Objective:To examine the plasma expression levels and clinical significances of microRNA(miR)-101-3p and miR-141-3p in children with sepsis.Methods:One hundred and fifty-three children with sepsis admitted in Sanya People′s Hospital from January 2016 to October 2019 were divided into sepsis without shock group (94 cases) and septic shock group (59 cases). In addition, they were further divided into survival group (107 cases) and death group (46 cases) according to the 28-day survival.Another 60 healthy children were selected as the healthy control group.Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was performed to detect plasma levels of miR-101-3p and miR-141-3p in all subjects.Receiver operating characteristic curve(ROC) were depicted to identify the diagnostic and prognostic potentials of plasma miR-101-3p, miR-141-3p and procalcitonin(PCT) in sepsis. Pearson′ s correlation analysis was performed to analyze the correlation between the expression levels of miR-101-3p, miR-141-3p and PCT with Acute Physiology and Chronic Health Evaluation Ⅱ(APACHE Ⅱ)score, Sequential Organ Failure Assessment(SOFA)score, leukocyte count and C-reactive protein level in children with sepsis. Results:Plasma levels of miR-101-3p, miR-141-3p and PCT in septic shock group and sepsis without shock group were significantly higher than those in the healthy control group (all P<0.001). Moreover, plasma levels of miR-101-3p (4.25±1.46 vs.1.86±0.75), miR-141-3p (3.17±1.08 vs.1.20±0.52) and PCT [(20.75±9.36) μg/L vs.(5.80±2.40) μg/L] in septic shock group were significantly higher than those in sepsis without shock group (all P<0.001). In addition, plasma levels of miR-101-3p, miR-141-3p and PCT in survival group and death group were significantly higher than those in the healthy control group (all P<0.001). Notably, plasma levels of miR-101-3p (4.83±1.62 vs.1.40±0.58), miR-141-3p (3.50±1.13 vs.0.96±0.47), and PCT [(26.30±11.72) μg/L vs.(3.25±2.16) μg/L] in death group were significantly higher than those in the survival group (all P<0.001). ROC curve analysis showed that the area under the curve (AUC) and 95% confidence interval (95% CI) of the combined diagnosis of sepsis with miR-101-3p, miR-141-3p and PCT were significantly higher than that of miR-101-3p, miR-141-3p or PCT alone [0.908 (0.850-0.970) vs.0.810 (0.748-0.873), 0.784 (0.723-0.844) and 0.825 (0.764-0.883), respectively; Z1=4.682, Z2=5.380 and Z3=4.417, all P<0.05]. The sensitivity and specificity of the combined diagnosis was 92.5% and 84.0%, respectively.The AUC and 95% CI of the combined prediction of miR-101-3p, miR-141-3p and PCT in the mortality of children with sepsis children with were significantly higher than those with miR-101-3p, miR-141-3p or PCT alone [0.930 (0.872-0.986) vs.0.848 (0.786-0.907), 0.792 (0.730-0.853) and 0.820 (0.762-0.878), respectively; Z1=4.537, Z2=5.728 and Z3=5.106, all P<0.05]. The sensitivity and specificity of the combined prediction in the mortality was 94.6%, and 87.0%, respectively.Correlation analysis showed that miR-101-3p and miR-141-3p levels were positively correlated with PCT ( r=0.804, 0.773, all P<0.001), APACHE Ⅱ score ( r=0.738, 0.695, P<0.001) and SOFA score ( r=0.752, 0.764, all P<0.001). Conclusions:Plasma levels of miR-101-3p and miR-141-3p in children with sepsis significantly increased, which are correlated with the severity of sepsis.A combination detection of miR-101-3p, miR-141-3p and PCT has high diagnostic and prognostic potentials in children with sepsis.

Objective To explore the clinical features and expression of PLA2R in renal tissue of children with idiopathic membranous nephropathy.Methods Retrospective study was performed in patients with membranous nephropathy diagnosed through renal biopsy and the follow-up time was at least half a year in Shanghai Children's Hospital from January 2010 to February 2017.We compared their clinicopathological and pathological findings of IMN.Indirect immunofluorescence assay was used to detect glomerular PLA2R expression.We analyzed the differences of clinical features between the PLA2R negative and positive groups.T test,rank-sum test and Fisher exact test were used.Results Eleven cases had hematuria and proteinuria,9 cases presented with nephrotic syndrome,and 2 cases showed isolated proteinuria.Of the 22 cases of children with IMN,16 patients had complete remission (complete remission rate was 72.8％),and 22 patients had partial remission.The renal functiou of all cases was normal and in all cases the estimated glomerular filtration rate was ＞ 90 ml/(min· 1.73m2).Of 22 cases with IMN,7 cases were PLA2R-positive in renal tissue and 15 cases were PLA2R-negative.The age of positive group(10 years old) was older than the negative group (6 years old)(Z=-2.483,P＜0.05) and the time of positive group (6 months) for urine protein to return to negative was longer than the negative group (2.5 months) through treatment.These differences were significantly different (Z=-2.072,P＜0.05).Conclusions Hematuria and proteinuria can be found in most children with idiopathic primary membranous nephropathy.Prednisone combined with immunosuppressant was effective.The positive rate of PLA2R in renal tissue of children with IMN was about 32％.The age of PLA2.R positive group was older than the negative group.And the time of urine protein turning to negative in positive group was longer than that in the negative group.

Next generation sequencing (NGS) has developed very rapidly in the last decade. Compared with Sanger sequencing, NGS has the advantages of high sensitivity and high throughput. Movement disorders are a common type of neurological disease. Although traditional linkage analysis has become a standard method to identify the pathogenic genes in diseases, it is getting difficult to find new pathogenic genes in rare Mendelian disorders, such as movement disorders, due to a lack of appropriate families with high penetrance or enough affected individuals. Thus, NGS is an ideal approach to identify the causal alleles for inherited disorders. NGS is used to identify genes in several diseases and new mutant sites in Mendelian movement disorders. This article reviewed the recent progress in NGS and the use of NGS in Mendelian movement disorders from genome sequencing and transcriptome sequencing. A perspective on how NGS could be employed in rare Mendelian disorders is also provided.
Alleles ; Genetic Linkage ; High-Throughput Nucleotide Sequencing ; methods ; Humans ; Movement Disorders ; diagnosis ; genetics ; Sequence Analysis, DNA ; Transcriptome

OBJECTIVETo analyze the clinical features and gene mutation of Chinese children with Alport syndrome(AS).

METHODFrom May 2011 to May 2014, clinical and pathological information gathered from 25 patients was retrospectively analyzed. COL4A5, COL4A4 and COL4A3 genes were analyzed using next-generation sequencing in these patients, and gene mutations of related family members were identified by Sanger method.

RESULTOf these 25 cases, 19(76%) had X-linked Alport syndromes (XL-AS), 6 had autosomal recessive Alport syndromes (AR-AS). Twenty five patients had an onset of hematuria and proteinuria and in 8 cases the disease was induced by upper respiratory tract infections. Hearing loss was present in 2 of 25 (8%) cases and ocular lesions in 1 of 25 (4%). Renal pathology showed that 16 of them had minimal change disease (MCD), 8 mesangial proliferative glomerulonephritis (MsPNG), 1 focal segmental glomerulo-sclerosis (FSGS). Extensive lamination and split of glomerular basement membrane (GBM) dense layers were found in 2 (8%) of 25 patients. Twenty one of 25 patients (84%) showed abnormal renal α-chain distribution. COL4A5, COL4A4 and COL4A3 genes of 25 patients (23 families) were analyzed and 24 pathogenic mutations were identified: 18 in COL4A5, 1 in COL4A3 and 5 in COL4A4. It was observed that 13 patients inherited the mutation from the mother, 3 patients inherited from the father, 2 patients inherited 1 mutation from the mother and another mutation from the father, and 7 patients carried the novel mutations.

CONCLUSIONXL is the main inherited type in AS. Most of patients showed MCD and MsPNG in renal biopsy. This research examined 24 mutations and 16 mutations were not reported previously.

Child ; Deafness ; Genes, Recessive ; Genotype ; Hematuria ; Humans ; Kidney ; Mutation ; Nephritis, Hereditary ; genetics ; pathology ; Pedigree ; Phenotype

Objective To promote the diagnostic accuracy of intrapancreatic accessory spleen (IPAS).Methods The clinical data of 10 cases of IPAS admitted in Fudan University Zhongshan Hospital from Apr 2005 to Dec 2013 were retrospectively analyzed.Results There were ten cases of IPAS confirmed pathologically.Only 1 of the ten cases was diagnosed correctly and definitely with IPAS preoperatively.The other 9 cases were misdiagnosed with benign or malignant pancreatic tumors,including nonfunctional neuroendocrine neoplasms in 5 cases,pancreatic neuroendocrine cancers in 3 and pancreatic intraductal adenocarcinoma in one.All the nine misdiagnosed patients has no specific symptoms or laboratory indexes.All the IPASs located in the tail of the pancreas with the mean diameter (1.3 ±0.2) cm(0.8-2.5 cm).7 cases of IPAS show strikingly similar dynamic enhancement to the spleen on the CT scans and/or MRI.Accessory spleen around the splenic hilum was found in five cases.Conclusions Morphological study plays an important role in the diagnosis and IPAS carries parallel dynamic enhancement to the spleen on CT scans and/or MRI.IPAS should be considered as a differential diagnosis while the lesion is no more than 2.5 cm in diameter and when other accessory spleens were shown around the splenic hilum.

Objective To investigate the effects of different concentrations of sevoflurane on adhesion and expression of CD 44 in human breast cancer cell line MCF‐7 .Methods The cells were randomly divided into 4 groups :control group and 3 sevoflurane groups exposed to 1 .7% ,3 .4% and 5 .1% sevollurane for 2 ,4 and 6 h respectively .Cell adhesion rate was detected by adhesion test and the expression of CD44 mRNA and protein was determined by qRT‐PCR .Results in Treat 1 ,2 ,3 ,the adhesion rate at 2 ,4 ,6 h were lower than that before treat ,mRNA expression of CD44 reduced (P<0 .05);the adhesion rate of Treat 2 and Treat 3 at 4 and 6 h were lower than that of 2 h ,and the mRNA expression of CD44 reduced (P<0 .05);the adhesion rate of Treat 2 and Treat 3 at 6 h were lower than that of 4 h ,and the mRNA expression of CD44 reduced (P<0 .05);compared with Control group ,the adhesion rate of Treat 1 ,2 and 3 at 2 ,4 ,6 h were all lower ,mRNA expression of CD44 reduced (P<0 .05);the differences between each group were statisyicaly significant(P<0 .05) .Conclusion Sevoflurane can inhibit cell adhesion in a concentration and duration of exposure dependent manner ,and its mechanism could be related with the down regulation of CD44 expression .

ObjectiveTo investigate eutopic endometrial stromal cells' ability of proliferation,apoptosis and invasion in ( ovarian endometriosis,OEMs).Methods Culture and identify eutopic endometrial stromal cells in 32 cases of patients with OEMs and 32 cases of patients with benign teratoma.Apply methyl thiazolyl tetrazolium methods,flow cytometric and transwell methods to detect these cells' capacity of proliferation,apoptosis and invasion.ResultsEutopic endometrial stromal cells were cultured successfully and their purity was more than 90％.The zero hour's absorbance of OEMs group was similar with control group (0.127 ±0.013vs 0.129 ±0.008,P ＞ 0.05).The growth of 24 and 48 hour's absorbance of OEMs group was significantly higher than control group (24 h -0 h:0.148 ±0.020 vs 0.048 ±0.008,t =26.066;48 h -24 h:0.397 ±0.029vs 0.119 ±0.022,t =42.544,P ＜0.01 ).The apoptosis rate of eutopic endometrial stromal cells in OEMs group was (26.430 ± 3.789 )％ and ( 35.571 ± 4.485 ) ％ in control group,which reached statistical difference ( t =- 8.808,P ＜0.01 ).After the eutopic endometrial stromal cells got through the small room,the absorbance in OEMs group(0.950 ± 0.014) was significantly higher than control group (0.653 ± 0.028 ) ( t =52.947,P ＜0.01 ).ConclusionThe proliferation and invasion capacity of eutopic endometrial stromal cell in OEMs group was more powerful than control group,however the apoptosis ability of this cell in OEMs group was weaker than control group.The change of biological characteristics of eutopic endometrial stromal cells of OEMs might be involved in the occurrance and development of OEMs.

Objective To examine the expression of response gene to complement 32 (RGC-32) in renal tissue of children with IgA nephropathy (IgAN), and to explore its significance. Methods The subjects were 45 children diagnosed as IgAN by renal biopsy. The expression of RGC-32, α-smooth muscle actin (α-SMA) and transforming growth factor β1 (TGF-β1) was examined by immunohistochemistry staining. The correlation of RGC-32 expression with α-SMA,TGF-β1, degree of renal pathological lesions and clinical index in IgAN was assessed by Spearman correlation analysis. Results RGC-32 protein located in renal tubular epithelial cells in normal and IgAN renal tissues. The positive expression index of RGC-32 in nomal group, IgAN mild group, moderate group and severe group was (18.29±6.22)%, (23.90±9.65)%, (31.23±9.86)%,and (34.52±10.63)% respectively. With more severity of renal pathological lesions, the expression of RGC-32 in IgAN was enhanced. The RGC-32 expression was positively correlated with the score of glomerulus and renal interstitium in children with IgAN (r=0.385, 0.347, P＜0.05), as well as α-SMA, TGF-β1 (r=0.594, 0.521, P＜0.01), but was not correlated with Scr, urinary NAG/Cr,Alb/Cr, IgG/Cr, and α1-M/Cr (r =0.117, -0.115, -0.138, -0.176, -0.028, all P ＞0.05).Conclusions RGC-32 protein locates in renal tubular epithelial cells in normal and IgAN renal tissues. RGC-32 may participate in the course of renal tubulointerstitial lesions in children with IgAN, especially in the course of epithelial-mesenchymal transition (EMT) induced by TGF-β1.

Objective To assess the prognosis and effect on renal function of pediatric urolithiasis caused by melamine-contaminated milk powder (PUMMP) in a long-term follow-up.Methods One hundred and two of 8335 children (≤ 6-year-old) with history of consuming melamine-contaminated milk powder screened in our hospital were followed up for eighteen months after diagnosis. Urinary system ultrasonography, urinalysis, urinary microprotein profiles [microalbumin (ALBU), immunoglobulin G (IgG), and N-acetyl-β-D-glucosidase (NAG)], urinary melamine and cyanuric acid were examined in the first visit and at the end of follow-up. Results Follow-up was completed in 91 children and the stone was excreted in 82 children (90.1%).Stones less than 5 mm in diameter were most vulnerable to discharge, and stones larger than 10 mm could not be expelled without interventions. At the end of follow-up, no melamine or cyanuric acid was found in the urine samples of 74 patients. Urinalysis showed that incidences of proteinuria, microscopic hematuria and leukocyturia were 0%, 5.1% and 2.0%, which were significant different from those in the first visit (Pproteinutria=0.123, Phemnatuna=0.038 and Pleukocyhuris=0.005).Urinary microprotein profiles revealed that some children whose urinalysis was normal still presented glomerular and renal tubular injury and the abnormal rates were 8.8% and 12.1%respectively. The glomerular injury was mainly related to persistent stone, male and younger.Conclusions 90.1% of children with PUMMP passes urinary stones at the end of follow-up.Stone size is the major risk factor of discharge. No melamine or cyanuric acid is found in the urine of children. After eighteen months, glomerular and renal tubular injury is still found in some patients. Further follow-up is necessary.

Objective To investigate the clinical features,diagnosis and prognosis of solid pseudopapillary tumor of the pancreas(SPTP). Method The clinical data of SPTP cases that underwent surgical resection with a definitive histological diagnosis in Zhong Shan Hospital from 1999 to 2007 were retrospectively analyzed.Result There were 42 SPTP cases undergoing surgery in this period,including 37 females and 5 males:mean age was 37.6 years.Tumors located in the pancreatic head in 18 cases and in the body or tail in 24 cases.The most common symptom was abdominal discomfort(n=20),palpable mass (n=8).Preoperative diagnostic accuracy of MRI and CT were 93%and 31.8%,respectively.Ten cases underwent pancreaticoduodenectomy,20 cases received distal pancreatectomy,3 cases did spleen-preserving distal pancreatectomy.Tumors were simply enucleated in 8 cases.Average diameter of the tumors was 6.1 cm,34 lesions were solid,6 were solid-cystic,2 were cystic.The overall perioperative morbidity was 38.1%,10 cases developed postoperative pancreatic fistula.The mean follow-up time was 38.6 months.Except three cases that were lost to follow-up,all the other cases were alive.Four cases(2 cases had had tumor enucleation,2 had had distal pancreatectomy)developed hepatic metastases on 70、110、41、3 months after first surgery,respectively.A transarterial chemoembolization was employed in three cases,while right hemihepatectomy was performed in one case. Conclusion SPTP is an indolent tumor with low-grade biological aggressiveness.Patients had a favorable outcome after surgical treatment,but enucleation should be avoided.Patients with liver metastasis may benefit from chemoembolization and liver resection.