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.

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 management system, and novel tools and methods with Chinese characteristics.
Results: The ChP 2025 edition demonstrates three prominent features in microbiological-related standards: enhanced international harmonization, introduced emerging molecular biological technologies, and established a risk-based microbiological quality control system.
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 study the effect of Liangxue Tuizi Formula(LXTZF)on reactive oxygen species(ROS)-mediated NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome activation in Henoch-Sch?nlein purpura(HSP)rats,and to explore its possible mechanism in the treatment of HSP.Methods Twenty-four rats were divided randomly into four groups:control,model,LXTZF,and compound glycyrrhizin(CG)groups.Except for the control group,a model of HSP was established in the other groups by heat drugs combined with egg albumin.After successful modeling,rats in the LXTZF group were given LXTZF solution 7.47 g/kg,rats in the CG group were given CG solution 13.5 mg/kg by gavage,and rats in the control and model groups were given normal saline solution by gavage once a day for 4 weeks.Samples were collected 8 hours after the last gavage.Skin histopathology changes were observed by hematoxylin and eosin(HE)staining.Serum interleukin(IL)-18 and IL-1βlevels were detected by enzyme-linked immunosorbent assay(ELISA).Changes in ROS levels in the skin were detected by immunofluorescence.Apoptosis-associated speckle-like protein(ASC),NLRP3,cysteinyl aspartate-specific protease-1(Caspase-1)mRNA and protein expression levels in rat skin were detected by real-time quantitative polymerase chain reaction(RT-PCR)and immunohistochemistry and Western blot,respectively.Results The skin pathology in the model group showed obvious inflammatory cell infiltration compared with the control group.Serum IL-18 and IL-1β levels were significantly increased(P＜0.05),skin ROS levels were significantly increased(P＜0.05),and skin ASC,NLRP3,Caspase-1 mRNA and protein expression levels were significantly increased(P＜0.05).Inflammatory cell infiltration in the skin tissues of rats was alleviated in the LXTZF and CG groups compared with the model group,while serum levels of IL-18 and IL-1β were significantly decreased(P＜0.05).ROS levels in the skin were significantly decreased(P＜0.05),and mRNA and protein levels of ASC,NLRP3,and Caspase-1 in the skin were significantly decreased(P＜0.05).Conclusions The mechanism of LXTZF in HSP may be related to the inhibition of ROS-mediated NLRP3 inflammasome activation.

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*

ObjectiveTo evaluate the inhibitory effect of online-sold “preservative-free” cosmetics against contaminated microorganisms during storage and use, to establish a method for assessing the preservative efficacy of such cosmetics, and to provide data support for the formulation of relevant standards. MethodsA total of 16 batches of cosmetics claiming preservative free were collected to determine their pH value, water activity, total aerobic microbial count (TAMC) and total molds and yeasts count (TYMC). Meanwhile, preservatives not listed in the Safety and Technical Standards for Cosmetics (2015 edition) (referred to as “unlisted preservatives”) were screened. In addition, a preservative challenge test was conducted on these cosmetics. ResultsAll the 16 batches of samples were generally weakly acidic, and with a water activity ≥0.6, which were suitable for microbial growth. Unlisted preservative not labeled on the package was detected in one batch of cosmetic. The results of neutralizer verification showed that three batches required further dilution to eliminate the antimicrobial interference. After inoculation with challenge microorganisms and cultivation for 7 days, two batches of cosmetics did not achieve a bacterial reduction rate of 99.90%, and the fungal reduction rate did not reach 90.00% either. While another two batches of cosmetics experienced microbial growth during testing, indicating a failure of the preservative challenge test. The overall pass rate was 75.00%. ConclusionSome online-sold preservative free cosmetics have insufficient preservation efficacy and pose a certain risk of microbial contamination.

Objective To study the regulatory effect of Qufeng Xiaodian Formula on serum differential metabolites of allergic purpura,and provide scientific basis for the diagnosis and treatment of allergic purpura in traditional Chinese medicine.Methods Sixty rats were randomly divided into a blank control group(referred to as the blank group),a model group,a compound glycyrrhizin group,a low-dose Qu Feng Xiao Dian Fang group,a medium dose Qu Feng Xiao Dian Fang group,and a high-dose Qu Feng Xiao Dian Fang group according to a random number table method,with 10 rats in each group.The model group was constructed by combining dried ginger,pepper,and long pepper with ovalbumin to create an allergic purpura rat model.After successful modeling,each treatment group was intervened with corresponding drugs for 4 weeks.After 4 weeks,serum was collected and non targeted metabolomics screening of serum differential metabolites was performed using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometer(UPLC-QTOF/MS).Subsequently,data extraction and multivariate statistical analysis will be conducted to identify potential metabolic pathways.Results Compared with the control group,there were 91 possible differential metabolites in the serum of the model group rats,corresponding to 20 metabolic pathways;Compared with the model group,there were a total of 43 possible differential metabolites in the serum of rats in the wind dispelling and disease eliminating group,corresponding to 15 metabolic pathways.Among them,there are a total of 12 metabolic pathways.Inflammatory metabolites such as arachidonic acid and ceramide can damage vascular endothelium.Ten biomarkers,including arachidonic acid and ceramide,were significantly abnormal in the serum of the model group rats compared to the normal group.The Qufeng Xiaodian formula can significantly reverse these metabolites and significantly enrich them in arachidonic acid metabolism and sphingolipid metabolism pathways.Conclusion Qufeng Xiaodian Formula has a certain regulatory effect on metabolites such as arachidonic acid and ceramide that affect vascular endothelial injury.

Objective To study the regulatory effect of Qufeng Xiaodian Formula on serum differential metabolites of allergic purpura,and provide scientific basis for the diagnosis and treatment of allergic purpura in traditional Chinese medicine.Methods Sixty rats were randomly divided into a blank control group(referred to as the blank group),a model group,a compound glycyrrhizin group,a low-dose Qu Feng Xiao Dian Fang group,a medium dose Qu Feng Xiao Dian Fang group,and a high-dose Qu Feng Xiao Dian Fang group according to a random number table method,with 10 rats in each group.The model group was constructed by combining dried ginger,pepper,and long pepper with ovalbumin to create an allergic purpura rat model.After successful modeling,each treatment group was intervened with corresponding drugs for 4 weeks.After 4 weeks,serum was collected and non targeted metabolomics screening of serum differential metabolites was performed using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometer(UPLC-QTOF/MS).Subsequently,data extraction and multivariate statistical analysis will be conducted to identify potential metabolic pathways.Results Compared with the control group,there were 91 possible differential metabolites in the serum of the model group rats,corresponding to 20 metabolic pathways;Compared with the model group,there were a total of 43 possible differential metabolites in the serum of rats in the wind dispelling and disease eliminating group,corresponding to 15 metabolic pathways.Among them,there are a total of 12 metabolic pathways.Inflammatory metabolites such as arachidonic acid and ceramide can damage vascular endothelium.Ten biomarkers,including arachidonic acid and ceramide,were significantly abnormal in the serum of the model group rats compared to the normal group.The Qufeng Xiaodian formula can significantly reverse these metabolites and significantly enrich them in arachidonic acid metabolism and sphingolipid metabolism pathways.Conclusion Qufeng Xiaodian Formula has a certain regulatory effect on metabolites such as arachidonic acid and ceramide that affect vascular endothelial injury.

Objective To study the effect of Liangxue Tuizi Formula(LXTZF)on reactive oxygen species(ROS)-mediated NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome activation in Henoch-Sch?nlein purpura(HSP)rats,and to explore its possible mechanism in the treatment of HSP.Methods Twenty-four rats were divided randomly into four groups:control,model,LXTZF,and compound glycyrrhizin(CG)groups.Except for the control group,a model of HSP was established in the other groups by heat drugs combined with egg albumin.After successful modeling,rats in the LXTZF group were given LXTZF solution 7.47 g/kg,rats in the CG group were given CG solution 13.5 mg/kg by gavage,and rats in the control and model groups were given normal saline solution by gavage once a day for 4 weeks.Samples were collected 8 hours after the last gavage.Skin histopathology changes were observed by hematoxylin and eosin(HE)staining.Serum interleukin(IL)-18 and IL-1βlevels were detected by enzyme-linked immunosorbent assay(ELISA).Changes in ROS levels in the skin were detected by immunofluorescence.Apoptosis-associated speckle-like protein(ASC),NLRP3,cysteinyl aspartate-specific protease-1(Caspase-1)mRNA and protein expression levels in rat skin were detected by real-time quantitative polymerase chain reaction(RT-PCR)and immunohistochemistry and Western blot,respectively.Results The skin pathology in the model group showed obvious inflammatory cell infiltration compared with the control group.Serum IL-18 and IL-1β levels were significantly increased(P＜0.05),skin ROS levels were significantly increased(P＜0.05),and skin ASC,NLRP3,Caspase-1 mRNA and protein expression levels were significantly increased(P＜0.05).Inflammatory cell infiltration in the skin tissues of rats was alleviated in the LXTZF and CG groups compared with the model group,while serum levels of IL-18 and IL-1β were significantly decreased(P＜0.05).ROS levels in the skin were significantly decreased(P＜0.05),and mRNA and protein levels of ASC,NLRP3,and Caspase-1 in the skin were significantly decreased(P＜0.05).Conclusions The mechanism of LXTZF in HSP may be related to the inhibition of ROS-mediated NLRP3 inflammasome activation.

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.