Objective To investigate the role and related mechanism of the soluble guanylate cyclase(sGC)-cGMP-protein kinase G(PKG)signaling pathway in the amelioration of isoproterenol(ISO)-induced cardiac hypertrophy in mice by aqueous extract of Curcuma kwangsiensis root tubers(GYJS).Methods 72 KM male mice were divided randomly into 6 groups:normal,model,propranolol control(40 mg/kg),and GYJS low-(1 g/kg),medium-(2 g/kg),and high-dose(4 g/kg)groups.Mice in the experimental groups were injected subcutaneously with ISO 10 mg/kg on days 1～3 and ISO 5 mg/kg on days 4～14 to establish a mouse cardiac hypertrophy model.4h after each subcutaneous injection of ISO,the mice in each group were administered the corresponding drugs orally for a dosing cycle of 14 days.The hearts were then removed and whole heart and left ventricle weight were measured.Myocardial tissue pathology was observed using hematoxylin and eosin and Masson staining,and sGC subunit beta-1(GUCY1B3)and transforming growth factor(TGF-β1)were detected by immunohistochemistry.Serum lactate dehydrogenase(LDH),creatine kinase(CK),and Nitric Oxide(NO),and myocardial superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were measured using respective kits.Serum cGMP was detected by enzyme-linked immunosorbent assay and quantitative reverse transcription PCR(RT-qPCR),and atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),GUCY1B3,PKG Ⅰ,and phosphodiesterase(PDE)5A mRNA expression levels were also determined by RT-qPCR.Results Compared with the model group,whole heart and left ventricle weights were significantly reduced in mice treated with propranolol or GYJS(P＜0.001 or P＜0.0001)and myocardial hypertrophy and myocardial fibrosis were significantly reversed.All the treatments significantly elevated myocardial expression of GUCY1B3(P＜0.05 or P＜0.01)and significantly reduced expression of TGF-β1(P＜0.05 or P＜0.01).The myocardial damage markers LDH and CK were significantly reduced(P＜0.05 or P＜0.01)while NO and cGMP were significantly elevated(P＜0.05 or P＜0.01),the myocardial oxidative stress indicator MDA was significantly reduced(P＜0.05 or P＜0.01)and SOD activity was significantly increased(P＜0.05 or P＜0.01).mRNA levels of the myocardial hypertrophy markers ANP,BNP,and PDE5A were significantly reduced(P＜0.05,P＜0.01,or P＜0.001)and the mRNA levels of GUCY1B3 and PKG Ⅰ were significantly increased(P＜0.01 or P＜0.001).Conclusions GYJS may improve cardiac hypertrophy by modulating the sGC-cGMP-PKG signaling pathway.

Multiple endocrine neoplasia-IIb (MEN-IIb) is a rare hereditary autosomal dominant syndrome caused by mutations in the RET proto-oncogene. It's characterized by medullary thyroid carcinoma (MTC), pheochromocytoma (PHEO), mucosal neuromas, and Marfanoid habitus. Because of the rarity of MEN-IIb and finiteness of clinical cognition, the majority of the patients suffer a delayed diagnosis. A MEN-IIb patient with the lingual mucosal neuromas since childhood was admitted in the Third Xiangya Hospital of Central South University in November, 2018. He had surgical history of mitral valve prolapse and spinal deformity. He was diagnosed with MTC and PHEO at the age of 22 and 28, respectively, and received surgical treatments. Sequencing of RET gene revealed a de novo heterozygous p.M918T mutation in the patient. Being aware of the unique clinical phenotype and screening of RET gene mutation may lead to the early diagnosis and better long-term outcome for MEN-IIb.
Adrenal Gland Neoplasms ; Child ; Genes ; Humans ; Male ; Multiple Endocrine Neoplasia ; Multiple Endocrine Neoplasia Type 2a/genetics* ; Multiple Endocrine Neoplasia Type 2b/genetics* ; Mutation ; Proto-Oncogene Proteins c-ret/genetics* ; Thyroid Neoplasms/genetics*

Objective: Autoimmune polyendocrine syndrome type Ⅰ(APS-Ⅰ) is caused by mutations in the autoimmune regulator gene (AIRE) gene. In this study, phenotype and AIRE gene analysis were performed in two patients with APS-Ⅰ. Methods: Peripheral blood samples were collected from two patients with APS-Ⅰ and their families. All exons of the AIRE gene and adjacent exon-intron sequences were amplified by PCR and subsequently sequenced. The silico analysis was performed to predict the possible impact of the mutations on the function of the AIRE protein. At the same time, 100 healthy controls were selected to confirm the mutation. Results: Case 1 was a 31-year-old female who exhibited chronic mucocutaneous candidiasis, hypoparathyroidism, Addison′s disease, Hashimoto′s thyroiditis, and premature ovarian failure. A homozygous c. 483_484insC mutation in exon 4 of AIRE gene was identified in this patient. Her parents, siblings and son were heterozygous for this mutation, which is consistent with the autosomal recessive inheritance pattern. Case 2 was a 34-year-old male who had mucocutaneous candidiasis, Addison′s disease, primary hypoparathyroidism, and Hashimoto′s thyroiditis. A compound heterozygous AIRE mutation (c.179A>G/C.463+ 2T>C) were identified in this patient. His father was heterozygous for c. 179A>G mutation, and his mother was heterozygous for C. 463+ 2T>C, which is consistent with autosomal recessive inheritance mode. The c. 483_484insC and c. 463+ 2T>C have been reported to be pathogenic. The c. 179A>G mutation was predicted pathogenic by SIFT and PolyPhen2 software, which was not detected in 100 healthy controls. It has not been reported in the HGDM database and is a novel mutation. Conclusion We identified a novel AIRE gene mutation (c.179A>G), which contributed to further understanding of the pathogenesis of APS-Ⅰ. The clinical variation and rarity of APS-Ⅰ makes the syndrome hard to recognize. Early recognition of symptoms and screening for AIRE mutation in patients with APS-Ⅰ has important clinical implications for the diagnosis and treatment.

Two patients with primary hypertrophic osteoarthropathy (PHO) and their available healthy family members were studied.All exons of the SLCO2A1 and HPGD gene and adjacent exonintron sequences were amplified by PCR and subsequently sequenced.To assess the damaging effects of missense mutations in silico,the online database,PolyPhen-2 and SIFT were used.We identified two homozygous mutations in SLCO2A1 gene:one was c.1106G＞A (p.G369D) in exon 9,the other was c.611C＞T (p.S204L) in exon 4.No HPGD mutation was found in the affected individuals.The two mutation were predicted in silico by the bioinformatic tools.Our study further supports the role of mutations in the SLCO2A1 gene in the pathogenesis of PHO.Identification of the genotype in PHO may not only help the clinical diagnosis of PHO but also help the interpretation of genetic information for prenatal diagnosis and genetic counseling.

Objective: To identify the differentially expressed miRNAs in the exosomes secreted from γ-ray irradiated cells and provide new clues in disclosing the mechanisms of radiation-induced bystander effects. Methods: The human bronchial epithelial cells (BEP2D) were irradiated with ⁶⁰Co γ-rays, and the exosomes were collected by ultracentrifugation from the culture medium of 2 Gy-irradiated cells and non-irradiated control. The exosomes were identified by an electron microscopy. The miRNA microarray technique was used to analyze the miRNA expression profiles in the exosomes. qRT-PCR was used to verify the miRNAs expression. The functional pathways of miRNAs targeting genes were predicted by informatic analysis using the databases of TargetScan, miRanda, GO and KEGG. Results: Sixteen miRNA with significantly increased expression (P<0.05) were identified in the exosomes of BEP2D cells at 4 h post-2 Gy irradiation as compared with the non-irradiated control cells, among which miR-100-5p, miR-1246, miR-29b-3p, and miR-7-5p were further confirmed to be unregulated by qRT-PCR assay (P<0.05). Meanwhile, the expression changes of above-mentioned four miRNAs were also investigated in the irradiated cells. The data indicated the expression was significantly increased at 2 h post-2 Gy irradiation for the miR-100-5p, miR-1246, miR-29b-3p in addition to miR-7-5p. However, all these four miRNAs were downregulated in the cells at 4 h post-irradiation and then gradually recovered. Bioinformatics analysis showed that the targeted genes of these differentially expressed miRNAs might participate in the biological processes and signal pathways of cell adhesion, mTOR signal pathway, chromatin modification, HR and NHEJ pathways of DNA repair and so on. Conclusions Radiation-inducible miRNAs have been identified in the exosomes from the irradiated BEP2D cells. The target genes of these miRNAs play roles in a series of important biological processes and functional pathways, which provides new clues in elucidating the mechanisms of radiation-induced bystander effects.

Neurofibromatosis Type 1 (NF1) is an autosomal dominant genetic disorder caused by NF1 gene mutations. Café au lait spots, neurofibromatosis, Lisch nodules, axillary freckling, dermal neurofibromas and skeletal dysplasia are the most common manifestations for this disease. A 11-year-old boy visited Third Xiangya Hospital, Central South University due to growth-retardation. He was eventually diagnosed as NF1 with growth hormone deficiency. A novel heterozygous splicing mutation c.6579+2 T>C (IVS 34+2 T>C) of NF1 gene was identified in the patient and his mother. Considering NF1 may present with short stature due to growth hormone deficiency, all children with short stature combined with café au lait spots should be screened for NF1, which may assist the clinical diagnosis and the genetic counseling.
Cafe-au-Lait Spots ; diagnosis ; genetics ; Child ; Genes, Neurofibromatosis 1 ; Growth Hormone ; deficiency ; Humans ; Male ; Mutation ; Neurofibromatosis 1 ; blood ; diagnosis

Objective To study the changes in miRNAs expression in the exosomes of human umbilical vein endothelial cells(HUVECs) after 60 Co γ-rays expose using microRNA(miRNA) chips and bioinformatics techniques so as to provide new clues to the mechanism of radiation-induced vascular tissue injury and its bystander effects.Methods HUVECs exosomes were collected in the control and 4 Gy irradiated cells by ultra-high-speed centrifugation,and further confirmed using transmission electron microscopy (TEM) and Western blotting of exosomes biomarkers.miRNA microarray was used to analyze miRNA expression profiles of exosomes and cells.Also,real-time quantitative PCR(qRT-PCR) was used to verify differentially expressed miRNAs,and the miRDB and TargetScan were performed to predict the target genes of the differentially expressed miRNAs.Bioinformatics analysis was performed using DAVID,KEGG and other online tools.Results Compared with the control exosomes from non-irradiated HUVECs,miRNA microarray analysis revealed that 5 up-regulated,and 13 down-regulated miRNAs were identified in the exosomes from HUVECs at 0.5 h after 4 Gy-irradiation,and 16 up-regulated and 5 down-regulated miRNAs at 2 h after 4 Gy-irradiation.Moreover,38 and 85 miRNAs were differentially expressed respectively in the HUVECs at 0.5 h and 2 h after radiation.The difference was statistically significant(P<0.01).The results of bioinformatics showed that these miRNAs might exert the radiation-induced bystander effect (RIBE) by regulating MAPK signal pathways,RAS and PI3K-Akt signal pathways.Conclusion The ionizing radiation injury significantly alters the components and expression levels of exosomal miRNAs,which play important roles in regulating the signal pathways in response to radiation.

Purpose To explore the predictive value of myocardial perfusion in assessing myocardial systolic function recovery after primary percutaneous coronary intervention (PPCI),in order to improve poor prognosis by early detection of myocardial no-reflow.Materials and Methods Forty nine patients with acute myocardial infarction (AMI) who had received PPCI were chosen as subjects.All the patients underwent two-dimensional strain (2DS) images and resting real-time myocardial contrast echocardiography (MCE) within one week after surgery,and 2DS measurement was repeated after three months.2DS imaging was used to acquire longitudinal peak systolic strain (LPSS) at all myocardial segments.Based on the graphs of LPSS,left ventricular myocardium was divided into normal contractile function myocardium (red) and impaired contractile function myocardium (light red,blue).According to the myocardial perfusion scores (MPS) qualitatively assessed by MCE visual interpretation,the myocardia with impaired systolic function were categorized into three groups with different perfusion level.The changes of LPSS within one week and three months after surgery (△ LPSS) among the three groups were analyzed.The correlation between MPS and LPSS within one week and three months after PPCI was also analyzed respectively.Results The △ LPSS increased significantly among the three groups with the improvement of myocardial perfusion level [(-5.78±6.23)％ vs.(-4.37±6.60)％ vs.(-1.21 ±4.77)％,all P＜0.05].The MPS measured one week after PPCI was both positively correlated with the LPSS detected within one week after surgery and that after three months (r=0.47,0.58,P＜0.001).The consistence of myocardial perfusion scores given by two evaluators was good (Kappa=0.785,P＜0.05).Conclusion The level of myocardial perfusion after PPCI in patients with AMI is closely related to regional myocardial systolic function,and the improvement of myocardial perfusion can forecast the recovery of regional systolic function.

Objective To evaluated the value of myocardial perfusion before delayed percutaneous coronary intervention (PCI) for predicting the recovery of systolic function of patients with acute myocardial infarction (AMI).Methods A total of 64 patients with AMI receiving delayed PCI treatment in the First People's Hospital of Foshan from January 2014 to June 2015 were selected.One day prior to delayed PCI,all of the patients underwent two dimensional strain to measure the longitudinal peak systolic strain (LPSS) of each left ventricular segment and the global longitudinal strain (GLS) of the left ventricle.The myocardial perfusion score (MPS) and the perfusion score index (PSI) were measured by myocardial contrast echocardiography (MCE).Left ventricular myocardial perfusions were classified as good,reduced,or absent.The two dimensional strain measurements were again conducted at 6 months after the delayed PCI to assess LPSS and GLS.The change of GLS and LPSS between one day prior to delayed PCI and six months after delayed PCI was assessed by paired t-test.The differences of LPSS among good,reduced,or absent myocardial perfusion groups were analyzed by one-way ANOVA.LSD-t test was used to compare in pairs of groups that had different values.The correlations between PSI and GLS,MPS and LPSS were assessed by Spearman's rank-correlation test.Results The GLS of all patients were higher at six months after delayed PCI than at one day prior to delayed PCI [(-15.39±7.80)％ vs (-12.44±8.38)％,t=14.398,P ＜ 0.001].The LPSS of myocardial perfusion in good,reduced and absent groups at one day prior to delayed PCI were (-2.64±5.60)％,(-6.19±6.87)％ and (-12.07±5.86)％,respectively.The LPSS of myocardial perfusion in good,reduced and absent groups at six months after delayed PCI were (-2.97 ± 4.93)％,(-11.38± 7.26)％ and (-15.82 ± 5.97)％,respectively.The myocardial LPSS of left ventricular segment with good or reduced perfusion was significantly higher at six months after delayed PCI (t=13.013,10.821,both P ＜ 0.001),but the LPSS of left ventricular segment with absent perfusion was similar to that of pre-PCI.Whether at one day prior to delayed PCI or six months after delayed PCI,there were significant differences in LPSS parameters among the three groups (at one day prior to delayed PCI,myocardial perfusion absent vs reduced or good,t=4.201 and 11.771,both P ＜ 0.001;myocardial perfusion reduced vs good,t=12.561,P ＜ 0.001;at six months after delayed PCI,myocardial perfusion absent vs reduced or good,t=9.714 and 15.646,both P ＜ 0.001;myocardial perfusion reduced vs good,t=9.254,P ＜ 0.001).The LPSS both at one day prior to delayed PCI and six months after delayed PCI in myocardial perfusion good group ＞ those of myocardial perfusion reduced group ＞ those of myocardial perfusion absent group.PSI was positively correlated with GLS at both one day prior to delayed PCI and six months after delayed PCI (r=0.69,0.72,both P ＜ 0.001).MPS was positively correlated with LPSS at both one day prior to delayed PCI and six months after delayed PCI (r=0.49 and 0.45,both P ＜ 0.001).Conclusion Myocardial perfusion before delayed PCI,monitored by MCE,is correlated well with myocardial systolic function,and may be used to predict the recovery of myocardial systolic function after delayed PCI.

Objective To discuss the training effect driven by clinical demand in a cross-sectional research, and provide evidence for specialized nursing training.Methods One month before the survey, 46 nurses received various forms of comprehensive training, including multimedia lectures, workshop, case analysis, group discussion, clinical demonstration teaching of ostomy wound, research operations training, risk assessment training of pressure ulcer and incontinence associated dermatitis, etc. Then theory examinations, picture analysis examinations and operation examinations were conducted before and after training based on the unified papers and operating criteria. Results After training, the score of theoretical examination, operating examination and picture analysis examination of 46 nurses were higher than that before the training (P<0.05). 89.1% of nurses thought that the training content was very good;93.5% of nurses were satisfied with the training form, and 95. 7% of nurses thought that this training was significant for clinical work. Conclusions The comprehensive training method based on clinical demand can effectively improve the professional theory and skills of nurses, thus it can be used as a effective training and assessment methods before the survey of pressure ulcer and incontinence dermatitis in comprehensive hospitals.