Metabolic risks remain a serious threat to cardiovascular disease: findings from the Global Burden of Disease Study 2019.

Metabolic factors are major and controllable risk factors for cardiovascular diseases (CVD), and few studies have described this burden. We aim to assess it from 1990 to 2019 and predict the trends through 2034. Global Burden of Disease (GBD) provides data on sex, age, and socio-demographic index (SDI) levels. Numbers, age-standardized death rates (ASDR) and estimated annual percentage change (EAPC) were used. Future trends were estimated by NORDPRED model. The deaths cases of metabolic-related CVD increased from 8.61 million (95% UI: 7.91-9.29) to 13.71 million (95% UI: 12.24-14.94) globally. The ASDR continued to decline globally (EAPC = -1.36). The burden was heavier in male and middle-aged people and elderly people. CVD-related ASDR caused by high systolic blood pressure (SBP) had a downward trend globally (EAPC = -1.45), while trends of high body mass index (BMI) (EAPC = 1.29, 1.97, 0.92) and fasting plasma glucose (FPG) (EAPC = 0.95, 1.08, 0.46) were increasing in the middle, low-middle, and low SDI regions, respectively. Compared to 2015-2019, cumulative deaths will increase by 27.85% from 2030 to 2034, while ASDR will decrease 10.47%. The metabolic-related CVD burden remained high globally and deaths will continue to rise in the future. Men, middle-aged and elderly people were focus of concern. High SBP was globally well-managed over the past 30 years, but the CVD burden due to high BMI and FPG remained high. Exceptional initiatives are needed to regarding interventions targeting high BMI and FPG in middle and lower SDI regions.

Acute kidney injury in patients treated with immune checkpoint inhibitors: a single-center retrospective study.

BACKGROUND: Immune checkpoint inhibitor-associated acute kidney injury (ICI-AKI) is the most common renal complication and has attracted increasing amounts of attention. However, studies on this topic in Chinese cancer patients are very limited. Therefore, we conducted a retrospective study on the incidence, risk factors, clinical features and renal recovery of ICI-AKI in all patients with malignancies treated with ICIs in Shandong Provincial Hospital Affiliated to Shandong First Medical University. METHODS: In this single-center retrospective cohort study, the data of 904 patients who received immune checkpoint inhibitors (ICIs) treatment were retrospectively analyzed. Multivariable logistic regression was used to identify the predictors of ICI-AKI. RESULTS: A total of 46 of 904 patients receiving ICIs developed ICI-AKI, and the incidence of ICI-AKI was 5.1%. Patients developed ICI-AKI at a median of 9 weeks (IQR 3-23) after ICIs initiation. A lower baseline estimated glomerular filtration rate (eGFR) and use of antibiotics were associated with a higher risk of ICI-AKI. Renal recovery occurred in 17 patients (46%) at a median of 4 weeks (IQR 2-8) after ICI-AKI, including 16 (43%) with complete recovery and 1 (3%) with partial recovery. Of the 14 rechallenged patients, only one developed recurrent ICI-AKI. CONCLUSIONS: Patients with ICI-AKI were more likely to have impaired renal function at baseline and after treatment with antibiotics. Approximately half of the patients achieved renal recovery.

Association of macronutrient consumption quality, food source and timing with depression among US adults: A cross-sectional study.

BACKGROUND: Growing evidence suggests that meal timing may influence dietary choices and mental health. Thus, this study examined the association between macronutrient consumption quality, food source, meal timing, and depression prevalence in Americans. METHODS: 23,313 National Health and Nutrition Survey participants from 2007 to 2016 were included in this cross-sectional study. Macronutrient intake was calculated for all day, dinner, and breakfast and subtypes into 4 classes. Based on the Patient Health Questionnaire, depression was defined as a 9-item score ≥ 10 on the PHQ-9. The correlation between macronutrients and depression prevalence was estimated with multivariable logistic regression models and isocaloric substitution effects. RESULTS: Low-quality carbohydrates (OR = 1.54, 95 % CI: 1.11, 2.12) were positively linked to depression compared with the lowest quartile, after adjusting for age and other covariates. In contrast, total high-quality carbohydrate (OR = 0.52, 95 % CI: 0.40, 0.66), total animal protein (OR = 0.60, 95 % CI: 0.45, 0.80), and total vegetable protein (OR = 0.61, 95 % CI: 0.43, 0.85) were negatively associated with depression was negatively associated. Replacing low-quality carbohydrates with high-quality carbohydrates throughout the day reduced the risk of depression by approximately 15 %. LIMITATIONS: Cross-sectional data. CONCLUSION: All in all, diet plays a crucial role in the prevention and treatment of depression. Especially in terms of macronutrient intake, high-quality, moderate intake can reduce the risk of depression. However, different subtypes of macronutrient consumption may have different effects on depression, so it becomes crucial to carefully consider the selection and combination of macronutrients.

Association between inflammatory biomarkers and mortality in individuals with type 2 diabetes: NHANES 2005-2018.

PURPOSE: This study aimed to examine independent association between inflammatory biomarkers and all-cause mortality as well as cardio-cerebrovascular disease (CCD) mortality among U.S. adults with diabetes. METHODS: A cohort of 6412 U.S. adults aged 20 or older was followed from the start until December 31, 2019. Statistical models such as Cox proportional hazards model (Cox) and Kaplan-Meier (K-M) survival curves were employed to investigate the associations between the inflammatory biomarkers and all-cause mortality and CCD mortality. RESULTS: After adjusting for confounding factors, the highest quartile of inflammatory biomarkers (NLR HR = 1.99; 95 % CI:1.54-2.57, MLR HR = 1.93; 95 % CI:1.46-2.54, SII HR = 1.49; 95 % CI:1.18-1.87, SIRI HR = 2.32; 95 % CI:1.81-2.96, nLPR HR = 2.05; 95 % CI:1.61-2.60, dNLR HR = 1.94; 95 % CI:1.51-2.49, AISI HR = 1.73; 95 % CI:1.4 1-2.12)) were positively associated with all-cause mortality compared to those in the lowest quartile. K-M survival curves indicated that participants with an inflammatory biomarker above a certain threshold had a higher risk of both all-cause mortality and CCD mortality (Log rank P < 0.05). CONCLUSION: Some biomarkers such as NLR, MLR, SII, AISI, SIRI, and dNLR, are significantly associated with all-cause mortality and CCD mortality among U.S. adults with diabetes. The risk of both outcomes increased when the biomarkers surpassed a specific threshold.

A DNA/Upconversion Nanoparticle Complex Enables Controlled Co-Delivery of CRISPR-Cas9 and Photodynamic Agents for Synergistic Cancer Therapy.

Photodynamic therapy (PDT) depends on the light-irradiated exciting of photosensitizer (PS) to generate reactive oxygen species (ROS), which faces challenges and limitations in hypoxia and antioxidant response of cancer cells, and limited tissue-penetration of light. Herein, a multifunctional DNA/upconversion nanoparticles (UCNPs) complex is developed which enables controlled co-delivery of CRISPR-Cas9, hemin, and protoporphyrin (PP) for synergistic PDT. An ultralong single-stranded DNA (ssDNA) is prepared via rolling circle amplification (RCA), which contains recognition sequences of single guide RNA (sgRNA) for loading Cas9 ribonucleoprotein (RNP), G-quadruplex sequences for loading hemin and PP, and linker sequences for combining UCNP. Cas9 RNP cleaves the antioxidant regulator nuclear factor E2-related factor 2 (Nrf2), improving the sensitivity of cancer cells to ROS, and enhancing the synergistic PDT effect. The G-quadruplex/hemin DNAzyme mimicks horseradish peroxidase (HRP) to catalyze the endogenous H2O2 to O2, overcoming hypoxia condition in tumors. The introduced UCNP converts NIR irradiation with deep tissue penetration to light with shorter wavelength, exciting PP to transform the abundant O2 to 1O2. The integration of gene editing and PDT allows substantial accumulation of 1O2 in cancer cells for enhanced cell apoptosis, and this synergistic PDT has shown remarkable therapeutic efficacy in a breast cancer mouse model.

FTO-mediated m6A modification of serum amyloid A2 mRNA promotes podocyte injury and inflammation by activating the NF-κB signaling pathway.

Diabetic kidney disease (DKD) is one of the severe complications of diabetes mellitus, yet there is no effective treatment. Exploring the development of DKD is essential to treatment. Podocyte injury and inflammation are closely related to the development of DKD. However, the mechanism of podocyte injury and progression in DKD remains largely unclear. Here, we observed that FTO expression was significantly upregulated in high glucose-induced podocytes and that overexpression of FTO promoted podocyte injury and inflammation. By performing RNA-seq and MeRIP-seq with control podocytes and high glucose-induced podocytes with or without FTO knockdown, we revealed that serum amyloid A2 (SAA2) is a target of FTO-mediated m6A modification. Knockdown of FTO markedly increased SAA2 mRNA m6A modification and decreased SAA2 mRNA expression. Mechanistically, we demonstrated that SAA2 might participate in podocyte injury and inflammation through activation of the NF-κB signaling pathway. Furthermore, by generating podocyte-specific adeno-associated virus 9 (AAV9) to knockdown SAA2 in mice, we discovered that the depletion of SAA2 significantly restored podocyte injury and inflammation. Together, our results suggested that upregulation of SAA2 promoted podocyte injury through m6A-dependent regulation, thus suggesting that SAA2 may be a therapeutic target for diabetic kidney disease.

Age-dependent interaction between Life's Essential 8 and chronic kidney disease: A national cross-sectional analysis.

OBJECTIVE: Chronic kidney disease (CKD), an age-related condition, is closely associated with cardiovascular disease. We aimed to examine the age-dependent interaction between Life's Essential 8 (LE8), the updated measurement of cardiovascular health (CVH), and CKD in the United States. METHODS: The cross-sectional study involved 25,529 participants from National Health and Nutrition Examination Survey in 2007-2018. Multivariate logistic regressions were used to estimate the age-dependent interaction between LE8 and CKD, and restricted cubic spline regressions were used to analyze the dose-response relationships between LE8 and CKD among adults and all age subgroups. RESULTS: Overall, 2934 (9.3%), 17,278 (66.2%), and 5317 (24.5%) participants had low, moderate, and high CVH, separately. After adjusting for the potential covariates, LE8 was negatively associated with CKD [odds ratio (OR) for per 1 standard deviation (SD) increase and 95%CI, 0.71 (0.67, 0.75)], with a nonlinear dose-response relationship (P for nonlinearity = 0.001). The inversed association was stronger among participants aged 65 and older (0.65 (0.59, 0.71)) compared to youngers [20-39 years, 0.63 (0.59, 0.58), 40-64 years, 0.63 (0.59, 0.58)] (P for interaction = 0.002). CONCLUSIONS: CVH, as measured by the LE8 score, was negatively associated with the presence of CKD in non-linear fashions, more pronounced in participants aged 65 and older compared to younger age groups.

N6-methyladenosine methylation in kidney injury.

Multiple mechanisms are involved in kidney damage, among which the role of epigenetic modifications in the occurrence and development of kidney diseases is constantly being revealed. However, N6-methyladenosine (M6A), a well-known post-transcriptional modification, has been regarded as the most prevalent epigenetic modifications in higher eukaryotic, which is involved in various biological processes of cells such as maintaining the stability of mRNA. The role of M6A modification in the mechanism of kidney damage has attracted widespread attention. In this review, we mainly summarize the role of M6A modification in the progression of kidney diseases from the following aspects: the regulatory pattern of N6-methyladenosine, the critical roles of N6-methyladenosine in chronic kidney disease, acute kidney injury and renal cell carcinoma, and then reveal its potential significance in the diagnosis and treatment of various kidney diseases. A better understanding of this field will be helpful for future research and clinical treatment of kidney diseases.

Content-Seam-Preserving Multi-Alignment Network for Visual-Sensor-Based Image Stitching.

As an important representation of scenes in virtual reality and augmented reality, image stitching aims to generate a panoramic image with a natural field-of-view by stitching multiple images together, which are captured by different visual sensors. Existing deep-learning-based methods for image stitching only conduct a single deep homography to perform image alignment, which may produce inevitable alignment distortions. To address this issue, we propose a content-seam-preserving multi-alignment network (CSPM-Net) for visual-sensor-based image stitching, which could preserve the image content consistency and avoid seam distortions simultaneously. Firstly, a content-preserving deep homography estimation was designed to pre-align the input image pairs and reduce the content inconsistency. Secondly, an edge-assisted mesh warping was conducted to further align the image pairs, where the edge information is introduced to eliminate seam artifacts. Finally, in order to predict the final stitched image accurately, a content consistency loss was designed to preserve the geometric structure of overlapping regions between image pairs, and a seam smoothness loss is proposed to eliminate the edge distortions of image boundaries. Experimental results demonstrated that the proposed image-stitching method can provide favorable stitching results for visual-sensor-based images and outperform other state-of-the-art methods.

Cascade dynamic assembly/disassembly of DNA nanoframework enabling the controlled delivery of CRISPR-Cas9 system.

CRISPR-Cas9 has been explored as a therapeutic agent for down-regulating target genes; the controlled delivery of Cas9 ribonucleoprotein (RNP) is essential for therapeutic efficacy and remains a challenge. Here, we report cascade dynamic assembly/disassembly of DNA nanoframework (NF) that enables the controlled delivery of Cas9 RNP. NF was prepared with acrylamide-modified DNA that initiated cascade hybridization chain reaction (HCR). Through an HCR, single-guide RNA was incorporated to NF; simultaneously, the internal space of NF was expanded, facilitating the loading of Cas9 protein. NF was designed with hydrophilic acylamino and hydrophobic isopropyl, allowing dynamic swelling and aggregation. The responsive release of Cas9 RNP was realized by introducing disulfide bond-containing N,N-bis(acryloyl)cystamine that was specifically in response to glutathione of cancer cells, triggering the complete disassembly of NF. In vitro and in vivo investigations demonstrated the high gene editing efficiency in cancer cells, the hypotoxicity in normal cells, and notable antitumor efficacy in a breast cancer mouse model.

Association of perfluoroalkyl and polyfluoroalkyl substances with sex hormones in children and adolescents 6-19 Years of age.

Perfluoroalkyl and polyfluoroalkyl substances (PFAS) have raised concerns regarding sex hormones homeostasis disruption in critical windows including childhood and adolescence, but epidemiological evidence is limited. We aimed to explore the associations of total testosterone (TT), estradiol (E2), and sex hormone binding globulin (SHBG) in children and adolescents with PFAS in 921 participants 6-19 years of age from NHANES 2013 to 2016. Multiple linear regression models and Bayesian Kernel Machine Regression (BKMR) models stratified by sex-age and sex-puberty-status groups were performed to explore the associations of the associations of individual or mixture of PFAS with sex hormone levels, respectively. Inverse associations were observed between n-PFOA and SHBG in female adolescents when the exposure was modeled as continuous (ß = -0.20, 95% CI -0.33, -0.07) or categorized variable (P for trend = 0.005). In children, inverse associations were observed by BKMR in 6-11-year-old girls of high concentration, and in boys of low concentration of the PFAS mixture with TT. A positive association of PFAS mixture with SHBG was observed in boys. PFOS and PFNA were identified as major contributors to the associations in girls and boys, respectively. Although the 95% credible intervals included the null in adolescents, suggestive negative associations of PFAS mixture with TT and SHBG levels in adolescents aged 12-19 years were found by BKMR. Results by sex-puberty status presented a similar pattern, where significantly inverse associations between PFAS mixture and E2 were observed in the pubertal. Our findings suggested the associations of either individual or mixture PFAS with decreased TT levels, and increased SHBG levels in U.S. children and adolescents, and with decreased E2 levels in pubertal individuals. The associations were evident in children.

ROS promote hyper-methylation of NDRG2 promoters in a DNMTS-dependent manner: Contributes to the progression of renal fibrosis.

Renal fibrosis is the common histopathological feature of chronic kidney diseases (CKD), and there is increasing evidence that epigenetic regulation is involved in the occurrence and progression of renal fibrosis. N-myc downstream-regulated gene 2 (NDRG2) is significantly down-regulated in renal fibrosis, the mechanism of which remains unclear. Previous studies have confirmed that the inhibition of NDRG2 expression in tumor cells is related to hyper-methylation, mainly regulated by DNA methyltransferases (DNMTS). Herein, we explored the expression of NDRG2 and its epigenetic regulatory mechanism in renal fibrosis. The results showed that the expression of NDRG2 was significantly inhibited in vivo and in vitro, while the overexpression of NDRG2 effectively alleviated renal fibrosis. Meanwhile, we found that the expression of DNMT1/3A/3B was significantly increased in hypoxia-induced HK2 cells and Unilateral Ureteral Obstruction (UUO) mice accompanied by hyper-methylation of the NDGR2 promoter. Methyltransferase inhibitor (5-AZA-dC) corrected the abnormal expression of DNMT1/3A/3B, reduced the methylation level of NDRG2 promoter and restored the expression of NDRG2. The upstream events that mediate changes in NDRG2 methylation were further explored. Reactive oxygen species (ROS) are important epigenetic regulators and have been shown to play a key role in renal injury due to various causes. Accordingly, we further explored whether ROS could induce DNA-epigenetic changes of the expression of NDRG2 and then participated in the development of renal fibrosis. Our results showed that mitochondria-targeted antioxidants (Mito-TEMPO) could reverse the epigenetic inhibition of NDRG2 in a DNMT-sensitive manner, showing strong ability of DNA demethylation, exhibiting epigenetic regulation and anti-fibrosis effects similar to 5-AZA-dC. More importantly, the anti-fibrotic effects of 5-AZA-dC and Mito-TEMPO were eliminated in HK2 cells with NDRG2 knockdown. These findings highlight that targeting ROS-mediated hyper-methylation of NDRG2 promoter is a potentially effective therapeutic strategy for renal fibrosis, which will provide new insights into the treatment of CKD.

Long noncoding RNA ENST00000436340 promotes podocyte injury in diabetic kidney disease by facilitating the association of PTBP1 with RAB3B.

Dysfunction of podocytes has been regarded as an important early pathologic characteristic of diabetic kidney disease (DKD), but the regulatory role of long noncoding RNAs (lncRNAs) in this process remains largely unknown. Here, we performed RNA sequencing in kidney tissues isolated from DKD patients and nondiabetic renal cancer patients undergoing surgical resection and discovered that the novel lncRNA ENST00000436340 was upregulated in DKD patients and high glucose-induced podocytes, and we showed a significant correlation between ENST00000436340 and kidney injury. Gain- and loss-of-function experiments showed that silencing ENST00000436340 alleviated high glucose-induced podocyte injury and cytoskeleton rearrangement. Mechanistically, we showed that fat mass and obesity- associate gene (FTO)-mediated m6A induced the upregulation of ENST00000436340. ENST00000436340 interacted with polypyrimidine tract binding protein 1 (PTBP1) and augmented PTBP1 binding to RAB3B mRNA, promoted RAB3B mRNA degradation, and thereby caused cytoskeleton rearrangement and inhibition of GLUT4 translocation to the plasma membrane, leading to podocyte injury and DKD progression. Together, our results suggested that upregulation of ENST00000436340 could promote podocyte injury through PTBP1-dependent RAB3B regulation, thus suggesting a novel form of lncRNA-mediated epigenetic regulation of podocytes that contributes to the pathogenesis of DKD.

Plant cultivar determined bacterial community and potential risk of antibiotic resistance gene spread in the phyllosphere.

The global increased antibiotic resistance level in pathogenic microbes has posed a significant threat to human health. Fresh vegetables have been recognized to be an important vehicle of antibiotic resistance genes (ARGs) from environments to human beings. Phyllosphere ARGs have been indicated to be changed with plant species, yet the influence of plant cultivar on the phyllospheric resistome is still unclear. Here, we detected the ARGs and bacterial communities in the phyllosphere of two cultivars of cilantros and their corresponding soils using high-throughput quantitative PCR technique and bacterial 16S rRNA gene-based high-throughput sequencing, respectively. We further identified the potential bacterial pathogens and analyzed the effects of plant cultivar on ARGs, mobile genetic elements (MGEs), microbiome and potential bacterial pathogens. The results showed that the cultivars did not affect the ARG abundance and composition, but significantly shaped the abundance of MGEs and the composition structure of bacteria in the phyllosphere. The relative abundance of potential bacterial pathogens was significantly higher in the phyllosphere than that in soils. Mantel test showed that the ARG patterns were significantly correlated to the patterns of potential bacterial pathogens. Our results suggested that the horizontal gene transfer of ARGs in the phyllosphere might be different between the two cultivars of cilantro and highlighted the higher risk of phyllospheric microorganisms compared with those in soils. These findings extend our knowledge on the vegetable microbiomes, ARGs, and potential pathogens, suggesting more agricultural and hygiene protocols are needed to control the risk of foodborne ARGs.

Underestimation about the Contribution of Nitrate Reducers to Iron Cycling Indicated by Enterobacter Strain.

Nitrate-reducing iron(II) oxidation (NRFO) has been intensively reported in various bacteria. Iron(II) oxidation is found to be involved in both enzymatic and chemical reactions in nitrate-reducing Fe(II)-oxidizing microorganisms (NRFOMs). However, little is known about the relative contribution of biotic and abiotic reactions to iron(II) oxidation for the common nitrate reducers during the NRFO process. In this study, the typical nitrate reducers, four Enterobacter strains E. hormaechei, E. tabaci, E. mori and E. asburiae, were utilized as the model microorganisms. The comparison of the kinetics of nitrate, iron(II) and nitrite and N2O production in setups with and without iron(II) indicates a mixture of enzymatic and abiotic oxidation of iron(II) in all four Enterobacter strains. It was estimated that 22-29% of total oxidized iron(II) was coupled to microbial nitrate reduction by E. hormaechei, E. tabaci, E. mori, and E. asburiae. Enterobacter strains displayed an metabolic inactivity with heavy iron(III) encrustation on the cell surface in the NRFOmedium during days of incubation. Moreover, both respiratory and periplasmic nitrate-reducing genes are encoded by genomes of Enterobacter strains, suggesting that cell encrustation may occur with periplasmic iron(III) oxide precipitation as well as the surface iron(II) mineral coating for nitrate reducers. Overall, this study clarified the potential role of nitrate reducers in the biochemical cycling of iron under anoxic conditions, in turn, re-shaping their activity during denitrification because of cell encrustation with iron(III) minerals.

Physical and Chemical Properties of Coal Gasification Fine Slag and Its Carbon Products by Hydrophobic-Hydrophilic Separation.

Coal gasification fine slag is a kind of solid waste with low resource utilization rate. The complex embedding of residual carbon and inorganic minerals (ash materials) is the main reason restricting the efficient resource separation and utilization of residual carbon or ash materials. Hydrophobic-hydrophilic separation (HHS) is a separation technology in which mineral particles with different surface hydrophobicity values are enriched in the water phase and oil phase under the action of mechanical stirring. The water on the surface of hydrophobic particles is replaced by the oil phase to form flocs, which are enriched in the hydrophobic liquid phase, while hydrophilic particles are dispersed into the aqueous phase. In this study, the HHS process was used to separate the carbon/ash from the fine gasification slag produced by a Shenning gasifier, Texaco gasifier, and GSP gasifier of Ningxia Coal Industry Co., Ltd. The physicochemical properties of the original sample and the residual carbon products obtained by hydrophobic-hydrophilic separation were analyzed. The results show that HHS can separate the carbon/ash in the three kinds of fine slag to varying degrees. The carbon element is enriched into the hydrophobic phase to form the concentrates, while the silicon element, oxygen element, and metal element enter the tailings. The spherical ash with different particle sizes distributed on the surface of residual carbon and the gap of the matrix is basically removed, while the ash in the carbon-ash melt is difficult to remove. The ash contents of the concentrate and tailings of fine slag of the Shenning gasifier are 22.58 and 96.28%, respectively, which reach the best ash index compared with that of the other two gasifiers. From the change of mineral surface properties after HHS, the distribution of oxygen-containing groups, benzene rings, Si-O, and clay minerals or carbonate minerals in the three kinds of fine slag residual carbon products is basically similar. Compared with the other two gasifier products, the GSP gasifier concentrate has a larger specific surface area and less ash material, more amorphous carbon structures (less graphitic), and more active sites, resulting in a stronger combustion activity.

[Theory of "synchronization of heaven-qi, earth-qi and human-qi" in Zhuang Medicine and its application in acupuncture and moxibustion].

The connotation of theory of "synchronization of heaven-qi, earth-qi and human-qi " in Zhuang Medicine is introduced, and its association with human physiological state, pathological changes and acupuncture treatment mechanism are discussed. The main points, acupuncture manipulation and clinical acupoint selection of acupuncture and moxibustion of Zhuang Medicine are guided by this theory. The points are mainly composed of Huan points, Luoyang points and experience-based points, which are divided into heaven part, earth part and human part according to the positions in the human body. The acupuncture manipulation is divided into mild manipulation, moderate manipulation and heavy manipulation according to heaven, earth and human theory. With the point prescriptions for headache, dysmenorrhea and herpes zoster sore as examples, the application of this theory in the clinical acupoint selection is explained.

Host age increased conjugal plasmid transfer in gut microbiota of the soil invertebrate Caenorhabditis elegans.

Plasmid conjugation contributes greatly to the spread of antibiotic resistance genes (ARGs) in soils. However, the spread potential in the gut of soil fauna remains poorly studied, and little was known about the impact of host age on ARGs dissemination in the gut microbiota of soil animals. Here, the typical nematode-Caenorhabditis elegans was employed as the model soil animal, aiming to investigate transfer of broad-host-range IncP-1ɛ from Escherichia coli MG1655 to gut microbiota within 6 days under varied temperature gradients (15, 20 and 25 °C) using qPCR combined with plate screening. Results showed that conjugation rates increased with incubation time and rising temperature in the gut of C. elegans, sharing a similar trend with abundances of plasmid conjugation relevant genes such as trbBp (mating pair formation) and trfAp (plasmid replication). Incubation time and temperature significantly shaped the gut microbial community of C. elegans. Core microbiota in the gut of C. elegans, including Enterobacteriaceae, Lactobacillaceae and Leuconostocaceae, constituted a large part of transconjugal pool for plasmid IncP-1ɛ. Our results highlight an important sink of gut microbiota for ARGs dissemination and upregulation of ARGs transfer in the gut microbiota with host age, further potentially stimulating evolution of ARGs in terrestrial environments.