Objective To evaluate the safety of the 13-valent pneumococcal polysaccharide conjugate vaccine (tetanus toxoid/diphtheria toxoid) (PCV13-TT/DT) among age-eligible children in Tianjin through a combination of active and passive surveillance methods. Methods From July 15, 2023, to August 31, 2024, active surveillance for adverse events following immunization (AEFI) was conducted among recipients of PCV13-TT/DT at 18 selected vaccination clinics in Tianjin. Recipients were monitored through on-site observation or telephone follow-up within 30 minutes after vaccination and on days 1, 3, 7, and 28. Passive surveillance for AEFI was conducted among recipients of PCV13-TT/DT at other vaccination clinics across the city. The incidence of AEFI was analyzed using descriptive epidemiological methods. Results A total of 24 916 recipients of PCV13-TT/DT were observed, with 440 AEFI cases reported, resulting in an overall incidence rate of 176.59 per 10 000. The incidence rate of AEFI in active surveillance was 813.79 per 10 000, significantly higher than that in passive surveillance (20.49 per 10 000; P< 0.001). The incidence rates of general reactions, abnormal reactions, and coincidental cases in active surveillance were 744.44 per 10 000, 8.16 per 10 000, and 61.19 per 10 000, respectively, all of which were higher than those in passive surveillance (18.49 per 10 000, 0.50 per 10 000, and 1.50 per 10 000), with P values < 0.05. General reactions were mainly characterized by fever, local redness, and local induration. Abnormal reactions included angioedema and allergic rash. Coincidental cases were mainly infections. No severe adverse reactions occurred. Conclusion The large-scale vaccination of PCV13-TT/DT after its launch has good safety, and continuous strengthening of vaccine safety monitoring is needed.

Objective: To observe the effect of Xipayimaizibizi oral liquid (XP) in an overactive bladder (OAB) experimental rat model and to explore its pharmacological mechanisms. Methods: Network pharmacology was used to explore the potential mechanisms of action of XP. The rats underwent bladder outlet obstruction surgery and were administered the corresponding drug concentrations by gavage for 4 weeks. The study observed the body weight, water intake, bladder and kidney indices (to evaluate their general status), urination behavior pattern (to observe frequency and urgency), and urodynamics (to measure bladder parameters). Hematoxylin and eosin and Masson's trichome staining were used to observe changes in the bladder structure. Enzyme-linked immunosorbent assay was used to measure the levels of nerve growth factor, brain-derived neurotrophic factor, and acetylcholine in the urine. The key targets involved in these mechanisms were validated using reverse transcription-quantitative polymerase chain reaction, immunohistochemistry, and western blot in vivo/vitro experiments. Result: Network pharmacological analysis predicted that XP may alleviate OAB by affecting the cholinergic synapse and calcium signaling pathways. XP treatment significantly reduced the bladder index, improved urine behavior and urodynamic parameters, decreased the neurotransmitters in urine, and reduced the thickness of the bladder wall and collagen ratio. These results indicate that XP can alleviate OAB symptoms and improve the bladder structure. In vivo/vitro experiments further demonstrated that XP can inhibit targets, such as muscarinic acetylcholine receptor 2, and participate in cholinergic synapses to further regulate the parasympathetic nervous system. It can also reduce the overexpression of Ca2+ caused by agonists, inhibit targets such as transient receptor potential vanilloid type 1, and participate in calcium signaling pathways to maintain Ca2+ homeostasis. Conclusion These results suggest that XP inhibited bladder overactivity by maintaining Ca2+ homeostasis and regulating the parasympathetic nervous system.

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 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 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 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.

As people's living standards improve， the development trend of diabetes has gradually become severe. Diabetes is a chronic inflammatory disease associated with abnormal expression of nuclear factor-kappa B （NF-κB） in patients. NF-κB exists in various tissue cells and participates in the regulation of a variety of genes related to immune function and inflammation. Varieties of factors can activate NF-κB when the body is stimulated by external factors， so as to produce inflammation and other reactions. Previous studies on NF-κB mainly focus on cancer， and the pathological mechanism of the treatment of diabetes by related signaling pathways and the progress of traditional Chinese medicine （TCM） treatment have not been systematically elaborated on. By referring to the relevant literature in China and abroad， it was found that NF-κB is not isolated in the development and progression of diabetes but is associated with signal molecules related to inflammation， oxidative stress， and energy metabolism， and it is involved in mediating inflammation， pancreatic β cell apoptosis， insulin signal transduction， and other physiological functions. Therefore， blocking the transmission of NF-κB signaling pathway is beneficial to the treatment of diabetes. At present， Western medicine for the treatment of diabetes mainly includes oral hypoglycemic drugs and insulin injections， but the adverse reactions are obvious. TCM has been characterized by multi-target， extensive action， and excellent curative effects in the treatment of diabetes. TCM and its compounds with functions of tonifying Qi and promoting blood circulation， regulating qi and eliminating phlegm， clearing heat and detoxifying， and nourishing Yin and moistening dryness can effectively intervene in the abnormal expression of NF-κB signaling pathway in vivo through anti-inflammatory effects. In this paper， the association between NF-κB signaling pathway and diabetes was summarized， and the modern research progress of TCM intervention of NF-κB signaling pathway in the treatment of diabetes in the past five years was reviewed， so as to lay a laboratory foundation for the study of a new pathological mechanism of diabetes based on NF-κB signaling pathway and provide new targets and research direction for the prevention and treatment of diabetes and development of related TCM.

ObjectiveTo observe the effect of water extract of Mori Folium （MLE） on oxidative stress in adipose tissue of type 2 diabetes mellitus （T2DM） mice and explore its mechanism. MethodTwenty-four male db/db mice were randomly divided into model group， metformin group， low-dose MLE （MLE-L） group， and high-dose MLE （MLE-H） group according to their body weight and blood glucose， with six mice in each group， and other six C57BLKS/JGpt wild littermate mice were selected as normal group. The mice in the metformin group were given 200 mg·kg^-1 metformin suspension， and the mice in the MLE-L and MLE-H groups were respectively given 2 g·kg^-1 and 4 g·kg^-1 MLE， while the mice in the normal group and model group were given the same dose of deionized water by daily gavage for eight weeks. Body weight， subcutaneous fat index， fasting blood glucose （FBG）， and oral glucose tolerance level （OGTT） of the mice were detected， and serum superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and malondialdehyde （MDA） were measured. The expression levels of silent information regulator 1 （SIRT1） and NADPH oxidase type 4 （NOX4） protein in subcutaneous adipose tissue of the mice were detected by Western blot. ResultThe FBG level， OGTT， and subcutaneous fat index of T2DM mice were significantly decreased （P<0.05， P<0.01） after administration of MLE compared with the blank group. The contents of serum SOD and GSH were significantly increased， while the level of oxidative stress damage marker MDA was significantly decreased （P<0.05， P<0.01）. The expression of SIRT1 protein in adipose tissue was significantly increased， while the expression of NOX4 protein was significantly decreased （P<0.05， P<0.01）. ConclusionMLE can ameliorate T2DM by alleviating oxidative stress in adipose tissue of T2DM mice and reducing blood glucose.