ObjectiveHepatocellular carcinoma (HCC) represents 90% of all primary liver cancers. The main risk factors associated with HCC include viral hepatitis (B and/or C), alcohol abuse, and metabolic dysfunction-associated steatotic liver disease (MASLD), which progressively advance to liver fibrosis, cirrhosis, and ultimately evolve into HCC. Surgical resection represents the most effective treatment for HCC, while recent advances in immunotherapy, including immune checkpoint inhibitors and adoptive cell therapies, have provided improved treatment prospects for patients with unresectable HCC. However, the complex metabolic heterogeneity of HCC limits the therapeutic efficacy. Metabolic intermediates acyl-CoA not only provide energy and substrates for numerous biochemical reactions but also serve as donors for protein lysine acylation, a major class of post-translational modification (PTM). Therefore, a deeper understanding of the molecular mechanisms underlying protein lysine acylation and hepatocarcinogenesis is urgently needed. MethodsThe levels of protein lysine acylation and silence information regulator 5 (SIRT5) expression levels in clinical HCC samples were analyzed by Western blot. Quantitative malonylome and succinylome of HCC samples were analyzed by antibody-based affinity enrichment coupled with tandem mass spectrometry. The proliferation of HCC cells was analyzed with Cell Counting Kit-8 (CCK-8) assays, the apoptosis was quantified by Annexin V-FITC/propidium iodide (PI) staining coupled with flow cytometry, and the ability of cells to migrate was assayed by Transwell assays. The enzymatic activity of glutathione S-transferase Mu 1 (GSTM1) was quantified. Transgenic mice with hepatic overexpression of SIRT5 were constructed using CRISPR-Cas9, and primary hepatocarcinogenesis was induced by administration of diethylnitrosamine. ResultsWestern blot analysis indicated that the expression level of SIRT5 was elevated in clinical samples from HCC patients, and the levels of lysine malonylation, glutarylation, and succinylation were significantly reduced in HCC tissues. Knockout of SIRT5 in MHCC-97H and MHCC-97L hepatoma cells suppressed cell proliferation, and increased the percentage of apoptotic cells significantly. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the differentially malonylome and succinylome of HCC samples revealed significant enrichment in two major classes of biological processes: core energy metabolism (e.g., glycolysis/gluconeogenesis, tricarboxylic acid metabolic process, fatty acid beta oxidation) and detoxification and oxidative stress response (e.g., response to toxic substance, chemical carcinogenesis, reactive oxygen species (ROS)). SIRT5 removes malonylation from lysine residues in GSTM1 and restores its detoxification activity, which is crucial for the survival of hepatocytes under stressed conditions. More importantly, in vivo experiment indicated that hepatic-specific overexpression of SIRT5 in mice accelerated diethylnitrosamine-induced liver fibrosis and hepatocarcinogenesis, indicating the critical role of SIRT5 in HCC progression. ConclusionThis study highlights the previously unrecognized SIRT5-GSTM1 axis as a key regulator in hepatocarcinogenesis, and suggests a potential target for the treatment of patients with HCC.

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.

Objective:To investigate the pharmacological mechanism of Compound Xuanju Capsule in the treatment of erectile dysfunction(ED)by using network pharmacology and molecular docking technology.Methods:The active ingredients and targets of Compound Xuanju Capsule were screened using Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform(TCMSP).TTD,OMIM,DrugBank and GeneCards databases were used to obtain genes related to ED,and the union of the results was taken as the disease genes of ED.The common target of drug and disease was taken as the potential target of Compound Xuanju Capsule in ED,and the drug-disease interaction network was constructed by using Cytoscape software.The protein-protein interaction(PPI)network was constructed by using String database,which was then imported into Cytoscape to identify the key target.Based on the drug-disease intersection genes,GO and KEGG enrichment analyses were performed to predict the relevant signaling pathways and molecular mechanisms of Compound Xuanju Capsule for the treatment of ED.Autodock software was used to perform molecular docking between the active ingredients and the core targets.Results:Forty chemical components of Compound Xuanju Capsule were screened,and 239 predicted targets were obtained.A total of 1 907 ED-related genes were screened,and 97 common targets were identified between Compound Xuanju Capsule and ED,among which the core targets included EGFR,ESR1,HIF1A,PTGS2,and STAT3.The signaling pathways obtained by KEGG enrichment analysis included calcium signaling pathway,HIF-1 signaling pathway,PI3K-Akt signaling pathway,cGMP-PKG signaling pathway,relaxin signaling pathway,Serotonergic synapse signaling pathway.The molecular docking results showed that there were molecular binding sites between the key active ingredients and the core targets with strong binding activity.Conclusion:Compound Xuanju Capsule may treat ED through multi-target pathways such as anti-inflamnmato-ry and improving cellular oxidative stress.

Objective:To explore serological detection and blood transfusion strategies of mimicking antibodies,so as to provide appropriate transfusion strategies.Methods:Detailed serological tests,including ABO blood group,Rh typing,antibody specificity,etc,were performed on two patients with autoimmune hemolytic anemia(AIHA).Meanwhile,the references about blood transfusion from mimicking antibody patients published from 1977 to 2024 in China and abroad were retrospectively summarized and analyzed.Results:The patient 1 blood type was AB,CCDee and the antibody is mimicking anti-e,transfusion the e-negative red blood cells(RBCs)was effective.After two transfusions of e-RBCs,hemoglobin levels significantly increased from 48 g/L to 91 g/L,with complete resolution of hemolytic symptoms.The patient 2 blood type was O,CcDee,and the antibody was mimicking anti-c,the patient was diagnosed with AIHA and treated with hormone.No blood products were transfused during hospitalization,and his hemolysis was relieved.Conclusion:Strictly grasping the indication of blood transfusion,blood transfusion should not be performed in the unnecessary conditions,and the corresponding antigen-negative RBC should be screened for transfusion in the necessay conditions.

Objective To establish a nomogram prediction model for the risk of sarcopenia among elderly patients with type 2 diabetes mellitus(T2DM)by LASSO-Logistic regression.Methods From January 2021 to December 2023,2986 elderly T2DM patients who were admitted to The Fifth People's Hospital of Chongqing were enrolled.Based on the 2019 criteria of the Asian Sarcopenia Working Group,they were divided into a T2DM combined with sarcopenia(Sar,n=867)group and simple T2DM group(n=2119).All patients were randomly divided into a model group(n=1990)and a validation group(n=996)in a 2∶1 ratio.Another 528 patients with T2DM admitted to our hospital from January to October 2024 were selected as the external validation group.LASSO regression was used to screen variables.Logistic regression was used to analyze the influencing factors of sarcopenia.R language was used to establish a nomogram model.Receiver operating characteristic(ROC)curve and Hosmer-Lemeshow curve were used to validate the prediction model.Results The age,proportions of drinking,hypertension,hyperlipidemia,DM complications,DM duration,fasting plasma glucose(FPG),2-hour postprandial plasma glucose,hemoglobin A1c(HbA1c),triglycerides,total cholesterol,and low-density lipoprotein cholesterol in the Sar group were higher than those in the T2DM group(P<0.05).The proportion of male individuals,body mass index,and high-density lipoprotein cholesterol in the Sar group were lower than those in the T2DM group(P<0.05).LASSO regression identified 8 factors,including gender,age,drinking,hypertension,DM complications,DM duration,FPG,and HbA1c,as predictive variables for T2DM with sarcopenia.Logistic regression analysis showed that gender,age,DM duration,HbA1c,and DM complications were influencing factors for sarcopenia in elderly T2DM patients.The nomogram prediction model showed that the highest scores corresponding to DM complications,DM duration,HbA1c,age,and gender were approximately 100,70,52,43,and 30 points,respectively,with a maximum total score of approximately 295.The area under the ROC curve of the model group,validation group,and external validation group were 0.914,0.822,and 0.777,respectively.The Hosmer-Lemeshow calibration curve showed that the maximum offsets for the model group,validation group,and external validation group were 0.031,0.049,and 0.056,respectively(P=0.929,0.802,and 0.782,respectively).Conclusions Elderly T2DM patients are prone to developing sarcopenia.A nomogram model based on LASSO-Logistic analysis,which includes DM complications,DM duration,HbA1c,age,and gender,can effectively predict the risk of sarcopenia among elderly T2DM patients.

This study aims to investigate the effect of Chaijin Jieyu Anshen Formula on the neuroinflammation of rats with insomnia complicated with depression through the regulation of triggering receptor expressed on myeloid cells 2(TREM2)/complement protein C1q signaling pathway. Rats were randomly divided into a normal group, a model group, a positive drug group, as well as a high, medium, and low-dose groups of Chaijin Jieyu Anshen Formula, with 10 rats in each group. Except for the normal group, the other groups were injected with p-chlorophenylalanine and exposed to chronic unpredictable mild stress to establish the rat model of insomnia complicated with depression. The sucrose preference experiment, open field experiment, and water maze test were performed to evaluate the depression in rats. Enzyme-linked immunosorbent assay was employed to detect serum 5-hydroxytryptamine(5-HT), dopamine(DA), and norepinephrine(NE) levels. Hematoxylin and eosin staining and Nissl staining were used to observe the damage in nucleus accumbens neurons. Western blot and immunofluorescence were performed to detect TREM2, C1q, postsynaptic density 95(PSD-95), and synaptophysin 1(SYN1) expressions in rat nucleus accumbens, respectively. Golgi-Cox staining was utilized to observe the synaptic spine density of nucleus accumbens neurons. The results show that, compared with the model group, Chaijin Jieyu Anshen Formula can significantly increase the sucrose preference as well as the distance and number of voluntary activities, shorten the immobility time in forced swimming test and the successful incubation period of positioning navigation, and prolong the stay time of space exploration in the target quadrant test. The serum 5-HT, DA, and NE contents in the model group are significantly lower than those in the normal group, with the above contents significantly increased after the intervention of Chaijin Jieyu Anshen Formula. In addition, Chaijin Jieyu Anshen Formula can alleviate pathological damages such as swelling and loose arrangement of tissue cells in the nucleus accumbens, while increasing the Nissl body numbers. Chaijin Jieyu Anshen Formula can improve synaptic damage in the nucleus accumbens and increase the synaptic spine density. Compared to the normal group, the expression of C1q protein was significantly higher in the model group, while the expression of TREM2 protein was significantly lower. Compared to the model group, the intervention with Chaijin Jieyu Anshen Formula significantly downregulated the expression of C1q protein and significantly upregulated the expression of TREM2. Compared with the model group, the PSD-95 and SYN1 fluorescence intensity is significantly increased in the groups receiving different doses of Chaijin Jieyu Anshen Formula. In summary, Chaijin Jieyu Anshen Formula can reduce the C1q protein expression, relieve the TREM2 inhibition, and promote the synapse-related proteins PSD-95 and SNY1 expression. Chaijin Jieyu Anshen Formula improves synaptic injury of the nucleus accumbens neurons, thereby treating insomnia complicated with depression.
Animals ; Male ; Rats ; Nucleus Accumbens/metabolism* ; Drugs, Chinese Herbal/administration & dosage* ; Depression/complications* ; Membrane Glycoproteins/genetics* ; Rats, Sprague-Dawley ; Sleep Initiation and Maintenance Disorders/complications* ; Neurons/metabolism* ; Receptors, Immunologic/genetics* ; Signal Transduction/drug effects* ; Synapses/metabolism*

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.

Objective To establish a nomogram prediction model for the risk of sarcopenia among elderly patients with type 2 diabetes mellitus(T2DM)by LASSO-Logistic regression.Methods From January 2021 to December 2023,2986 elderly T2DM patients who were admitted to The Fifth People's Hospital of Chongqing were enrolled.Based on the 2019 criteria of the Asian Sarcopenia Working Group,they were divided into a T2DM combined with sarcopenia(Sar,n=867)group and simple T2DM group(n=2119).All patients were randomly divided into a model group(n=1990)and a validation group(n=996)in a 2∶1 ratio.Another 528 patients with T2DM admitted to our hospital from January to October 2024 were selected as the external validation group.LASSO regression was used to screen variables.Logistic regression was used to analyze the influencing factors of sarcopenia.R language was used to establish a nomogram model.Receiver operating characteristic(ROC)curve and Hosmer-Lemeshow curve were used to validate the prediction model.Results The age,proportions of drinking,hypertension,hyperlipidemia,DM complications,DM duration,fasting plasma glucose(FPG),2-hour postprandial plasma glucose,hemoglobin A1c(HbA1c),triglycerides,total cholesterol,and low-density lipoprotein cholesterol in the Sar group were higher than those in the T2DM group(P<0.05).The proportion of male individuals,body mass index,and high-density lipoprotein cholesterol in the Sar group were lower than those in the T2DM group(P<0.05).LASSO regression identified 8 factors,including gender,age,drinking,hypertension,DM complications,DM duration,FPG,and HbA1c,as predictive variables for T2DM with sarcopenia.Logistic regression analysis showed that gender,age,DM duration,HbA1c,and DM complications were influencing factors for sarcopenia in elderly T2DM patients.The nomogram prediction model showed that the highest scores corresponding to DM complications,DM duration,HbA1c,age,and gender were approximately 100,70,52,43,and 30 points,respectively,with a maximum total score of approximately 295.The area under the ROC curve of the model group,validation group,and external validation group were 0.914,0.822,and 0.777,respectively.The Hosmer-Lemeshow calibration curve showed that the maximum offsets for the model group,validation group,and external validation group were 0.031,0.049,and 0.056,respectively(P=0.929,0.802,and 0.782,respectively).Conclusions Elderly T2DM patients are prone to developing sarcopenia.A nomogram model based on LASSO-Logistic analysis,which includes DM complications,DM duration,HbA1c,age,and gender,can effectively predict the risk of sarcopenia among elderly T2DM patients.

Radiation-induced thrombocytopenia (RIT) faces a perplexing challenge in the clinical treatment of cancer patients, and current therapeutic approaches are inadequate in the clinical settings. In this research, oxymatrine, a new molecule capable of healing RIT was screened out, and the underlying regulatory mechanism associated with magakaryocyte (MK) differentiation and thrombopoiesis was demonstrated. The capacity of oxymatrine to induce MK differentiation was verified in K-562 and Meg-01 cells in vitro. The ability to induce thrombopoiesis was subsequently demonstrated in Tg (cd41:enhanced green fluorescent protein (eGFP)) zebrafish and RIT model mice. In addition, we carried out network pharmacological prediction, drug affinity responsive target stability assay (DARTS) and cellular thermal shift assay (CETSA) analyses to explore the potential targets of oxymatrine. Moreover, the pathway underlying the effects of oxymatrine was determined by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, Western blot (WB), and immunofluorescence. Oxymatrine markedly promoted MK differentiation and maturation in vitro. Moreover, oxymatrine induced thrombopoiesis in Tg (cd41:eGFP) zebrafish and accelerated thrombopoiesis and platelet function recovery in RIT model mice. Mechanistically, oxymatrine directly binds to toll-like receptor 2 (TLR2) and further regulates the downstream pathway stimulator of interferon genes (STING)/nuclear factor-kappaB (NF-κB), which can be blocked by C29 and C-176, which are specific inhibitors of TLR2 and STING, respectively. Taken together, we demonstrated that oxymatrine, a novel TLR2 agonist, plays a critical role in accelerating MK differentiation and thrombopoiesis via the STING/NF-κB axis, suggesting that oxymatrine is a promising candidate for RIT therapy.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.