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*

Fracture healing is a complex process that necessitates the synergistic action of various cells and molecules. Macrophages play an indispensable and crucial regulatory role in the process of fracture repair, influencing stages such as inflammatory modulation, angiogenesis, and tissue remodeling. This article delves into the functional characteristics of macrophages and their roles at different stages of fracture healing. Additionally, it explores the impact of aging macrophages on the healing process. Furthermore, the potential of emerging therapies, such as hydrogel-based treatments and exosomes, in modulating macrophage responses is analyzed. This study provides a theoretical foundation for the development of innovative therapies aimed at enhancing the efficacy of fracture healing.

Objective:Metabolic alkalosis has raised concerns in patients receiving continuous renal replacement therapy (CRRT) via regional citrate anticoagulation (RCA). This study searched for alkalosis-related factors and mechanisms.Methods:It's a retrospective study of alkalosis in patients who received CRRT for at least 12 hours with RCA in a tertiary hospital between April 2017 and April 2020. Demographic features, baseline laboratory results, CRRT metrics and laboratory results at 12h after CRRT was recorded. Patients was grouped based on whether alkalosis exist at 12h after CRRT, and multivariable logistic regression analysis was used to identify risk factors for alkalosis during CRRT with citrate anticoagulation.Results:The 59 patients meeting the inclusion criteria were 49% male, with a mean age of (55±18) years old, and 42% had alkalosis by 12 hours after CRRT. No significant differences in demographic features or laboratory results were observed patients with or without alkalosis. CRRT metrics, including blood flow rate, citrate rate, replacement fluid rate and total effluent rate, were significantly different among groups ( P<0.01). Multivariable Logistic regression analysis indicated that the citrate rate was a risk factor for alkalosis ( OR=1.088, 95% CI 1.020-1.161, P =0.010). In patients receiving no NaHCO 3 and without alkalosis, the linear regression analysis described the relationships of citrate with replacement fluid rate (citrate rate = 0.090 × replacement fluid rate + 56.581; R2 = 0.6918) and total effluent rate (citrate rate = 0.099 × total effluent rate + 2.449). Conclusions:This retrospective observational study demonstrated that CRRT metrics are highly associated with alkalosis after 12 hours of CRRT. Without NaHCO3 infusion, a 10-fold linear correlation was observed between citrate and total effluent rate in patients without metabolic alkalosis.

Objective To analyze the risk factors for the development of chronic critical illness(CCI)in septic shock patients in the intensive care unit(ICU).Methods A retrospective analysis was conducted on 102 sepsis shock patients admitted to the ICU of the hospital from August 2018 to May 2023.According to their clinical dates,they were divided into CCI group(n=60)and rapid recovery(RR)group(n=42).Multi-ple logistic regression analysis was perform to identify independent risk factors for developing CCI in septic shock patients,and construct their receiver operating characteristic(ROC)curves to evaluate their predictive value for CCI.Results The prolonged duration of hypernatremia(OR=1.770,95％CI:1.147-2.731,P=0.010)and low acute physiology and chronic health(APACHEⅡ)scores(OR=1.212,95％CI:1.025-1.433,P=0.025)were independent risk factors for CCI in ICU septic shock patients.The occurrence of acute gastrointestinal injury(OR=0.060,95％CI:0.011-0.341,P=0.002)and peak blood sodium levels during ICU(OR=0.812,95％CI:0.662-0.995,P=0.044)were protective factors.Conclusion For septic shock pa-tients,special attention should be paid to its related independent risk factors and protective factors to improve the prognosis of septic shock patients.

Objective:To identify the serotype and genotype characteristics of the first cluster infection of Brucella in Nujiang Lisu Autonomous Prefecture of Yunnan Province. Methods:The information of seven suspected cluster brucellosis cases reported in Lanping County, Nujiang Lisu Autonomous Prefecture in April 2023 were collected, blood samples were collected, and the strains were isolated and cultured. Serological methods were employed for brucellosis diagnosis, bacterial species identification was performed using BCSP31-PCR and AMOS-PCR. Multilocus locus variable-number tandem repeat analysis (MLVA) was conducted for strain genotyping, followed by phylogenetic analysis comparing the strains with those from other regions of China.Results:All seven cases were diagnosed with brucellosis, they were all villagers from the same village and had daily contact with sheeps. Four suspected Brucella strains were isolated, identified as Brucella melitensis biotype by BCSP31-PCR and AMOS-PCR. The four isolated strains exhibited identical MLVA-16 genotype, with MLVA-8 genotype 42 and MLVA-11 genotype 180, belonging to the Eastern Mediterranean lineage. Compared with the MLVA-16 of the isolated strains from other regions of China, the four isolated strains formed an independent cluster. Conclusion:The first cluster infection of Brucella in Nujiang Lisu Autonomous Prefecture of Yunnan Province is caused by Brucella melitensis biotype, and the four isolated strains have showed unique MLVA genotype.

In July 2024,the Diagnosis Related Groups(DRG)2.0 is released based on the Notice from the National Healthcare Security Administration on Issuing the DRG 2.0 and Deepening the Relevant Work.Compared with DRG 1.1,version 2.0 was established based on a wider range of suggestions regarding the Adjacent Diagnosis Related Groups(ADRG),Major Comorbidity or Complication(MCC),and Comorbidity or Complication(CC)from various institutions.A list of disease diagnoses and surgical operations that are not used as grouping rules was compiled,and grouping efficacy was further improved by upgrading the algorithms for MCC and CC with the help of AI.Meanwhile,it is necessary to pay more attention to the number of cases of ADRG,the better methods to list the MCC/CC,the suggestions of various doctors and continuously standardize the data and update the grouping scheme of DRG.

In July 2024,the Diagnosis Related Groups(DRG)2.0 is released based on the Notice from the National Healthcare Security Administration on Issuing the DRG 2.0 and Deepening the Relevant Work.Compared with DRG 1.1,version 2.0 was established based on a wider range of suggestions regarding the Adjacent Diagnosis Related Groups(ADRG),Major Comorbidity or Complication(MCC),and Comorbidity or Complication(CC)from various institutions.A list of disease diagnoses and surgical operations that are not used as grouping rules was compiled,and grouping efficacy was further improved by upgrading the algorithms for MCC and CC with the help of AI.Meanwhile,it is necessary to pay more attention to the number of cases of ADRG,the better methods to list the MCC/CC,the suggestions of various doctors and continuously standardize the data and update the grouping scheme of DRG.

Objective:To identify the serotype and genotype characteristics of the first cluster infection of Brucella in Nujiang Lisu Autonomous Prefecture of Yunnan Province. Methods:The information of seven suspected cluster brucellosis cases reported in Lanping County, Nujiang Lisu Autonomous Prefecture in April 2023 were collected, blood samples were collected, and the strains were isolated and cultured. Serological methods were employed for brucellosis diagnosis, bacterial species identification was performed using BCSP31-PCR and AMOS-PCR. Multilocus locus variable-number tandem repeat analysis (MLVA) was conducted for strain genotyping, followed by phylogenetic analysis comparing the strains with those from other regions of China.Results:All seven cases were diagnosed with brucellosis, they were all villagers from the same village and had daily contact with sheeps. Four suspected Brucella strains were isolated, identified as Brucella melitensis biotype by BCSP31-PCR and AMOS-PCR. The four isolated strains exhibited identical MLVA-16 genotype, with MLVA-8 genotype 42 and MLVA-11 genotype 180, belonging to the Eastern Mediterranean lineage. Compared with the MLVA-16 of the isolated strains from other regions of China, the four isolated strains formed an independent cluster. Conclusion:The first cluster infection of Brucella in Nujiang Lisu Autonomous Prefecture of Yunnan Province is caused by Brucella melitensis biotype, and the four isolated strains have showed unique MLVA genotype.

OBJECTIVE To establish a comprehensive platform for regional pharmaceutical care among Jiangxi provincial pediatric alliance to realize the management of pediatric hierarchical diagnosis and treatment, and improve the quality of pharmaceutical care. METHODS A unified diagnosis and treatment information standard and a knowledge base of children's rational drug use rules were established among the medical institutions of Jiangxi provincial pediatric alliance. On this basis, the medical records and drug use information of patients in various medical institutions in the region were uploaded to the chain in a structured manner in real time, and a comprehensive platform for regional pharmaceutical care was built. RESULTS The comprehensive platform for regional pharmaceutical care built based on blockchain technology could share medical resources and information among medical institutions, realize rational drug use management, remote prescription review, individualized drug use guidance, popular science education, government supervision, etc., and improve the quality of pharmaceutical care. CONCLUSION The comprehensive platform for regional pharmaceutical care among Jiangxi provincial pediatric alliance can help allocate high-quality medical resources (drug safety knowledge base and pediatric pharmacists) for primary medical institutions. Further more, it lays the foundation for government supervision at the same time ensuring children’s medication safety, which has great practical significance.

Objective:To study the effects of poly adenosine diphosphate ribose polymerase (PARP) inhibitors niraparib and pamiparib on the radiosensitivity of breast cancer cell lines MCF-7 and MDA-MB-436, and to explore its mechanism.Methods:MCF-7 and MDA-MB-436 cells were divided into control group, niraparib group, pamiparib group, radiation group, combination group treated with niraparib and radiation, and combination group treated with pamiparib and radiation, respectively. The effects of drugs on cell proliferation and radiosensitivity were measured by CCK-8 assay and colony formation assay, respectively. The effect of drugs combined with radiation on cell cycle and apoptosis were detected by flow cytometry. Immunofluorescence method was used to detect the changes of γ-H2AX focal number of cells. The expressions of FANCG, Bax and Bcl-2 mRNA and protein were detected by qPCR and Western blot, respectively.Results:Both niraparib and pamiparib inhibited the proliferation of breast cancer cells MCF-7 and MDA-MB-436 in a time-dose dependent manner. With the increase of irradiation dose, D0, Dq, SF2 value of MCF-7 and MDA-MB-436 cells decreased, and SER D0 and SER Dq value increased. Compared with control group, the percentages of cells in G 2/M phase were increased ( tMCF-7=41.66, 44.08, P<0.05; t436=24.69, 18.91, P<0.05), the percentage of cells in G 0/G 1 phase were decreased ( tMCF-7=8.67, 29.61, P<0.05; t436=26.39, 29.12, P<0.05), and the cell apoptosis rate was significantly increased ( tMCF-7=11.17, 11.71, P<0.05; t436=42.68, 15.89, P<0.05) in the combination group. Compared with control group, the number of γ-H2AX foci of MCF-7 cells in the radiation group and combination group treated with niraparib and radiation increased significantly at 2 h after irradiation ( t=8.89, 21.72, P<0.05). At 24 h after irradiation, the number of γ-H2AX foci basically returned to normal level in the radiation group but remained at a higher level in the combination group ( t=8.82, P<0.05). Compared with control group, the expressions of FANCG and Bcl-2 mRNA decreased ( tFANCG=14.07, P<0.05; tBcl-2=29.21, P<0.05), the expression of Bax mRNA increased ( t=8.90, P<0.05), and the expression of FANCG and Bcl-2 proteins decreased ( tFANCG=7.09, P<0.05; tBcl-2=10.24, P<0.05), while the expression of Bax protein increased ( t=2.90, P<0.05) in the combination group. Conclusions:PARP inhibitors niraparib and pamiparib can increase the radiosensitivity of breast cancer MCF-7 and MDA-MB-436 cells probably through down-regulating the expression of FANCG in FA-BRCA pathway, up-regulating apoptosis-related genes and inhibiting DNA damage repair.