Objective This study reports on the obligatory parasitism of water mites Arrenurus sp. on Anopheles messeae at the Manzhouli Port, Inner Mongolia, China. Methods Duing July 2024, a survey on the mosquito diversity was conducted at the Manzhouli Port. Captured mosquitoes and their ectoparasites were identified to species level. Results A total of 1840 adult mosquitoes were collected, representing species from three genera: Culex(Cx. modestus, Cx. pipiens pallens), Aedes(Ae. dorsalis, Ae. flavidorsalis, Ae. flavescens), and Anopheles (An. messeae). Among all the mosqutioes specimens,3 out of 150 captured An. messeae were found to carry ectoparasitic mites, with number of 2,4,27 mites separately. Morphological and molecular identification reached the same result as water mites(Hydrachnidiae, Hydracrina). COI gene sequence showed 94% similarity with the closest species Arrenurus truncatellus. Conlusions Literature review suggests water mites are host-specific parasitism of mosquito species and herein with the first record of Arrenurus sp. parasiting on An. Messeae in the most high-latitude region globally.

ObjectiveColorectal cancer (CRC) is a highly heterogeneous malignancy, and traditional Chinese medicine (TCM) syndrome differentiation is widely used in its clinical management. However, the biological basis underlying different TCM syndromes remains insufficiently understood. This study aimed to investigate the proteomic differences between qi-blood deficiency syndrome (QBDS) and damp-heat syndrome (DHS) in CRC patients and to establish a “syndrome-differential protein-herbal component” network for exploring potential molecular mechanisms and therapeutic targets associated with syndrome-specific treatment strategies. MethodsTumor tissue samples were collected from 10 patients with pathologically confirmed CRC, including 5 patients diagnosed with QBDS and 5 with DHS according to standardized TCM syndrome differentiation criteria. Tandem mass tag (TMT)-based quantitative proteomics was employed to identify differentially expressed proteins (DEPs) between the two syndrome groups. Functional enrichment analyses, including Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were performed to characterize the biological functions and signaling pathways associated with the identified DEPs. Least absolute shrinkage and selection operator (LASSO) regression analysis was further applied to screen key syndrome-related proteins and construct a core protein prediction model. Subsequently, molecular docking was conducted to evaluate the binding affinities between representative herbal compounds, astragaloside IV and berberine, and the identified core proteins. Finally, cell counting kit-8 (CCK-8) assays were performed in HCT 116 and HT-29 colorectal cancer cell lines to validate the anti-proliferative activities of these compounds in vitro. ResultsA total of 44 DEPs were identified between QBDS and DHS tissues, including 21 upregulated and 23 downregulated proteins in the QBDS group relative to the DHS group. Functional enrichment analyses revealed that these proteins were primarily associated with antiviral immune responses, regulation of inflammatory signaling, NF‑κB signaling pathways, cholesterol metabolism, and other biological processes relevant to tumor progression and host immune regulation. LASSO regression analysis identified eight core proteins with potential syndrome-specific significance. Among them, ST7, GPCPD1, and PODXL were closely associated with QBDS, whereas FAT1, GRB7, KRT7, MAFF, and MMP8 were associated with DHS. Molecular docking demonstrated favorable interactions between astragaloside IV and QBDS-related proteins, with binding energies lower than -5 kcal/mol, indicating stable binding activity. Berberine exhibited even lower binding energies and stronger binding specificity toward DHS-related core proteins. In vitro experiments further confirmed that both compounds inhibited CRC cell proliferation, while berberine displayed significantly greater anti-proliferative efficacy. The lowest 48 h IC₅₀ value of berberine reached 15.21 mg/L in CRC cells, indicating a stronger growth-inhibitory effect than astragaloside IV. ConclusionCRC patients with QBDS and DHS exhibit distinct proteomic characteristics, suggesting that different TCM syndromes possess unique molecular signatures. The identified core proteins may serve as potential biomarkers for syndrome differentiation, while astragaloside IV and berberine may exert therapeutic effects through targeting syndrome-specific molecular networks. These findings provide preliminary proteomic evidence supporting syndrome-guided precision integrative medicine in CRC and offer new insights into the biological basis of TCM syndrome differentiation and individualized therapeutic strategies.

Objective To explore the influencing factors of different types of small intestinal bacterial overgrowth(SIBO).Methods A total of 539 patients who were hospitalized in the Department of Gastroenterology,the Sixth Medical Center of PLA General Hospital from June 2021 to December 2021 and who underwent methane-hydrogen breath test were retrospectively selected.Based on breath test results,patients were divided into SIBO-negative group(n=300)and SIBO-positive group(n=239).The clinical data were compared between two groups.According to the specific values of breath test results,SIBO-positive patients were further divided into hydrogen-producing bacterial overgrowth(hydrogen-positive,n=103),intestinal methanogen overgrowth(methanogen-positive,n=80),and simultaneous methanogen and hydrogen-producing bacterial overgrowth(double positive,n=56)groups.Multivariate logistic regression analysis was employed to identify influencing factors of different SIBO types.Additionally,SIBO-positive patients were categorized by age into<45 years(n=23),45-60 years(n=82),60-75 years(n=124),and≥75 years(n=10)to compare SIBO positivity rates across age groups.Results The patients in SIBO-positive and double positive groups were older and had a lower body mass index(BMI)than those in SIBO-negative group,with statistically significant differences(P<0.05).Compared with the patients in SIBO-negative group,those in hydrogen-positive group showed a higher proportion of history of coronary heart disease,those in methanogen-positive group were older,and higher proportion of statin use,with statistically significant differences(P<0.05).Multivariate logistic regression analysis revealed that,among different SIBO types,a history of coronary heart disease served as an independent risk factor for hydrogen-producing bacterial overgrowth(OR=2.728,95%CI 1.271-5.857,P=0.010).For methanogen overgrowth,increasing age was identified as an independent risk factor(OR=1.040,95%CI 1.009-1.063,P=0.010),while the application of statin played the role of an independent protective factor(OR=0.420,95%CI 0.236-0.754,P=0.003).As for the simultaneous overgrowth of methane-producing and hydrogen-producing bacteria,increased BMI was found to be an independent protective factor(OR=0.870,95%CI 0.786-0.964,P=0.008).In SIBO-positive group,it was found that for patients aged<45 years,both the methane-positive rate and the double-positive rate were significantly lower than the hydrogen positivity rate(P<0.05).Moreover,among patients aged 45-60 years,the double-positive rate was significantly lower than the hydrogen positivity rate(P<0.01).When it comes to the hydrogen-positive rate,it was significantly lower for patients aged 45-60 and 60-75 years compared with that of patients aged<45 years(P<0.05).In contrast,the methane-positive rate and the double-positive rate were significantly higher for patients aged 45-60 and 60-75 years than those of patients aged<45 years(P<0.01).Conclusion A history of coronary heart disease and increasing age are independent risk factors for intestinal hydrogen-producing bacterial overgrowth and methanogen overgrowth,respectively.The application of statins and increased BMI are independent protective factors for intestinal methanogen simultaneous overgrowth of methanogen and hydrogen-producing bacteria,respectively.

Objective This study aimed to investigate whether Trichinella spiralis Ts87 protein can facilitate immune evasion by degrading the mouse neutrophil extracellular traps(NETs).Methods First,we used bioinformatics tools to analyze the physicochemical properties of Ts87 and,through sequence homology alignment,confirmed its classification within the nuclease family.Molecular cloning and protein purification techniques were then employed to obtain high-purity Ts87 protein,and its nuclease activity was confirmed.Additionally,we compared the primary sequence of Ts87 with known active sites from human lysosomal DNase IIα and the structurally characterized Burkholderia thailandensis DNase II.This enabled us to predict potential active sites.Ts87 enzymatic mutants were generated using site-directed mutagenesis,and their functions were confirmed through enzymatic activity experiments.The extracellular nucleic acid content of mouse bone marrow neutrophils and the immunofluorescence staining of mouse NETs were assessed in vitro to verify whether the Ts87 protease mutants exhibited a decreased ability to degrade NETs.Finally,after immunizing mice with Ts87,calculated larvae burden and the degradation of NETs in vivo were observed by immunostaining the MPO and CitH3 expression in the intestinal NETs of the mice.Results High-purity Ts87 protein was successfully obtained and confirmed to possess nuclease activity,capable of degrading NETs in vitro.The Ts87 mutants exhibited reduced capacity in degrading NETs.In vivo experiments in mice demonstrated that the production of anti-Ts87 antibodies in immunized mice reduced the degradation of NETs,facilitating NET-mediated attack on the parasites and resulting in decreased larvae burden in the mice.Conclusions Trichinella spiralis can utilize Ts87 nuclease to evade capture by NETs,providing potential vaccine and drug targets for the prevention and treatment of trichinellosis.

Objective To establish a rapid,sensitive,and specific multiplex PCR detection method for the simultaneous detection of Cryptosporidium,Eimeria,and bovine parvovirus.Methods Specific primers targeting the SSU rRNA genes of Cryptosporidium and Eimeria,as well as the VP2 gene of bovine parvovirus were designed and the corresponding recombinant plasmid standards were constructed.To establish the multiplex PCR method,the reaction conditions were optimized using temperature gradient PCR and single-variable control methods.The sensitivity,specificity,reproducibility,and clinical application of the protocol were evaluated.Results The optimal annealing temperature was found to be 60.5℃,and the forward and reverse primer concentrations were determined to be 0.2 μmol/L for Eimeria,and 0.4 μmol/L for Cryptosporidium and bovine parvovirus.The assay demonstrated high sensitivity,with detection limits of 243,260,and 3 110 copies for the recombinant plasmid standards of Cryptosporidium,Eimeria,and bovine parvovirus,respectively.Specificity testing showed no cross-reactivity with ten common bovine pathogens,including Salmonella,bovine viral diarrhea virus,and bovine rotavirus.Consistent intra-and inter-batch results confirmed the strong reproducibility of the method.Clinical application to 81 diarrhea samples from various regions in the Ganzi Prefecture,Sichuan,revealed positivity rates of 18.52%(15/81)for Cryptosporidium,34.57%(28/81)for Eimeria,and 18.52%(15/81)forbovineparvovirus,withamixedinfectionrateof3.7%(3/81).Conclusions Themultiplex PCR method established in this study offers a reliable tool for differential diagnosis and epidemiological investigation of the three common diarrheal pathogens in yaks.

Hepatocellular carcinoma(HCC)expresses abundant glycolytic enzymes and displays comprehensive glucose metabolism reprogramming.Aldolase A(ALDOA)plays a prominent role in glycolysis;however,little is known about its role in HCC development.In the present study,we aim to explore how ALDOA is involved in HCC proliferation.HCC proliferation was markedly suppressed both in vitro and in vivo following ALDOA knockout,which is consistent with ALDOA overexpression encouraging HCC prolifera-tion.Mechanistically,ALDOA knockout partially limits the glycolytic flux in HCC cells.Meanwhile,ALDOA translocated to nuclei and directly interacted with c-Jun to facilitate its Thr93 phosphorylation by P21-activated protein kinase;ALDOA knockout markedly diminished c-Jun Thr93 phosphorylation and then dampened c-Jun transcription function.A crucial site Y364 mutation in ALDOA disrupted its interaction with c-Jun,and Y364S ALDOA expression failed to rescue cell proliferation in ALDOA deletion cells.In HCC patients,the expression level of ALDOA was correlated with the phosphorylation level of c-Jun(Thr93)and poor prognosis.Remarkably,hepatic ALDOA was significantly upregulated in the promotion and progression stages of diethylnitrosamine-induced HCC models,and the knockdown of Aldoa strikingly decreased HCC development in vivo.Our study demonstrated that ALDOA is a vital driver for HCC development by activating c-Jun-mediated oncogene transcription,opening additional avenues for anti-cancer therapies.

Objective To explore the correlation between visceral fat area(VFA)and triglyceride-glucose(TyG)index in people with normal body mass index(BMI)based on quantitative CT(QCT).Methods Totally 1 358 healthy adults with normal BMI who underwent QCT body composition measurement were retrospectively enrolled and divided into normal visceral fat group(normal group,VFA＜100 cm2,n=509),mild visceral fat accumulation group(mild group,100 cmn2≤VFA＜150 cm2,n=431)and severe visceral fat accumulation group(severe group,VFA≥150 cm2,n=418)according to VFA measured with QCT.Pearson and Spearman correlation analysis were performed to observe correlations of TyG index with general data and body composition in 20-40,41-60,61-90 years old men and women with normal BMI,respectively.Multiple linear regression analysis was performed to screen the independent impact factors of TyG index in normal BMI people.Results Low density lipoprotein cholesterol(LDL-C),total cholesterol(TC),glutamic-pyruvic transaminase(GPT),VFA and subcutaneous fat area(SFA)of men with normal BMI at different age stages were weakly-lowly positively correlated with TyG index(r=0.143-0.459,all P＜0.05),while high-density lipoprotein cholesterol(HDL-C)were weakly-lowly negatively correlated with TyG index(r=-0.410--0.222,all P＜0.05).LDL-C,VFA and SFA of women with normal BMI at different age stages were weakly-lowly positively correlated with TyG index(r=0.206-0.495,all P＜0.05).GPT,TC,VFA and liver fat content(LFC)were all independent risk factors,while HDL-C was independent protective factor for TyG index in people with normal BMI(all P＜0.05).Conclusion VFA was positively correlated with TyG index in people with normal BMI.Measurement of abdominal body composition based on QCT combined with lipid indexes had certain potential for predicting insulin resistance and relative metabolic diseases in abdominal obese people.

Objective To observe value of real-time functional MRI neurofeedback(rtfMRI-NF)for modulating brain activity changes in obese adults.Methods Twenty-two obese adults were prospectively recruited,3-week rtfMRI-NF training intervention was conducted.Barratt impulsivityness scale version 11(BIS-11),three-factor eating questionnaire(TFEQ)and food rating scales were scored before and after intervention,and whole-brain resting state fMRI(rs-fMRI)data were obtained.Clinical scale scores,rs-fMRI regional homogeneity(ReHo)values and amplitude of low frequency fluctuation(ALFF)values were compared before and after intervention.Then brain regions with differences of ReHo and ALFF values before and after intervention were explored,and correlations of values of rs-fMRI data differences and clinical scale score differences were analyzed.Results BIS-11 score,TFEQ emotional eating(TFEQ-EE)score and food rating scales scores decreased,while TFEQ cognitive restraint(TFEQ-CR)scores increased in obese adults after intervention(all P＜0.05).ReHo values in right inferior frontal gyrus,right anterior cingulate and left precuneus increased,while of left middle temporal gyrus decreased(all corrected P＜0.05).ALFF values of left precuneus increased,whereas of left middle occipital gyrus,right superior occipital gyrus,left calcarine fissure and surrounding cortex and left supramarginal and angular gyrus decreased(all corrected P＜0.05).ReHo difference in right anterior cingulate was negatively correlated with BIS-11 scores difference(r=-0.601,P＜0.05),and ALFF difference in left precuneus was negatively correlated with TFEQ-EE difference(r=-0.478,P＜0.05).Conclusion rtfMRI-NF intervention could correct disorder of cerebral functional areas in obese adults and change high-calorie food preference and poor dietary habits.

Diminished ovarian reserve (DOR) is a decrease in the number of available follicles in the ovaries and the quality of oocytes, which in turn affects follicle development, ovulation and sex hormone secretion, resulting in decreased fertility. Harmful chemical components in cigarette smoke, such as nicotine and benzopyrene, can interfere with the hypothalamic-pituitary-ovarian axis and impair ovarian reserve and function through apoptosis, autophagy, oxidative stress and DNA damage, which can directly affect ovarian granulosa cells and oocytes, causing oocyte development disorders, increased apoptosis of granulosa cells, destroy reproductive endocrine homeostasis, lead to DOR and affect female fertility. At present, the molecular mechanism of premature ovarian dysfunction caused by cigarette smoke is not fully understood. In this paper, the molecular mechanism of cigarette smoke exposure affecting ovarian tissue and mediating ovarian function was reviewed, aiming to provide theoretical reference for the study of cigarette smoke induced DOR and affecting ovarian function, and to provide pregnant guidance for infertile women in bad environment and bad living habits.

Objective:To correlate serum bone morphogenetic protein-2 (BMP-2) and cartilage oligomeric matrix protein (COMP) levels with joint function and systemic inflammatory response in patients with knee osteoarthritis (KOA) complicated by cartilage injury.Methods:This study is a retrospective analysis. It selected 258 patients with knee osteoarthritis who were treated at Yantai Yuhuangding Hospital from January 2022 to December 2023. All patients underwent arthroscopic examination. Based on the presence of cartilage injury, 118 patients with cartilage injury were included in the Group A, while 140 patients without cartilage injury were included in the Group B. Joint function (WOMAC Osteoarthritis Index, Lysholm Knee Function Score), serum levels of BMP-2 and COMP, and systemic inflammatory responses [levels of interleukin (IL)-1, IL-6, and tumor necrosis factor-alpha (TNF-α)] were compared between the two groups. Receiver Operating Characteristic (ROC) curves were drawn to analyze the diagnostic efficacy of serum BMP-2 and COMP levels for KOA complicated by cartilage injury. Pearson correlation analysis was used to assess the correlation between the aforementioned serum markers and the patients' WOMAC Osteoarthritis Index, Lysholm Knee Function Score, as well as serum levels of IL-1, IL-6, and TNF-α.Results:Group A had significantly lower serum BMP-2 levels and Lysholm Knee Function Scores compared with Group B [(9.18 ± 1.65) ng/L vs. (11.43 ± 2.51) ng/L, t = 8.34, P < 0.001; (80.06 ± 8.34) vs. (86.94 ± 7.21), t = 7.11, P < 0.001]. Group A exhibited higher serum levels of COMP, IL-1, IL-6, and TNF-α, as well as a higher WOMAC Osteoarthritis Index, with all differences being statistically significant [(6.88 ± 2.34) μg/L vs. (5.04 ± 1.01) μg/L, t = -8.34, P < 0.001; (43.18 ± 7.81) ng/L vs. (35.96 ± 5.02) ng/L, t = -8.96, P < 0.001; (34.03 ± 6.68) ng/L vs. (28.75 ± 5.61) ng/L, t = -6.90, P < 0.001; (13.17 ± 1.89) ng/L vs. (11.24 ± 1.01) ng/L, t = -10.44, P < 0.001; (137.18 ± 18.95) vs. (121.14 ± 13.58), t = -7.90, P < 0.001]. ROC curve analysis indicated that the area under the curve values for serum BMP-2 and COMP levels in diagnosing KOA complicated by cartilage injury were 0.773 and 0.811, respectively. Pearson correlation analysis revealed a negative correlation between serum BMP-2 levels and the WOMAC Osteoarthritis Index, as well as serum levels of IL-1, IL-6, and TNF-α ( r = -0.52, -0.42, -0.40, -0.57, all P < 0.05). Additionally, serum BMP-2 levels showed a positive correlation with the Lysholm Knee Function Score ( r = 0.51, P < 0.05). Serum COMP levels and the WOMAC Osteoarthritis Index were positively correlated with serum IL-1, IL-6, and TNF-α levels ( r = 0.48, 0.45, 0.37, 0.54, all P < 0.05) and were negatively correlated with the Lysholm Knee Function Score ( r = -0.51, P < 0.05). Conclusions:Serum BMP-2 and COMP levels have certain diagnostic value in patients with KOA complicated by cartilage injury. Moreover, these indicators are correlated with joint function and systemic inflammatory responses. Therefore, they hold promise as potential indicators for assessing disease progression in patients with KOA.