Differences in the landscape of colonized microorganisms in different oral potentially malignant disorders and squamous cell carcinoma: a multi-group comparative study.

BACKGROUND: The role of microbes in diseases, especially cancer, has garnered significant attention. However, research on the oral microbiota in oral potentially malignant disorders (OPMDs) remains limited. Our study investigates microbial communities in OPMDs. MATERIALS AND METHODS: Oral biopsies from19 oral leukoplakia (OLK) patients, 19 proliferative verrucous leukoplakia (PVL) patients, 19 oral lichen planus (OLP) patients, and 19 oral lichenoid lesions (OLL) patients were obtained. 15 SCC specimens were also collected from PVL patients. Healthy individuals served as controls, and DNA was extracted from their paraffin-embedded tissues. 2bRAD-M sequencing generated taxonomic profiles. Alpha and beta diversity analyses, along with Linear Discriminant Analysis effect size analysis, were conducted. RESULTS: Our results showed the microbial richness and diversity were significantly different among groups, with PVL-SCC resembling controls, while OLK exhibited the highest richness. Each disease group displayed unique microbial compositions, with distinct dominant bacterial species. Noteworthy alterations during PVL-SCC progression included a decline in Fusobacterium periodonticum and an elevation in Prevotella oris. CONCLUSIONS: Different disease groups exhibited distinct dominant bacterial species and microbial compositions. These findings offer promise in elucidating the underlying mechanisms of this disease.

Development of a Pathomics-Based Model for the Prediction of Malignant Transformation in Oral Leukoplakia.

Accurate prognostic stratification of oral leukoplakia (OLK) with risk of malignant transformation into oral squamous cell carcinoma is crucial. We developed an objective and powerful pathomics-based model for the prediction of malignant transformation in OLK using hematoxylin and eosin (H&E)-stained images. In total, 759 H&E-stained images from multicenter cohorts were included. A training set (n = 489), validation set (n = 196), and testing set (n = 74) were used for model development. Four deep learning methods were used to train and validate the model constructed using H&E-stained images. Pathomics features generated through deep learning combined with machine learning algorithms were used to develop a pathomics-based model. Immunohistochemical staining of Ki67, p53, and PD-L1 was used to interpret the black box of the model. Pathomics-based models predicted the malignant transformation of OLK (validation set area under curve [AUC], 0.899; testing set AUC, 0.813) and significantly identified high-risk and low-risk populations. The prediction performance of malignant transformation from dysplasia grading (validation set AUC, 0.743) was lower than that of the pathomics-based model. The expressions of Ki67, p53, and PD-L1 were correlated with various pathomics features. The pathomics-based model accurately predicted the malignant transformation of OLK and may be useful for the objective and rapid assessment of the prognosis of patients with OLK.

The preliminary exploration of immune microenvironment in oral leukoplakia concomitant with oral submucosal fibrosis: A comparative immunohistochemical study.

BACKGROUND: Oral leukoplakia concomitant with oral submucous fibrosis is a high-risk oral potentially malignant disorder, but little is known about its immune microenvironment. METHODS: Thirty samples of oral leukoplakia concomitant with oral submucous fibrosis, 30 oral leukoplakia samples, and 30 oral submucous fibrosis samples were collected from two hospitals. Immunohistochemistry was performed to analyze expression of T cell biomarkers [CD3, CD4, CD8, and Forkhead box P3 (Foxp3)], a B cell biomarker (CD20), macrophage biomarkers (CD68 and CD163), an immune inhibitory receptor ligand (PD-L1), and Ki-67. RESULTS: The numbers of CD3+ (p < 0.001), CD4+ (p = 0.018), and CD8+ (p = 0.031) cells in oral leukoplakia concomitant with oral submucous fibrosis were less than those in oral leukoplakia. The number of CD4+ cells (p = 0.035) in oral leukoplakia concomitant with oral leukoplakia was higher than that in oral submucous fibrosis. More CD3+ (p < 0.001), CD4+ (p < 0.001), Foxp3+ (p = 0.019), and CD163+ (p = 0.029) cells were found in oral leukoplakia than in oral submucous fibrosis. CONCLUSION: Various levels of immune infiltration were observed among oral leukoplakia concomitant with oral submucous fibrosis, oral leukoplakia, and oral submucous fibrosis. Characterization of the immune microenvironment may contribute to personalized immunotherapy.

Architectural and cytological features of epithelial dysplasia associated with transformation risk.

OBJECTIVES: This study explored associations between histological features of dysplasia and malignant transformation, as well as genomic copy number alterations. MATERIALS AND METHODS: Overall, 201 samples were collected from patients of oral leukoplakia. The associations of dysplastic features with malignant transformation and copy number alterations were investigated by Cox proportional hazards regression analysis and the Mann-Whitney U-test. RESULTS: Eight individual histological features, such as irregular epithelial stratification (p = 0.001), mitoses high in epithelium (p = 0.033), extension of changes along minor gland ducts (p < 0.001), etc., were associated with greater risk of malignant transformation. A model including histological features and age showed good performance for predicting malignant transformation (area under receiver operating characteristic curve: 0.806). Irregular epithelial stratification (p = 0.007), abnormal nuclear shape (p = 0.005), abnormal cell size (p = 0.004), etc. were associated with greater genomic instability. CONCLUSIONS: A Cox proportional hazards model using eight histological features and patient age reliably predicted the malignant potential of oral epithelial dysplasia. Identification of these histological features closely related to malignant transformation may aid the management of oral potentially malignant disorders and early detection of oral squamous cell carcinoma.

Activity of chlorhexidine acetate in combination with fluconazole against suspensions and biofilms of Candida auris.

OBJECTIVES: As a newly emerging pathogen, Candida auris has spread rapidly and caused a serious invasive infection. Candida auris often appeared high resistance to classical antifungal drugs. Drug combination therapy is emerging as an effective and well-established strategy to relieve drug resistance problems. The objective of present work was to examine the activity of fluconazole in combination with chlorhexidine acetate against Candida auris isolates. METHODS: Antiplanktonic activity was studied using the EUCAST methodology and growth curve assay. Antibiofilm effectiveness was determined by the crystal violet method, checkerboard microdilution assay, scanning electron microscopy, and confocal laser scanning microscopy. RESULTS: The results indicated that the 80% minimal inhibitory concentrations for fluconazole alone against Candida auris were 2-32 mg/L and for chlorhexidine acetate were 2-8 mg/L. The combination of fluconazole with chlorhexidine acetate exhibited synergism with the growth curve assay. In addition, the checkerboard microdilution assay presented that fluconazole was strongly synergistic with chlorhexidine acetate (sFICI <0.1875) in inhibiting the growth of Candida auris biofilms. The scanning electron microscopy and confocal laser scanning microscopy further exhibited the alteration of morphology of the cells and architecture of the biofilms. CONCLUSION: The combination therapy of fluconazole and chlorhexidine acetate provides a new potential strategy for the treatment of clinical Candida auris infection.

Exome sequencing identifies new somatic alterations and mutation patterns of tongue squamous cell carcinoma in a Chinese population.

Tongue squamous cell carcinoma (TSCC) is an aggressive group of tumors characterized by high rates of regional lymph node metastasis and local recurrence. Emerging evidence has revealed genetic variations of TSCC across different geographical regions due to the impact of multiple risk factors such as chewing betel-quid. However, we know little of the mutational processes of TSCC in the Chinese population without the history of chewing betel-quid/tobacco. To explore the mutational spectrum of this disease, we performed whole-exome sequencing of sample pairs, comprising tumors and normal tissue, from 82 TSCC patients. In addition to identifying seven previously known TSCC-associated genes (TP53, CDKN2A, PIK3CA, NOTCH1, ASXL1, USH2A, and CSMD3), the analysis revealed six new genes (GNAQ, PRG4, RP1, ZNF16, NBEA, and PTPRC) that had not been reported previously in TSCC. Our in vitro experiments identified ZNF16 for the first time as a solid tumor associated gene to promote malignancy of TSCC cells. We also identified a microRNA (miR-585-5p) encoded by the 5q35.1 region and characterized it as a tumor suppressor by targeting SOX9. At least one non-silent mutation of genes involved in the 10 canonical oncogenic pathways (Notch, RTK-RAS, PI3K, Wnt, Cell cycle, p53, Myc, Hippo, TGFß, and Nrf2) was found in 82.9% of cases. Collectively, our data extend the spectrum of TSCC mutations and define novel diagnosis markers and potential clinical targets for TSCC. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Identification of disulfiram as a potential antifungal drug by screening small molecular libraries.

OBJECTIVES: Candida albicans and Candida auris strains are common causative species of Candidiasis. The limited number of antifungal drugs and the current situation of resistance to existing antifungals force us to search for new antifungal alternatives. METHODS: In this work, primary screening of small molecule libraries (Metabolism Compound Library and Epigenetics Compound Library) consisting of 584 compounds against Candida albicans SC5314 was performed. The dose-response assays, XTT assays, scanning electron microscopy and confocal laser scanning microscopy were used to confirm the antifungal activities of the selected compounds against Candida strains. RESULTS: Through the primary screening, we identified five compounds (U73122, disulfiram, BSK805, BIX01294, and GSKJ4) that inhibited strains growth ≥ 80% for dose-response assays. Disulfiram was identified as the most potent repositionable antifungal drug with 50% growth inhibition detected at a concentration as low as 1 mg/L. The further results showed the antifungal activity of disulfiram against biofilm formation of Candida strains with a 50% minimum inhibitory concentration ranging from 32 to 128 mg/L. Further observations by scanning electron microscopy and confocal laser scanning microscopy confirmed the destruction of biofilm architecture and the change of biofilm morphology after being exposed to disulfiram. CONCLUSION: The study indicated the potential clinical application of disulfiram as a promising antifungal drug against candidiasis.

Biopsied non-dental plaque-induced gingival diseases in a Chinese population: a single-institute retrospective study.

BACKGROUND: While inflammatory diseases such as gingivitis and periodontitis induced by dental plaque biofilms constitute the majority of gingival lesions, gingiva can also be affected by a variety of diseases with aetiologies different from bacterial biofilms. The aim of this study was to retrospectively analyze the frequency and distribution of non-dental plaque-induced gingival diseases (NDPIGDs) in the Chinese population in a single institute. METHODS: A total of 6859 samples of biopsied gingival diseases during the period 2000-2019 were obtained from the Department of Pathology, Peking University Hospital of Stomatology. Lesions were categorized by histopathological diagnosis, pathological characteristics and the new classification of gingival health and gingival diseases/conditions. Demographic information, such as gender, location, and age, were also analyzed. RESULTS: Among 6859 biopsied NDPIGD samples, the five most frequent diagnoses included oral squamous cell carcinoma (OSCC, n = 2094), fibrous hyperplasia (n = 2026), pyogenic granuloma (n = 478), epithelial dysplasia (n = 477), and epithelial hyperplasia/hyperkeratosis (n = 436). All types could be grouped into nine categories according to their pathological characteristics. The most common biopsied NDPIGDs category was "hyperplastic lesions" (n = 2648, 38.61%), followed by "malignant neoplasms" (n = 2275, 33.17%). The most common diagnosis types in each category were fibrous hyperplasia and OSCC. Of all NDPIGDs, most lesions could be categorized into the new classification of gingival health and gingival diseases/conditions; only 7.07% did not fit the current classification system. CONCLUSIONS: The present study is the first report on the frequency and distribution of biopsied NDPIGDs in a Chinese population. Unlike previous studies, the most prevalent categories were "hyperplastic lesions" and "malignant neoplasms". The proportion of "malignant neoplasms" and "oral potentially malignant disorders" was remarkably higher than in previous researches. Nevertheless, the study provided epidemiological information on many NDPIGDs, which could be useful for future health policies as well as screening programs.

miR-611 promotes the proliferation, migration and invasion of tongue squamous cell carcinoma cells by targeting FOXN3.

OBJECTIVES: The function of miR-611 has not yet been reported. We aimed to investigate the effects of miR-611 on tongue squamous cell carcinoma (TSCC) and the underlying mechanism. MATERIALS AND METHODS: The expression level of miR-611 in TSCC tissues was measured using quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). Cell proliferation, migration and invasion were examined by performing CCK-8, IncuCyte and Transwell assays. Bioinformatics analyses and microarrays were used to screen for target genes, which were verified using a luciferase reporter assay, RT-qPCR and Western blotting. The xenograft model was used to assess the effects of miR-611 in vivo. RESULTS: miR-611 was upregulated in TSCC tissues, which was significantly correlated with TNM stage and negatively associated with the overall survival of patients. In addition, upregulation of miR-611 not only potentiated the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of TSCC cells in vitro, but also promoted tumour growth in vivo. FOXN3 was identified as a candidate target gene of miR-611 and subsequently verified. Finally, miR-611 induced a malignant phenotype of TSCC, which was rescued by overexpression of FOXN3. CONCLUSIONS: Our findings suggest that miR-611 is a novel therapeutic target for TSCC.

PTCH1 alterations are frequent but other genetic alterations are rare in sporadic odontogenic keratocysts.

OBJECTIVE: Odontogenic keratocysts (OKCs) are benign jaw lesions with high growth potential and propensity for recurrence. Our previous study revealed that PTCH1 mutations, which were frequently detected in sporadic OKCs, might be underestimated due to the masking effect of the stromal components within the tested tissues. We aimed to confirm these results in larger scale and further present the unbiased view of the genomic basis of sporadic OKCs except PTCH1. MATERIALS AND METHODS: We analyzed PTCH1 mutations in additional 19 samples. Using whole-exome sequencing (WES), we further characterized the mutational landscape of five sporadic OKC samples lacking PTCH1 mutation and loss of heterozygosity (LOH). RESULTS: Combined with our previously reported 19 cases, thirty of 38 (79%) cases harbored PTCH1 mutations. Through whole-exome sequencing and integrative analysis, 22 novel mutations were confirmed among five PTCH1-negative samples. No recurrent mutations were identified in the WES samples and validation cohort of 10 OKCs. CONCLUSIONS: Our data further confirmed the frequent PTCH1 mutation and other rare genetic alterations in sporadic OKCs, highlighting the central role of SHH signaling pathway. In PTCH1-negative cases, other rare mutations scattered in a subset of OKCs were independent of the SHH pathway. These results suggested that an SHH inhibitor may be effective to treat the majority of OKCs.

CMTM3 inhibits cell growth and migration and predicts favorable survival in oral squamous cell carcinoma.

Downregulation of CKLF-like MARVEL transmembrane domain-containing member 3 (CMTM3) has been reported in a number of human tumors. However, the role of CMTM3 in oral squamous cell carcinoma (OSCC) remains largely unknown. In this study, we showed that the expression of CMTM3 was significantly reduced in OSCC cell lines and primary tumor specimens (P < 0.001). Methylation-specific PCR showed hypermethylation in CMTM3 promoter in a significant proportion of tumor tissues (61 %). The expression of CMTM3 was associated with T stage, lymph node metastasis, tumor node metastasis (TNM) stage, and recurrence of OSCC patients (P < 0.05, n = 201). More importantly, CMTM3 expression was associated with the prognosis of OSCC patients (P < 0.001) and was an independent prognostic factor (hazard ratio = 0.593, 95 % confidence interval, 0.272-1.292; P = 0.039). Overexpression of CMTM3 inhibited the growth and migration of OSCC cells. In vivo experiments also showed that the growth of OSCC xenografts in nude mice was significantly inhibited by CMTM3 overexpression. These findings indicate that downregulation of CMTM3 due to promoter hypermethylation contributed to the proliferation and migration of OSCC cells and suggest that CMTM3 is an independent prognostic factor for the evaluation of the survival of OSCC patients.

CMTM5 exhibits tumor suppressor activity through promoter methylation in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the most common types of malignancies in the head and neck region. CKLF-like MARVEL transmembrane domain-containing member 5 (CMTM5) has been recently implicated as a tumor suppressor gene in several cancer types. Herein, we examined the expression and function of CMTM5 in oral squamous cell carcinoma. CMTM5 was down-regulated in oral squamous cell lines and tumor samples from patients with promoter methylation. Treatment with the demethylating agent 5-aza-2'-deoxycytidine restored CMTM5 expression. In the OSCC cell lines CAL27 and GNM, the ectopic expression of CMTM5-v1 strongly inhibited cell proliferation and migration and induced apoptosis. In addition, CMTM5-v1 inhibited tumor formation in vivo. Therefore, CMTM5 might act as a putative tumor suppressor gene through promoter methylation in oral squamous cell carcinoma.

A novel bearing current signal diagnosis method combining variational modal decomposition and improved random forests.

A new fault diagnosis approach based on bearing current signals is proposed in this paper. First, in view of strong background noise of the current signal, the variational modal decomposition method is applied to decompose the bearing current signal to obtain multiple intrinsic mode functions, and then the intrinsic mode functions are constructed as the input feature vector according to the kurtosis. Second, to avoid the influence of random forest parameters on the random forest classifier, a random forest faulty bearing diagnostic model optimized by the whale algorithm is established. Finally, the accuracy rate and confusion matrix are adopted to evaluate the prediction effects of both established and traditional models. The classification accuracy of the real damaged bearing fault type can reach 95.11%. The fault diagnosis accuracy of manually damaged bearings can reach 93.83%. The results show that the method proposed in this paper has high accuracy and good generalization ability for bearing fault diagnosis.

Unlocking the multifaceted molecular functions and diverse disease implications of lactylation.

In recent years, a significant breakthrough has emerged in biology, the identification of lactylation, a novel post-translational process. This intriguing modification is not limited to a specific class of proteins but occurs across a diverse range, including histones, signalling molecules, enzymes, and substrates. It can exert a broad regulatory role in various diseases, ranging from developmental anomalies and neurodegenerative disorders to inflammation and cancer. Thus, it presents exciting opportunities for exploring innovative treatment approaches. As a result, there has been a recent surge of research interest, leading to a deeper understanding of the molecular mechanisms and regulatory functions underlying lactylation within physiological and pathological processes. Here, we review the detection and molecular mechanisms of lactylation, from biological functions to disease effects, providing a systematic overview of the mechanisms and functions of this post-translational modification.

Digital pathology-based artificial intelligence models for differential diagnosis and prognosis of sporadic odontogenic keratocysts.

Odontogenic keratocyst (OKC) is a common jaw cyst with a high recurrence rate. OKC combined with basal cell carcinoma as well as skeletal and other developmental abnormalities is thought to be associated with Gorlin syndrome. Moreover, OKC needs to be differentiated from orthokeratinized odontogenic cyst and other jaw cysts. Because of the different prognosis, differential diagnosis of several cysts can contribute to clinical management. We collected 519 cases, comprising a total of 2 157 hematoxylin and eosin-stained images, to develop digital pathology-based artificial intelligence (AI) models for the diagnosis and prognosis of OKC. The Inception_v3 neural network was utilized to train and test models developed from patch-level images. Finally, whole slide image-level AI models were developed by integrating deep learning-generated pathology features with several machine learning algorithms. The AI models showed great performance in the diagnosis (AUC = 0.935, 95% CI: 0.898-0.973) and prognosis (AUC = 0.840, 95%CI: 0.751-0.930) of OKC. The advantages of multiple slides model for integrating of histopathological information are demonstrated through a comparison with the single slide model. Furthermore, the study investigates the correlation between AI features generated by deep learning and pathological findings, highlighting the interpretative potential of AI models in the pathology. Here, we have developed the robust diagnostic and prognostic models for OKC. The AI model that is based on digital pathology shows promise potential for applications in odontogenic diseases of the jaw.

ProcGCN: detecting malicious process in memory based on DGCNN.

The combination of memory forensics and deep learning for malware detection has achieved certain progress, but most existing methods convert process dump to images for classification, which is still based on process byte feature classification. After the malware is loaded into memory, the original byte features will change. Compared with byte features, function call features can represent the behaviors of malware more robustly. Therefore, this article proposes the ProcGCN model, a deep learning model based on DGCNN (Deep Graph Convolutional Neural Network), to detect malicious processes in memory images. First, the process dump is extracted from the whole system memory image; then, the Function Call Graph (FCG) of the process is extracted, and feature vectors for the function node in the FCG are generated based on the word bag model; finally, the FCG is input to the ProcGCN model for classification and detection. Using a public dataset for experiments, the ProcGCN model achieved an accuracy of 98.44% and an F1 score of 0.9828. It shows a better result than the existing deep learning methods based on static features, and its detection speed is faster, which demonstrates the effectiveness of the method based on function call features and graph representation learning in memory forensics.

Genomic alterations in oral multiple primary cancers.

Oral squamous cell carcinoma (OSCC) is the predominant type of oral cancer, while some patients may develop oral multiple primary cancers (MPCs) with unclear etiology. This study aimed to investigate the clinicopathological characteristics and genomic alterations of oral MPCs. Clinicopathological data from patients with oral single primary carcinoma (SPC, n = 202) and oral MPCs (n = 34) were collected and compared. Copy number alteration (CNA) analysis was conducted to identify chromosomal-instability differences among oral MPCs, recurrent OSCC cases, and OSCC patients with lymph node metastasis. Whole-exome sequencing was employed to identify potential unique gene mutations in oral MPCs patients. Additionally, CNA and phylogenetic tree analyses were used to gain preliminary insights into the molecular characteristics of different primary tumors within individual patients. Our findings revealed that, in contrast to oral SPC, females predominated the oral MPCs (70.59%), while smoking and alcohol use were not frequent in MPCs. Moreover, long-term survival outcomes were poorer in oral MPCs. From a CNA perspective, no significant differences were observed between oral MPCs patients and those with recurrence and lymph node metastasis. In addition to commonly mutated genes such as CASP8, TP53 and MUC16, in oral MPCs we also detected relatively rare mutations, such as HS3ST6 and RFPL4A. Furthermore, this study also demonstrated that most MPCs patients exhibited similarities in certain genomic regions within individuals, and distinct differences of the similarity degree were observed between synchronous and metachronous oral MPCs.

Dynamic changes in the water and volatile compounds of chicken breast during the frying process.

The influence of frying times (0, 2, 4, 6, 8, and 10 min) on the continuous changes in the water distribution and the concentrations of key volatile compounds in chicken breast during the frying process were studied. The fried chicken samples could be distinguished by PCA of E-nose and PLS-DA of GC-MS. A total of 40 volatile compounds were identified by GC-MS, and 28 compounds were verified to be the key compounds after further screening by OAVs. The T22 was increased first and then decreased, while the M22 and M23 in fried chicken were considerably decreased and increased with increasing frying time, respectively. The content of the water and the total peak area of LF-NMR in fried chicken samples during the frying process significantly decreased, and the water was transferred from high to low degrees of freedom. In addition, water content, T21, T22, M22 and L* value were positively correlated with most alcohols and aldehydes, and were negatively correlated with pyrazines, while a*, b*, M23 and all amino acids were positively correlated with pyrazines and were negatively correlated with most alcohols and aldehydes. The results may guide the production processes of fried chicken and help produce high-quality chicken products.

A prognostic model built on amino acid metabolism patterns in HPV-associated head and neck squamous cell carcinoma.

OBJECTIVES: To compare amino acid metabolism patterns between HPV-positive and HPV-negative head and neck squamous cell carcinoma (HNSCC) patients and identify key genes for a prognostic model. DESIGN: Utilizing the Cancer Genome Atlas dataset, we analyzed amino acid metabolism genes, differentiated genes between HPV statuses, and selected key genes via LASSO regression for the prognostic model. The model's gene expression was verified through immunohistochemistry in clinical samples. Functional enrichment and CIBERSORTx analyses explored biological functions, molecular mechanisms, and immune cell correlations. The model's prognostic capability was assessed using nomograms, calibration, and decision curve analysis. RESULTS: We identified 1157 key genes associated with amino acid metabolism in HNSCC and HPV status. The prognostic model, featuring genes like IQCN, SLC22A1, SYT12, and TLX3, highlighted functions in development, metabolism, and pathways related to receptors and enzymes. It significantly correlated with immune cell infiltration and outperformed traditional staging in prognosis prediction, despite immunohistochemistry results showing limited clinical identification of HPV-related HNSCC. CONCLUSIONS: Distinct amino acid metabolism patterns differentiate HPV-positive from negative HNSCC patients, underscoring the prognostic model's utility in predicting outcomes and guiding therapeutic strategies.

A critical review of characteristics of domestic wastewater and key treatment techniques in Chinese villages.

As of 2022, China's rural sewage treatment rate is only approximately 31 %. Rapid rural development has led to higher demand. However, China's rural areas are complex and face many problems, such as uneven economic development, population distribution, and water availability. Long-lasting and low-cost wastewater treatment measures are needed for application in rural areas. The quantity and quality of rural domestic wastewater in China were characterized first. Next, the hot topic of domestic wastewater in Chinese villages was confirmed via bibliometric analysis using CiteSpace, and the treatment technologies for rural domestic wastewater were compared. Specifically, the technical status and challenges of the most common technology in rural domestic wastewater treatment, constructed wetlands, were summarized.