Efficacy of plasma exchange in the treatment of autoimmune hemolytic anemia in children / 中国小儿急救医学

Objective:To investigate the efficacy and safety of plasma exchange(PE) in the treatment of autoimmune hemolytic anemia in children.Methods:The data from 8 hospitals in China during November 2014 to April 2017 were collected, and the clinical characteristics of PE in children with AHA were analyzed retrospectively.Results:A total of 21 children with AHA were included in the study, including 17 cases from PICU and 4 cases from pediatric kidney ward, with 11 boys and 10 girls, and the median age was 3.64(0.25, 11.10)years old, and median hospital stay was 12(4, 45)days.There were 15 cases(71.4%) with infection, 2 cases(9.5%)with autoimmune diseases, 4 cases(19.0%) with unknown.Consciousness disturbance occurred in 4 patients before replacement and recovered to normal after PE.The volume of blood decreased in two cases(9.5%) and completely relieved.There were 20 cases of anemia (95.2%), 15 cases were normal after PE, and 5 cases were improved.Jaundice occurred in 18 cases (85.7%), 12 cases were normal after PE, 6 cases were improved.Hepatosplenomegaly was found in 11 cases, 10 cases were normal after PE, 1 case was improved.After PE, the hemoglobin and red blood cell count increased, while the total bilirubin, indirect bilirubin, urea nitrogen and lactate dehydrogenase decreased, there were significant differences between pre-and post-replacement ( P<0.05). Only 1 case had allergic reaction, which was improved after symptomatic treatment, and PE was continued.After PE, 2 cases (9.5%) had complete remission, 16 cases (76.2%) had partial remission and 3 cases (14.3%) had been discharged. Conclusion:PE therapy can obviously improve the clinical symptoms and laboratory indexes of children with AHA who have failed to respond to conservative treatment.It can be used as a treatment measure for children with severe AHA and has a good safety.

Efficacy of plasma exchange in the treatment of autoimmune hemolytic anemia in children / 中国小儿急救医学

Objective:To investigate the efficacy and safety of plasma exchange(PE) in the treatment of autoimmune hemolytic anemia in children.Methods:The data from 8 hospitals in China during November 2014 to April 2017 were collected, and the clinical characteristics of PE in children with AHA were analyzed retrospectively.Results:A total of 21 children with AHA were included in the study, including 17 cases from PICU and 4 cases from pediatric kidney ward, with 11 boys and 10 girls, and the median age was 3.64(0.25, 11.10)years old, and median hospital stay was 12(4, 45)days.There were 15 cases(71.4%) with infection, 2 cases(9.5%)with autoimmune diseases, 4 cases(19.0%) with unknown.Consciousness disturbance occurred in 4 patients before replacement and recovered to normal after PE.The volume of blood decreased in two cases(9.5%) and completely relieved.There were 20 cases of anemia (95.2%), 15 cases were normal after PE, and 5 cases were improved.Jaundice occurred in 18 cases (85.7%), 12 cases were normal after PE, 6 cases were improved.Hepatosplenomegaly was found in 11 cases, 10 cases were normal after PE, 1 case was improved.After PE, the hemoglobin and red blood cell count increased, while the total bilirubin, indirect bilirubin, urea nitrogen and lactate dehydrogenase decreased, there were significant differences between pre-and post-replacement ( P<0.05). Only 1 case had allergic reaction, which was improved after symptomatic treatment, and PE was continued.After PE, 2 cases (9.5%) had complete remission, 16 cases (76.2%) had partial remission and 3 cases (14.3%) had been discharged. Conclusion:PE therapy can obviously improve the clinical symptoms and laboratory indexes of children with AHA who have failed to respond to conservative treatment.It can be used as a treatment measure for children with severe AHA and has a good safety.

Cell HE staining smears and paired cell paraffin sections in detection of epithelial growth factor receptor gene of pleural fluid specimens / 中南大学学报(医学版)

OBJECTIVES@#The advanced non-small cell lung cancer (NSCLC) patients with pleural effusion have no opportunity for surgery treatment. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the first-line drugs for these patients with EGFR-sensitive mutation. However, the disease progression and drug update during or after treatment of EGFR-TKIs bring more challenges and puzzles to clinical diagnosis and treatment, which inevitably requires archived pleural cell samples for EGFR re-examination or comparative study. Understanding the DNA quality of archived pleural fluid samples and effectively using archival data of pleural fluid cells are of great significance for tracing the origin of cases and basic medical research. This study aims to evaluate the consistency of EGFR mutant gene expression between the 2 methods, and to explore a reliable way for preserving cytological data and making full use of cytological archival data via cell HE staining smear and cell paraffin section.@*METHODS@#A total of 57 pleural fluid cytology cases in the Department of Pathology of China Aerospace Center Hospital from October 2014 to April 2021 were selected. Tumor cells were detected by cell HE staining smears and immunohistochemical staining for TTF-1 and Napsin A in the paired cell paraffin sections. There were more than 200 tumor cells in cell HE staining smear and the proportion of tumor cells were ≥70% in matched cell paraffin sections. Patients with 2 cell smears (one for cell data retention and the other for DNA extraction) were selected as the research subjects, and 57 pleural fluid samples were enrolled. EGFR gene mutation was detected by amplification refractory mutation system-polymerase chain reaction in 57 paired cell HE staining smears and cell paraffin sections. DNA concentration was 2 ng/μL. Cell HE smear was amplified side-by-side with DNA samples from paired cell paraffin sections. Result determination was according to the requirements of the reagent instructions. The external control cycle threshold (Ct) value of the No. 8 well of the samples to be tested was between 13 and 21, which was considered as successful and reliable samples. When the Ct value of EGFR gene mutation was <26, it was considered as positive; when the Ct value was between 26 and 29, it was critical positive; when the Ct value was equal or more than 29, it was negative. ΔCt value was the difference between mutant Ct value and externally controlled Ct value. The smaller the ΔCt value was, the better the quality of DNA of the detected sample was.@*RESULTS@#Among the 57 pleural effusion samples, 42 patients were hospitalized with pleural effusion as the first symptom, accounting for 73.7% (42/57). EGFR mutation was detected in 37 samples [64.9% (37/57)]. The mutation rate for 19del was 37.8% (14/37) while for L858R was 48.6% (18/37). Females were 56.7% (21/37) of mutation cases. The mutation consistency rate of cell HE staining smear and matched cell paraffin sections was 100%. The ΔCt values of cell HE staining smears were less than those of matched cell paraffin sections. The mutation Ct values of 37 cytological samples were statistically analyzed according to the preservation periods of the years of 2014-2015, 2016-2017, 2018-2019, and 2020-2021. There were significant differences in cell paraffin section in the years of 2014-2015 and 2016-2017 compared with the years of 2018-2019 and 2020-2021, while no significant differences were found in cell HE staining smear. Statistical analysis of externally controlled Ct values of 57 cytological samples showed that there were significant differences between cell HE staining smears and cell paraffin section in the years of 2014-2015 and 2016-2017, compared with the years of 2018-2019 and 2020-2021. The mutational Ct values of 37 paired cell blocks and smears were all <26, and the externally controlled Ct values of 57 paired cell paraffin sections and HE staining smears were all between 13 and 21.@*CONCLUSIONS@#The DNA quality of cell HE smears and matched cell paraffin section met the qualified requirements. Two methods possess show an excellent consistency in detecting EGFR mutation in NSCLC pleural fluid samples. The DNA quality of cell HE staining smear is better than that of cell paraffin sections, so cell HE staining smear can be used as important supplement of the gene test source. It should be noted that the limitation of cell HE staining smears is non-reproducibility, so multiple smears of pleural fluid are recommended to be prepared for multiple tests.

Reliability and validity of the theory of planned behavior psychological help-seeking scale in Chinese adults / 中华行为医学与脑科学杂志

Objective:To examine the reliability and validity of the theory of planned behavior psychological help-seeking questionnaire (TPB-PHSQ) in Chinese adults.Methods:A total of 979 Chinese adults were recruited through convenient sampling method.The final scale was determined by item analysis and exploratory factor analysis.Then, the TPB-PHSQ was examined for reliability and validity.Results:The theory of planned behavior psychological help-seeking scale included 14 items in four sections: attitude, subjective norm, perceived behavioral control and help-seeking intention.The internal consistency reliability of the TPB-PHSQ were 0.893, 0.711, 0.869, 0.918, and the retest reliability were 0.798, 0.713, 0.712, 0.729 for the 4 sections.The confirmatory factor analysis met the fit criteria for each indicator ( χ2/ df= 2.585, RMSEA=0.063, SRMR= 0.067, NFI=0.934, RFI=0.912, IFI=0.958, TLI=0.944, CFI=0.958). TPB-PHSQ was significantly correlated with validated variable, which the correlation coefficients of attitude, perceived behavioral control and the score of the attitude toward professional psychological help-short form were 0.474, 0.357, and the correlation coefficients of subjective norm, help-seeking intention and the score of intention of seeking counseling inventory were 0.432, 0.415 ( all P<0.01). Conclusion:The TPB-PHSQ has good reliability and validity, which can be used for relevant research and application.

The multiple mediating effects of health belief and physician trust on proactive personality and medical attitude in adults / 中华行为医学与脑科学杂志

Objective:To explore the influence of proactive personality on medical attitude and the mediating role of health belief and physician trust.Methods:A total of 703 adults were surveyed by proactive personality scale, Chinese adult health belief scale, Wake Forest physician trust scale and medical attitude scale from Octomber to November 2019.Descriptive statistics and correlation analysis were conducted by SPSS 23.0 software, and mediating effects were analyzed by SPSS macro program PROCESS 3.3.Results:(1)For all subjects, the score of proactive personality was(55.12±11.00), the score of health belief was(74.88±10.31), the score of doctor trust was(33.23±5.63), and the score of medical attitude was(56.55±13.07). Proactive personality was significantly positively correlated with health belief ( r=0.535), physician trust ( r=0.298) and medical attitude ( r=0.406) (all P<0.01), and health belief was significantly positively correlated with physician trust ( r=0.363) and medical attitude ( r=0.696) (both P<0.01). There was a significant positive correlation between physician trust and medical attitude ( r=0.452, P<0.01). (2) Proactive personality had an impact on medical attitude through three pathways. The single mediating effect value of health belief was 0.326, accounting for 82.74% of the total indirect effect. The single mediating effect value of physician trust was 0.040, accounting for 10.15% of the total indirect effect. The chain mediating effect value of health belief and physician trust was 0.028, accounting for 7.11% of the total indirect effect. Conclusion:Health belief and physician trust play multiple mediating roles in proactive personality and medical attitude in adults.

Case report of infant perianal necrotizing fasciitis / 中华实用儿科临床杂志

The clinical data of a child with perianal necrotizing fasciitis admitted to the Department of Pedia-trics of Guangzhou First People′s Hospital were retrospectively analyzed.The child, girl, more than 5 months old, the clinical features of the onset of fever and diarrhea, only 10 days after the onset, the child′s skin progressed from redness and swelling to perianal skin and soft tissue ulcers, fat liquefaction, visible rectum exposure.After surgical incision, thorough debridement and drainage and selection of sensitive antibiotics, the child recovered and was discharged.Perianal necrotizing fasciitis is a rare necrotizing soft tissue infection caused by a variety of bacterial infections.Because its early performance is difficult to distinguish, the symptoms are serious, and the mortality rate is high.It should been pain attention in clinical work.

Multiple mediating effects of stigma and loneliness between anxiety and attitude towards seeking professional psychological help among college freshmen / 中华行为医学与脑科学杂志

Objective:To explore the mediating effect of stigma and loneliness between anxiety and attitude towards seeking professional psychological help among college freshmen.Methods:A sample of 920 freshmen was recruited in the study and they were required to complete self-rating anxiety scale(SAS), attitudes toward seeking professional psychological help scale-short form(ATSPPH-SF), questionnaire of stigma for seeking professional psychological help(SSPPH), University of California, Los Angeles loneliness scale(UCLA). SPSS 24.0 was used for descriptive analysis and correlation analysis.SPSS macro program PROCESS 3.4 was used to analyze the mediating effect.Results:(1) The scores of anxiety, stigma, loneliness and attitude towards seeking professional psychological help of all subjects were (44.70±7.13), (19.14±6.79), (52.30±6.65) and (18.12±3.49). Anxiety was positively correlated with stigma and loneliness( r=0.170, 0.351, both P<0.01). Stigma was positively correlated with loneliness ( r=0.264, P<0.01). Attitude towards seeking professional psychological help was negatively correlated with anxiety, stigma and loneliness( r=-0.085, -0.459, -0.185, all P<0.05). (2) There were three paths that anxiety affected attitude towards seeking professional psychological help.The mediating effect size of stigma was -0.037, accounting for 75.51% of the total indirect effect.The mediating effect size of loneliness was -0.011, accounting for 22.45% of the total indirect effect.The chain mediating effect of anxiety and attitude towards seeking professional psychological help was -0.001, accounting for 2.04% of the total indirect effect. Conclusion:Stigma and loneliness play multiple mediating effects on the relationship between anxiety and attitude towards seeking professional psychological help.

Chinesization of the attitudes toward medical help-seeking scale and its reliability and validity / 中华行为医学与脑科学杂志

Objective:To translate the attitudes toward medical help-seeking scale (ATMHSS) and evaluate the reliability and validity of ATMHSS Chinese version in Chinese residents.Methods:Totally 769 residents were chosen through convenient sampling method.Based on item analysis and exploratory factor analysis, ATMHSS Chinese version was determined and its reliability and validity were examined.Results:Four main factors were extracted by exploratory factor analysis which were pro-action (10 items), non-fatalistic (10 items), med trust (6 items) and non-avoidant (4 items). Correlations between each dimension and the total score ranged from 0.641 to 0.839 and the correlations between each dimension were from 0.213 to 0.556.There was a significant positive correlation between ATMHSS and criterion variables ( r=0.406-0.696, all P<0.01). The confirmatory factor analysis showed the four-factor model fit the data well(χ 2=752.572, df=391, χ 2/ df=1.925, RMSEA=0.045, SRMR=0.058, CFI=0.902, GFI=0.903, IFI=0.903, TLI=0.891). The Cronbach's α coefficients of ATMHSS degreed from 0.726 to 0.879, the retest reliability of ATMHSS degreed from 0.622 to 0.736. Conclusion:The revised ATMHSS Chinese version shows good reliability and validity, which could be used as a tool for relevant research and application.

Clinical trial progress of Rituximab therapy in children with primary nephrotic syndrome / 中华实用儿科临床杂志

Primary nephritic syndrome (PNS) is the most common glomerular disease in children.Glucocorticoid alone or concurrent with immunosuppressive agents is the main treatment for various types of PNS.However, about 20% of children with PNS still show frequent relapse or steroid dependence during or after withdrawal of hormones and/or immunosuppressive agents.For these children, the long-term and high-dose use of hormones and/or immunosuppressive agents cause serious side effects and even intolerance.Additionally, there are 1%-3% children with PNS showing no response to hormones and all immunosuppressive agents, and they need new treatment regiments.In recent 15 years, there have been increasing clinical trials on the treatment of various types of PNS with Rituximab (RTX). The study results show that RTX can prolong the remission period of PNS and reduce the dose and side effects of hormones and immunosuppressive agents.Moreover, some randomized controlled trials revealed that RTX can be used as a potential first-line drug for hormone-dependent or frequently-relapsing PNS or a second-line treatment for hormone-resistant PNS.In this article, the latest clinical trial designs and results of RTX in the treatment of PNS in children in recent years were summarized, so as to provide reference for the clinical practice and research of RTX in the treatment of PNS.

Effect of Tacrolimus on the expression of α-actinin-4 in podocytes induced by Puromycin aminonucleoside / 中华实用儿科临床杂志

Objective@#To investigate the effects of Tacrolimus(FK506) and Puromycin aminonucleoside(PAN) on apoptosis and expression of α-actinin-4 mRNA and protein in mouse glomerular podocytes in order to explore the protective effect of FK506 on podocytes.@*Methods@#Mouse glomerular podocytes were cultured in vitro, and the control group, PAN group and FK506 group were established.After 8 h, 24 h and 48 h of treatment, the cell morphology was observed and the apoptosis rate was detected.The cells were collected by real-time PCR and Western blot was used to detect the mRNA and protein expression of α-actinin-4.@*Results@#The cell body area of the PAN group was significantly smaller than that of the control group, and the cell area of the FK506 group was larger than that of the PAN group.There was no significant difference in the rate of podocyte apoptosis between those groups at 8 h (all P>0.05). At 24 h and 48 h, the apoptotic rate of podocytes in PAN group[(8.21±0.41)%, (16.32±0.17)%] were significantly higher than those in the control group[(4.28±0.35)%, (6.27±0.28)%], and the differences were significant (all P<0.05). The apoptosis rate of podocytes in FK506 group[(6.26±0.24)%, (13.32±0.24)%] were significantly lower than those in PAN group, and the differences were significant (all P<0.05). At 8 h, there was no significant difference in the expression of α-actinin-4 mRNA and protein(all P>0.05). The expression of mRNA (2.42±0.21, 3.78±0.25) and protein(0.77±0.04, 1.22±0.10) in the PAN group was significantly higher than mRNA(1.50±0.22, 2.15±0.15) and protein(0.44±0.03, 0.83±0.07) in the control group at 24 h and 48 h, and the differences were significant (all P<0.01). The expression of mRNA (1.65±0.24, 1.70±0.32) and protein (0.52±0.05, 0.56±0.07) in FK506 group was significantly lower than that of PAN group, and the differences were significant (all P<0.05).@*Conclusions@#FK506 can effectively inhibit the damage of PAN on podocytes and stabilize the expression of α-actinin-4, which provides a basis for the clinical application of FK506 in the treatment of glomerular diseases.

Reliability and validity of the experiences in close relationships scale-the Chinese short version in Chinese adults / 中华行为医学与脑科学杂志

Objective To revise Experiences in close relationship scale-short form (ECR-S) and e-valuate the reliability and validity of Experiences in Close Relationship Scale-the Chinese Short version (ECR-CS) in Chinese adult population. Methods A sample of 1 572 Chinese adults were chosen through convenient sampling method to test the structural validity,criterion correlation validity,internal consistency reliability and test-retest reliability of ECR-CS. Results The results of confirmatory factor analysis showed that ECR-CS had a two-factor structure and data fitted this two-factor structure well,χ2/df=3. 57,CFI=0. 95,GFI=0. 95,AGFI=0. 92,TLI=0. 92,NFI=0. 93,RMSEA=0. 07,SRMr=0. 04. In ECR-CS,the Cronbach's alpha coefficients for attachment anxiety and attachment avoidance were respectively 0. 85 and 0. 88,the test-retest reliability with an interval of two weeks were respectively 0. 75 and 0. 80. With reference to relevant studies,self-esteem scale(SES),social avoidance and distress scale(SADS) and depression,anxi-ety and stress scale(DASS) were selected as the criterions to test the criterion-related validity of ECR-CS. Attachment anxiety and attachment avoidance of ECR-CS were significantly positively correlated with social avoidance,social distress,depression,anxiety and stress(r=0. 20-0. 43,P<0. 01),and negatively correlated with self-esteem (r=-0. 43--0. 40,P<0. 01),indicating that ECR-CS had a good criterion-related validity. Conclusion ECR-CS has good reliability and validity,which is suitable for the research of adult attachment in Chinese adult population.

Effect of Tacrolimus on the eXpression of α-actinin-4 in podocytes induced by Puromycin aminonucleoside / 中华实用儿科临床杂志

Objective To investigate the effects of Tacrolimus( FK506 )and Puromycin aminonucleoside ( PAN)on apoptosis and expression of α-actinin-4 mRNA and protein in mouse glomerular podocytes in order to ex-plore the protective effect of FK506 on podocytes. Methods Mouse glomerular podocytes were cultured in υitro,and the control group,PAN group and FK506 group were established. After 8 h,24 h and 48 h of treatment,the cell mor-phology was observed and the apoptosis rate was detected. The cells were collected by real-time PCR and Western blot was used to detect the mRNA and protein expression of α-actinin-4. Results The cell body area of the PAN group was significantly smaller than that of the control group,and the cell area of the FK506 group was larger than that of the PAN group. There was no significant difference in the rate of podocyte apoptosis between those groups at 8 h( all P>0. 05). At 24 h and 48 h,the apoptotic rate of podocytes in PAN group[(8. 21 ± 0. 41)%,(16. 32 ± 0. 17)%]were significantly higher than those in the control group[(4. 28 ± 0. 35)%,(6. 27 ± 0. 28)%],and the differences were significant(all P<0. 05). The apoptosis rate of podocytes in FK506 group[(6. 26 ± 0. 24)%,(13. 32 ± 0. 24)%] were significantly lower than those in PAN group,and the differences were significant(all P<0. 05). At 8 h,there was no significant difference in the expression of α -actinin -4 mRNA and protein( all P >0. 05 ). The expression of mRNA(2. 42 ± 0. 21,3. 78 ± 0. 25)and protein(0. 77 ± 0. 04,1. 22 ± 0. 10)in the PAN group was significantly higher than mRNA(1. 50 ± 0. 22,2. 15 ± 0. 15)and protein(0. 44 ± 0. 03,0. 83 ± 0. 07)in the control group at 24 h and 48 h,and the differences were significant(all P<0. 01). The expression of mRNA(1. 65 ± 0. 24,1. 70 ± 0. 32)and protein(0. 52 ± 0. 05,0. 56 ± 0. 07)in FK506 group was significantly lower than that of PAN group,and the differ-ences were significant(all P<0. 05). Conclusions FK506 can effectively inhibit the damage of PAN on podocytes and stabilize the expression of α-actinin-4,which provides a basis for the clinical application of FK506 in the treat-ment of glomerular diseases.

Revision of attitudes toward seeking professional psychological help scale-short form / 中华行为医学与脑科学杂志

Objective To revise the attitudes toward seeking professional psychological help scaleshort form(ATSPPH-SF) and to exam the reliability and validity of ATSPPH-SF Chinese revision.Methods Based on studying and analyzing foreign related questionnaires,1 657 adults from Tianjin were chosen as subjects to exam the reliability and validity of ATSPPH-SF Chinese revision.Results The three-dimensional structure were obtained according to the results of exploratory factor analysis,and the total variance explained amounted to 54.03％.The results of Confirmatory factor analysis showed that the measurement model fit the data well (x2/df=2.94,RMSEA =0.04,CFI =0.97,GFI =0.98,SRMR =0.01,AGFI =0.97,TLI =0.95,IFI =0.97,NFI =0.96).The cronbach's α coefficients of the questionnaire degreed from.0.71 to 0.78.The retest reliability of the questionnaire degreed from 0.65 to 0.82.Correlation between each item and the total score ranged from 0.32 to 0.63 and the correlations between each dimension and total questionnaire were from 0.67 to 0.82.There was a significant positive correlation with ATSPPH(r=0.67,P＜0.01).Conclusion The revised ATSPPH-SF has good reliability and validity,which can satisfy the domestic demand for relevant research and application.

Clinical analysis of angiotensin-converting enzyme inhibitor in treatment of Alport syndrome / 中华实用儿科临床杂志

Objective To investigate the clinical efficacy and safety of angiotensin-converting enzyme inhibitor (ACEI) and angiotensin receptor blocker (ARB) in treatment of children with Alport syndrome (AS).Methods A total of 22 children with AS in Department of Pediatrics,Guangzhou First People's Hospital and Department of Pediatrics,Shenzhen People's Hospital between January 2013 and December 2017 were selected.But four children were not included in this study since they did not take medication regularly,and the other 18 cases were included in this study.All the 18 children were initially treated with ACEI.The observation time was from 1 to 5 years.If the symptoms were not effectively controlled,treatment plan would be changed to ACEI combined with ARB treatment.The observation time was from 1 to 3 years.The clinical data and laboratory examination results [including 24-hour urine protein (mg/24 h),urine red blood cell count,plasma albumin (Alb),urea nitrogen (BUN),serum creatinine (Scr),total cholesterol (TC)] were collected for retrospective analysis.Results Eighteen patients started their treatment with ACEI inhibitors (Fosinopril).Within 2 years of treatment,the urinary protein and urinary red blood cells in the children decreased to 47.7％ and 41.3％,respectively,and the differences were all statistically significant (all P ＜ 0.05),and the renal function was stable within the normal range.Two years later,7 patients had elevation of urinary protein and urinary erythrocyte elevations and decrease of renal function,and they were treated with ACEI and ARB (Losartan).The other children had no significant change in urine protein and urine red blood cells in the 3 rd,4th,and 5th year,and their renal function was stable.After ACEI treatment alone for 5 years,urinary protein was 47.8％ lower than before treatment,and the difference was statistically significant (P ＜ 0.05);urinary red blood cells decreased to 32.0％ compared with before treatment,and the difference was statistically significant (P ＜ 0.05).Seven patients with ACEI alone had poor efficacy,after the treatment with ACEI combined with ARB,the urinary protein and urine red blood cells were lower in the first year and the renal function improved.There was no significant change in urinary protein and urine red blood cells in the 2nd and 3rd year,and renal function was stable.After ACEI combined with ARB treatment for 3 years,urinary protein decreased to 42.3％ before treatment,and the difference was statistically significant (P ＜ 0.05),and urinary red blood cells decreased to 46.9％ compared with before treatment,and the difference was statistically significant(P ＜ 0.05).Conclusions ACEI treatment of children with AS can reduce urine protein and help delay renal failure.For children with poor efficacy of ACEI treatment,ACEI combined with ARB may have a certain effect.ARB can be used as an adjunctive treatment for patients with AS who have a poor response to ACEI alone.

Tacrolimus inhibits Puromycin induced injury of podocytes by stabilizing the expression and distribution of podocin / 中华实用儿科临床杂志

Objective To investigate the effect of Puromycin(PAN)and Tacrolimus(FK506)on the expre-ssion of podocin in podocytes,and discuss the mechanism of FK506 in improving proteinuria caused by the damage of glomerular podocytes.Methods Mouse podocyte were cultured and divided into 3 groups,which were control group, PAN group and FK506 group,respectively.The changes of each group after 8 hours,24 hours and 48 hours of treatment were detected by using phase-contrast microscope,and the expression and distribution of protein and mRNA were tested by adopting Western blot,quantitative real-time PCR and immunofluorescence technique.Results Compared with the control group(8.54 ± 0.25,8.27 ± 0.07,7.45 ± 0.13)at each time point(8 hours,24 hours and 48 hours),the soma size of the PAN group(6.41 ± 0.22,4.96 ± 0.09,3.76 ± 0.16)was reduced.But in the FK506 group(7.67 ± 0.06, 6.62 ± 0.04,5.57 ± 0.27),it was increased at each time point(8 hours,24 hours and 48 hours)compared with the PAN group.The podocytic process and the intercellular connection disappeared,and the distribution was significantly scattered.The mRNA(0.87 ± 0.15,0.78 ± 0.15,0.58 ± 0.12)and protein(0.82 ± 0.02,0.62 ± 0.03,0.50 ± 0.02) expressions of podocin increased in FK506 group and mRNA(0.63 ± 0.12,0.56 ± 0.01,0.48 ± 0.02),protein (0.71 ± 0.03,0.46 ± 0.01,0.34 ± 0.02)were observably reduced in PAN group at each time point(8 hours,24 hours and 48 hours)and showed abnormal distribution in PAN group,compared with the control group[podocin mRNA (1.22 ± 0.15,1.18 ± 0.06,0.87 ± 0.30),protein(0.86 ± 0.03,0.87 ± 0.03,0.61 ± 0.07)].Conclusions PAN attenuates the mRNA and protein expression of podocin by damaging podocytes,inversely,while FK506 protects poto-cyte injury by stabilizing the mRNA and protein expression of podocin,which maybe involve inhibition of proteinuria.It can be used as a target for the study and treatment of kidney diseases.

Inhibitory of Dexamethasone on podocytes apoptosis induced by Puromycin via stabilizing phosphatidylinositol 3 kinase/protein kinase B signaling pathway / 中华实用儿科临床杂志

Objective To investigate the role of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) signaling pathway in Dexamethasone (DEX) inhibiting podocytes apoptosis which was induced by Puromycin (PAN).Methods Mouse glomerular podocytes were cultured in vitro,and were divided into control group,dimethyl sulfoxide (DMSO) group,PAN group,DEX group,and LY294002 (inhibitor of PI3 K) group.The mRNA expression of CD2-associated protein (CD2AP) was measured by using real time fluorescent quantitative polymerase chain reaction,and intracellular distribution was detected by using indirect immunofluorescence staining.Co localization of CD2AP and p85 was detected by using confocal fluorescence microscopy.The expressions of Akt,phosphorylated (p)-Akt,glycogen synthase kinase-3β (GSK3 β) and phosphorylated (p)-GSK3β were evaluated by using Western blot.Results The expressions of CD2AP mRNA in PAN group at each time point (8 h,24 h,48 h) (1.11 ± 0.16,0.78 ±0.09,0.56 ± 0.43) were significantly lower than those in the control group (1.90 ± 0.26,2.09 ± 0.12,2.28 ±0.95),and the differences were statistically significant (all P ＜ 0.05);CD2AP distributed in foot process with uniform filament and discontinuous coarse particle around perinuclear;CD2AP and p85 distributed in cell membrane and cytoplasm evenly in control group,but accumulated in nuclei in the PAN group.The expressions of CD2AP mRNA in DEX group at each time point (8 h,24 h,48 h) (1.53 ± 0.14,2.15 ± 0.27,2.13 ± 0.15) were significantly higher than those in the PAN group,and the differences were also statistically significant (all P ＜ 0.05);the distribution density and range of CD2AP were greater than those in the PAN group,and the accumulation with p85 in nuclei decreased obviously.The expressions of p-Akt and p-GSK3β were inhibited by PAN in a dose-dependent manner (P ＜0.05).The expressions of p-Akt and p-GSK3 β were lowest after PAN stimulated at 15 min and 30 min respectively.However,the expressions of p-Akt and p-GSK3 β increased depending on the concentration of DEX (P ＜ 0.05).In addition,the expressions of p-Akt and p-GSK3 β could be blocked by LY294002 (P ＜ 0.01).Conclusion DEX can protect podocytes and inhibit podocytes apoptosis through stabilizing the expression and distribution of CD2AP.The stale expression of PI3K/Akt signaling pathway is the key factor in DEX protecting podocytes.

Clinical analysis of children having primary nephrotic syndrome complicated with posterior reversible encephalopathy syndrome on treatment of immunosuppressants / 中华肾脏病杂志

Objective To explore the relationship between posterior reversible encephalopathy syndrome (PRES) and the treatment of immunosuppressants such as cyclosporine A (CsA) and tacrolimus (FK506) in children with nephrotic syndrome.Methods The clinical data of nephrotic syndrome children with PRES caused by immunosuppressants who were hospitalized in Guangzhou First People's Hospital from June 2014 to May 2017 were collected.Their clinical characteristics,imaging features,treatments and prognosis were analyzed.Results A total of 23 children were enrolled,including 13 children with CsA and 10 children with FK506.In the concurrent of PRES 20 cases were in the activity stage of nephrotic syndrome,with large amounts of urinary protein,obvious edema,hypoalbuminemia and hyperlipidemia;while 3 cases were in the remission of nephrotic syndrome.The main clinical symptoms of PRES were hypertension,headache,epileptic attack,consciousness disorder,visual disorder and so on.Sixty-nine point six percent of children were using high dose immunosuppressive agents,and 78.3％ had high drug concentration.The cranial magnetic resonance imaging (MRI) results of 17 patients showed that they had T1 weighted (T1WI) hypointense,T2 weighted (T2WI) and fluid-attenuated inversion recovery (FLAIR) images hyperintense,as well as iso-and slight hypointense of diffusion-weighted image (DWI) in parietal-occipital regions or complicated with frontal lobes or basal nuclei region.Computer tomography (CT) examinations of 6 cases showed low-density focus of the occipital lobes.Children were relieved muscular spasm,debased intracranial hypertension,improved circulation,discontinued or reduced immunosuppressants at the onset of PRES.After these treatments,21 patients' symptoms and signs disappeared within one week;two patients suffered convulsions 2 times in one week,but recovered after one month.After three months 5 children had MRI and CT re-examination and it showed that their brain lesions disappeared.Conclusions PRES may be related to the dose and blood concentration of immunosuppressive agents.The immunosuppressants for nephrotic syndrome children should be increased gradually with low initiating doses.Physicians need to be precautious to prevent the occurrence of PRES once neurological symptoms occur.

The relationship between autophagy and renal tissue injury in adriamycin nephropathy rats / 中华肾脏病杂志

Objective To observe the formation of autophagosome,the expression and distribution of autophagy-related protein LC3-Ⅰ,LC3-Ⅱ and Beclin-1 in adriamycin nephropathy rats at different pathological periods,to explore the relationship between autophagy and renal tissue injury,the occurrence of proteinuria,the progression of renal disease.Methods Sixty normal male SD rats were randomly divided into control group (n=30) and model group (n=30),the rats in model group were injected with adriamycin(6.5 mg/kg) via tail-vein for one time,while the rats in control group were injected with saline.Urine protein quantitation of 24 hour,the levels of serum albumin and total cholesterol were measured serially at the 2,4,6,8,10 weeks.The changes of kidney tissue pathology were detected after HE,PAS and Masson staining by light microscope.The formation of autophagy were detected by transmission electron microscopy,the localization and distribution of LC3-Ⅰ,LC3-Ⅱ and Beclin-1 were detected by indirect immunofluorescence staining in kidney tissue,the autophagy-related proteins LC3-Ⅰ,LC3-Ⅱ and Beclin-1 expression was detected by Western blotting.Results In model group,urinary protein began to increase at the first two weeks,serum albumin decreased at the same time,and total cholesterol increased in the four weeks.There was a statistically significant difference compared with the control group (P ＜ 0.01).The Scr and BUN were increased slightly at the four weeks in model group,and showed the deterioration of renal function after the eight weeks.There was a statistically significant difference compared with the control group (P ＜ 0.01).Mesangial cell proliferation,mitochondrial swelling and foot process broadening and integration appeared early in the model group,while foot process disappearing and nuclear pyknosis were observed in the late by transmission electron microscope;Renal pathology gradually changed from mesangial proliferation to focal segmental glomerulosclerosis (FSGS) by light microscope.A low expression of autophagy was detected in renal tissue of control group rats by transmission electron microscopy and immunofluorescence microscope;in model group,with the progression of disease,the autophagy was significantly enhanced and maintained at a high level.With the progression of disease,the autophagyrelated proteins LC3-Ⅰ,LC3-Ⅱ and Beclin-1 was significantly enhanced in the model group than the control group (P＜0.05).Conclusion Autophagy is involved in renal tissue injury and the occurrence of proteinuria,closely related to the progression of renal disease.

Influence of dexamethasone on the expression and distribution of transient receptor potential cation channel 6 in glomerular podocytes / 中华肾脏病杂志

Objective To observe the changes of foot processes,expression and distribution of transient receptor potential cation channel 6 (TRPC6) in podocytes by puromycin aminonucleoside (PAN) and dexamethasone (DEX) intervention,then to investigate the function of TRPC6 in podocytes and its relation to proteinuria in kidney diseases.Methods Podocytes cultured in vitro were divided into three group:control group,PAN stimulation group and DEX intervention group.Mouse podocyte cell line (MPC5) were cultured in 0.02％ dimethyl sulfoxide (DMSO) in control group,subjected to PAN (50 μg/ml) treatment alone or with DEX (1 μmol/L) in other two groups for 8 h,24 h,48 h.The podocyte morphology was observed and took pictures by phase-contrast microscope,then the differences of morphology and areas were analyzed.The distribution,mRNA expression and protein expression of TRPC6 were detected by indirect immunocyto-fluorescence,real-time quantitative PCR and Western blotting,respectively.Results The well-developed podocyte arborization and interconnection was formed in control group,but PAN led to significant shrinkage of podocytes (P ＜ 0.05),together with podocyte foot process retraction,effacement and loss of cell contact.DEX significantly prevented the shrinkage and apoptosis of podocytes.The apoptosis rate was significantly increased after PAN stimulated 48 h (P ＜ 0.05).Real-time quantitative PCR and Western blotting found TRPC6 mRNA and protein expression were prone to increase in PAN group compared with control group (P ＜ 0.05).The distribution of TRPC6 becamed abnormal in PAN group.DEX decreased TRPC6 mRNA and protein expression at 48 h compared with PAN group (P ＜ 0.05).The abnormal distribution of TRPC6 was also alleviated by the protection of DEX.Conclusion DEX exerts a direct action to podocyte which retains the integrity of slit diaphragm against podocyte injury,and alleviates proteinuria via stabilizing mRNA,protein expression and distribution of TRPC6.

Distribution and expression of α-actin-4 in puromycin aminonueleoside injured mouse podocyte cell line / 中华实用儿科临床杂志

Objective To observe the expression and distribution of α-actin-4 mRNA through puromycin aminonueleoside(PAN) injury podocyte,and discuss the relation between α-actin-4 and podocyte damage.Methods Podocytes were cultured in vitro,and 2 groups were set up:control group and PAN stimulation group.The control group was cultured with concentration of 100 mL/L FBS RPMI 1640 nutrient solution culture,while the PAN group was cultivated to PAN(50 mg/L) treatment,and cell morphology,extraction of total RNA of α-actin-4 were observed in 8 h,24 h and 48 h.The podocyte morphology was observed and pictures were taken through phase-contrast microscope,then the differences of morphology and areas between the 2 groups were analyzed.The distribution and mRNA expression of α-actin-4 were detected by indirect immunocyto-fluorescence and real-time quantitative PCR,respectively.Results The well-developed podocyte arborization was formed after the in vitro induction,and the PAN treatment led to the podocyte foot process retraction and effacement together with the mouse podocyte cell line shrinkage and the loss of cell contact.The above time point α-actin-4 mRNA expressions between the 2 groups were compared,and there was no significant difference in 8 h (P ＞ 0.05),but significant difference was found in 24 h,48 h,α-actin-4 higher mRNA expression,with statistical significance(all P ＜0.01).α-actin-4 in the control group had thin filaments evenly distributed in the cytoplasm,but a radioactive distribution in foot process.In the experimental group,α-actin-4 pressure silk fiber was shorter,with disordered arrangement,and PAN stimulus after 24 h,α-actin-4 distribution in cytoplasm was decreased significantly,while cytoplasmic distribution was missing after 48 h.Conclusions The abnormal of distribution and mRNA expression of α-actin-4 is time-related to the PAN injury podocyte,and α-actin-4 is an important part of podocyte damage mechanism.