OBJECTIVES: To investigate the subcellular localization and biological functions of transmembrane protein 240 (TMEM240). METHODS: NCBI BLAST and TMHMM bioinformatics software were used for protein sequence analysis and prediction of transmembrane domain of TMEM240. Brain tissues from male C57BL/6 mice (18-20 days old) were examined for distribution of TMEM240 using in situ hybridization, and qPCR and Western blotting were used to detect TMEM240 expression in different mouse tissues and in cortical neurons at different time points (n=3). In the in vitro experiment, HepG2 and Neuro-2a cells were transfected with plasmids for overexpression of TMEM240, and subcellular localization of TMEM240 was analyzed using cell imaging. In primary cultures of cortical neurons isolated from C57BL/6 mice, TMEM240 expression and its biological functions were investigated using qPCR, Western blotting, and immunofluorescence staining. RESULTS: Human and mouse TMEM240 proteins share a 97.69% similarity in the protein sequences, and both are transmembrane proteins with two transmembrane domains. TMEM240 mRNA and protein were highly expressed in mouse brain tissues and cortical neurons. In isolated mouse cortical neurons, TMEM240 expression reached the peak level after primary culture for 9 days and distributed in scattered spots within the cells. In HepG2 cells, TMEM240 was characterized as intracellular membrane structures and showed 80% colocalization with peroxisomes. In Neuro-2a cells, TMEM240 overexpression caused significant enhancement of the expressions of Rho GDP dissociation inhibitor β (ARHGDIB) at both the mRNA and protein levels. CONCLUSIONS TMEM240 is a novel intracellular subcellular structure specifically expressed in neurons with significant potential for targeted cellular function regulation.
Animals ; Humans ; Mice ; Peroxisomes/metabolism* ; Membrane Proteins/genetics* ; Mice, Inbred C57BL ; Neurons/metabolism* ; Male ; rho-Specific Guanine Nucleotide Dissociation Inhibitors ; Hep G2 Cells ; Brain/metabolism*

Objective:To investigate the differences in resistance to chemical disinfectants between bacteriophage Phi-X174 and disinfection effect indicator microorganisms, Staphylococcus aureus and Escherichia coli. Methods:Three commonly used disinfectants, including chlorine-containing disinfectants, alcohol-based disinfectants and quaternary ammonium salt disinfectants, were selected to analyze the differences in resistance of phage Phi-X174, Staphylococcus aureus and Escherichia coli by using the quantitative sterilization test of suspension. Results:The 250 mg/L sodium dichloroisocyanurate solution treated for 10 minutes yielded log reduction values of 3.39, 7.25 and 7.37 against phage Phi-X174, Staphylococcus aureus and Escherichia coli respectively. The 40% ethanol for 1 minute yielded log reduction values of 0.03, 2.46 and 7.30 against phage Phi-X174, Staphylococcus aureus and Escherichia coli, respectively. The 100 mg/L benzalkonium chloride for 10 minutes yielded log reduction values of 1.41, 6.84 and 0.93 for phage Phi-X174, Staphylococcus aureus and Escherichia coli, respectively. Conclusion:Phage Phi-X174 has stronger resistance to chlorine-containing disinfectants and alcohol-based disinfectants than Escherichia coli and Staphylococcus aureus. Its resistance to quaternary ammonium salt disinfectants is between that of Staphylococcus aureus and Escherichia coli.

Objective:To systematically analyze the revisions content and technological development trends of microbiological standards in the Chinese Pharmacopoeia(ChP)2025 Edition,and explore its novel requirements in risk-based pharmaceutical product lifecycle management.Methods:A comprehensive review was conducted on 26 microbiological-related standards to summarize the revision directions and scientific implications from perspectives including the revision overview,international harmonization of microbiological standards,risk-based quality man-agement system,and novel tools and methods with Chinese characteristics.Results:The ChP 2025 edition demon-strates three prominent features in microbiological-related standards:enhanced international harmonization,intro-duced emerging molecular biological technologies,and established a risk-based microbiological quality control sys-tem.Conclusion:The new edition of the Pharmacopoeia has systematically constructed a microbiological standard system,which significantly improves the scientificity,standardization and applicability of the standards,providing a crucial support for advancing the microbiological quality control in pharmaceutical industries of China.

The microbial limit standards in the Chinese Pharmacopoeia 2020 edition primarily focus on total aerobic microbial count and specified objectionable microorganisms,which are insufficient for comprehensively assessing the potential risks posed by microbial contamination to drug efficacy and patient safety.With the increasing com-plexity of rising regulatory requirements,there is an urgent need to establish a scientific and systematic microbial risk assessment and control framework.In response,the Chinese Pharmacopoeia 2025 edition,introduces a new general guideline 9212 Risk Assessment and Control of Unacceptable Microorganisms for Non-sterile Products.This chapter systematically constructs a framework for the identification and control of risks associated with unacceptable microorganisms,filling a gap in the relevant field within international pharmacopoeias.This article systematically elaborates on its key elements based on its core framework and content.It covers the development background,core concepts,testing and identification strategies,evaluation of risk characterization factors,formulation of risk control measures,and implementation pathways.The aim is to provide pharmaceutical enterprises and regulatory agencies with systematic and clear practical guidance.

Objective:To investigate the differences in resistance to chemical disinfectants between bacteriophage Phi-X174 and disinfection effect indicator microorganisms, Staphylococcus aureus and Escherichia coli. Methods:Three commonly used disinfectants, including chlorine-containing disinfectants, alcohol-based disinfectants and quaternary ammonium salt disinfectants, were selected to analyze the differences in resistance of phage Phi-X174, Staphylococcus aureus and Escherichia coli by using the quantitative sterilization test of suspension. Results:The 250 mg/L sodium dichloroisocyanurate solution treated for 10 minutes yielded log reduction values of 3.39, 7.25 and 7.37 against phage Phi-X174, Staphylococcus aureus and Escherichia coli respectively. The 40% ethanol for 1 minute yielded log reduction values of 0.03, 2.46 and 7.30 against phage Phi-X174, Staphylococcus aureus and Escherichia coli, respectively. The 100 mg/L benzalkonium chloride for 10 minutes yielded log reduction values of 1.41, 6.84 and 0.93 for phage Phi-X174, Staphylococcus aureus and Escherichia coli, respectively. Conclusion:Phage Phi-X174 has stronger resistance to chlorine-containing disinfectants and alcohol-based disinfectants than Escherichia coli and Staphylococcus aureus. Its resistance to quaternary ammonium salt disinfectants is between that of Staphylococcus aureus and Escherichia coli.

The microbial limit standards in the Chinese Pharmacopoeia 2020 edition primarily focus on total aerobic microbial count and specified objectionable microorganisms,which are insufficient for comprehensively assessing the potential risks posed by microbial contamination to drug efficacy and patient safety.With the increasing com-plexity of rising regulatory requirements,there is an urgent need to establish a scientific and systematic microbial risk assessment and control framework.In response,the Chinese Pharmacopoeia 2025 edition,introduces a new general guideline 9212 Risk Assessment and Control of Unacceptable Microorganisms for Non-sterile Products.This chapter systematically constructs a framework for the identification and control of risks associated with unacceptable microorganisms,filling a gap in the relevant field within international pharmacopoeias.This article systematically elaborates on its key elements based on its core framework and content.It covers the development background,core concepts,testing and identification strategies,evaluation of risk characterization factors,formulation of risk control measures,and implementation pathways.The aim is to provide pharmaceutical enterprises and regulatory agencies with systematic and clear practical guidance.

The immune related adverse events(irAE)caused by tumor immune checkpoint inhibitors(ICI)have attracted increasing attention of clinical experts.Immune-mediated liver injury caused by ICIs(ILICI)is not uncommon in clinical practice,but specific diagnostic method of ILICI is lacking.Biopsy of liver tissue can help improve the diagnosis and management of ILICI.In the treatment of ILICI,the immediate use of corticosteroid therapy is not necessarily.A balance between efficacy,toxicity,and specific treatment need to be achieved,and further refined through multidisciplinary team(MDT)cooperation.Appropriate dosaging and identification of novel predictive targets should be considered in order to reduce the incidence and severity of ILICI in the future.Meanwhile,further basic research is required to elucidate the potential pathophysiological mechanisms and risk factors of ILICI.With the refinement of evidence in clinical evidence-based medicine and deepening of basic research,the diagnosis and treatment level of ILICI will also be further improved.

The main pathogenesis of chronic cough in children is the disorder of ascending and descending of qi movement caused by healthy qi deficiency and pathogenic qi retention. The deficiency of lung， spleen， and kidney is the root of the disease， and the retention of phlegm-fluid， food accumulation， and fire from constraint is the branch pathogenesis of the disease. In the treatment， we should reinforce and tonify healthy qi， dispel pathogen and regulate qi， with Yupingfeng Powder （玉屏风散） as the basic prescription. For lung qi deficiency syndrome， modified Yupingfeng Powder could be used for supplementing lung to consolidate the exterior； for lung and spleen qi deficiency syndrome， modified Yupingfeng Powder plus Shenling Baizhu Powder （参苓白术散） could be used for supplementing lung and fortifying the spleen， treating with both supplementation and transformation； for lung kidney qi deficiency syndrome， modified Yupingfeng Powder combined with Suzi Jiangqi Decoction （苏子降气汤） could be used for supplementing lung and replenishing kidneys， absorbing qi to the root. All the above prescriptions could combine the method of dispelling phlegm， promoting digestion and guiding out food stagnation， soothing the liver and draining fire to remove the solid pathogens， in order to treat the root and branch simultaneously， and the cough will stop if the ascending and descending of qi movement recover as usual.

ObjectiveTo observe the effect of Coptidis Rhizoma and Ophiopogonis Radix on renal tissue injury and epidermal growth factor receptor （EGFR）/phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway in rats with diabetic nephropathy （DN） and explore its possible mechanism of delaying DN. MethodThirty-six male Wistar rats were randomly divided into a normal group （6 rats） and a model group （30 rats）. The model group was fed with a high-fat and high-sugar diet combined with streptozotocin （STZ） to establish a rat model of type 2 diabetes. After the successful preparation of the model， the rats were randomly divided into the model group， low， medium， and high dose groups of Coptidis Rhizoma and Ophiopogonis Radix （100， 200， 400 mg·kg^-1）， and metformin group （200 mg·kg^-1）. After administration， the levels of fasting blood glucose （FBG）， 24 h urine protein （24 h-UTP）， creatinine （SCr）， urea nitrogen （BUN）， and uric acid （UA） were detected. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the pathological changes of renal tissue in rats. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to detect the related protein expression of EGFR， PI3K， and Akt and their mRNA expression levels in the renal tissue of rats in each group. ResultsCompared with the normal group， the levels of FBG， SCr， BUN， UA， 24 h-UTP， and kidney index in the model group were significantly increased （P<0.01）， most renal tubular epithelial cells were necrotic， and the content of collagen in glomeruli was significantly increased （P<0.01）. Compared with the model group， the above indexes of rats in each administration group were improved to varying degrees. The FBG， SCr， BUN， UA， 24 h-UTP， and kidney index of rats in each dose group and metformin group were significantly decreased （P<0.01， P<0.05）. The necrosis degree of renal tubular epithelial cells was reduced， and the fibrosis area was decreased （P<0.01）. There related protein and mRNA expressions of EGFR， PI3K， and Akt were significantly increased （P<0.05， P<0.01）. ConclusionCoptidis Rhizoma and Ophiopogonis Radix can alleviate renal tissue injury in rats with DN， and their mechanism may be related to the regulation of the EGFR/PI3K/Akt signaling pathway.

Asthma is a common chronic respiratory disease characterized by repeated attacks and prolonged illness.Traditional Chinese medicine believes that the formation of Sugen is the core pathogenesis of repeated asthma attacks.By tracing the origin of Sug-en theory,summarizing the connotation of ancient asthma Sugen theory and the innovative understanding of modern medical scholars on asthma Sugen,this paper explores the potential connection between the traditional Chinese medicine Sugen theory and the pathogenesis of modern asthma,in order to provide new ideas and methods for the treatment and research of asthma.