Dias-Logan syndrome, also known as intellectual developmental disorder with persistence of fetal hemoglobin (HbF), or BCL11A-related intellectual developmental disorder, is an extremely rare neurogenetic disorder characterized by intellectual disability (ID), delayed psychomotor development, variable dysmorphic features, and asymptomatic persistence of fetal hemoglobin. The prevalence and incidence of this condition are currently unknown. We report an 8-year-old Han Chinese male patient with Dias-Logan syndrome who carries a de novo heterozygous pathogenic variant, c.1078dupC (p.Leu360Profs*212), in the BCL11A gene, leading to ID and γ-globin suppression, identified through trio-based whole exome sequencing (trio-WES). All his blood parameters were normal except for an elevated HbF level, which was 19.9% of total hemoglobin. Given the negative family history for ID, epilepsy, and alcohol consumption, de novo inheritance was presumed. Consequently, trio-WES analysis (parents and child) was conducted as it can identify potential new causal variants in the offspring. So far, a comprehensive understanding of the phenotypic spectrum of Dias-Logan syndrome and the impact of genotypic variation on disease severity is still lacking. Therefore, our case report enriches the existing literature on the clinical spectrum and genotype–phenotype correlations of BCL11A-related syndrome and provides some helpful information for diagnosis, management, and genetic counseling.

Dias-Logan syndrome, also known as intellectual developmental disorder with persistence of fetal hemoglobin (HbF), or BCL11A -related intellectual developmental disorder, is an extremely rare neurogenetic disorder characterized by intellectual disability (ID), delayed psychomotor development, variable dysmorphic features, and asymptomatic persistence of fetal hemoglobin. The prevalence and incidence of this condition are currently unknown. We report an 8-year-old Han Chinese male patient with Dias-Logan syndrome who carries a de novo heterozygous pathogenic variant, c.1078dupC (p.Leu360Profs*212), in the BCL11A gene, leading to ID and γ-globin suppression, identified through trio-based whole exome sequencing (trio-WES). All his blood parameters were normal except for an elevated HbF level, which was 19.9% of total hemoglobin. Given the negative family history for ID, epilepsy, and alcohol consumption, de novo inheritance was presumed. Consequently, trio-WES analysis (parents and child) was conducted as it can identify potential new causal variants in the offspring. So far, a comprehensive understanding of the phenotypic spectrum of Dias-Logan syndrome and the impact of genotypic variation on disease severity is still lacking. Therefore, our case report enriches the existing literature on the clinical spectrum and genotype–phenotype correlations of BCL11A -related syndrome and provides some helpful information for diagnosis, management, and genetic counseling.

Objective: Polycystic ovarian syndrome (PCOS) is a prevalent gynecologic disorder, often associated with abnormal follicular development. Cangfu Daotan decoction (CFD) is a traditional Chinese medicine formula that is effective in alleviating PCOS clinically, but the specific mechanism remains unclear. Forkhead box K1 (FOXK1) is associated with cellular function. This study aimed to explore the effects of CFD and FOXK1 on PCOS.Methods: High-fat diet and letrozole were combined to establish PCOS rat models. Next, primary GCs were extracted from those PCOS rats. Then, GC cells were transfected with si-FOXK1 or oe-FOXK1. CFD-contain serum was prepared, and experiments were conducted to investigate the regulation of FOXK1 by CFD.Results: FOXK1 was highly expressed in GCs of PCOS rats. Further investigation revealed that FOXK1 overexpression resulted in inhibition of proliferation and DNA synthesis, along with promotion of apoptosis and autophagy in GCs. Additionally, it was found that FOXK1 promoted the expressions of the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway-related proteins. Interestingly, treatment with CFD reversed all the effects of FOXK1 overexpression in GCs. Conclusion: This study demonstrated that CFD exerted a protective role in PCOS by inhibiting FOXK1, which provided a research basis for the application of CFD in PCOS, and suggested that FOXK1 is a novel therapeutic target in PCOS treatment.

Objective Polycystic ovarian syndrome (PCOS) is a common disorder that leads to infertility in reproductive-aged females. HOTAIR is highly expressed in various gynecological diseases and is associated with a poor prognosis. We aimed to explore the role of HOTAIR in PCOS. Methods First, PCOS rats were induced using dehydroepiandrosterone and then treated with si-HOTAIR. Next, HOTAIR mRNA expression and serum hormone levels were detected. HE staining was applied to observe estrus cycle, ovarian morphology and count the number of follicles. Apoptosis in the ovary was detected by TUNEL. Thereafter, ovarian granulosa cells (GCs) were isolated from PCOS rats, transfected with si-HOTAIR and treated with LY294002 (Akt inhibitor) or IGF-1. CCK-8 and flow cytometry assays were used to evaluate cell viability and apoptosis. IGF-1, apoptosis- and PI3K/Akt pathway-associated protein expressions in ovary and GCs were also detected. Results In in vivo experiments, si-HOTAIR decreased serum T, E-2 and LH levels but increased FSH level, restored estrus cycle, ovarian morphology and inhibited apoptosis of ovary in PCOS rats. Meanwhile, in vitro assays showed that si-HOTAIR upregulated the viability but inhibited the apoptosis of PCOS GCs. Furthermore, both in vivo and in vitro assays revealed that si-HOTAIR increased Bcl-2 expression but suppressed Bax, Bad, IGF-1 expressions and PI3K, AKT phosphorylation. However, the aforementioned effects of si-HOTAIR in vitro were further enhanced by LY294002 and partially reversed by IGF-1. Conclusions HOTAIR knockdown improved PCOS, and the mechanism may relate to IGF-1-mediated PI3K/Akt pathway, indicating HOTAIR may be a novel therapeutic target for PCOS.

目的 采用数据挖掘的方法,通过中医辅助传承系统软件,研究中医药治疗反复胚胎移植失败的方药规律.方法 检索并筛选中国期刊全文数据库(CNKI)、维普、万方数据服务平台、中国生物医学文献、PubMed数据库近20年来有关中医药辨治反复胚胎移植失败的期刊论文,建立处方药数据库,并导入中医传承辅助平台V3.0,用频数分析、聚类分析、关联规则等方法进行数据挖掘,总结常用药物、组合、类方,分析药物及类方特点.结果 检索文献数据库之后,共筛选出文献103篇,包含处方112条.使用频次较多的中药依次为:补虚药,活血化瘀药,清热药;112首处方中用药四气以温、平为主,五味以甘味、苦味、辛味为主,归经以肝经、肾经、脾经为主;通过关联规则分析共挖掘出10条常用药物组合;经聚类分析,获得8条核心类方.结论 从中医药的观点来看,反复胚胎移植失败病位在肝、肾、脾,病性以虚为主,又见虚中夹实,治疗以补益肝肾、益气健脾、活血祛瘀为主.

目的 观察阴阳平衡针、枸橼酸氯米芬片、来曲唑,三种促排卵治疗方法下,对排卵障碍患者助孕结局的影响,为临床排卵障碍患者的治疗提供参考依据.方法 选取2019年12月至2020年12月于本科室就诊的共200例排卵障碍患者为研究对象,分别根据患者意向,进行分组治疗.随访时间从治疗开始,直至早孕12W结束.记录助孕过程当中,相关参数(单周期获得优势卵泡个数、单周期卵泡排出数、周期妊娠率、早期流产率).结果 三组分组患者进行数据比对,单周期获得优势卵泡个数、单周期卵泡排出数方面,三个组别未见明显差异.早期妊娠率,A组49.3％,明显优于其他两个组.早期流产率,A组7.5％,明显优于其他两个组,差异具有统计学意义.结论 阴阳平衡针对排卵障碍患者促进卵泡发育,提高妊娠率、降低流产率方面效果最好,来曲唑组在促进卵泡发育率也较高,但妊娠后早期流产率略高于阴阳平衡针组,枸橼酸氯米芬片组在卵泡排出、妊娠率及早期流产率方面均劣于其他两组.

Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder prevalent in females of reproductive age; insulin resistance (IR) is the major pathogenic driver. Pharmacology is a basic option for PCOS therapy; traditional Chinese medicine (TCM), as a significant part of complementary and alternative medicine, has a long history in the clinical management of PCOS. Cangfudaotan decoction (CFD) has been used clinically for gynaecological diseases especially PCOS. In this study, first, chemical components in CFD were clarified using UPLC-Q/TOF-MS analysis. Then, an animal model of PCOS was established, granular cells were also isolated from the rats with PCOS, and CFD was administrated at different dosages in PCOS rats and granular cells, to investigate the therapeutic effect and mechanisms of CFD for PCOS treatment. The result showed that CFD treatment is effective in PCOS rats and granulosa cells. CFD was able to improve IR, restore the serum hormone levels, inhibit the inflammatory cytokines in PCOS rat, and alleviate ovary morphological injury and apoptosis in PCOS rats. In granulosa cells of PCOS, the result showed that the cell viability was improved, and cell apoptosis was inhibited after CFD administration. Further experiments suggested that CDF improves IR, follicular development, cell apoptosis, and inflammatory microenvironment, and this was associated to the regulation of IGF-1-PI3K/Akt-Bax/Bcl-2 pathway-mediated gene expression. Given that CFD sufficiently suppresses insulin resistance and improves follicular development in this study, exploring these mechanisms might help to optimize the therapeutic treatment of CFD in PCOS patients.

Background Polycystic ovary syndrome (PCOS) is a common and complex female endocrine disorder. The pathogenesis of PCOS is not fully elucidated. Cangfu Daotan Decoction is a kind of traditional Chinese medicine prescription widely used for PCOS patients.Methods In this study, we treated two PCOS patients with Cangfu Daotan Decoction (CDD) and both patients presented favorable responses to the treatment. We performed RNA-seq to explore the underlying molecular mechanism.Results After data analysis, we found 4 differential expressed genes including S1PR1 , which has been found to be involved in islet β-cell proliferation and cell apoptosis in diabetic mouse models. We also identified 28 differential expressed transcripts including NFATC2 , CD14 and FCGR3A , which are involved in many immune processes.Conclusions After GO enrichment analysis, we found that pathways including viral gene expression and immune response are involved in the efficacy of CDD treatment. Our study provides new evidence to better understand the pathogenesis of PCOS.

目的 探讨苍附导痰汤加减对痰湿型多囊卵巢综合征(PCOS)患者的性激素水平及排卵率的影响.方法 选择痰湿型PCOS患者64例,随机分为2组,A组31例,均实施单纯苍附导痰汤加减汤药口服治疗;B组33例,给予单纯穴位针刺疗法.治疗前后分别测定患者性激素水平及监测自发排卵情况.结果 在连续治疗3个月后,A组中,治疗后LH/FSH比值及T均低于治疗前,尤其是LH/FSH比值明显低于治疗前(P＜0.05).B组中,治疗后LH/FSH比值及T略低于治疗前,但与治疗前的差异比较无统计学意义.2组治疗前后的排卵率相比较,治疗后,排卵率均有升高,A组治疗前后的排卵率相比较,差异有统计学意义(P＜0.05).结论 单纯苍附导痰汤加减口服比单纯针刺治疗痰湿型PCOS效果显著,能够有效改善性激素水平,提高排卵率.

目的 研究女性尿道支撑组织成纤维细胞中雌激素受体β (ERβ) 同分异构体的表达, 从而获得雌激素受体β同分异构体在女性压力性尿失禁中的作用及机制.方法 取阴道前壁组织作为研究标本, 并取正常对照组标本及患者实验组标本各5例.体外构建女性尿道支撑组织成纤维细胞模型.用免疫组织化学、RT-PCR、ELISA以及Western blot等方法检测尿道支撑组织中ERβ及ERβ同分异构体的表达水平.结果 免疫组化检测的结果发现, 与对照组相比, 实验组的ERβ表达存在一定程度的下调.Western blot的结果显示, 实验组中的ERβ蛋白含量明显低于正常组组织中的ERβ蛋白含量.用RT-PCR方法检测ERβ各个亚型表达之后发现, 与对照组相比, 实验组的ERβ1、ERβ2、ERβ3、ERβ5 mRNA均下调表达.结论 雌激素受体β同分异构体的表达下调与女性压力性尿失禁的发病相关联.

Objective To detect the level changes and its significance of serum VEGF,TGF-31 in recurrent spontaneous abortion.Methods 120 cases of recurrent spontaneous abortion (RSA) were selected as the research objects with 60 cases in primary RSA group,secondary RSA group and the control group respectively.The levels of VEGF and TGF-β1 in serum were measured.Results The levels of VEGF and TGF-β1 in the primary group were (2.41±0.52) ng/ml and (10.58 ±1.49)ng/ml,while that were (2.25 ± 0.63) ng/ml and (9.75 ± 1.70) ng/ml in the secondary RSA group,and (0.92 ± 0.39) ng/ml and (12.20 ± 1.88) ng/ml in the control group,there was statistical significance on the difference among the three groups (P ＜ 0.01);which were (2.23 ± 0.48) ng/ml,(11.06 ± 1.24) ng/ml at the early stage of the primary RSA group and (2.61 ± 0.50)ng/ml,(10.04 ± 1.57) ng/ml at the advanced stage in the primary RSA group;that were (2.03 ±0.69) ng/ml,(10.37 ± 1.50) ng/ml at the early stage and (2.48 ±0.49) ng/ml,(9.13 ± 1.70) ng/ml at the advanced stage in the secondary RSA group,there was statistical significance on the difference between the two groups(P ＜ 0.01);the serum concentrations of VEGF and TGF-β1 were positively correlated with RSA grading in the primary RSA group(r =0.64,r =0.65,P ＜ 0.01);and that in the secondary RSA group was also positively correlated with RSA grading(r =0.63,r =0.64,P ＜ 0.01).Conclusion The serum levels of VEGF,TGF-β1 can reflect the inflammation of recurrent spontaneous abortion.

目的 研究中医、理疗系统疗法对盆腔炎性疾病后遗症患者榆卵管通液压力变化的研究.方法 选取我科收治的盆腔炎性疾病后遗症患者150例,根据患者意愿,分为两组,每组75例患者.实验组子中医、理疗进行系统治疗,对照组仅中药口服,不加理疗等治疗手段.3周为一个疗程,治疗结束后,对比治疗前后患者子宫输卵管通液压力数值.结果 治疗前后,两组患者通液压力值均有所下降.实验组下降幅度较大,差异有统计学意义(P＜0.05),对照组无统计学意义(P＞0.05).结论 中医、理疗系统疗法能有效治疗盆腔炎性疾病后遗症患者输卵管粘连、通而不畅等问题,该法在临床上具有指导意义,值得临床推广.

In this study, we propose a finite element analysis of the complete cervical spine with straightened and normal physiological curvature by using a specially designed modelling system. An accurate finite element model is established to recommend plausible approaches to treatment of cervical spondylosis through the finite element analysis results. There are few reports of biomechanics influence of the straightened cervical curve. It is difficult to measure internal responses of cervical spine directly. However, the finite element method has been reported to have the capability to quantify both external and internal responses to mechanical loading, such as the strain and stress distribution of spinal components. We choose a subject with a straightened cervical spine from whom to collect the CT scan data, which formed the basis of the finite element analysis. By using a specially designed modelling system, a high quality finite element model of the complete cervical spine with straightened curvature was generated, which was then mapped to reconstruct a normal physiological curvature model by a volumetric mesh deformation method based on discrete differential properties. Then, the same boundary conditions were applied to do a comparison. The result demonstrated that the active movement range of straightened cervical spine decreased by 24-33 %, but the stress increased by 5-95 %. The stress was concentrated at the facet joint cartilage, uncovertebral joint and the disk. The results suggest that cervical lordosis may have a direct impact on cervical spondylosis treatment. These results may be useful for clinical treatment of cervical spondylosis with straightened curvature.

目的 颈椎后纵韧带骨化症(OPLL)是一种病因未明的骨科疾病,好发于日本和亚洲人群.近年来的研究认为,OPLL为一种遗传因素和环境因素共同作用所致的复杂疾病.我们选取了50例浙江地区汉族颈椎后纵韧带骨化症患者,以100例正常个体作为对照组,对已经有文献报道的与OPLL发病可能相关联的9个基因(COL11 A2、COL6A1、COL17A1、ENPP1、TGF-β3、BMP-2、BMP-4、LEPR、ESR1)的11个多态性SNP位点进行了基因分型,探讨相关性.方法 PCR+ Sanger DNA测序,将各个位点基因型的分布信息进行统计分析.结果 未发现所研究的OPLL患者群体与上述9个基因的11个多态性SNP位点存在显著的相关性.结论 OPLL易感基因的鉴定,需要大样本量的研究以及全外显子组测序、连锁分析等方法的相结合,方能得出最终的结论.

Objective To evaluate the clinical significance of plasma D-dimmer levels in patients with severe trauma. Methods Four hundred and eleven patients with severe trauma (trauma group) and 400 healthy subjects (control group) were in-cluded in this study. The plasma D-dimmer levels were measured by col oidal gold immune infiltration method in controls and in trauma patients at 24 and 96h after surgery respectively. Results The D-dimmer positive rate and plasma levels at 24 and 96h after surgery in trauma patients were significantly higher than those of normal controls (P<0.01). Traumatic deep vein thrombosis (TDVT) developed in 38 patients (9.2%), whose 24 and 96 h D-dimmer levels were significantly higher than that of control group (both P<0.01) and 373 trauma patients without TDVT developed (P<0.05 or 0.01). The rate of high D-dimmer levels at 24h in TDVT patients was significantly higher than that in non-TDVT trauma patients (65.8%, 1.9%, P<0.01). Conclusion High plasma D-dimmer level may indicate the development of traumatic deep vein thrombosis in patients with severe trauma.

Cornelia de Lange syndrome (CdLS; OMIM: 122470) is characterized by distinctive facial features, growth retardation, hirsutism, and upper limb reduction defects. Craniofacial features manifest as synophrys, arched eyebrows, long thick eyelashes, a small upturned nose, small widely-spaced teeth, and microcephaly. The intelligence quotient (IQ) is usually below the normal level. More phenotypes are frequently found, such as cardiac septal defects, gastrointestinal dysfunction, hearing loss, myopia, and cryptorchidism or hypoplastic genitalia. Some individuals suffering from milder forms of CdLS have less severe growth, cognitive, and limb involvement, but often have typical facial features.1 The prevalence of CdLS is estimated to be 1.6 to 2.2/100 000 live births, without racial variation.2 Most cases are sporadic, and very few familial cases have been reported. Approximately 50%-60% of CdLS cases are caused by mutations in the Nipped-B-like (NIPBL) gene, which encode two isoform products termed delangin-A and delangin-B. Mutations in the Structural Maintenance of Chromosomes 1A (SMC1A) and 3 (SMC3) genes account for ˜5% of CdLS cases and have been shown to cause a consistently mild phenotype.3 All three genes encode components of the sister chromatid cohesin complex. A diagnosis of CdLS is based on not only clinical findings, but also molecular genetic testing. Here, we report the first molecular analysis of a young patient diagnosed with CdLS in China. A 3-year-old girl with multiple congenital anomalies was referred to our clinic. The patient presented typical facial features: arch-like confluent eyebrows, long thick eyelashes, low anterior and posterior hairlines, anteverted nares, collapsed nasal bridge, downturned corners of the mouth, thin lips and low-set ears. She also had microcephaly, small hands with the right missing one digit and the left missing two, hirsutism, and a short neck (Figure 1A and 1B). The girl had developmental delay and mental retardation. She was the first child of her parents who denied a consanguineous relationship, following a first, uneventful pregnancy. There was no positive history of any deformity in the parents' families.Figure 1.: Characteristic features of the CdLS patient and sequencing of the NIPBL gene. A: Arch-like confluent eyebrows, long thick eyelashes, anteverted nares, collapsed nasal bridge, downturned corners of the mouth, thin lips and low-set ears. B: Upper-limb abnormalities. C: Chromographs of the partial cDNA sequence of NIPBL from the patient (upper) and a healthy individual (lower) showing a heterozygous 4-base deletion in exon 44 (c. 7423_7426delGACA) (black arrow).Venous blood samples were obtained from all the family members and 100 unrelated healthy individuals. Genomic DNA was extracted following a standard protocol. The coding exons and flanking intronic sequences of NIPBL (exons 2-47), SMC1A (exons 1-25) and SMC3 (exons 1-29) were amplified by PCR and bidirectional Sanger sequencing. This study was conducted in conformity with the Declaration of Helsinki and approved by Zhejiang University Review Board, and informed consent was given by all participating individuals. Sequencing analysis of the patient revealed a heterozygous, 4-base deletion (c.7423_7426delGACA) in exon 44 of NIPBL, which resulted in a frame-shift, leading to premature termination at codon 2504 (Figure 1C). The variant was not detected in the parents of our patient and the controls, suggesting a de novo event. No mutations in SMC1A or SMC3 were found. NIPBL is located at chromosome 5p13.1. It spans 189 kb of genomic DNA, and is composed of 47 exons. NIPBL is a novel protein displaying homology to Drosophila nipped-B and yeast sister chromatid cohesion protein 2 (scc2).4 The normal NIPBL molecule appears to play a role in sister chromatid cohesion and in regulating long-range enhancer-promoter interactions, possibly through interactions with the cohesin complex.5 To date, more than 80 causative mutations have been reported in patients with CdLS: 20% missense, 40% frame-shift, 20% nonsense, and 15% splice-site mutations (Human Genome Mutation Database: www.hgmd.cf.ac.uk). These mutations may cause loss-of-function alleles, and the severity of phenotypes generally correlates with the type of mutation.3 Analysis of genotype-phenotype associations demonstrated a trend toward a more severe phenotype in patients with nonsense or frame-shift mutations vs. patients with missense mutations or in-frame deletions, especially if the mutations occur in critical domains of the protein.3 In this study, we described the molecular genetic diagnosis of a Chinese girl with classic (severe) CdLS. She carried a heterozygous 4-based deletion (c.7423_7426delGACA) in NIPBL. According to the information available in the Human Genome Mutation Database and to the best of our knowledge, this is a novel de novo mutation of NIPBL that has not been reported elsewhere. The present case is consistent with previous genetic observations in other patients and provides further evidence that more severe symptoms are associated with more severe mutations, especially the frame-shift mutation type, in NIPBL. Because it is a heavy burden for society and families to raise these children, it is appropriate to offer genetic counseling to pedigrees which are affected or at risk. Generally, when the parents of an affected child are apparently clinically unaffected, the risk to the siblings of a child with CdLS has been estimated to be about 1.5% because of the possibility of germline mosaicism. Thus, the determination of genetic risk and discussion of the availability of prenatal testing should be provided before the next pregnancy. Our findings would facilitate family counseling and more definitive predictive prognosis.

BACKGROUND:Mutations in the KRT6A or KRT16 gene cause pachyonychia congenita type 1 (PC-1), while mutations in KRT16 or KRT6C underlie focal palmoplantar keratoderma (FPPK). A new classification system of PC has been adopted based on the mutated gene. PC rarely presents the symptoms of diffuse plantar keratoderma. Mutation in the tail domain of keratins is rarely reported. PC combined with fissured tongue has never been described.OBJECTIVES:To investigate the genotype-phenotype correlations between clinical features and gene mutational sites in two unrelated southern Chinese PC pedigrees (one family presented with specific fissured tongue, the other with diffuse plantar keratoderma).MATERIALS & METHODS:The whole coding regions of the KRT6A/KRT16/KRT17/KRT6B genes were amplified and directly sequenced to detect the mutation. To confirm the effect of the IVS8-2A>C mutation in KRT6A at the mRNA level, total RNA from the plantar lesion of a patient was extracted and reverse-transcribed to cDNA for sequence analysis.RESULTS:Two novel de novo mutations, a splice acceptor site variant IVS8-2A>C (p.S487FfsX72) in KRT6A and a heterozygous substitution c.AA373_374GG (p.N125G) within exon 1 of KRT16, were found separately in the two PC families.CONCLUSION:Genotype-phenotype correlations among PC patients with codon-125 mutation in KRT16 were established, while the phenotypes caused by the IVS8-2A>C mutation in KRT6A need further studies to confirm the rare feature of fissured tongue.

BACKGROUND:Genetic screening for germline mutations in the RET proto-oncogene has been extensively exploited worldwide to optimize the diagnostic and clinical management of multiple endocrine neoplasia type 2 (MEN2) patients and their relatives. However, a distinct lag period exists not only in the recognition but also in the medical treatment of patients with MEN2. Here we present a comprehensive genetic and clinical analysis of MEN2 among Chinese families followed from 1975 to 2011. Our series comprises 36 index cases and 134 relatives from 11 independent families.METHODS:Genetic diagnosis was performed in all participants by direct sequencing all relevant RET exons. Thyroidectomy was performed in 50 patients with varying cervical neck dissection procedures. Patients with pheochromocytoma (PHEO) underwent specific surgery. Demographic, clinical profiles, mutation types, tumor histopathologic features, and follow-up records were systematically analyzed.RESULTS:The RET mutations p.C634Y (n=34), p.C634R (n=6), p.C618S (n=13), p.V292M/R67H/R982C (n=7), p.L790F (n=2), and p.C634Y/V292M/R67H/R982C (n=1) were confirmed in 31 index cases and then identified in 32 at-risk relatives (mutation carriers), with MEN2A as the most common clinical subtype. The overall penetrance of PHEO in patients with MEN2A was 46.7%. A total of 50 patients underwent thyroidectomy, and there was a significant lowering of their mean age at thyroidectomy and the tumor diameter of the mutation carriers that were detected and operated on compared with the index cases (age at first surgery: 29.3 vs. 39.3 years, p<0.05; maximum size: 1.1 vs. 3.3 cm, p<0.001). There was also a decrease in the TNM staging and the proportion of patients who underwent inappropriate initial thyroid surgery (pN1: 31.6% vs. 100%, p<0.001; inappropriate surgery: 0% vs. 29%). Meanwhile, disease-free survival (DFS) increased (DFS: 100% vs. 58.1%, p<0.05). Both medullary thyroid carcinoma-specific (n=1) and PHEO-specific (n=5) deaths were reported during the study period.CONCLUSIONS:Our results further substantiate that gene scanning of all relevant RET exons is a powerful tool in the management of MEN2 patients, especially in asymptomatic carriers, and has led to earlier diagnosis and more complete initial treatment of patients with MEN2 in China.

AIM To investigate mitochondrial factors associated with Leber hereditary optic neuropathy (LHON) through complete sequencing and analysis of the mitochondrial genome of Chinese patients with this disease. METHODS Two unrelated southern Chinese families with LHON and 10 matched healthy controls were recruited, and their entire mitochondrial DNA (mtDNA) was amplified and sequenced with the universal M13 primer. Then DNA sequence analysis and variation identification were perfomed by DNAssist and Chromas 2 software and compared with authoritative databases such as Mitomap. RESULTS Mutational analysis of mtDNA in these two Chinese pedigrees revealed one common LHON-associated mutation, G11778A (Arg→His), in the MT-ND4 gene. In addition, there were two secondary mutations in Pedigree 1: C3497T (Ala→Val), and C3571T (Leu→Phe) in the MT-ND1 gene, which have not been reported; and two secondary mutations occurred in Pedigree 2: A10398G (Thr→Ala) in the MT-ND3 gene, and T14502C (Ile→Val) in the MT-ND6 gene. Three polymorphisms, A73G, G94A and A263G in the mtDNA control region, were also found. CONCLUSION Our study confirmed that the known MT-ND4*G11778A mutation is the most significant cause of LHON. The C3497T and C3571T mutations in Pedigree 1 were also both at hot-spots of MT-ND1; they may affect the respiratory chain in coordination with the primary mutation G11778A. In Pedigree 2, the two secondary mutations A10398G of MT-ND3 and T14502C of MT-ND6 may influence mitochondrial respiratory complex I, leading to the mitochondrial respiratory chain dysfunction which results in optic atrophy together with G11778A. Therefore, not only the common primary LHON mutation is responsible for the visual atrophy, but other secondary mtDNA mutations should also be considered when giving genetic counseling.