[Intestinal absorption components and absorption characteristics of Wuwei Qingzhuo San by everted intestinal sac models].

This study investigated the absorption profile of Wuwei Qingzhuo San in different intestinal segments and the absorption characteristics of its alkaloids(piperine, piperanine, piperlonguminine, and dihydropiperlonguminine). The everted gut sac model was established, and the chemical components of Wuwei Qingzhuo San in different intestinal segments were detected by UPLC-Q-TOF-MS. The content of piperine, piperanine, piperlonguminine, and dihydropiperlonguminine in intestinal absorption fluid was determined by UPLC-Q-TRAP-MS and the absorption parameters were calculated. The absorption characteristics in different intestinal segments at different time were analyzed. As a result, 27, 27, 8, and 6 absorbent components from Wuwei Qingzhuo San were detected in the intestinal cyst fluid of jejunum, ileum, duodenum, and colon by UPLC-Q-TOF-MS technology, respectively. It was also found that piperine, piperanine, piperlonguminine, and dihydropiperlonguminine from Wuwei Qingzhuo San showed linear absorption in various intestinal segments, with r values exceeding 0.9. In terms of absorption content, the components were ranked as piperine>piperanine>dihydropiperlonguminine>piperlonguminine in various intestinal segments, but the absorption rate and mechanism of each component varied. The results demonstrate that the absorption of the components of Wuwei Qingzhuo San in different intestinal segments is selective and is not a simple semi-permeable membrane permeation process.

Antibiotic resistance bacteria and antibiotic resistance genes survived from the extremely acidity posing a risk on intestinal bacteria in an in vitro digestion model by horizontal gene transfer.

Antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) are emerging contaminants posing risk to human health. To investigate the pathogenic ARBs and the horizontal gene transfer (HGT) via both extracellular ARGs (eARGs) and intracellular ARGs (iARGs), an in vitro digestion simulation system was established to monitoring the ARB and ARGs passing through the artificial digestive tract. The results showed that ARB was mostly affected by the acidity of the gastric fluid with about 99% ARB (total population of 2.45 × 109-2.54 × 109) killed at pH 2.0 and severe damage of bacterial cell membrane. However, more than 80% ARB (total population of 2.71 × 109-3.90 × 109) survived the challenge when the pH of the gastric fluid was 3.0 and above. Most ARB died from the high acidity, but its ARGs, intI1 and 16 S rRNA could be detected. The eARGs (accounting for 0.03-24.56% of total genes) were less than iARGs obviously. The eARGs showed greater HGT potential than that of iARGs, suggesting that transformation occurred more easily than conjugation. The transferring potential followed: tet (100%) > sul (75%) > bla (58%), related to the high correlation of intI1 with tetA and sul2 (p < 0.01). Moreover, gastric juice of pH 1.0 could decrease the transfer frequency of ARGs by 2-3 order of magnitude compared to the control, but still posing potential risks to human health. Under the treatment of digestive fluid, ARGs showed high gene horizontal transfer potential, suggesting that food-borne ARBs pose a great risk of horizontal transfer of ARGs to intestinal bacteria.

[Textual research principles and suggestions on function and indication of ancient classical formulas].

Indication and functions is an important key information in the research and development of the ancient classical formulas, which directly affects the clinical positioning of the compound formulas and their reasonable, effective and safe use after marke-ting. It is also the embodiment of the ultimate vitality of ancient classical formulas. Due to the particularity of ancient classical formulas, it is of great significance to accurately define and describe the functions and indications of classical formulas to exert their unqiue advantages. Based on the analysis of the provenances, classification of clinical indications, and functions of 100 prescriptions in the Catalogue of Ancient Classical Formulas(First Batch), this paper summarized the incompleteness, irregularity and inconsistency in the original text, the differences in terminology between ancient and modern times, and the evolution of the meanings of prescriptions in different dynasties. In addition, under the guidance of the general principle of textual researh on ancient classical formulas, which is to inherit the essence, make the past serve the present, link up the past and the present, and bulid consensus, this paper proposed the following ideas: respecting the original meanings and provenances of ancient classical formulas, taking full consideration of the development and evolution, serving the current clinical application and health needs, accurately linking up the ancient and modern terminologies, standardizing the expression of traditional Chinese medicine(TCM) terminology, highlighting the characteristics of TCM, attaching importance to the textual research principles and suggestions of post-marketing evidence-based and clinical positioning research, so as to determine the the functions and indications of the ancient classical formulas in a scientific, standardized and reasonable way and better guide the clinical application of the classical formulas.

Simultaneous reduction of antibiotics and antibiotic resistance genes in pig manure using a composting process with a novel microbial agent.

In recent years, in order to promote animal growth and reduce the risk of disease, a variety of antibiotics are frequently added to the animal feed of livestock and poultry. However, these antibiotics can not be fully digested by animals and most of them are excreted with feces, consequently causing the enrichment of antibiotic resistance genes (ARGs) and huge environmental risks. Nowadays, composting is a better option to solve these problems. Accordingly, this study explored the effects of co-composting swine manure with different inoculants dominated by Phanerochaete chrysosporium (p), Aspergillus niger (a), and Bacillus licheniformis (b) on the simultaneous removal of multiple antibiotics and resistance genes and evolution of the bacterial community. The results showed that the highest removal extent of tetracycline and oxytetracycline occurred in pile D (p:b:a=1:5:5, biomass) reaching 89.2% and 87.8%, respectively, while the highest removal extent of doxycycline and enrofloxacin occurred in pile A (p:b:a=1:0:0, biomass) reaching 98.6% and 89%, respectively. Compared with the levels in pile B (control check), in pile D, ARGs, except those for sulfonamides, decreased by 1.059 × 10-3-6.68 × 10-2 gene copies/16S rRNA copies. Inoculation with p alone effectively reduced intI1 and intI2. Canonical correspondence analysis (CCA) that microbial community structure evolution had a greater influence on ARGs than environmental factors. In summary, this study provided a feasible way to efficiently remove the antibiotics and antibiotic resistance genes in pig manure.

Spread of resistance genes from duck manure to fish intestine in simulated fish-duck pond and the promotion of cefotaxime and As.

Integrated culture is a widespread culture mode in South China, in which resistance genes (RGs) also spread in the circulation system with nutrients. Accordingly, the aim of the present study was to investigate the spread of RGs in a fish-duck pond and the RGs and bacterial community of fish intestines. Five fish tanks, including a control tank and four experimental tanks (duck manure, duck manure + cefotaxime, duck manure + As, and duck manure + cefotaxime + As), were tested for 100 days. The results showed that duck manure increased both the diversity and relative abundance of RGs in fish intestines, and the addition of stress factors (cefotaxime, As) increased the relative abundance of RGs by one to two orders of magnitude. The stress-inducing effect of cefotaxime was greater than that of As. Tetracycline resistance genes were more sensitive to stress factors and were the predominant RGs in fish intestines. RGs in duck manure preferentially spread from the water to biofilm and then to fish intestines, whereas co-stress of cefotaxime and As obviously promoted the spread of RGs to fish intestines. In comparison to the control tank, duck manure and stress factors significantly changed the bacterial community of fish intestines. Correlation analysis also revealed that arsB, MOX, tetA and sul1 were significantly correlated with intI1 (P < 0.01), which hinted a potentially dissemination risk of RGs in fish intestines. These findings provide a theoretical basis for further investigating the dissemination of RGs in integrated culture systems and for evaluating the ecological risk of antibiotic and As use in aquaculture.

Evolution and distribution of resistance genes and bacterial community in water and biofilm of a simulated fish-duck integrated pond with stress.

Integrated fish-duck pond is a common circular farming model in South China, besides, it is also hot-spot for the co-selection of antibiotic resistance genes (ARGs). The aim of this study was to investigate the effects of duck manure, As and cefotaxime on the bacterial community, and the evolution and distribution of ARGs and metal(loid) resistance genes (MRGs) in water and biofilm. Five groups of fish tanks included a control and four test groups. The experimental period lasted for 100 days. Six ARGs (CIT, DHA, EBC, FOX, MOX, TEM), two MRGs (arsB, arsC), and two integron genes (intI1, intI2) were tracked and detected in water and biofilm. The results showed that duck manure brought ARGs and MRGs into fish tanks. Stress factors (cefotaxime, As) increased the relative abundance of resistance genes, and this was positively correlated with stress concentrations. The biofilm was visible significantly at the end of stage 3, and the total relative abundance of resistance genes in biofilm was higher than water from stage 5 onwards. Evolution of AmpC ß-lactamase resistance genes was more obvious than MRGs, especially for MOX, which increased by 3 orders of magnitude. The abundance of Flavobacterium was higher in biofilm than in water. Moreover, correlation analysis showed that both arsB and MOX were significantly correlated with intI1 (p < 0.05), which suggested a potentially dissemination risk of resistance genes. This study provides a reference for health risk assessment in integrated aquaculture environment contaminated with duck manure, antibiotics and metalloids.