Objective To examine the impact of extreme temperature and drought stress on the survival of Bulinus globosus, so as to provide the theoretical evidence for the genomic research of Bulinus in absence of reference genes. Methods B. globosus snail samples were collected from Kiwani Shehia in Pemba Island, Zanzibar, Tanzania, and offspring snails were obtained through laboratory breeding and reproduction. A total of 120 10-week-old B. globosus snails from the same generation were selected and randomly assigned into four groups, including the high-temperature drought (HD) group, normal temperature drought (D) group, low-temperature drought (LD) group, and the control (C) group, of 30 snails in each group. Snails in HD, D, and LD groups were placed in beakers containing dry soil at the bottom and subsequently housed in climate chambers at 35, 26 ℃, and 10 ℃, respectively, while snails in Group C were maintained in 500 mL petri dishes containing dechlorinated tap water at 26 ℃. Following 3 days of breeding, living snails in each group were collected, and soft tissues were dissected and isolated. Total RNA was extracted from snail soft tissues for library construction, followed by high-throughput sequencing on the Illumina HiSeq 4000 sequencing system. De novo transcriptome assembly was performed using the Trinity software, and the longest transcripts were selected as unigenes. Gene functional annotations of unigenes were conducted using the Diamond software against Gene Ontology (GO) knowledgebase, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, NCBI non-redundant (NR) protein sequences database, Protein Family (Pfam) database, and UniProtKB/Swiss-Prot (Swiss-Prot) knowledgebase. GO and KEGG enrichment analyses of differentially expressed genes (DEGs) were performed using the topGO and clusterProfiler software, respectively. In addition, four relevant genes were selected for validation using a real-time quantitative PCR (qRT-PCR) assay to verify the reliability of transcriptome sequencing results. Results Following 3 days of breeding, there were 7, 20, 28, and 30 survival B. globosus snails in HD, LD, D, and C groups, with corresponding survival rates of 23.33% (7/30), 66.67% (20/30), 93.33% (28/30), and 100.00% (30/30), respectively (χ² = 52.72, P < 0.001). De novo transcriptome assembly generated 176 942 unigenes, with annotation rates of 0.98%, 13.49%, 26.46%, 12.48%, and 14.39% against GO knowledgebase, KEGG pathway database, NR protein sequences database, Pfam database, and Swiss-Prot knowledgebase, respectively. There were 33 up-regulated and 72 down-regulated genes in Group D, 483 up-regulated and 815 down-regulated genes in Group HD, and 245 up-regulated and 172 down-regulated genes in Group LD relative to in Group C. Following removal of overlapping genes across groups and unmatched genes, 11 candidate genes were identified. GO and KEGG analyses revealed 3 heat shock protein (HSP)-related DEGs in these 11 candidate genes, which were annotated as HSP12.2, HSP70, and HSP20 genes and were all significantly up-regulated in each treatment group. Three immune and nervous system-related DEGs were identified, and were all significantly down-regulated in each treatment group, which were involved in the neural cell adhesion molecule L1-like protein pathway, fibrinogen binding protein pathway, and leukocyte elastase inhibitor-like protein pathway. qRT-PCR assay quantified that the expression trends of four genes related to temperature and drought stress across different treatment groups were highly consistent with transcriptome sequencing data. Conclusion The survival rate of B. globosus significantly reduces under combined stresses of extreme temperature and drought, possibly due to an imbalance in its cellular homeostasis regulatory system.

OBJECTIVE To investigate the effect and mechanism of Jingangteng capsules in the treatment of non-alcoholic fatty liver disease （NAFLD）. METHODS Thirty-two SD rats were randomly divided into normal group and modeling group. The modeling group was fed a high-fat diet to establish a NAFLD model. The successfully modeled rats were then randomly divided into model group， atorvastatin group[positive control， 2 mg/（kg·d）]， and Jingangteng capsules low- and high-dose groups [0.63 and 2.52 mg/（kg·d）]， with 6 rats in each group. The pathological changes of the liver were observed by hematoxylin-eosin staining and oil red O staining. Enzyme-linked immunosorbent assay was performed to determine the serum levels of triglycerides （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine transaminase （ALT）， aspartate transaminase （AST）， tumour necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-6， IL-18. 16S rDNA amplicon sequencing and metabolomics techniques were applied to explore the effects of Jingangteng capsules on gut microbiota and metabolisms in NAFLD rats. Based on the E-mail：591146765@qq.com metabolomics results， Western blot analysis was performed to detect proteins related to the nuclear factor kappa-B （NF-κB）/NOD-like receptor family protein 3 （NLRP3） signaling pathway in the livers of NAFLD rats. RESULTS The experimental results showed that Jingangteng capsules could significantly reduce the serum levels of TG， TC， LDL-C， AST， ALT， TNF-α， IL-1β， IL-6， IL-18， while increased the level of HDL-C， and alleviated the hepatic cellular steatosis and inflammatory infiltration in NAFLD rats. They could regulate the gut microbiota disorders in NAFLD rats， significantly increased the relative abundance of Romboutsia and Oscillospira， and significantly decreased the relative abundance of Blautia （P＜0.05）. They also regulated metabolic disorders primarily by affecting secondary bile acid biosynthesis， fatty acid degradation， O-antigen nucleotide sugar biosynthesis， etc. Results of Western blot assay showed that they significantly reduced the phosphorylation levels of NF-κB p65 and NF-κB inhibitor α， and the protein expression levels of NLRP3， caspase-1 and ASC （P＜0.05 or P＜0.01）. CONCLUSIONS Jingangteng capsules could improve inflammation， lipid accumulation and liver injury in NAFLD rats， regulate the disorders of gut microbiota and metabolisms， and inhibit NF-κB/NLRP3 signaling pathway. Their therapeutic effects against NAFLD are mediated through the inhibition of the NF-κB/NLRP3 signaling pathway.

Objective: To evaluate the efficacy and safety of the single-use hemoperfusion device (UA230) in treating refractory gout (RG) via plasma perfusion. Methods: Thirty-four RG patients aged 18-65 years were recruited and randomly divided into a control group (febuxostat therapy, n=17) and an experimental group (plasma perfusion combined with febuxostat therapy, n=17). Differences in serum uric acid (SUA) levels and urate-lowering rates between the two groups were analyzed using t-tests. Between-group differences in incidence of adverse reactions were analyzed using chi-square tests. Results: At 7 and 14 days post-treatment, SUA levels in the experimental group were significantly lower than those in the control group, with a higher urate-lowering rate (all P<0.05). However, no statistically significant differences in SUA levels or urate-lowering rate were observed at 28 days post-treatment (all P>0.05). The incidence of adverse reactions showed no significant difference between the two groups (χ =0.15, P>0.05). Conclusion: The single-use hemoperfusion device (UA230), combined with plasma perfusion technology, is a safe and effective treatment for RG. It may serve as a novel therapeutic approach for RG patients in clinical practice.

Objective : To construct a human keratinocyte-forming cell line(HaCaT) with stable knockout of the G protein-coupled receptor 107(GPR107) gene, and to preliminarily investigate the effect of GPR107 deletion on the biological behaviour of HaCaT cells. Methods : Using CRISPR/Cas9 gene editing technology, HaCaT cells with knockout ofGPR107gene were constructed and monoclonal cells with GPR107 deletion were obtained by limited dilution method. Genomic DNA was amplified using Western blot and PCR and sequenced to validate the single-cell clones with knockdown of GPR107. The cell cycle changes were detected by flow cytometry; cell proliferation was detected by CCK-8; apoptosis was detected by flow cytometry; changes in cell differentiation markers were detected by Western blot; cell migration ability was analyzed by cell scratch assay and other methods. Results : LentiCas9-Blast and plenti-guide-RNA-GPR107 plasmids were successfully transfected into HaCaT cells, 21 monoclonal cell lines were obtained by limited dilution, and Western blot showed that the GPR107 expression was significantly reduced in 8 of them; PCR sequencing of the cellular genome was used, which resulted in the obtainment of C4 and 2D8GPR107-/-HaCaT monoclonal cell lines. CCK-8 assay and flow cytometry assay showed thatGPR107gene deletion resulted in G0G1phase block, significantly weakened proliferation ability and increased apoptosis level of HaCaT cells. Western blot found that the differentiation of HaCaT cells accelerated after knockdown ofGPR107. Additionally the results of the cell scratch assay indicated that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. The results showed that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. Conclusion HaCaT cell line withGPR107gene deletion is successfully constructed, GPR107 deletion blocks the G0G1phase of HaCaT cells, which inhibiting the proliferation of HaCaT cells and promoted apoptosis, and it was found that the differentiation and migration of HaCaT cells were enhanced after knocking downGPR107.

Objective:To translate Amyotrophic Lateral Sclerosis Impairment Multidomain Scale(AIMS)into Chinese,and to investi-gate its reliability and validity among Amyotrophic Lateral Sclerosis(ALS)patients.Methods:A total of 161 ALS patients who were registered in the registry system of Integrated Traditional Chinese and Western Medicine Treatment Center for Motor Neuron Disease in Hubei Provincial Hospital of Traditional Chinese Medicine from August 2023 to July 2024 were enrolled as subjects,and the Chinese version of AIMS was used for investigation after translation and revision.The Rasch model was used to assess the unidimensionality,in-ternal consistency,content validity,construct validity,and criterion-related validity of the questionnaire,and the classical test theory was used for item analysis,test-retest reliability analysis,and criterion-related validity analysis.Results:Unidimensionality testing showed that all three dimensions of AIMS have good unidimensionality.For all items except item 18,information-weighted fit statis-tic mean square(Infit MNSQ)and outlier-sensitive fit statistic mean square(Outfit MNSQ)ranged from 0.5 to 1.5,and point-measure correlation(PT-Measure)ranged from 0.63 to 0.87.The threshold between adjacent options of each item was>1.4 logit.Wright maps showed that item difficulties ranged from-2.24 to 2.08 logit,and the differential item functioning analysis showed that the absolute values of differences between patients across different char-acteristic subgroups were<1 logit.The overall scale and each di-mension had a reliability of>0.9,with an individual reliability of>0.7.The dimension of respiration had a separation index of 1.56,while the overall scale and the other dimensions had a separation in-dex of>2.The item analysis showed that the correlation coefficient between the items in each dimension and their respective dimen-sions ranged from 0.622 to 0.865(P<0.01).The overall scale and the dimensions of medulla oblongata,motor,and respiration had a test-retest reliability of 0.994,0.970,0.990,and 0.972,respectively(P<0.01).The criterion-related validity analysis showed that the correlation coefficients of total score and the score of each dimension between the Chinese version of AIMS and the revised version of AIMS were 0.863,0.829,0.876,and 0.755,respectively(P<0.01).Conclusion:The Chinese version of AIMS has a good degree of fit-ting and relatively high levels of reliability and validity,with moderate item difficulty,reasonable option settings,and high overall qual-ity,and it is suitable for the functional assessment of ALS patients in China.

Objective:To develop an automatic diagnostic model for temporomandibular joint osteoarthritis(TMJOA)imaging based on deep learning technology,and to assist clinical diagnosis and improve the efficiency and accuracy of TMJOA diagnosis.Methods:CBCT data of 220 patients were collected,and 2 052 sagittal images were exported.Regions of interest were delineated according to the imaging analysis criteria for temporomandibular joint disorders,and the images were classified into TMJOA-free,TMJOA-uncer-tain and TMJOA.The data were randomly divided into a training set and a validation set according to 8∶2 ratio,and the training set data were used to train a TMJOA detection model based on three lightweight YOLOV5 deep learning frameworks,and the models' performance was evaluated on the validation set.Results:The Yolov5N model demonstrated the best performance,achieving a de-tection accuracy,recall,and precision of 92.5％,90.1％and 85.7％on the validation set,respectively.Conclusion:The auto-matic detection model for TMJOA imaging developed in this study can effectively identify arthritic lesions.Artificial intelligence tools are expected to become a powerful auxiliary tool for the clinical diagnosis of TMJOA.

BACKGROUND:The abnormal aggregation of proteins and the loss of neurons are typical pathological changes observed in neurodegenerative diseases.Changes in the levels of O-linked N-acetylglucosamine glycosylation are closely related to relevant pathological changes and are promising potential therapeutic targets.OBJECTIVE:To review the role of O-linked N-acetylglucosamine glycosylation in neurodegenerative diseases and the current advancements in its clinical applications,aiming to provide new insights for the treatment of neurodegenerative disorders.METHODS:The first author conducted a literature search in the CNKI,VIP,WanFang Database,ChiCTR,Web of Science,PubMed,Cochrane Library,Clinical Trials and Alzforum,using the search terms"O-GIcNAcylation,Neurodegenerative diseases,Alzheimer's disease,Parkinson's disease,Amyotrophic lateral sclerosis,Huntington's disease,OGA inhibitors,Clinical trial"in Chinese and English.A total of 66 relevant articles were included for review.RESULTS AND CONCLUSION:O-linked N-acetylglucosamine glycosylation is involved in the development and functional regulation of neurons,and its levels gradually decrease as neuronal development matures.In neurodegenerative diseases,the pathological proteins involved are regulated by O-linked N-acetylglucosamine glycosylation.Most evidence suggests that using O-GIcNAcase inhibitors to enhance O-linked N-acetylglucosamine glycosylation levels can significantly alleviate related pathological changes,making it a potential therapeutic strategy for neurodegenerative diseases.Some O-GIcNAcase inhibitors have completed Phase I clinical trials,demonstrating good safety and tolerability.

Objective To elucidate the expression characteristics of the NUAK family SNF1-like kinase 2(NUAK2)in ovarian serous carcinoma and determine its regulatory roles in cell proliferation and apoptosis.Methods Fifty-seven patients with ovarian serous car-cinoma(observation group)and 57 patients with ovarian serous cystadenoma(control group)were selected from June 2019 to December 2022.The GEPIA database was used to assess the expression patterns of NUAK2 in ovarian serous carcinoma.Immunohistochemistry was used to detect the expression of NUAK2,proliferating cell nuclear antigen(PCNA),and B-cell lymphoma-associated X antigen(BAX).Western blotting was performed to evaluate the expression of NUAK2 in the human ovarian serous carcinoma cell lines(SKOV3)and the human ovarian surface epithelial cell lines(HOSEpiC).SKOV3 cells were transfected with the siRNA-NUAK2 plasmid to establish the si-NUAK2 group,or with the empty transfection vector(EV)or a blank solution(NC)to establish control groups.Western blotting was sub-sequently used to detect the expression of NUAK2,PCNA,and BAX.The CCK-8 method was used to assess cell proliferation activity,and flow cytometry was used to quantify apoptosis.Results The GEPIA database analysis showed a significant increasing trend in the expres-sion of NUAK2 in ovarian cancer(P<0.05).Immunohistochemical analysis revealed a significantly higher NUAK2 positive rate in ovarian serous carcinoma tissue than that in the control group.Moreover,significant differences in the expression of NUAK2 were observed across different lesion grades and maximum tumor diameters(P<0.05).A significant positive correlation was found between NUAK2 and PCNA(P<0.05),whereas a significant negative correlation was observed between NUAK2 and BAX(P<0.05)in ovarian serous carcinoma.In vitro cell culture experiments showed that the expression level of NUAK2 was significantly higher in SKOV3 cells than that in HOSEpiC cells(P<0.05).The si-NUAK2 group exhibited significantly lower cell proliferation activity and PCNA expression and significantly higher apoptosis rate and BAX expression than those in the EV and NC groups(P<0.05).Conclusion NUAK2 is overexpressed in ovarian serous carcinoma and promotes tumor progression by enhancing cell proliferation and suppressing apoptosis.

Objective:To analyze the awareness of HCV infection status and willingness for HCV-RNA testing among hepatitis C cases in four provinces in China and to provide a reference for adjusting HCV prevention and control strategies.Methods:From September 2021 to September 2022, a cross-sectional survey was conducted using stratified random cluster sampling in four provinces (Jiangsu, Henan, Hubei, and Yunnan) in China, with an estimated sample size of 6 468 participants. The questionnaire included sociodemographic information, HCV infection awareness, willingness for HCV-RNA testing, and history of high-risk behaviors from the survey participants. Logistic regression models were used to analyze the factors associated with HCV infection awareness and willingness for HCV-RNA testing among hepatitis C cases. Statistical analysis was performed using R 4.1.3 software.Results:A total of 10 563 hepatitis C cases were surveyed. The awareness rate of HCV infection was 86.74% (9 162/10 563), and the willingness rate for HCV-RNA testing was 85.21% (9 001/10 563). Multivariate logistic regression models analysis showed that the awareness rate of HCV infection was lower among individuals aged ≥60 years, urban residents (with New Rural Cooperative Medical Insurance ), those without a history of blood transfusion, those without a history of paid blood donation, those without a history of injection drug use, and those without a family member with hepatitis C case.The awareness rate was higher among individuals with high or technical secondary school education, college education or above, and those married with a spouse (all P<0.05). In terms of willingness for HCV-RNA testing, it was lower among females, individuals aged ≥60 years, and those without a history of blood transfusion, paid blood donation, or injection drug use. The willingness was higher among farmers or migrant workers, employees of enterprises or institutions, and those in other occupations (all P<0.05). Conclusions:There was room for improvement in the awareness proportion of HCV infection and willingness for HCV-RNA testing among hepatitis C cases in the four provinces of China. More convenient policies and measures should be provided to increase the awareness rate of HCV infection and the willingness to undergo HCV-RNA testing in this population.

Objective To evaluate the application value of fascial platform versus commercial platform in transumbilical single-port laparoscopic transabdominal preperitoneal repair(TAPP)for inguinal hernia.Methods One hundred patients with inguinal hernia admitted between January and December 2024 were prospectively enrolled and randomly assigned to either the fascial platform group(n=50)or the commercial platform group(n=50)by using a random number table.The following parameters were compared between the two groups:platform establishment time,operative time,intraoperative blood loss,incision suture time,operative difficulty score,camera manipulation difficulty score,postoperative 24-hour Visual Analogue Scale(VAS)score of pain,postoperative hospital stay,total hospitalization cost,comprehensive satisfaction score,and postoperative complications.Results The fascial platform group demonstrated significantly superior outcomes compared to the commercial platform group in platform establishment time[(3.4±1.0)min vs.(8.1±1.5)min,t=-18.389,P＜0.001],operative time[(67.5±7.9)min vs.(72.2±11.5)min,t=-2.380,P=0.019],intraoperative blood loss[(8.3±4.3)ml vs.(11.1±5.7)ml,t=-2.798,P=0.006],incision suture time[(7.0±1.4)min vs.(11.8±2.7)min,t=-11.212,P＜0.001],operative difficulty score(14.4±3.6 vs.23.1±4.1,t=-11.215,P＜0.001),camera manipulation difficulty score(11.4±2.6 vs.18.3±2.5,t=-13.370,P＜0.001),VAS score of postoperative 24-hour pain(2.4±1.0 vs.3.7±1.1,t=-6.139,P＜0.001),total hospitalization cost[(15.8±1.6)thousand yuan vs.(18.3±1.6)thousand yuan,t=-7.841,P＜0.001],and comprehensive satisfaction score(7.3±1.1 vs.6.3±1.0,t=5.271,P＜0.001).However,there were no statistically significant differences in postoperative hospital stay or postoperative complication between the two groups(P＞0.05).All the patients were followed up for 1-12 months postoperatively[(5.3±1.8)months vs.(5.7±2.0)months,t=-0.945,P=0.347],with no recurrence observed in either group.Conclusions Both fascial platform and commercial platform can be used for transumbilical single-port TAPP.The fascial platform offers advantages including shorter platform establishment time,reduced incision suture time,lower operative and camera manipulation difficulty,decreased cost,less postoperative pain,and higher patient satisfaction.