the protein expressions of LC3, Beclin-1, and TRPV1 were significantly increased in cardiomyocytes of hypoxia 3, 6, and 9 h groups (t_{3 h}=4.891, 5.890, 4.928; t_{6 h}=9.790, 6.750, 10.590; t_{9 h}=6.948, 6.764, 5.049, P<0.05 or P<0.01), which of hypoxia 6 h group were the highest (1.08±0.05, 1.12±0.10, 0.953±0.071, respectively). (2) The density of TRPV1 in cell membrane and inside the cardiomyocytes in hypoxia group was significantly increased with lump-like distribution, and the expression of TRPV1 was higher than that in normoxia group. (3) Compared with those of simple hypoxia group, the protein expression of Beclin-1 in cardiomyocytes of hypoxia+ 0.1 μmol/L capsaicin group was increased (t=10.488, P<0.01), while the protein expressions of LC3 and TRPV1 were increased without statistically significant differences (t=4.372, 3.026, P>0.05); the protein expressions of LC3, TRPV1, and Beclin-1 in cardiomyocytes of hypoxia+ 1.0 μmol/L capsaicin group and hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (t=15.505, 5.773, 13.430; 20.915, 8.054, 16.384; P<0.05 or P<0.01), which of hypoxia+ 10.0 μmol/L capsaicin group were the highest (2.33±0.09, 1.34±0.07, 1.246±0.053, respectively). (4) Compared with 0.585±0.045 in normoxia group, the cardiomyocyte viability in hypoxia group was significantly decreased (0.471±0.037, t=4.365, P<0.05). Compared with that in hypoxia group, the cardiomyocyte viability in hypoxia+ chloroquine group was further decreased (0.350±0.023, t=6.216, P<0.01), while 0.564±0.047 in hypoxia+ capsaicin group was significantly increased (t=3.489, P<0.05). Compared with that in hypoxia+ chloroquine group, the cardiomyocyte viability in hypoxia+ capsaicin+ chloroquine group did not significantly change (0.364±0.050, t=0.545, P>0.05). (5) Compared with 0.99±0.04 and 0.54±0.04 in simple hypoxia group, the protein expressions of LAMP-1 and LAMP-2 in hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (1.49±0.06, 0.81±0.05, t=12.550, 7.442, P<0.01). Conclusions TRPV1 can further promote the expression of autophagy-related proteins in hypoxic cardiomyocytes through autophagy-lysosomal pathway, enhance autophagy activity, and improve autophagic flow for alleviating early hypoxic cardiomyocyte injury.

Background Neovascular glaucoma (NVG) is a refractory disease characterized by neovascularization,and its pathogenesis mechanism is still unelucidated.Researches showed that muhicytokins and inflammatory factors are associated with neovascularization,however,the correlation of these factors with NVG worth attention.Objective This study was to detect the vescular endothelial growth factor (VEGF),transforming growth factor β1 (TGF-β1) and interleukin-6 (IL-6) levels in aqueous humor and plasma in NVG eyes and its significance.Methods A prospective cases-controlled study was performed under the approval of Ethic Committee of Shanghai East Hospital and informed consent of patients.Eight eyes of 8 NVG patients,10 eyes of 10 primary open angle glaucoma (POAG) patients and 10 eyes of age-related cataract (ARC) patients were entrolled in Shanghai East Hospital from May,2014 to March,2015.The periphery blood of 3-4 ml was collected from all the patients to prepare 0.3-0.4 ml serum,and 0.1-0.2 ml aqueous humor was collected during the surgery from each eye.The aqueous and serum levels of VEGF,TGF-β1 and IL-6 were measured by ELISA.Results The aqueous humor and plasma VEGF concentrations in NVG patients were (2 769.85 ± 390.88) pg/ml and (529.93 ± 95.20) pg/ml,respectively,which were higher than (208.12 ± 58.59) pg/ml and (219.28 ± 24.44) pg/ml in POAG patients as well as (158.88 ±12.35) pg/ml and (172.82±31.91)pg/ml in ARC patients,with significant differences among the three groups (aqueous:F=433.80,P＜0.01;plasma:F=103.84,P＜0.01).The levels of TGF-β1 in aqueous humor and plasma from NVG patients were (157.94±113.00)pg/ml and (3 895.78±2 318.00)pg/ml,showing significant increases in comparison with (54.48±35.58) pg/ml and (2 196.13±1 185.39) pg/ml in the POAG patients or (47.98±17.69) pg/ml and (1937.28±933.27) pg/ml in the ARC patients (aqueous:F =7.88,P＜0.01;plasma:F =4.18,P＜0.05).The concentration of IL-6 in aqueous humor and plasma were (234.87±41.64) pg/ml and (26.97±8.19) pg/ml from NVG patients,with considerable elavation in comparison with (38.97± 19.06)pg/ml and (19.54±5.11)pg/ml in the POAG patients or (29.48±14.61) pg/ml and (18.50±3.57) pg/ml in the ARC patients (aqueous:F =166.27,P＜0.01;plasma:F=5.59,P＜0.05).Conclusions The aqueous and plasma VEGF,TGF-β1 and IL-6 are considerably higher in NVG patients than those in POAG patients and ARC patients,suggesting that these cytokines participate in the pathogenesis and development of the NVG and probably are treating target of NVG.

BACKGROUND:Comeal graft endothelial cell density may decline spontaneously at a super-physiological speed after penetrating keratoplasty (PKP),even if no corneal rejection happens,and this is chronic corneal allograff dysfunction (CCAD).OBJECTIVE:To investigate the effects of topical corticosteroid treatment on chronic corneal allograft dysfunction after PKP.DESIGN,TIME AND SETTING:A controlled clinical analysis was performed at the Department of Ophthalmology,Tongji Hospital,Tongji University from May 2004 to May 2009.PARTICIPANTS:A total of 54 patients (eyes) who underwent PKP were included in this study.METHODS:Tobramycin-dexamethasone eyedrops was applied 6 times per day as an initial dosage and tapered during 4 months.Thereafter,drug withdrawal group and long term group were randomly divided.Treatments were ceased in the drug withdrawal group but were continued in the long-term group with a dosage of twice per day till 12 months after surgery.MAIN OUTCOME MEASURES:Corneal graft endothelial cell density was followed up.The mean follow up pedod was 3.12±0.28years.RESULTS:The corneal graft endothelial cell density of drug withdrawal and long term groups was dramatically declined within 36 months after PKP.At every time point,the endothelial cell density was significantly decreased compared to the last follow up (all P＜0.01 ).The endothelial cell density was decreased from (2 428±2.11) cells/mm~2 and (2 791±2.29) cells/mm~2 to (1 424±124) cells/mm~2 and (1 614±114) cells/mm~2 in the drug withdrawal and long-term groups,respectively.There was significant difference in endothelial cell density between the two groups no matter prior to surgery or at each time points after surgery,but the declination trends of two groups were basically the same,indicating that graft loss of endothelial cell density was not impacted by topical corticosteroid treatment after PKP (P＞0.05).CONCLUSION:Chronic corneal allograft dysfunction after PKP is independent of topical corticosteroid treatment.