Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

Objective:Based on the principle of neuroplasticity, to explore the effect of square dancing on elderly patients with subjective cognitive decline (SCD).Methods:SCD patients in the main urban area of Nanjing were selected in this quasi experimental research from March to September 2022 and divided into experimental group and control group according to whether the subjects were willing to participate in square dance intervention, with 30 cases in each group. The experimental group received 6 months of square dancing intervention, and the control group received routine nursing. The differences of cognition, memory, language, executive ability, daily living ability and anxiety and depression were compared between the two groups.Results:Twenty-seven cases in the experimental group and 28 cases in the control group completed the intervention. After intervention, the scores of Montreal Cognitive Assessment Scale-15, the Geriatric Depression Scale, Hamilton Anxiety Scale and Trail Making Test B were 27.0(26.0, 28.0), 2.0(1.0, 4.0), (4.30 ± 2.04) points and (112.48 ± 25.42) seconds, respectively. Compared with the control group of 26.0 (24.0, 27.0), 4.0 (2.0, 5.8), (5.89 ± 1.45) points and (133.21 ± 32.39) seconds, the differences were statistically significant ( Z = -2.24, -2.72, t = - 2.05, -2.64, all P<0.05). Conclusions:Square dancing can improve the cognitive function of elderly patients with SCD, promote the improvement of memory, language, executive function and daily living ability, and effectively relieve anxiety and depression symptoms.