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 To compile the"expert consensus on preventive and management of exposure keratopathy"(hereinafter referred to as"consensus"),aiming to standardize and promote the prevention of exposure keratopathy(EK)in medical institutions at different levels.Methods The evidence-based methods were used to retrieve,evaluate and summarize evidence in this field according to the level of evidence,and relevant recommendations and research conclusions were extracted,and the first draft of consensus was formed.After 2 rounds of Delphi expert letters and expert meetings,combined with evidence and expert opinions,the consensus content is written,adjusted and modified,and recommendations were made.Results 58 experts were invited to participate in the consultation.The expert positive coefficient was 100％;the expert judgment basis of the 2 rounds were 0.940 and 0.936;the degree of familiarity of the 2 rounds were 0.779 and 0.797;the coefficient of authority of the 2 rounds were 0.859 and 0.866.The Kendall W coefficients of the 2 rounds of inquiry were 0.099 and 0.117,and the difference was statistically significant(P＜0.05).The consensus includes 5 aspects of EK:risk factors,assessments,protection measures,symptom management and training and management.Conclusion The consensus was based on the existing evidence and clinical practical requirements and recommendations of experts in the field.Consensus can provide guidance for clinical nursing staff in prevention,nursing and management of EK effectively.

Objective To investigate the distribution of traditional Chinese medicine(TCM)syndromes in chronic persistent immunoglobulin A nephropathy(IgAN)and to explore the relationship between TCM syndromes and prognosis based on real-world research.Methods From January 2018 to January 2024,a total of 80 patients with chronic persistent IgAN who were admitted to Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University were included.The clinical baseline data and prognosis-related information were collected.The main outcome indicators were defined as renal failure events of glomerular filtration rate(eGFR)decreased by＞50%,serum creatinine doubled or end-stage renal disease(ESRD).The distribution of TCM syndromes in chronic persistent IgAN and the relationship between different deficiency syndromes and prognosis were preliminarily discussed.Results(1)Among the 80 patients with chronic persistent IgAN,the syndrome manifestations with an occurrence frequency of more than 30%in descending order were foamy urine(75.00%),fatigue and lassitude(71.25%),waist ache(61.25%),dry mouth(31.25%),lower extremity edema(31.25%),aversion to cold and cold limbs(30.00%),and insomnia(30.00%).(2)The commonly-seen root-deficiency syndrome in patients with chronic persistent IgAN was qi and yin deficiency syndrome,with a total of 28 cases(35.00%).And other root-deficiency syndromes were spleen and kidney yang deficiency syndrome in 22 cases(27.50%),lung and spleen qi deficiency syndrome in 15 cases(18.75%),and liver and kidney yin deficiency syndrome in 15 cases(18.75%).The commonly-seen complicated syndrome was water-damp syndrome,with a total of 17 cases(21.25%).The other complicated syndromes were damp-heat syndrome in 15 cases(18.75%),blood-stasis syndrome in 9 cases(11.25%),phlegm-damp syndrome in 7 cases(8.75%),turbidity-toxin syndrome in 5 cases(6.25%),liver depression syndrome in 3 cases(3.75%),and cold-damp syndrome in 1 case(1.25%).(3)Among the 80 patients with chronic persistent IgAN,21 cases(26.25%)had renal failure events,and their syndrome types were lung and spleen qi deficiency syndrome in 9 cases(60.00%),qi and yin deficiency syndrome in 3 cases(10.71%),liver and kidney yin deficiency syndrome in 3 cases(20.00%),and spleen and kidney yang deficiency syndrome in 6 cases(27.27%).The incidence of renal failure events in patients with lung and spleen qi deficiency syndrome was significantly higher than that in the other three syndrome types,and the chi-square test showed that the difference was statistically significant(P＜0.05).(4)Kaplan-Meier survival analysis showed that the cumulative incidence of 2-year and 5-year endpoint events in patients with lung and spleen qi deficiency syndrome was 23.10%and 46.20%,respectively,and the final incidence was 85.00%.The cumulative incidence of 2-year and 5-year endpoint events in patients with other deficiency syndromes was 4.50%and 19.70%,respectively,and the final incidence was 71.60%.The intergroup comparison(tested by Log-rank)showed that the incidence of renal failure events in patients with lung and spleen qi deficiency syndrome was significantly higher than that in patients with other root-deficiency syndromes,and the difference was statistically significant(P＜0.05).(5)COX proportional hazard regression analysis showed that lung and spleen qi deficiency syndrome(HR=7.755,95%CI 1.631-36.874,P＜0.05)and anemia(HR=8.205,95%CI 1.007-66.857,P＜0.05)were the independent risk factors for renal failure events in patients with chronic persistent IgAN.Conclusion Qi and yin deficiency syndrome is the most common syndrome in the patients with chronic persistent IgAN,and lung and spleen qi deficiency syndrome and anemia are the independent risk factors for renal failure events in IgAN.

Objective To compile the"expert consensus on preventive and management of exposure keratopathy"(hereinafter referred to as"consensus"),aiming to standardize and promote the prevention of exposure keratopathy(EK)in medical institutions at different levels.Methods The evidence-based methods were used to retrieve,evaluate and summarize evidence in this field according to the level of evidence,and relevant recommendations and research conclusions were extracted,and the first draft of consensus was formed.After 2 rounds of Delphi expert letters and expert meetings,combined with evidence and expert opinions,the consensus content is written,adjusted and modified,and recommendations were made.Results 58 experts were invited to participate in the consultation.The expert positive coefficient was 100％;the expert judgment basis of the 2 rounds were 0.940 and 0.936;the degree of familiarity of the 2 rounds were 0.779 and 0.797;the coefficient of authority of the 2 rounds were 0.859 and 0.866.The Kendall W coefficients of the 2 rounds of inquiry were 0.099 and 0.117,and the difference was statistically significant(P＜0.05).The consensus includes 5 aspects of EK:risk factors,assessments,protection measures,symptom management and training and management.Conclusion The consensus was based on the existing evidence and clinical practical requirements and recommendations of experts in the field.Consensus can provide guidance for clinical nursing staff in prevention,nursing and management of EK effectively.

Objective To investigate the effect of COX6B2 expression in gastric cancer tissues on the patients'long-term prognosis and its underlying mechanism.Methods Based on the public databases and the medical records of patients,we analyzed the expression level of COX6B2 in gastric cancer and adjacent tissues and its influence on long-term prognosis of the patients.Enrichment analysis were used to predict the possible role of COX6B2 in gastric cancer.The effects of lentivirus-mediated COX6B2 knockdown on biological behaviors of gastric cancer cells were examined using CCK-8 assay,flow cytometry,and Western blotting.Results TCGA database and the results of immunohistochemistry,Western blotting and real-time PCR all demonstrated a significantly higher expression of COX6B2 in gastric cancer tissues(P<0.05).Kaplan-Meier plotter database and Kaplan-Meier curves showed that the patients with high COX6B2 expression had significantly shorter postoperative survival(P<0.05).A high expression of COX6B2 in gastric cancer tissues was closely correlated with clinicopathologic stage,CEA and CA19-9(P<0.05).A high expression of COX6B2,CEA level≥5 μg/L and CA19-9 level≥37 kU/L were independent risk factors affecting postoperative 5-year survival rate of gastric cancer patients(P<0.05),and COX6B2 expression level had a predictive value for long-term prognosis of the patients(P<0.05).GO and KEGG enrichment analyses showed that COX6B2 was mainly involved in the regulation of cell cycle.In the in vitro cell experiment,COX6B2 overexpression significantly promoted gastric cancer cell proliferation,increased the percentage of G1/S phase cells and inhibited the cellular expressions of p53 and p21(P<0.05).Conclusions COX6B2 is highly expressed in gastric cancer and is closely correlated with a poor long-term prognosis of the patients possibly by promoting gastric cancer cell proliferation and regulating cell cycle.

Objective To investigate the mechanism of sanguinarine(SA)for alleviating ulcerative colitis(UC)induced by dextran sodium sulfate(DSS)in mice.Methods Male C57BL/6 mouse models of 3.5%DSS-induced UC were randomized for treatment with 1,5 and 10 mg/kg SA by gavage,400 mg/kg sulfasalazine by gavage,or 10 mg/kg SA combined with intraperitoneal injection of 30 mg/kg ML385(a Nrf2 inhibitor).The changes in intestinal inflammation was assessed by monitoring weight changes,disease activity index(DAI)score,colon length measurement,and HE staining.After the treatments,the colon tissues were collected for detection of malondialdehyde(MDA)content using colorimetry,mRNA expressions of inflammatory factors using RT-qPCR,and the expressions of Nrf2,HO-1,Keap-1,p-p65,p65,occludin,and ZO-1 proteins were detected using Western blotting.Results SA treatment obviously alleviated weight loss,colon length shortening and DAI score increase and ameliorated structural destruction of the colon glands and colonic crypts in mice with DSS-induced UC.SA intervention significantly decreased the levels of TNF-α,IL-1β and IL-6 mRNA and lowered ROS and MDA levels in the colon tissue of UC mice.The mouse models receiving SA treatment showed significantly increased expressions of Nrf2,HO-1,occludin and ZO-1 and lowered expressions of Keap-1 and P-P65 in the colon tissue without significant changes of p65 expression,and these changes were SA dose-dependent.Treatment with ML385 obviously attenuated the effect of high-dose SA for improving UC in the mouse models.Conclusion SA can improve UC-like enteritis in mice possibly by activating the Nrf2 pathway and inhibiting the NF-κB pathway in the colon tissue.

To reveal the effects of N-carbamylglutamic(NCG)on growth performance,blood pa-rameters and meat quality of meat rabbits,192 Hyla rabbits at 35 days of age were assigned to four groups randomly with 0.00％,0.05％,0.10％,and 0.20％NCG added to the basal diet,with six replicates of eight rabbits in each group and one replicate of eight rabbits.The results indicated that:compared to the control group,the body weight of the 0.20％NCG group at d 35(P＜0.01),the BW at d 14 and the average daily gain(ADG)from d 1 to 14 in the 0.05％NCG group(P＜0.05)were significantly elevated;the ADG of the control group from d 1 to 35 was significantly lower than the 0.10％and 0.20％NCG groups(P＜0.05).The levels of total superoxide dismutase(T-SOD)in the 0.10％NCG group(P＜0.01),total antioxidant capacity(T-AOC)and urea nitro-gen(BUN)in the 0.20％NCG group(P＜0.05)were significantly higher compared to the control group;the levels of T-SOD in the 0.10％NCG group were significantly elevated compared to the 0.05％NCG group(P＜0.05).NCG significantly increased polyunsaturated fatty acids(PUFA)and PUFA/SFA(P＜0.05).The cooked meat rate of the longissimus lumborum in the 0.20％NCG group was significantly increased compared to the control group(P＜0.01),while the water holding rate of the longissimus lumborum increased significantly in the 0.10％NCG groups(P＜0.01)and the control group(P＜0.05)and 0.05％NCG group(P＜0.05)than the 0.20％NCG group.NCG significantly reduced the crypts depth(P＜0.01)and had the tendency to in-crease the V/C value(P=0.067),while the villi height of jejunal in the 0.20％NCG group was significantly elevated compared to the control group(P＜0.05).In conclusion,NCG could promote the growth performance,enhance the antioxidant capacity,and improve the intestinal morphology and meat quality of meat rabbits.The appropriate amount of NCG added to meat rabbit diet is 0.10％.

Objective To investigate the effect of COX6B2 expression in gastric cancer tissues on the patients'long-term prognosis and its underlying mechanism.Methods Based on the public databases and the medical records of patients,we analyzed the expression level of COX6B2 in gastric cancer and adjacent tissues and its influence on long-term prognosis of the patients.Enrichment analysis were used to predict the possible role of COX6B2 in gastric cancer.The effects of lentivirus-mediated COX6B2 knockdown on biological behaviors of gastric cancer cells were examined using CCK-8 assay,flow cytometry,and Western blotting.Results TCGA database and the results of immunohistochemistry,Western blotting and real-time PCR all demonstrated a significantly higher expression of COX6B2 in gastric cancer tissues(P<0.05).Kaplan-Meier plotter database and Kaplan-Meier curves showed that the patients with high COX6B2 expression had significantly shorter postoperative survival(P<0.05).A high expression of COX6B2 in gastric cancer tissues was closely correlated with clinicopathologic stage,CEA and CA19-9(P<0.05).A high expression of COX6B2,CEA level≥5 μg/L and CA19-9 level≥37 kU/L were independent risk factors affecting postoperative 5-year survival rate of gastric cancer patients(P<0.05),and COX6B2 expression level had a predictive value for long-term prognosis of the patients(P<0.05).GO and KEGG enrichment analyses showed that COX6B2 was mainly involved in the regulation of cell cycle.In the in vitro cell experiment,COX6B2 overexpression significantly promoted gastric cancer cell proliferation,increased the percentage of G1/S phase cells and inhibited the cellular expressions of p53 and p21(P<0.05).Conclusions COX6B2 is highly expressed in gastric cancer and is closely correlated with a poor long-term prognosis of the patients possibly by promoting gastric cancer cell proliferation and regulating cell cycle.

Objective To investigate the mechanism of sanguinarine(SA)for alleviating ulcerative colitis(UC)induced by dextran sodium sulfate(DSS)in mice.Methods Male C57BL/6 mouse models of 3.5%DSS-induced UC were randomized for treatment with 1,5 and 10 mg/kg SA by gavage,400 mg/kg sulfasalazine by gavage,or 10 mg/kg SA combined with intraperitoneal injection of 30 mg/kg ML385(a Nrf2 inhibitor).The changes in intestinal inflammation was assessed by monitoring weight changes,disease activity index(DAI)score,colon length measurement,and HE staining.After the treatments,the colon tissues were collected for detection of malondialdehyde(MDA)content using colorimetry,mRNA expressions of inflammatory factors using RT-qPCR,and the expressions of Nrf2,HO-1,Keap-1,p-p65,p65,occludin,and ZO-1 proteins were detected using Western blotting.Results SA treatment obviously alleviated weight loss,colon length shortening and DAI score increase and ameliorated structural destruction of the colon glands and colonic crypts in mice with DSS-induced UC.SA intervention significantly decreased the levels of TNF-α,IL-1β and IL-6 mRNA and lowered ROS and MDA levels in the colon tissue of UC mice.The mouse models receiving SA treatment showed significantly increased expressions of Nrf2,HO-1,occludin and ZO-1 and lowered expressions of Keap-1 and P-P65 in the colon tissue without significant changes of p65 expression,and these changes were SA dose-dependent.Treatment with ML385 obviously attenuated the effect of high-dose SA for improving UC in the mouse models.Conclusion SA can improve UC-like enteritis in mice possibly by activating the Nrf2 pathway and inhibiting the NF-κB pathway in the colon tissue.

OBJECTIVE: To analyze the expression of hydroxysteroid dehydrogenase like 2 (HSDL2) in rectal cancer tissues and the effect of changes in HSDL2 expression level on proliferation of rectal cancer cells. METHODS: Clinical data and tissue samples of 90 patients with rectal cancer admitted to our hospital from January 2020 to June 2022 were collected from the prospective clinical database and biological specimen database. The expression level of HSDL2 in rectal cancer and adjacent tissues was detected by immunohistochemistry, and based on the median level of HSDL2 expression, the patients were divided into high expression group (n=45) and low expression group (n=45) for analysis the correlation between HSDL2 expression level and the clinicopathological parameters. GO and KEGG enrichment analyses were performed to explore the role of HSDL2 in rectal cancer progression. The effects of changes in HSDL2 expression levels on rectal cancer cell proliferation, cell cycle and protein expressions were investigated in SW480 cells with lentivirus-mediated HSDL2 silencing or HSDL2 overexpression using CCK-8 assay, flow cytometry and Western blotting. RESULTS: The expressions of HSDL2 and Ki67 were significantly higher in rectal cancer tissues than in the adjacent tissues (P < 0.05). Spearman correlation analysis showed that the expression of HSDL2 protein was positively correlated with Ki67, CEA and CA19-9 expressions (P < 0.01). The rectal cancer patients with high HSDL2 expressions had significantly higher likelihood of having CEA ≥5 μg/L, CA19-9 ≥37 kU/L, T3-4 stage, and N2-3 stage than those with a low HSDL2 expression (P < 0.05). GO and KEGG analysis showed that HSDL2 was mainly enriched in DNA replication and cell cycle. In SW480 cells, HSDL2 overexpression significantly promoted cell proliferation, increased cell percentage in S phase, and enhanced the expression levels of CDK6 and cyclinD1 (P < 0.05), and HSDL2 silencing produced the opposite effects (P < 0.05). CONCLUSION The high expression of HSDL2 in rectal cancer participates in malignant progression of the tumor by promoting the proliferation and cell cycle progress of the cancer cells.
Humans ; CA-19-9 Antigen ; Ki-67 Antigen/metabolism* ; Prospective Studies ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Rectal Neoplasms/genetics* ; Cell Cycle ; Gene Expression Regulation, Neoplastic ; Hydroxysteroid Dehydrogenases/metabolism*