ABSTRACT The rise of antibiotic‐resistant bacteria has intensified global interest in antimicrobial peptides (AMPs) as promising feed additives. Although AMPs were initially considered less prone to resistance due to their broad‐spectrum activity, recent studies have revealed an alarming increase in bacterial resistance to AMPs, though the mechanisms remain poorly understood. In this study, we demonstrate that Staphylococcus aureus can develop stable resistance to the plectasin‐derived AMP NZ2114, as well as nisin and bacitracin, after 35 consecutive days of exposure. Comparative genomic analysis identified five candidate genes associated with resistance, with functional assays revealing significant mutations in ndh (Gln287*), lytD (Ala138Thr), and braS (Asn130Asp) as key contributors. Knockout studies showed that Δ ndh strains exhibited increased resistance to NZ2114, bacitracin, and nisin, alongside reduced intracellular ROS levels and rifampicin mutation rates. In contrast, Δ lytD and Δ braS mutants displayed diminished resistance to NZ2114 and bacitracin, with enhanced biofilm formation in Δ lytD and reduced biofilm capacity in Δ braS . To further investigate these mutations, we generated in situ complementation strains ∆ :: lytD‐ A138T and ∆:: braS‐ N130D, both of which showed heightened resistance compared to wild type, indicating that functional alterations, rather than gene loss, mediate resistance. Notably, resistance phenotypes correlated inversely with bacterial surface anion levels, emphasizing the importance of electrostatic interactions between cationic AMPs and bacterial surface anions in antimicrobial efficacy. These findings provide novel insights into the mechanisms of AMP resistance in S. aureus , highlighting the risk of cross‐resistance and underscoring the need for stringent control of AMP use to mitigate the emergence of resistance.

Staphylococcal cassette chromosome mec (SCCmec) typing is crucial for investigating methicillin-resistant Staphylococcus aureus (MRSA), relying primarily on the combination of ccr and mec gene complexes. To date, 19 ccr genes and 10 ccr gene complexes have been identified, forming 15 SCCmec types. With the vast release of bacterial genome sequences, mining the database for novel ccr gene complexes and SCC/SCCmec elements could enhance MRSA epidemiological studies. In this study, we identified 12 novel ccr genes (6 ccrA, 3 ccrB, and 3 ccrC) through mining of the National Center for Biotechnology Information (NCBI) database, forming 12 novel ccr gene complexes and 10 novel SCC elements. Overexpression of 5 groups of novel Ccr recombinases (CcrA9B3, CcrA10B1, CcrC3, CcrC4, and CcrC5) in a mutant MRSA strain lacking the ccr gene and extrachromosomal circular intermediate (ciSCC) production significantly promoted ciSCC production, demonstrating their biological activity. This discovery provides an opportunity to advance MRSA epidemiological research and develop database-based bacterial typing methods.

IntroductionDandelion (Pugongying) is one of the most frequently used Chinese herbs for treating lactational mastitis (LM). Pugongying granules, a patented medication primarily comprised of dandelion extract, have been approved by CFDA for LM treatment in China. The aims of this study were to investigate the etiology of LM and the mechanism by which Pugongying granules decrease LM symptoms, with a particular focus on the microbial communities found in breastmilk.MethodsParticipants were recruited from a previously performed randomized controlled trial (Identifier: NCT03756324, ClinicalTrials.gov). Between 2019 and 2020, women diagnosed with unilateral LM at the Beijing University of Chinese Medicine Third Affiliated Hospital were enrolled. In total, 42 paired breastmilk samples from the healthy and affected breasts of the participants were collected. Additionally, 37 paired pre- and post-treatment breastmilk samples from the affected breast were collected from women who received a 3-day course of either Pugongying granules (20 women) or cefdinir (17 women). Clinical outcomes [e.g., body temperature, visual analogue scale (VAS) score for breast pain, the percentage of neutrophils (NE%)] were analyzed pre- and post-treatment, and the breastmilk samples were subjected to 16S rRNA gene sequencing to analyze the alpha and beta diversities and identify significant bacteria. Finally, the relationship between microorganisms and clinical outcomes was analyzed.ResultsThere was no significant difference in fever and pain between the Pugongying group and cefdinir group. The most prevalent bacterial genera in breastmilk were Streptococcus and Staphylococcus. Compared to healthy breastmilk, microbial diversity was reduced in affected breastmilk, and there was a higher relative abundance of Streptococcus. After Pugongying treatment, there was an increase in microbial diversity with significantly higher abundance of Corynebacterium. A negative correlation was found between Corynebacterium, VAS score, and NE%. Treatment with cefdinir did not affect microbial diversity. Taken together, our results show a correlation between LM and reduced microbial diversity, as well as an increased abundance of Streptococcus in affected breastmilk.ConclusionPugongying granules enhanced microbial diversity in breastmilk samples. Given the substantial variation in individual microbiomes, identifying specific species of Streptococcus and Corynebacterium associated with LM may provide additional insight into LM pathogenesis and treatment.

OBJECTIVES A defining feature of MRSA is the SCCmec element. The excision and integration of SCCmec elements are catalysed by Ccr recombinases. Currently, seven ccrA, eight ccrB and two ccrC allotypes have been described. However, there have been no recent reports of a novel Ccr recombinase and thus this area should be explored. METHODS According to the proposed criteria of the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) committee, novel ccr genes were explored by searching the genome of our laboratory staphylococcal strains, which were isolated from bovine mastitis in Northwest China. The biological activity of the novel Ccr recombinases to excise and integrate SCCmec elements was determined. The distribution of the novel ccr genes in staphylococci was conducted by querying the NCBI nr/nt database. RESULTS We report a set of novel Ccr recombinases CcrA8B9, which share nucleotide identities of 46.6%-50.2% and 47.4%-52.8% with the ccrA and ccrB alleles, respectively. We used PCR to show that CcrA8B9 can excise and integrate the SCCmec element. Furthermore, using NCBI BLAST we showed that the ccrA8B9 genes exist in other staphylococcal strains. Unlike the common ccr genes, ccrA8B9 is located outside the SCCmec/SCC element. CONCLUSIONS The novel Ccr recombinases CcrA8B9 can help excise and integrate SCCmec/SCC from the genome and provide a new way to facilitate the transmission of SCCmec/SCC elements among staphylococci.

将中医"扶正以祛邪"理论与微生物动态平衡理论相结合,提出中药可通过调控乳汁菌群和(或)抑制细菌生长,减少抗生素应用,从而治疗急性乳腺炎.临证中以"主证为主,分期论治,精准施治"为治疗原则,淤滞期以通为和、以消为贵,达到或止痛、或散结的目的 ;炎症期或疏肝清胃,或解郁疏肝以截断病情、扭转病势;成脓期透邪托毒、扶正祛邪;溃脓期气血同调、扶助护场;炎症僵块期调补通腠,温消僵块.

Most isolated strains of Staphylococcus sciuri contain mecA1, the evolutionary origin of mecA, but are sensitive to β-lactams (OS-MRSS, oxacillin-susceptible mecA1-positive S. sciuri). In order to improve the efficacy of antibiotic treatment, it is important to clarify whether the resistance of OS-MRSS to β-lactams is an inducible phenotype. In this study, three OS-MRSS strains with oxacillin MIC = 1 μg/ml were isolated from 29 retail pork samples. The resistance of OS-MRSS to β-lactams (MIC > 256 μg/ml) was found to be induced by oxacillin, and the induced resistance was observed to remain stable within a certain period of time. Interestingly, the induced β-lactam resistance was not caused by mecA1, heterogeneous resistance, or any genetic mutation, but mainly due to increased wall teichoic acid (WTA) synthesis that thickened the cell wall. The induced strains also showed slower growth rate, as well as decreased adhesion ability and biofilm thickness. These phenotypes were found to be achieved through altered gene expression in associated pathways, such as the citrate cycle and pentose phosphate pathway. The results challenge the traditional antibiotic sensitivity test. In the presence of β-lactam antibiotics, OS-MRSS that was initially sensitive to β-lactams was observed to gradually develop β-lactam resistance in several days. This often-neglected phenomenon in antibiotic sensitivity tests requires further research attention.

目的 系统评价针刺治疗化疗相关周围神经病变的临床疗效.方法 计算机检索中国知网、万方数据库、维普数据库、PubMed、Cochrane Library中英文数据库,检索时间均从建库至2020年4月20日.由2名独立研究者独立完成文献筛选、资料提取、偏倚风险评价后,采用RevMan5.3和Stata15.0软件进行Meta分析.结果 纳入18项研究,共包括904例患者,其中试验组453例,对照组451例.Meta分析结果显示,针刺能改善患者腓总神经的运动神经传导速度[SMD=0.58,95％CI(0.13,1.03)]及感觉神经传导速度[SMD=0.40,95％CI(0.18,0.61)],并改善癌痛的疼痛数字评分法(NRS)评分[SMD=﹣1.36,95％CI(﹣1.96,﹣0.76)],但两组患者正中神经传导速度的差异无统计学意义(P>0.05).结论 本研究结果表明针刺能有效改善患者腓总神经传导速度,并缓解疼痛症状.但本研究结论仍需更多大样本、高质量的随机对照试验来进一步验证.

目的 探讨哺乳期急性乳腺炎黄腻苔患者和哺乳期健康薄白苔女性的舌苔菌群差异,为预防和治疗急性乳腺炎提供思路.方法 纳入伴有发热、黄腻苔的急性乳腺炎患者组成黄腻苔组,共24例.纳入健康哺乳期薄白苔女性组成薄白苔组,共17例.采集两组受试者舌苔,采用MiSeq高通量测序技术和贝叶斯算法分析两组舌苔样本的菌群组成;采用Studentt检验比较两组舌苔菌群Alpha多样性(包括Chao指数、Shannoneven指数及香农指数);运用偏最小二乘法(PLS-DA)判别两组舌苔菌群结构的相似性;分别采用Wilcoxon秩和检验和LEfSe多级物种差异判别分析法对两组差异物种进行分析.结果 黄腻苔组特有菌门2个,为Deinococcota和Acidobacteriota,特有菌属34个,薄白苔组特有菌属16个,两组舌苔微生物群组成具有明显差异.黄腻苔组Shannoneven指数和香农指数显著高于薄白苔组(P＜0.05).黄腻苔组拟杆菌门含量为29.68％,高于薄白苔组的21.6％,而变形菌门含量为9.43％,低于薄白苔组的29.93％ (P ＜0.05).菌属方面,黄腻苔组普雷沃氏菌属、韦荣球菌属和拟普雷沃氏菌属含量明显高于薄白苔组,而薄白苔组奈瑟氏球菌属和嗜血杆菌属含量明显高于黄腻舌苔(P＜0.05).黄腻苔组主要优势菌属韦荣球菌属、巨球型菌属、棒状杆菌属、普雷沃氏菌属、奇异菌属、拟普雷沃氏菌属和Solobacterium;薄白苔组主要优势菌属是奈瑟氏菌属、嗜血杆菌属、小链菌属,两组组间比较差异均具有统计学意义(P＜0.05).结论 哺乳期急性乳腺炎黄腻苔患者舌苔菌群均匀性和多样性高于哺乳期薄白苔健康女性,且物种差异主要表现为普雷沃氏菌属、拟普雷沃氏菌属和韦荣球菌属丰富度较高.

基于《伤寒杂病论》"三阳合病"观点出发,探讨三阳经辨证论治在哺乳期急性乳腺炎的应用,哺乳期急性乳腺炎是临床常见病,减少抗生素的应用,早期有效阻断病程是治疗难点.三阳合病是正邪交争于太阳、阳明、少阳三阳经的病证.病在三阳,而有三阳之异.哺乳期急性乳腺炎从三阳经辨证论治,首辨某经邪气偏重不同.三阳合病而邪气偏重一经直解该经,偏于太阳宜汗解,偏于阳明宜清下,偏于少阳宜和解.三阳合病而邪气偏重某两经可活用本经方证、化裁合方.三阳合病应留意腑证,予本经方证合用下法.三阳经辨治哺乳期急性乳腺炎,有利于早期截断病程,以防成脓之患.

Abstract • Background: Acute mastitis influences the health condition and quality of life of the infants and mothers during the lactation. Pugongying (a kind of Chinese patent medicine, Herba Taraxaci) has shown benefits in lactating women with acute mastitis in clinical practice. However, there is no solid evidence to support its effectiveness and safety.• Methods: A three-arm, multicenter, randomized, active-controlled, outcome assessor-blinded clinical trial will be undergoing in three hospitals in Beijing. 306 participants will be randomly assigned into three groups in 1:1:1 ratio with Pugongying alone, cefdinir alone, and combination of Pugongying and cefdinir for 3-day intervention drugs administration. And in combination of Pugongying and cefdinir group, the participants will be administrated with 2-day cefdinir and 3-day Pugongying. The primary outcomes are resolution of fever, visual analogue scale (VAS) scores of breast pain, and the size of the breast mass by palpation. The secondary outcomes are the patency of milk, Traditional Chinese Medicine (TCM) symptoms scores, white blood cell count, the percentage of neutrophil and C-reactive protein, relapse at 3th day of follow up after completion of treatment, and safety assessment including routine blood, liver and renal function and electrocardiography. Besides, the incidence of surgery and the quantity of additional intervention drugs will also be evaluated.• Discussion: The results of this trial are expected to confirm whether Chinese herbal medicine Pugongying could alleviate the symptoms and signs in lactating women with acute mastitis, and they could reduce application of cefdinir in clinical practice.• Trial registration: ClinicalTrials.gov [Home - ClinicalTrials.gov], ID: NCT03756324. Registered on December 18th 2018. https://www.clinicaltrials.gov/ct2/show/NCT03756324?cond=Acute+mastitis&draw=2&rank=1

目的:系统评价中药外治法治疗乳腺癌术后上肢淋巴水肿的疗效与安全性.方法:对5个数据库进行检索,检索时间均为从建库至2017年4月,收集所有中医药治疗乳腺癌术后上肢淋巴水肿的随对照试验(RCT).两名研究者独立进行文献筛选、资料提取及严格评价的过程.采用Cochrane偏倚风险评估工具对纳入研究进行方法学质量评价.数据分析运用Revman5.1软件.结果:共纳入15篇文献,1076名患者.Meta分析结果显示,中药外治法联合其他治疗方法治疗乳腺癌术后上肢淋巴水肿在淋巴水肿有效率和减轻患肢疼痛方面优于单独使用其他治疗方法(RR=1.37,95％ CI:[1.25,1.50],P＜0.000 01)、(MD=-0.53,95％ CI:[-0.94,-0.12],P=0.11).上肢周径的分析结果不一致,其中有5项研究的Meta分析结果提示中药外治法联合其他疗法在缩小上肢周径疗效方面优于其他治疗方法.结论:中药外治法治疗乳腺癌术后上肢淋巴水肿可以减轻患者患肢的淋巴水肿程度,并且无明显不良反应.但由于纳入研究的方法学质量不高,该结论有待大样本、高质量的RCT进一步证实.

在浆细胞性乳腺炎的肿块期或脓肿形成之前,内服中药可促进肿块及脓肿消散,配合中药外敷则相得益彰.若患者失治误治,病情迁延至脓肿期或即将破溃,采用中医清创术配合祛腐药线引流,脓清后垫棉绑缚.中医辨证论治的思路贯穿本病治疗的各个方面,对于皮瓣内卷、肤色瘀黑、水肿增厚、创面苍白的“阴性皮瓣”,则不宜急于收口,治疗应刮除水肿肉芽及坏死皮缘,使其成为创面红活、血色鲜红的“阳性皮瓣”后,方可收口.对于瘘管期患者,虽然中医手术扩创配合术后换药是治疗本病的主要手段,但内服中药可起到良好的辅助作用.对于抵触手术治疗的患者,采用疏肝、健脾、补肾分期辨证配合中药外敷治疗本病,可基本摒弃手术切除的治疗方法.虽然中医治疗浆细胞性乳腺炎具有一定优势,但还需要在以下方面突破:①个体化诊治与规范化诊治相结合,优化外治方法,改进术式,兼顾美容效果;②明确并统一中医外治法的诊疗操作规范;③改进研究方法,例如建立浆细胞性乳腺炎中医诊疗数据库;④拓展研究范围,关注中医疗法对患者生存质量改善作用等.希望随着研究的不断深入,能使中医对浆细胞性乳腺炎的治疗更加系统化和规范化.

[目的]系统评价中医药治疗卵巢储备功能减退的临床疗效.[方法]运用循证医学的方法,全面检索从建库至今发表的国内及国外文献库关于中医药治疗卵巢储备功能减退的临床随机对照试验.运用偏倚风险测量工具对纳入文献的质量进行评价.应用RevMan 5.1统计软件对纳入文献的结局指标进行Meta分析.[结果]最终纳入18项临床随机对照研究,分为中药对照西药激素及中药联合西药激素对照西药激素两个组别分别进行研究.同时采用辨证论治进行亚组分析,Meta分析结果显示中药联合西药激素在降低促卵泡生成激素(FSH)、改善中医证候疗效方面效果显著优于西药激素;在降低FSH/促黄体生成激素(LH)、升高抗苗勒管激素(AMH)方面,中药联合西药激素与西药激素的效果,无统计学意义.[结论]中药联合西药激素在改善卵巢储备功能减退方面具有一定的疗效.当然,本研究纳入文献的质量较低,因此,需要大量高质量的随机对照试验予以验证,提高其科学性及客观性.

阳和汤出自《外科证治全生集》,是治疗阴疽的代表方剂.现代医家根据其“化阴布阳”的治疗特点,广泛应用于治疗肿瘤、呼吸、消化、内分泌、神经、风湿免疫、乳腺、周围血管、骨科、皮肤、妇科、儿科等各类疾病,效如桴鼓.文章通过回顾近几年相关文献,对阳和汤治疗各种疾病的临床研究进行总结,为现代医家应用阳和汤提供广阔思路.

海藻玉壶汤是治疗瘿瘤初起的代表方剂，具有化痰软坚、消瘿散结的作用，方中以海藻、昆布、海带为主药，化痰软坚散结，半夏、贝母化痰散结，连翘清热散结，青皮、陈皮行气理气，当归、川芎、独活活血调营，甘草调和诸药，诸药合用使气顺则痰消，血行则结散。以气滞血瘀痰凝为病机，以肿块、结节、增生为主要临床表现，发生在甲状腺、乳腺、男性生殖器、肝胆、肿瘤、血液、皮肤、儿科等疾病，海藻玉壶汤加减治疗这些疾病可以理气活血、化痰软坚，使坚者消，肿者散。因海藻玉壶汤中海藻的药理作用，最常应用于甲状腺疾病，并常配伍穿山甲、夏枯草、牡蛎增强软坚散结之功。对于其中有些疾病病例较少，甚至为个案报道，比如乳房异常发育、慢性淋巴细胞白血病等，有待于进一步研究。