1.Colonization, drug resistance, and molecular epidemiological characteristics of methicillin-resistant Staphylococcus aureus among dairy farm workers in Xinjiang
Jiguo JIN ; Zhaojie WANG ; Yanggui CHEN ; Xixiao MA ; Wanting XU ; Xingyu WANG ; Xiangnan WEI ; Fan WU ; Xintao DANG ; Xueying XIANG ; Jianyong WU ; Fuye LI
Journal of Environmental and Occupational Medicine 2026;43(2):201-207
Background Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen for both human bloodstream infections and mastitis in cows. However, little attention has been paid to the cross-host transmission of MRSA from cows to high-risk groups in China. Objective To determine the MRSA colonization rates among dairy cows and dairy farm workers in Xinjiang, identify the antibiotic resistance profiles and molecular characteristics of the isolates, and provide scientific evidence for the formulation of targeted infection control strategies. Method A cross-sectional survey combined with laboratory pathogen analysis was conducted. From June to August 2024, large-scale dairy farms in Xinjiang region were selected as study sites. Nasal swabs (n=96) and skin swabs (n=39) were collected from workers, and bovine nasal swab samples (n=109) were collected simultaneously. All samples were subjected to MRSA isolation, cultivation, and identification, followed by antibiotic susceptibility testing to characterize resistance phenotypes. Staphylococcus aureus protein A (Spa) typing was performed to determine strain genotypes and elucidate MRSA colonization rates and molecular epidemiological patterns. Results A total of 35 MRSA strains was successfully isolated from 244 samples. The MRSA colonization rates among dairy farm workers and dairy cows were 20.83% (20/96) and 12.84% (14/109), respectively, with an overall isolation rate of 14.34% (35/244). Among the workers, the nasal colonization rate was 16.67% (16/96), and the skin colonization rate was 12.82% (5/39). One worker exhibited MRSA colonization at multiple body sites. All MRSA strains were resistant to cefoxitin (100%, 35/35). The resistance rates to erythromycin and clindamycin were 42.86% (15/35) and 34.29% (12/35), respectively. Thirteen strains showed a multidrug-resistant phenotype, whereas all strains were susceptible to vancomycin. The MRSA isolates exhibited high genetic diversity, with 13 Spa types identified, among which t441 was the most prevalent (8 strains). Both t441 and t034 types were detected in samples from both the dairy cows and their handlers. These two Spa types also carried and stably inherited specific resistance combinations, including erythromycin–clindamycin–cefoxitin and ciprofloxacin–erythromycin–clindamycin–gentamicin–cefoxitin–tetracycline, and a statistically significant association was also observed between the two resistance profiles and the bacterial types (P < 0.001). In addition, one novel Spa type strain was identified. Conclusion MRSA colonization rates among dairy cows and dairy farm workers in Xinjiang are relatively high, with evidence of multi-site colonization. The isolates exhibit high levels of multidrug resistance and genetic diversity, indicating a potential risk of cross-host transmission.
2.Results and analysis of quality monitoring of radiation health technical services in Jiangsu Province, China
Xingjiang CAO ; Xiaosan XU ; Chunyong YANG ; Yuanyuan ZHOU ; Xiangyong FAN ; Shengri LI ; Jin WANG
Chinese Journal of Radiological Health 2025;34(1):36-40
Objective To assess and understand the service capabilities and existing problems of radiation health technical service institutions in Jiangsu Province, China, and provide a basis for improving in-process and post-process supervision as well as enhancing radiation health technical service capabilities. Methods Thirty radiation health technical service institutions in Jiangsu Province were selected as quality monitoring objects from the National Occupational Health Technical Service Institution Management Information System. Evaluations were conducted using a standardized national assessment checklist, and a comprehensive risk assessment was performed by combining the results of laboratory test capability comparisons. Results The 30 institutions all passed the quality monitoring, with an average score of (76.62 ± 5.07). Comprehensive risk assessment identified 8 (26.67%) high-risk institutions, 22 (73.33%) medium-risk institutions, and 0 (0%) low-risk institutions. Conclusion The overall service quality of radiation health technical service institutions in Jiangsu Province is acceptable. However, further training and supervision are needed to improve technical service capacity and reduce service risks.
3.Analysis of Differential Compounds of Poria cocos Medicinal Materials by Integrated Qualitative Strategy Based on UPLC-Q-Orbitrap-MS
Jiayuan WANG ; Xiaohan FAN ; Xiaoxiao WEI ; Rong CAO ; Jin WANG ; Lei WANG ; Fengqing XU ; Shunwang HUANG ; Deling WU ; Hongsu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):148-156
ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), combined with mass defect filtering(MDF) and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials, with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component, global natural product social molecular network(GNPS) was established, and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection(VIP) value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison, 8 compounds were newly identified by MDF, 28 compounds were newly identified by GNPS, and a total of 102 chemical compounds were identified, including 43 triterpenoids, 16 saccharides, 26 amino acids and peptides, 3 nucleosides, and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen, amino acids and peptides were the most abundant in Poria and cultivated Fushen, carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen, type Ⅲ triterpenoids were more abundant in Poriae Cutis, all four types of triterpenoids were higher in Fushenmu, and type Ⅰ, Ⅱ, and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened, including serine, guanosine, gallic acid, 2-octenal, maltotriose, trametenolic acid, dehydroeburicoic acid, dehydrotrametenolic acid, poricoic acid A, poricoic acid B, poricoic acid E and G, but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials, the types of constituents are generally similar, but their relative contents differed significantly among these medicinal materials, especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS, MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.
4.Analysis of Differential Compounds of Poria cocos Medicinal Materials by Integrated Qualitative Strategy Based on UPLC-Q-Orbitrap-MS
Jiayuan WANG ; Xiaohan FAN ; Xiaoxiao WEI ; Rong CAO ; Jin WANG ; Lei WANG ; Fengqing XU ; Shunwang HUANG ; Deling WU ; Hongsu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):148-156
ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), combined with mass defect filtering(MDF) and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials, with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component, global natural product social molecular network(GNPS) was established, and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection(VIP) value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison, 8 compounds were newly identified by MDF, 28 compounds were newly identified by GNPS, and a total of 102 chemical compounds were identified, including 43 triterpenoids, 16 saccharides, 26 amino acids and peptides, 3 nucleosides, and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen, amino acids and peptides were the most abundant in Poria and cultivated Fushen, carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen, type Ⅲ triterpenoids were more abundant in Poriae Cutis, all four types of triterpenoids were higher in Fushenmu, and type Ⅰ, Ⅱ, and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened, including serine, guanosine, gallic acid, 2-octenal, maltotriose, trametenolic acid, dehydroeburicoic acid, dehydrotrametenolic acid, poricoic acid A, poricoic acid B, poricoic acid E and G, but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials, the types of constituents are generally similar, but their relative contents differed significantly among these medicinal materials, especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS, MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.
5.Components and Brain-protective Effect of Chuanxiong Rhizoma-Paeoniae Radix Rubra in Improving Ischemic Stroke Based on UPLC-Q-TOF-MS
Qizhong JIN ; Jie ZHANG ; Lijuan XIU ; Fan XU ; Lei WANG ; Ning WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):20-29
ObjectiveTo investigate the chemical constituents of Chuanxiong Rhizoma-Paeoniae Radix Rubra(CRPRR) that cross the blood-brain barrier in rats with ischemic stroke, their brain-protective effects, and their impact on inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and pharmacodynamic experiments. MethodsA focal cerebral ischemia-reperfusion injury model was established in rats via the middle cerebral artery occlusion/reperfusion (MCAO/R) method using intraluminal suture. Neurological function was evaluated using behavioral scoring. UPLC-Q-TOF-MS was employed to identify the chemical constituents of CRPRR that crossed the blood-brain barrier and entered the cerebrospinal fluid in MCAO/R model rats. Male Sprague-Dawley rats were randomly divided into six groups: sham operation group, model group, low-, medium-, and high-dose CRPRR groups (1.35, 2.7, 5.4 g·kg-1, respectively), and an edaravone group (5 mg·kg-1), with 12 rats in each group. The sham and model groups received normal saline, while the treatment groups received the respective doses of CRPRR once daily by gavage for three consecutive weeks. The brain-protective effects of CRPRR were assessed using the Longa five-point scoring method, open field test, Morris water maze, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin (HE) staining, and transmission electron microscopy. ResultsNine chemical constituents were identified in the cerebrospinal fluid containing CRPRR, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide. Animal experiment results showed that compared with the sham operation group, the model group exhibited disordered neuronal arrangement, severe vacuolation, nuclear pyknosis, and evident mitochondrial swelling. Chromatin aggregation and peripheralization were also observed. Neurological scores and the number of crossings in the central region were significantly increased (P<0.01), while platform crossings were significantly decreased (P<0.01), and clear infarct areas were present (P<0.01). Serum levels and protein expression of TNF-α, IL-1β, and IL-18 were significantly elevated (P<0.01). Compared with the model group, all dose groups of CRPRR showed marked improvement in neuronal morphology which was close to the normal level, with mitochondrial swelling alleviated and chromatin distribution more uniform. The medium- and high-dose groups significantly reduced neurological scores (P<0.01), while the low-, medium-, and high-dose groups significantly reduced the number of central crossings (P<0.01) and infarct volume (P<0.01), and decreased TNF-α, IL-1β, and IL-18 levels (P<0.05, P<0.01) compared with the model group. Furthermore, the medium- and high-dose groups significantly reduced TNF-α protein expression (P<0.05,P<0.01), and the high-dose group significantly reduced IL-1β and IL-18 protein expression (P<0.01). ConclusionThis study confirmed that CRPRR improves neurological function and alleviates brain tissue damage in MCAO/R rats. Its mechanism may be associated with the downregulation of inflammatory factors TNF-α, IL-1β, and IL-18, as well as the presence of nine active chemical constituents in cerebrospinal fluid, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide, which are closely related to their brain-protective effects.
6.Components and Brain-protective Effect of Chuanxiong Rhizoma-Paeoniae Radix Rubra in Improving Ischemic Stroke Based on UPLC-Q-TOF-MS
Qizhong JIN ; Jie ZHANG ; Lijuan XIU ; Fan XU ; Lei WANG ; Ning WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):20-29
ObjectiveTo investigate the chemical constituents of Chuanxiong Rhizoma-Paeoniae Radix Rubra(CRPRR) that cross the blood-brain barrier in rats with ischemic stroke, their brain-protective effects, and their impact on inflammatory factors including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and pharmacodynamic experiments. MethodsA focal cerebral ischemia-reperfusion injury model was established in rats via the middle cerebral artery occlusion/reperfusion (MCAO/R) method using intraluminal suture. Neurological function was evaluated using behavioral scoring. UPLC-Q-TOF-MS was employed to identify the chemical constituents of CRPRR that crossed the blood-brain barrier and entered the cerebrospinal fluid in MCAO/R model rats. Male Sprague-Dawley rats were randomly divided into six groups: sham operation group, model group, low-, medium-, and high-dose CRPRR groups (1.35, 2.7, 5.4 g·kg-1, respectively), and an edaravone group (5 mg·kg-1), with 12 rats in each group. The sham and model groups received normal saline, while the treatment groups received the respective doses of CRPRR once daily by gavage for three consecutive weeks. The brain-protective effects of CRPRR were assessed using the Longa five-point scoring method, open field test, Morris water maze, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin and eosin (HE) staining, and transmission electron microscopy. ResultsNine chemical constituents were identified in the cerebrospinal fluid containing CRPRR, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide. Animal experiment results showed that compared with the sham operation group, the model group exhibited disordered neuronal arrangement, severe vacuolation, nuclear pyknosis, and evident mitochondrial swelling. Chromatin aggregation and peripheralization were also observed. Neurological scores and the number of crossings in the central region were significantly increased (P<0.01), while platform crossings were significantly decreased (P<0.01), and clear infarct areas were present (P<0.01). Serum levels and protein expression of TNF-α, IL-1β, and IL-18 were significantly elevated (P<0.01). Compared with the model group, all dose groups of CRPRR showed marked improvement in neuronal morphology which was close to the normal level, with mitochondrial swelling alleviated and chromatin distribution more uniform. The medium- and high-dose groups significantly reduced neurological scores (P<0.01), while the low-, medium-, and high-dose groups significantly reduced the number of central crossings (P<0.01) and infarct volume (P<0.01), and decreased TNF-α, IL-1β, and IL-18 levels (P<0.05, P<0.01) compared with the model group. Furthermore, the medium- and high-dose groups significantly reduced TNF-α protein expression (P<0.05,P<0.01), and the high-dose group significantly reduced IL-1β and IL-18 protein expression (P<0.01). ConclusionThis study confirmed that CRPRR improves neurological function and alleviates brain tissue damage in MCAO/R rats. Its mechanism may be associated with the downregulation of inflammatory factors TNF-α, IL-1β, and IL-18, as well as the presence of nine active chemical constituents in cerebrospinal fluid, namely paeoniflorin, senkyunolide F, senkyunolide G, paeonimetabolin Ⅰ, paeoniflorin derivative, senkyunolide H, benzoylpaeoniflorin, senkyunolide A, and ligustilide, which are closely related to their brain-protective effects.
7.Simultaneous Determination of 21 Kinds of Aconitum Alkaloids in Biological Specimens and Herbal Wines Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry
Ju YANG ; Guo-Jun LI ; Xian-Mou FAN ; Rui-Bin ZHAO ; Shao-Ming SU ; Xu-Xian FU ; En-Jin ZHU ; Qi-Lin HUANG ; Yao QIN ; Li-Na LI
Chinese Journal of Analytical Chemistry 2025;53(8):1391-1401,后插1-后插6,封3
A method for simultaneous determination of 21 kinds of Aconitum alkaloids(ATS)in biological specimens and infused liquor using ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was developed.The biological samples were pretreated with methanol-acetonitrile(1∶2,V/V)for protein precipitation,while infused liquors were diluted 100-fold with acetonitrile,followed by centrifugation,and filtration by a 0.22-μm membrane.Chromatographic separation was carried out on an EC-C18 column using gradient elution with the mixture of 10 mmol/L ammonium acetate and 0.2%formic acid as mobile phase A and acetonitrile as mobile phase B.With this method,all the analytes were separated within 9.5 min.The samples were detected in positive ESI mode with dynamic multiple reaction monitoring(MRM)and quantified via external standard calibration.The results showed that the concentrations of the analytes in the range of 2-1000 ng/mL had excellent linearity(R2>0.9992)with the peak area.The developed method was successfully used for detection of 21 kinds of aconitum alkaloids,with limits of detection of 0.5-2 ng/mL,quantification limits of 2-6 ng/mL,intra/inter-day precision≤6.0%,spiked recoveries of 89.4%-100.9%,extraction recoveries of 74.2%-104.4%,and matrix effects ranging from-11.1%to 9.2%in blood/urine.The method was applied to detection of 12 samples from 4 fatal aconite poisoning cases,and all 21 kinds of ATS with total alkaloid concentrations of 0.04-4.18 μg/mL in blood and 154.96-422.83 μg/mL in medicinal liquors were detected.Tissue distribution revealed that the order of concentrations from highest to lowest is as follows:urine(157.22 μg/mL)>gastric contents(51.37 μg/mL)>kidney(21.6 μg/g)>whole blood(4.18 μg/mL)>liver(0.03 μg/g).This method showed many advantages such as simple pretreatment,low detection limits,accurate quantification,broad analyte coverage,and superior anti-interference capability in complex matrices,proving ideal for forensic and toxicological analysis of aconitum alkaloids.
8.Research and prospect of integrated traditional Chinese and western medicine in treatment of bronchiectasis.
Qing MIAO ; Zi YANG ; Bo XU ; Sha-Sha YUAN ; Yu-Chen WEI ; Jin-Zhi ZHANG ; Rui LI ; Chang-Zheng FAN
China Journal of Chinese Materia Medica 2025;50(13):3692-3698
Bronchiectasis(BE) is the third major chronic airway disease, and its incidence rate shows a continuously increasing trend. Bronchiectasis is a highly heterogeneous chronic airway disease. Due to structural alterations, airflow limitation, and mucus hypersecretion, clinical treatment faces many challenges. Particularly, problems including Pseudomonas aeruginosa-dominant drug-resistant bacterial colonization, recurrent infections, airway mucus hypersecretion, and impaired lung function are the most urgent, requiring long-term and personalized treatment and management integrating traditional Chinese and western medicine to prevent the recurrence and continuous progression of the disease. In recent years, both traditional Chinese medicine and western medicine have made certain progress in pathogenesis theories, clinical studies, and basic research regarding the therapeutic challenges of bronchiectasis. Therefore, this paper summarized relevant research from the past 10 years and explored future directions and potential advantages of integrated traditional Chinese and western medicine treatment, providing references for optimizing the clinical management strategies for bronchiectasis.
Bronchiectasis/drug therapy*
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Humans
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Drugs, Chinese Herbal/therapeutic use*
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Medicine, Chinese Traditional/methods*
;
Animals
9.Impact of human papillomavirus and coinfection with other sexually transmitted pathogens on male infertility.
Xin FAN ; Ya XU ; Li-Feng XIANG ; Lu-Ping LIU ; Jin-Xiu WAN ; Qiu-Ting DUAN ; Zi-Qin DIAN ; Yi SUN ; Ze WU ; Yun-Hua DONG
Asian Journal of Andrology 2025;27(1):84-89
This study primarily aimed to investigate the prevalence of human papillomavirus (HPV) and other common pathogens of sexually transmitted infections (STIs) in spermatozoa of infertile men and their effects on semen parameters. These pathogens included Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae, Pseudomonas aeruginosa , and Staphylococcus aureus . A total of 1951 men of infertile couples were recruited between 23 March 2023, and 17 May 2023, at the Department of Reproductive Medicine of The First People's Hospital of Yunnan Province (Kunming, China). Multiplex polymerase chain reaction and capillary electrophoresis were used for HPV genotyping. Polymerase chain reaction and electrophoresis were also used to detect the presence of other STIs. The overall prevalence of HPV infection was 12.4%. The top five prevalent HPV subtypes were types 56, 52, 43, 16, and 53 among those tested positive for HPV. Other common infections with high prevalence rates were Ureaplasma urealyticum (28.3%), Ureaplasma parvum (20.4%), and Enterococcus faecalis (9.5%). The prevalence rates of HPV coinfection with Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae , and Staphylococcus aureus were 24.8%, 25.4%, 10.6%, 6.4%, 2.4%, 7.9%, 5.9%, 0.9%, and 1.3%, respectively. The semen volume and total sperm count were greatly decreased by HPV infection alone. Coinfection with HPV and Ureaplasma urealyticum significantly reduced sperm motility and viability. Our study shows that coinfection with STIs is highly prevalent in the semen of infertile men and that coinfection with pathogens can seriously affect semen parameters, emphasizing the necessity of semen screening for STIs.
Humans
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Male
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Infertility, Male/epidemiology*
;
Coinfection/microbiology*
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Papillomavirus Infections/virology*
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Adult
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Sexually Transmitted Diseases/complications*
;
China/epidemiology*
;
Staphylococcus aureus/isolation & purification*
;
Chlamydia trachomatis/isolation & purification*
;
Prevalence
;
Mycoplasma genitalium/isolation & purification*
;
Ureaplasma urealyticum/isolation & purification*
;
Neisseria gonorrhoeae/isolation & purification*
;
Enterococcus faecalis/isolation & purification*
;
Streptococcus agalactiae/isolation & purification*
;
Herpesvirus 2, Human/genetics*
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Pseudomonas aeruginosa/isolation & purification*
;
Semen/virology*
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Sperm Motility
;
Spermatozoa/microbiology*
;
Human Papillomavirus Viruses
10.High-dose estrogen impairs demethylation of H3K27me3 by decreasing Kdm6b expression during ovarian hyperstimulation in mice.
Quanmin KANG ; Fang LE ; Xiayuan XU ; Lifang CHEN ; Shi ZHENG ; Lijun LOU ; Nan JIANG ; Ruimin ZHAO ; Yuanyuan ZHOU ; Juan SHEN ; Minhao HU ; Ning WANG ; Qiongxiao HUANG ; Fan JIN
Journal of Zhejiang University. Science. B 2025;26(3):269-285
Given that ovarian stimulation is vital for assisted reproductive technology (ART) and results in elevated serum estrogen levels, exploring the impact of elevated estrogen exposure on oocytes and embryos is necessary. We investigated the effects of various ovarian stimulation treatments on oocyte and embryo morphology and gene expression using a mouse model and estrogen-treated mouse embryonic stem cells (mESCs). Female C57BL/6J mice were subjected to two types of conventional ovarian stimulation and ovarian hyperstimulation; mice treated with only normal saline served as controls. Hyperstimulation resulted in high serum estrogen levels, enlarged ovaries, an increased number of aberrant oocytes, and decreased embryo formation. The messenger RNA (mRNA)-sequencing of oocytes revealed the dysregulated expression of lysine-specific demethylase 6b (Kdm6b), which may be a key factor indicating hyperstimulation-induced aberrant oocytes and embryos. In vitro, Kdm6b expression was downregulated in mESCs treated with high-dose estrogen; treatment with an estrogen receptor antagonist could reverse this downregulated expression level. Furthermore, treatment with high-dose estrogen resulted in the upregulated expression of histone H3 lysine 27 trimethylation (H3K27me3) and phosphorylated H2A histone family member X (γ-H2AX). Notably, knockdown of Kdm6b and high estrogen levels hindered the formation of embryoid bodies, with a concomitant increase in the expression of H3K27me3 and γ-H2AX. Collectively, our findings revealed that hyperstimulation-induced high-dose estrogen could impair the demethylation of H3K27me3 by reducing Kdm6b expression. Accordingly, Kdm6b could be a promising marker for clinically predicting ART outcomes in patients with ovarian hyperstimulation syndrome.
Female
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Mice
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Demethylation/drug effects*
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Embryonic Stem Cells
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Estrogens/administration & dosage*
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Gene Expression/drug effects*
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Histones/metabolism*
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Jumonji Domain-Containing Histone Demethylases/metabolism*
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Mice, Inbred C57BL
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Oocytes
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Ovary/drug effects*
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Reproductive Techniques, Assisted
;
Animals

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