This study aims to investigate the effects and mechanisms of the effective-compounds of Jinshui Huanxian formula (ECC-JHF) in improving pulmonary fibrosis. Animal experiments were approved by the Ethics Committee of the Animal Experiment Center of Henan University of Chinese Medicine (approval number: IACUC-202306012). The mouse model of pulmonary fibrosis was induced using bleomycin (BLM). Hematoxylin-eosin (H&E) staining was used to detect the histopathological changes of lung tissues. Masson staining was used to assess the degree of fibrosis in lung tissues. Immunofluorescence (IF) and real-time quantitative PCR (qPCR) were performed to measure the expression of collagen type I (COL I), α-smooth muscle actin (α-SMA), fibronectin (FN), interleukin (IL)-1β, IL-6, and tumor necrosis factor α (TNF-α) in lung tissues. Flow cytometry (FCM) was employed to detect the proportion of M1 and M2 macrophages in the bronchoalveolar lavage fluid (BALF) of mice. IF and qPCR were also used to detect the expression of lipase family member N (LIPN) in lung tissues. Free fatty acid assay kit was used to detect the level of free fatty acids in lung tissue. Bone marrow-derived macrophages (BMDMs) were treated with interleukin-4 (IL-4) to induce M2 polarization. FCM was used to measure the proportion of CD206⁺ M2 macrophages. IF was utilized to detect LIPN expression and lipid droplet decomposition. The results showed that in BLM-induced pulmonary fibrosis mice, ECC-JHF significantly attenuated BLM-induced alveolar inflammation and collagen deposition, inhibited fibroblast activation in lung tissues, and decreased the proportion of M2 macrophages in BALF. It also significantly suppressed LIPN expression and free fatty acid level in lung tissues. In the IL-4 induced BMDMs M2 polarization model, ECC-JHF significantly inhibited the proportion of CD206⁺ M2 macrophages, down-regulated the expression of LIPN, and blocked lipid droplet catabolism. These results suggest that ECC-JHF may alleviate bleomycin-induced pulmonary fibrosis by inhibiting lipid droplet decomposition and M2 macrophage polarization.

ObjectiveTo investigate the current status of latent tuberculosis infection (LTBI) among the elderly population in rural areas of Changxing County, Zhejiang Province, and to provide an evidence for the development of LTBI prevention and control measures. MethodsBetween January and May 2024, elderly individuals participating in urban and rural residents’ health checkups were screened for Mycobacterium tuberculosis infection using a domestically produced interferon-γ release assay (IGRA) kit. Individuals tested positive by IGRA but without active tuberculosis were classified as LTBI cases. The prevalence of LTBI among the participants was subsequently analyzed. ResultsAmong the 6 765 subjects, 637 tested positive by IGRA, including one identified active tuberculosis patient, resulting in a LTBI prevalence rate of 9.40%. There was a statistically significant difference in positivity rates across different IGRA methodologies (χ²=35.530, P<0.001). Higher LTBI rate was observed in males, individuals with a history of diabetes mellitus, and those with a history of pulmonary tuberculosis, exhibiting statistically significant differences (χ²=32.401, P<0.001; χ²=5.789, P=0.020; χ²=39.248, P<0.001, respectively.) No statistically significant difference in LTBI rate was found across different age groups (χ²=0.238, P=0.971). ConclusionThe prevalence of LTBI among the elderly rural residents in Changxing County is relatively low. Male, individuals with a history of diabetes mellitus, and those with a history of pulmonary tuberculosis have an increased risk of LTBI, warranting targeted risk monitoring and timely interventions.

Conventional chemotherapy based on cytotoxic drugs is facing tough challenges recently following the advances of monoclonal antibodies and molecularly targeted drugs. It is critical to inspire new potential to remodel the value of this classical therapeutic strategy. Here, we fabricate bisphosphonate coordination lipid nanogranules (BC-LNPs) and load paclitaxel (PTX) to boost the chemo- and immuno-therapeutic synergism of cytotoxic drugs. Alendronate in BC-LNPs@PTX, a bisphosphonate to block mevalonate metabolism, works as both the structure and drug constituent in nanogranules, where alendronate coordinated with calcium ions to form the particle core. The synergy of alendronate enhances the efficacy of paclitaxel, suppresses tumor metastasis, and alters the cytotoxic mechanism. Differing from the paclitaxel-induced apoptosis, the involvement of alendronate inhibits the mevalonate metabolism, changes the mitochondrial morphology, disturbs the redox homeostasis, and causes the accumulation of mitochondrial ROS and lethal lipid peroxides (LPO). These factors finally trigger the ferroptosis of tumor cells, an immunogenic cell death mode, which remodels the suppressive tumor immune microenvironment and synergizes with immunotherapy. Therefore, by switching paclitaxel-induced apoptosis to mevalonate metabolism-triggered ferroptosis, BC-LNPs@PTX provides new insight into the development of cytotoxic drugs and highlights the potential of metabolism regulation in cancer therapy.

Endo-beta-N-acetylglucosaminidase (ENGase) is widely distributed in various organisms. The first reported ENGase activity was detected in Diplococcus pneumoniae in 1971. The protein (Endo D) was purified and its peptide sequence was determined in 1974. Three ENGases (Endo F1-F3) were discovered in Flavobacterium meningosepticum from 1982 to 1993. After that, the activity was detected from different species of bacteria, yeast, fungal, plant, mice, human, etc. Multiple ENGases were detected in some species, such as Arabidopsis thaliana and Trichoderma atroviride. The first preliminary crystallographic analysis of ENGase was conducted in 1994. But to date, only a few ENGases structures have been obtained, and the structure of human ENGase is still missing. The currently identified ENGases were distributed in the GH18 or GH85 families in Carbohydrate-Active enZyme (CAZy) database. GH18 ENGase only has hydrolytic activity, but GH85 ENGase has both hydrolytic and transglycosylation activity. Although ENGases of the two families have similar (β/α)8-TIM barrel structures, the active sites are slightly different. ENGase is an effective tool for glycan detection andglycan editing. Biochemically, ENGase can specifically hydrolyze β‑1,4 glycosidic bond between the twoN-acetylglucosamines (GlcNAc) on core pentasaccharide presented on glycopeptides and/or glycoproteins. Different ENGases may have different substrate specificity. The hydrolysis products are oligosaccharide chains and a GlcNAc or glycopeptides or glycoproteins with a GlcNAc. Conditionally, it can use the two products to produce a new glycopeptides or glycoprotein. Although ENGase is a common presentation in cell, its biological function remains unclear. Accumulated evidences demonstrated that ENGase is a none essential gene for living and a key regulator for differentiation. No ENGase gene was detected in the genomes of Saccharomyces cerevisiae and three other yeast species. Its expression was extremely low in lung. As glycoproteins are not produced by prokaryotic cells, a role for nutrition and/or microbial-host interaction was predicted for bacterium produced enzymes. In the embryonic lethality phenotype of the Ngly1-deficient mice can be partially rescued by Engase knockout, suggesting down regulation of Engase might be a solution for stress induced adaptation. Potential impacts of ENGase regulation on health and disease were presented. Rabeprazole, a drug used for stomach pain as a proton inhibitor, was identified as an inhibitor for ENGase. ENGases have been applied in vitro to produce antibodies with a designated glycan. The two step reactions were achieved by a pair of ENGase dominated for hydrolysis of substrate glycoprotein and synthesis of new glycoprotein with a free glycan of designed structure, respectively. In addition, ENGase was also been used in cell surface glycan editing. New application scenarios and new detection methods for glycobiological engineering are quickly opened up by the two functions of ENGase, especially in antibody remodeling and antibody drug conjugates. The discovery, distribution, structure property, enzymatic characteristics and recent researches in topical model organisms of ENGase were reviewed in this paper. Possible biological functions and mechanisms of ENGase, including differentiation, digestion of glycoproteins for nutrition and stress responding were hypothesised. In addition, the role of ENGase in glycan editing and synthetic biology was discussed. We hope this paper may provide insights for ENGase research and lay a solid foundation for applied and translational glycomics.

Protein as the allergens could lead to allergy. In addition, a widespread class of allergens were known as glycans of N-glycoprotein. N-glycoprotein contained oligosaccharide linked by covalent bonds with protein. Recently,studies implicated that allergy was associated with glycans of heterologous N-glycoprotein found in food, inhalants, insect toxins, etc. The N-glycan structure of N-glycoprotein allergen has exerted an influence on the binding between allergens and IgE, while the recognition and presentation of allergens by antigen-presenting cells (APCs) were also affected. Some researches showed thatN-glycan structure of allergen was remodeled by N-glycosidase, such as cFase I, gpcXylase, as binding of allergen and IgE partly decreased. Thus, allergic problems caused by N-glycoproteins could potentially be solved by modifying or altering the structure ofN-glycoprotein allergens, addressing the root of the issue. Mechanism of N-glycans associated allergy could also be elaborated through glycosylation enzymes, alterations of host glycosylation. This article hopes to provide a separate insight for glycoimmunology perspective, and an alternative strategy for clinical prevention or therapy of allergic diseases.

ObjectiveHuman Ku70 protein mainly involves the non-homologous end joining (NHEJ) repair of double-stranded DNA breaks (DSB) through its DNA-binding properties, and it is recently reported having an RNA-binding ability. This paper is to explore whether Ku70 has RNA helicase activity and affects miRNA maturation. MethodsRNAs bound to Ku protein were analyzed by RNA immunoprecipitation sequencing (RIP-seq) and bioinfomatic anaylsis. The expression relationship between Ku protein and miRNAs was verified by Western blot (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays. Binding ability of Ku protein to the RNAs was tested by biolayer interferometry (BLI) assay. RNA helicase activity of Ku protein was identified with EMSA assay. The effect of Ku70 regulated miR-124 on neuronal differentiation was performed by morphology analysis, WB and immunofluorescence assays with or without Zika virus (ZIKV) infection. ResultsWe revealed that the Ku70 protein had RNA helicase activity and affected miRNA maturation. Deficiency of Ku70 led to the up-regulation of a large number of mature miRNAs, especially neuronal specific miRNAs like miR-124. The knockdown of Ku70 promoted neuronal differentiation in human neural progenitor cells (hNPCs) and SH-SY5Y cells by boosting miR-124 maturation. Importantly, ZIKV infection reduced the expression of Ku70 whereas increased expression of miR-124 in hNPCs, and led to morphologically neuronal differentiation. ConclusionOur study revealed a novel function of Ku70 as an RNA helicase and regulating miRNA maturation. The reduced expression of Ku70 with ZIKV infection increased the expression of miR-124 and led to the premature differentiation of embryonic neural progenitor cells, which might be one of the causes of microcephaly.

OBJECTIVE To compare the changes of chemical components of Morus alba leaves， screen differential markers， and determine their contents， so as to provide reference for quality control of M. alba leaves before and after baked with honey. METHODS The fingerprints of M. alba leaves before and after baked with honey were established by high-performance liquid chromatography （HPLC）. The common peaks of the fingerprints were identified and the similarity was evaluated. The differential markers of M. alba leaves before and after baked with honey were screened by principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） using common peak are of raw material and product baked with honey of M. alba leaves as index. The quantitative analysis was carried out. RESULTS Twenty-three and twenty-four common peaks were identified from the HPLC fingerprint spectra of ten batches of raw material and ten batches of product baked with honey of M. alba leaves， respectively. The similarities of HPLC fingerprints for raw material and product baked with honey of M. alba leaves were all greater than 0.97. The results of PCA showed that raw material and product baked with honey of M. alba leaves could be divided into two categories. The results of OPLS-DA showed that the variable importance in projection of peak 2， peak H （5- hydroxymethylfurfural）， peak 1， peak 17 （isochlorogenic acid C） and peak 16 were all greater than 1. The average contents of differential marker of isochlorogenic acid C in raw material and product baked with honey of M. alba leaves were 0.093 6 and 0.127 8 mg/g， respectively； there was statistical significance （P＜0.05）. CONCLUSIONS Five differential markers such as isochlorogenic acid C are obtained. The content of isochlorogenic acid C in M. alba leaves is significantly increased after baked with honey.

Cryptogenic multifocal ulcerous stenosing enteritis（CMUSE） is a rare and difficult disease characte-rized by chronic recurrent intestinal obstruction and gastrointestinal bleeding caused by multiple ulcers and strictures of the small intestine. There is still a lack of mature and systematic guidance for the treatment of the disease. This paper reported a case of CMUSE mainly manifested as fatigue， abdominal distention， and edema of both lower limbs ， who is treated by integrated traditional Chinese and western medicine. In western medicine treatment， hormone and symptomatic treatment are mainly used. In traditional Chinese medicine treatment， the method of fortifying spleen and reinforcing healthy qi is taken as the basis； during the active stage of the disease， the method of moving qi and removing stagnation， clearing and removing dampness and heat， astringing and engendering flesh should be combined with， while during the remission stage， it is advised to unite with the method of diffusing lung and moving water， regulating and harmonizing zang-fu （脏腑） organs； simultaneously， replenishing qi and invigorating blood can be used throughout the treatment， and attach importance to the maintenance therapy of the traditional Chinese medicine immunomo-dulatory agent Tripterygium glycosides. After more than 2 years of treatment， the patient's clinical symptoms were significantly relieved， and all indicators basically restored to normal， having a stable condition.

Stress urinary incontinence (SUI) and underactive bladder (UAB) are common types of lower urinary tract dysfunction in women.As the treatment mechanisms of the two conditions are contradictory, the treatment of SUI patients complicated with UAB remains a difficult clinical problem.In order to improve the treatment rate of such patients and promote research, this paper reviews the latest domestic and overseas diagnostic criteria of UAB, summarizes the treatment experience of conventional midurethral sling (tension-free vaginal tape or outside-in transobturator tape) and adjustable sling procedures (transobturator adjustable tape or Remeex system) combined with medication or intermittent catheterization, and the application prospects of cutting-edge technologies such as stem cell injection, cytokine therapy and gene therapy, so as to provide reference for clinicians and researchers.

Objective To explore the correlation between speech fluency and diadochokinetic rate in children aged 7～11 years with cochlear implant.Methods Speech samples were collected from 62 children aged 7～11 years with cochlear implant using language retelling task and diadochokinetic rate test task.Their speech rate,articulation rate,syllable duration and pause duration were analyzed.The data were input into ICF converter to obtain the im-pairment limit of each parameter and analyze the speech fluency and diadochokinetic rate characteristics.Partial cor-relation analysis was performed for speech rate,articulation rate,syllable duration and pause duration by two tasks.Results ① For children with cochlear implant,the mean ICF impairment limit of diadochokinetic rate was 1.3± 0.1,the mean ICF impairment limit of fluency in retelling was 1.0±0.3,with various degree of impairment in both tasks.② There was a moderate or low correlation between speech rate,articulation rate,syllable duration in retell-ing task and those in diadochokinetic rate task(0.3＜|r |≤0.8).There was a low correlation between pause dura-tion in retelling task and speech rate and pause duration in diadochokinetic rate task(0.3＜|r |≤0.5).Conclusion The speech fluency and diadochokinetic rate of children aged 7～11 years with cochlear implant is underdeveloped,and the poor articulation movement ability limits their development of speech fluency.