Current understanding and progress of research on isovaleric acidemia / 中华医学遗传学杂志

Isovaleric acidemia is a type of organic acidemia for which the earliest definite diagnosis was attained. It features an autosomal recessive inheritance, with the onset of age varying from newborn to adulthood. The clinical manifestations are complex and variable, which include feeding difficulty, vomiting, lethargy, coma, metabolic acidosis, sweaty feet odor and mental retardation. The mortality and mobility rates of isovaleric acidemia are quite high, and early diagnosis and rational treatment can significantly improve the prognosis. This article has provided a summary for the current understanding and research progress on isovaleric acidemia.

Correlation between serum insulin-like growth factor 1, thyroid stimulating hormone, degree of insulin resistance and thyroid nodule imaging reporting and data system in patients with type 2 diabetes mellitus / 中国医师进修杂志

Objective:To investigate the correlation between serum insulin-like growth factor 1 (IGF-1), thyroid stimulating hormone (TSH), degree of insulin resistance and thyroid nodule imaging reporting and data system (TI-RADS) grading in patients with type 2 diabetes mellitus (T2DM).Methods:The clinical data of 120 patients with T2DM from February 2020 to November 2021 in Kunshan Hospital of Integrated Traditional Chinese and Western Medicine were retrospectively analyzed. Among them, 56 patients had no thyroid nodules (non-thyroid nodule group), all patients were TI-RADS grade 1; 64 patients had thyroid nodules (thyroid nodule group), including 7 cases of TI-RADS grade 2, 12 cases of TI-RADS grade 3, 20 cases of TI-RADS grade 4, and 25 cases of TI-RADS grade 5. The levels of IGF-1 and TSH were measured by automated biochemical analyzer, the homeostatic model assessment insulin resistance index (HOMA-IR) was calculated. Spearman method was used for correlation analysis; multivariate Logistic regression was used to analyze the independent risk factors of TI-RADS grading in patients with T2DM combined with thyroid nodules; the receiver operating characteristic (ROC) curve was drawn to analyze the efficacy of IGF-1, TSH and HOMA-IR in predicting TI-RADS grading in patients with T2DM combined with thyroid nodules.Results:The IGF-1, TSH and HOMA-IR in thyroid nodule group were significantly higher than those in non-thyroid nodule group: (185.35 ± 45.08) ng/L vs. (168.36 ± 30.25) ng/L, (2.98 ± 0.85) mU/L vs. (2.69 ± 0.35) mU/L and 3.25 ± 0.75 vs. 2.95 ± 0.44, and there were statistical differences ( P<0.05 or <0.01). In patients with T2DM combined with thyroid nodules, with the increase of TI-RADS classification, the IGF-1, TSH and HOMA-IR gradually increased, and there were statistical differences ( P<0.05). Spearman correlation analysis result showed that the levels of IGF-1, TSH and HOMA-IR were positive correlation with TI-RADS grading ( r = 0.918, 0.906 and 0.920; P<0.05). Multivariate Logistic regression analysis result showed that the IGF-1, TSH and HOMA-IR were independent risk factors for TI-RADS grading in patients with T2DM combined with thyroid nodule ( OR = 1.684, 1.044 and 1.851; 95% CI 0.674 to 6.665, 0.032 to 0.055 and 1.212 to 2.298; P<0.01 or <0.05). ROC curve analysis result show that the area under the curve of IGF-1, TSH and HOMA-IR for predicting the TI-RADS grading patients with T2DM combined with thyroid nodule were 0.946, 0.983 and 0.975, with all sensitivity of 100.00%, and specificity of 82.14%, 91.07% and 89.29%. Conclusions:There is a correlation between IGF-1, TSH, HOMA-IR and TI-RADS grading in patients with T2DM combined with thyroid nodule, which has some guiding value for clinical monitoring of thyroid nodule changes.

Interpretation on the international consensus of the pathological diagnosis of unilateral primary aldosteronism / 中华内分泌代谢杂志

Unilateral primary aldosteronism (UPA) is a common form of PA that is surgically curable by adrenalectomy of the overactive gland. Pathological evaluation of resected adrenals is crucial in the diagnosis of UPA, and its subsequent treatment and follow-up as well. Histomorphological evaluation is the basis for the pathological diagnosis of UPA, and the wide use of aldosterone synthase immunohistochemical staining in recent years has greatly improved the pathological diagnosis of UPA. However, there is a lack of standardized nomenclature and diagnostic criteria. Therefore, consensus on the histopathologic diagnosis of UPA were developed by an international group of pathologists led by Tracy Ann Williams, and published in J Clin Endocrinol Metab, 2021. This article will elaborates on the key points in the consensus to advance the understanding and overall improve clinical mangement of UPA.

Effects of delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots / 浙江大学学报·医学版

OBJECTIVE@#To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide evidence for improving accurate and reliable detection by tandem mass spectrometry.@*METHODS@#A total of 1 254 616 newborn DBS samples in Newborn Screening Center of Zhejiang Province were delivered and stored at room temperature (group A, @*RESULTS@#The concentrations of amino acids and carnitines in the three groups were skewed, and the differences in amino acid and carnitine concentrations among groups were statistically significant (all @*CONCLUSIONS@#Cold-chain logistics system and storage in low temperature and low humidity can effectively reduce degradation of some amino acids and carnitines in DBS, improve the accuracy and reliability of detection, and thus ensures the quality of screening for neonatal metabolic diseases.

NKT cells in liver diseases / 医学前沿

Natural killer T cells are innate-like and tissue-resident lymphocytes, which recognize lipid antigens and are enriched in the liver. Natural killer T cells play important roles in infections, tumors, autoimmune diseases, and metabolic diseases. In this study, we summarize recent findings on biology of natural killer T cells and their roles in hepatitis B virus and hepatitis C virus infection, autoimmune liver diseases, alcoholic liver disease, nonalcoholic fatty liver disease, and hepatocellular carcinoma. Controversial results from previous studies are discussed, and indicate the dynamic alteration in the role of natural killer T cells during the progression of liver diseases, which might be caused by changes in natural killer T subsets, factors skewing cytokine responses, and intercellular crosstalk between natural killer T cells and CD1d-expressing cells or bystander cells.

Analysis of Inflammatory cytokine expression in cluster headache / 中国神经精神疾病杂志

Objective To analysis the expression levels of inflammatory cytokines in patients with cluster headache during headache attack period and intermittent period using antibody chips to explore the role of inflammatory cytokines in the pathogenesis of cluster headache. Methods Blood samples from 6 patients with cluster headache were collected during headache attack period and intermittent period. Samples were then centrifugated and stored at - 80 degrees refrigerator. Samples were further labeled with biotin and reacted with antibody chips against 40 major inflammatory cytokines. The target proteins were conjugated with streptomycin antibody labeled with infrared fluorescent agent, and signals were transformed to images by Licor-odyssey scanner. Results In pairwise comparisons, the levels of some inflammatory cytokines were significantly increased during attacks compared to intermittent period including interleukin-1β(44.18 vs. 68.46, P<0.05), interleukin-6(23.08 vs. 36.40, P<0.05), interleukin-8(151.87 vs. 328.12, P<0.05), interleukin-13(23.93 vs. 38.87, P<0.05), monoeyte chemoattraetant protein (454.80 vs. 725.75, P<0.05) and macrophage inflammatory protein-1β (265.08 vs. 515.74, P<0.05). Conclusion Inflammatory cytokines may play an important role in the pathogenesis of cluster headache. However, the mechanism needs further investigation.

miR-30c regulates proliferation, apoptosis and differentiation via the Shh signaling pathway in P19 cells

MicroRNAs (miRNAs) are small, non-coding single-stranded RNAs that suppress protein expression by binding to the 3′ untranslated regions of their target genes. Many studies have shown that miRNAs have important roles in congenital heart diseases (CHDs) by regulating gene expression and signaling pathways. We previously found that miR-30c was highly expressed in the heart tissues of aborted embryos with ventricular septal defects. Therefore, this study aimed to explore the effects of miR-30c in CHDs. miR-30c was overexpressed or knocked down in P19 cells, a myocardial cell model that is widely used to study cardiogenesis. We found that miR-30c overexpression not only increased cell proliferation by promoting cell entry into S phase but also suppressed cell apoptosis. In addition, we found that miR-30c inhibited dimethyl sulfoxide-induced differentiation of P19 cells. miR-30c knockdown, in contrast, inhibited cell proliferation and increased apoptosis and differentiation. The Sonic hedgehog (Shh) signaling pathway is essential for normal embryonic development. Western blotting and luciferase assays revealed that Gli2, a transcriptional factor that has essential roles in the Shh signaling pathway, was a potential target gene of miR-30c. Ptch1, another important player in the Shh signaling pathway and a transcriptional target of Gli2, was downregulated by miR-30c overexpression and upregulated by miR-30c knockdown. Collectively, our study revealed that miR-30c suppressed P19 cell differentiation by inhibiting the Shh signaling pathway and altered the balance between cell proliferation and apoptosis, which may result in embryonic cardiac malfunctions.

Effects of microRNA -30c knockdown on proliferation and differentiation of P19 cells / 中华实用儿科临床杂志

Objective To explore the effects of microRNA(miRNA)- 30c knockdown on proliferation,diffe-rentiation of P19 cells. Methods miRNA - 30c knockdown plasmid(miRNA - 30c knockdown group)or no - load vector(negative control group)was transfected into P19 cells by lipo2000 and stable cell lines were selected by Blastici-din;Dual luciferase reporter gene system was used to confirm miRNA - 30c knockdown. Cell counting kit - 8(CCK - 8) assay was adopted to detect cell proliferation activity. An inverted microscope was used to observe morphological chan-ges of P19 cell differentiation. Cells were induced to differentiated to myocardiocyte with dimethyl sulfoxide(DMSO). Differentiation marker genes including cTnT,NKX2. 5,GATA4 relative mRNA expression levels were detected with real - time quantitative polymerase chain reaction,respectively. Results Observation of green fluorescent protein ex-pression under a fluorescence microscope indicated similar transfection efficiencies,and miRNA - 30c knockdown re-leased the activity of target gene Gli2. As a result,miRNA - 30c knockdown vector was constructed successfully(P ﹤0. 001). During differentiation of mouse P19 cells into myocardial cells,the beating cell clusters in miRNA - 30c knockdown cells were much lower than those in the control cells,and cTnT,NKX2. 5,GATA4 in miRNA - 30c knock-down cells showed significantly lower expression than those in the control cells( all P ﹤ 0. 05). Conclusions miRNA - 30c inhibits the P19 cell proliferation and differentiation. This study gives us a new insight of heart develop-ment and we need more efforts on exploring the deep function of heart diseases.

Effect of shRNA silenced aromatic hydrocarbon receptor on WNT signaling pathway during the differentiation of cardiac myocytes / 临床儿科杂志

Objective To explore the effect of shRNA silenced aromatic hydrocarbon receptor (AHR) on WNT signaling path-way during the differentiation of P19 cells into cardiac myocytes. Methods The eukaryotic expression vector of mouse AHR gene was designed and constructed. The interference plasmid was transfected into P19 cell and the positive stains to AHR gene silencing were screened by G418. The mRNA expression of important genes GSK3βandβ-catenin were evaluated by real-time fluorescent quantita-tive PCR during the differentiation of P19 cells. Results The constructed AHR-shRNA plasmid significantly inhibited the expression of AHR gene. Along with the differentiation of P19 cell into cardiac myocytes, in the interference group the expression ofβ-catenin gene was lower whereas the expression of GSK3βgene was elevated than those of control group with significant differences (all P<0.01). Conclusions The interference of AHR gene expression can regulate WNT signaling pathway in the development of heart.