Asiatic acid improves insulin secretion of β cells in type 2 diabetes through TNF- α/Mfn2 pathway / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effects and molecular mechanisms of asiatic acid on β-cell function in type 2 diabetes mellitus (T2DM).@*METHODS@#The T2DM model was established by high fat diet and streptozotocin injection in ICR mice, and the effects of asiatic acid on glucose regulation were investigated in model mice. The islets were isolated from palmitic acid-treated diabetic mice. ELISA was used to detect the glucose-stimulated insulin secretion, tumor necrosis factor (TNF)-α and interleukin (IL)-6. ATP assay was applied to measure ATP production, and Western blotting was used to detect protein expression of mature β cell marker urocortin (Ucn) 3 and mitofusin (Mfn) 2. The regulatory effects of asiatic acid on glucose-stimulated insulin secretion (GSIS) and Ucn3 expression were also investigated after siRNA interference with Mfn2 or treatment with TNF-α.@*RESULTS@#Asiatic acid with the dose of 25 mg·kg－1·d－1 had the best glycemic control in T2DM mice and improved the homeostasis model assessment β index. Asiatic acid increased the expression of Mfn2 and Ucn3 protein and improved the GSIS function of diabetic β cells in vitro and in vivo (both P<0.05). Moreover, it improved the ATP production of islets of T2DM mice in vitro (P<0.05). Interfering Mfn2 with siRNA blocked the up-regulation of Ucn3 and GSIS induced by asiatic acid. Asiatic acid inhibited islet TNF-α content and increased Mfn2 and Ucn3 protein expression inhibited by TNF-α.@*CONCLUSIONS@#Asiatic acid improves β cell insulin secretion function in T2DM mice by maintaining the β cell maturity, which may be related to the TNF-α/Mfn2 pathway.

Clinical features and genetic analysis of two children with Williams-Beuren syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical and genetic characteristics of two children with Williams-Beuren syndrome (WBS).@*METHODS@#Two children who had presented at the Department of Pediatrics, General Hospital of Ningxia Medical University respectively on January 26 and March 18, 2021 were selected as the study subjects. Clinical data and results of genetic testing of the two patients were analyzed.@*RESULTS@#Both children had featured developmental delay, characteristic facies and cardiovascular malformation. Child 1 also had subclinical hypothyroidism, whilst child 2 had occurrence of epilepsy. Genetic testing revealed that child 1 has harbored a 1.54 Mb deletion in the 7q11.23 region, whilst child 2 has a 1.53 Mb deletion in the same region, in addition with a c.158G>A variant of the ATP1A1 gene and a c.12181A>G variant of the KMT2C gene. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.158G>A and c.12181A>G variants were rated as variants of unknown significance (PM1+PM2_Supporting+PP2+PP3；PM2_Supporting).@*CONCLUSION@#Both children had characteristic features of WBS, for which deletions of the 7q11.23 region may be accountable. For children manifesting developmental delay, facial dysmorphism and cardiovascular malformations, the diagnosis of WBS should be suspected, and genetic testing should be recommended to confirm the diagnosis.

Heterogeneity of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation based on latent profile analysis / 器官移植

Objective To explore heterogeneous subtypes of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation and the characteristics differences of different types of children after liver transplantation. Methods Seven hundred and forty-one children who underwent living-related liver transplantation were enrolled. The self-designed general information questionnaire, Chinese version of 5-Item World Health Organization Well-Being Index (WHO-5) and the parent-report version of the Strengths and Difficulties Questionnaire (SDQ) were filled out by their guardians. The scores of five dimensions of SDQ were used as the manifest variables of the model. The classification model of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation was constructed by latent profile analysis. The latent categories of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation were analyzed. The influencing factors of latent categories were analyzed by univariate analysis and logistic regression model. Results There were three latent categories of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation, including peer communication problem group (n=302), psychological and behavioral adaptation group (n=145) and psychological and behavioral adjustment difficulty group (n=294). The first two groups were merged into the psychological and behavioral health group (n=447), which had significant differences in the five dimensions and the total score of difficulties of SDQ compared with the psychological and behavioral adjustment difficulty group (n=294) (all P<0.001). Logistic regression analysis showed that age≤5 years old, primary disease of non-cholestatic liver disease, stem family were the risk factors for psychological and behavioral adjustment difficulties in pediatric recipients after liver transplantation. Female gender, high education levels of parents and high WHO-5 score of guardians were the protective factors for psychological and behavioral adjustment difficulties in pediatric recipients after liver transplantation (all P<0.05). Conclusions The psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation are heterogeneous. Medical staff should pay extensive attention to different characteristics of pediatric recipients after liver transplantation with different psychological and behavioral adaptation categories and adopt targeted screening and intervention strategies, aiming to improve psychological and behavioral adaptation outcomes of pediatric recipients after liver transplantation.

Mitofusin2 Promotes ß Cell Maturation from Mouse Embryonic Stem Cells via Sirt3/Idh2 Activation.

ß cell dysfunction is the leading cause of diabetes. Adult ß cells have matured glucose-stimulated insulin secretion (GSIS), whereas fetal and neonatal ß cells are insensitive to glucose and are functionally immature. However, how ß cells mature and acquire robust GSIS is not fully understood. Here, we explored the potential regulatory proteins of ß cell maturation process and the capacity for GSIS. Combined with the data from public databases, we found that the gene expression of Mitofusin2 (Mfn2) showed an increasing trend from mouse neonatal ß cells to mature ß cells. Moreover, its protein expression increased during mouse embryonic pancreas development and ß cell differentiation from mouse embryonic stem cells. Knocking down Mfn2 reduced Urocortin3 (Ucn3) expression, GSIS, and ATP production in induced ß cells, while overexpressing it had the opposite effect. However, neither Mfn2 knockdown nor overexpression affected the differentiation rate of insulin-positive cells. In immature and mature ß cells, Mfn2 and its correlated genes were enriched in tricarboxylic acid (TCA) cycle-related pathways. The expressions of Sirtuin 3 (Sirt3) and isocitrate dehydrogenase 2 (NADP+) and mitochondrial (Idh2) were Mfn2-regulated during ß cell differentiation. Inhibiting Idh2 or Sirt3 reduced cellular ATP content and insulin secretion levels that increased by Mfn2 overexpression. Thus, Mfn2 modulated the induced ß cell GSIS by influencing the TCA cycle through Sirt3/Idh2 activation. We demonstrated that Mfn2 promoted embryonic stem cell-derived ß cell maturation via the Sirt3/Idh2 pathway, providing new insights into ß cell development. Our data contribute to understanding diabetes pathogenesis and offer potential new targets for ß cell regeneration therapies.

A systematic review of enteral nutrition versus parenteral nutrition on patient outcomes after liver transplantation / 中华器官移植杂志

Objective:To systematically evaluate the therapeutic efficacy of enteral nutrition(EN)versus parenteral nutrition(PN)in patients after liver transplantation(LT).Methods:The databases of Cochrane Library, MEDLINE, EmBase, EBSCO, Sinomed, CBM, CNKI and Wanfang Data were systematically searched up to August 2021 for collecting randomized controlled trials(RCT)of comparing EN and PN in patients after LT.In strict accordance with the inclusion and exclusion criteria, two independent researchers carried out literature screening, data extracting and literature methodological quality evaluations.Then RevMan5.3 software was utilized for quantitative synthesis and statistical processing.Results:A total of 13 RCTs involving 571 grade B patients were included.Meta-analysis indicated that, as compared with PN, serum albumin of EN spiked[WMD=2.5(1.33, 3.68), WMD>0, P<0.000 1], prealbumin rose[WMD=59.78(32.22, 87.35), WMD>0, P<0.000 1], total bilirubin dropped[WMD=-27.81(-41.88, -13.74), WMD<0, P=0.000 1]and infection incidence was significantly lower[ OR=0.25(0.16, 0.39), OR<1, P<0.000 01]. Conclusions:After LT, as compared with PN, EN can boost the nutritional level, promote the recovery of liver function and lower the incidence of postoperative infections.It offers some clinical values.

Nursing experience in the process of diagnosing and treating post-transplantation lymphoproliferative disorder / 中国实用护理杂志

Objective@#To explore the key points of nursing in the process of diagnosing and treating the common complication--- post-transplantation lymphoproliferative disorder (PTLD) after pediatric liver transplantation.@*Methods@#The clinical data of 21 children with PTLD after liver transplantation. The key point of nursing in the process of prevention, onset, diagnosis and treatment was summarized.@*Results@#With the individual and combined treatments 18 patients gained remission except for 1 patient died because of perforation and sequent intractable haemorrhage and 2 patient died for the development of lymphoma. In terms of nursing, during the prevention stage, it is critical to insist the regular follow-up to make early diagnosis. More efforts should be made to obtain the children′s cooperation to complete the examination. During the infusion of rituximab, strengthened monitoring is necessary and more attention should be paid to the special nursing when other complications such as intractable haemorrhage, intestinal obstruction or perforation occurs.@*Conclusions@#The nursing care of PTLD is complicated. Individual and careful nursing regimens should be made based on the specific situations in different stages.

Nursing experience in the process of diagnosing and treating post-transplantation lymphoproliferative disorder / 中国实用护理杂志

Objective To explore the key points of nursing in the process of diagnosing and treating the common complication--- post-transplantation lymphoproliferative disorder (PTLD) after pediatric liver transplantation. Methods The clinical data of 21 children with PTLD after liver transplantation. The key point of nursing in the process of prevention, onset, diagnosis and treatment was summarized. Results With the individual and combined treatments 18 patients gained remission except for 1 patient died because of perforation and sequent intractable haemorrhage and 2 patient died for the development of lymphoma. In terms of nursing, during the prevention stage, it is critical to insist the regular follow-up to make early diagnosis. More efforts should be made to obtain the children′s cooperation to complete the examination. During the infusion of rituximab, strengthened monitoring is necessary and more attention should be paid to the special nursing when other complications such as intractable haemorrhage, intestinal obstruction or perforation occurs. Conclusions The nursing care of PTLD is complicated. Individual and careful nursing regimens should be made based on the specific situations in different stages.

MicroRNA expression analysis of mammospheres cultured from human breast cancers.

PURPOSE: There is accumulating evidence suggests that tumors are initiated and maintained by a small fraction of tumor-initiating cells (TICs). TICs can be enriched by mammospheres culturing without surface markers. MicroRNAs participated in many important processes of life including regulating tumorigenicity of TICs. However, roles of miRNAs in TICs of breast cancer are still unknown. METHODS: We compared mammospheres formation of four breast cancer cell lines, cultured mammospheres from breast cancers specimens of three patients and compared microRNAs profiling of mammospheres cultured from breast cancer specimens with differentiated progenies. RESULTS: Three of the four breast cancer cell lines showed the ability of mammospheres formation. The proportions of CD24(-)cells in mammospheres were increased significantly in the three cell lines. The expression of genes associated with stem cells and chemoresistance increased significantly after mammospheres formation. Breast cancer cells isolated from patients' specimens survived in nonadherent culture conditions generated mammospheres with ability of self-renewal and bilineage potential. MicroRNA expression profiling of mammospheres compared with differentiated progenies isolated and propagated from the three patients identified 17 microRNAs. And the target genes of these miRNAs are involved in several key signaling pathways. CONCLUSIONS: The results suggested that mammospheres were enriched in TICs and proved a valuable model for studies of breast cancer TICs in vitro, microRNAs played critical roles in maintenance of stemness properties of mammospheres and provided novel insights into breast cancer therapy.

Inhibition effects of monoclonal antibody C225 on breast cancer stem cells / 上海交通大学学报（医学版）

Objective To explore the inhibition effects of epidermal growth factor receptor (EGFR) antagonist monoclonal antibody cetuximab (C225) on breast cancer stem cells in human breast cancer cell line MCF-7.Methods The effects of C225 on the proliferation of breast cancer cell line MCF-7 were detected by MTT assay. MCF-7 cells were cultured to generate primary mammospheres, and were divided into control group, C225 group, epidermal growth factor (EGF) group and EGF + C225 group according to whether or not the culture media contained exogenous EGF and C225.Thirteen days after culture, the volume and number of mammospheres of these four groups were observed, and mammosphere-forming efficiency ( MFE) was calculated. The percentages of CD44~+ CD24~- cells in mammospheres of these four groups and in routinely cultured MCF-7 cells were determined by flow cytometry. Results The inhibition rate on MCF-7 cells increased with the concentration of C225.Compared with control group, the volume of mammospheres in C225 group significantly decreased, and MFE and percentages of CD44~+ CD24~- cells in mammospheres significantly decreased [(0.61 ±0.04)% vs (1.44±0.09)%, P<0.01; (3.50±0.29)% vs (9.07 ±0.52) % , P<0.01]. Compared with EGF group, the volume of mammospheres in EGF + C225 group significantly decreased, and MFE and percentages of CD44~+ CD24~- cells in mammospheres significantly decreased [ (0.68 ± 0.04) % vs (1.61 ± 0.05) % , P < 0.01; (4.00 ± 0.58) % vs (10.47 ± 0.79) % , P < 0.01]. The percentage of CD44~+ CD24~- cells in routinely cultured MCF-7 cells was (2.03 ±0.15) % , and was significantly different from those in EGF group and control group (P < 0.01). There was no significant difference in volume of mammospheres, MFE and percentage of CD44~+ CD24~- cells in mammospheres between EGF group and control group (P >0.05). Conclusion C225 has significant inhibition effects on CD44~+ CD24~- cells in MCF cells.

Effects of different dialysates on apoptosis and expression of PKCδ of U937 cell line / 上海交通大学学报（医学版）

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group , normal control group and fluid A+powder B+rottlerin group (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P＞0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

Influence factors on the formation of mammospheres from breast cancer stem cells / 北京大学学报(医学版)

Objective:To investigate the influential factors on the formation of mammospheres from MCF-7,MDA-MB-231 and primary breast cancer epithelial cells,and to detect the proportion of CD44+/CD24-/low among them.Methods:MCF-7,MDA-MB-231 and primary breast cancer epithelial cells were cultured in suspension to generate primary mammospheres.After 7 days of cultivation,the proportion of cancer stem cells was measured in cells derived from mammosphere cells or monolayer culture cells by flow cytometry.Results:MCF-7 had the highest efficiency of mammosphere formation(2.1%?0.3%),while MDA-MB-231 cells hardly had any mammosphere formation,and no mammosphere formation was observed in breast cancer epithelial cells.MCF-7 tended to adhere under stem cell culture conditions without B27.Flow cytometry analysis indicated that,compared with 2.0%?0.1% for the MCF-7 monolayer culture cells,11.8%?0.3% and 8.2%?0.8% of the MCF-7 mammosphere cells from MCF-7 under different conditions were CD44+/CD24-/low(P