Objective To develop a core outcome set(COS)for clinical effectiveness trials of heart failure with preserved ejection fraction(HFpEF).Methods Outcome measures were collected through database literatures search,clinical experts questionnaire survey and semi-structured patients interview.Then,the outcome measures pool was constructed and domains were divided.Candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,a consensus meeting was held to determine COS and reach a consensus.Results A total of 317 outcome measures which could be divided into 6 domains were collected through literature research,questionnaire survey and semi-structured interview.15 candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,the consensus meeting reached consensus on a COS with 6 entries.Conclusion In this study,a COS for clinical effectiveness trials of HFpEF was developed,which is conducive to the standardization of efficacy evaluation.

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 explore the expression of 7-dehydrocholesterol reductase (DHCR7) in gastric cancer using bioinformatics methods and its relationship with clinical pathological characteristics and prognosis of gastric cancer patients.Methods:DHCR7 expression in gastric cancer was analyzed using the UALCAN database; DHCR7 mRNA expression and its relationship with the prognosis of gastric cancer patients were analyzed using the Kaplan-Meier plotter database; The expression of DHCR7 and its correlation with tumor immune infiltration level were analyzed using Sangerbox 3.0 and TIMER database; Real-time fluorescence quantitative PCR was used to detect the expression of DHCR7 mRNA in gastric cancer tissues and adjacent tissues; immunohistochemical staining was conducted to detect the DHCR7 expression in gastric cancer tissues and adjacent tissues and its correlation with clinical pathological parameters; Receiver operator characteristic (ROC) curve was used to evaluate the efficacy of DHCR7 expression in the diagnosis of gastric cancer.Results:The analysis results of the UALCAN database showed that there were statistically significant differences in DHCR7 mRNA expression among gastric cancer patients of different genders ( χ2=18.15, P<0.001), grades ( χ2=16.32, P<0.001), and TP53 mutation status ( χ2=20.12, P<0.001). Survival analysis showed that the 10-year overall survival (OS) rate ( HR=1.55, 95% CI: 1.31-1.84, P<0.001), 10-year progression free survival (PFS) rate ( HR=1.67, 95% CI: 1.36-2.05, P<0.001), and 10-year post progression survival (PPS) rate ( HR=1.81, 95% CI: 1.43-2.28, P<0.001) of gastric cancer patients with high DHCR7 expression were significantly lower than those with low DHCR7 expression. Immune infiltration analysis showed the expression of DHCR7 was negatively correlated with the comprehensive score ( r=-0.51, P<0.001), stromal cell score ( r=-0.48, P<0.001), immune cell score ( r=-0.45, P<0.001), CD4 + T cells ( r=-3.01, P<0.001), macrophages ( r=-0.40, P<0.001), neutrophils ( r=-0.32, P<0.001), and dendritic cells ( r=-0.37, P<0.001) infiltration levels in gastric cancer, and positively correlated with the purity of gastric cancer cells ( r=0.15, P<0.001). The qRT-PCR results showed that compared with adjacent tissues (1.86±0.51), the expression of DHCR7 in gastric cancer tissues (3.43±0.13) was significantly upregulated, with a statistically significant difference ( t=42.89, P<0.001). The relative expression level of DHCR7 in normal gastric mucosal cells GES-1 was 1.06±0.19, and the relative expression levels in four types of gastric cancer cells (HGC-27, AGS, SNU-1, and SGC-7901) were 2.40±0.26, 1.88±0.11, 1.51±0.04, and 2.63±0.20, respectively, there were statistically significant differences in the expression of DHCR7 among the five types of cells ( F=38.34, P<0.001), and the relative expression level of DHCR7 in normal gastric mucosal cells was statistically significant different compared to the four types of gastric cancer cells mentioned above ( P=0.002; P=0.003; P=0.017; P<0.001) ; The immunohistochemical results showed that the high expression rate of DHCR7 in gastric cancer tissues was 80.0% (96/120), which was significantly higher than that in adjacent tissues (68.3%) (82/120) ( χ2=56.84, P<0.001). There were statistically significant differences in tumor maximum diameter ( χ2=40.17, P<0.001), histological grade ( χ2=16.20, P<0.001) and pTNM stage ( χ2=16.99, P<0.001) between patients with high and low DHCR7 expression. The ROC curve results showed that the area under the curve (AUC) of DHCR7 expression level for diagnosing gastric cancer were 0.76 (based on TCGA database, 95% CI: 0.68-0.83, P<0.001) and 0.97 (120 clinical samples of gastric cancer, 95% CI: 0.95-0.99, P<0.001), respectively. Conclusions:DHCR7 is highly expressed in gastric cancer and closely associated with poor prognosis in patients, which may be a novel biomarker for the diagnosis and prognosis of gastric cancer.

Objective To investigate the efficacy and safety of Linggui Qihua No.2 Prescription(LGQH2)in patients with heart failure with preserved ejection fraction(HFpEF).Methods Totally 60 HFpEF patients were randomly divided into experimental group and control group according to random number table method,with 30 patients in each group.On the basis of standardized treatment for heart failure,the experimental group was given LGQH2 granules,13 g/time,twice a day,orally;the control group was given placebo granules of LGQH2,with the same administration method as the experimental group.The treatment course for both groups was 4 weeks.6-minute walking distance(6MWD),Kansas City Cardiomyopathy Questionnaire(KCCQ)score,TCM syndrome score and serum NT-proBNP levels.Echocardiography was used to detect the ratio of early diastolic blood flow velocity(E)at the mitral valve to early diastolic myocardial motion velocity(e')at the mitral annulus,left atrial diameter(LAD),left ventricular end-diastolic diameter(LVEDD)and interventricular septal thickness(IVST).Adverse reactions and events were also recorded.Results Compared with before treatment,both groups showed significant improvement in 6MWD,KCCQ score,TCM syndrome score,serum NT proBNP,and E/e'after treatment(P<0.05),and there was no statistical significance in LAD,LVEDD and IVST between the two groups(P>0.05);after treatment,the experimental group showed better improvement in 6MWD,KCCQ score,TCM syndrome score and E/e'compared to the control group(P<0.05).During the research process,neither group of patients experienced any adverse reactions or events.Conclusion LGQH2 Prescription can effectively enhance exercise tolerance and cardiac function of HFpEF patients,alleviate symptoms,improve quality of life,and inhibit diastolic dysfunction of the heart,without notable adverse reactions.

Objective To explore the mechanism of Scutellaria baicalensis(Huangqin)in ameliorating postoperative cognitive dysfunction in surgical patients through network pharmacology approaches combined with experimental validation.Methods Network pharmacology was used to screen the relevant targets and pathways of Huangqin in relation to postoperative cognitive dysfunction,followed by molecular docking to verify the affinity of core components with key targets.HT22 cell line of mouse hippocampal neurons was cultured,and an inflammation injury model was induced by LPS stimulation.Huangqin was selected for intervention treatment,divided into five groups:control group,model group,low-dose Huangqin group(10 μmol/L),medium-dose Huangqin group(50 μmol/L),and high-dose Huangqin group(100 μmol/L).After co-culturing for 24 h,the expression of key target proteins HIF1A,MAPK1(ERK2/p-ERK2),and SIRT1 were detected by Western blotting,and the mRNA expression of these proteins was measured by qRT-PCR.Results A total of 27 active components of Huangqin were screened by network pharmacology,corresponding to 379 targets,with 220 disease targets related to POCD,resulting in 35 intersecting genes.Molecular docking results showed that components like baicalein and oroxylin A had strong affinity with targets such as HIF1A,MAPK1,and SIRT1.In vitro experimental results indicated that baicalein significantly downregulated the expression and transcription levels of HIF1A and MAPK1(ERK2/p-ERK2),while upregulating the expression and transcription levels of SIRT1,effectively improved the inflammatory response in HT22 cells and reduced neuronal damage.Conclusion The traditional Chinese medicine Huangqin can reduce the occurrence and development of postoperative cognitive dysfunction by improving the expression of key proteins,controlling inflammatory responses,and protecting neuronal function.

BACKGROUND:The role of microRNA-214 in osteoporosis has been reported both at home and abroad,whereas the interrelationship between microRNA-214 and osteoarthritic articular cartilage and subchondral bone degeneration is unclear. OBJECTIVE:To investigate the relationship between microRNA-214 and cartilage and subchondral bone degeneration in mice with knee osteoarthritis. METHODS:Thirty C57BL/6J mice were randomly grouped:Experiment 1:sham operation group and medial meniscus destabilization group (n=3 per group) underwent hematoxylin-eosin staining and qPCR to detect changes in microRNA-214 gene expression;Experiment 2:sham operation group,medial meniscus destabilization group,medial meniscus destabilization+null-loaded adenovirus group (null-loaded group),and medial meniscus destabilization+microRNA-214 antagonist overexpression adenovirus group (antagonist group;n=6 per group). Cartilage tissues were taken from each group 4 weeks after surgery,and stained with hematoxylin-eosin,safranin O-fast green,and toluidine blue. qPCR and western blot were used to detect the expression of related factors in articular cartilage. RESULTS AND CONCLUSION:(1) In Experiment 1,hematoxylin-eosin staining results showed that cartilage degeneration was visible in the medial meniscus destabilization group compared with the sham operation group. qPCR assay results showed that microRNA-214 was expressed in all the samples,and the expression level of microRNA-214 in cartilage samples of the medial meniscus destabilization group was significantly higher than that of the sham operation group (P<0.05). (2) In Experiment 2,the results of hematoxylin-eosin staining,safranin O-fast green staining,and toluidine blue staining showed that the degree of cartilage degeneration in the antagonist group was significantly reduced compared with the medial meniscus destabilization group. Adenovirus-validated PCR assay showed that the microRNA-214 expression level in cartilage tissue was higher in the null-loaded group than in the antagonist group (P<0.05). (3) In Experiment 2,X-ray results showed typical osteoarthritis imaging changes in the medial meniscus destabilization group and null-loaded group,while the degree of degenerative joint lesions was relatively mild in the antagonist group. The results of microcomputed tomography showed that after injection of microRNA-214 antagonist,trabecular structure model index became smaller in the antagonist group,and the data were better than those of the medial meniscus destabilization group and null-loaded group. (4) In Experiment 2,western blot results showed that The relative expression levels of cartilage-associated factor type Ⅱ collagen α1,sex-determining region Y-box 9,Runt-associated transcription factor 2,and osteopontin in cartilage specimens of the medial meniscus destabilization group and the null-loaded group were lower than that in the sham operation group and the antagonist group (P<0.05),whereas the relative expression level of matrix metalloproteinase 13 was higher in the medial meniscus destabilization group and the null-loaded group than the sham operation group and the antagonist group (P<0.05). (5) In Experiment 2,PCR results indicated that the relative mRNA expression of tumor necrosis factor-α and interleukin-6 was relatively higher in the medial meniscus destabilization group and null-loaded group,but relatively lower in the antagonist group,as compared with the sham operation group (P<0.05). The relative mRNA expression of tumor necrosis factor-α and interleukin-6 was also higher in the medial meniscus destabilization group and the null-loaded group compared with the antagonist group (P<0.05). To conclude,the expression level of microRNA-214 in articular cartilage was elevated in the mouse osteoarthritis model,suggesting that the elevated expression level of microRNA-214 is closely related to osteoarthritis;and injection of microRNA-214 antagonist into the knee joint cavity of the mouse osteoarthritis model could delay articular cartilage degradation,promote subchondral bone remodeling,and ameliorate the progression of osteoarthritis.

Objective To develop a core outcome set(COS)for clinical effectiveness trials of heart failure with preserved ejection fraction(HFpEF).Methods Outcome measures were collected through database literatures search,clinical experts questionnaire survey and semi-structured patients interview.Then,the outcome measures pool was constructed and domains were divided.Candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,a consensus meeting was held to determine COS and reach a consensus.Results A total of 317 outcome measures which could be divided into 6 domains were collected through literature research,questionnaire survey and semi-structured interview.15 candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,the consensus meeting reached consensus on a COS with 6 entries.Conclusion In this study,a COS for clinical effectiveness trials of HFpEF was developed,which is conducive to the standardization of efficacy evaluation.

Objective To explore the mechanism of Scutellaria baicalensis(Huangqin)in ameliorating postoperative cognitive dysfunction in surgical patients through network pharmacology approaches combined with experimental validation.Methods Network pharmacology was used to screen the relevant targets and pathways of Huangqin in relation to postoperative cognitive dysfunction,followed by molecular docking to verify the affinity of core components with key targets.HT22 cell line of mouse hippocampal neurons was cultured,and an inflammation injury model was induced by LPS stimulation.Huangqin was selected for intervention treatment,divided into five groups:control group,model group,low-dose Huangqin group(10 μmol/L),medium-dose Huangqin group(50 μmol/L),and high-dose Huangqin group(100 μmol/L).After co-culturing for 24 h,the expression of key target proteins HIF1A,MAPK1(ERK2/p-ERK2),and SIRT1 were detected by Western blotting,and the mRNA expression of these proteins was measured by qRT-PCR.Results A total of 27 active components of Huangqin were screened by network pharmacology,corresponding to 379 targets,with 220 disease targets related to POCD,resulting in 35 intersecting genes.Molecular docking results showed that components like baicalein and oroxylin A had strong affinity with targets such as HIF1A,MAPK1,and SIRT1.In vitro experimental results indicated that baicalein significantly downregulated the expression and transcription levels of HIF1A and MAPK1(ERK2/p-ERK2),while upregulating the expression and transcription levels of SIRT1,effectively improved the inflammatory response in HT22 cells and reduced neuronal damage.Conclusion The traditional Chinese medicine Huangqin can reduce the occurrence and development of postoperative cognitive dysfunction by improving the expression of key proteins,controlling inflammatory responses,and protecting neuronal function.

Objective To investigate the efficacy and safety of Linggui Qihua No.2 Prescription(LGQH2)in patients with heart failure with preserved ejection fraction(HFpEF).Methods Totally 60 HFpEF patients were randomly divided into experimental group and control group according to random number table method,with 30 patients in each group.On the basis of standardized treatment for heart failure,the experimental group was given LGQH2 granules,13 g/time,twice a day,orally;the control group was given placebo granules of LGQH2,with the same administration method as the experimental group.The treatment course for both groups was 4 weeks.6-minute walking distance(6MWD),Kansas City Cardiomyopathy Questionnaire(KCCQ)score,TCM syndrome score and serum NT-proBNP levels.Echocardiography was used to detect the ratio of early diastolic blood flow velocity(E)at the mitral valve to early diastolic myocardial motion velocity(e')at the mitral annulus,left atrial diameter(LAD),left ventricular end-diastolic diameter(LVEDD)and interventricular septal thickness(IVST).Adverse reactions and events were also recorded.Results Compared with before treatment,both groups showed significant improvement in 6MWD,KCCQ score,TCM syndrome score,serum NT proBNP,and E/e'after treatment(P<0.05),and there was no statistical significance in LAD,LVEDD and IVST between the two groups(P>0.05);after treatment,the experimental group showed better improvement in 6MWD,KCCQ score,TCM syndrome score and E/e'compared to the control group(P<0.05).During the research process,neither group of patients experienced any adverse reactions or events.Conclusion LGQH2 Prescription can effectively enhance exercise tolerance and cardiac function of HFpEF patients,alleviate symptoms,improve quality of life,and inhibit diastolic dysfunction of the heart,without notable adverse reactions.

BACKGROUND:The role of microRNA-214 in osteoporosis has been reported both at home and abroad,whereas the interrelationship between microRNA-214 and osteoarthritic articular cartilage and subchondral bone degeneration is unclear. OBJECTIVE:To investigate the relationship between microRNA-214 and cartilage and subchondral bone degeneration in mice with knee osteoarthritis. METHODS:Thirty C57BL/6J mice were randomly grouped:Experiment 1:sham operation group and medial meniscus destabilization group (n=3 per group) underwent hematoxylin-eosin staining and qPCR to detect changes in microRNA-214 gene expression;Experiment 2:sham operation group,medial meniscus destabilization group,medial meniscus destabilization+null-loaded adenovirus group (null-loaded group),and medial meniscus destabilization+microRNA-214 antagonist overexpression adenovirus group (antagonist group;n=6 per group). Cartilage tissues were taken from each group 4 weeks after surgery,and stained with hematoxylin-eosin,safranin O-fast green,and toluidine blue. qPCR and western blot were used to detect the expression of related factors in articular cartilage. RESULTS AND CONCLUSION:(1) In Experiment 1,hematoxylin-eosin staining results showed that cartilage degeneration was visible in the medial meniscus destabilization group compared with the sham operation group. qPCR assay results showed that microRNA-214 was expressed in all the samples,and the expression level of microRNA-214 in cartilage samples of the medial meniscus destabilization group was significantly higher than that of the sham operation group (P<0.05). (2) In Experiment 2,the results of hematoxylin-eosin staining,safranin O-fast green staining,and toluidine blue staining showed that the degree of cartilage degeneration in the antagonist group was significantly reduced compared with the medial meniscus destabilization group. Adenovirus-validated PCR assay showed that the microRNA-214 expression level in cartilage tissue was higher in the null-loaded group than in the antagonist group (P<0.05). (3) In Experiment 2,X-ray results showed typical osteoarthritis imaging changes in the medial meniscus destabilization group and null-loaded group,while the degree of degenerative joint lesions was relatively mild in the antagonist group. The results of microcomputed tomography showed that after injection of microRNA-214 antagonist,trabecular structure model index became smaller in the antagonist group,and the data were better than those of the medial meniscus destabilization group and null-loaded group. (4) In Experiment 2,western blot results showed that The relative expression levels of cartilage-associated factor type Ⅱ collagen α1,sex-determining region Y-box 9,Runt-associated transcription factor 2,and osteopontin in cartilage specimens of the medial meniscus destabilization group and the null-loaded group were lower than that in the sham operation group and the antagonist group (P<0.05),whereas the relative expression level of matrix metalloproteinase 13 was higher in the medial meniscus destabilization group and the null-loaded group than the sham operation group and the antagonist group (P<0.05). (5) In Experiment 2,PCR results indicated that the relative mRNA expression of tumor necrosis factor-α and interleukin-6 was relatively higher in the medial meniscus destabilization group and null-loaded group,but relatively lower in the antagonist group,as compared with the sham operation group (P<0.05). The relative mRNA expression of tumor necrosis factor-α and interleukin-6 was also higher in the medial meniscus destabilization group and the null-loaded group compared with the antagonist group (P<0.05). To conclude,the expression level of microRNA-214 in articular cartilage was elevated in the mouse osteoarthritis model,suggesting that the elevated expression level of microRNA-214 is closely related to osteoarthritis;and injection of microRNA-214 antagonist into the knee joint cavity of the mouse osteoarthritis model could delay articular cartilage degradation,promote subchondral bone remodeling,and ameliorate the progression of osteoarthritis.