Compound heterozygous CAPN3 variants identified in a family with limb-girdle muscular dystrophy recessive 1.

Limb-girdle muscular dystrophy recessive 1 (LGMDR1), a rare subtype of muscular dystrophy, is characterized by progressive muscle weakness and degeneration with a predominant presentation on the shoulder, pelvic and proximal limb muscles. Variants in calcium-activated neutral proteinase 3 (CAPN3), which encodes an enzyme, calpain 3, are considered the major cause of LGMDR1. The present study was conducted to identify the variants responsible for clinical symptoms in a Chinese patient with limb-girdle muscular dystrophies (LGMDs) and explore its genotype-phenotype associations. A series of clinical examinations were conducted, including blood tests and magnetic resonance imaging scans of the lower legs, electromyography and muscle biopsy on the proband diagnosed with muscular dystrophies. Genomic DNA was extracted from the peripheral blood of a three-person family with LGMDs and pathogenic variants detected by whole-exome sequencing (WES) were verified by Sanger sequencing. The WES of this patient revealed compound heterozygous variants in CAPN3, c.2120A>G/p.(Asp707Gly) in exon 20 and c.2201_2202delAT/p.(Tyr734*) in exon 21, which were inherited from his parents and absent from 200 control individuals of similar ethnic origin, indicating that these variants are the pathogenic triggers of the LGMDR1 phenotype. Notably, these CAPN3 sequence variants were related to LGMDR1 pathogenesis in this three-person family. The newly discovered c.2201_2202delAT/p.(Tyr734*) expands the current CAPN3 variant spectrum, improving the understanding of the conditions required to develop molecular diagnostic tools and for genetic counseling, particularly for families with a history of autosomal recessive LGMDs.

Prognostic roles of mRNA expression of notch receptors in non-small cell lung cancer.

Notch signalling is aberrantly activated in human non-small cell lung cancer (NSCLC). Nevertheless, the prognostic roles of mRNA expression of four Notch receptors in NSCLC patients remain elusive. In this report, we reported the prognostic roles of Notch receptors in a total of 1,926 NSCLC patients through "The Kaplan-Meier plotter" (KM plotter) database which is capable to assess the effect of 22,277 genes on survival of NSCLC patients. We found that mRNA high expression level of Notch1 was associated with better overall survival (OS) for all NSCLC patients, hazard ratio (HR) 0.78 (0.69-0.89), p=0.00019, better OS in adenocarcinoma (Ade) patients, HR 0.59 (0.46-0.75), p=1.5e-05, as well as in squamous cell carcinoma (SCC) patients, HR 0.78 (0.62-0.99), p=0.044. mRNA high expression levels of Notch2 and Notch3 were associated with worsen OS for all NSCLC patients, as well as in Ade, but not in SCC patients. mRNA high expression level of Notch4 was not found to be associated with to OS for all NSCLC patients. In addition, mRNA high expression levels of Notch2, Notch3, but Notch4 are significantly associated with the NSCLC patients who have different smoking status. Our results indicate that mRNA expression of Notch receptors may have distinct prognostic values in NSCLC patients. These results will benefit for developing tools to accurately predict the prognosis of NSCLC patients.

Association between Retinoic acid receptor-ß hypermethylation and NSCLC risk: a meta-analysis and literature review.

Emerging evidence indicates that Retinoic acid receptor-ß (RARß) is a tumor suppressor in many types of tumor. However, whether or not RARß is a risk factor and is correlated to clinicopathological characteristics of non-small cell lung cancer (NSCLC) remains unclear. In this report, we performed a meta-analysis to determine the effects of RARß hypermethylation on the incidence of NSCLC and clinicopathological characteristics in human NSCLC patients. Final valuation and analysis of 1780 cancer patients from 16 eligible studies was performed. RARß hypermethylation was found to be significantly higher in NSCLC than in normal lung tissue, the pooled OR from 7 studies including 646 NSCLC and 580 normal lung tissues, OR = 6.05, 95% CI = 3.56-10.25, p<0.00001. RARß hypermethylation was significantly higher in adenocarcinoma (AC) compared to squamous cell carcinoma (SCC), pooled OR is 0.68 (95% CI = 0.52-0.89, p = 0.005). RARß hypermethylation was also found to occur significantly higher in smoker (n = 232) than non-smoker (n = 213) (OR = 2.46, 95% CI = 1.54-3.93, p = 0.0002). Our results indicate that RARß hypermethylation correlates well with an increased risk in NSCLC patients. RARß geneinactivation caused by RARß methylation contributes the NSCLC tumorigenesis and may serve as a potential risk factor, diagnostic marker and drug target of NSCLC.

An early biomarker and potential therapeutic target of RUNX 3 hypermethylation in breast cancer, a system review and meta-analysis.

Runt-related transcription factor 3 (RUNX3) methylation plays an important role in the carcinogenesis of breast cancer (BC). However, the association between RUNX3 hypermethylation and significance of BC remains under investigation. The purpose of this study is to perform a meta-analysis and literature review to evaluate the clinicopathological significance of RUNX3 hypermethylation in BC. A comprehensive literature search was performed in Medline, Web of Science, EMBASE, Cochrane Library Database, CNKI and Google scholar. A total of 10 studies and 747 patients were included for the meta-analysis. Pooled odds ratios (ORs) with corresponding confidence intervals (CIs) were evaluated and summarized respectively. RUNX3 hypermethylation was significantly correlated with the risk of ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC), OR was 50.37, p < 0.00001 and 22.66, p < 0.00001 respectively. Interestingly, the frequency of RUNX3 hypermethylation increased in estrogen receptor (ER) positive BC, OR was 12.12, p = 0.005. High RUNX3 mRNA expression was strongly associated with better relapse-free survival (RFS) in BC patients. In summary, RUNX3 methylation could be a promising early biomarker for the diagnosis of BC. High RUNX3 mRNA expression is correlated to better RFS in BC patients. RUNX3 could be a potential therapeutic target for the development of personalized therapy.

Screening of HHEX Mutations in Chinese Children with Thyroid Dysgenesis.

OBJECTIVE: Congenital hypothyroidism (CH) is a frequent neonatal endocrine disease with an incidence of about 1:2500 worldwide. Although thyroid dysgenesis (TD) is the most frequent cause of CH cases, its pathogenesis remains unclear. The aim of this study was to screen the hematopoietically-expressedhomeobox gene (HHEX) mutations in Chinese children with TD. METHODS: Genomic deoxyribonucleic acid was extracted from peripheral blood leukocytes in 234 TD patients from Shandong Province. Mutations in all exons and nearby introns of HHEX were analyzed by direct sequencing after polymerase chain reaction amplification. RESULTS: Sequencing analysis of HHEX indicated that no causative mutations were present in the coding regionof the TD patients. However, a genetic variant (IVS2+ 127 G/T, 10.26%) was observed in the intron 2 in HHEX. CONCLUSION: Our results indicate that the frequency of HHEX mutation is very low and may not be the main causative factor in Chinese TD patients. However, these results need to be replicated using larger datasets collected from different populations.

A novel missense mutation (I26M) in DUOXA2 causing congenital goiter hypothyroidism impairs NADPH oxidase activity but not protein expression.

CONTEXT: Dual-oxidase maturation factor 2 (DUOXA2) is a component of the thyroid hydrogen peroxidase (H2O2) generator, which is crucial for hormone synthesis. Genetic defects in DUOXA2 lead to an impairment of the H2O2-generating system, in turn causing congenital hypothyroidism (CH) with goiter. CASE DESCRIPTION: Our study aimed to identify DUOXA2 mutations associated with Chinese congenital goiter hypothyroidism patients and to examine the molecular mechanism underlying the genotype-phenotype relationship. All exons and flanking sequences of DUOXA2 in 75 unrelated CH with goiter patients were amplified by PCR and directly sequenced. DUOXA2 and DUOX2 protein expression levels were detected by Western blotting, and nicotinamide adenine dinucleotide phosphate oxidase activity was determined by measuring H2O2 generation in HeLa cells. A novel heterozygous missense mutation, c.C78G (p.I26M), and a homozygous nonsense mutation, c.C738G (p.Y246X), in DUOXA2 were identified in CH patients with mild transient and mild permanent goiter, respectively. In vitro experiments showed that the mutant I26M protein expression levels did not differ from those of wild-type DUOXA2 but that mutant I26M resulted in a complete deficiency of H2O2 generation. CONCLUSIONS: We identified a novel DUOXA2 mutation (I26M) causing CH with goiter, which affected H2O2 generation but did not alter the protein expression levels, further confirming the essential role of DUOXA2 in thyroid hormone synthesis.

[Effects of light condition on structure and photosynthetic characteristics of leaves in 'Hanfu' apple].

By using microscope technique and gas exchange method, the effects of light conditions on structure and photosynthetic characteristics of the leaves were studied with potted and fielded 'Hanfu' apple as materials. The results showed that the palisade tissue, spongy tissue and the total leaf thickness of 'Hanfu' apple was declined under the weak light environment, the palisade tissue were declined by 34.5% (pot) and 25.0% (field), and the total leaf thickness were declined by 27.1% (pot) and 18.3% (field). The light compensation point (LCP) of the field shading leaves was lowest (30.8 +/- 1.3 micromol x m(-2) x s(-1)), the saturation point (LSP) in full light was 22.7% (pot) and 48.2% (field) higher than in shading, respectively. The adaptability of the potted 'Hanfu' apple leaves built under different light conditions had different resistivity after exposing to high light, the startup time of maximum photosynthetic rates (15.4 micromol x m(-2) x s(-1) in full light and 12.7 micromol x m(-2) x s(-1) in shading) were different, which was 23 min and 33 min, respectively. Long-time shading impacted on the quality and photosynthetic capacity of 'Hanfu' apple leaves.

[Effects of different organic matter mulching on water content, temperature, and available nutrients of apple orchard soil in a cold region].

The effects of different organic matter covers on soil physical-chemical properties were investigated in a 'Hanfu' apple orchard located in a cold region. Four treatments were applied (weed mulching, rice straw mulching, corn straw mulching, and crushed branches mulching), and physical-chemical properties, including orchard soil moisture and nutrient contents, were compared among treatment groups and between organic matter-treated and untreated plots. The results showed that soil water content increased in the plots treated with organic matter mulching, especially in the arid season. Cover with organic matter mulch slowed the rate of soil temperature increase in spring, which was harmful to the early growth of fruit trees. Organic matter mulching treatments decreased the peak temperature of orchard soil in the summer and increased the minimum soil temperature in the fall. pH was increased in soils treated with organic matter mulching, especially in the corn straw mulching treatment, which occurred as a response to alleviating soil acidification to achieve near-neutral soil conditions. The soil organic matter increased to varying extents among treatment groups, with the highest increase observed in the weed mulching treatment. Overall, mulching increased alkali-hydrolyzable nitrogen, available phosphorus, and available potassium in the soil, but the alkali-hydrolyzable nitrogen content in the rice straw mulching treatment was lower than that of the control.

[Effects of high temperature on leaf photosynthetic characteristics and photosystem II photochemical activity of kernel-used apricot].

In order to explore the photosynthetic adaption mechanisms of kernel-used apricot under high temperature stress, gas exchange technique and chlorophyll fluorescence transient technique (JIP-test) were adopted to study the leaf photosynthetic characteristics and photosystem II (PS II) photochemical activity of 4 year-old 'Chaoren' (Armeniaca vulgaris x sibirica) growing on Horqin sandy land at 25 degrees C, 30 degrees C, 40 degrees C, and 50 degrees C. Within a definite temperature range, and as the temperature increased, the 'Chaoren' could enhance its leaf photosynthetic pigments content and ratio to maintain the light absorption, transfer, and conversion, and thereby, to ensure the function of photosynthetic apparatus. However, when the temperature exceeded the physiological adjustment threshold of leaves, the chlorophyll began to be decomposed, net photosynthetic rate (Pn) declined obviously, and intercellular CO2 concentration (Ci) increased, indicating that the decline in photosynthesis was limited by mesophyll factor. At 40 degrees C, the density of PS II reaction centers per excited cross-section (RC/CS0) dropped distinctly; and at 50 degrees C, the K phase (Wk) and J phase (Vj) in the O-J-I-P chlorophyll fluorescence transients increased distinctly, indicating that high temperature damaged the oxygen-evolving complex (OEC), donor sides, and PS II reaction centers. In addition, the minimum chlorophyll fluorescence (F0) at 50 degrees C increased significantly by 1.26 times, compared with the control, and the maximum photochemical efficiency (Fv/Fm) and performance index (PI(ABS)) reduced to 37.9% and 10.3% of the control, respectively. High temperature injured the function of the donor and acceptor sides in the PS II of photosynthetic apparatus, leading to the decrease of photosynthetic efficiency, and being one of the main mechanisms for the damage of photosynthetic apparatus in kernel-used apricot leaves under high temperature stress.

Quantitative analysis of phenolic compounds in Chinese hawthorn (Crataegus spp.) fruits by high performance liquid chromatography-electrospray ionisation mass spectrometry.

Eleven major phenolic compounds (hyperoside, isoquercitrin, chlorogenic acid, ideain, epicatechin, two procyanidin (PA) dimers, three PA trimers and a PA dimer-hexoside) were quantified in the fruits of 22 cultivars/origins of three species of the Chinese hawthorn (Crataegus spp.) by HPLC-ESI-MS-SIR. Hyperoside (0.1-0.8mg/g dry mass [DM]), isoquercitrin (0.1-0.3mg/g DM), chlorogenic acid (0.2-1.6mg/g DM), epicatechin (0.9-11.7mg/g DM), PA B2 (0.7-12.4mg/g DM), PA dimer II (0.1-1.5mg/g DM), PA trimer I (0.1-2.7mg/g DM), PA trimer II (0.7-6.9mg/g DM), PA trimer III (0.01-1.2mg/g DM) and a PA dimer-hexoside (trace-1.1mg/g DM) were detected in all the samples. Ideain (0.0-0.7mg/g DM) was found in all the samples except Crataegus scabrifolia. Significant correlations between the contents of individual PA aglycons were observed (r>0.9, P<0.01). A strong correlation between flavonols was also shown (r=0.71, P<0.01). Fruits of Crataegus pinnatifida var. major had higher contents of PAs but lower contents of flavonols compared with Crataegus brettschneideri. The fruits of C. scabrifolia contained the highest level of PA dimer-hexoside, which was present in trace amounts in the fruits of C. pinnatifida.

Acids, sugars, and sugar alcohols in Chinese hawthorn (Crataegus spp.) fruits.

Acids, sugars, and sugar alcohols in the fruits of 22 cultivars/origins of three species of hawthorn (Crataegus spp.) were analyzed by gas chromatography and mass spectrometry. Citric acid (2.0-8.4 g/100 g dry mass [DM]), quinic acid (0.5-5.6 g/100 g DM), malic acid (0.3-1.1 g/100 g DM), fructose (5.5-18.4 g/100 g DM), glucose (5.3-16.6 g/100 g DM), sorbitol (3.0-15.7 g/100 g DM), and myo-inositol (0.1-0.3 g/100 g DM) were found in all the samples. Sucrose was present only in C. scabrifolia and three cultivars of C. pinnatifida var. major. C. scabrifolia differed from other species by its high content of quinic acid. The cultivars of C. pinnatifida var. major and C. brettschneideri had a higher content of total sugars and a higher sugar/acid ratio than the natural origins of C. pinnatifida and C. scabrifolia (P < 0.05). The hawthorn samples analyzed fell into two groups rich in sugars and acids respectively. This is the first report of the profiles of sugars and sugar alcohols and the occurrence of quinic acid in hawthorn fruits.

[Microbial flora in Cerasus sachalinensis rhizosphere].

By using selected culture media, the microbes in Cerasus sachalinensis rhizosphere were isolated, identified and classified, with their community structure and dynamic changes at different growth stages of C. sachalinensis studied. The bacteria isolated were belonged to 15 genera, among which, Bacillus, Pseudomonas and Flavobacterium were the dominant ones. Flavus and Albosporus were the two dominant genera in seven groups of Actinomyces, and Mucor, Aspergillus and Penicillium were the main genera of fungi. The microbial flora varied with C. sachalinensis growth stage, being the richest at defoliation stage and the least at budding stage.

[Community structure and its dynamics of phosphobacteria in Cerasus sachalinensis rhizosphere and nonrhizosphere].

By using selective culture media, the phosphobacteria in Cerasus sachalinensis rhizosphere and nonrhizosphere were isolated and identified, with their community structure and dynamics studied. The phosphobacteria isolated from three test substrates belonged to 13 genera, and Bacillus, Pseudomonas and Serratia were the main ones. The substrate added with slag was most suitable for the growth of phosphobacteria, on which, phosphobacteria had the highest population number but the lowest diversity index. There was a difference in the population number of phosphobacteria in rhizosphere and nonrhizosphere during the growth period of C. sachalinensis. In rhizosphere, phosphobacteria had the highest number (6 genera) in withhold-growing period of shoot, less in fast growing period of shoot and in defoliation period, and the least in germination period. The diversity index of phosphobacteria in rhizosphere also varied with the growth period of C. sachalinensis, which was in the order of fast growing period of shoot > defoliation period > withhold-growing period of shoot. In nonrhizosphere, the population number and diversity index of phosphobacteria had a decreasing trend with the growth of C. sachalinensis. All of these indicated that there was an obvious rhizosphere effect on phosphobacteria.