Clinical use of molecular information in the management of multiple endocrine neoplasia type 2A

Abstract. One hundred and ninety-seven members of 28 kindreds with multiple endocrine neoplasia type 2A (MEN 2A) were screened for RET proto-oncogene exon 10 and 11 mutations. Seventy-one known affected individuals had mutations of codons 609, 618, 620 or 634, whereas 53 unaffected individuals had no abnormalities. Nineteen out of 54 individuals of unknown status, mostly children, had RET mutations. Four of these children had thyroidectomy based on this analysis and were found to have C-cell abnormalities. We identified one false negative mutation analysis because of a codon 691 polymorphism. We conclude that RET mutational analysis is a cost-effective and accurate method for determination of gene carrier status in MEN 2A.     

膜联蛋白A2过表达在肝细胞癌发生发展中的作用

膜联蛋白A2与S100A10组装成异四聚体转位细胞膜从而激活相关信号通路或者Ser^25磷酸化以后进入细胞核参与DNA合成和细胞增殖,可调节血管形成、与肝细胞癌（HCC）形成与预后相关。本文综述了膜联蛋白A2的分子构成、功能及对HCC诊断和预后价值的研究进展。     

一个多发性内分泌腺瘤病2A型家系的RET原癌基因突变检测

目的 检测一个多发性内分泌腺瘤病(MEN)2A型家系中RET原癌基因的突变情况.方法 观察一个MEN2A家系成员的表型,并对与MEN相关的且点突变率较高的RET原癌基因第10和11外显子进行PCR产物直接DNA测序以了解其杂合性.结果 家系中4名家族成员均存在RET原癌基因第11外显子Cys(TGC)634Arg(CGC)错义突变和Gly(GGT)691Ser(AGT)的单核苷酸多态性,另有1名成员仅存在RET原癌基因Gly(GGT)691Set(AGT)的单核苷酸多态性.经B超检查发现其中2名成员双侧甲状腺及一侧肾上腺和一侧甲状旁腺有实性占位病变,1名成员双侧甲状腺及一侧肾上腺有实性占位病变,1名成员双侧甲状腺、双侧肾上腺和一侧甲状旁腺有实性占位病变,1名成员仅有甲状腺多发性小结节.另外有3名成员B超检查有异常,但无基因突变.结论 对MEN2A家系的基因分析证实RET原癌基因第11外显子634位密码子存在突变和(或)691位密码子存在单核苷酸多态性,对MEN2A能在基因水平作出诊断.     

Expression of PTEN, PPM1A and P-Smad2 in hepatocellular carcinomas and adjacent liver tissues

AIM: To investigate the expressions of PTEN, PPM1A and P-Smad2 in hepatocellular carcinoma (HCC) and their significance. METHODS: The expressions of PTEN, PPM1A and P-Smad2 in 31 HCC tissues, 25 adjacent liver tissues and 13 non-tumor liver tissues were detected by using Envision immunohistochemical technique. RESULTS: The positive expression (64.52%) and staining intensity (4.19 ± 3.31) of PTEN in the cytoplasm of HCC were significantly lower and weaker than those in the adjacent or non-tumor liver tissues (97.37%, 7.88 ± 0.93; 100%, 7.77 ± 0.93, respectively) (P < 0.05), and its staining intensity in the cytoplasm of HCC, which belongs to Edmondson pathologic grades Ⅱ-Ⅲ and above, was also lower than that of gradeⅠandⅠ-Ⅱ. Furthermore, its location in the nucleus or cytoplasm of liver cells was negatively correlated with the progression of liver disease (r = -0.339, P = 0.002); most of PPM1A might be only expressed in the nucleus of adjacent liver tissues, non-HCC tissues or Edmondson gradeⅠandⅠ-Ⅱ HCC, but it was mainly expressed in the cytoplasm of HCC with Edmondson grade ≥Ⅱ, weakly or negatively expressed in the nucleus (P < 0.05), and its location was negatively correlated with the progression of liver disease (r = -0.45, P = 0.0000). P-Smad2, which was mostly located in the nucleus and cytoplasm of gradeⅠ andⅠ-Ⅱ HCC, surrounding or non-tumor liver tissues, was only in the nucleus of HCC with Edmondson grade Ⅱ and above (P < 0.001), and its location was positively correlated with the disease progression (r = 0.224, P = 0.016). Spearman correlation analysis revealed that P-Smad2 was significantly negatively correlated with PTEN and PPM1A (r = -0.748, P = 0.000; r = -0.366, P = 0.001, respectively); and PTEN and PPM1A were positively correlated with HCC carcinogenesis (r = 0.428, P = 0.000). CONCLUSION: The aberrant location of expression and staining intensity of PTEN, PPM1A and P-Smad2 in HCC and their relationship might have an impact on the pathogenesis of HCC.     

HbS/βdel-thalassemia associated with high levels of hemoglobins A2 and F in a Turkish family

β-thalassemia and sickle cell disease (SCD) are common disorders in Turkey. Compound heterozygosity for these two disorders (β^S/β-thalassemia) is encountered frequently. In this report we present hematological and molecular data of two Turkish siblings with β^S/β^del-thalassemia caused by a 290 base pair (bp) deletion and associated with increased levels of hemoglobin A₂ (HbA₂) and hemoglobin F (HbF). Clinical analysis of the two patients showed a mild course of the disease. Haplotypic factors involved in increasing the levels of HbF were analyzed. The two patients showed no changes from the normal sequences at the XmnI site of Gγ-globin promoter and the (AT)_xT_y microsatellite 5′ to the β-globin mRNA cap site. The removal of the region between positions −125 to +78 relative to the β-globin gene mRNA cap site by the 290 bp deletion is thought to allow the β-locus control region to interact with the promoters of the δ- and γ-globin genes, leading to increased HbA₂ and HbF levels. Am. J. Hematol. 59:83–86, 1998. © 1998 Wiley-Liss, Inc.     

Assay discrepancy in mild haemophilia A due to a factor VIII missense mutation (Asn694Ile) in a large Danish family

Factor VIII gene analysis in a large consanguinous Danish family comprising 24 affected males and four homozygously affected females revealed an Asn694Ile mutation within the A2 domain. The factor VIII gene mutation led to a mild haemophilia A phenotype with factor VIII function displaying discordance between one-stage clotting and chromogenic two-stage assays. In one-stage assays, values ranged from 0.05 to 0.30 IU/ml (males) and from 0.19 to 0.29 IU/ml (homozygous affected females), whereas the chromogenic two-stage assay produced values of around only 50% of the one-stage result [0. 02-0.12 IU/ml (males); 0.06-0.10 IU/ml (females)]. The differences are suggested to be caused by the effect of the mutation on the active cleaved form of the factor (F)VIII protein. As the original amino acid (Asn) is conserved in all known FVIII A2 sequences, but not in ceruloplasmin, we suggest that Asn694 is involved in an A2-specific functional role. Examination of a homology model of the A domains predicts that the Asn694Ile mutation (i) results in the loss of two potential hydrogen-bonding interactions and (ii) hampers the integration of the bulky side-chain of Ile into the A2 domain core, probably causing an altered stability and/or folding of the protein. Interestingly, the disease in this Danish family was originally proposed to be von Willebrand-Jürgens disease. However, the current study rules out the co-existence of either von Willebrand's disease or the presence of the Normandy variant of von Willebrand factor (type 2N).     

Expression and clinical significance of S100A2 and p63 in esophageal carcinoma

AIM： TO investigate the expression and clinical significance of S100A2 mRNA and protein, p63 protein in esophageal squamous cell carcinoma （ESCC） and their roles in carcinogenesis and progression of esophageal carcinoma （EC）. METHODS： Immunohistochemical staining （S-P method） for S100A2 and p63 protein were performed in 40 samples of ESCC and 40 samples of normal esophageal mucosa. In situ hybridization （ISH） was used to detect the expression of S100A2 mRNA. RESULTS： Expression of S100A2 mRNA in ESCC was positive in 77.5% of samples, which was lower than that in normal mucosa （100%） by ISH （P = 0.002）. The expression level of S100A2 mRNA was closely related to differentiation and and node-metastasis （P = 0.012, P = 0.008）. Expression of $100A2 protein was positive in 72.5% of ESCC samples and expression of p63 protein was positive in 37.5% of ESCC samples, and was lower than that in normal mucosa （100%） （P = 0.000）. The expression of S100A2 protein was correlated with the differentiation and node-metastasis （P = 0.007, P = 0.001）, but no relationship was observed between the expression of p63 protein and clinical pathological manifestations. S100A2 protein was positively correlated with the expression of S100A2 mRNA, and negatively associated with the expression of p63 protein （P = 0.000, P = 0.002）. CONCLUSION： S100A2 and p63 protein both play important roles in the carcinogenesis of ESCC. An investigation into the combined expression of S100A2 and p63 may be helpful in early diagnosis and in evaluating the prognosis of ESCC.     

Coxsackievirus A2 Leads to Heart Injury in a Neonatal Mouse Model

Coxsackievirus A2 (CVA2) has emerged as an active pathogen that has been implicated in hand, foot, and mouth disease (HFMD) and herpangina outbreaks worldwide. It has been reported that severe cases with CVA2 infection develop into heart injury, which may be one of the causes of death. However, the mechanisms of CVA2-induced heart injury have not been well understood. In this study, we used a neonatal mouse model of CVA2 to investigate the possible mechanisms of heart injury. We detected CVA2 replication and apoptosis in heart tissues from infected mice. The activity of total aspartate transaminase (AST) and lactate dehydrogenase (LDH) was notably increased in heart tissues from infected mice. CVA2 infection also led to the disruption of cell-matrix interactions in heart tissues, including the increases of matrix metalloproteinase (MMP)3, MMP8, MMP9, connective tissue growth factor (CTGF) and tissue inhibitors of metalloproteinases (TIMP)4. Infiltrating leukocytes (CD45⁺ and CD11b⁺ cells) were observed in heart tissues of infected mice. Correspondingly, the expression levels of inflammatory cytokines in tissue lysates of hearts, including tumor necrosis factor alpha (TNF-α), interleukin-1beta (IL-1β), IL6 and monocyte chemoattractant protein-1 (MCP-1) were significantly elevated in CVA2 infected mice. Inflammatory signal pathways in heart tissues, including phosphatidylinositol 3-kinase (PI3K)-AKT, mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-κB), were also activated after infection. In summary, CVA2 infection leads to heart injury in a neonatal mouse model, which might be related to viral replication, increased expression levels of MMP-related enzymes and excessive inflammatory responses.     

Computer-supported indirect-form lifestyle-modification support program using Lifestyle Intervention Support Software for Diabetes Prevention (LISS-DP) for people with a family history of type 2 diabetes in a medical checkup setting: A randomized controlled trial

AimsTo investigate the effect of a computer-supported indirect-form lifestyle-modification program using Lifestyle Intervention Support Software for Diabetes Prevention (LISS-DP), as a clinically feasible strategy for primary prevention, on diet and physical activity habits in adults with a family history of type 2 diabetes.MethodsThis was a two-arm, randomized controlled trial: (1) lifestyle intervention (LI) group (n = 70); (2) control (n = 71). Healthy adults aged 30–60 years with a history of type 2 diabetes among their first-degree relatives were recruited. LI group received three times of lifestyle intervention using LISS-DP during six-month intervention period via mail.ResultsLifestyle intervention group showed significantly greater decrease in energy intake six months after baseline, compared to control (−118.31 and −24.79 kcal/day, respectively, p = 0.0099, Cohen's d = 0.22), though the difference disappeared 1 year after from baseline. No difference was found in physical activity energy expenditure.ConclusionsA computer-based, non-face-to-face lifestyle intervention was effective on dietary habits, only during the intervention period. Further examination of the long-term effects of such intervention and physical activity is required.     

The imbalance between NKG2A and NKG2D expression is involved in NK cell immunosuppression and tumor progression of patients with hepatitis B virus-related hepatocellular carcinoma

Background

Immunosuppression in a tumor microenvironment is associated with enhanced tumor progression. Natural killer group 2 (NKG2) family proteins, including inhibitory receptors and activators, can be used as attractive targets for immunotherapy of immune checkpoint inhibition. We further explore the expression level prognostic value of NKG2A and NKG2D in hepatitis B virus-related hepatocellular carcinoma (HBV-HCC).

Methods

This study was a prospective study involving 92 patients with HBV-HCC, 16 patients with HBV-related liver cirrhosis, 18 patients with CHB, and 38 healthy donors. We analyzed the expression and related functions of NKG2A, NKG2D, and the NKG2A/NKG2D ratio in the peripheral blood of patients with HBV-HCC and analyzed tumor progression. The tissue samples from patients with HBV-HCC were further used for multiple immunofluorescence and immunohistochemistry.

Results

In patients with HBV-HCC with tumor progression, the ratio of NKG2A/NKG2D is higher in NK cells and T cells. The Kaplan–Meier survival curve showed that the NKG2A/NKG2D ratio on NK cells could predict tumor progression in patients with HBV-HCC, and that an increase in this ratio was associated with inhibition of NK cell function. The Cancer Genome Atlas (TCGA) database was further used to verify that the higher the NKG2A/NKG2D ratio, the shorter the progression-free survival of patients with HCC, and the more likely the immune function was suppressed.

Conclusions

The imbalance between NKG2A and NKG2D of NK cells is involved in NK cell immunosuppression, and the increase of the NKG2A/NKG2D ratio is related to the tumor progression of HBV-HCC.     

Adiponectin receptors gene expression and insulin sensitivity in non-diabetic Mexican Americans with or without a family history of Type 2 diabetes

Aims/hypothesis The recent discovery of two adiponectin receptors (AdipoR1 and AdipoR2) will improve our understanding of the molecular mechanisms underlying the insulin-sensitising effect of adiponectin. The aim of this study was to determine for the first time whether skeletal muscle AdipoR1 and/or AdipoR2 gene expression levels are associated with insulin resistance.Methods Using RT-PCR and northern analysis we measured AdipoR1 and AdipoR2 gene expression in skeletal muscle from healthy Mexican Americans with normal glucose tolerance who had (n=8) or did not have (n=10) a family history of Type 2 diabetes.Results Gene expression profiling indicated that the AdipoR1 and AdipoR2 isoforms are highly expressed in human skeletal muscle, unlike in mice where AdipoR2 expression was highest in the liver, and AdipoR1 was highest in skeletal muscle. In the study subjects, the expression levels of AdipoR1 (p=0.004) and AdipoR2 (p=0.04), as well as plasma adiponectin concentration (p=0.03) were lower in people with a family history of Type 2 diabetes than in those with no family history of the disease. Importantly, the expression levels of both receptors correlated positively with insulin sensitivity (r=0.64, p=0.004 and r=0.47, p=0.048 respectively).Conclusions/interpretation Collectively, these data indicate that both isoforms of the adiponectin receptor play a role in the insulin-sensitising effect of adiponectin.Abbreviations AdipoR1 adiponectin receptor-1 - AdipoR2 adiponectin receptor-2 - AMKP adenosine 5-monophosphate-activated protein kinase - FH+ with a family history of Type 2 diabetes - FH– with no family history of Type 2 diabetes - PPAR- peroxisome proliferator-activated receptor-     

Adverse reactions to BNT162b2 mRNA COVID-19 vaccine in medical staff with a history of allergy

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) vaccination is progressing globally. Several adverse reactions have been reported with vaccination against COVID-19. It is unknown whether adverse reactions to COVID-19 vaccination are severe in individuals with allergies.MethodsWe administered the COVID-19 vaccine to the medical staff at Yamagata University Hospital from March to August 2021. Subsequently, we conducted an online questionnaire-based survey to investigate the presence of allergy and adverse reactions after vaccination and examine the association between allergy and adverse reactions after immunization.ResultsResponses were collected from 1586 to 1306 participants after the first and second administration of the BNT162b2 mRNA COVID-19 vaccine, respectively. Adverse reactions included injection site pain, injection site swelling, fever, fatigue or malaise, headache, chills, nausea, muscle pain outside the injection site, and arthralgia. The frequency of some adverse reactions and their severity were higher, and the duration of symptoms was longer in participants with allergies than in those without allergies. Although several participants visited the emergency room for treatment after the first and second vaccinations, no participant was diagnosed with anaphylaxis.ConclusionsThis study suggests that the frequency and severity of adverse reactions after injection of BNT162b2 mRNA COVID-19 vaccine were higher in individuals with allergy; however, no severe adverse reactions such as anaphylaxis or death were observed. These results indicate that individuals with allergic histories may tolerate the BNT162b2 mRNA COVID-19 vaccine.     

食管鳞癌组织中S100A4和MMP2蛋白表达与临床病理学特征的关系

目的:探讨食管鳞癌组织和癌旁正常黏膜组织中S100A4和MMP2的表达及其与食管鳞癌临床病理因素之间的关系.方法:应用SP免疫组织化学技术检测100例食管鳞癌及其癌旁正常黏膜组织中S100A4和MMP2蛋白的表达.结果:食管鳞癌组织中S100A4和MMP2蛋白阳性率分别为52.00%和67.00%,明显高于癌旁正常黏膜组织26.00%和3 1.00%(P<0.01).二者在食管鳞癌中的表达呈正相关,且与癌组织的分化程度、浸润深度、淋巴结转移密切相关(P<0.05).结论:S100A4可能通过对MMP-2蛋白的调控在食管鳞癌的发生、发展及食管鳞癌的早期侵袭转移中起关键作用,有望成为评估食管癌预后的一个新的标志物.