CYP4G subfamily genes mediate larval integument development in Spodoptera frugiperda.

Cytochrome P450 (CYP) 4G subfamily is closely related to the synthesis of cuticular hydrocarbons, leading to the enhanced desiccation and insecticide resistance of pests. However, functions of CYP4Gs in larval integument development remain unknown in Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), which is a major transboundary migratory pest and become a common pest in China. On the basis of the genome and transcriptome datasets of S. frugiperda, CYP4G74, CYP4G75, CYP4G108, and CYP4G109 were identified, which contained the conserved domains of P450s and CYP4Gs. The spatial and temporal expression analysis showed that CYP4G74 and CYP4G75 were significantly highly expressed in adults and larval integuments, while CYP4G108 and CYP4G109 had low expressions in larval integuments. After silencing CYP4G74 and CYP4G75 by RNA interference, abnormal integument development occurred in larvae, some of which became smaller and dead, indicating important roles of CYP4G74 and CYP4G75 in the synthesis and development of integuments. The results clarify the functions of CYP4Gs in S. frugiperda and provide potential targets for the control of this pest.

Design of ultra-wide-field scanning laser fundus imaging system with cascaded conicoid mirrors.

We propose and design a multi-stage cascaded scanning laser ophthalmoscope (SLO) for ultra-wide field (UWF), which uses conicoid mirrors, constructed by conjugation of pupil plane. The vergence uniformity and the angular magnification of a cascaded conicoid mirrors (CCM) system are analyzed recursively and optimized preliminarily to achieve high quality imaging with UWF, and the optimal system with the model eye are obtained by simulation and optimization. Two-stage and three-stage cascaded systems are designed with this method, and the formulas of beam vergence and angular magnification are obtained by theoretical derivation. As compared to the two-stage CCM system, the proposed three-stage cascaded UWF SLO has superior performance in imaging quality. Its average RMS radius of spot diagram is calculated to be 26.372 µm, close to the diffractive limit resolution. The image resolution of human retina can be up to 30 µm with 135° FOV in theory. The three-stage cascaded SLO is more suitable for UWF fundus imaging. This study will be helpful for early screening and accurate diagnosis of various diseases in the peripheral retina.

Aerosol exchange between pressure-equilibrium rooms induced by door motion and human movement.

It is now widely recognised that aerosol transport is major vector for transmission of diseases such as COVID-19, and quantification of aerosol transport in the built environment is critical to risk analysis and management. Understanding the effects of door motion and human movement on the dispersion of virus-laden aerosols under pressure-equilibrium conditions is of great significance to the evaluation of infection risks and development of mitigation strategies. This study uses novel numerical simulation techniques to quantify the impact of these motions upon aerosol transport and provides valuable insights into the wake dynamics of swinging doors and human movement. The results show that the wake flow of an opening swinging door delays aerosol escape, while that of a person walking out entrains aerosol out of the room. Aerosol escape caused by door motion mainly happens during the closing sequence which pushes the aerosols out. Parametric studies show that while an increased door swinging speed or human movement speed can enhance air exchange across the doorway, the cumulative aerosol exchange across the doorway is not clearly affected by the speeds.

Expressions and functions of RR-1 cuticular protein genes in the integument of Mythimna separata.

As the most outer layer between itself and the environment, integuments are necessary for insects with various important functions. Cuticular proteins (CPs) are the main components in integuments, while the functions of CP genes remain unknown in Mythimna separata (Walker), which is a devastating agricultural pest. In this study, 79 CP genes were identified from the transcriptomes of larval integuments, 57 of which were from the family containing conserved Rebers & Riddiford (R&R) consensus (CPR family). Amongst these CPRs, 44 genes belonged to the subfamily with RR-1 motif (RR-1 genes) and clustered into three clades, with the top 15 most abundant RR-1 genes identified based on fragments per kilobase per million mapped fragments (FPKM) values. RT-qPCR analysis showed that most of RR-1 genes such as MsCPR1-4 were highly expressed at larval stages and in their integuments. The expression levels of RR-1 genes were generally decreased at the beginning but increased at the late stage of molting process. RNAi was applied for six RR-1 genes, and MsCPR1-4 were knocked down significantly. Silence of MsCPR2 resulted in abnormal integument formed after molting, while knockdown of MsCPR3 and MsCPR4 led to failure of molting, respectively. No phenotype was obtained for the RNAi of MsCPR1. Therefore, the expression of RR-1 genes and their functions were analyzed in the development of integuments in M. separata, providing new insights of RR-1 genes and potential targets for the development of growth regulators and new insecticides for M. separata.

One-year myopia control efficacy of cylindrical annular refractive element spectacle lenses.

PURPOSE: To evaluate the 1-year myopia control efficacy of a spectacle lens with annular cylindrical microstructures. METHODS: A total of 118 consecutive eligible children aged 8-12 years with -1.00 D to -4.00 D of spherical component myopia and <1.50 D astigmatism were enrolled between August 2020 and November 2020 at the Eye Hospital of Wenzhou Medical University. Participants were randomly assigned to wear cylindrical annular refractive element (CARE) (n = 61) or single-vision (n = 57) spectacle lenses. Cycloplegic autorefraction (spherical equivalent refraction [SER]) and axial length (AL) were measured at baseline and 6-month intervals. Adaptation and compliance questionnaires were administered during all visits. RESULTS: Among 118 randomized participants, 96 (81.4%) were included in the analyses (mean [SE] age, 10.4 [0.6] years; 49 [51.0%] were female; mean [SE] spherical equivalent refractive error, -2.67 [0.66] D; mean [SE] axial length, 24.75 [0.77] mm). Adjusted 1-year myopia progression was -0.56 D for CARE and -0.71 D for single-vision spectacle lenses. The difference in progression was 0.14 D (95% CI, -0.04 to 0.32) for CARE vs single vision. Adjusted 1-year eye growth was 0.27 mm for CARE and 0.35 mm for single vision. The difference in eye growth was 0.09 mm (95% CI, -0.15 to -0.02) for CARE vs single vision. All groups adapted to their lenses with no reported adverse events, complaints, or discomfort. CONCLUSIONS: Among children with myopia, treatment with cylindrical annular refractive element spectacle lenses significantly reduced the rate of axial elongation over 1 year compared with single-vision spectacle lenses.

Surface astigmatism correction using segmented freeform surfaces for a progressive addition lens.

The undesired distribution of irregular surface astigmatism (SA) on the freeform surface has been the major concern of progressive addition lens (PAL) design. Herein, we proposed a segmented freeform surface (SFS) construction method, which relies on the lines of curvature to rule the surface segmentation and then eliminates the difference between principal curvatures to correct the SA. Based on ray tracing and numerical simulation results, the SFS-PAL design has superior performance in image quality within a dynamic field of view over the conventional freeform PAL. To verify the feasibility and the real performance of the new design, we used the diamond turning method with a fast tool servo to realize the rapid prototyping, and then used injection molding for the mass production of the high-quality SFS-PALs.

Regional Changes in Posterior Corneal Surface During a 6-Month Follow-up Period After tPRK, FS-LASIK, and SMILE.

PURPOSE: To investigate changes in corneal curvature in different zones of the posterior corneal surface during a 6-month follow-up period after transepithelial photorefractive keratectomy (tPRK), femtosecond laser-assisted excimer laser in situ keratomileusis (FS-LASIK), and small incision lenticule extraction (SMILE). METHODS: The study included a total of 202 eyes, including 65, 77, and 60 that underwent tPRK, FS-LASIK, and SMILE, respectively. Elevation data for the posterior surface were obtained preoperatively (pre), as well as 1 week (pos1w), 1 month (pos1m), 3 months (pos3m), and 6 months (pos6m) postoperatively. Changes in posterior corneal curvature (M) were analyzed in the central (diameter: 0 to 3 mm), paracentral (diameter: 3 to 6 mm), and peripheral (diameter: 6 to 9 mm) regions. RESULTS: Over all follow-up periods, the central region of the posterior surface in all patients became flatter (P < .05), with FS-LASIK showing the largest change, whereas the paracentral and peripheral regions became steeper. The posterior curvature changes between pre and pos6m, determined before and after correction for ablated stromal depth, tended to follow similar trends in the three regions and after the three surgeries. There was also no significant correlation (P > .05) between the changes in the mean curvature (M, recorded between pre and pos6m) and each of the refractive error corrections, the changes in spherical aberration postoperatively, the optical zone diameter, ablated stromal depth, and residual stromal bed thickness in the central and peripheral regions, but the correlation was significant in the paracentral region. CONCLUSIONS: The postoperative changes in posterior corneal shape followed different trends in the central, paracentral, and peripheral regions. The FS-LASIK group exhibited the most notable changes in posterior corneal curvature, especially in the central region. These changes were statistically correlated with variations in spherical aberration, and ablated and residential stromal thickness in the paracentral region. [J Refract Surg. 2022;38(11):708-715.].

Circulating soluble CD36 as a novel biomarker for progression and prognosis of HBV-related liver diseases.

Background: Our previous study suggested CD36 may be a positive regulator of hepatitis B virus (HBV) replication in vitro. Therefore, the present study aimed to investigate whether circulating soluble CD36 (sCD36) could serve as a diagnostic and prognostic biomarker for HBV-related liver diseases based on the clinic collected data. Methods: A total of 282 subjects were divided into healthy controls (HC, n = 47), chronic hepatitis B (CHB, n = 68), HBV-related liver cirrhosis (HBV-LC, n = 167). Soluble CD36 in plasma was measured by ELISA, and monocyte or platelet CD36 expression was determined by flow cytometry. Results: There was a step-wise increase of sCD36 with the progression of chronic HBV infection, and it was the highest in the HBV- LC group with liver failure (1.50, IQR:1.04-2.00) as compared with HC (0.38, IQR:0.27-0.38), CHB (0.75, IQR:0.40-1.13), and HBV-LC without liver failure (1.02, IQR,0.61-1.35) group. Circulating sCD36 was not correlated with serum HBV DNA levels, but correlated with liver function parameters. Additionally, ROC analysis confirmed sCD36 could be used to predict liver failure for HBV-LC patients, which yielded an AUC of 0.775 with 71.0% sensitivity and 72.2% specificity. Multivariate logistic regression analysis revealed sCD36 is an independent risk factor in predicting liver failure. Moreover, plasma sCD36 in HBV-LC patients was significantly correlated with prognostic indices, including MELD, MELD-Na and CHILD-PUGH scores. On the other hand, CD36 expression on monocytes or platelets was positively correlated with plasma sCD36 levels, whereas they were not strongly associated with the disease severity. Conclusion: Circulating sCD36 could be used as a novel noninvasive biomarker for predicting liver failure and prognosis in chronic HBV infected patients.

HIF-2α-induced upregulation of CD36 promotes the development of ccRCC.

Clear cell renal cell carcinoma (ccRCC) is characterized by the abundance of lipid droplets and the activation of the hypoxia-inducible factor (HIF) signaling pathway. However, the lipid reprogramming induced by HIF signaling in ccRCC is not fully understood. In this study, we found that the fatty acid receptor CD36 was highly expressed in human ccRCC tissues and ccRCC cell lines. CD36 overexpression increased fatty acid uptake and lipid droplet formation, and enhanced the proliferation and migration of ccRCC cells in a DGAT1-dependent manner. In contrast, the disruption of endogenous CD36 showed the opposite effects. The upregulated expression of CD36 in ccRCC was associated with hypoxia and HIF-2α activation. Furthermore, we identified CD36 as a new target of the transcription factor HIF-2α. The knockdown of CD36 in ccRCC cells reduced lipid accumulation and also blocked the tumor-promoting effects induced by HIF-2α under hypoxia. Our findings suggest that hypoxia-dependent HIF-2α promotes the remodeling of lipid metabolism and the malignant phenotype of ccRCC via CD36, providing a certain theoretical basis for clarifying the mechanism of ccRCC.

CD36-mediated metabolic crosstalk between tumor cells and macrophages affects liver metastasis.

Liver metastasis is highly aggressive and treatment-refractory, partly due to macrophage-mediated immune suppression. Understanding the mechanisms leading to functional reprogramming of macrophages in the tumor microenvironment (TME) will benefit cancer immunotherapy. Herein, we find that the scavenger receptor CD36 is upregulated in metastasis-associated macrophages (MAMs) and deletion of CD36 in MAMs attenuates liver metastasis in mice. MAMs contain more lipid droplets and have the unique capability in engulfing tumor cell-derived long-chain fatty acids, which are carried by extracellular vesicles. The lipid-enriched vesicles are preferentially partitioned into macrophages via CD36, that fuel macrophages and trigger their tumor-promoting activities. In patients with liver metastases, high expression of CD36 correlates with protumoral M2-type MAMs infiltration, creating a highly immunosuppressive TME. Collectively, our findings uncover a mechanism by which tumor cells metabolically interact with macrophages in TME, and suggest a therapeutic potential of targeting CD36 as immunotherapy for liver metastasis.

Effects of black carbon aerosol on air quality and vertical meteorological factors in early summer in Beijing.

Black carbon (BC) aerosols have effects on the atmospheric thermal vertical structure due to its radiation absorption characteristics, hereby influencing the boundary layer characteristics and pollutant diffusion. This study focuses on the BC effects under different atmospheric conditions on air quality and vertical meteorological conditions. Four days flight observation combined with surface wind profiler radar data were used to investigate the vertical profiles of BC and wind speed over Beijing urban area in early summer. The vertical profiles of BC concentration and wind speed in the boundary layer had a negative correlation, both having abrupt changes near the boundary layer height under stagnant weather conditions. The chemical transport model showed the increase of BC under stagnant conditions could cause aggravation of the stability of the boundary layer, thereby increasing the accumulation of pollutants. In particular, BC leads to the changes in the temperature profile, which will modify relative humidity and indirectly lead to the changes in the vertical profile of aerosol optical properties. However, if the early accumulation of BC was absent under more turbulent conditions, the effects of BC on air quality and meteorological conditions were limited.

Deformed Mediated Larval Incisor Lobe Development Causes Differing Feeding Behavior between Oriental Armyworm and Fall Armyworm.

Mandibular incisor lobes are important for insect feeding behavior, living habits and niche. However, the molecular regulation of insect incisor lobe development remains unknown. In this study, we found that two maize pests, oriental armyworm Mythimna separata and fall armyworm Spodoptera frugiperda, have different feeding patterns in maize, which are closely associated with the different development patterns of their incisor lobes. Different from first to sixth instar S. frugiperda, which feed on leaf tissues and whorls with sharp incisor lobes, older instars of M. separata feed from leaf margins with no incisor lobes. Hox gene Deformed (Dfd) is important for head appendages, but its function in incisor lobe development is not clear. Here, Dfds were identified from two armyworm species, and both were expressed highly in heads and eggs. Interestingly, the expression levels of MsDfd were relatively high in larval mandibles and decreased dramatically from fourth-instar mandibles in M. separata. Knockdown of MsDfd resulted in malformed mandibles with no incisor lobe in M. separata, making the larvae unable to perform window-feeding. However, RNAi of SfDfd did not affect the mandibles and window-feeding pattern of S. frugiperda, indicating the different roles of Dfd in these two species. Moreover, the mortality of new first instar M. separata increased after feeding dsMsDfd but did not for S. frugiperda feeding dsSfDfd. These findings revealed that Dfd mediated the larval mandibular incisor lobe morphology, affecting its feeding pattern in M. separata, broadening the knowledge of Dfd functions in insect mandibles and feeding behavior.

Normative Data and Determinants of Macular, Disc, and Peripapillary Vascular Density in Healthy Myopic Children Using Optical Coherence Tomography Angiography.

Objective: To establish a normative database for the vascular density (VD) in macular, disc, and peripapillary regions in healthy myopic children and to evaluate associated ocular features with optical coherence tomography angiography (OCTA). Methods: This was an observational, prospective and cross-sectional study. 776 Chinese healthy myopic children (375 boys and 401 girls) were enrolled, mean (±SD) age 9.84 ± 1.98 (range 6-16) years. En-face angiogram OCTA was performed on 6 mm × 6 mm retinal and 4.5 mm × 4.5 mm disc regions. VD measurements in the macular retina were segmented into the four regions: superficial capillary plexus (SCP), intermediate capillary plexus (ICP), deep capillary plexus (DCP), and choriocapillaris (CC). Correlations between macular, disc, and peripapillary VD and possible influencing factors [age, gender, axial length (AL), spherical equivalent refraction (SER), right/left eye, and signal strength index (SSI)] were assessed by Pearson's correlation and multivariate regression analysis. Results: For macular scans, the corrected VD in the ICP region was (48.25 ± 4.24)% for the whole macular retina. The macular ICP VD in most sections was lower than the SCP, DCP, and CC (all P < 0.001). The corrected VD in CC was (72.96 ± 4.42)% for the whole macular retina. The macular CC VD in every section was all higher than the SCP, ICP, and DCP (all P < 0.001). The size of foveal avascular zone (FAZ) and foveal VD 300 (FD-300) was 0.28 mm2± 0.10 mm2 and (58.43 ± 4.17)% respectively. For disc scans, the corrected VD was (58.04 ± 2.73)% for the whole disc area. Both AL and SER were strongly correlated with ICP, DCP, and CC VD in all regions (all P < 0.01). Larger SSI was correlated with a lower VD in the SCP and ICP, and a higher VD in DCP and CC (all P < 0.01). Conclusion: Vascular density values provide large scale normative data on macular, disc, and peripapillary vascular parameters in a large sample of healthy myopic children with OCTA measured in the four different retinal plexuses and regions. The VD in different regions had various influencing factors; mainly a close correlation with AL and SSI.

CD36 promotes de novo lipogenesis in hepatocytes through INSIG2-dependent SREBP1 processing.

OBJECTIVE: Enhanced de novo lipogenesis (DNL) in hepatocytes is a major contributor to nonalcoholic fatty liver disease (NAFLD). Fatty acid translocase (FAT/CD36) is involved in the pathogenesis of NAFLD through facilitating free fatty acids uptake. Here, we explored the effects of CD36 on DNL and elucidated the underlying mechanisms. METHODS: We generated hepatocyte-specific CD36 knockout (CD36LKO) mice to study in vivo effects of CD36 on DNL under high-fat diet (HFD). Lipid deposition and DNL were analyzed in primary hepatocytes isolated from CD36LKO mice or HepG2 cells with CD36 overexpression. RNA sequence, co-immunoprecipitation, and proximity ligation assay were carried out to determine its role in regulating DNL. RESULTS: Hepatic CD36 expression was upregulated in NAFLD mice and patients, and CD36LKO mice exhibited attenuated HFD-induced hepatic steatosis and insulin resistance. We identified hepatocyte CD36 as a key regulator for DNL in the liver. Sterol regulatory element-binding protein 1 (SREBP1) and its downstream lipogenic enzymes such as FASN, ACCα, and ACLY were significantly downregulated in the liver of HFD-fed CD36LKO mice, whereas overexpression CD36 stimulated insulin-mediated DNL and lipid droplet formation in vitro. Mechanistically, CD36 was activated by insulin and formed a complex with insulin-induced gene-2 (INSIG2) that disrupts the interaction between SREBP cleavage-activating protein (SCAP) and INSIG2, thereby leading to the translocation of SREBP1 from ER to Golgi for processing. Furthermore, treatment with 25-hydroxycholesterol or betulin molecules shown to enhance SCAP-INSIG interaction, reversed the effects of CD36 on SREBP1 cleavage. CONCLUSIONS: Our findings identify a previously unsuspected role of CD36 in the regulation of hepatic lipogenic program through mediating SREBP1 processing by INSIG2, providing additional evidence for targeting CD36 in NAFLD.

Underlying function regulators of anaerobic granular sludge: Starvation and dormancy.

Anaerobic granular sludge (AnGS) is the core of anaerobic granular sludge bed system. In this study, the effect and its mechanism of stopping substrate supply on function of AnGS were investigated. The cutoff of exogenous substrate supply triggered AnGS to enter the dormant state. Some methanization microorganisms sporulated. The number and activity of methanization microorganisms based on 16S rDNA and 16S rRNA/16S rDNA ratio declined and stayed at 45.5% and 0.06% (bacteria), 48.7% and 0.39% (archaea) of the initial vegetative value, respectively. The resuming of exogenous substrate supply promoted AnGS to restore the vegetative state. The spores disappeared. The specific methanization activity of AnGS returned to the original level of 35.82 mL-CH4/g-VSS·d, but the delay time for gas production (DTGS) was prolonged from 9.54 to 18.04 h (0-132 d). The dormancy of methanization microorganisms was the main cause for the fluctuation of apparent function and the stability of intrinsic function of AnGS under starvation stress. The dormancy stabilized the structure and sustained the methanization community of AnGS via the reduction of EPS (structure binder/energy reserve) consumption.

Effect of temperature decrease on anammox granular sludge: Shock and adaptation.

Cryopreservation is one of the effective methods for the preservation of anammox granular sludge (AnGS). However, the effects of cooling pretreatment on AnGS are still unclear. In this study, the effects of temperature decrease on AnGS property were investigated by designing different cooling modes: constant at room temperature 20-25 °C (CK), sharp cooling to 4 °C (S4), -20 °C (S20) and stepwise cooling to 4 °C (A4), -20 °C (A20). The results showed that compared with CK, the cooling modes in S4, S20, A4 and A20 improved the physical preservability of AnGS, slowing down the changes of color, shape and structure; and elevated the preservation rate of functional bacteria Planctomycetes (phylum level) and Candidatus Brocadia (genus level). The preservation rate of live cells in different experimental groups was 48.4 ± 1.8%(CK), 61.1 ± 3.3%(S4), 37.8 ± 0.8%(S20), 81.7 ± 4.8%(A4), 61.9 ± 3.1%(A20), respectively. The Anaerobic Ammonium Oxidation Bacteria (AnAOB) in the stepwise cooling mode (A4 and A20) were found to enter the dormant state and form "dormant zoogloea", while the AnAOB in the sharp cooling mode (S4 and S20) were observed to enter the shock state with a little change. The findings in this work (especially the dormant state of AnAOB) are helpful to understand the effect of temperature decrease on AnGS and to promote the development of AnGS preservation technology.

Potential of anammox process towards high-efficient nitrogen removal in wastewater treatment: Theoretical analysis and practical case with a SBR.

Anammox process has been widely applied in the wastewater nitrogen removal for its high rate and low cost. However, few researches reported the process potential in treating low-strength nitrogen wastewater. In this study, a sequencing batch reactor (SBR) was taken to explore the feasibility of low-strength nitrogen wastewater treatment by anammox process in theory and practice. The practical operation indicated that the effluent with satisfactory quality (1.90 ± 0.70 mg-TN·L-1) could be achieved, when the SBR was fed with low-strength nitrogen influent (6.20 ± 0.45 mg-NH4+-N·L-1 and 7.96 ± 0.59 mg-NO2--N·L-1). The hydraulic retention time (HRT), nitrogen removal efficiency, nitrogen removal rate (NRR) and hydraulic loading rate of SBR were 5.42 h, 86.5%, 0.054 kg-N·m-3·d-1 and 4.43 m3·m-3·d-1 during the 79-day operation, respectively. The theoretical analysis revealed the potential of anammox SBR. When SBR is stably operated, the maximum NRR would be 0.062 kg-N·m-3·d-1 if the effluent nitrogen was required to be as low as 3 mg·L-1. The NRR value is feasible for engineering. However, considering the lower specific substrates utilization rate in practice, the maximum stable NRR was calibrated and found inefficient afterwards. In order to improve the potential of anammox process, the reactors without back mixing and with periodic bioaugmentation should be taken in priority for the engineering applications. In particular, the bioaugmentation frequency and single addition amount were calculated as 7 d and 0.3 g-VSS·L-1, respectively. The results may provide guidance for the development of high-efficient and stable nitrogen removal process under low-strength condition.

Deciphering correlation between permeability and size of anammox granule: "pores as medium".

Anammox granular sludge bed technology has been widely applied for its attractive advantages. Efficient mass transfer is an important factor for the anammox granules to play their role. In this study, steady-state anammox granules were used to investigate the correlation between the permeability and granule size with the granule pore as pivot. The results of size distribution showed that the anammox granules could be divided into 6 groups: 200-500 µm (I), 500-1000 µm (II), 1000-1500 µm (III), 1500-2000 µm (IV), 2000-3000 µm (V) and ≥3000 µm (VI). The results of settling experiment demonstrated that the permeability of anammox granules was negatively correlated with the granule size. The fluid collection efficiency declined from 39.4% to 9.3% for granule group I to III, and further to 0 for granule group IV to VI (granule size was larger than 1.5 mm). The observation of micro-CT revealed that the pore structure of anammox granules varied significantly with the increase of granule size, forming a denser surface layer and sparser interior. The chemical analysis and microscopic observation indicated that the pore plugging of surface layer by cell proliferation and EPS secretion was the main cause for the permeability deterioration. The findings of this study will help to understand the mass transfer of anammox granules and promote the development of anammox processes.

Performance of DOuble Circulation Anaerobic Sludge bed reactor: Biomass self-balance.

The calcification of Anaerobic Granular Sludge is a serious problem in the application of anaerobic methanization biotechnology. Regular replacement of calcified sludge with exogenous sludge is an effective method, but it is costly and troublesome. A new DOuble Circulation Anaerobic Sludge bed reactor was developed for the enhanced production of endogenous sludge to self-balance the discharge of calcified sludge. The sludge washout rate was demonstrated to fall by 45% and the sludge proliferation rate was shown to rise by 230%, offsetting the regular discharge of calcified sludge. The zoogloea in 100 µm dimension was revealed to be the intermediate component of sludge. The sludge proliferation mode was proposed as follows: (i) Growth of sludge; (ii) Self-cracking of sludge to release fragmental sludge; (iii) Migration of fragmental sludge by self-floatation; (iv) Accumulation of suspended sludge in the sedimentation chamber; (v) Re-granulation of suspended sludge with the aid of Venturi effect.