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The vaginal epithelium plays pivotal roles in host defense against pathogen invasion, contributing to the maintenance of an acidic microenvironment within the vaginal lumen through the activity of acid-base transport proteins. However, the precise defense mechanisms of the vaginal epithelium after a bacterial infection remain incompletely understood. This study showed that bacterial lipopolysaccharide (LPS) potentiated net proton efflux by up-regulating the expression of Na + -H + exchanger 1 (NHE1) in vaginal epithelial cells. Pharmacologic inhibition or genetic knockdown of Toll-like receptor-4 and the extracellular signal-regulated protein kinase signaling pathway effectively counteracted the up-regulation of NHE1 and the enhanced proton efflux triggered by LPS in vaginal epithelial cells. In vivo studies revealed that LPS administration led to luminal acidification through the up-regulation of NHE1 expression in the rat vagina. Moreover, inhibition of NHE exhibited an impaired defense against acute bacterial infection in the rat vagina. These findings collectively indicate the active involvement of vaginal epithelial cells in facilitating luminal acidification during acute bacterial infection, offering potential insights into the treatment of bacterial vaginosis., Competing Interests: Disclosure Statement None declared., (Copyright © 2024 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to describe the clinical, histopathological, and immunohistochemical characteristics of MPX and offer meaningful insights into the clinicopathology of MPX. We recruited eight men who had sex with men diagnosed with MPX based on positive results from MPX Virus (MPXV)-specific polymerase chain reaction. Skin biopsies were obtained from four selected lesions, including typical and atypical lesions. Histopathological examinations of atypical solitary ulceration revealed infiltrating inflammatory cells predominantly composed of plasma cells and lymphocytes, forming a "sleeve" around the superficial vessels of the dermis. These features might be misinterpreted as indicative of cutaneous syphilis infection. Meanwhile, typical pustular lesions displayed central necrotic epidermis accompanied by perivascular inflammatory infiltrate dominated by neutrophils, as well as ballooning and reticular degeneration of keratinocytes. Additionally, multinucleated keratinocytes and eosinophilic cytoplasmic inclusions known as Guarnieri's bodies were also observed. Importantly, this study represented the pioneering report on immunohistochemical detection of MPXV A29 and A35 proteins in skin lesions, distinguishing it from previous studies that focused on detecting vaccinia virus protein. The anti-MPXV A29 antibody exhibited robust cytoplasmic staining specifically within affected keratinocytes in both adjacent epidermis and hair follicles, thereby it could contribute to the diagnosis of MPX, especially for cases with atypical skin lesions.

Brucellosis is a zoonosis caused by Brucella, which poses a great threat to human health and animal husbandry. Pathogen surveillance is an important measure to prevent brucellosis, but the traditional method is time-consuming and not suitable for field applications. In this study, a recombinase polymerase amplification-SYBR Green I (RPAS) assay was developed for the rapid and visualized detection of Brucella in the field by targeting BCSP31 gene, a conserved marker. The method was highly specific without any cross-reactivity with other common bacteria and its detection limit was 2.14 × 10 4  CFU/mL or g of Brucella at 40 °C for 20 min. It obviates the need for costly instrumentation and exhibits robustness towards background interference in serum, meat, and milk samples. In summary, the RPAS assay is a rapid, visually intuitive, and user-friendly detection that is highly suitable for use in resource-limited settings. Its simplicity and ease of use enable swift on-site detection of Brucella, thereby facilitating timely implementation of preventive measures., (© 2023. Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i.)

Background: Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. Serum biomarkers play an important role in the early diagnosis and prognosis of HCC. Because a certain percentage of HCC patients are negative for alpha-fetoprotein (AFP) , the diagnosis of AFP-negative HCC is essential to improve the detection rate of HCC., Aim: To establish an effective model for diagnosing AFP-negative HCC based on serum tumour biomarkers., Methods: A total of 180 HCC patients were enrolled in this study. The expression levels of GP73, des-γ-carboxyprothrombin (DCP), CK18-M65, and CK18-M30 were detected by a fully automated chemiluminescence analyser. The variables were selected by logistic regression analysis. Several models were constructed using stepwise backward logistic regression. The performance of the models was compared using the C statistic, integrated discrimination improvement, net reclassification improvement, and calibration curves. The clinical utility of the nomogram was assessed using decision curve analysis (DCA)., Results: The results showed that the expression levels of GP73, DCP, CK18-M65, and CK18-M30 were significantly greater in AFP-negative HCC patients than in healthy controls ( P < 0.001). Multivariate logistic regression analysis revealed that GP73, DCP, and CK18-M65 were independent factors for diagnosing AFP-negative HCC. By comparing the diagnostic performance of multiple models, we included GP73 and CK18-M65 as the model variables, and the model had good discrimination ability (area under the curve = 0.946) and good goodness of fit. The DCA curves indicated the good clinical utility of the nomogram., Conclusion: Our study identified GP73 and CK18-M65 as serum biomarkers with certain application value in the diagnosis of AFP-negative HCC. The diagnostic nomogram based on CK18-M65 combined with GP73 demonstrated good performance and effectively identified high-risk groups of patients with HCC., Competing Interests: Conflict-of-interest statement: We have no financial relationships to disclose., (©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.)

Based on both morphological and molecular evidence, it is confirmed that Alseodaphnopsismaguanensis is conspecific with A.hokouensis . Hence, Alseodaphnopsismaguanensis is treated as a synonym of A.hokouensis here. The conservation status of Alseodaphnopsishokouensis is also re-evaluated according to the IUCN Red List Categories and Criteria in this study., Competing Interests: The authors have declared that no competing interests exist., (Lang Li, Dian-Yang Zou, Ya-Meng Mao, Jie Li.)

Endiandramacrocarpa , a new species of Endiandra (Lauraceae) from Yunnan Province of south-western China, is here described and illustrated, based on morphological evidence. Compared to other Endiandra species occurring in south China and the adjacent regions in Indochina, this species is mainly characterised by its much larger ellipsoidal fruits (up to 11 × 6 cm), as well as glabrous branchlets and puberulent inflorescences., (Dian-yang Zou, Guan-long Cao, Jin-guo Zhang, Lang Li, Jie Li.)

Brucella is the causative agent of brucellosis and can be transmitted to humans through aerosolized particles or contaminated food. Brucella abortus (B. abortus), Brucella melitensis (B. melitensis), and Brucella suis (B. suis) are the most virulent of the brucellae, but the traditional detection methods to distinguish them are time-consuming and require high instrumentation. To obtain epidemiological information on Brucella during livestock slaughter and food contamination, we developed a rapid and sensitive triplex recombinant polymerase amplification (triplex-RPA) assay that can simultaneously detect and differentiate between B. abortus, B. melitensis, and B. suis. Three pairs of primers (B1O7F/B1O7R, B192F/B192R, and B285F/B285R) were designed and screened for the establishment of the triplex-RPA assay. After optimization, the assay can be completed within 20 min at 39°C with good specificity and no cross-reactivity with five common pathogens. The triplex-RPA assay has a DNA sensitivity of 1-10 pg and a minimum detection limit of 2.14 × 104-2.14 × 105 CFU g-1 in B. suis spiked samples. It is a potential tool for the detection of Brucella and can effectively differentiate between B. abortus, B. melitensis, and B. suis S2, making it a useful tool for epidemiological investigations., (© The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.)

Background: Cerebral ischemia/reperfusion (I/R) injury after ischemic stroke is usually accompanied with the activation of inflammasome which seriously impairs neurological function. MiR-139 has been reported to be associated with inflammatory regulation in multiple diseases. However, its effect and mechanism on inflammation regulation after cerebral I/R injury are still poorly understood., Methods: An in vitro model of cerebral I/R injury was constructed with oxygen-glucose deprivation/reoxygenation (OGD/R) treatment. TargetScan bioinformatics analysis and dual luciferase reporter assay were utilized to confirm the targeted relationship between miR-139 and c-Jun. Cell pyroptosis was verified by flow cytometry and Caspase-1 Detection Kit. qRT-PCR assay was performed to detect the expression levels of miR-139, c-Jun, NLRP3 and ASC. Western blotting was applied to measure the protein levels of c-Jun and pyroptosis-related markers NLRP3, ASC, caspase-1, GSDMD Nterm . The ELISA assay was applied to measure the release of IL-1β, IL-18 and LDH., Results: MiR-139 was significantly downregulated whereas c-Jun was obviously upregulated after OGD/R treatment. TargetScan analysis predicted that c-Jun was a potential target of miR-139, which was verified by the dual-luciferase reporter assay. Also, overexpression of miR-139 repressed c-Jun expression. Furthermore, miR-139 inhibited OGD/R-induced cell pyroptosis and the upregulation of NLRP3, caspase-1, ASC, GSDMD Nterm , and the release of IL-1β, IL-18 and LDH, while miR-139 inhibition exerted the opposite effects. However, overexpression of c-Jun aggravated OGD/R-induced nerve injury and partly abolished the neuroprotective effect of miR-139., Conclusion: Upregulation of miR-139 exerted neuroprotection against OGD/R-induced nerve injury by negatively regulating c-Jun/NLRP3 inflammasome signaling. This study offered insights for providing potential therapeutic targets for treating cerebral I/R injury., Competing Interests: Declaration of Competing Interest No authors have competing interests to disclose., (Copyright © 2020 Elsevier Inc. All rights reserved.)

An analytical approach was developed to simultaneously determine 13 antibiotics in sulfonamides, quinolones, and macrolides in leafy vegetables by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-MS/MS). After optimizing extracted solutions, purification methods, and eluents of antibiotics in vegetable substrates, and taking into account the influence of environmental changes and experimental conditions on the results, the optimal experimental scheme was determined. This involved ①weighing 500 mg of vegetable samples and adding 20 mL of methanol-Mcllvaine-Na 2 EDTA solution; ② conducting ultrasonic and centrifugal extraction three times; ③ Allowing rotary evaporation to 20 mL to pass a HLB solid phase extraction column; ④ Eluting the extraction column using 6 mL of methanol, upon which the eluent was dried almost completely; ⑤ Re-dissolving the eluent with a mixed solution of acetonitrile:water (volume ratio of 2:8); ⑥ Detecting by UPLC-MS/MS after centrifugation and filtering. Phase A and B of UPLC-MS/MS used an aqueous solution of 1‰ formic acid and acetonitrile, respectively to conduct gradient elution. Results showed that when the pakchoi spiked at 300 ng·g -1 , the spiked recoveries of 13 antibiotics were 38.05%-96.97%. At 150 ng·g -1 , the spiked recoveries were 34.52%-111.10%. At 50 ng·g -1 , the recoveries of standard addition were 41.75%-107.13%, and the relative deviation (RSD) values were all below 8.68%. The detection limit was 0.4-1 ng·g -1 , and the limit of quantification was 1.5-3 ng·g -1 . This demonstrated good extraction and recovery efficiency on different types of leafy vegetables, and presented a good analytical application effect. The antibiotic residues were detected in four kinds of leafy vegetables in found in markets. The total content ranged from 1.59 ng·g -1 to 32.01 ng·g -1 , and the antibiotic content in samples was calculated by dry weight. The content of antibiotics in pakchoi was the highest, followed by Chinese cabbage, lettuce, and coriander. Among the antibiotics detected, sulfadimidine was the most abundant from the selected leafy vegetables. The content of antibiotics was very low, however the potential health risks caused by long-term consumption could not be ignored.

An artificial diet formulated for continuous rearing of the predator Arma chinensis was inferior to natural prey when evaluated using life history parameters. A transcriptome analysis identified differentially expressed genes in diet-fed and prey-fed A. chinensis that were suggestive of molecular mechanisms underlying the nutritive impact of the artificial diet. Changes in the diet formulation were made based on the transcriptome analysis and tested using life history parameters. The quantity of pig liver, chicken egg, tuna fish, biotin, nicotinamide, vitamin B6, thiamine, riboflavin, vitamin C, L-glutamine, and sucrose was reduced, and wheat germ oil, calcium pantothenate and folic acid were increased. Ecuadorian shrimp was added as a partial substitute for tuna fish. Several parameters improved over six generations, including increased egg viability, and decreased egg and adult cannibalism. Additionally, several parameters declined, including longer developmental times for 2nd-5th instars, and decreased nymphal weights. The improvements in life history parameters support the use of transcriptome analyses to help direct formulation improvements. However, the decline in some parameters suggests that additional information, e.g., proteomic data, may be useful as well to maximize diet formulations.

Poultry husbandry is a very important aspect of the agricultural economy in China. However, chicks are often susceptible to infectious disease microorganisms, such as bacteria, viruses and parasites, causing large economic losses in recent years. In the present study, we isolated an Acinetobacter baumannii strain, CCGGD201101, from diseased chicks in the Jilin Province of China. Regression analyses of virulence and LD50 tests conducted using healthy chicks confirmed that A. baumannii CCGGD201101, with an LD50 of 1.81 (±0.11) × 10(4) CFU, was more virulent than A. baumannii ATCC17978, with an LD50 of 1.73 (±0.13) × 10(7) CFU. Moreover, TEM examination showed that the pili of A. baumannii CCGGD201101 were different from those of ATCC17978. Antibiotic sensitivity analyses showed that A. baumannii CCGGD201101 was sensitive to rifampicin but resistant to most other antibiotics. These results imply that A. baumannii strain CCGGD201101 had both virulence enhancement and antibiotic resistance characteristics, which are beneficial for A. baumannii survival under adverse conditions and enhance fitness and invasiveness in the host. A. baumannii CCGGD20101, with its high virulence and antimicrobial resistance, may be one of the pathogens causing death of diseased chicks., (© 2016 The Societies and John Wiley & Sons Australia, Ltd.)

Objective: To study the expression of CD36 mRNA in granulosa cells (GCs) of patients with polycystic ovary syndrome (PCOS) and the impact of testosterone, insulin and PPARγ agonist rosiglitazone on GCs., Methods: The expression of CD36 mRNA inGCs of patients with PCOS and normal controls were assayed byreal-time PCR.The level of CD36 mRNA after treatment with testosterone, insulin, and rosiglitazone in GCs ofnormal controls were also tested by real-time PCR., Results: (1) The expression of CD36mRNA in the GCs of PCOS was significantly higher than that of the controls (P<0.05). (2) When testosterone concentration was 1 nmol/L, CD36 mRNA increased in the GCs, but there was no significantdifference compared to the blank control, (P>0.05). When testosterone concentration was 10 nmol/L, the expression of CD36 mRNA in the GCs was higher than that in the blank control with significant difference (P<0.05). When insulin concentration was 10 nmol/L, the expression of CD36 mRNA increased but the difference was not statistically significant (P>0.05). When insulin concentration was 100 nmol/L, the expression of CD36mRNA in the GCswas higher than that in the blank control (P=0.05). When rosiglitazone concentration was 1nmol/L, the expression of PPARγ mRNA in GCs were significantly increased compared with the blank control (P<0.05). The expression of CD36 mRNA atrosiglitazone concentrationof 10 nmol/Lwere significantly increased compared to the concentration of 1 nmol/L (P<0.05)., Conclusion: High testosterone and insulin induced the expression of CD36 mRNA.Rosiglitazone increased CD36 mRNA in a dose-related manner in GCs.Increased CD36 mRNA in the GCs of PCOS may be related to the clinical characteristics of PCOS.

We theoretically and experimentally study the radiation of current line source embedded in two-dimensional photonic crystals (PCs) made of ferrite rods. Depending on the symmetry of the PCs, the source radiation working at the band edge of the PCs can be multiple or single beam radiation with narrow beam width. Different ways to change the PC symmetry are proposed. Taking advantage of tunable bandgap of the magnetic PC and by lowering the symmetry of the PC, the radiation direction and its working frequency can be reconfigured by bias magnetic field. Experiments demonstrate the dependence of radiation on the PC symmetry.

The impact of a zoophytogenous, insect-free artificial diet and a secondary prey, pupae of Chinese oak silk moth Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae), on the developmental rate, life history parameters, and fertility was examined for F6, F9, and F12 consecutive generations for domesticated Arma chinensis (Fallou) (Heteroptera: Pentatomidae). This study showed that when fed an insect-free artificial diet during both the nymphal and adult stages, developmental times were prolonged, and fecundity, egg viability, net reproductive rates (R0), and intrinsic rates of increase (rm) declined. As a result, the cost to rear A. chinensis on the artificial diet approached 1.7 times the cost of rearing A. chinensis on pupae of A. pernyi. Future diet improvements should attempt to reduce developmental time, increase fecundity, and egg viability and use less costly nutrient sources., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Yersinia enterocolitica (YE) is a main pathogenic bacterium causing diarrhea and yersiniosis occurs in both developed and developing countries with high incidence. YE in contaminated food is able to survive for a long duration even under cold storage, thereby enhancing the risk of food infection. In this study, a new loop-mediated isothermal amplification (LAMP) method showing the characteristics of simplicity, rapidity, high specificity and sensitivity was established by targeting outL of pathogenic YE. Two inner-primers and outer-primers were designed and LAMP reaction was optimized for Mg2+, betaine, dNTPs and inner primers concentrations, reaction temperature and time. Sensitivity and specificity of the LAMP assay was evaluated using YE genomic DNA and those of 44 different bacteria strains, respectively. Validation of LAMP detection method was by employing meat samples spiked with varying CFU of YE. The optimized LAMP assay was specific, capable of detecting 97 fg of genomic DNA (equivalent to 37 genome copies) of YE (100-fold more sensitive than PCR) and 80 CFU/ml of YE-spiked meat samples based on ethidium bromide stained amplicon bands on agarose gel-electrophoresis and on GelRed fluorescence of the LAMP reaction solution, respectively. This rapid, sensitive and specific LAMP technique should enable application in field inspection of Y. enterocolitica in food.

Objective: To research the bacteriostatic effects of Qingkailing Injection Extract (QKLIE) and combination therapy of Qingkailing Injection (QKLI) and antibiotics on bacteria carrying New Delhi metallo-3-lactamase 1 (NDM-1) blaNDM-1 resistance gene, and to determine their minimal inhibitory concentrations (MIC)., Methods: The antimicrobial experiments of QKLIE (Radix Isatidis, baicalin, gardenia, honeysuckle) and combination therapy of QKLI and antibiotics were performed by using the agar dilution method and K-B method. The MIC was determined from each extract., Results: There were different degrees of inhibitory effects on resistant bacteria carrying blaNDM-1 by extracts from main components of QKLI. Of them, the inhibitory effect of baicalin was the best and the MIC of the resistant bacteria was 0.015 g/mL to WD, 0.020 g/mL to WX, 0. 005 g/mL to WJ, and more than 0.020 g/mL to pGEX-4T-NDM-1/DH5alpha (GST-NDM-1), respectively. The MIC value of each extract was sequenced from high to low as baicalin, honeysuckle, gardenia, and Radix Isatidis. Furthermore, combination therapy of QKLI and antibiotics greatly enhanced the antimicrobial activity of each antibiotics when used alone, showing very obvious antibacterial effects on multidrug resistant bacteria carrying blaNDM-1 gene. Of them, the optimal effects were obtained when combined with penicillins (penicillin G, mezlocillin, piperacillin/ tazobactam, ampicillin/sulbactam), with the antibacterial effects improved by 10 folds. The antibacterial effects of other kinds of antibiotics were improved to some extent. Conclusions QKLIE and combination therapy of QKLI and antibiotics showed better bacteriostatic effects on resistant bacteria carrying blaNDM-1 gene. This study provided theoretical bases for drug development, medication and treatment for super-resistant bacteria carrying blaNDM-1.

A single dose (80 mg/kg) of diethylnitrosamine (DEN) was given orally to castrated female Wistar rats. One week after that one half of the animals were treated with diethylstilbestrol (DES) 3 mg/kg/once a week subcutaneously. The other half of the animals received no any hormone or hormone derivatives. The change of the liver cells in animals treated with DEN alone failed to progress beyond the stage of hepatocellular alterations in foci or neoplastic nodules within 8 months, while most of those animals which received DES treatment after DEN initiation developed hepatocellular carcinomas after 6 months. This result denotes that the DES exerts a definite promotive effect on DEN initiated liver cell carcinogenesis.