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 investigate whether the characteristics of pathogens mediated by Wolbachia can interfere with Japanese encephalitis virus (JEV) replication and explore the regulatory role of Wolbachia on JEV replication transmitted by Culex mosquitoes. Methods Real-time fluorescence quantitative PCR (qPCR) and RNA fluorescence in situ hybridization (RNA-FISH) were used to detect Wolbachia density in Aa23 (naturally infected with Wolbachia) Aedes albopictus cell and negative control Aa23T Aedes albopictus cells (Wolbachia infection was removed by tetracycline treatment). The plaque assay was conducted to measure the viral titers and cytopathic effect (CPE) in Aa23T and Aa23 cells on days 1 to 8 after JEV (P3 strain) infection. Results qPCR and RNA-FISH results consistently showed that the symbiosis of Wolbachia was negative in Aa23T cell. In Aa23 cells, the copy number of the WSP gene of Wolbachia and the fluorescence signal intensity targeting Wolbachia 16S rDNA increased with cell growth time. In response to JEV infection, Wolbachia prolonged the CPE in viral infected Aa23 cells, which compared to infected Aa23T cells. The plaque assay result has obviously showed that JEV titer in Aa23 cells (106 PFU/mL) was significantly lower than that in Aa23T control cells (108 PFU/mL). Conclusions Wolbachia significantly delays CPE of JEV on cells and inhibits JEV replication in A. albopictus cells. To the best of our knowledge, this is the first report that Wolbachia strongly inhibits JEV infection in mosquito cells. It revealed the role of Wolbachia on inhibition of the viruses that transmitted by Culex mosquitoes. In particular, it provides important experimental data and theoretical basis for application of Wolbachia-based mosquito control technology in prevention and control of JEV.

Abstract： Objective　To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods　In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results　The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions　In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.

To establish the experts consensus on the management of delirium in critically ill patients.A special committee was set up by 15 experts from the Chinese Critical Hypothermia-Sedation Therapy Study Group.Each statement was assessed based on the GRADE (Grading of Recommendations Assessment,Development,and Evaluation) principle.Then the Delphi method was adopted by 36 experts to reassess all the statements.(1) Delirium is not only a mental change,but also a clinical syndrome with multiple pathophysiological changes.(2) Delirium is a form of disturbance of consciousness and a manifestation of abnormal brain function.(3) Pain is a common cause of delirium in critically ill patients.Analgesia can reduce the occurrence and development of delirium.(4) Anxiety or depression are important factors for delirium in critically ill patients.(5) The correlation between sedative and analgesic drugs and delirium is uncertain.(6) Pay attention to the relationship between delirium and withdrawal reactions.(7) Pay attention to the relationship between delirium and drug dependence/ withdrawal reactions.(8) Sleep disruption can induce delirium.(9) We should be vigilant against potential risk factors for persistent or recurrent delirium.(10) Critically illness related delirium can affect the diagnosis and treatment of primary diseases,and can also be alleviated with the improvement of primary diseases.(11) Acute change of consciousness and attention deficit are necessary for delirium diagnosis.(12) The combined assessment of confusion assessment method for the intensive care unit and intensive care delirium screening checklist can improve the sensitivity of delirium,especially subclinical delirium.(13) Early identification and intervention of subclinical delirium can reduce its risk of clinical delirium.(14) Daily assessment is helpful for early detection of delirium.(15) Hopoactive delirium and mixed delirium are common and should be emphasized.(16) Delirium may be accompanied by changes in electroencephalogram.Bedside electroencephalogram monitoring should be used in the ICU if conditions warrant.(17) Pay attention to differential diagnosis of delirium and dementia/depression.(18) Pay attention to the role of rapid delirium screening method in delirium management.(19) Assessment of the severity of delirium is an essential part of the diagnosis of delirium.(20) The key to the management of delirium is etiological treatment.(21) Improving environmental factors and making patient comfort can help reduce delirium.(22) Early exercise can reduce the incidence of delirium and shorten the duration of delirium.(23) Communication with patients should be emphasized and strengthened.Family members participation can help reduce the incidence of delirium and promote the recovery of delirium.(24) Pay attention to the role of sleep management in the prevention and treatment of delirium.(25) Dexmedetomidine can shorten the duration of hyperactive delirium or prevent delirium.(26) When using antipsychotics to treat delirium,we should be alert to its effect on the heart rhythm.(27) Delirium management should pay attention to brain functional exercise.(28) Compared with non-critically illness related delirium,the relief of critically illness related delirium will not accomplished at one stroke.(29) Multiple management strategies such as ABCDEF,eCASH and ESCAPE are helpful to prevent and treat delirium and improve the prognosis of critically ill patients.(30) Shortening the duration of delirium can reduce the occurrence of long-term cognitive impairment.(31) Multidisciplinary cooperation and continuous quality improvement can improve delirium management.Consensus can promote delirium management in critically ill patients,optimize analgesia and sedation therapy,and even affect prognosis.

Objective: To investigate the west nile virus (WNV) infection in Xinjiang, China. Methods: Serum samples were collected from patients with fever and chicken in southern Xinjiang, 2012. The presence of WNV-specific immunoglobulin M (IgM) antibodies and neutralizing antibodies was examined through enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutraization test (PRNT₉₀). Results: A total of 1 712 serum samples of outpatients and inpatients were collected in 8 counties in southern Xinjiang. As a result , 22 samples were positive for WNV IgM antibody and 48 samples were positive for WNV neutralization antibody, among which 21 WNV IgM antibody positive samples and 42 WNV neutralization antibody positive samples were from Jiashi county. Of 383 chicken serum samples collected in 4 counties in southern Xinjiang, only 28 samples were positive for WNV neutralizing antibody, interestingly, all positive chicken serum samples were collected from Jiashi county. Conclusions This study revealed that WNV infection occurred in human and poultry in southern Xinjiang, 2012, mainly in Jiashi county.

Objective To investigate the clinical curative effect of Qingrehuatan method in the treatment of insomnia elderly patients with benzodiazepines dependence.Methods From April 2016 to April 2017,49 insomnia elderly patients with benzodiazepines dependence in the Second People's Hospital of Zhoushan were divided into the observation group(20 cases)and control group (29 cases)according to the different treatment methods.In the gradual reduction at the same time,the control group was treated with trazodone,the observation group was treated with Qingrehuatan method.Before treatment and after treatment for 1course,the revised Pittsburgh sleep quality scale (PSQI),index of insomnia symptom score,TCM symptom score,Zung Anxiety Rating Scale score,Wechsler Adult Intelligence Scale(WAIS－RC)and Wechsler Memory Scale(WMS－RC)score were observed in the two groups. Results After treatment,the rate of withdrawal and discontinuation of the observation group was significantly higher than that of the control group (90.0% vs.68.9%),the difference was statistically significant (χ2＝6.918,P＜0.05).After treatment,the PSQI scores and symptom scores of the two groups were significantly decreased,and the PSQI scores and TCM symptom scores of the observation group improved significantly better than those of the control group[(8.8 ±1.1)points vs.(10.6 ±1.3)points,(2.9 ±0.8)points vs.(4.6 ±1.2)points],the differences were statistically significant(t＝5.063,5.535,all P＜0.05).After treatment,the Zung Anxiety Scale scores of the two groups were significantly lower,and the improvement of the observation group was significantly better than that of the control group[(42.90 ±5.65)points vs.(48.21±5.27)points],the difference was statistically significant(t ＝ 3.366,P＜0.05 ).After treatment,the long －term memory,short －term memory and memory quotient of the two groups were significantly increased,and short－term memory and memory quotient of the observation group improved significantly better than those of the control group[(23.30 ±5.31)points vs.(20.55 ±6.58)points,(76.67 ± 12.70)points vs.(67.36 ±19.13)points],the differences were statistically significant (t＝2.274,2.047,all P＜0.05 ).Conclusion Qingrehuatan method in the treatment of insomnia elderly patients with benzodiazepines depend-ence can improve the possible reduction in the process of stopping drug withdrawal symptoms and memory function, and can improve the symptoms of patients at the same time,and it is worthy of clinical application.

Background Immediately ocular rinsing is a key step for the prevention of eye tissue damage after acute chemical bums.A good ophthalmic rinsing solution can neutralize chemical substance and improve the prognosis of patients.Objective This study attempted to evaluate the therapeutic effect of self-made citric aciddisodium hydrogen phosphate buffer and citric acid-disodium hydrogen phosphate-potassium chloride buffer on corneal chemical burns in mice.Methods Citric acid-disodium hydrogen phosphate buffer solution (solution 1) and citric acid-disodium hydrogen phosphate-potassium chloride buffer solution (solution2) with the pH 7.4 were prepared.One hundred and twenty clean male C57 mice aged 6-8 weeks were randomized to two groups,and filter papers containing 1 mol/L H2SO4 or 0.15 mol/L NaOH were attached to the central corneas of the right eyes to create the acid or alkali burning models.Then the eyes were immediately rinsed by 40 ml solution 1,tap water or solution 2 according to the grouping and the model eyes without rinsing served as the control group.The corneal opacity was examined by slit lamp microscope and scored in 3,7 and 14 days after modeling.The percentages of corneal fluorescein staining,corneal neovascularization and corneal ulcer were analyzed.The study protocol was approved by Experimental Animal Ethic Commission of Second Military Medical University.Results In the corneal acid burning models,the number of eye with corneal opacity scored 1 in the solution 1 group,tap water group and solution 2 group was significantly more than that in the non-rinsing group in 3,7 and 14 days after modeling (all at P＜0.01);In 3 days after modeling,the numbers of eye scored 1 were more in the solution 1 group than those in the tap water group and solution 2 group (x2 =11.000,P =0.001;x2 =4.000,P =0.046).There were no differences in the eye number of different corneal opacity scores in 14 days after acid burning (all at P＞0.05).In 3,7 and 14 days after corneal alkali burning,the number of eyes with corneal opacity scored 1-2 was significantly increased in the solution 1 group,tap water group and solution 2 group compared with non-rinsing group (all at P＜0.01).The percentage of corneal ulcer in the solution 1 group,tap water group and solution 2 group was 7％,27％ and 13％,respectively,which was significantly lower than 73％ in the non-rinsing group (P =0.000,0.027,0.003),and no significant differences were seen in various time points after corneal alkali burning (all at P＞O.05).Corneal neovascularization occurred in 50％ mice in non-rinsing group in 14 days after acid burning.However,no neovascularization was seen in the mice of the solution 1 group,tap water group and solution 2 group in both acid and alkali burning mice.Conclusions Citric aciddisodium hydrogen phosphate buffer (pH 7.4) appears to be an effective emergency rinsing solution for corneal chemical burns and the rinsing solution with or without potassium chloride is not obviously affected to the prognosis of corneal chemical burns in the mice.

Objective To investigate the correlation between the incidence of thyroid carcinoma and TSH, and to provide evidence for clinical application.MethodsSelect 450 cases of patients with thyroid from August 2014 to August 2016 in our hospital as the object of study in this group, the analysis of gender and age of patients, lymph nodes, tumor, laboratory examination results, to explore the relationship between TSH levels and thyroid cancer.ResultsThyroid patients with TSH was significantly higher than that in benign tumor group (P<0.05);the level of TSH and patients with lesion diameter, tumor stage, lymph node metastasis and capsule invasion was significantly correlated (P<0.05), but the level of TSH and age, gender, tumor number, vascular invasion and no significant correlation;TSH the level of thyroid cancer patients were significantly increased (P<0.05).ConclusionThe level of TSH in serum is closely related to the occurrence and development of thyroid cancer, and is closely related to lymph node metastasis and tumor stage, and the higher TSH level can increase the risk of thyroid cancer.

Objective: To study the genome molecular characteristics of Getah virus (SC1210) which isolated in Sichuan province in 2012. Methods: Reverse transcriptase polymerase chain reaction (RT-PCR) was used to identify the isolate and the genome was sequenced by the second Ion Torrent PGM. Computer softwares, including Mega Align and Mega 6, were used to analyze the nucleotide and deduced amino acid sequence, and draw phylogenetic trees. Results: SC1210 was identified as Getah virus. The full genome sequence was 11 690nt, the nucleotide and amino acid homology of the full sequence with other strains were 99.2%-99.7% and 96.5%-99.4%.The capsid protein of SC1210 consisting of 804 nucleotides, encoding 268 amino acids and the full-length of E2 protein, had 1 266 nucleotides, encoding 422 amino acids. The nucleotide homology of the capsid protein and the E2 protein with other strains were 94.9%-99.2% and 94.6%-99.6%, and the amino acid were 97%-99.6% and 97.1%-99.5%. The 3′ UTR of the virus included 402 nucleotides and there were three repeat sequence elements and 19 nucleotides conservation sequence. Conclusions The first GETV isolate SC1210 in Sichuan province has a closer relationship with Yunnan strain YN040 and a far genetic relationship with MM2021.

Objective To investigate the effect of L-carnitine injection on oxidative stress and clinical efficacy in patients with acute anterior ST segment elevated myocardial infarction ( STEMI ) and received primary percutaneous coronary intervention .Methods 70 patients with acute anterior wall STEMI undergoing primary PCI in our hospital were randomly divided into the L-carnitine treatment group and the control group.Patients in both groups received same standardized treatment including aspirin , clopidogrel, statins, ACEI/ARB etc.after PCI.Serum levels of superoxied dismutase ( SOD ) and malondialdehyde ( MDA) were measured at arrival and 7 days after PCI.Laboratory results and echocardiography finding were compared.Results (1) The serum SOD levels in both groups were elevated on day 7 after treatment (P<0.05).The serum SOD level on day 7 in the L-carnitine treatment group was higher than the control group (P<0.05).(2) The serum MDA levels in both groups were lower 7 days after treatment (P<0.05).The serum MDA level in the treatment group was lower than the control group (P<0.05).(3) Echocardiography exam on day 7 showed left ventricular end diastolic diameter ( LVEDd) and left ventricular ejection fraction (LVEF) were more significantly improved compared with the control group (P<0.05).Conclusions For AMI patients receiving primary PCI , L-carnitine injection can increase serum SOD concentration and reduce the production of MDA , showing inhibiting effects on oxidative stress and has protective effect on myocardia .