[Clinical Features of Infection in Newly Diagnosed Patients with Myelodysplastic Syndrome and Its Correlation with Curative Effect].

OBJECTIVE: To explore the characteristics of infection in patients with myelodysplastic syndromes (MDS), risk factors of serious infection, and their correlation with curative effect. METHODS: The clinical data of 92 newly diagnosed MDS patients with nosocomial infection from January 2016 to June 2020 in our hospital were retrospectively analyzed. RESULTS: A total of 306 courses of treatment were completed in 92 newly diagnosed MDS patients. The infection rate was the highest in the first course of treatment (84.8%, 78/92), and then decreased gradually. The top three infection sites were lung, upper respiratory tract, and gastrointestinal tract. A total of 90 strains of pathogenic bacteria were detected, of which 33.4% (30/90) were gram-negative bacilli, 23.3% (21/90) were gram-positive cocci, 23.3% (21/90) were fungi, and 20.0% (18/90) were viruses. The serious infection rate among 92 patients with MDS was 22.8% (21/92). Multivariate analysis showed that neutrophil deficiency>7 days (OR=10.875, 95%CI: 2.747-43.051, P=0.001) was an independent risk factor for serious infection in MDS patients. Compared with non-severe infection group, the total effective rate of severe infection group was lower (90.9% vs 63.6%, χ2=4.393, P<0.05). CONCLUSION: The infection rate of MDS patients is high in the first course of treatment, and the most common infection site is the lung. Gram-negative bacteria is the most common pathogen. MDS patients with neutrophil deficiency>7 days have a high risk of serious infection and poor efficacy.

LncRNA lnc-ISG20 promotes renal fibrosis in diabetic nephropathy by inducing AKT phosphorylation through miR-486-5p/NFAT5.

Long non-coding RNA (lncRNA) lnc-ISG20 has been found aberrantly up-regulated in the glomerular in the patients with diabetic nephropathy (DN). We aimed to elucidate the function and regulatory mechanism of lncRNA lnc-ISG20 on DN-induced renal fibrosis. Expression patterns of lnc-ISG20 in kidney tissues of DN patients were determined by RT-qPCR. Mouse models of DN were constructed, while MCs were cultured under normal glucose (NG)/high glucose (HG) conditions. The expression patterns of fibrosis marker proteins collagen IV, fibronectin and TGF-ß1 were measured with Western blot assay. In addition, the relationship among lnc-ISG20, miR-486-5p, NFAT5 and AKT were analysed using dual-luciferase reporter assay and RNA immunoprecipitation. The effect of lnc-ISG20 and miR-486/NFAT5/p-AKT axis on DN-associated renal fibrosis was also verified by means of rescue experiments. The expression levels of lnc-ISG20 were increased in DN patients, DN mouse kidney tissues and HG-treated MCs. Lnc-ISG20 silencing alleviated HG-induced fibrosis in MCs and delayed renal fibrosis in DN mice. Mechanistically, miR-486-5p was found to be a downstream miRNA of lnc-ISG20, while miR-486-5p inhibited the expression of NFAT5 by binding to its 3'UTR. NFAT5 overexpression aggravated HG-induced fibrosis by stimulating AKT phosphorylation. However, NFAT5 silencing reversed the promotion of in vitro and in vivo fibrosis caused by lnc-ISG20 overexpression. Our collective findings indicate that lnc-ISG20 promotes the renal fibrosis process in DN by activating AKT through the miR-486-5p/NFAT5 axis. High-expression levels of lnc-ISG20 may be a useful indicator for DN.

Exosomal microRNA-16-5p from human urine-derived stem cells ameliorates diabetic nephropathy through protection of podocyte.

Diabetic nephropathy (DN) remains one of the severe complications associated with diabetes mellitus. It is worthwhile to uncover the underlying mechanisms of clinical benefits of human urine-derived stem cells (hUSCs) in the treatment of DN. At present, the clinical benefits associated with hUSCs in the treatment of DN remains unclear. Hence, our study aims to investigate protective effect of hUSC exosome along with microRNA-16-5p (miR-16-5p) on podocytes in DN via vascular endothelial growth factor A (VEGFA). Initially, miR-16-5p was predicated to target VEGFA based on data retrieved from several bioinformatics databases. Notably, dual-luciferase report gene assay provided further verification confirming the prediction. Moreover, our results demonstrated that high glucose (HG) stimulation could inhibit miR-16-5p and promote VEGFA in human podocytes (HPDCs). miR-16-5p in hUSCs was transferred through the exosome pathway to HG-treated HPDCs. The viability and apoptosis rate of podocytes after HG treatment together with expression of the related factors were subsequently determined. The results indicated that miR-16-5p secreted by hUSCs could improve podocyte injury induced by HG. In addition, VEGA silencing could also ameliorate HG-induced podocyte injury. Finally, hUSC exosomes containing overexpressed miR-16-5p were injected into diabetic rats via tail vein, followed by qualification of miR-16-5p and observation on the changes of podocytes, which revealed that overexpressed miR-16-5p in hUSCs conferred protective effects on HPDCs in diabetic rats. Taken together, the present study revealed that overexpressed miR-16-5p in hUSC exosomes could protect HPDCs induced by HG and suppress VEGFA expression and podocytic apoptosis, providing fresh insights for novel treatment of DN.

[Retracted] MicroRNA­21 promotes the progression of peritoneal fibrosis through the activation of the TGF­ß/Smad signaling pathway: An in vitro and in vivo study.

Int J Mol Med 41: [Related article:] 1030­1038, 2018; DOI: 10.3892/ijmm.2017.3268. An interested reader drew to our attention the fact that the western blots featured in Fig. 2B in the above article contained duplicated data: The data shown for the TGF­ß and vimentin protein bands were apparently identical; furthermore, there was a strong likelihood that the protein bands featured for the ZO­1 and SMAD­3 experiments were also the same, but flipped horizontally relative to the other. Following an investigation, the Journal was able to confirm that this duplication of the research data had probably occurred. On those grounds, the Editor of International Journal of Molecular Medicine has decided that the above paper should be retracted. We were unable to make contact with the authors of the article published in International Journal of Molecular Medicine, despite every effort to do so. The Editor deeply regrets any inconvenience that this retraction has caused to the the readership of the Journal.

MicroRNA-21 promotes the progression of peritoneal fibrosis through the activation of the TGF-ß/Smad signaling pathway: An in vitro and in vivo study.

The present study aimed to explore the roles of microRNA-21 (miR­21) and the transforming growth factor-ß (TGF-ß)/Smad signaling pathway in the development of peritoneal fibrosis (PF). First, dialysis effluents from 30 patients with PF were collected, and after the establishment of a mouse model of PF, hematoxylin and eosin (H&E) and Masson's staining were used to observe peritoneal tissues, inflammatory cells and blood vessels. High glucose was used to stimulate human peritoneal mesothelial cell lines and these stimulated cells were then transfected with miR­21 inhibitor. Immunofluorescence microscopy was applied for the observation of the transfected cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR­21, and RT-qPCR and western blot analysis were used to detect the mRNA and protein expression of Zonula occludens-1 (ZO-1), TGF-ß, Smad, vimentin and connective-tissue growth factor (CTGF). The mRNA and protein expression levels TGF-ß, Smad-3, vimentin and CTGF were elevated, while ZO-1 mRNA and protein expression was decreased with the prolonged duration of dialysis treatment in the patients with PF. The experiments using the mouse model of PF revealed that the peritoneal connective tissue was thickened, while the numbers of inflammatory cells and blood vessels were increased. The expression levels of miR­21, and the mRNA and protein expression levels of TGF-ß, Smad-3, vimentin and CTGF were increased over time, whereas the mRNA and protein expression levels ZO-1 constantly decreased in the mice in the experimental group. Moreoever, the expression of miR­21 positively correlated with the expression levels of TGF-ß, Smad-3, vimentin and CTGF, while it negatively correlated with the expression of ZO-1. The results of H&E and Masson's staining revealed that miR­21 expression was associated with the degree of PF. These findings thus indicate that miR­21 promotes the progression of PF through the activation of the TGF-ß/Smad signaling pathway.

Whole exome sequencing identifies mutation of EDNRA involved in ACTH-independent macronodular adrenal hyperplasia.

ACTH independent macronodular adrenal hyperplasia (AIMAH) is a rare disorder characterized by bilateral macronodular hyperplasia of the adrenal glands and increased cortisol production with subclinical or overt Cushing's syndrome. Although the family clustering of AIMAH is infrequent, we have tried our best to find such a familial affected pedigree with complete clinical information and successfully collect adrenalectomy tissue samples from two members of this family. Using whole exome sequencing and several variant prioritization strategies based on disease network analysis, we identified Endothelin receptor type A (EDNRA) Ser420Thr mutation as a causative mutation of AIMAH. EDNRA is a member of G protein coupled receptor family and is involved in cardiovascular or polycystic kidney disease. Our findings indicate that the mutation of EDNRA at S420T site should be regard as a potential AIMAH causative variation in familial and sporadic affected patients.

[Frequency and linkage disequilibrium of specific HLA-DR and HLA-DQ genes in Chinese Han population].

This study was aimed to investigate the distribution feature of HLA-DR/DQ gene linkage disequilibrium in Chinese Han population and to improve the accuracy of HLA matching results. Genotyping of HLA-DR and HLA-DQ gene locus was performed using PCR-SSP typing in Chinese Han population receiving kidney transplantation. The results showed that there were 29 new linkage combinations in 1799 patients, in which DR13-DQ5, DR11-DQ8 and DR8-DQ8 were discovered for 11, 8 and 7 times respectively while DR9-DQ8, DR12-DQ6 and DR14-DQ4 were both discovered for 6 times. The linkage disequilibrium parameters of these haplotypes were negative, showing that these linkages were uncommon. It is concluded that this study not only enriches the classical HLA-DR/DQ linkage combinations, but also indicates the national relevance of combination distribution, and it has great importance in improving the accuracy of HLA matching experiments and reducing unnecessary repeated work.

Anatomical variation of the posterior lumbar tributaries of the left renal vein in retroperitoneoscopic left living donor nephrectomy.

OBJECTIVES: To increase awareness of the anatomical variation of the posterior lumbar tributaries of the left renal vein in retroperitoneoscopic left living donor nephrectomy. METHODS: A total of 61 cases of retroperitoneoscopic left living donor nephrectomy were carried out from March 2008 to June 2010. The anatomical variations of the posterior lumbar tributaries of the left renal vein in these patients were noted. RESULTS: According to the variation of posterior lumbar tributaries, there were seven types in total, including five main types (accounts for 95.1%, 58/61 cases) and the type of reno-hemi-azygo-lumbar trunk (AZV; accounts for 16.4%, 10/61 cases). According to the number of posterior lumbar tributaries, no lumbar vein covers accounted for 16.4% (10/61 cases), one lumbar vein accounted for 47.5% (29/61 cases), two lumbar veins accounted for 32.8% (20/61 cases) and three lumbar veins accounted 3.3% (2/61 cases). According to the operation time during the process of managing posterior lumbar veins, it was type 4 (AZV) on which the surgeon spent the most time (P<0.05), and type 5 (no lumbar vein) on which the surgeon spent the least time (P<0.05). CONCLUSIONS: This is the first report of the anatomical variation of the posterior lumbar tributaries of the left renal vein in retroperitoneal laparoscopic left living donor nephrectomy. Detailed knowledge of these anatomical variations will undoubtedly help surgeons to avoid the potential risk of vein damage during nephrectomy and to obtain a longer renal artery for the following renal transplantation.

[In vitro expansion of T cells stimulated by combination of IL-2, IL-7 and IL-15].

The aim of this study was to compare cell proliferation and function of the T cells acquired under various culture conditions for establishing a simple, safe and efficient cell expansion protocol in vitro. The peripheral blood mononuclear cells (PBMNC) were isolated and stimulated with autologous dendritic cells (DC) and EBV-transformed B lymphoblastoid cell line (BLCL) weekly. The cell proliferation test, flow cytometry with PI and Annexin V double staining, Cr release test and ELISPOT test were used to detect the cell expansion level, frequency of IFN-γ producing T cells, killing activity of antigen-specific T cells, cell apoptotic status and cell differentiation potential, respectively. The results indicated that use of IL-2 combined with IL-7 and IL-15 resulted in the highest cell expansion comparing to the use of IL-2 alone and the use of CD3/28 Microbeads. Also the cells obtained under cultivating with IL-2, IL-7 and IL-15 together showed high frequency of IFN-γ producing cells, strong killing activity, high viability and high differentiation potential with large portion of CD3(+)CD8(+) population among the T cells. It is concluded that a protocol is established in which the use of IL-2 combined with IL-7 and IL-15 induces the biggest cell expansion, expanded cells show high viability, strong differentiation potential, high frequency of IFN-γ producing cells and strong killing activity.

[Retroperitoneal laparoscopic living donor nephrectomy: report of 58 cases].

OBJECTIVE: To review a single-institution experience with retroperitoneal laparoscopic living donor nephrectomy (RLDN). METHODS: Fifty-eight donors underwent RLDN at our institution (including 32 male and 26 female donors aged 20-61 years, mean 42 years). Left nephrectomy was performed in 56 cases. The first 35 patients underwent total RLDN, and the latter 23 received modified RLDN. RESULTS: RLDN was performed successfully in all the patients without conversion to open surgery. The mean surgical time was 93 min (range 70-130 min), and the mean blood loss was 20 ml (range 10-50 ml), with a mean warm ischemia time of 2.8 min (1.3-6 min). Retroperitoneal hematoma occurred postoperatively in one case. The mean hospital stay of the donors was 6.4 days (5-10 days). Two recipients showed delayed graft function, and one graft was lost because of acute rejection. The other recipients had normal renal function in two weeks except for 3 having normal renal function in 4 weeks. CONCLUSION: RLDN is a safe procedure with minimal invasiveness, and the modified RLDN lowers the learning curve of the surgery.

[Gene diagnosis of facioscapulohumeral muscular dystrophy].

OBJECTIVE: To observe the characteristics of changes of p13E-11 labelled 4q35 EcoRI fragments and to make a gene diagnosis of facioscapulohumeral muscular dystrophy(FSHD). METHODS: Genomic DNA was extracted and was digested by EcoR I /Bln I. After pulsed field gel electrophoresis, it was hybridized with probe p13E-11 by Southern blot. The illness was diagnosed as FSHD when the 4q35 EcoRI fragment was smaller than 38 kb. RESULTS: In 26 cases of FSHD, the fragments of 20 cases were smaller than 38 kb. The positive rate was 76.92%. In 12 cases of FSHD family members, the fragments of 2 cases were smaller than 38 kb. All fragments of the 21 controls were greater than 38 kb. CONCLUSION: It was rather good to use <38 kb as a standard for diagnosis of FSHD. The positive rate of FSHD was similar to that from the references.