The nicotine demethylase CYP82E4 is essential for the formation of red dapples on flue-cured leaves of cherry-red tobacco.

Common flue-cured tobacco (Nicotiana tabacum L.) primarily accumulates nicotine, and its flue-cured leaves exhibit a lemon appearance. In contrast, a spontaneous cherry-red variant (CR60) primarily accumulates nornicotine, accompanied by distinctive red dapples on the cured leaves. In this study, suppression of conversion of nicotine to nornicotine by genome editing resulted in decreased nornicotine and N-acyl nornicotines (NacNNs), and the subsequent disappearance of red dapples in CR60. Conversely, overexpression of CYP82E4 increased nornicotine and NacNNs accumulation, inducing a red dapple phenotype in common tobacco. Notably, nicotine conversion triggered significant alterations in leaf total sugars, alkaloids, and nitrogens. Metabolome analyses using 1352 identified compounds indicated nicotine conversion dramatically affected the entire metabolic network and induced unique metabolic responses across diverse genetic backgrounds. Further WGCNA analysis revealed that nicotine conversion caused substantial contents variation of alkaloids, flavonoids and amino acids and derivatives in cured leaves. Overall, this research provides valuable insights into the mechanisms underlying red dapple formation in cherry-red tobacco, elucidating profound influence of nicotine conversion on entire metabolic network.

Efficacy and tolerability of oxcarbazepine in the treatment of focal epilepsy in neonates and infants under 3 months of age: A single-center retrospective analysis.

OBJECTIVE: The neonatal and infantile period is the age group with the highest incidence of epilepsy, in which gene variants in sodium and potassium channels are an important etiology, so the sodium channel blocker class of antiseizure medications may be effective in the treatment of early onset epilepsy. This study aimed to summarize the efficacy and tolerability of oxcarbazepine (OXC) in the treatment of focal epilepsy in neonates and infants under 3 months of age. METHODS: A retrospective analysis of children with focal epilepsy onset within 3 months of age and treated with OXC in a tertiary pediatric epilepsy center in China was conducted. The efficacy, tolerability and influencing factors of OXC were evaluated. RESULTS: A total of 50 patients were enrolled, with a median age of epilepsy onset of 11.5 (2, 42) days. There were 32 cases of early infantile developmental and epileptic encephalopathy, 10 cases of self-limited neonatal or neonatal-infantile epilepsy, and 8 cases of focal epilepsy that could not be classified as epileptic syndrome. The median age of application of OXC was 47 (31, 66) days. The median follow-up time was 16.5 (10, 25) months, with 7 deaths. Thirty-eight cases (76.0 %) were effective with OXC treatment, including 28 cases (56.0 %) achieved seizure freedom. Of the 34 cases whose pathogenesis involved genetic factors, 19 cases with sodium/ potassium channel gene variants had higher effective and seizure-free rates than those with other gene variants. The most common adverse event was transient hyponatremia. 2 cases had rash and 2 cases had abnormal electrocardiogram, 3 of which discontinued OXC. SIGNIFICANCE: This single-center retrospective study suggests that OXC is effective and tolerable for the treatment of focal epilepsy in neonates and infants under 3 months of age. The efficacy of OXC is better in patients with sodium/ potassium channel gene variants.

Effect and environmental behaviour of microplastics in soil.

Soil microplastic pollution is currently a worldwide concern. Microplastics are organic pollutants that are abundant in the natural environment, are persistent and difficult to degrade and may endanger human health while harming the environment. This article offers a bibliometric analysis of the environmental behaviour of microplastics in soils, as well as a thorough statistical analysis of research goals and trends in this field. We conducted a thorough search of all relevant literature from 2012 to 2022 in the Web of Science core database. The data analysis shows that, starting in 2012, there has been an upward trend in the number of articles about soil microplastic pollution. It can also be seen that China is relatively ahead of the curve in this area of research, followed by the United Kingdom and the United States. This article also systematically describes the research hotspots in this field. The results show that the current research on soil microplastics is mainly focused on their identification, enrichment and toxicity, whereas studies on the migration and transformation of soil microplastics and the mechanism of interaction with other pollutants are still lacking. Our results provide ideas and prospects for future research in this field.

Disaster process-based spatiotemporal characteristics of apricot frost in the warm temperate zone (WTZ), China.

Frost stress is a major environmental factor that limits apricot growth in the warm temperate zone (WTZ) of China, and is always triggered by extreme low temperature weather processes. In this study, the characteristics of the apricot frost processes f(D, Tcum), which were identified from historical disaster representation, were analyzed and apricot frost evaluation indicators were developed, thus facilitating the process-based assessment and spatiotemporal analysis of apricot frost processes. Periods of low temperature that persist for 1~2, 3, and ≥4 days (i.e., duration days, D) provide the initial identification indicator for light, moderate, and severe apricot frost. The threshold ranges for Tcum are 0~3.9, 9.2~12.0, and >16.2 for D values of 1~2, 3, and ≥4, respectively. The northwest of the WTZ is dominated by apricot frost, with approximately 80% of apricot frost being light, followed by moderate and severe. Regional apricot frost exhibited a significant decreasing trend over the last four decades. A total of 29.65% of stations, which were mainly located in the northwest and middle parts of the study region, detected an increasing trend in apricot frost. The results provide technical support for targeted apricot frost level detection, and the process-based spatiotemporal characteristics of apricot frost can provide basic information for the prevention and mitigation of apricot frost.

The Top 100 Cited Articles on Acupuncture in the Last 20 Years: A Bibliometric Analysis.

BACKGROUND: Acupuncture has gained increasing international attention in recent decades. The act of incorporating acupuncture treatment into the routine treatment of COVID-19 in China drove us to review the 100 most influential articles of the last 20 years to learn about the current status and trends of acupuncture. METHOD: Articles related to acupuncture from January 1, 2001, to July 4, 2022, were searched in the Clarivate Analytics Web of Science Core Collection database. The top 100 most cited publications were selected, and information was extracted. Software such as VOSviewer, GraphPad Prism, Scimago Graphica, and CiteSpace were used to visualize and analyze the extracted data. RESULT: The 100 most cited articles were identified, with an average of 218 citations (range: 131-625). The majority of the top 100 articles were from the USA (n = 53). The institution that published the most highly cited papers was Harvard University (n = 16). The most influential team was Klaus Linde's group. Pain was the top-ranked journal in terms of the number of publications. The largest clusters for co-occurrence keyword analysis focused on acupuncture and electroacupuncture analgesia and brain imaging responses to acupuncture stimulation via functional MRI. The two highest strength burst keywords were "randomized controlled trials" and "osteoarthritis," with "randomized controlled trials" being a consistent burst keyword from 2011 to the present. CONCLUSION: This study provides insight into articles of historical significance in the field of acupuncture through bibliometric analysis. These data should provide clinicians and researchers with insight into future directions related to acupuncture.Die Akupunktur hat in den vergangenen Jahrzehnten zunehmend die internationale Aufmerksamkeit auf sich gezogen. Die Einbeziehung der Akupunktur in die Routinebehandlung von COVID-19 in China hat uns dazu veranlasst, die 100 einflussreichsten Artikel der letzten 20 Jahre zu überprüfen, um etwas über den aktuellen Stand und die Trends in der Akupunktur zu erfahren.Artikel mit Bezug zu Akupunktur vom 1. Januar 2001 bis 4. Juli 2022 wurden in der Clarivate Analytics Web of Science Core Collection Datenbank gesucht. Die 100 am häufigsten zitierten Veröffentlichungen wurden ausgewählt und die Informationen wurden extrahiert. Visualisierung und Analyse der extrahierten Daten erfolgten mithilfe der Software VOSviewer, GraphPad Prism, Scimago Graphica und CiteSpace.Es wurden die 100 am häufigsten zitierten Artikel ermittelt mit durchschnittlich 218 Zitierungen (Spanne: 131 bis 625). Der Großteil der 100 meistzitierten Artikel stammte aus den Vereinigten Staaten (n = 53). Die Institution, die die meisten zitierten Artikel veröffentlichte, war die Harvard-Universität (n = 16). Das einflussreichste Team war die Gruppe von Klaus Linde. Was die Zahl der Veröffentlichungen betrifft, war Pain die am häufigsten zitierte Zeitschrift. Bei der Analyse der Koinzidenz von Schlüsselwörtern lag der Fokus der größten Cluster auf Akupunktur-und Elektroakupunktur-Analgesie sowie auf den Reaktionen auf Akupunkturstimulation in den bildgebenden Untersuchungen des Gehirns mittels funktioneller MRT. Die beiden stärksten Burst-Schlüsselwörter waren "randomisierte kontrollierte Studien" und "Osteoarthritis," wobei "randomisierte kontrollierte Studien" von 2011 bis heute durchgängig ein Burst-Schlüsselwort ist.Diese Studie bietet einen Einblick in historisch bedeutsame Artikel auf dem Gebiet der Akupunktur mittels bibliometrischer analyse. Diese Daten sollen Klinikern und Forschern einen Einblick in zukünftige Richtungen im Zusammenhang mit Akupunktur geben.

The clinical spectrum associated with ATP1A2 variants in Chinese pediatric patients.

PURPOSE: To evaluate the clinical spectrum associated with ATP1A2 variants in Chinese children with hemiplegia, migraines, encephalopathy or seizures. METHODS: Sixteen children (12 males and 4 females), including ten patients with ATP1A2 variants whose cases had been published previously, were identified using next-generation sequencing. RESULTS: Fifteen patients had FHM2 (familial hemiplegic migraine type 2), including three who had AHC (alternating hemiplegia of childhood) and one who had drug-resistant focal epilepsy. Thirteen patients had DD (developmental delay). The onset of febrile seizures, which occurred between 5 months and 2 years 5 months (median 1 year 3 months) was earlier than the onset of HM (hemiplegic migraine), which occurred between 1 year 5 months and 13 years (median 3 years 11 months). Disturbance of consciousness subsided first, at 40 h to 9 days (median 4.5 days); hemiplegia and aphasia were resolved slowly, taking 30 min to 6 months (median 17.5 days) for the former and 24 h to over 1 year (median 14.5 days) for the latter. Cranial MRI showed edema in the cerebral hemispheres, mainly the left hemisphereacute attacks. All thirteen FHM2 patients recovered to baseline in 30 min to 6 months. Fifteen patients had between 1 and 7 (median 2) total attacks between the baseline and follow-up timepoints. We report twelve missense variants, including a novel variant ATP1A2 variant, p.G855E. CONCLUSIONS: The known genotypic and phenotypic spectra of Chinese patients with ATP1A2-related disorders were further expanded. Recurrent febrile seizures and DD combined with paroxysmal hemiplegia and encephalopathy should raise the clinical suspicion of FHM2. The avoidance of triggers and thus the prevention of attacks may be the most effective therapy for FHM2.

Characterization of feruloyl esterases from Pecoramyces sp. F1 and the synergistic effect in biomass degradation.

Feruloyl esterase (FAE; EC 3.1.1.73) cleaves the ester bond between ferulic acid (FA) and sugar, to assist the release of FAs and degradation of plant cell walls. In this study, two FAEs (Fae13961 and Fae16537) from the anaerobic fungus Pecoramyces sp. F1 were heterologously expressed in Pichia pastoris (P. pastoris). Compared with Fae16537, Fae13961 had higher catalytic efficiency. The optimum temperature and pH of both the FAEs were 45 ℃ and 7.0, respectively. They showed good stability-Fae16537 retained up to 80% activity after incubation at 37 ℃ for 24 h. The FAEs activity was enhanced by Ca2+ and reduced by Zn2+, Mn2+, Fe2+ and Fe3+. Additionally, the effect of FAEs on the hydrolytic efficiency of xylanase and cellulase was also determined. The FAE Fae13961 had synergistic effect with xylanase and it promoted the degradation of xylan substrates by xylanase, but it did not affect the degradation of cellulose substrates by cellulase. When Fae13961 was added in a mixture of xylanase and cellulase to degrade complex agricultural biomass, it significantly enhanced the mixture's ability to disintegrate complex substrates. These FAEs could serve as superior auxiliary enzymes for other lignocellulosic enzymes in the process of degradation of agricultural residues for industrial applications.

Synthesis of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one from maltol and its taste identification.

2,3-Dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) exists in many foods, and its effect on taste is controversial. The aim of this study was to clarify whether DDMP has bitter taste or not. For this purpose, DDMP was synthesized from maltol instead of from glucose for the first time. In contrast, DDMP derived from glucose was also prepared and further purified. Their structures were identified by NMR and MS, and considered to be the same substance. The sensory analysis showed that DDMP derived from maltol was tasteless. Further studies indicated that some impurities in Maillard reaction made DDMP derived from glucose taste bitter.

Interactions between Anaerobic Fungi and Methanogens in the Rumen and Their Biotechnological Potential in Biogas Production from Lignocellulosic Materials.

Anaerobic fungi in the digestive tract of herbivores are one of the critical types of fiber-degrading microorganisms present in the rumen. They degrade lignocellulosic materials using unique rhizoid structures and a diverse range of fiber-degrading enzymes, producing metabolic products such as H2/CO2, formate, lactate, acetate, and ethanol. Methanogens in the rumen utilize some of these products (e.g., H2 and formate) to produce methane. An investigation of the interactions between anaerobic fungi and methanogens is helpful as it provides valuable insight into the microbial interactions within the rumen. During the last few decades, research has demonstrated that anaerobic fungi stimulate the growth of methanogens and maintain methanogenic diversity. Meanwhile, methanogens increase the fiber-degrading capability of anaerobic fungi and stimulate metabolic pathways in the fungal hydrogenosome. The ability of co-cultures of anaerobic fungi and methanogens to degrade fiber and produce methane could potentially be a valuable method for the degradation of lignocellulosic materials and methane production.

Mahalanobis distance based similarity regression learning of NIRS for quality assurance of tobacco product with different variable selection methods.

Quality assurance is one of the key issues in tobacco industry and many efforts have been put on the quality control. This paper introduces a new chemometrics technique to estimate the "quality similarity rate", which is used for quality control. The value of the quality similarity rate represents the similarity degree between the products and the standard reference samples, which is a global parameter that can be generated by either human assessors or machine learning. Supervised similarity regression models are built to automatically estimate the quality similarity rate value from NIRS data of tobacco leaf and smoke. For the similarity regression learning, the metric matrix is generated by a novel method which calculates the Mahalanobis distance from the segmented near infrared spectroscopy (NIRS). The results show the similarity regression learning can predict the quality similarity score well in high speed and can be improved with lasso (least absolute shrinkage and selection operator) related feature selection algorithms such as sRDA (sparse redundancy analysis) and glmnet.

Effect of hydroxyl on antioxidant properties of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one to scavenge free radicals.

It is well known that 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) is usually formed in the Maillard reaction and it contributes to the antioxidant properties of Maillard reaction intermediates. A series of hydroxyl group protected DDMP derivatives were synthesized to further understand the source of antioxidant activity. Antioxidant abilities of the DDMP derivatives were evaluated by scavenging the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS˙+), 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), and galvinoxyl radical, respectively. It was found that the introduction of protecting groups to the free hydroxyl groups of DDMP decreases their reducing abilities. In particular, the hydroxyl group at the olefin position exhibited a remarkable impact on the antioxidant activity of DDMP, indicating that the unstable enol structure in the DDMP moiety is the key factor for its antioxidant activity.

Elevated TPOAb is a Strong Predictor of Autoimmune Development in Patients of Type 2 Diabetes Mellitus and Non-Alcoholic Fatty Liver Disease: A Case-Control Study.

OBJECTIVE: The aim of this study was to assess the prevalence of thyroid autoimmunity in T2DM with NAFLD, furthermore explore the relationship between elevated TPOAb titer and the severity of NAFLD. METHODS: A total of 400 patients with T2DM were divided into two groups according to NAFLD. Thyroid function and other metabolic indicators were measured. RESULTS: There were more TPOAb-positive patients in both groups, and the prevalence of TPOAb positive was significantly different in two groups (17% vs 6.9%, p< 0.01). FT4 was significantly lower in patients with T2DM with NAFLD (median FT4 0.89 vs 1.08, p < 0.001), while TSH was increased (median TSH 2.85 vs 2.28, p < 0.05). In patients with T2DM with NAFLD, the proportion of women in the thyroid autoimmune-positive group was significantly higher than the negative (71.1% vs 46%, p < 0.01). Similarly, thyroid autoimmune-positive T2DM and NAFLD patients had lower FT4 levels (median FT4 0.59 vs 0.92, p < 0.001), higher TSH levels (median TSH 3.65 vs 2.67, p < 0.001), and much higher TPOAb/TGAb (median TPOAb/TGAb 6.8 vs 1.46, p < 0.001). The increase of TPOAb was significantly correlated with the severity of fatty liver. HbA1c, TC, TG, TSH, TPOAb/TGAb and severity of fatty liver were risk factors of thyroid autoimmunity. CONCLUSION: Autoimmune thyroid disease is more common in patients with T2DM complicated with NAFLD. Elevated TPOAb titer is closely related to fatty liver, suggesting that elevated TPOAb titer is a predictor of autoimmune development in T2DM with NAFLD.

Clinical characteristics and gene mutation analysis of an adult patient with ETFDH­related multiple acyl­CoA dehydrogenase deficiency.

Multiple acyl­CoA dehydrogenase deficiency (MADD) is a rare autosomal recessive disorder of fatty acid metabolism caused by defects in electron transfer flavoprotein (ETF) or electron transfer flavoprotein dehydrogenase (ETFDH). These defects are mainly classified into the neonatal and late­onset types, based on their clinical manifestations. ETFDH gene mutations are generally considered to be associated with the late­onset type. The present study reported an adult woman with late­onset MADD accompanied with biochemical and muscle biopsy findings indicating metabolic disorders. Gene sequencing analysis showed that the c.1514T>C homozygous mutation in the region of the 12th exon of the ETFDH gene, which led to the amino acid substitution p.I505T (isoleucine > threonine), resulting in defective ETFDH protein function. The results of family verification revealed that the homozygous mutation originated from her parents. The female patient was treated with a large dose of vitamin B2, L­carnitine and coenzyme Q10, and the symptoms were significantly relieved. The c.1514T>C mutation in the ETFDH gene, was considered as a novel pathogenic mutation that had not been previously reported. Therefore, it was hypothesized that this mutation was responsible for the clinical characteristics of the adult female patient. Overall, this novel mutation could expand the spectrum of the ETFDH gene mutation and provide the basis for the etiological and prenatal diagnosis of MADD.

Analysis of angiographic characteristics and intervention of vitamin D in type 2 diabetes mellitus complicated with lower extremity arterial disease.

AIMS: Previous studies have suggested that type 2 diabetes mellitus with lower extremity arterial disease is related to 25-hydroxyvitamin D deficiency. The purpose of this study is to explore the relation between vitamin D supplementation and the characteristics of type 2 diabetes mellitus complicated with lower extremity arterial disease. METHODS: The clinical data of 514 patients and 148 healthy subjects treated in the First Hospital of Lanzhou University from January 2012 to June 2019 were collected, including the clinical data, ankle-brachial index, and medical records of lower limb artery angiography. We divided the patients into control group (NC group), type 2 diabetes mellitus group (DM group), lower extremity artery disease in type 2 diabetes mellitus without vitamin D supplement group (DM1 group) and lower extremity artery disease in type 2 diabetes mellitus with vitamin D supplement group (DM2 group). The level of serum 25(OH)D was analyzed and the characteristics of arterial lesions of lower extremities were compared by DSA arteriography in DM1 and DM2 group, respectively. RESULTS: Compared with the NC group, serum 25(OH)D level decreased in DM group (25.39 ± 4.94 ng/mL vs 19.43 ± 5.98 ng/mL) and significantly decreased in DM1 and DM2 group (14.22 ± 5.64 ng/mL vs 17.36 ± 6.25 ng/mL). However, the level of serum 25(OH)D in the DM2 group was higher than that in the DM1 group. Compared with the DM1 group, the disease rate of the inferior knee artery (65% vs 39.3%) and occlusion rate (11.5% vs 3.7%)were decreased in the DM2 group (P < 0.05). Logistic stepwise regression analysis showed that serum 25(OH)D level was a risk factor for lower extremity arterial disease in patients with type 2 diabetes mellitus (OR = 0.898,95%CI = 0.856-0.942). CONCLUSIONS: The serum level of 25(OH)D in patients with type 2 diabetes mellitus complicated with lower extremity arterial disease is decreased, and level of 25 (OH) D is related to stenosis and occlusion rate, especially in inferior genicular artery in T2DM complicated with LEAD. A high level of 25(OH)D may be a protective factor in type 2 diabetes with lower extremity arterial disease.

Association between diabetic neuropathy and osteoporosis in patients: a systematic review and meta-analysis.

OBJECTIVES: Many studies have explored the association between neuropathy and osteoporosis in patients with diabetes mellitus. However, the results still remain inconsistent and controversial. We aimed to estimate the association between diabetic neuropathy and osteoporosis. METHODS: Databases, including PubMed, Embase, Web of Science, the Cochrane library, Chinese Biomedical Literature Database (CBM), and Wanfang, were screened from inception to 30 March 2020. Studies were selected and data were extracted by two independent reviewers. Study characteristics and quality sections were reviewed independently. Pooled ORs and 95% CIs were calculated using random effects model when evidence of heterogeneity was present; otherwise, fixed effects model was used. Meta-regression and subgroup analyses were performed to explore the source of heterogeneity. Sensitivity analysis and publication bias were also tested. RESULTS: A total of 11 studies with 27,585 participants were included in this analysis which indicated that there was an increased odd between diabetic neuropathy and osteoporosis (overall OR 2.20, 95% CI 1.71-2.83). In the subgroup analyses and meta-regression, diabetic neuropathy has no significant difference in osteoporosis or fracture (p = 0.532). And osteoporosis also has no significant difference in type 1 or type 2 diabetic neuropathy (p = 0.668). CONCLUSIONS: This meta-analysis suggests that patients with diabetic neuropathy have a significantly increased chance of developing osteoporosis, even fragility fracture. The clinicians should pay more attention to the patients with diabetic neuropathy. Further studies were still needed to explore the confounding factors among studies and to elucidate the underlying biological mechanisms.

Effect of thyrotropin suppressive therapy on heart rate variability and QT dispersion in patients with differentiated thyroid cancer.

The effects of thyrotropin (TSH) suppressive therapy on autonomic regulation and ventricular repolarization in patients with differentiated thyroid cancer (DTC) have not been elucidated. The aim of present study was to evaluate variation in heart rate variability (HRV) and QT dispersion after TSH suppressive therapy in patients with DTC.Cases, defined as 271 patients with DTC within 1 year of exogenous levothyroxine, and all patients underwent a full history, physical examination, including standard 12 lead electrocardiogram (ECG), and 24 h ambulatory ECG monitoring (Holter) with normal free thyroxine (FT4) and free triiodothyronine (FT3) with levothyroxine. To evaluate effects of TSH suppressive therapy on HRV and QT dispersion, patients were divided into three groups according to different levels of TSH: TSH < 0.1 mIU/L group and 0.1 ≤ TSH < 0.5 mIU/L group were as TSH suppression groups, and 0.5 ≤ TSH < 2.0 mIU/L group was as TSH replacement group.Comparing with 0.5 ≤ TSH < 2.0 mIU/L group, significant changes in both time and frequency domain of HRV and QT dispersion were observed in TSH < 0.1 mIU/L group (P < .001: SDNN, SDANN, HF, LF/HF, QTd, and QTcd; P < .05: rMSSD) and 0.1 ≤ TSH < 0.5 mIU/L group (P < .001: SDNN, HF, LF/HF, QTd, and QTcd), and especially were more pronounced in TSH < 0.1 mIU/L group. Moreover, we found that TSH level was proportional to SDNN (ß = 15.829, P < .001), but inversely proportional to LF/HF (ß = -0.671, P < .001), QTd (ß = -16.674, P < .001) and QTcd (ß = -18.314, P < .001) in DTC patients with exogenous levothyroxine.Compared with euthyroid state, patients with suppressed serum TSH have increased sympathetic activity in the presence of diminished vagal tone, ultimately showed sympathovagal imbalance and with an increased inhomogeneity of ventricular recovery times. These findings revealed that TSH suppression therapy had a significant impact on cardiovascular system and had certain guiding role in the treatment and management of patients with DTC.

The enrichment of anaerobic fungi and methanogens showed higher lignocellulose degrading and methane producing ability than that of bacteria and methanogens.

In this study, rumen content was used to obtain three enrichments of anaerobic fungi and methanogens (F + M enrichment), bacteria and methanogens (B + M enrichment), and whole rumen content (WRC enrichment), to evaluate their respective ability to degrade lignocellulose and produce methane. Among the treatments, F + M enrichment elicited the strongest lignocellulose degradation and methane production ability with both rice straw and wheat straw as substrates. Quantitative real-time PCR analysis and diversity analyses of methanogens in the three enrichment treatments demonstrated that F + M had larger number of 16S rRNA gene copies of methanogens and higher relative abundance of Methanobrevibacter, the predominant methanogen found in all enrichments. Caecomyces was the main anaerobic fungal genus for co-culturing to provide substrates for methanogens in this enrichment. Importantly, the F + M enrichment was stable and could be maintained with transfers supplied every 3 days, confirming its potential utility in anaerobic digestion for lignocellulose degradation and methane production.

Temporal changes in vegetation around a shale gas development area in a subtropical karst region in southwestern China.

Over the past decade, various aspects of China's fragile karst environments, including net primary productivity (NPP), have been changed or threatened by shale gas development. This industry is still developing, so it is important to understand what drives environmental changes, particularly in NPP, when shale gas pads are constructed in sensitive areas. Few previous studies have addressed this issue, so we quantified how the NPP changed, and what drove the changes, when a large shale gas area was developed at the end of 2012 in a mountainous karst area in Sichuan Province. We calculated the trend in the normalized difference vegetation index (NDVI) from 2012 to 2017 and used the Carnegie-Ames-Stanford Approach (CASA) model to calculate the changes in NPP at different distances from the pads using remote sensing images for July 2012 and July 2017 and field survey data from July 2017. We then identified the factors that drove the changes with Geodetector. The results showed that the NDVI increased across 64.2% of the shale gas development area from 2012 to 2017 because of climate change, and only showed a significant decrease across 0.3% of the area, mainly because of the shale gas development. The NPP decreased by 110.1 t because of the shale gas development in July 2017, or by about 0.35% of the total NPP. Of this, 93.8 t were associated with the pad construction areas, and 16.3 t were associated with the area around the pads. The changes in NPP around the shale gas pads were mainly confined to within 150 m during the construction phase and 90 m once the construction was completed. The NPP at different distances from the pads during the construction period was related to the distance from the pad, slope, and land use. Once completed, the NPP mainly varied with distance, land use, and the distance from the pad to rural settlements. The NPP was most strongly influenced by the distance from the pad and the area of the pad. We suggest that, when planning the construction of shale gas pads, the pads should be sited on gently sloping areas, the number of wells on each pad should be optimized, land use type changes outside the pad should be limited, and the land beyond the pads should be reclaimed in a timely manner to allow the NPP to recover.

Diagnosis of intellectual disability/global developmental delay via genetic analysis in a central region of China.

BACKGROUND: Advanced technology has become a valuable tool in etiological studies of intellectual disability/global developmental delay (ID/GDD). The present study investigated the role of genetic analysis to confirm the etiology in ID/GDD patients where the cause of the disease was uncertain in central China. METHODS: We evaluated 1051 ID/GDD children aged 6 months to 18 years from March 2009 to April 2017. Data concerning basic clinical manifestations were collected, and the method of etiology confirmation was recorded. Genome-wide copy number variations (CNVs) detection and high-throughput sequencing of exons in the targeted regions was performed to identify genetically-based etiologies. We compared the incidence of different methods used to confirm ID/GDD etiology among groups with differing degrees of ID/GDD using the Chi-square or Fisher exact probability test. RESULTS: We recruited 1051 children with mild (367, 34.9%), moderate (301, 28.6%), severe (310, 29.5%), and profoundly severe (73, 6.9%) ID/GDD. The main causes of ID/GDD in the children assessed were perinatal factors, such as acquired brain injury, as well as single gene imbalance and chromosomal gene mutation. We identified karyotype and/or CNVs variation in 46/96 (47.9%) of cases in severe ID/GDD patients, which was significantly higher than those with mild and moderate ID/GDD of 34/96 (35.4%) and 15/96 (15.6%), respectively. A total of 331/536 (61.8%) patients with clear etiology have undergone genetic analysis while 262/515 (50.9%) patients with unclear etiology have undergone genetic analysis (χ = 12.645, P < 0.001). Gene structure variation via karyotype analysis and CNV detection increased the proportion of children with confirmed etiology from 51.0% to 56.3%, and second-generation high-throughput sequencing dramatically increased this to 78.9%. Ten novel mutations were detected, recessive mutations in X-linked genes (ATPase copper transporting alpha and bromodomain and WD repeat domain containing 3) and dominant de novo heterozygous mutations in X-linked genes (cyclin-dependent kinase like 5, protocadherin 19, IQ motif and Sec7 domain 2, and methyl-CpG binding protein 2) were reported in the study. CONCLUSIONS: The present study indicates that genetic analysis is an effective method to increase the proportion of confirmed etiology in ID/GDD children and is highly recommended, especially in ID/GDD children with uncertain etiology.