[Clinical phenotype and genetic analysis of six Chinese patients affected with Acromicric dysplasia due to variants of FBN1 gene].

OBJECTIVE: To retrospectively analyze the clinical and genetic characteristics of six patients with Acromicric dysplasia due to variants of the FBN1 gene. METHODS: Six patients who had visited the Affiliated Hospital of Qingdao University between February 2018 and October 2020 were selected as the study subjects. Clinical data of the patients were collected. High-throughput sequencing was carried out. And candidate variants were verified by Sanger sequencing. RESULTS: All of the six patients had presented with severe short stature (< 3s), brachydactyly, short and broad hands and feet. Other manifestations included joint stiffness, facial dysmorphism, delayed bone age, liver enlargement, coracoid femoral head, and lumbar lordosis. Genetic testing revealed that all had harbored heterozygous variants of the FBN1 gene. Patient 1 had harbored a c.5183C>T (p.A1728V) missense variant in exon 42, which had derived from his father (patient 2). Patient 3 had harbored a c.5284G>A (p.G1762S) missense variant in exon 43, which had derived from her mother (patient 4). Patient 5 had harbored a c.5156G>T (p.C1719F) missense variant in exon 42, which was de novo in origin. Patient 6 had harbored a c.5272G>T (p.D1758Y) missense variant in exon 43, which was also de novo in origin. The variants carried by patients 1, 3 and 6 were known to be pathogenic. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the FBN1: c.5156G>T was rated as a pathogenic variant (PS2+PM1+PM2_Supporting +PM5+PP3). CONCLUSION: All of the six patients had severe short stature and a variety of other clinical manifestations, which may be attributed to the variants of the FBN1 gene.

Alterations in the gut microbiota of toddlers with cow milk protein allergy treated with a partially hydrolyzed formula containing synbiotics: A nonrandomized controlled interventional study.

Formulas containing intact cow milk protein are appropriate alternatives when human milk (HM) is not feasible. However, for babies with a physician-diagnosed cow milk protein allergy (CMPA), hydrolyzed formulas are needed. We conducted a 3-month, open-label, nonrandomized concurrent controlled trial (ChiCTR2100046909) between June 2021 and October 2022 in Qingdao City, China. In this study, CMPA toddlers were fed with a partially hydrolyzed formula containing synbiotics (pHF, n = 43) and compared with healthy toddlers fed a regular intact protein formula (IF, n = 45) or HM (n = 21). The primary endpoint was weight gain; the secondary endpoints were changes in body length and head circumference of both CMPA and healthy toddlers after 3-month feeding; and the exploratory outcomes were changes in gut microbiota composition. After 3 months, there were no significant group differences for length-for-age, weight-for-age, or head circumference-for-age Z scores. In the gut microbiota, pHF feeding increased its richness and diversity, similar to those of IF-fed and HM-fed healthy toddlers. Compared with healthy toddlers, the toddlers with CMPA showed an increased abundance of phylum Bacteroidota, Firmicutes, class Clostridia, and Bacteroidia, and a decreased abundance of class Negativicutes, while pHF feeding partly eliminated these original differences. Moreover, pHF feeding increased the abundance of short-chain fatty acid producers. Our data suggested that this pHF partly simulated the beneficial effects of HM and shifted the gut microbiota of toddlers with CMPA toward that of healthy individuals. In conclusion, this synbiotic-containing pHF might be an appropriate alternative for toddlers with CMPA.

[Retracted] Inhibition airway remodeling and transforming growth factor­ß1/Smad signaling pathway by astragalus extract in asthmatic mice.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that the P­smad2 western blotting data shown in Fig. 7 were strikingly similar to data appearing in different form (namely, the bands appeared in the reverse orientation) in Fig. 4A in another article [Lv Z­D, Na D, Liu F­N, Du Z­M, Sun Z, Li Z, Ma X­Y, Wang Z­N and Xu H­M: Induction of gastric cancer cell adhesion through transforming growth factor­beta1­mediated peritoneal fibrosis. J Exp Clin Cancer Res 29: 139, 2010], which was written by mostly different authors at different research institutes (the author Zheng­Hai Qu did appear as an author on both papers). Owing to the fact that the contentious data in the above article had already been published prior to its submission to International Journal of Molecular Medicine, and due to a lack of overall confidence in the presented data, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they accepted the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Molecular Medicine 29: 564­568, 2012; DOI: 10.3892/ijmm.2011.868].

Relationship between family-related factors and functional constipation among Chinese preschoolers: a case-control study.

BACKGROUND: Constipation is one of the common symptoms in childhood. The prevalence of FC is about 0.5% to 32% and still on the rise according to global statistics. The aim of this study is to explore the associations between family-related factors (e.g., parental conflict, parenting style, and parent-child relationship) and functional constipation of preschool children based on family system theory. METHODS: The study is a case-control survey of preschoolers in China. In total, 108 preschoolers with functional constipation diagnosed with pediatric Rome IV criteria and 324 healthy examination preschoolers without functional constipation were enrolled in the study. Parents completed the following 5 instruments: General information questionnaire, the Parental Conflict Scale, the Parenting Style Questionnaire, the Child-parent Relationship Scale and the Children's Emotional Adjustment Scale-Preschool Version. RESULTS: Nine categories of factors which significantly predicted functional constipation in preschoolers were retained in the final logistic regression model: Second child in birth order (OR = 0.456; 95% CI, 0.229 to 0.910), children picky eating (OR = 2.936; 95% CI, 1.133 to 7.611), bad bowel habits (OR = 2.896; 95% CI, 1.391 to 6.028), parental history of constipation (OR = 3.259; 95% CI, 1.600 to 6.639), parents blaming the child for having a bad bowel movement (OR = 3.788; 95% CI, 1.391 to 10.318), more than 3 h of fathers-child interaction time per day (OR = 0.137; 95% CI, 0.024 to 0.778), parental conflict (OR = 1.981; 95% CI, 0.950 to 3.831), doting or authoritarian parenting style (OR = 1.644; 95% CI, 1.067 to 2.534, OR = 2.481; 95% CI, 1.362 to 4.519), and anxiety control or temper control in children (OR = 0.492; 95% CI, 0.303 to 0.799, OR = 0.189; 95% CI, 0.103 to 0.348). CONCLUSIONS: This study identified the significant associations between family-related factors and functional constipation in preschool children, which provide implications for healthcare professionals to address functional constipation in early childhood using a preventive lens.

Assessment of the effectiveness of parent-targeted interventions for the use of child restraint systems: a systematic review and meta-analysis.

Background: It is essential to implement parent-targeted interventions to increase the use of child restraint systems (CRS) and thus reduce the injuries and deaths of children due to motor vehicle collisions. To optimize future intervention designs, this meta-analysis sought to quantify the effects of parent-targeted interventions and explore potential intervention moderators. Methods: Studies met inclusion criteria if they included a parents-targeted intervention that focused on increasing CRS use for children, published from the inception of the databases to January 2022, were systematically retrieved from the PubMed, Embase, Cochrane library, Web of Science, Sinomed, Wanfang, and CNKI databases. Next, 2 researchers independently screened the retrieved articles, evaluated their quality according to the Cochrane Tool, and extracted the data. Finally, Stata12.0 was used for the meta-analysis. Heterogeneity was examined with I2, stratified analyses, and meta-regression. Results: Of the 1,690 articles retrieved, 9 studies, comprising 22,329 parents of children aged 0-12 years, were ultimately included in the analysis. The results of the meta-analysis showed that the CRS use rate of the intervention group was 1.62 times higher than that of the control group [95% confidence interval (CI): 1.25-2.11, Z=3.616, P<0.001], indicating the positive effect of parent-targeted interventions on promoting the use of CRS. The subgroup analysis found that interventions guided by behavioral theories increased the use of CRS (odds ratio: 1.44, 95% CI: 1.27-1.63, n=5). The difference in the use of CRS between the groups in the studies that were not guided by theories was not statistically significant, indicating that interventions guided by behavioral theories may be the source of the heterogeneity. Risk of bias was low in most studies. Conclusions: It is necessary to conduct interventions with parents to increase the use of CRS. The effects on CRS use appear to differ depending on whether the interventions are guided by behavioral theories. In-depth research needs to be conducted to explore the characteristics of the interventions, especially those guided by different behavioral theories, to reduce child vehicle injuries.

A Review of Three Chinese Cases of Acromicric/Geleophysic Dysplasia with FBN1 Mutations.

OBJECTIVE: This study aims to explore the clinical features and molecular diagnosis of FBN1-related acromelic dysplasia in Chinese patients. METHODS: The clinical and genetic features of three FBN1-related acromicric dysplasia (AD)/geleophysic dysplasia (GD) Chinese patients from two families were reviewed, and comprehensive medical evaluations were performed. Targeted next-generation sequencing was used to detect genetic mutations associated with short statures, including FBN1. Sanger sequencing was used to determine the de novo mutation origin. RESULTS: Patient 1 presented with short stature, short and stubby hands and feet, mild facial dysmorphism, hepatomegaly, delayed bone age and beak-like femoral heads. Patient 2 and this patient's father merely presented with short stature, wide and short hands, and beak-like femoral heads. One novel mutation, c.5272G>T(p.D1758Y), and one known mutation, c.5183C>T(p.A1728V), were identified in these patients. CONCLUSION: The clinical features varied among these patients. The variant c.5272G>T(p.D1758Y) is a novel mutation.

[Analysis of clinical features and genetic variants in an infant with Bloom syndrome].

OBJECTIVE: To analyze the clinical features and genetic variants in a 13-month-old child with Bloom syndrome. METHODS: Clinical data of the child was collected. Genetic variants were detected by high-throughput sequencing and Sanger sequencing. RESULTS: The child was born at full term but was small for gestational age. His clinical features included loss of appetite, severe growth retardation, microcephaly, and small mandible. Genetic testing found that he had carried compound heterozygous c.1068+3A>C and c.1069-1G>C variants of the BLM gene, both of which were unreported previously. CONCLUSION: Bloom syndrome is mainly characterized by severe growth retardation in infancy. The novel variants have expanded the variant spectrum of the BLM gene.

MiR-204-5p Inhibits Transforming Growth Factor-ß1-Induced Proliferation and Extracellular Matrix Production of Airway Smooth Muscle Cells by Regulating Six1 in Asthma.

BACKGROUND: Transforming growth factor-ß1 (TGF-ß1)-in-duced proliferation of airway smooth muscle cells plays critical roles in the development of airway remodeling. Six1 (sine oculis homeobox homolog 1) has been demonstrated to be involved in airway inflammation and remodeling in asthmatic mice. OBJECTIVES: The aim of this work was to investigate the potential role of miR-204-5p in the proliferation and extracellular matrix (ECM) production of airway smooth muscle cells in asthma. METHODS: Real-time PCR was used to measure the expression of miR-204-5p in asthmatic airway smooth muscle cells. Cell viability and apoptosis were detected to evaluate the effect of miR-204-5p on airway smooth muscle cells. Dual-luciferase reporter experiments were applied to identify the target genes of miR-204-5p. RESULTS: MiR-204-5p was downregulated notably in asthmatic airway smooth muscle cells as well as cells stimulated with TGF-ß1. Overexpression of miR-204-5p markedly suppressed the TGF-ß1-induced proliferation of airway smooth muscle cells and the deposition of ECM, whereas the inhibition of miR-204-5p significantly enhanced the proliferation of airway smooth muscle cells and upregulated the level of fibronectin and collagen III. Furthermore, subsequent analyses demonstrated that Six1 was a direct target of miR-204-5p, and Western blot further indicated that miR-204-5p negatively regulated the expression of Six1. Most importantly, the restoration of Six1 expression reversed the inhibitory effect of miR-204-5p on TGF-ß1-induced proliferation and ECM production. CONCLUSIONS: MiR-204-5p inhibits TGF-ß1-in-duced proliferation and ECM production of airway smooth muscle cells by regulating Six1, identifying a potential therapeutic target for preventing airway remodeling in asthma.

Mutations in ASH1L confer susceptibility to Tourette syndrome.

Tourette syndrome (TS) is a childhood-onset neuropsychiatric disorder characterized by repetitive motor movements and vocal tics. The clinical manifestations of TS are complex and often overlap with other neuropsychiatric disorders. TS is highly heritable; however, the underlying genetic basis and molecular and neuronal mechanisms of TS remain largely unknown. We performed whole-exome sequencing of a hundred trios (probands and their parents) with detailed records of their clinical presentations and identified a risk gene, ASH1L, that was both de novo mutated and associated with TS based on a transmission disequilibrium test. As a replication, we performed follow-up targeted sequencing of ASH1L in additional 524 unrelated TS samples and replicated the association (P value = 0.001). The point mutations in ASH1L cause defects in its enzymatic activity. Therefore, we established a transgenic mouse line and performed an array of anatomical, behavioral, and functional assays to investigate ASH1L function. The Ash1l+/- mice manifested tic-like behaviors and compulsive behaviors that could be rescued by the tic-relieving drug haloperidol. We also found that Ash1l disruption leads to hyper-activation and elevated dopamine-releasing events in the dorsal striatum, all of which could explain the neural mechanisms for the behavioral abnormalities in mice. Taken together, our results provide compelling evidence that ASH1L is a TS risk gene.

Scalp acupuncture for children with cerebral palsy: A protocol for a systematic review.

BACKGROUND: Cerebral palsy (CP) describes a group of permanent disorders of movement and posture causing activity limitations, leading the most common movement disorder to children. Scalp acupuncture (SA) is one of several specialized acupuncture techniques, and it has been used widely in China to alleviate several CP symptoms, despite the deficiency of high-quality evidence related to this practice. Therefore, we plan to conduct a protocol of systematic review aimed at systematically reviewing all the clinical evidence on the effectiveness of scalp acupuncture for treating CP in children. METHODS: The following electronic databases will be searched from inception to May 1, 2019 MEDLINE, PubMed, Web of Science, EMBASE, the Cochrane Central Register of Controlled Trials (Cochrane Library), Chinese National Knowledge Infrastructure (CNKI), Chinese Scientific Journals Database (VIP), Wan-fang Database, and Chinese Biomedical Literature Database (CBM). All published English and Chinese articles randomized controlled trials (RTCs) will be included. All types of CP of children in the trials will be included in this study and these individuals will be involved as core searchers to evaluate the efficacy of scalp acupuncture. Rev Man V.5.3 software will be implemented for the assessment of bias risk, data synthesis, subgroup analysis, and meta-analyses if inclusion conditions are met. Continuous outcomes will be presented as mean difference (MD) or standard mean difference (SMD), while dichotomous data will be expressed as a relative risk. RESULTS: The systematic review will synthesize the available knowledge surrounding scalp acupuncture for children with CP. The findings will be synthesized to determine the efficacy and safety of scalp acupuncture for children with CP. CONCLUSION: This protocol will present the evidence of whether scalp acupuncture is an effective intervention for children with CP.

MiR-448-5p inhibits TGF-ß1-induced epithelial-mesenchymal transition and pulmonary fibrosis by targeting Six1 in asthma.

MicroRNAs (miRNAs) are small yet versatile gene tuners that regulate a variety of cellular processes, including cell growth and proliferation. The aim of this study was to explore how miR-448-5p affects airway remodeling and transforming growth factor-ß1 (TGF-ß1)-stimulated epithelial-mesenchymal transition (EMT) by targeting Sine oculis homeobox homolog 1 (Six1) in asthma. Asthmatic mice models with airway remodeling were induced with ovalbumin solution. MiRNA expression was evaluated using quantitative real-time polymerase chain reaction. Transfection studies of bronchial epithelial cells were performed to determine the target genes. A luciferase reporter assay system was applied to identify whether Six1 is a target gene of miR-448-5p. In the current study, we found that miR-448-5p was dramatically decreased in lung tissues of asthmatic mice and TGF-ß1-stimulated bronchial epithelial cells. In addition, the decreased level of miR-448-5p was closely associated with the increased expression of Six1. Overexpression of miR-448-5p decreased Six1 expression and, in turn, suppressed TGF-ß1-mediated EMT and fibrosis. Next, we predicted that Six1 was a potential target gene of miR-448-5p and demonstrated that miR-448-5p could directly target Six1. An SiRNA targeting Six1 was sufficient to suppress TGF-ß1-induced EMT and fibrosis in 16HBE cells. Furthermore, the overexpression of Six1 partially reversed the protective effect of miR-448-5p on TGF-ß1-mediated EMT and fibrosis in bronchial epithelial cells. Taken together, the miR-448-5p/TGF-ß1/Six1 link may play roles in the progression of EMT and pulmonary fibrosis in asthma.

Targeted inhibition of Six1 attenuates allergic airway inflammation and remodeling in asthmatic mice.

Asthma is an inflammatory disease of the airways, characterized by lung eosinophilia, mucus hypersecretion by goblet cells and airway hyperresponsiveness to inhaled allergens. The purpose of this study was to evaluate the effects of Six1 on airway inflammation and remodeling and the underlying mechanisms in a murine model of chronic asthma. Female BALB/c mice were randomly divided into four groups: phosphate-buffered saline control, ovalbumin (OVA)-induced asthma group, OVA+siNC and OVA+siSix1. In this mice model, Six1 expression level was significantly elevated in OVA-induced asthma of mice. Additionally, downregulation of Six1 dramatically decreased OVA-challenged inflammation, infiltration, and mucus production. Moreover, silencing of Six1 resulted in decreased levels of immunoglobulin E and inflammatory mediators and reduced inflammatory cell accumulation, as well as inhibiting the expression of important mediators including matrix metalloproteinase MMP-2 and MMP-9, which is related to airway remodeling. Further analysis indicated that silencing of Six1 can significantly inhibit NF-kB pathway activation in the lungs. .In conclusion, these findings indicated that the downregulation of Six1 effectively inhibited airway inflammation and reversed airway remodeling, which suggest that Six1 represents a promising therapeutic strategy for human allergic asthma.

Transforming growth factor-ß1 induces bronchial epithelial cells to mesenchymal transition by activating the Snail pathway and promotes airway remodeling in asthma.

Airway remodeling is characterized by airway wall thickening, subepithelial fibrosis, increased smooth muscle mass, angiogenesis and an increase in mucous glands, which may lead to a chronic and obstinate asthma with pulmonary function depression. In the present study, we observed substantially thickened lung tissue with extensive fibrosis in ovalbumin-sensitized mice, which was interrelated with transforming growth factor-ß1 (TGF-ß1) expression in bronchoalveolar lavage fluid. In vitro experiments further demonstrated that TGF-ß1 resulted in epithelial-mesenchymal transition (EMT) in bronchial epithelial cells, which was characterized by the expected decrease in E-cadherin expression and the increase in vimentin and α-smooth muscle actin expression, as well as the associated increase in Snail expression at mRNA and protein levels. Furthermore, the downregulation of Snail by small interfering RNA (siRNA) attenuated the TGF-ß1­induced EMT-like phenotype. Of note, a significantly increased synthesis of fibronectin was observed following TGF-ß1 treatment, which further supported the hypothesis that EMT is a pivotal factor in peribronchial fibrosis. In combination, the results indicated that myofibroblasts deriving from bronchial epithelial cells via EMT may contribute to peribronchial fibrosis and that Snail may be an important factor in this phenomenon.

Astragalus extract attenuates allergic airway inflammation and inhibits nuclear factor κB expression in asthmatic mice.

BACKGROUND: Astragalus membranaceus from traditional Chinese herbal medicines previously showed that it possesses a strong anti-inflammatory activity. The purpose of this study was to elucidate the effect of astragalus on allergen-induced airway inflammation and airway hyperresponsiveness and investigate its possible molecular mechanisms. METHODS: Female BALB/c mice sensitized and challenged with ovalbumin (OVA) developed airway inflammation. Bronchoalveolar lavage fluid was assessed for total and differential cell counts and cytokine and chemokine levels. In vivo airway responsiveness to increasing concentrations of methacholine was measured 24 hours after the last OVA challenge using whole-body plethysmography. The expression of inhibitory κB-α and p65 in lung tissues was measured by Western blotting. RESULTS: Astragalus extract attenuated lung inflammation, goblet cell hyperplasia and airway hyperresponsiveness in OVA-induced asthma and decreased eosinophils and lymphocytes in bronchoalveolar lavage fluid. In addition, astragalus extract treatment reduced expression of the key initiators of allergic T(H)2-associated cytokines (interleukin 4, interleukin 5) (P < 0.05). Furthermore, astragalus extract could inhibit nuclear factor κB (NF-κB) expression and suppress NF-κB translocation from the cytoplasm to the nucleus in lung tissue samples. CONCLUSIONS: Taken together, our current study demonstrated a potential therapeutic value of astragalus extract in the treatment of asthma and it may act by inhibiting the expression of the NF-κB pathway.

Inhibition airway remodeling and transforming growth factor-ß1/Smad signaling pathway by astragalus extract in asthmatic mice.

Airway remodeling is characterized by airway wall thickening, subepithelial fibrosis, increased smooth muscle mass, angiogenesis and increased mucous glands, which can lead to a chronic and obstinate asthma with pulmonary function depression. In the present study, we investigated whether the astragalus extract inhibits airway remodeling in a mouse asthma model and observed the effects of astragalus extract on the transforming growth factor-ß1 (TGF-ß1)/Smad signaling pathway in ovalbumin-sensitized mice. Mice were sensitized and challenged by ovalbumin to establish a model of asthma. Treatments included the astragalus extract and budesonide. Lung tissues were obtained for hematoxylin and eosin staining and Periodic acid-Schiff staining after the final ovalbumin challenge. Levels of TGF-ß1 were assessed by immunohistology and ELISA, levels of TGF-ß1 mRNA were measured by RT-PCR, and levels of P-Smad2/3 and T-Smad2/3 were assessed by western blotting. Astragalus extract and budesonide reduced allergen-induced increases in the thickness of bronchial airway and mucous gland hypertrophy, goblet cell hyperplasia and collagen deposition. Levels of lung TGF-ß1, TGF-ß1 mRNA and P-Smad2/3 were significantly reduced in mice treated with astragalus extract and budesonide. Astragalus extract improved asthma airway remodeling by inhibiting the expression of the TGF-ß1/Smad signaling pathway, and may be a potential drug for the treatment of patients with a severe asthma airway.

[Subjective quality of life in children with Tourette syndrome].

OBJECTIVE: This study explored the subjective quality of life in children with Tourette syndrome (TS) in order to provide a basis for more effective interference of TS. METHODS: A total of 174 children with TS (≥ 8 years old) and 186 aged-matched healthy children as controls were enrolled. The subjective quality of life was investigated by a case-control study. RESULTS: The total score of subjective quality of life in the TS group (156.6 ± 21.1) was lower than that in the control group (164.2 ± 21.2; P<0.01). The scores of family life, school life, cognitive component, anxiety experience and depression experience (19.1 ± 3.5 vs 20.7 ± 3.0, 24.1 ± 4.4 vs 26.6 ± 3.2, 90.6 ± 13.3 vs 97.9 ± 15.3, 24.0 ± 4.6 vs 25.1 ± 3.1 and 23.8 ± 4.4 vs 24.7 ± 3.5) in the TS group were lower than those in the control group (P<0.05). The correlation analysis showed that the total score of subjective quality of life in children with TS was negatively related to the age, the course of disease, the severity of symptoms, the total score of child behavior problem and family conflict (r=-0.432, -0.213, -0.869, -0.137, -0.257; P<0.01), while it was positively related to family active-cultural orientation (r=0.084, P<0.01). The multiple step regression analysis indicated that the factors influencing the subjective quality of life in children with TS included the severity of symptoms, age, family conflict and family active-cultural orientation (ß'=-0.787, -0.171, -0.109, 0.106; P<0.01). CONCLUSIONS: The subjective quality of life is not well in children with TS. It is important to control clinical symptoms and improve family environment for the improvement of the subjective quality of life in children with TS.

[Fractal characteristics of mature aerobic granular sludge cultivated by different carbon sources].

In order to characterize the dense and regular level of aerobic granular sludge (AGS) which was cultivated by different carbon sources, the SEM images of mature AGS were used in the study. The calculation process using Photoshop and Fips2 on fractal characteristics of these granules was built. The lowest box-counting dimension was bulking aerobic granular sludge cultivated by glucose (No. 1), which was 1.794 +/- 0.011; the box-counting dimension of AGS cultivated by peptone (No. 3) and domestic sewage (No. 5) were reached up to 1.866 +/- 0.018 and 1.880 +/- 0.015. The boundary box-counting dimension of the AGS was also calculated and the average value was 1.14. The AGS cultivated by beer (No. 6) was more regular in shape and the boundary box-counting dimension was 1.115 +/- 0.003. The AGS cultivated by landfill leachate (No. 7) was the most irregular in shape. This study indicates that fractal dimension provides an approach for quantification of dense and regular level of AGS, furthermore, it could be used to characterize the status of AGS, such as bulking.

[Formation and reaction mechanism of simultaneous nitrogen and phosphorus removal by aerobic granular sludge].

Aerobic granular sludge (AGS) for simultaneous nitrogen and phosphorus removal (SNPR) was cultivated and studied in two lab-scale sequencing batch reactors (named as A & B) treating real domestic wastewater, additional carbon (sodium propionate and sodium acetate for A, glucose for B) was added to make the ratio of COD:N:P as 360:60:6, good SNPR was achieved at normal (18-27 degrees C) and low temperature (9-13 degrees C). The microbial community composition and distribution, distribution of cells and extracellular polymeric substances (EPS) and morphologies of the AGS were investigated using fluorescence in situ hybridization (FISH), in situ fluorescent staining and scanning electron microscope (SEM), respectively. FISH results showed that ammonium-oxidizing bacteria comprised 12% of all the bacteria and were mainly located at the outer parts of the granules; phosphates accumulating organisms comprised 40% of all the bacteria and were mainly located in the inner parts of the granules. Nitrification was the rate controlling step; denitrifying phosphate accumulating organisms inside the granular sludge might be responsible for denitrification in the aerobic phase, which enabled effective SNPR. Live/dead fluorescent staining results showed that dead cells were distributed throughout the granules and live cells were principally distribution of polysaccharide (including alpha-mannopyranosyl, alpha-glucopyranosyl sugars and beta-D-glucopyranose polysaccharides) of EPS in AGS were influenced by different carbon sources, but the contents and distributions of protein and lipids were not, the contents of protein was the largest. Polysaccharide was responsible for the formation and maintenance of aerobic granular sludge. SEM results showed that bacilli and cocci were the main bacteria in the granules of A and B, respectively. Carbon sources affected the bacteria type and the SNPR efficiency, sodium propionate and sodium acetate were better than glucose.

[Simultaneous nitrogen and phosphorus removal by aerobic granular sludge at normal and low temperatures].

The simultaneous nitrogen and phosphorus removal (SNPR) from real domestic wastewater using aerobic granular sludge (AGS) has been investigated. The effect of temperature changes on the SNPR of AGS was also studied. Two lab-scale sequencing batch reactors (SBRs), named as A and B, were operated for 254 days in alternating anaerobic-aerobic mode. The AGS was cultivated in 20 days using settling time as selection pressure. From the 42nd day onwards, additional carbon (acetate and peptone for A, glucose for B) were added to make the ratio of COD:N:P as 360:60:6. The SNPR by AGS was achieved for 4 months, and the removal ratios of ammonia, total inorganic nitrogen and phosphorus of A were reached to 98.42%, 74.25% and 94.79%, respectively; the removal ratios of ammonia, total inorganic nitrogen and phosphorus of B were reached to 99.45%, 75.96% and 95.60%, respectively. Then, the temperature decreased to 9-13 degrees C, which seriously affected the efficiency of SNPR. The removal efficiencies of SNPR of reactor A and B were recovered after 35 days and 49 days, respectively, and the removal ratios of ammonia, total inorganic nitrogen and phosphorus of A were reached to 96.33%, 79.49% and 99.68%, respectively; the removal ratios of ammonia, total inorganic nitrogen and phosphorus of B were reached to 93.85%, 76.44% and 98.44%, respectively. PAOs and DNPAOs together completed the phosphorus removal and made phosphorus removal faster than nitrogen removal. Nitrification was the rate limiting step for SNPR. The low temperature made the anoxic zone in AGS decrease, together with the absence of PHAs, made denitrification could not complete.