Optimization of a Novel Engineered Ecosystem Integrating Carbon, Nitrogen, Phosphorus, and Sulfur Biotransformation for Saline Wastewater Treatment Using an Interpretable Machine Learning Approach.

The denitrifying sulfur (S) conversion-associated enhanced biological phosphorus removal (DS-EBPR) process for treating saline wastewater is characterized by its unique microbial ecology that integrates carbon (C), nitrogen (N), phosphorus (P), and S biotransformation. However, operational instability arises due to the numerous parameters and intricates bacterial interactions. This study introduces a two-stage interpretable machine learning approach to predict S conversion-driven P removal efficiency and optimize DS-EBPR process. Stage one utilized the XGBoost regression model, achieving an R2 value of 0.948 for predicting sulfate reduction (SR) intensity from anaerobic parameters with feature engineering. Stage two involved the CatBoost classification and regression model integrating anoxic parameters with the predicted SR values for predicting P removal, reaching an accuracy of 94% and an R2 value of 0.93, respectively. This study identified key environmental factors, including SR intensity (20-45 mg S/L), influent P concentration (<9.0 mg P/L), mixed liquor volatile suspended solids (MLVSS)/mixed liquor suspended solids (MLSS) ratio (0.55-0.72), influent C/S ratio (0.5-1.0), anoxic reaction time (5-6 h), and MLSS concentration (>6.50 g/L). A user-friendly graphic interface was developed to facilitate easier optimization and control. This approach streamlines the determination of optimal conditions for enhancing P removal in the DS-EBPR process.

Electrogenerated singlet oxygen and reactive chlorine species enhancing volatile fatty acids production from co-fermentation of waste activated sludge and food waste: The key role of metal oxide coated electrodes.

Electrochemical pretreatment (EPT) has shown to be superior in improving acidogenic co-fermentation (Co-AF) of waste activated sludge (WAS) and food waste (FW) for volatile fatty acids (VFAs). However, the influence of EPT electrode materials on the production of electrogenerated oxidants (such as singlet oxygen (1O2) and reactive chlorine species (RCS)), as well as their effects on properties of electrodes, the microbial community structure and functional enzymes remain unclear. Therefore, this study investigated the effects of various metal oxide coated electrodes (i.e., Ti/PbO2, Ti/Ta2O5-IrO2, Ti/SnO2-RuO2, and Ti/IrO2-RuO2) on EPT and subsequent Co-AF of WAS-FW. The results showed that EPT with Ti/PbO2, Ti/Ta2O5-IrO2, Ti/SnO2-RuO2 and Ti/IrO2-RuO2 electrodes generated 165.3-848.2 mg Cl2/L of RCS and 5.643 × 1011-3.311 × 1012 spins/mm3 of 1O2, which significantly enhanced the solubilization and biodegradability of WAS-FW by 106.4 %-233.6 % and 177.3 %-481.8 %, respectively. Especially with Ti/Ta2O5-IrO2 as the electrode material, an appropriate residual RCS (2.0-10.4 mg Cl2/L) remained in Co-AF step, resulted in hydrolytic and acidogenic bacteria (e.g., Prevotella_7, accounting for 78.9 %) gradually become dominant rather than methanogens (e.g., Methanolinea and Methanothrix) due to their different tolerance to residual RCS. Meanwhile, the functional gene abundances of hydrolytic and acidogenic enzymes increased, while the methanogenic enzymes deceased. Consequently, this reactor produced the highest VFAs up to 545.5 ± 36.0 mg COD/g VS, which was 101.8 % higher than that of the Control (without EPT). Finally, the economic analysis and confirmatory experiments further proved the benefits of WAS-FW Co-AF with EPT.

Thiosulfate/FeCl3 pre-treatment enhances short-chain fatty acid production and mitigates H2S generation during anaerobic fermentation of waste activated sludge: Performance, microbial community and ecological analyses.

Anaerobic fermentation (AF) has been identified as a promising method of transforming waste activated sludge (WAS) into high-value products (e.g., short-chain fatty acids (SCFAs)). This study developed thiosulfate/FeCl3 pre-treatment and investigated the effects of different thiosulfate/FeCl3 ratios (S:Fe = 3:1, 3:2, 1:1, 3:4 and 3:5) on SCFA production and sulfur transformation during the AF of WAS. At a S:Fe ratio of 1:1, the maximal SCFA yield (933.3 mg COD/L) and efficient H2S removal (96.5 %) were obtained. S:Fe ratios ≤ 1:1 not only benefited hydrolysis and acidification but largely mitigated H2S generation. These results were supported by the enriched acidogens and reduced sulfur-reducing bacteria (SRB). Molecular ecological network analysis further revealed that the keystone taxon (g_Saccharimonadales) was found in S:Fe = 1:1, together with reductions in associations among methanogens, acidogens and SRB. This work provides a strategy for enhancing high-value product recovery from WAS and minimising H2S emissions.

The effect of dietary awareness on household food waste.

How to reduce household food waste has emerged as an important issue worldwide. Considering the potential endogeneity issue, the present study utilizes the treatment effects model to analyse the effect of dietary awareness on household food waste using data from China. The results showed that improving dietary awareness could significantly increase household food waste overall. However, this impact is heterogeneous among households of different characteristics. Improving dietary awareness leads to more food waste for households with young and old food decision-makers, but contributes to less food waste for households of middle-aged makers. Also, the positive effect of dietary awareness on household food waste weakens as income increases. These findings propose a new perspective to understand the heterogeneity in household food waste in the context of dietary awareness promotion.

Cross resistance, inheritance and fitness advantage of cyetpyrafen resistance in two-spotted spider mite, Tetranychus urticae.

Cyetpyrafen belonging to mitochondrial electron transport inhibitors of complex II (METI II) has been widely applied to manage pest mites in China. To investigate the adaption of Tetranychus urticae in the evolution of cyetpyrafen resistance, a study of cross resistance, mode of inheritance and fitness comparison of resistance using indoor cyetpyrafen resistant strain (resistance ratio, RR > 2, 000-fold) was executed. Cyet-R showed serious cross resistance to cyenopyrafen (>2500-fold) and cyflumetofen (~190-fold). The number of resistant genes was evaluated via chi-square (χ2) test and the concentration-response curve regarding goodness-of-fit between observed and the expected mortality. The LC50s of F1RS (Cyet-R♀ × Tu-YN♂) and F1SR (Tu-YN♀ × Cyet-R♂) were 3126.30 mg/L and 2743.97 mg/L, respectively, without significance, suggesting autosomal inheritance. The degree of dominance (D) values of F1RS and F1SR ranged between 0 and 1, revealing an incompletely dominant inheritance in the tested population of Tetranychus urticae. Plots of concentration-response data for the orthogonal backcross and reverse backcross progenies showed a significant deviation from the expected lines, pointing out a polygenic inheritance. Besides, lifetable analysis showed a fitness advantage of Cyet-R with a significantly decreased adult preadult period and significantly increased total fecundity. This study suggested that cyetpyrafen resistance against T. urticae was inherited as autosomal, incompletely dominant and multigenetic and characterized with serious cross resistance and fitness advantage. Therefore, rational application and preventive strategy should be considered to sustain the efficacy of cyetpyrafen against T. urticae.

The Temporal and Spatial Changes of Health Inequality in Rural China.

This article estimates the temporal and spatial changes of health inequality in rural China from 2010 to 2018. Based on a panel database of 29,616 rural residents, the Health Utility Index (HUI) and a spatial econometric model are used for analysis. The results show that, on the temporal dimension, the health inequality of rural China first expands and then deflates. On the spatial dimension, the health inequality gradually deflates from eastern to western China. Furthermore, from 2010 to 2018, the high and low-value areas constantly changed among different provinces. After decomposing the causes of health inequality, it is found that behind the health inequality is the difference of socioeconomic-related status. Moreover, narrowing the difference in socioeconomic-related status is the key to improving health inequality.

Coronary microvascular dysfunction: An important interpretation on the clinical significance of transient ischemic dilation of the left ventricle on myocardial perfusion imaging.

PURPOSE: To further investigate the clinical significance of transient ischemic dilation (TID) on myocardial perfusion imaging (MPI) by analyzing the effect of anisodamine hydrobromide (a drug that can effectively ameliorate microcirculation) on the patients with isolated TID and the findings of previous literatures. METHODS: Total 107 patients with isolated TID (TID value≥1.11) were randomly divided into group A (n = 36; intravenous administration of anisodamine hydrobromide), group N (n = 36; intravenous administration of isosorbide dinitrate), and group C (n = 35; intravenous administration of normal saline). MPI and treadmill exercise test (TET) were performed again after 14-day course of intervention. Pre- and post-intervention frequencies of symptom were recorded. RESULTS: In group A, after intervention of anisodamine hydrobromide, the summed stress score (SSS) and TID value on MPI significantly decreased than those before intervention (P < 0.001), the durations of exercise (DEs) and metabolic equivalents (METs) in TET notably ascended (P < 0.001), as well as the symptom remarkably improved. In group N and group C, there were no significant differences in SSS, TID value, DEs, METs, and frequencies of symptom between pre- and post-intervention (P > 0.05). No significant improvement of symptoms in group N before and after treatment. CONCLUSIONS: TID with perfusion defect may usually predict a possibility of severe and extensive coronary artery disease (CAD). An isolated TID should be considered as a likelihood of coronary microvascular dysfunction (CMD). TET and coronary CT angiography (cCTA) are extremely helpful for the antidiastole on CAD and CMD. The administration of anisodamine hydrobromide might be an optional treatment for the patients with isolated TID.

Treatment of dilated cardiomyopathy caused by coronary microvascular dysfunction with anisodamine: a report of 5 cases.

The management of dilated cardiomyopathy (DCM) is well established. However, a subset of patients does not have recovery from or have recurrences of left ventricular (LV) dysfunction despite receiving optimal medical therapy. Coronary microvascular dysfunction (CMD) can result from structural and functional abnormalities at the intramural and small coronary vessel level affecting coronary blood flow autoregulation and consequently leading to impaired coronary flow reserve. Dilated myocardial phenotype may be responsible for CMD in DCM. Anisodamine can exert a significant effect on relieving microvascular spasm, and improving and dredging the coronary microcirculation. However, whether CMD can be potentially improved with anisodamine to make DCM better remains incompletely understood.

miR-429-3p/LPIN1 Axis Promotes Chicken Abdominal Fat Deposition via PPARγ Pathway.

To explore the regulatory mechanism of abdominal fat deposition in broilers, 100-day-old Sanhuang chickens (n = 12) were divided into high-fat and low-fat groups, according to the abdominal fat ratio size. Total RNA isolated from the 12 abdominal fat tissues was used for miRNA and mRNA sequencing. Results of miRNA and mRNA sequencing revealed that miR-429-3p was highly expressed in high-fat chicken whereas LPIN1 expression was downregulated. Further, we determined that miR-429-3p promoted preadipocyte proliferation and differentiation, whereas LPIN1 exerted an opposite effect. Notably, we found that the miR-429-3p/LPIN1 axis facilitated PPARγ pathway activation, which is closely associated with the progression of adipogenesis. In conclusion, our results provide evidence that a novel miR-429-3p/LPIN1 axis is involved in the regulation of adipogenesis, which may have a guiding role in the improvement of breeding for abdominal fat traits in broiler chickens.

Medical system and nutrition improvement for the rural elderly.

BACKGROUND: Insufficient nutrition intake has negatively influenced the health of the elderly in rural China where the problem of population aging is serious. The present study aims to explore whether the medical system, called the New Rural Cooperative Medical System (NRCMS), can improve the rural elderly's nutrition intake and the mechanism behind it. METHODS: The difference in differences (DID) model and the propensity score matching-difference in differences (PSM-DID) model are both performed to investigate the impact of the medical system on nutrition improvement for the rural elderly. Two thousand seven hundred eighty rural elderly samples tracked in 2000 and 2006 from the China Health and Nutrition Survey are analyzed. Indices for the elderly's nutrition intake includes daily average intake of energy, fat, protein, and carbohydrate. RESULTS: The results show that participation in the NRCMS can significantly increase the rural elderly's total energy intake, carbohydrate intake, and protein intake by 206.688 kcal, 36.379 g, and 6.979 g, respectively. A more significant impact of the NRCMS on nutrition intake is observed in the central and near-western where economic development is lagging behind. Also, compared to people of 18-60 age group, such impact is statistically more significant in the elderly for the carbohydrate intake. CONCLUSIONS: The NRCMS can improve the rural elderly's nutrition intake in China. As the population ages rapidly in rural China, the present study provides recommendations on how to improve nutrition and health status of the elderly from the aspect of the medical system.

The Effect of Health on Labour Supply of Rural Elderly People in China-An Empirical Analysis Using CHARLS Data.

In China, due to decades of the 'one-child policy' and continuous rural-urban labour migration, real population aging in rural areas is increasing more quickly than in urban areas, and the labour inputs in agricultural production are becoming ever more dependent on the elderly. Using CHARLS data, we examine the effect of health on the labour supply of rural elderly people. We construct a latent health stock index (LHSI) to eliminate measurement bias and then use this one-period lagged LHSI and the Heckman two-stage and the Bourguignon-Fournier-Gurand two-stage method to deal with the simultaneous causality of health and labour decisions and sample selectivity in model estimation. The results show that, in the overall level, the labour force participation and work time of rural elderly people increase significantly with the improvement of health. These effects on the males are sharply greater than on the females and are enhanced with age. In the subdivided agricultural and non-agricultural labour supply, health improvement is positively related with labour force participation of rural elderly and brings an employment allocation from agricultural section to non-agricultural section, especially on the males. However, as the work time, these relations are insignificant and invariant with gender and age.

[Clinical Assessment of Chemosensitivity Test in Xeno-free Culture of Autologous 
Malignant Effusion Cells from Patients with Advanced Lung Adenocarcinoma].

BACKGROUND: A great individual differences to chemotherapeutic effects existed in the patient with advanced lung cancer. How to choose the optimum regimens to achieve the individuation and maximum effect of chemotherapy for lung cancer is worth exploring. The study was designed to examine the effect of ex vitro chemo-sensitivity assay in xeno-free culture of autologous malignant effusion cells from patients with advanced lung adenocarcinoma. METHODS: The 50 treatment-naive patients with lung adenocarcinoma complicated with malignant pleural or pericardial effusions were enrolled. Effusions of all cases had been controlled by closed drainage and 300 mL-500 mL of which were retained under sterile condition from 25 cases (Chemo-sensitivity group). Primary malignant effusion cells were isolated from autologous effusions of the patients. Then, xeno-free culture (average 11 days) were intervened with 8 chemotherapeutic drugs commonly used in clinical practice and were determined by CCK-8 assay. Optimum regimens were selected for chemotherapy based on the results of chemosensitivity test. As a contrast, chemotherapy regimens for the other 25 patients (Control group) were on the basis of physician's clinical experience. RESULTS: After four cycles of chemotherapy, in Chemo-sensitivity group, 17 (68.0%) cases were determined for partial response (PR), 5 (20.0%) cases for stable disease (SD), and the objective response rate (ORR) was 68.0%, the disease control rate (DCR) was 88.0%. Meanwhile, in Control group, 9 (36.0%) cases were determined for PR, 7 (28.0%) cases for SD, and, the ORR was 36.0%, the DCR was 64.0%. There were significant differences between the two groups in ORR and DCR (P<0.05). To the end of follow-up, there were 21 cases of death in Chemo-sensitivity group as well as 22 cases in Control group. The mean progression-free survival (PFS) in Chemo-sensitivity group and Control group respectively were 10.0 months and 5.8 months, and the mean overall survival (OS) in the two groups were 30.2 months and 21.2 months respectively. There were also significant differences between the two groups in PFS and OS (P<0.05). Furthermore, the adverse reactions in both groups were mild and controllable. CONCLUSIONS: Xeno-free culture of autologous malignant effusion cells from patients with advanced lung adenocarcinoma and ex vitro chemo-sensitivity assay are beneficial to the rational choices of chemotherapeutic agents used in patients with lung adenocarcinoma complicated with malignant effusions, which is a worthy trial in personalized cell culture for individualized cancer therapy and further studies.

TBX5 loss-of-function mutation contributes to atrial fibrillation and atypical Holt-Oram syndrome.

Previous genome-wide association studies have demonstrated that single nucleotide polymorphisms in T­box (TBX)5 are associated with increased susceptibility to atrial fibrillation (AF), and a recent study has causally linked a TBX5 mutation to atypical Holt-Oram syndrome and paroxysmal AF. However, the prevalence and spectrum of TBX5 mutations in patients with AF remain to be elucidated. In the present study, a cohort of 190 unrelated patients with idiopathic AF were prospectively recruited, with 400 unrelated healthy individuals recruited as controls. The coding exons and flanking introns of the TBX5 gene were sequenced in the participants. The functional characteristics of the mutant TBX5 were delineated in contrast with its wild­type counterpart using a dual­luciferase reporter assay system. As a result, a novel heterozygous TBX5 mutation, p.P132S, was identified in an index patient with AF, with a mutational prevalence of ~0.53%. Genetic analysis of the proband's family showed that the mutation co­segregated with AF, and was transmitted in an autosomal dominant pattern. The missense mutation was absent in the 800 control chromosomes, and the altered amino acid was completely evolutionarily conserved across species. Functional analyses revealed that the mutant TBX5 had significantly reduced transcriptional activity. Furthermore, the mutation markedly decreased the synergistic activation between TBX5 and NK2 homeobox 5, another transcription factor which has been causatively linked to AF. The present study was the first, to the best of our knowledge, to report on the association between a TBX5 loss­of­function mutation and increased susceptibility to AF. These results provide novel insight into the molecular mechanism underpinning AF, and have potential implications in the development of novel prophylactic and therapeutic strategies for AF, the most common form of sustained cardiac arrhythmia.

The Anti-Apoptotic Role of Berberine in Preimplantation Embryo In Vitro Development through Regulation of miRNA-21.

Traditional Chinese medicinal herbs containing berberine have been historically used to prevent miscarriage. Here, we investigated whether the anti-apoptotic effects of berberine on pre-implantation embryonic development are regulated by miRNA-21. Mouse pronuclear embryos were cultured in medium with or without berberine, and some were then microinjected with a miRNA-21 inhibitor. The in vitro developmental rates of 2- and 4-cell embryos and blastocysts, blastocyst cell numbers, apoptotic rates, and apoptotic cell numbers were measured in each group. Furthermore, we examined the transcription levels of miRNA-21 and its target genes (caspase-3, PTEN, and Bcl-2) and their translation levels. Comparisons were made with in vivo-developed and untreated embryos. We found that berberine significantly increased the developmental rates and cell numbers of mouse blastocysts and decreased apoptotic cell rates in vitro. Berberine also significantly increased miRNA-21 and Bcl-2 transcription levels and significantly decreased caspase-3 and PTEN transcription levels. In embryos treated with a miRNA-21 inhibitor, the results followed the opposite trend; PTEN and caspase-3 transcription levels increased significantly, while the transcription level of Bcl-2 decreased significantly. Additionally, berberine treatment significantly increased the Bcl-2 protein level and significantly decreased the caspase-3 and PTEN protein levels in blastocysts, but there were no significant differences observed in the levels of these proteins in 2- and 4-cell embryos. This study revealed that miRNA-21 is important for pre-implantation embryonic development, especially blastocyst development in vitro. Berberine elevates miRNA-21 expression, decreases PTEN and caspase-3 levels, increases Bcl-2 levels, and exerts anti-apoptotic and pro-growth effects.

A novel TBX5 loss-of-function mutation associated with sporadic dilated cardiomyopathy.

Dilated cardiomyopathy (DCM) represents the most prevalent form of primary cardiomyopathy, and is the most common reason for heart transplantation and a major cause of congestive heart failure. Aggregating evidence demonstrates that genetic defects are associated with DCM, and a great number of mutations in >50 genes have been linked to DCM. However, DCM is a genetically heterogeneous disorder and the genetic components underpinning DCM in a significant proportion of patients remain unknown. In the present study, the coding exons and flanking exon­intron boundaries of the T-Box 5 (TBX5) gene, which encodes a T­box transcription factor required for normal cardiac development, were sequenced in 146 unrelated patients with sporadic DCM. The functional characteristics of the mutant TBX5 were assayed in contrast to its wild­type counterpart by using a dual­luciferase reporter assay system. As a result, a novel heterozygous TBX5 mutation, p.A143T, was identified in a patient with sporadic DCM. The missense mutation, which was absent in 400 control chromosomes, altered the amino acid that was completely conserved evolutionarily among species. Biological analyses revealed that the A143T mutation of TBX5 was associated with significantly decreased transcriptional activity on the promoter of the target gene atrial natriuretic factor (ANF), when compared to its wild­type counterpart. Furthermore, the A143T mutation abolished the synergistic activation of the ANF promoter between TBX5 and GATA binding protein 4 (GATA4), another crucial transcriptional factor for heart development. To the best of our knowledge, this is the first report on the association of a TBX5 loss­of­function mutation with an enhanced susceptibility to sporadic DCM, providing novel insight into the molecular mechanisms of the pathogenesis of DCM and suggesting potential implications for the prenatal prophylaxis and personalized treatment of this commonest primary myocardial disease.

Mutational spectrum of the NKX2-5 gene in patients with lone atrial fibrillation.

Atrial fibrillation (AF) is the most common form of sustained cardiac arrhythmia in humans and is responsible for substantial morbidity and mortality worldwide. Emerging evidence indicates that abnormal cardiovascular development is involved in the pathogenesis of AF. In this study, the coding exons and splice sites of the NKX2-5 gene, which encodes a homeodomain-containing transcription factor essential for cardiovascular genesis, were sequenced in 146 unrelated patients with lone AF as well as the available relatives of the mutation carriers. A total of 700 unrelated ethnically matched healthy individuals used as controls were genotyped. The disease-causing potential of the identified NKX2-5 variations was predicted by MutationTaster and PolyPhen-2. The functional characteristics of the mutant NKX2-5 proteins were analyzed using a dual-luciferase reporter assay system. As a result, two heterozygous NKX2-5 mutations, including a previously reported p.E21Q and a novel p.T180A mutation, were identified in two families with AF transmitted in an autosomal dominant pattern. The mutations co-segregated with AF in the families with complete penetrance. The detected substitutions, which altered the amino acids highly conserved evolutionarily across species, were absent in 700 control individuals and were both predicted to be causative. Functional analyses demonstrated that the NKX2-5 mutants were associated with significantly decreased transcriptional activity compared with their wild-type counterpart. The findings expand the spectrum of NKX2-5 mutations linked to AF and provide additional evidence that dysfunctional NKX2-5 may confer vulnerability to AF, suggesting the potential benefit for the early prophylaxis and personalized treatment of AF.

Traditional chinese medicine tongxinluo improves cardiac function of rats with dilated cardiomyopathy.

The study aimed at testing the hypothesis that tongxinluo capsule might exert its cardioprotective effect by preventing ventricular remodeling and improving coronary microvascular function in a rat model of doxorubicin-induced dilated cardiomyopathy (DCM). Rats that survived DCM induction were randomly divided into three groups to be given 1.5 g·kg(-1)·day(-1) (TXL-H, n = 9) or 0.15 g·kg(-1)·day(-1) (TXL-L, n = 10) of tongxinluo, or normal saline at the same volume (DCM-C, n = 10) intragastrically. Age matched normal rats treated with normal saline were used as normal controls (NOR-C, n = 9). After four weeks of treatment, the DCM-C, TXL-H, and TXL-L groups exhibited significant cardiac dysfunction, left ventricular remodeling, and coronary microvascular dysfunction, compared with the NOR-C rats. However, myocardial functional parameters were significantly improved and microvascular density (MVD) increased in the TXL-H group compared with the DCM-C group (all P < 0.01). Left ventricular remodeling was prevented. There were close linear relationships between CVF and LVEF (r = -0.683, P < 0.05), MVD and LVEF (r = 0.895, P < 0.05), and MVD and CVF (r = -0.798, P < 0.05). It was indicated that high-dose tongxinluo effectively improved cardiac function in rat model of DCM.

GATA4 loss-of-function mutation underlies familial dilated cardiomyopathy.

The cardiac transcription factor GATA4 is essential for cardiac development, and mutations in this gene have been implicated in a wide variety of congenital heart diseases in both animal models and humans. However, whether mutated GATA4 predisposes to dilated cardiomyopathy (DCM) remains unknown. In this study, the whole coding region and splice junction sites of the GATA4 gene was sequenced in 110 unrelated patients with idiopathic DCM. The available relatives of the index patient harboring an identified mutation and 200 unrelated ethnically matched healthy individuals used as controls were genotyped. The functional effect of the mutant GATA4 was characterized in contrast to its wild-type counterpart using a luciferase reporter assay system. As a result, a novel heterozygous GATA4 mutation, p.C271S, was identified in a family with DCM inherited as an autosomal dominant trait, which co-segregated with DCM in the family with complete penetrance. The missense mutation was absent in 400 control chromosomes and the altered amino acid was completely conserved evolutionarily among species. Functional analysis demonstrated that the GATA4 mutant was associated with significantly decreased transcriptional activity and remarkably reduced synergistic activation between GATA4 and NKX2-5, another transcription factor crucial for cardiogenesis. The findings provide novel insight into the molecular mechanisms involved in the pathogenesis of DCM, suggesting the potential implications in the prenatal diagnosis and gene-specific treatment for this common form of myocardial disorder.