AIM:To investigate the role and underlying mechanisms of NADPH oxidase 4(NOX4)/transient receptor potential channel subfamily C member 6(TRPC6)in the context of podocyte damage in diabetic nephropathya comprehensive investigative study was warranted.METHODS:(1)Male Sprague-Dawley rats were randomly divided in-to four distinct groups:a control group,a diabetic nephropathy group,a NOX4 inhibitor GKT137831-treated group,and a combined diabetic nephropathy with NOX4 inhibitor GKT137831-treated group,each consisting of 8～10 rats.The type 1 diabetes mellitus model was constructed via a single intraperitoneal injection of streptozotocin(STZ)(70 mg/kg),subse-quent to the successful induction of the model,GKT137831(at the dose of 5 mg/kg)was administered intraperitoneally.Regular monitoring of blood glucose levels was conducted,and urinary albumin excretion was quantified after 24 hours.Moreover,blood and kidney tissues were harvested for further analysis.(2)Mouse glomerular podocytes were divided into four distinct groups:a normal control group,a high glucose group,a GKT137831-treated group and a high glucose GKT137831-treated group.These podocytes were subsequently cultivated in vitro under high glucose conditions for 2 weeks.Thereafter,transfection of podocytes was carried out using NOX4 inhibitors and short interfering RNA targeting(siRNA)TRPC6.To detect the expression levels of NOX4 and TRPC6,a battery of techniques including Western blot,Immunohistochemistry,and reverse transcription polymerase chain reaction(RT-qPCR)were employed.(3)The morpho-logical changes of podocyte mitochondria under the condition of high glucose were observed by fluorescence confocal mi-croscopy,and the expression levels of peroxisome proliferator-activated receptor gamma coactivator 1α(PGC1α),mito-chondrial transcription factor A(TFAM),cytochromec oxidase subunit Ⅰ(COX Ⅰ)and cytochromec oxidase subunit Ⅳ(COX Ⅳ)in podocytes were assessed utilizing the Western blot technique.RESULTS:(1)Compared with normal con-trol group,mice with diabetic nephropathy manifested pronounced glomerular hypertrophy,thickening of basement mem-brane,expansion of the mesangial region,and an increased rate of urinary albumin excretion was observed.Analytical techniques such as Western blot and Immunohistochemistry showed a significant upsurge in the expression level of the NOX4 protein in kidney tissue,a diminished expression of glomerular podocyte protein(nephrin),an increased expression of interstitial cell markers(desmin),and an enhanced level of TRPC6 expression(P＜0.05).GKT137831 was assoiated with a reduction in desmin expression in renal tissue,preservation of glomerular nephrin expression,and a decrease in uri-nary albumin excretion(P＜0.05).(2)In vitro podocyte experiment,the expression of NOX4 and TRPC6 in podocyte was significantly increased in the context of high glucose(P＜0.05).Findings from Immunofluorescence and Western blot showed that GKT137831 effectively diminished the expression levels of TRPC6 and desmin while partially rescuing neph-rin expression in podocellular cells(P＜0.05).Western blot results showed that transfection of TRPC6 small interfering RNA could further promote the protective effect of GKT137831 on podiocytes.(3)Under high-glucose conditions,fluores-cence confocal microscopy revealed mitochondrial morphological damage in podocytes.However,therapeutic intervention with GKT137831 and transfection of TRPC6 siRNA partially rescued the mitochondrial structural integrity.Under high glucose conditions,immunoblot analysis demonstrated a marked decrement in the protein expression levels of PGC1α,TFAM,COX Ⅰ,and COX Ⅳ in podocytes.Importantly,GKT137831 and the transfection of TRPC6 siRNA significantly upregulated the levels of PGC1α,TFAM,COX Ⅰ,and COX Ⅳ(P＜0.05).CONCLUSION:In the context of the patho-genesis of diabetic nephropathy,the increased expression of NOX4 in the kidneys contributes to podocyte damage,with the effect being partially mediated via the TRPC6 channel.Inhibiting the NOX4-TRPC6 signaling pathway has the potential to ameliorate mitochondrial dysfunction in podocytes.This finding offers novel perspectives and strategies for the clinical di-agnosis and treatment of diabetic nephropathy.

AIM:To investigate the role and underlying mechanisms of NADPH oxidase 4(NOX4)/transient receptor potential channel subfamily C member 6(TRPC6)in the context of podocyte damage in diabetic nephropathya comprehensive investigative study was warranted.METHODS:(1)Male Sprague-Dawley rats were randomly divided in-to four distinct groups:a control group,a diabetic nephropathy group,a NOX4 inhibitor GKT137831-treated group,and a combined diabetic nephropathy with NOX4 inhibitor GKT137831-treated group,each consisting of 8～10 rats.The type 1 diabetes mellitus model was constructed via a single intraperitoneal injection of streptozotocin(STZ)(70 mg/kg),subse-quent to the successful induction of the model,GKT137831(at the dose of 5 mg/kg)was administered intraperitoneally.Regular monitoring of blood glucose levels was conducted,and urinary albumin excretion was quantified after 24 hours.Moreover,blood and kidney tissues were harvested for further analysis.(2)Mouse glomerular podocytes were divided into four distinct groups:a normal control group,a high glucose group,a GKT137831-treated group and a high glucose GKT137831-treated group.These podocytes were subsequently cultivated in vitro under high glucose conditions for 2 weeks.Thereafter,transfection of podocytes was carried out using NOX4 inhibitors and short interfering RNA targeting(siRNA)TRPC6.To detect the expression levels of NOX4 and TRPC6,a battery of techniques including Western blot,Immunohistochemistry,and reverse transcription polymerase chain reaction(RT-qPCR)were employed.(3)The morpho-logical changes of podocyte mitochondria under the condition of high glucose were observed by fluorescence confocal mi-croscopy,and the expression levels of peroxisome proliferator-activated receptor gamma coactivator 1α(PGC1α),mito-chondrial transcription factor A(TFAM),cytochromec oxidase subunit Ⅰ(COX Ⅰ)and cytochromec oxidase subunit Ⅳ(COX Ⅳ)in podocytes were assessed utilizing the Western blot technique.RESULTS:(1)Compared with normal con-trol group,mice with diabetic nephropathy manifested pronounced glomerular hypertrophy,thickening of basement mem-brane,expansion of the mesangial region,and an increased rate of urinary albumin excretion was observed.Analytical techniques such as Western blot and Immunohistochemistry showed a significant upsurge in the expression level of the NOX4 protein in kidney tissue,a diminished expression of glomerular podocyte protein(nephrin),an increased expression of interstitial cell markers(desmin),and an enhanced level of TRPC6 expression(P＜0.05).GKT137831 was assoiated with a reduction in desmin expression in renal tissue,preservation of glomerular nephrin expression,and a decrease in uri-nary albumin excretion(P＜0.05).(2)In vitro podocyte experiment,the expression of NOX4 and TRPC6 in podocyte was significantly increased in the context of high glucose(P＜0.05).Findings from Immunofluorescence and Western blot showed that GKT137831 effectively diminished the expression levels of TRPC6 and desmin while partially rescuing neph-rin expression in podocellular cells(P＜0.05).Western blot results showed that transfection of TRPC6 small interfering RNA could further promote the protective effect of GKT137831 on podiocytes.(3)Under high-glucose conditions,fluores-cence confocal microscopy revealed mitochondrial morphological damage in podocytes.However,therapeutic intervention with GKT137831 and transfection of TRPC6 siRNA partially rescued the mitochondrial structural integrity.Under high glucose conditions,immunoblot analysis demonstrated a marked decrement in the protein expression levels of PGC1α,TFAM,COX Ⅰ,and COX Ⅳ in podocytes.Importantly,GKT137831 and the transfection of TRPC6 siRNA significantly upregulated the levels of PGC1α,TFAM,COX Ⅰ,and COX Ⅳ(P＜0.05).CONCLUSION:In the context of the patho-genesis of diabetic nephropathy,the increased expression of NOX4 in the kidneys contributes to podocyte damage,with the effect being partially mediated via the TRPC6 channel.Inhibiting the NOX4-TRPC6 signaling pathway has the potential to ameliorate mitochondrial dysfunction in podocytes.This finding offers novel perspectives and strategies for the clinical di-agnosis and treatment of diabetic nephropathy.

Objective:To explore the clinical significance of the MYCN gene, PHOX2B gene and plasma cell-free DNA (cfDNA) in risk stratification and predicting the prognosis of high-risk neuroblastoma (NB). Methods:This was a prospective study involving 94 high-risk NB children admitted to Beijing Children′s Hospital, Capital Medical University from August 2017 to December 2018.Relative levels of MYCN and PHOX2B and cfDNA at diagnosis, and 4 and 6 cycles of chemotherapy were detected, and their differences were compared by the Chi- square test.Kaplan-Meier survival analysis was performed to explore their prognostic potential in high-risk NB. Results:Among the 94 high-risk NB children, 14 cases (14.9%) had MYCN amplification, 76 cases (80.8%) had positive expression of PHOX2B and 56 cases (59.6%) had cfDNA level higher than 100 μg/L.The proportion of high lactate dehydrogenase (LDH, ≥1 500 U/L) level in the MYCN gene amplification group (6/14 cases) was higher than that in the normal group (9/80 cases) ( P=0.009). The proportion of multi-site metastasis (54/76 cases) and high neuron specific enolase (NSE) level (NSE≥370 μg/L, 37/76 cases) in PHOX2B positive group were significantly higher than those in the negative group (5/14 cases, 2/14 cases) ( P=0.015, 0.020). The proportion of high LDH and high NSE in high cfDNA concentration (≥229.6 μg/L)group (13/37 cases, 28/37 cases) were significantly higher than those in low cfDNA concentration group (2/48 cases, 10/48 cases) (all P<0.001). With the decreased tumor burden during the treatment, the copy number of PHOX2B gene and cfDNA level were significantly lower than those at the initial diagnosis [0 (0-719.6) copies vs.1 723.5 (0-186 000.0) copies; 19.0 (1.1-225.5) μg/L vs.200.6 (8.0-5 247.4) μg/L, all P<0.001]. The 2-year event-free survival (EFS) rate of the MYCN gene amplification group was significantly lower than that of the normal group[(33.3±13.1)% vs.(58.5±7.1)%, P=0.020]. The 2-year EFS rate of PHOX2B positive group was significantly lower than that of the negative group[(47.9±7.1)% vs.(79.1±11.1)%, P=0.043]. EFS rate in high cfDNA concentration group was significantly lower than that in cfDNA low concentration group[(38.6±9.8)% vs.( 71.7±8.2)%, P=0.001]. After 6 cycles of chemotherapy, EFS rate in the PHOX2B positive group was significantly lower than that in the negative group [(16.7±14.4)% vs.( 60.6±6.6)%, P=0.014]; which was significantly lower in the Metaiodobenzylguanidine (MIBG) positive group than that of the negative group[(35.2±11.7)% vs.(65.8±7.1)%, P=0.037]. The MYCN gene and cfDNA concentration were not correlated with the prognosis of high-risk NB.Survival analysis of the combination of PHOX2B and MYCN gene ( PHOX2B+ /MIBG + , PHOX2B+ or MIBG + , PHOX2B-/MIBG -) showed a significant difference in the survival among three groups[0 vs.(53.6±1.2)% vs.(65.5±7.4)%, P=0.003]. Conclusions:The MYCN and PHOX2B gene and cfDNA concentration are of significance in risk stratification and predicting the prognosis of high-risk NB.Compared with the MYCN gene and cfDNA concentration, the PHOX2B gene is more suitable for monitoring the curative effect of chemotherapy on high-risk NB.A combined analysis of PHOX2B gene and MIBG before treatment can be more accurate in evaluating the treatment effect and residual lesions.

Objective:To summarize the clinical features of neuroblastoma (NB) with bone metastasis in infants and the prognostic factors.Methods:A retrospective analysis was performed on 32 patients aged ≤12 months who were enrolled in Beijing Children′s Hospital, Capital Medical University from January 2010 to December 2019 and had imaging findings suggesting signs of distant bone metastasis.The control group was included NB children, aged ≤12 months, who were admitted to Beijing Children′s Hospital, Capital Medical University during the same period, without signs of distant bone destruction.The clinical manifestations and auxiliary examinations of infants with bone metastasis were summarized, and the efficacy evaluation and survival analysis of infants with regular treatment and follow-up were conducted until December 31, 2020. Kaplan- Meier survival analysis was used for prognostic analysis, and Log Rank test was used for univariate prognostic analysis. Results:There were 32 NB infants with bone metastases, involving 12 males (37.5%) and 20 females (62.5%), accounting for 16.0% (32/200 cases) of infants diagnosed with NB du-ring the same period.The median age of onset was 9 (4.5-12.0) months.The main primary site included the retroperitoneal and adrenal region in 24 cases(75.0%) and mediastinum in 3 cases (9.4%). Among the 32 cases, 14 cases (43.8%) had simple bone metastasis, 19 cases (59.4%) had distant lymph nodes, 18 cases (56.3%) had bone marrow, and 3 cases (9.4%) had intracranial and meningeal metastasis.Bone metastasis mainly occurred in the skull, with 11 cases of single bone metastases and the remaining with 2 or more bone metastases.Compared with 168 NB infants without bone metastasis, the prognosis of those with bone metastasis was significantly worse [3-year overall survival(OS) rate 97.6% vs.82.7%, P=0.001]. Univariate analysis showed that the prognosis of NB children with bone marrow metastasis, meningeal and intracranial metastasis, MYCN gene amplification, and high-risk group was poor (all P<0.05). Two patients returned to the local hospital for treatment after diagnosis.A total of 30 children were recruited for efficacy evaluation and prognostic analysis.Twenty-nine children underwent surgery, of which 6 cases received surgery before chemotherapy and 23 cases received surgery after chemotherapy.One case received chemotherapy only.The mean course of chemotherapy was 6.2 (4-13) times.One case was treated with radiotherapy, 1 case was treated with Metaiodobenzylguanidine (MIBG) therapy, and 1 case was treated with stem cell transplantation.A total of 18 cases (62.1%) event-free survived, and 12 cases (40.0%) had a mean event at 7 (1.5-32.0) months.Among them, 7 cases survived and 5 cases died (16.7%). The expected 3-year event-free survival rate and OS rate were 57.1% and 82.7%, respectively. Conclusions:The most common sites of infant NB metastasis are bone and bone marrow, and the most common sites of bone metastasis are skull.Infants with bone metastasis had a worse prognosis than those without bone metastasis, and infants with bone and bone marrow metastasis had a worse prognosis than infants with single bone metastasis.

Objective:To investigate the clinical characteristics, treatment effect and prognosis of children with nearly diploid neuroblastoma (NB).Methods:A retrospective analysis of the general clinical characteristics (including age, Gender, risk grouping, location of primary tumor, etc.), laboratory test results, treatment and recent prognosis of NB children with nearly diploidy in bone marrow chromosomes by G-banding technology who admitted to Beijing Children′s Hospital, Capital Medical University from January 2015 to December 2018. Kaplan- Meier method was adopted to calculate survival rate.Univariate analysis was performed using Log- Rank test, and multivariate analysis was conducted with Cox regression model. Results:A total of 43 patients, including 27 males and 16 females, with diagnosis were included, with 14 cases in the hypodiploid group and 29 cases in the hyperdiploid group, and the median age was 35.5 months.The 43 children were all in the high-risk group of International Neuroblastoma Staging System(INSS)-Ⅳ.The primary tumors were mainly located in the retroperitoneal adrenal region (83.7%, 36/43 cases). The largest diameter of the tumors was more than 10 cm (53.5%, 23/43 cases), and often accompanied by 2 or more metastases at the time of consultation.In terms of chromosome karyotype and chromosome karyotype of 14 children in the hypodiploid group was 41-45, the most common karyotype was 45 chromosomes[9 cases(64.3%)]. Among 29 children in the hyperdiploid group of the 47 chromosome karyotypes, 11 cases were common (37.9%). Tumor markers were as follows: neuron enolase (NSE) increased in 41 cases children (95.3%) at first diagnosis, and 25 cases (58.1%)> 370 μg/L; 42 cases (97.7%)had lactate dehydrogenase (LDH). The LDH of children in the hypodiploid group was all> 500 U/L, with 1 case was> 10 000 U/L.Nine cases (20.9%) of MYCN gene were detected by fluorescence in situ hybridization (FISH). Treatment and prognosis: the total course of chemotherapy for 43 patients was 1-12, 19(44.2%) patients received autologous stem cell transplantation, 21 patients (46.5%) received postoperative or autologous radiotherapy or metaiodobenzylguanidine treatment, 28 children developed or relapsed with a median duration of 13.8 months, and 15 cases (34.9%) died.The median follow-up time of the 14 children in the hypodiploid group was 14.9 months (2-38 months), 12 cases progressed or relapsed, and 7 died.The median follow-up of 29 children in the hyperdiploid group was 20.0 months (8.1-51.6 months), with 16 patients progressed or relapsed and 8 cases died. Kaplan- Meier survival analysis illustrated that the 3-year projected event free survival (EFS) rate of 43 children was 18.4%, of which 17.1% were in the hypodiploid group and 29.8% in the hyperdiploid group. Conclusions:Preliminary analysis reveals that children with nearly diploid NB are mostly in the stage Ⅳ high-risk group over the age of 18 months, and 2 or more metastases at the time of consultation.The 3-year estimated EFS of 43 children was 18.4%, and the prognosis was worse in the hypodiploid group.

Objective:To summarize and analyze the results of chromosome karyotype in children with neuroblastoma (NB) with bone marrow metastasis at first diagnosis, and to discuss the clinical significance.Methods:G-banding was applied to the analysis of chromosome karyotype of patients who were regularly treated in the Hematological and Oncology Center in Beijing Children′s Hospital from January 2015 to December 2017, and all the patients were followed up until December 31, 2018.Their clinical features and prognosis were analyzed.Results:(1) There were 120 cases with bone marrow metastasis, including 74 boys and 46 girls, and 98 cases (81.7%) were ≥ 18 months.Among 60 cases with normal chromosome, 56 cases (93.3%) were in International Neuroblastoma Staging System(INSS)-Ⅳ phase, and 4 cases in INSS-Ⅳs phase; there were 2 low-risk (LR) cases, 9 intermediate-risk (MR) cases, and 49 high-risk (HR) cases (81.7%); 7 cases had MYCN gene amplifications.All 60 patients with chromosome abnormalities were in INSS-Ⅳ phase; there was 1 case in MR and 59 cases (98.3%) in HR; 14 cases had MYCN gene amplifications.(2) Among 60 children (50%) with chromosome abnormalities, 4 children had number abnormalities, 14 children had structural abnormalities, and 42 children had both number and structural chromosome abnormalities.Chromosome 21, 10, 11 deletions were the most common in number abnormalities; structural abnormalities involving 11q, 1p, 3p segments had a high incidence.(3) Seventeen cases of children with normal chromosome had tumor progression or recurrence during the 4 to 44-month follow-up period, and 31 cases of children with chromosome abnormalities had tumor progression or recurrence during the 2 to 42-month follow-up period.The 3-year overall survival rate and event-free survival rate of all children were 60.0% and 48.4%, respectively; children in the normal chromosome group had a 3-year overall survival rate of 74.2% and an event-free survival rate of 65.7%; the 3-year overall survival rate and event-free survival rate of children with chromosome abnormalities were 47.5% and 24.9%, respectively.Most children suffering from tumor progression or recurrence had chromosome 10 deletion, and abnormal structure of 11q, 1p, 2p segments. Conclusion:The chromosomal abnormality rate of Nb children's tumor cells is high, but the repetition rate is low, and the individual difference is obvious.The deletion of chromosome 10, abnormal regional structure of 11q, 1p and 2p segments may be poor prognostic factors for NB.Chromosome karyotype analysis of bone marrow samples is feasible, which can provide a basis for more accurate risk stratification and treatment.

The reflective ability of nursing staff has been paid more and more attention, nurses' reflection can promote professional competence development, which also can improve clinical practice ability, knowledge expansion ability and innovation ability. Therefore this article reviews the current situation on nurses′ reflection, the assessment tools, the modes, forms, methods and types, future prospects of clinical nurses reflective practice, in order to provide reference and basis for the relevant research on the reflection practice of clinical nursing staff in China.

Objective:To summarize the different teaching models and their effects in evidence-based medicine at home and abroad by qualitative method and systematic review.Methods:We searched the following databases (from inception to 13 May, 2019): PubMed, Embase, Proquest, Cochrane, Web of Science database and the Chinese databases (CNKI, Wanfang, SinoMed and VIP). To assess data strength and validity, risk of bias assessments were undertaken.Results:A total of 52 literatures were included in this study, including 21 Chinese-language literature and 31 English-language literature. PBL teaching model, mixed teaching model and workshop teaching model were the three teaching models with the largest number of studies in 20 teaching models.Conclusion:The evidence-based medicine teaching effect was closely related to the teaching models, so it is necessary to explore more suitable teaching models for the evidence-based medicine to improve the teaching effects.

Excess activation of cardiac fibroblasts inevitably induces cardiac fibrosis. Emodin has been used as a natural medicine against several chronic diseases. The objective of this study is to determine the effects of emodin on cardiac fibrosis and the underlying molecular mechanisms. Intragastric administration of emodin markedly decreased left ventricular wall thickness in a mouse model of pathological cardiac hypertrophy with excess fibrosis induced by transaortic constriction (TAC) and suppressed activation of cardiac fibroblasts induced by angiotensin II (AngII). Emodin upregulated expression of metastasis associated protein 3 (MTA3) and restored the MTA3 expression in the setting of cardiac fibrosis. Moreover, overexpression of MTA3 promoted cardiac fibrosis; in contrast, silence of MTA3 abrogated the inhibitory effect of emodin on fibroblast activation. Our findings unraveled the potential of emodin to alleviate cardiac fibrosis upregulating MTA3 and highlight the regulatory role of MTA3 in the development of cardiac fibrosis.

Objective To evaluate the curative effect of compound nutrient assisted phototherapy on neonatal jaundice.Methods Neonatologists at seven hospitals participated in the study.A total of three hundred and twenty full-term newborns with high indirect bilirubin admitted to hospital from September 2017 to January 2018 were selected.One hundred and sixty-six cases in the observation group,and one hundred and fifty-four cases in the control group,all enrolled neonates were given single-sided,conventional intensity phototherapy.Observation group took compound nutrient at the same time.The average gestational age,age,birth weight of two groups before treatment were not significantly different.Serum total biilirubin,indirect bidirubin,liver function (ALT,AST) and phototherapy time were monitored before treatment and 3 days after treatment.Results The serum total bilirubin in the observation group was significantly lower than that of the control group after 3 days of treatment[(196.7 ± 57.2) μmol/L vs (216.5 ± 54.6) μmol/L],(t=3.17,P＜0.01).The indirect bilirubin in the observation group was significantly lower than that of the control group after 3 days of treatment [(176.3 ± 54.3) μmol/L vs (197.2 ± 52.9) μmol/L],(t=3.50,P＜0.01).The time of phototherapy of the children in the observation group was significantly shorter than that of the control group[(19.8 ± 14.4)d vs (22.9 ± 13.3) d],(t =2.00,P ＜ 0.01).Rash,fever,bronze disease,spilled milk,vomiting,abdominal distention,diarrhea,constipation,liver damage etc.were no significant difference the observation group and the control group(P ＞ 0.05).Conclusion Compound nutrients had good efficacy and safety in adjuvant phototherapy for neonatal high indirect bidirubin.