Insights from Traditional Chinese Medicine for Restoring Skin Barrier Functions.

The skin barrier is essential for maintaining the body's internal homeostasis, protecting against harmful external substances, and regulating water and electrolyte balance. Traditional Chinese Medicine (TCM) offers notable advantages in restoring skin barrier function due to its diverse components, targets, and pathways. Recent studies have demonstrated that active ingredients in TCM can safely and effectively repair damaged skin barriers, reinstating their proper functions. This review article provides a comprehensive overview of the mechanisms underlying skin barrier damage and explores how the bioactive constituents of TCM contribute to skin barrier repair, thereby offering a theoretical framework to inform clinical practices.

Identification of immunotherapy-related subtypes, characterization of tumor microenvironment infiltration, and development of a prognostic signature in gastric carcinoma.

BACKGROUND: Recent advances in immunotherapy have elicited a considerable amount of attention as viable therapeutic options for several cancer types, the present study aimed to explore the immunotherapy-related genes (IRGs) and develop a prognostic risk signature in gastric carcinoma (GC) based on these genes. METHODS: IRGs were identified by comparing immunotherapy responders and non-responders in GC. Then, GC patients were divided into distinct subtypes by unsupervised clustering method based on IRGs, and the differences in immune characteristics and prognostic stratification between these subtypes were analyzed. An immunotherapy-related risk score (IRRS) signature was developed and validated for risk classification and prognosis prediction based on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) cohorts. Besides, the predictive ability of the IRRS in immunotherapy response was also determined. RESULTS: A total of 63 IRGs were identified, and 371 GC patients were stratified into two molecular subgroups with significantly different prognosis and immune characteristics. Then, an IRRS signature comprised of three IRGs (CENP8, NRP1, and SERPINE1) was constructed to predict the prognosis of GC patients in TCGA cohort. Importantly, external validation in multiple GEO cohorts further confirmed the universal applicability of the IRRS in distinct populations. Furthermore, we found that the IRRS was significantly correlated with patient's responsiveness to immunotherapy, GC patients with low IRRS are more likely to benefit from existing immunotherapy. CONCLUSIONS: The risk score could serve as a robust prognostic biomarker, provide therapeutic benefits for immunotherapy and may be helpful for clinical decision making in GC patients.

Research on the Mechanism and Material Basis of Corn (Zea mays L.) Waste Regulating Dyslipidemia.

Corn (Zea mays L.) is an essential gramineous food crop. Traditionally, corn wastes have primarily been used in feed, harmless processing, and industrial applications. Except for corn silk, these wastes have had limited medicinal uses. However, in recent years, scholars have increasingly studied the medicinal value of corn wastes, including corn silk, bracts, husks, stalks, leaves, and cobs. Hyperlipidemia, characterized by abnormal lipid and/or lipoprotein levels in the blood, is the most common form of dyslipidemia today. It is a significant risk factor for atherosclerosis and can lead to cardiovascular and cerebrovascular diseases if severe. According to the authors' literature survey, corn wastes play a promising role in regulating glucose and lipid metabolism. This article reviews the mechanisms and material basis of six different corn wastes in regulating dyslipidemia, aiming to provide a foundation for the research and development of these substances.

Case Report: Snake Venom Ophthalmia Caused by Cobra Exposure: A Report of 26 Cases.

We report 26 cases of eye injuries resulting from cobra venom sprayed by Naja atra. This accounts for 14.5% of patients (26/173) treated for cobra injuries who presented to the emergency department of a snakebite treatment center in Guangzhou, South China. Pain, blurred vision, lacrimation, photophobia, and foreign body sensation were the most common symptoms, found in 24 patients. Ophthalmic examination revealed eyelid swelling and conjunctival congestion. Eye slit lamp examination showed obvious punctate corneal epithelial defects in four patients. Five patients received an intravenous infusion of antivenom. All patients' eyes were rinsed completely with normal saline after their arrival at the hospital. Prophylactic topical antibiotics were given to all patients. All eyes were cured without long-term sequelae.

Lesbian couples' childbearing experiences using assisted reproductive technology: A netnography study.

OBJECTIVE: This study aimed to explore how Chinese lesbian couples perceived having children through assisted reproductive technology (ART) and its impact on their experiences of family formation. DESIGN: This study adopted netnography to investigate online forum data created by self-identified lesbian couples in relation to assisted reproduction. Summative content analysis was used to analyse data. FINDINGS: Based on data analysis, A luan B huai, in which a lesbian conceives a child using her partner's egg, was seen as the best way to establish a family because it created a sense of symbolic connectedness with the child for both of them. Moreover, lesbian couples also indicated the crucial role that having children plays in maintaining family harmony, despite their opposition to heterosexual family traditions. With the stratification of reproductive tourism, certain groups of lesbians - for instance, those with limited social and cultural capital - might be at a disadvantage in the global setting of reproductive tourism. CONCLUSIONS AND IMPLICATIONS: Lesbian couples valued the benefits of ART in assisting them to achieve their childbearing goals and build a family. Healthcare providers should take the initiative to enhance fertility care by addressing the concerns and unique challenges faced by lesbian populations.

Construction dual vacancies to regulate the energy band structure of ZnIn2S4 for enhanced visible light-driven photodegradation of 4-NP.

Most of the intrinsic photocatalysts with visible light response can only generate one active radical due to the limitation of their band structures, which is immediate cause limiting their photocatalytic degradation performance. In this work, ZnIn2S4 with Zn vacancy and S vacancy (VZn+S-ZnIn2S4) was prepared for the first time. As expected, the VZn+S-ZnIn2S4 exhibits remarkable photocatalytic performance for 4-Nitrophenol (4-NP) degradation under visible light and the apparent rate constant is about 11 times that of pristine ZnIn2S4. The construction of dual vacancies can regulate the energy band structure of the ZnIn2S4, enabling it to generate •OH and •O2- simultaneously. Meanwhile, dual vacancies system can also extremely improve the separation efficiency of carriers. It is worth noting that Zn vacancy and S vacancy can capture photogenerated holes and photogenerated electrons, respectively, which is beneficial for photogenerated carriers to participate in radical generation reactions. In addition, a possible 4-NP degradation pathway was proposed based on HPLC-MS analysis. This work provides a new way to construct photocatalysts for photodegradation of pollutants in wastewater.

Changes in dendritic complexity and spine morphology following BCG immunization in APP/PS1 mice.

Bacillus Calmette - Guerin (BCG) is an immune regulator that can enhance hippocampal synaptic plasticity in rats; however, it is unclear whether it can improve synaptic function in a mouse model with Alzheimer's disease (AD). We hypothesized that BCG plays a protective role in AD mice and investigated its effect on dendritic morphology. The results obtained show that BCG immunization significantly increases dendritic complexity, as indicated by the increased number of dendritic intersections and branch points, as well as the increase in the fractal dimension. Furthermore, the number of primary neurites and dendritic length also increased following BCG immunization, which increased the number of spines and promoted maturation. IFN-γ and IL-4 levels increased, while TNF-α levels decreased following BCG immunization; expression levels of p-JAK2, P-STAT3, SYN, and PSD-95 also increased. Therefore, this study demonstrates that BCG immunization in APP/PS1 mice mitigated hippocampal dendritic spine pathology, especially after the third round of immunization. This effect could possibly be attributed to; changes in dendritic arborization and spine morphology or increases in SYN and PSD-95 expression levels. It could also be related to mechanisms of BCG-induced increases in IFN-γ or IL-4/JAK2/STAT3 levels.

BCG immunization in a mouse model for Alzheimer's disease significantly increased dendritic complexity, as indicated by an increase in the number of dendritic intersections and branch points, as well as an increase in the fractal dimension of hippocampal CA1 neurons.

Superresolution Imaging of Cytoskeletal Networks in Fixed Brain Tissue.

Emerging evidence suggests that neurodegeneration is directly linked to dysfunction of cytoskeleton; however, visualizing the organization of cytoskeletal structures in brain tissues remains challenging due to the limitation of resolution of light microscopy. Superresolution imaging overcomes this limitation and resolves subcellular structures below the diffraction barrier of light (20-200 nm), while retaining the advantages of fluorescent microscopy such as simultaneous visualization of multiple proteins and increased signal sensitivity and contrast. However, superresolution imaging approaches have been largely limited to very thin samples such as cultured cells growing as a single monolayer. Analysis of thicker tissue sections represents a technical challenge due to high background fluorescence and quality of the tissue preservation methods. Among superresolution microscopy approaches, structured illumination microscopy is one of the most compatible methods for analyzing thicker native tissue samples. We have developed a methodology that allows maximal preservation and quantitative analyses of cytoskeletal networks in tissue sections from a rodent brain. This methodology includes a specialized fixation protocol, tissue preparation, and image acquisition procedures optimized for the characterization of subcellular cytoskeletal structures using superresolution with structured illumination microscopy.

Design and Embedded Implementation of Secure Image Encryption Scheme Using DWT and 2D-LASM.

In order to further improve the information effectiveness of digital image transmission, an image-encryption algorithm based on 2D-Logistic-adjusted-Sine map (2D-LASM) and Discrete Wavelet Transform (DWT) is proposed. First, a dynamic key with plaintext correlation is generated using Message-Digest Algorithm 5 (MD5), and 2D-LASM chaos is generated based on the key to obtain a chaotic pseudo-random sequence. Secondly, we perform DWT on the plaintext image to map the image from the time domain to the frequency domain and decompose the low-frequency (LF) coefficient and high-frequency (HF) coefficient. Then, the chaotic sequence is used to encrypt the LF coefficient with the structure of "confusion-permutation". We perform the permutation operation on HF coefficient, and we reconstruct the image of the processed LF coefficient and HF coefficient to obtain the frequency-domain ciphertext image. Finally, the ciphertext is dynamically diffused using the chaotic sequence to obtain the final ciphertext. Theoretical analysis and simulation experiments show that the algorithm has a large key space and can effectively resist various attacks. Compared with the spatial-domain algorithms, this algorithm has great advantages in terms of computational complexity, security performance, and encryption efficiency. At the same time, it provides better concealment of the encrypted image while ensuring the encryption efficiency compared to existing frequency-domain methods. The successful implementation on the embedded device in the optical network environment verifies the experimental feasibility of this algorithm in the new network application.

Anatomical Organization of the Rat Subfornical Organ.

The subfornical organ (SFO) is a sensory circumventricular organ located along the anterodorsal wall of the third ventricle. SFO lacks a complete blood-brain barrier (BBB), and thus peripherally-circulating factors can penetrate the SFO parenchyma. These signals are detected by local neurons providing the brain with information from the periphery to mediate central responses to humoral signals and physiological stressors. Circumventricular organs are characterized by the presence of unique populations of non-neuronal cells, such as tanycytes and fenestrated endothelium. However, how these populations are organized within the SFO is not well understood. In this study, we used histological techniques to analyze the anatomical organization of the rat SFO and examined the distribution of neurons, fenestrated and non-fenestrated vasculature, tanycytes, ependymocytes, glia cells, and pericytes within its confines. Our data show that the shell of SFO contains non-fenestrated vasculature, while fenestrated capillaries are restricted to the medial-posterior core region of the SFO and associated with a higher BBB permeability. In contrast to non-fenestrated vessels, fenestrated capillaries are encased in a scaffold created by pericytes and embedded in a network of tanycytic processes. Analysis of c-Fos expression following systemic injections of angiotensin II or hypertonic NaCl reveals distinct neuronal populations responding to these stimuli. Hypertonic NaCl activates ∼13% of SFO neurons located in the shell. Angiotensin II-sensitive neurons represent ∼35% of SFO neurons and their location varies between sexes. Our study provides a comprehensive description of the organization of diverse cellular elements within the SFO, facilitating future investigations in this important brain area.

MicroRNA-101-3p, MicroRNA-195-5p, and MicroRNA-223-3p in Peripheral Blood Mononuclear Cells May Serve as Novel Biomarkers for Syphilis Diagnosis.

OBJECTIVES: Syphilis is a sexually transmitted disease of global prevalence. Current diagnostic methods lack sensitivity and specificity, which limits the early diagnosis and prognosis of the disease. MiRNAs hold great promise as potential biomarkers for infectious diseases diagnosis. We previously profiled the expression of miRNAs in PBMCs from patients with different stages of syphilis. We aimed to further confirm the miR-101-3p, miR-195-5p, and miR-223-3p expression profiles and evaluate their diagnostic value in syphilis infection. METHODS: The expression levels of PBMC-derived miR-101-3p, miR-195-5p, and miR-223-3p were analyzed in 133 syphilis patients, 18 non-syphilis patients, and 23 healthy controls by RT-qPCR. ROC analysis was used to evaluate the differentiation power of these miRNAs in syphilis diagnosis, while the correlation between the expression of these miRNAs and TRUST titer was also statistically analyzed. RESULTS: These miRNAs were significantly upregulated in syphilis patients in a stage-specific manner. ROC analysis indicated that miR-223-3p was powerful in discriminating between controls and patients with early, primary, secondary, and latent syphilis, as well as serological cure; the miR-195-5p/miR-223-3p panel showed an improved capacity to differentiate between syphilis patients, primary, or serofast-stage syphilis and controls, while the three miRNAs combined showed an improved capacity to differentiate latent syphilis or serological cure from controls. Importantly, miR-101-3p and miR-223-3p singly or jointly could specifically distinguish syphilis from non-syphilis patients. Moreover, TRUST titer was significantly correlated with miR-101-3p expression. CONCLUSIONS: MiR-101-3p, miR-195-5p, and miR-223-3p might singly or jointly be potential diagnostic biomarkers at different stages of syphilis.

Elevated V-ATPase Activity Following PTEN Loss Is Required for Enhanced Oncogenic Signaling in Breast Cancer.

PTEN loss-of-function contributes to hyperactivation of the PI3K pathway and to drug resistance in breast cancer. Unchecked PI3K pathway signaling increases activation of the mechanistic target of rapamycin complex 1 (mTORC1), which promotes tumorigenicity. Several studies have suggested that vacuolar (H+)-ATPase (V-ATPase) complex activity is regulated by PI3K signaling. In this study, we showed that loss of PTEN elevated V-ATPase activity. Enhanced V-ATPase activity was mediated by increased expression of the ATPase H+ transporting accessory protein 2 (ATP6AP2), also known as the prorenin receptor (PRR). PRR is cleaved into a secreted extracellular fragment (sPRR) and an intracellular fragment (M8.9) that remains associated with the V-ATPase complex. Reduced PTEN expression increased V-ATPase complex activity in a PRR-dependent manner. Breast cancer cell lines with reduced PTEN expression demonstrated increased PRR expression. Similarly, PRR expression became elevated upon PTEN deletion in a mouse model of breast cancer. Interestingly, concentration of sPRR was elevated in the plasma of patients with breast cancer and correlated with tumor burden in HER2-enriched cancers. Moreover, PRR was essential for proper HER2 receptor expression, localization, and signaling. PRR knockdown attenuated HER2 signaling and resulted in reduced Akt and ERK 1/2 phosphorylation, and in lower mTORC1 activity. Overall, our study demonstrates a mechanism by which PTEN loss in breast cancer can potentiate multiple signaling pathways through upregulation of the V-ATPase complex. IMPLICATIONS: Our study contributed to the understanding of the role of the V-ATPase complex in breast cancer cell tumorigenesis and provided a potential biomarker in breast cancer.

Unique Organization of Actin Cytoskeleton in Magnocellular Vasopressin Neurons in Normal Conditions and in Response to Salt-Loading.

Magnocellular neurosecretory cells (MNCs) are intrinsically osmosensitive and can be activated by increases in blood osmolality, triggering the release of antidiuretic hormone vasopressin (VP) to promote water retention. Hence, the activity of magnocellular VP neurons is one of the key elements contributing to the regulation of body fluid homeostasis in healthy organisms. Chronic exposure to high dietary salt leads to excessive activation of VP neurons, thereby elevating levels of circulating VP, which can cause increases in blood pressure contributing to salt-dependent hypertension. However, the molecular basis underlying high-salt diet-induced hyperactivation of magnocellular VP neurons remains not fully understood. Previous studies suggest that magnocellular neurosecretory neurons contain a subcortical layer of actin filaments and pharmacological stabilization of this actin network potentiates osmotically-induced activation of magnocellular neurons. Using super-resolution imaging in situ, we investigated the organization of the actin cytoskeleton in rat MNCs under normal physiological conditions and after a chronic increase in blood osmolality following 7 d of salt-loading (SL). We found that, in addition to the subcortical layer of actin filaments, magnocellular VP neurons are endowed with a unique network of cytoplasmic actin filaments throughout their somata. Moreover, we revealed that the density of both subcortical and cytoplasmic actin networks in magnocellular VP neurons is dramatically increased following SL. These results suggest that increased osmo-responsiveness of VP neurons following chronic exposure to high dietary salt may be mediated by the modulation of unique actin networks in magnocellular VP neurons, possibly contributing to elevated blood pressure in this condition.

MicroRNA expression profiling of peripheral blood mononuclear cells associated with syphilis.

BACKGROUND: Treponema pallidum (T. pallidum) infection evokes significant immune responses, resulting in tissue damage. The immune mechanism underlying T. pallidum infection is still unclear, although microRNAs (miRNAs) have been shown to influence immune cell function and, consequently, the generation of antibody responses during other microbe infections. However, these mechanisms are unknown for T. pallidum. METHODS: In this study, we performed a comprehensive analysis of differentially expressed miRNAs in healthy individuals, untreated patients with syphilis, patients in the serofast state, and serologically cured patients. miRNAs were profiled from the peripheral blood of patients obtained at the time of serological diagnosis. Then, both the target sequence analysis of these different miRNAs and pathway analysis were performed to identify important immune and cell signaling pathways. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed for microRNA analysis. RESULTS: A total of 74 differentially regulated miRNAs were identified. Following RT-qPCR confirmation, three miRNAs (hsa-miR-195-5p, hsa-miR-223-3p, hsa-miR-589-3p) showed significant differences in the serofast and serologically cured states (P < 0.05). One miRNA (hsa-miR-195-5p) showed significant differences between untreated patients and healthy individuals. CONCLUSIONS: This is the first study of miRNA expression differences in peripheral blood mononuclear cells (PBMCs) in different stages of T. pallium infection. Our study suggests that the combination of three miRNAs has great potential to serve as a non-invasive biomarker of T. pallium infections, which will facilitate better diagnosis and treatment of T. pallium infections.

MicroRNA-101-3p Downregulates TLR2 Expression, Leading to Reduction in Cytokine Production by Treponema pallidum-Stimulated Macrophages.

Treponema pallidum (Tp) infection-induced immune responses can cause tissue damage. However, the underlying mechanism by which Tp infection induces immune response is unclear. Recent studies suggest a regulatory role of microRNAs in host immunity. We assessed whether microRNAs also have a regulatory role in immune response to Tp infection in vitro. Our results showed that microRNA-101-3p (miR-101-3p) levels were significantly higher in peripheral blood mononuclear cells of patients with primary syphilis and those in the serofast state, whereas toll-like receptor (TLR) 2 levels were higher in patients with syphilis than in healthy controls. In vitro, stimulation of THP-1 cells with Tp increased miR-101-3p expression. Moreover, miR-101-3p reduced expression levels of TLR2 mRNA and protein in THP-1 cells via binding to the 3' untranslated region of TLR2. Likewise, miR-101-3p inhibited production of inflammatory cytokines, including IL-1ß, IL-6, tumor necrosis factor-α, and IL-12, in Tp-stimulated macrophages. IL-1ß and IL-6 mRNA expression levels were reduced by transfection of macrophages with a TLR2-specific small interfering RNA. Conversely, overexpression of TLR2 upregulated cytokine expression. Patients with secondary syphilis exhibited the highest levels of plasma IL-6, which were negatively correlated with miR-101-3p. In conclusion, Tp infection upregulates miR-101-3p expression, which in turn inhibits the TLR2 signaling pathway, leading to reduced cytokine production.

Effects of salt loading on the organisation of microtubules in rat magnocellular vasopressin neurones.

Magnocellular vasopressin (VP) neurones are activated by increases in blood osmolality, leading to the secretion of VP into the circulation to promote water retention in the kidney, thus constituting a key mechanism for the regulation of body fluid homeostasis. However, chronic high salt intake can lead to excessive activation of VP neurones and increased circulating levels of VP, contributing to an elevation in blood pressure. Multiple extrinsic factors, such as synaptic inputs and glial cells, modulate the activity of VP neurones. Moreover, magnocellular neurones are intrinsically osmosensitive, and are activated by hypertonicity in the absence of neighbouring cells or synaptic contacts. Hypertonicity triggers cell shrinking, leading to the activation of VP neurones. This cell-autonomous activation is mediated by a scaffold of dense somatic microtubules, uniquely present in VP magnocellular neurones. Treating isolated magnocellular neurones with drugs modulating microtubule stability modifies the sensitivity of neuronal activation in response to acute hypertonic stimuli. However, whether the microtubule network is altered in conditions associated with enhanced neuronal activation and increased VP release, such as chronic high salt intake, remains unknown. We examined the organisation of microtubules in VP neurones of the supraoptic and paraventricular hypothalamic nuclei (SON and PVN, respectively) of rats subjected to salt-loading (drinking 2% NaCl for 7 days). Using super-resolution imaging, we found that the density of microtubules in magnocellular VP neurones from the SON and PVN was significantly increased, whereas the density and organisation of microtubules remain unchanged in other hypothalamic neurones, as well as in neurones from other brain areas (e.g., hippocampus, cortex). We propose that the increase in microtubule density in magnocellular VP neurones in salt-loading promotes their enhanced activation, possibly contributing to elevated blood pressure in this condition.

V-ATPase-associated prorenin receptor is upregulated in prostate cancer after PTEN loss.

Phosphatase and tensin homolog (PTEN) tumor suppressor protein loss is common in prostate cancer (PCa). PTEN loss increases PI3K/Akt signaling, which promotes cell growth and survival. To find secreted biomarkers of PTEN loss, a proteomic screen was used to compare secretomes of cells with and without PTEN expression. We showed that PTEN downregulates Prorenin Receptor (PRR) expression and secretion of soluble Prorenin Receptor (sPRR) in PCa cells and in mouse. PRR is an accessory protein required for assembly of the vacuolar ATPase (V-ATPase) complex. V-ATPase is required for lysosomal acidification, amino acid sensing, efficient mechanistic target of Rapamycin complex 1 (mTORC1) activation, and ß-Catenin signaling. On PCa tissue microarrays, PRR expression displayed a positive correlation with Akt phosphorylation. Moreover, PRR expression was required for proliferation of PCa cells by maintaining V-ATPase function. Further, we provided evidence for a potential clinical role for PRR expression and sPRR concentration in differentiating low from high Gleason grade PCa. Overall, the current study unveils a mechanism by which PTEN can inhibit tumor growth. Lower levels of PRR result in attenuated V-ATPase activity and reduced PCa cell proliferation.

Changing antimicrobial susceptibility and molecular characterisation of Neisseria gonorrhoeae isolates in Guangdong, China: in a background of rapidly rising epidemic.

The prevalence of Neisseria gonorrhoeae infections has increased rapidly since 2015 in China. Antimicrobial resistance and molecular mobilisation in N. gonorrhoeae are two important factors driving this increasing prevalence. This study explored changes in antimicrobial susceptibility and molecular characteristics of N. gonorrhoeae collected in Guangdong, China (2013-2017). A total of 704 isolates were collected in two cities in Guangdong. MICs of major antimicrobials were determined. Penicillinase-producing N. gonorrhoeae (PPNG) and tetracycline-resistant N. gonorrhoeae (TRNG) were characterised, and N. gonorrhoeae multiantigen sequence typing (NG-MAST) was performed. High resistance to penicillin (68.2%), tetracycline (85.7%) and ciprofloxacin (98.2%) was observed. Spectinomycin, ceftriaxone and azithromycin appeared effective, with susceptibilities of 100%, 96.4% and 90.7%, respectively. Resistance to penicillin decreased significantly from 78.4% to 73.6% and to azithromycin from 11.9% to 3.7%. Total prevalence of PPNG, TRNG and PPNG/TRNG was 25.4%, 33.1% and 13.4%, respectively. Rates of PPNG decreased significantly from 37.3% to 23.9%, TRNG from 50.0% to 31.3%, and PPNG/TRNG from 23.5% to 11.7%. However, the ratio of African-type PPNG increased significantly (18.4% to 64.1%) compared with decreasing Asian-type PPNG (81.6% to 33.3%), and the ratio of American-type TRNG increased significantly (0% to 13.7%) compared with decreasing Dutch-type TRNG (100% to 86.3%). A total of 271 sequence types (STs) were identified by NG-MAST from 380 isolates collected in 2013, 2014 and 2017, with 145 novel STs. African-type PPNG is increasing and replacing Asian-type, and novel STs have emerged. Gonococcal isolates with new genotypes might contribute to the rising gonorrhoea epidemic in this area.

Identification and expression analysis of ceftriaxone resistance-related genes in Neisseria gonorrhoeae integrating RNA-Seq data and qRT-PCR validation.

OBJECTIVES: The aim of the study is to identify ceftriaxone resistance-related genes in Neisseria gonorrhoeae. METHODS: Differences in gene expression were compared between ceftriaxone-susceptible N. gonorrhoeae isolates [minimum inhibitory concentration (MIC)=0.002-0.004mg/L] and isolates with decreased ceftriaxone susceptibility (MIC=0.125-0.5mg/L) using RNA-Seq (RNA sequencing). RESULTS: Total RNA of 10 clinical isolates was used to make libraries and generated an average of 24.07Mb reads per sample; these were assembled into 1871 mRNA genes. Moreover, 21 differentially expressed genes (DEGs) were found between the N. gonorrhoeae isolates with susceptibility and decreased susceptibility to ceftriaxone with a fold change of ≥2 (P<0.05), among which 11 were upregulated and 10 were downregulated. Furthermore, all DEGs were verified by quantitative reverse transcription PCR (qRT-PCR), which detected 25 clinical isolates with decreased ceftriaxone susceptibility and 21 ceftriaxone-susceptible isolates. In addition, seven DEGs revealed relative expression levels by 2-ΔΔCt and showed a statistical significance (P≤0.05). Analysis of Gene Ontology (GO) terms and KEGG pathway for functional enrichment showed that six DEGs were related to the cellular component and one DEG was related to the biosynthesis of antibiotics, and these results might be related to ceftriaxone resistance. CONCLUSIONS: Examining ceftriaxone resistance-related genes in N. gonorrhoeae is necessary owing to the high morbidity and antimicrobial resistance of N. gonorrhoeae, especially its eventual resistance to third-generation extended-spectrum cephalosporins (cefixime and ceftriaxone). Moreover, this report provides a new direction for the study and control of ceftriaxone-resistant N. gonorrhoeae.

Determining the in vitro susceptibility of Neisseria gonorrhoeae isolates from 8 cities in Guangdong Province through an improved microdilution method.

A microdilution method for the antibiotic susceptibility testing of Neisseria gonorrhoeae was established and improved, and the antibiotic resistance of N. gonorrhoeae samples isolated from 8 cities of Guangdong in 2016 was determined. The improved microdilution method was compared with the agar dilution method recommend by the World Health Organization (WHO) Western Pacific Region by testing the susceptibility of 100 clinical N. gonorrhoeae isolates. The essential agreement (EA), categorical agreement (CA), very major error (VME), major error (ME), and minor error (MIE) levels of the two methods were analyzed; the acceptable performance rates were measured as follows: ≥90% for EA or CA, ≤3% for VME or ME, and ≤7% for MIE. The EA, CA, VME, ME, and MIE of each method for 7 antibiotics, penicillin, tetracycline, ciprofloxacin, spectinomycin, ceftriaxone, cefixime, and azithromycin, were 96%-100%, 94%-100%, 0%-3%, 0%-2%, and 0%-6%, respectively. The Wilcoxon signed-rank test results indicated 94%-100% agreement between the 2 methods after excluding off-scale values (P > 0.05). The susceptibility of 634 N. gonorrhoeae strains to the 7 antibiotics above were tested through the microdilution method. The resistant rates of the isolates against ciprofloxacin, tetracycline, penicillin, and azithromycin were 99.8%, 88.3%, 53.8%, and 11%, and the percentages of the isolates with decreased susceptibility to ceftriaxone (minimum inhibitory concentration [MIC] ≥0.125 µg/mL) and cefixime (MIC ≥0.25 µg/mL) were 2.1% and 12%, respectively, in Guangdong. Among 8 cities, Shenzhen had the highest rates of resistance against penicillin (77.8%) and decreased susceptibility against ceftriaxone (5.6%). Zhuhai had the highest rates of decreased susceptibility against cefixime (30.1%), and Jiangmen had the highest azithromycin-resistant isolates (16.8%). The findings from this study indicated that the improved microdilution method is an alternative for testing the antimicrobial susceptibility of N. gonorrhoeae. The resistance rates of N. gonorrhoeae against penicillin, tetracycline, and ciprofloxacin were high. While ceftriaxone, cefixime, and spectinomycin remained effective against N. gonorrhoeae, their effectiveness seemed to be decreasing over time. Azithromycin therapy requires timely susceptibility test results.