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.

Objective To enhance the understanding of the quality control of rubber closures and to promote pharmaceutical manufacturers'reasonable selection and effective management of rubber closures.Methods According to the performance needs of drug products for rubber closures,the impacts of the ingredients and manufacturing processes of rubber closures with other possible influence on drug products were analyzed.Complied with the requirements of pharmaceutical lifecycle management,the processes of selection and utilization of rubber closures were divided into three periods,including screening,validation and maintenance,and suggestions on the tasks and goals for these three periods were given.Results Though the actual performance of rubber closures was affected by multiple factors,the processability in the pharmaceutical manufacturing processes and the suitability in the periods of storage,transportation and clinical application could be confirmed by reasonable screening and scientific validation of rubber closures.And by the effective quality control of rubber closures,the suitability could be maintained across the lifecycle of drug products.Conclusion By means of the reasonable selection and effective quality control of rubber closures,it would be benefit for keeping drug safety,efficacy and quality controllability throughout the lifecycle.

Objective To interpret the guideline for rubber closures for pharmaceutical packaging in the Chinese Pharmacopoeia(2025 Edition)for relative parties to facilitate their better understanding and reasonable application of the guideline.Methods Based on the considerations of lifecycle risk management and combined with the developing requirements for the quality management of rubber closures,the necessity of establishing the guideline was explained and the relevant contents of quality control were interpreted,and suggestions of the reasonable utilization of rubber closures were given.Results According to the rubber closures'properties and their actual application,relative parties should establish the enterprise standards or the quality agreements of the rubber closures,in line with the general baseline requirements settled by the guideline.Conclusion Based on the risk management concept in the whole process,the established guideline defining the quality control requirements of rubber closures and their critical quality attributes would be helpful to promote the quality control of rubber closures.

ObjectiveTo evaluate the proficiency and consistency of domestic and foreign testing institutions in the field of veterinary drug residue detection in food， and to promote international cooperation and mutual recognition of testing results among these institutions. MethodsA robust statistical analysis was conducted on the testing results of 20 laboratories in eight countries and regions across North America， Europe， and Asia. The laboratories’ testing capabilities were evaluated using Z-score comparison. ResultsAmong the 20 participating laboratories， 18 achieved satisfactory results， resulting in a satisfaction rate of 90%， while 2 laboratories （10%） failed to meet the requirements. The satisfaction rate of domestic laboratories （100%） was higher than that of foreign laboratories （81.8%）. ConclusionDomestic laboratories perform better than overseas laboratories in determining veterinary drug residues in food. To enhance testing capabilities， these overseas laboratories with unsatisfactory evaluation results should strengthen their daily quality control and ensure traceability of original records.

ObjectiveA rapid enrichment and detection method for Escherichia coli O157∶H7 was developed by using multienzyme isothermal rapid amplification （MIRA） fluorescence method combined with metal organic frameworks immunomagnetic beads. MethodsUsing rfbE gene as the target， the primers， probes and reaction system were screened， and the specificity， sensitivity and practical application of this method were investigated. ResultsThe detection limit of Escherichia coli O157∶H7 was 1.18×10⁵ CFU‧mL^-1， and the detection limit of DNA concentration was 9 pg‧μL^-1. The detection process was completed in 20 minutes. The test results of 47 strains （24 target strains and 23 non-target strains） were consistent with real-time PCR （RT-PCR）. ConclusionA method based on metal-organic framework immunomagnetic beads enrichment combined with MIRA assay is developed in this study. The method is simple， rapid and suitable for rapid enrichment and detection of Escherichia coli O157∶H7 in food.

ObjectiveTo establish microbial limit test methods for 44 pediatric drugs. MethodsAccording to the general guidelines in Chinese Pharmacopoeia （2015 and 2020 edition， volume Ⅳ），a suitability test of the methods for 44 drugs was carried out by pour-plate method， neutralization method or dilution method. ResultsTotal aerobic microbial count： chemical oral liquid samples can be tested by 1∶10 plate method；traditional Chinese medicine need to be neutralized firstly. Then oral liquids could be tested by 1∶10 plate method and 1∶100 plate method was used for granules. Total count of molds and yeasts： all the samples can be tested by the 1∶10 plate method. The recoveries of five test strains were between 0.5 and 2.0. The specified microorganisms were all detected in the test group， while not found in the negative control group. ConclusionThe microbial limit test methods for the 44 pediatric drugs are established and the results are reliable and can be used in the quality control．

Objective To investigate the microbial contamination in 54 batches of commercial honey. Methods Aerobic plate colony counts for bacteria and colonies of mould and osmophilic yeasts in honey were determined according to the National Food Safety Standard. The bacteria and fungi in unqualified samples were further identified and analyzed by morphology， MALDI-TOF-MS and large subunit （LSU） rRNA gene sequence. Results Three unqualified batches were found. One batch had aerobic plate colony counts exceeding the standard， with a variety of bacteria including Bacillus sp.， Paenibacillus sp. and Brevibacillus sp. Two batches had mould counts exceeding the standard. Aspergillus sp was detected in both samples （from the same manufacturer）， and the DNA sequence homology was very high， suggesting the mould might come from the same pollution source. Conclusion There are many kinds of microbial contamination in honey. Manufacturers should strengthen the microbe control in monitoring process to avoid the repeated microbial contamination.

Objective To prepare samples of proficiency testing （PT） containing live Staphylococcus aureus with drug matrix in microbial PT program of drug testing， and to evaluate the laboratory capability of microbiology tests in China. Methods Two kinds of PT samples containing living bacteria with drug matrix were prepared and evaluated. The results of laboratories participated in PT program and their response to the survey questionnaire were collected and analyzed. Results The homogeneity and stability of the PT samples complied with the requirements of CNAS-CL03： 2010. Samples could be stored stably for at least 6 months at -20 ℃. Among 63 laboratories participating in PT program， 53 laboratories （84.1%） achieved satisfactory results. The satisfaction rate was 94.1% （16/17） in 17 government laboratories （27.0% of total 63 laboratories）， 81.4% （35/43） in 43 pharmaceutical quality control laboratories （68.3% of total 63 laboratories）， and 66.7% （2/3） in 3 non-government laboratories （4.8% of total 63 laboratories）， respectively. Conclusion The government laboratories performed better than pharmaceutical quality control laboratories in microbiology tests of drug， and the testing abilities of some pharmaceutical laboratories needs to be improved. The preparation and application of microbial samples in drug matrix could provide evaluation tools for drug testing laboratories in microbiology.

OBJECTIVE：To compare the difference of microbiological limit test and criteria of TCM decoction pieces among 43 edition of United States Pharmacopeia （USP43），10.0 edition of European Pharmacopeia （EP10.0），17 edition of Japanese Pharmacopeia （JP17）and 2020 edition of Chinese Pharmacopeia （ChP2020），and to provide refernce for the revision and improvement of microbiological standards for TCM decoction pieces in China. METHODS ：The differences in the microbial enumeration tests method （including sampling and sample preparation ，selection of bacteria and culture medium ，count of microorganisms and heat-resistant bacteria ，etc.），tests for specified microorganisms （including sample pretreatment ，enrichment， separation and identification ，etc.）and microbial related limit criteria were compared among USP 43，EP10.0，JP17 and ChP 2020. RESULTS & CONCLUSIONS ：In terms of microbiological examination of TCM decoction pieces ，USP43，EP10.0，JP17 had their own independent provisions. Chp 2020 added“general rule 1108”. In terms of inspection items ，in addition to the total aerobic bacteria count and total combined yeasts and molds count ，ChP2020 and EP 10.0 provided three methods for the inspection of control bacteria （bile-resistant Gram-negative bacteria ， Escherichia coli ， Salmonella）. On the basis ， JP17 supplemented Staphylococcus aureus test；However，USP43 added Clostridium test method and put forward the concept of objectionable microorganisms risk assessment ；ChP2020 also added a new method for counting heat-resistant bacteria. In terms of microbial limit criteria，USP43 was the most detailed in the classification of TCM decoction pieces ，which was more strict than EP 10.0 and JP 17； ChP2020 had not set up a unified limit for the inspection of control bacteria of TCM decoction pieces. ChP 2020 revised the “microbial limit standard for TCM extracts and TCM decoction pieces ”，but it was not perfect compared with the Pharmacopoeia of the United States ，Europe and Japan. It is suggested that according to the current situation of microbial contamination and control of TCM decoction pieces ，the microbial limit test and criteria of TCM related products in Pharmacopoeia should be gradually improved ，and the microbial limit level of corresponding products should be reasonably refined.

OBJECTIVETo explore the effect of ouabain on intracellular Ca(2+) concentration ([Ca(2+)]i) in thoracic aorta vascular smooth muscle cells (VSMCs) in vitro.

METHODSPrimary SD rat thoracic aorta VSMCs were cultured by tissue adherent method and identified by immunochemistry. The binding ability between ouabain and VSMCs was detected by autoradiography, and fluo 3-AM (a Ca(2+) fluorescent probe) was employed to investigate whether ouabain affected VSMCs within a short period of time. The effect of a truncated fragment of the sodium pump α2 subunit was assayed in antagonizing the effect of ouabain on [Ca(2+)]i in the VSMCs.

RESULTSWithin the concentration range of 0.1-100 nmol/L, ouabain was found to dose-dependently bind to the VSMCs. Different concentrations of ouabain (0-3200 nmol/L) caused a transient, dose-dependent increase in [Ca(2+)]i in the VSMCs, which was antagonized by the application of the truncated fragment of sodium pump α2 subunit.

CONCLUSIONSElevations in [Ca(2+)]i in the VSMCs can be the cytological basis of high ouabain-induced hypertension. The truncated fragment of the sodium pump α2 subunit can antagonize ouabain-induced increase of [Ca(2+)]i in the VSMCs, which provides a clue for understanding the pathogenesis of and devising a therapeutic strategy for high ouabain-induced hypertension.

Animals ; Aorta, Thoracic ; cytology ; Calcium ; metabolism ; Cells, Cultured ; Cytoplasm ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Ouabain ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase