Influence of intergenerational relationships on depressive symptoms in ageing Chinese adults in Hong Kong: Mediating effects of sense of loneliness.

BACKGROUND: Mental health of older adults could be positively predicted by harmonious parent-adult children relationships, although the mechanism has not been sufficiently demonstrated. This study employed sense of loneliness as mediator to examine the influence of multiple domains of intergenerational relationships on depressive symptoms in ageing Chinese adults. METHODS: Data was extracted from a representative survey in Hong Kong among Chinese adults aged over 50 with at least one adult child (n = 801). Four key domains (structural-associational, consensual-normative, affectual closeness, and intergenerational conflict) were adopted to measure the intergenerational relationship quality. Depressive symptoms were assessed using the five-item Geriatric Depression Scale. The mediating role of sense of loneliness in the association between intergenerational relationships and depressive symptoms were tested by the PROCESS macro in SPSS. RESULTS: The influence of overall intergenerational relationship quality and its four subdomains on depressive symptoms were significantly mediated by sense of loneliness. Among the four domains, affectual closeness presented the strongest association with older people's depressive symptoms. The effect of intergenerational conflict on depressive symptoms was completely mediated by sense of loneliness, and the effects of remaining three domains were partially mediated. The domain of consensual-normative solidarity received the lowest rating by Chinese older adults. CONCLUSION: The psychological pathway that loneliness links intergenerational relationships and depressive symptoms was supported in this study. With respect to improving intergenerational relationships, enhancing affectual connection between older parents and adult children was essential to prevent mental problems. This study calls for more attention to the protective role of diverse social relationships in improving mental health through multiple pathways.

Hepatitis C virus core protein increases Snail expression and induces epithelial-mesenchymal transition through the signal transducer and activator of transcription 3 pathway in hepatoma cells.

AIM: Aberrant expression of Snail, a mediator of epithelial-mesenchymal transition (EMT), is crucial for cancer invasiveness and metastasis. Although hepatitis C virus (HCV) core protein has been implicated in hepatocarcinogenesis, the relationship between HCV core and Snail expression has not been clarified. METHODS: HepG2 and Huh7 stable cell lines were established by transfection with pcDNA-HCVc. HepG2-HCVc and Huh7-HCVc cells were co-administered with AG490. Cell migration and invasiveness were tested. STAT3 and Snail expression was analyzed by Real-time PCR and Western blot. RESULTS: We found that HCV core is capable of increasing Snail expression and inducing EMT in hepatoma cells. HCV core-induced Snail expression was accompanied by activation of signal transducer and activator of transcription 3 (STAT3), inhibition of STAT3 abrogated HCV core-induced Snail expression and EMT. Furthermore, chromatin immunoprecipitation showed that phosphorylated STAT3 directly binds to the Snail promoter. CONCLUSION: Collectively, these results suggest that HCV core would play a role in hepatocellular carcinoma invasiveness and metastasis by activating the STAT3 pathway, increasing Snail expression and subsequently triggering EMT. These findings would advance the understanding of HCV-mediated invasiveness and metastasis, and might provide a new potential therapeutic target for HCV-related hepatocellular carcinoma.

Overexpression of miR-122 promotes the hepatic differentiation and maturation of mouse ESCs through a miR-122/FoxA1/HNF4a-positive feedback loop.

BACKGROUND & AIMS: microRNA-122 is the only identified liver-specific miRNA and plays a crucial role in liver development, maintenance of hepatic homeostasis as well as tumourigenesis. In our previous differentiation of ESCs into hepatocytes, microRNA-122 (miR-122) was expressed at a relatively low level. Here, we aim to elucidate the effect and underlying mechanisms of miR-122 during differentiation of ESCs into hepatocytes. METHODS: Mouse ESCs were initially induced towards HPCs by activin A, FGF-4 and sodium butyrate and were subsequently transfected with a recombinant adenovirus expressing vector pAV.Ex1d-CMV>miR-122/IRES/eGFP 9 days after induction. Cells were analysed by real-time PCR, immunofluorescence, flow cytometry, microscopy and functional assays. Furthermore, microarray analysis was performed. RESULTS: We demonstrated that overexpression of miR-122 could effectively promote hepatic differentiation and maturation, as assessed by morphological and functional tests. The microarray analysis revealed that 323 genes were down-regulated, whereas 59 were up-regulated. Particularly, two liver-specific transcription factors, FoxA1 and HNF4a, were significantly up-regulated. Moreover, the expression of E-cadherin was dramatically increased and the proliferation of HPCs was suppressed, whereas knockdown of FoxA1 reduced E-cadherin expression and increased the proliferation of HPCs. In addition, the expression levels of FoxA1, HNF4a and E-cadherin in time-course transfection experiments with miR-122 were not significantly increased except in cells in which transfection with miR-122 occurred 9 days after induction. CONCLUSION: Overexpression of miR-122 at an appropriate stage could promote hepatic differentiation and maturation by regulating the balance between proliferation and differentiation, as well as the balance between EMT and MET, partially through a miR-122/FoxA1/HNF4a-positive feedback loop.

[Simulating Changes in Ecosystem Carbon Storage and Analyzing Influencing Factors in the Central Zhejiang under the Background of LUCC].

Central Zhejiang is the key ecological function area of Zhejiang Province and an important part of the Qiantang River ecological corridor. Conducting simulations of ecosystem carbon storage changes and analyzing influencing factors under land-use changes in this region is of significant importance for achieving the goals of "peak carbon" and "carbon neutrality" at an early stage. This study, based on the land use data of five periods from 1980 to 2020 in central Zhejiang, coupled the GeoSOS-FLUS and the InVEST model to analyze the spatiotemporal changes in carbon storage in the region over time and project for the year 2030. The study also explored the impact of socio-economic and natural factors on changes in carbon storage. The results showed that： ① Between 1980 and 2020, urban construction land in the Zhejiang Central Region increased by 289.91%, with cultivated land and forest land being the main sources, leading to a 3% decrease in ecosystem carbon storage, amounting to 588.88×104 tons. ② High-value areas of carbon storage in the Zhejiang Central Region were concentrated in P'an-an County, Jinyun County, Wuyi County, and other areas. ③ Under the two different scenarios of natural development and ecological protection, carbon storage in the research area was projected to decrease by 1.05% and 0.05%, respectively, by 2030 compared to that in 2020. ④ Natural factors dominated the distribution of carbon storage, but their influence was gradually decreasing over time. The impact of socio-economic factors was increasing, and the combined effect of socio-economic and natural factors far outweighed the influence of a single factor on carbon storage distribution. These findings can serve as a scientific reference and guide for mitigating carbon loss in ecosystems, promoting ecosystem protection, facilitating sustainable social development, and achieving the "dual carbon" goals in Zhejiang and other regions nationwide.

Personal Competence or Environmental Characteristics? Factors Influencing Happiness among Ageing Adults in Hong Kong Around the Period of COVID-19 Pandemic.

Drawing upon person-environment fit perspective, this study examines the joint and interactive influences of personal competence and environmental characteristics on the happiness of ageing adults around the period of COVID-19 pandemic. Data was collected in two rounds, before and during the pandemic, with 2,028 participants aged 55 years and older in Hong Kong. Personal competence encompassed financial status, physical health, and mental capital, while environmental characteristics included experiences of ageism, perceived social conditions, and age-friendly policies. Ordinary least squares regression was used to examine personal and environmental influences on happiness. Results indicated a significant decline in happiness following the onset of COVID-19 pandemic. Mental capital was found to have the strongest positive influence on happiness, followed by physical health, financial status, and social conditions. Mental capital mitigated the negative relationship between experiences of ageism and happiness. Practical interventions are informed to improve the well-being of ageing adults during pandemic.

[The clinic research of the modified laparoscopic splenectomy for massive splenomegaly in the treatment of children with hematologic diseases].

OBJECTIVE: To summarize the experience and characteristics of the modified laparoscopic splenectomy for massive splenomegaly in the treatment of children with hematologic disease. METHODS: The clinical data of 30 cases of laparoscopic splenectomy for massive splenomegaly of children with hematologic disease from March 2007 to December 2011 was analyzed retrospectively. There were 18 male and 12 female patients, aging from 2 to 14 years. Primary disease included mediterranean anemia (17 cases), hereditary spherocytosis (4 cases) and idiopathic thrombocytopenic purpura (ITP, 9 cases). Dissection started with cutting off the gastrosplenic ligaments and lesser sac to fully reveal the splenic hilum, the splenic artery was clamped twice with 10 mm tiatanum clamp. When most of blood stored in the spleen back to heart through the veins and the splenic volume had already decreased, the splenic vein was ligated with 10 mm titanium clip and cut with ligsure and splenic pedicle separated. The Surgery and complication were recorded. For 1 week after surgery, the hemoglobin and platelet counts were reviewed. RESULTS: Twenty-six cases were performed successfully, and 4 cases were converted to open procedure. Of the 4 cases, 2 cases was obesity because of idiopathic thrombocytopenic purpura, 1 case was ß thalassaemia combined severe liver enlargement, and 1 case was after partial splenic embolization. In cases of laparoscopic splenectomy, operation time was 110 to 130 minutes, with an average of 120 minutes, and blood loss during operation was 35 to 180 ml, with an average of 45 ml. Compared with pre-operation, the hemoglobin of mediterranean anemia and hereditary spherocytosis patients were (92 ± 8) g/L, and blood platelet count of ITP patients was (127 ± 20)×10(9)/L, and they increased obviously at 1 week after operation (t = 4.175 and 8.253, both P = 0.000). CONCLUSION: The modified surgical method make the laparoscopic splenectomy for massive splenomegaly in many children with hematologic diseases possible, which was thought to be impossible in the past.

Childhood Interpersonal Trauma and Depression of Middle-Aged and Older Adults in China: The Mediation Effect of Social Integration.

Objectives: This study investigates the associations between five types of childhood interpersonal trauma (CIT) and depression in mid-later life, as well as the mediation roles of social integration in the associations. Methods: Two waves of data from a nationally representative survey in China were used, with participants aged 45 years and older (N = 14,180). Measures of CIT included physical abuse, physical neglect, emotional neglect, bullying victimization, and exposure to interparental violence. Binary logistic regression and Karlson-Holm-Breen mediation analysis were used as statistical methods. Results: People who were bullied, physically abused, and witnessed interparental violence during childhood were more likely to develop depression in mid-later years. The associations between CIT and depression were mediated by satisfaction with marriage, satisfaction with children, and perceived social support. Discussion: Although CIT elevates the risk of depression in later life, effective interventions that strengthen integration across family and social domains could benefit mental health.

Experience of Chinese Recent Retirees on the Effects of Retirement on Healthy Ageing in Shenzhen and Hong Kong.

This study examined perspectives of recent retirees in Shenzhen and Hong Kong on how retirement influenced their healthy ageing. It investigated retirees' perceptions of healthy ageing and the ways in which healthy ageing connected with retirees' transition into retirement. A qualitative design with narrative interviews was used to interview twelve recent retirees in Shenzhen and thirteen in Hong Kong. The participants elaborated their perspectives on healthy ageing, which covered physical, mental, social, and financial domains. Retirees in both cities identified healthy ageing as maintaining an independent life and avoiding becoming a burden on family members. This study found that retirement declined physical health (in parallel with raised awareness of health promotion), posed both negative and positive influences on mental health, and shrank peripheral social networks of retirees. In addition, regional social welfare systems have different impacts on retirees' financial security and social participation. Retirees in Hong Kong reported higher stress of financial security and a strong desire for labor participation. Migrant-local welfare gaps were documented by retirees in Shenzhen. This study suggested that retirement planning, establishing a multi-pillar retirement-protection system, and narrowing the welfare gap between migrants and local residents should be implemented to reinforce healthy ageing.

Transfection of HCVc improves hTERT expression through STAT3 pathway by epigenetic regulation in Huh7 cells.

Previous studies showed that transient transfection of HCVc improved hTERT expression in hepatoma cell lines and it was noteworthy that phosphorylated signal transducer and activator of transcription 3 (pSTAT3) and DNA methyltransferases (DNMTs) were up regulated simultaneously. This study was designed to investigate the role of epigenetic regulation in the process of hTERT up regulation after HCVc transfection. Q-PCR and Western blot were used to analyze the expression of pSTAT3, DNMT1, and hTERT after the transfection of HCVc in hepatoma cell line Huh7. Proliferation and hTERT activity of Huh7 after HCVc transfection were examined by CCK8 and ELISA, respectively. Then, we blocked the JAK/STAT3 pathway or inhibited DNMT1 expression to investigate the regulation of pSTAT3, DNMT1, and hTERT. Methylation status of the promoter of hTERT gene was monitored by MS-PCR. Cell proliferation, hTERT expression level and activity of hTERT were promoted after HCVc transfection. The expression of pSTAT3 and DNMT1 were up-regulated simultaneously. DNMT1 and hTERT were down-regulated after blocking JAK/STAT3 pathway and the expression of hTERT weakened with DNMT1 inhibition. MS-PCR showed HCVc transfection increased the methylation level of hTERT promoter, and this effect was weakened after blocking the JAK/STAT3 pathway or with the treatment with DNMT1 inhibitor. HCVc transfection improved hTERT expression via epigenetic regulation. JAK/STAT3 pathway could be one of the essential factors in regulating DNMT1 expression during this process.

Selection of hepatocyte-like cells from mouse differentiated embryonic stem cells and application in therapeutic liver repopulation.

UNLABELLED: BACKGROUD/AIM: Because of the oncogenic risk, it is important to gain the homogeneous and purified cells from differentiated ESCs before transplantation. Here, we aim to select hepatocyte-like cells from differentiated ESCs, and investigate their growth, differentiation and neoplastic formation after intrahepatic transplantation. METHODS: Mouse ESCs were primarily induced by Dexamethesone, FGF-4 and HGF sequentially, then placed to a conditioning selection media consisting of 5% cholestatic sera and cultivated for 2 wks. After labeled by CFDA-SE, the selected cells were transplanted into mouse liver in therapeutic liver repopulation models. RESULTS: In the early stage of screening cultivation, most cells were suffered from apoptosis or even death. 1w later, some hepatocyte-like colony-forming units were observed, then the selected cells could grow and tend to be more mature, as assessed by morphological and functional tests. After intrahepatic transplantation, the labeled cells could proliferate and expressed albumin. Moreover, teratoma didn't form over 3 months. CONCLUSION: Our conditioning selection media could not only effectively select hepatocyte-like cells from differentiated ESCs, but further promote their growth and differentiation as well. After intrahepatic transplantation in therapeutic liver repopulation models, the selected cells could grow, differentiate and keep partial hepatic function. In particular, the transplantation was safe.

A modified laparoscopic splenectomy for massive splenomegaly in children with hematological disorder: a single institute retrospective clinical research.

PURPOSE: With the optimal acceptance of its clinical advantages, laparoscopic splenectomy (LS) emerged as a gold standard procedure as compared with open splenectomy (OS). However, it is still controversial and even counted as contraindication for massive splenomegaly. Here, we aim to summarize the experiences, characteristics and trends of modified LS for massive splenomegaly in children with hematological disorders. METHODS: Retrospective series of 57 pediatric patients with massive splenomegaly who underwent splenectomy from March 2007 to December 2011 were designated for this clinical analysis. The main outcome measures were dealt by statistics. For 30 cases of LS, we strictly adhered to the principle of using only three trocars to operate and initial ligation of the splenic artery, followed by retrieving the piecemeal of spleen through an accessory incision of 2-3 cm at 12 mm trocar port site. RESULTS: Of the 57 pediatric patients, 27 underwent OS and 30 underwent LS, respectively. Despite the operative time being shorter for OS than for LS (P < 0.001), the blood loss was lower in LS than in OS (P < 0.001); the time required for oral intake as well as duration of hospital stay was lower in LS than in OS (P < 0.001). Post-operatively, 7 (25.9 %) complications occurred in OS and 3 (10 %) in LS. The conversion rate of LS to OS was 13.33 % in four cases till 2009. CONCLUSIONS: Despite the conflicting reports regarding the safety of LS for massive splenomegaly, we demonstrated that our modified laparoscopic splenectomy in the treatment of children with massive splenomegaly in hematological diseases seemed to achieve the fundamental goal of less invasion; it was safe and feasible.

A Meta-Analysis on the Influence of Age-Friendly Environments on Older Adults' Physical and Mental Well-Being.

The importance of age-friendly environments (AFEs) for older adults has been empirically and theoretically highlighted by the extant literature. However, the strength of the association between environments and older adults' well-being has not been comprehensively quantified. Given the different attributes of the physical and mental dimensions, this meta-analysis aims to synthesise and quantify the association between AFEs and the physical and mental well-being of older adults. Fourteen eligible studies were included in this analysis: among which eight explored the link between AFEs and physical well-being, and eleven investigated AFEs in association with mental well-being. A random-effects model showed a small but significant correlation between AFEs and the mental well-being of older adults (r = 0.160, 95% CI [0.084, 0.224], p < 0.001), and the correlation between AFEs and physical well-being was also significant (r = 0.072, 95% CI [0.026, 0.118], p < 0.01). The number of environmental factors involved in AFEs moderated the association with physical well-being, from which the association was only significant among studies focusing on fewer environmental factors (n < 6). Results of this meta-analysis indicated that AFEs may be more effective in promoting the emotions of older adults, compared to ameliorating their physical functioning. The limitations of current empirical studies and directions for future research in the field of environmental gerontology were also discussed.

Saline short-term shock and rapid recovery on anammox performance.

Anaerobic ammonia oxidation (anammox) is an environmental-friendly biological nitrogen removal process, which has been developed as a promising technology in industrial wastewater treatment. However, anammox nitrogen removal under high saline conditions still faces many challenges. This study investigated the performance of anammox sludge under saline short-term shock and the strategy of rapid recovery. Salinity concentration, saline exposure time, and NaCl/Na2SO4 ratio were selected as three critical factors for short-term shock. The activity inhibition of anammox sludge were tested by using response surface methodology (RSM). Our results showed that, compared with the NaCl/Na2SO4 ratio, the salinity concentration and saline exposure time were the significant factor causing the anammox inhibition. The addition of glycine betaine (GB) in moderate amounts (0.1-5 mM) was found to help anammox to resist in relative low saline shock intensities (e.g., IC25 and IC50), with the activity retention rate of 94.7%. However, glycine betaine was not worked effectively under relatively high saline shock intensities (e.g., complete inhibition condition). Microbial community analysis revealed that Brocadiaceae accounted for only about 7.6%-13.2% at inhibited conditions. Interestingly, 16S rRNA analysis showed that the abundance of activated Brocadiaceae remarkably decreased with time after high-level saline shock. This tendency was consistent with the results of qPCR targeted hzsA gene. Finally, based on quorum sensing, the anammox activity was recovered to 93.5% of original sludge by adding 30% original sludge. The study realized the rapid recovery of anammox activity under complete inhibition, promoting the development and operation of salt-tolerant anammox process.

[Metagenomic and Metatranscriptomic Analysis of Nitrogen Removal Functional Microbial Community of Petrochemical Wastewater Biological Treatment Systems].

Petrochemicals are one of the pillar industries of China. Despite this, the treatment of petrochemical wastewater has long been seen as a massive challenge in the field of water pollution control, hindering the high-quality and sustainable development of the petrochemical industry. The majority of petrochemical enterprises and zones are located near rivers or seas, so their wastewater discharges can easily cause watershed or regional water ecological risks. Specifically, nitrogen pollution in petrochemical wastewater poses a significant threat to water ecological safety and human health. Sludge samples were collected from a petrochemical wastewater A/O nitrogen removal process line in a chemical industry zone in Shanghai. Metagenomic and metatranscriptomic methods were used to analyze the community structure of microorganisms, the functional characteristics of nitrogen removal bacteria, and the key nitrogen metabolism pathways in different sludges during the period when effluent water quality was stable and fluctuating. During the study, it was found that the nitrite and nitrate removal was relatively stable in this process, but ammonia oxidation fluctuated easily. In the study of microbial communities, it was found to be a nitrification-denitrification pathway that primarily removed nitrogen from the A/O process, and no genes related to ANAMMOX were detected. Approximately 90% of the functional genes responsible for removing nitrogen were responsible for denitrification, whereas only 0.17% of them were involved in the conversion of ammonia nitrogen in the nitrification process. Moreover, the abundance of ammonia-oxidizing bacteria in the process was extremely low, and the main genus was Nitrosomonas. It is likely that this is the main cause of fluctuations in ammonia nitrogen concentration in effluent due to water quality shocks in the process line.

Functional Differentiation of BMP7 Genes in Zebrafish: bmp7a for Dorsal-Ventral Pattern and bmp7b for Melanin Synthesis and Eye Development.

Bone morphogenetic protein 7 (BMP7) belongs to the transforming growth factor ß (TGF-ß) family, which not only induces cartilage and bone formation, but also regulates eye development and melanoma tumorigenesis in mammals. In teleosts, BMP7 differentiates into two subtypes, bmp7a and bmp7b, which have clearly differentiated structures. To fully understand the functional differentiation of bmp7a and bmp7b in fish species, we successfully constructed bmp7a and bmp7b gene deletion mutants in zebrafish using CRISPR/Cas9-mediated gene editing technology. Our results showed that bmp7a mutation caused abnormal development of the embryo's dorsal-ventral pattern that led to death; bmp7b mutation induced growth inhibition and increased melanin production in the skin and eye of mutants. Histological analysis revealed that melanin in the retina of the eyes in bmp7b mutants increased, and behavioral observation showed that the vision and sensitivity to food of the mutants were reduced. Transcriptome analysis of the skin and eye tissues showed that the expression changes of wnt7ba and gna14 in bmp7b mutants might promote the increase of melanin. Additionally, the eye transcriptome analysis indicated that changes in the structure of the eyes in bmp7b mutants led to defects in phototransduction, and seven DEGs (rgs9a, rgs9b, rcvrn2, guca1d, grk1b, opn1mw4, and gc2) were identified as key candidate genes that affected the photonic response of the eyes. The study revealed the functional differentiation of bmp7a and bmp7b in teleosts and the first report about the inhibitory effect of bmp7b on melanogenesis may provide useful information for the future research on human melanoma-related diseases.

Single-cell transcriptomes and runx2b-/- mutants reveal the genetic signatures of intermuscular bone formation in zebrafish.

Intermuscular bones (IBs) are mineralized spicules, present in the myosepta of many, but not all, teleost species. IBs are often small and sharp, and they consequently limit how the fish can be processed; the IBs may cause injury or trauma if lodged in consumers' throats or mouths, and therefore affect the appeal of the fish to many consumers. The development of IBs in teleosts is still not fully understood and the molecular basis of IB development remains to be established. Here, the characteristics of IB tissue are evaluated based on single-cell transcriptomics in wild-type zebrafish. The analysis defined 18 distinct cell types. Differentiation trajectories showed that IBs are derived from tendons and that a core tendon-osteoblast cell lineage is related to IB formation. In particular, the functions of 10 candidate genes were evaluated via CRISPR-Cas9 mutants. Among those, runx2b-/- mutants completely lost IBs, while swimming performance, growth and bone mineral density were not significantly different from runx2b+/+ zebrafish. Comparative single-cell RNA sequencing (scRNA-seq) analysis in runx2b-/- and runx2b+/+ zebrafish revealed the role of osteoblasts in IB formation. In addition, differentially expressed genes were enriched in the transforming growth factor ß/bone morphogenetic protein (TGF-ß/BMP) pathway after runx2b deletion. This study provides evidence for the crucial role of runx2b regulation in IB formation. Genetic breeding can target runx2b regulation and generate strains of commercial fish species without IBs, which can improve the safe consumption and economic value of many farmed fish species.

The anammox coupled partial-denitrification process in an integrated granular sludge and fixed-biofilm reactor developed for mainstream wastewater treatment: Performance and community structure.

This study describes an integrated granular sludge and fixed-biofilm (iGB) reactor innovatively designed to carry out the anammox/partial-denitrification (A/PD) process for nitrogen removal with mainstream municipal wastewater. The iGB-A/PD reactor consists of anammox granules inoculated in the lower region of reactor and an acclimated fixed-biofilm positioned in the upper region. Compared to the other reported A/PD systems for mainstream wastewater treatment, this iGB-A/PD reactor is notable due to its higher quality effluent with a total inorganic nitrogen (TIN) of ∼3 mg•L-1 and operation at a high nitrogen removal rate (NRR) of 0.8 ± 0.1 kg-N•m-3•d-1. Reads-based metatranscriptomic analysis found that the expression values of hzsA and hdh, key genes associated with anammox, were much higher than other functional genes on nitrogen conversion, confirming the major roles of the anammox bacteria in nitrogen bio-removal. In both regions of the reactor, the nitrate reduction genes (napA/narG) had expression values of 56-99 RPM, which were similar to that of the nitrite reduction genes (nirS/nirK). The expression reads from genes for dissimilatory nitrate reduction to ammonium (DNRA), nrfA and nirB, were unexpectedly high, and were over the half of the levels of reads from genes required for nitrate reduction. Kinetic assays confirmed that the granules had an anammox activity of 16.2 g-NH4+-N•kg-1-VSS•d-1 and a nitrate reduction activity of 4.1 g-N•kg-1-VSS•d-1. While these values were changed to be 4.9 g- NH4+-N•kg-1-VSS•d-1and 4.3 g-N•kg-1-VSS•d-1 respectively in the fixed-biofilm. Mass flux determination found that PD and DNRA was responsible for ∼50% and ∼25% of nitrate reduction, respectively, in the whole reactor, consistent with high effluent quality and treatment efficiency via a nitrite loop. Metagenomic binning analysis revealed that new and unidentified anammox species, affiliated with Candidatus Brocadia, were the dominant anammox organisms. Myxococcota and Planctomycetota were the principal organisms associated with the PD and DNRA processes, respectively.

Comparison of Myosepta Development and Transcriptome Profiling between Blunt Snout Bream with and Tilapia without Intermuscular Bones.

Intermuscular bones (IBs) are small spicule-like bones located in the myosepta of basal teleosts, which negatively affect the edibleness and economic value of fish. Blunt snout bream (Megalobrama amblycephala, with epineural and epipleural IBs) and tilapia (Oreochromis niloticus, without epineural and epipleural IBs) are two major aquaculture species and ideal models for studying the formation mechanisms of fish IBs. Here, we compared myosepta development between M. amblycephala and O. niloticus, based on histological analysis, transcriptome profiling, and expression analysis of bone-related genes. The histological results showed that dye condensation began to appear in the myosepta 20 days post hatching (dph) in M. amblycephala, and IBs could be clearly observed 50 dph in the myosepta, based on different staining methods. However, in O. niloticus, dye condensation was not observed in the myosepta from 10 to 60 dph. Differentially expressed genes (DEGs) at different developmental stages were screened by comparing the transcriptomes of M. amblycephala and O. niloticus, and KEGG analysis demonstrated that these DEGs were enriched in many bone-related pathways, such as focal adhesion, calcium, and Wnt signaling pathways. Quantitative PCR was performed to further compare the expression levels of some bone-related genes (scxa, scxb, runx2a, runx2b, bgp, sp7, col1a2, entpd5a, entpd5b, phex, alpl, and fgf23). All the tested genes (except for alpl) exhibited higher expression levels in M. amblycephala than in O. niloticus. A comparison of the dorsal and abdominal muscle tissues between the two species also revealed significant expression differences for most of the tested genes. The results suggest that scxa, scxb, runx2a, runx2b, entpd5a, col1a2, and bgp may play important roles in IB development. Our findings provide some insights into the molecular mechanisms of IB formation, as well as clues for further functional analysis of the identified genes to better understand the development of IBs.

[Synthesis of colon-specific prodrug of indomethacin and its inhibitory effect on liver metastasis from colon cancer].

OBJECTIVE: To develop a colon-specific prodrug of Indomethacin microbially triggered, carry out in vitro/in vivo evaluation of drug release, and appraise its inhibitory effect on liver metastasis from colon cancer. METHODS: Indomethacin prodrugs were synthesized and characterized by FTIR and NMR, and dissolution test simulating gastrointestinal tract was employed to screen the colon-specific prodrug. Then, the pharmacokinetic profile of portal vein and peripheral blood in Sprague-Dawley rats was studied. Lastly, the inhibitory effect on liver metastasis from colon cancer in nude mice was observed. RESULTS: The chemical structure characterized by FTIR and NMR demonstrated that six kinds of indomethacin-block-amylose with different drug loading (IDM-AM-1-6) were synthesized, among which IDM-AM-3 was degraded 1.3%, 9.3% and 95.3%, respectively, in simulated gastric fluid for 4 h, small intestine for 6 h, and colon for 36 h. The pharmacokinetic test of IDM-AM-3 showed that absorption was delayed significantly (P < 0.01), peak time [(11.35 + or - 2.45) h], elimination half-life [(16.74 + or - 4.04) h] and mean residence time [(22.27 + or - 0.52) h] were significantly prolonged (P < 0.01), as well as peak serum concentrations [(9.69 + or - 2.40) mg/L] and AUC(0-t) [(236.7 + or - 13.1) mg x L(-1) x h] were decreased markedly (P < 0.01) as compared with those of IDM regarding to portal vein. Additionally, its AUC(0-t) in peripheral blood was remarkably lower than that in Portal vein (P < 0.01). The tumor suppression observation showed that it could remarkably reduce the number of liver metastases in contrast to IDM (P < 0.05). CONCLUSION: Colon-specific IDM-AM-3 possesses advantage of sustained release in portal vein providing some experimental basis for colon-specific delivery system applied to sustained release in the portal vein.

Preparation and antitumor effects of nanovaccines with MAGE-3 peptides in transplanted gastric cancer in mice.

BACKGROUND AND OBJECTIVE: As a prospective vaccine carrier, nanoparticles can protect antigens from degradation and enhance immune response. This study prepared nanovaccines with MAGE-3-derived CD4+-CD8+T cell epitope peptides, and investigated its character and antitumor effects on transplanted gastric cancer in mice. METHODS: We adopted the self-assembly method to prepare peptide/chitosan conjugated with deoxycholic acid (chitosan-deoxycholic acid) nanoparticles. We observed the appearance of the chitosan-deoxycholic acidnanoparticles through a transmission electron microscope (TEM) and analyzed the peptide content and its release pattern by fluorescence spectrophotometry. We observed tumor-suppression efficacy in vivo through animal experiments. RESULTS: We successfully prepared nanoparticles with MAGE-3 peptide antigen, and its encapsulation efficiency and loading level were about 37% and 17.0%, respectively. These nanoparticles presented a delayed release pattern in phosphate buffered saline (PBS) at pH 7.4, and the full release time was about 48 h. In 2 mg/mL lysozyme, the nanoparticles showed a sudden release, and the full release time was about 24 h. ELISPOT and cytotoxic experiments showed that the MAGE-3 peptide loaded nanoparticles could stimulate immune response in vivo and could generate MAGE-3-targeted cytotoxic T lymphocytes (CTLs), and kill MAGE-3-specific tumor cells. Tumor suppression experiments showed that the regression ratio of the peptide-loaded nanoparticles group was 37.81%. CONCLUSIONS: MAGE-3 peptide/chitosan-deoxycholic acidvaccine-loaded nanoparticles can stimulate antitumor immune response in vivo and can regress the growth of mouse forestomach carcinoma cell line MFC.