ObjectiveTo obtain the gene sequences of major histocompatibility complex (MHC ) Ⅱgenes of Wuzhishan pigs, analyze their genetic information, and explore the biological functions of their MHC system. MethodsSpleen samples were collected from 3 adult male Wuzhishan pigs. Primers were designed according to MHCⅡ gene sequences, and the coding sequences of Wuzhishan pig MHCⅡ genes were amplified by RT-PCR. Sanger sequencing was performed to determine the full-length sequences. Bioinformatics tools were employed to analyze the physicochemical properties, phylogenetic relationships, conserved motifs, structural domains, chromosomal localization, and syntenic relationships of these genes. ResultsEight MHCⅡ genes were identified in Wuzhishan pigs, designated as SLA-DRA, SLA-DQA, SLA-DQB, SLA-DRB, SLA-DOB, SLA-DMB, SLA-DMA and SLA-DOA. The full-length sequences of these genes were determined by Sanger sequencing and subsequently deposited in GenBank under accession numbers PQ182796, PQ182797, PQ182798, PQ182799, PQ182800, PQ182801, PQ182802, and PQ164779. Phylogenetic analysis showed that the six MHCⅡ genes of Wuzhishan pigs clustered separately from their counterparts in Duroc, Meishan, Large White, and Bama pigs, indicating distinct evolutionary trajectories. Bioinformatics analysis demonstrated that most MHC Ⅱ proteins were hydrophobic, with molecular weights ranging from 27 700 to 30 000 Da. Genes within the same subregion shared conserved motifs. Specifically, four MHCⅡ proteins encoded by SLA-DQB, SLA-DRB, SLA-DOB, and SLA-DMB contained the MHCⅡβ conserved domain, while those encoded by the genes SLA-DRA, SLA-DQA, SLA-DMA, and SLA-DOA contained the MHCⅡα conserved domain. The eight MHCⅡ genes were scattered along the long arm of chromosome 7 in the Wuzhishan pigs, exhibiting syntenic relationships with three human genes and five Duroc pig genes. ConclusionThe MHCⅡ genes of Wuzhishan pigs may possess a unique evolutionary origin.

ObjectiveTo obtain the gene sequences of major histocompatibility complex (MHC ) Ⅱgenes of Wuzhishan pigs, analyze their genetic information, and explore the biological functions of their MHC system. MethodsSpleen samples were collected from 3 adult male Wuzhishan pigs. Primers were designed according to MHCⅡ gene sequences, and the coding sequences of Wuzhishan pig MHCⅡ genes were amplified by RT-PCR. Sanger sequencing was performed to determine the full-length sequences. Bioinformatics tools were employed to analyze the physicochemical properties, phylogenetic relationships, conserved motifs, structural domains, chromosomal localization, and syntenic relationships of these genes. ResultsEight MHCⅡ genes were identified in Wuzhishan pigs, designated as SLA-DRA, SLA-DQA, SLA-DQB, SLA-DRB, SLA-DOB, SLA-DMB, SLA-DMA and SLA-DOA. The full-length sequences of these genes were determined by Sanger sequencing and subsequently deposited in GenBank under accession numbers PQ182796, PQ182797, PQ182798, PQ182799, PQ182800, PQ182801, PQ182802, and PQ164779. Phylogenetic analysis showed that the six MHCⅡ genes of Wuzhishan pigs clustered separately from their counterparts in Duroc, Meishan, Large White, and Bama pigs, indicating distinct evolutionary trajectories. Bioinformatics analysis demonstrated that most MHC Ⅱ proteins were hydrophobic, with molecular weights ranging from 27 700 to 30 000 Da. Genes within the same subregion shared conserved motifs. Specifically, four MHCⅡ proteins encoded by SLA-DQB, SLA-DRB, SLA-DOB, and SLA-DMB contained the MHCⅡβ conserved domain, while those encoded by the genes SLA-DRA, SLA-DQA, SLA-DMA, and SLA-DOA contained the MHCⅡα conserved domain. The eight MHCⅡ genes were scattered along the long arm of chromosome 7 in the Wuzhishan pigs, exhibiting syntenic relationships with three human genes and five Duroc pig genes. ConclusionThe MHCⅡ genes of Wuzhishan pigs may possess a unique evolutionary origin.

Objective:To retrospectively analyze the results of auditory examination,vestibular function examination and laboratory examination of 63 patients diagnosed as vestibular neuritis.Methods:A total of 63 patients diagnosed with vestibular neuritis hospitalized in the Department of Otolaryngology, Head and Neck Surgery of the Third Affiliated Hospital of Sun Yat-sen University, from October 2012 to December 2022 were recruited. All patients met the diagnostic criteria for the 2022 Bárány association vestibular neuritis. Clinical data and the results of pure tone audiometry, electrocochleogram, video electronystagmogram, caloric test, cervical vestibuloevoked myogenic potential（cVEMP）, ocular vestibuloevoked myogenic potential（oVEMP）, video head impulse test（vHIT） was collected.A total of 63 age-and sex-matched healthy subjects in the physical examination center were randomly selected as the control group. The differences of blood indexs and lipid metabolism indexes between the two groups were compared. Results:In patients with vestibular neuritis, 50 out of 63 patients presented normal threshold in pure tone audiometry, 8 out of 63 patients had bilateral high-frequency sensorineural hearing loss and 5 out of 63 patients had unilateral mild high-frequency sensorineural hearing loss, 56 out of 63 cases completed the electrocochleogram, of which 3 cases had a binaural-SP/AP amplitude ratio≥0.4, 5 cases had monaural amplitude ratio ≥0.4. Fifty-five out of 63 patients completed the caloric test with CP values greater than 30% in all. The ratio of patients completed cVEMP, oVEMP and vHIT were 46 cases, 22 cases and 30 cases, respectively. 17 out of 63 cases completed all the four vestibular function tests. According to these tests, 49 patients could determine the extent of injury，including 27 cases with unilateral superior vestibular nerve injury, 21 cases with unilateral superior and inferior vestibular nerve injury and 1 case with unilateral inferior vestibular nerve injury. There were significant differences in neutrophil value（P<0.001）, lymphocyte value（P<0.005）, neutrophil/lymphocyte ratio（P<0.001） and apolipoprotein A1（P<0.001） between patient group and control group. Inflammatory markers were risk factors for patients with vestibular neuritis. The OR values of neutrophil value and blood neutrophil/lymphocyte ratio were 1.81（1.38-2.37, P<0.001） and 2.11（1.41-3.16, P<0.001）, respectively. Apolipoprotein A1 was a protective factor for patients with vestibular neuritis, and the OR value was 0.004（0.001-0.042, P<0.001）. Conclusion:Electrocochleogram could be used in vestibular neuritis patients with normal pure tone threshold to test whether there is hidden hearing loss in these patients. Neutrophil value, lymphocyte value, neutrophil/lymphocyte ratio and apolipoprotein A1 were correlated with vestibular neuritis. The Neutrophil value and neutrophil/lymphocyte ratio were risk factors for morbidity.
Humans ; Vestibular Neuronitis/physiopathology* ; Retrospective Studies ; Female ; Male ; Audiometry, Pure-Tone ; Hearing Loss, Sensorineural/physiopathology* ; Middle Aged ; Adult ; Vestibular Function Tests ; Vestibular Evoked Myogenic Potentials ; Aged

OBJECTIVES: To investigate the neural mechanisms of rhythmic activity in the hippocampal CA1 region and orbitofrontal cortex (OFC) during a spatial goal-directed task. METHODS: Four long-Evans rats were trained to perform a spatial goal-directed task in a land-based water maze (Cheese-board maze). The task was divided into 5 periods: Pre-test, Pre-sleep, Learning, Post-sleep, and Post-test. During the Learning phase, the task was split into two goal navigation and two reward acquisition processes with a total of 8 learning stages. Local field potentials (LFP) from the CA1 and the OFC were recorded, and power spectral density analysis was performed on Theta (6-12 Hz), Beta (15-30 Hz), Low gamma (30-60 Hz), and High gamma (60-90 Hz) bands. Coherence, phase-locking value (PLV), and phase-amplitude cross coupling (PAC) were used to assess the interactions between the CA1 and the OFC during learning and memory. RESULTS: During the task training, the rats showed consistent rhythms of OFC neural activity across the task states (P>0.05) while exhibiting significant changes in Beta and High gamma rhythms in the CA1 region (P<0.05). Coherence and PLV between the CA1 and the OFC were higher during goal navigation, especially in the stable learning phase (Stage 8 vs Stage 1, P<0.01). The rats showed stronger cross-frequency coupling between CA1-Theta and OFC-Low gamma in the Post-test phase than in the Pre-test phase (P<0.05). CONCLUSIONS Learning and memory consolidation in goal-directed tasks involve synchronized activity between the CA1 region and the OFC, and cross-frequency coupling plays a key role in maintaining short-term memory of reward locations in rats.
Animals ; Rats ; Rats, Long-Evans ; CA1 Region, Hippocampal/physiology* ; Memory Consolidation/physiology* ; Prefrontal Cortex/physiology* ; Maze Learning/physiology* ; Goals ; Male ; Memory/physiology* ; Learning/physiology*

Objective To investigate the risk factors of mild cognitive impairment(MCI)in elderly pa-tients with hypertension.Methods 270 elderly hypertensive patients treated in the geriatrics department and cardiovascular department of our hospital from September 2023 to March 2024 were analyzed retrospectively.They were divided into hypertension group with MCI(n=142)and hypertension group without MCI(n=128)according to the score of the simple mental state examination(MMSE).The clinical data and laboratory indicators of the two groups were collected and compared.The risk factors of MCI in elderly hospitalized pa-tients with hypertension were analyzed by stepwise logistic regression,and the diagnostic efficiency was ana-lyzed by receiver operating characteristic(ROC)curve.Results The prevalence rate of MCI in elderly pa-tients with hypertension was 52.6％.The monofactor analysis showed that the two groups had different hy-pertension years,hypertension grade,education level,combined diabetes,fasting blood glucose(FBG),triglyc-eride(TG),high density lipoprotein-cholesterol(HDL-C),HDL-C/low density lipoprotein-cholesterol(LDL-C),neutrophil(NEU),lymphocyte(LYM),glycated hemoglobin(HbA1c),arteriosclerosis index(AI),atherogenic index of plasma(AIP)and lipid composite index(LCI),the differences were statistically signifi-cant(P＜0.05).Multivariate logistic regression results showed that hypertension grade(grade 3),diabetes melli-tus,NEU,HbA1c,LYM and AIP were the influencing factors for MCI in elderly hospitalized patients with hyperten-sion(P＜0.05).The area under the curve(AUC)of FBG,TG,HbA1c and AIP in the diagnosis of hypertension com-bined with MCI were 0.801,0.743,0.889 and 0.804,respectively.Conclusion Early management and intervention can be carried out on the influencing factors of MCI in elderly patients with hypertension.

Objective To investigate the nutritional quality characteristics of vitamin D3-fortified yogurt and explore its improving effect on vitamin D metabolism in the body under simulated weightlessness,thereby providing a theoretical basis for the development of functional foods.Methods Using reconstituted milk as the matrix and Vitamin D3(VD3)microcapsule powder as the fortifier,VD3-fortified yogurt was prepared.A systematic study was conducted to investigate the effects of different gradients(1.25 μg/100 mL,2.50 μg/100 mL,3.75 μg/100 mL,5.00 μg/100 mL,6.25 μg/100 mL)of VD3 microcapsule addition on its quality characteristics(titratable acidity,solid content,water-holding capacity,syneresis).In vivo assessments were conducted using a Sprague-Dawley(SD)rat tail-suspension model to simulate weightlessness.Levels in serum 25(OH)D3,1,25-(OH)2D3,calcium(Ca),and phosphorus(P)were detected using the enzyme-linked immunosorbent assay(ELISA)to evaluate its metabolic capacity.Results During fermentation(3 h),titratable acidity of VD?-fortified yogurt initially increased,then decreased,and eventually stabilized with rising microcapsule dosage,while total solid content remained consistent.WHC exhibited an initial increase followed by a decline,whereas syneresis showed an inverse trend.At an optimal dosage of 3.75 μg/100 mL,the yogurt displayed a dense and uniform network structure,characterized by non-Newtonian fluid behavior with shear-thinning properties.This formulation demonstrated robust structural stability under high-frequency mechanical stress,alongside desirable textural,flavor,and sensory attributes.Animal experiments revealed that the serum concentrations of 25(OH)D3,1,25-(OH)2D3,calcium,and phosphorus in the vitamin D?-fortified yogurt intervention group were significantly higher than those in the tail-suspended control group(P<0.05).Conclusion VD? microencapsulation technology effectively preserves and enhances the nutritional quality characteristics of yogurt and mitigates vitamin D metabolic dysregulation under simulated weightlessness.

Pancreatic cancer is characterized by an insidious onset,high invasiveness,and poor response to treatment.Radical resection remains the only potentially curative approach currently available.However,approximately 80%of patients are already in the locally advanced or metastatic stage at initial diagnosis and have missed the opportunity for radical surgery.In recent years,with the development of novel drugs and updates to chemotherapy regimens,significant progress has been made in improving the efficacy of conversion therapy.Conversion therapy aims to transform initially unresectable tumors,such as locally advanced or metastatic pancreatic cancer,into a resectable state through systemic therapies(including chemotherapy,targeted therapy,immunotherapy,etc.)combined with localized treatments(such as radiotherapy,interventional therapy,etc.).This approach reduces tumor volume and eliminates micrometastases,thereby improving surgical resection rates and patient survival outcomes.However,considerable controversy remains regarding surgical treatment after conversion therapy for pancreatic cancer.This article provides an overview of conversion therapy regimens and treatment cycles,methods for evaluating therapeutic efficacy post-conversion therapy,the resection rates,and treatment outcomes of surgery following conversion therapy.The key technical points of post-conversion therapy surgery,including R0 resection,venous management,arterial resection and reconstruction,and the management of periarterial divestment,are highlighted.The learning curve for arterial resection and reconstruction is also discussed.Additionally,the potential applications of minimally invasive techniques(such as laparoscopy and robotics)are highlighted.The integration of biomarkers,imaging,and artificial intelligence holds promise for optimizing individualized decision-making.Multidisciplinary collaboration,the comprehensive competence of the surgical team,and the mastery and refinement of surgical skills are also critical factors that influence resectability and therapeutic outcomes after conversion therapy.

In recent years， the research method of Mendelian randomization based on genome-wide association studies has been widely used for etiological exploration in the medical field， which can effectively overcome the confounding biases and interference of reverse causalities in traditional observational researches with its unique advantages of the distributive randomness and timing priority of genetic variants. This article reviews the method of Mendelian randomization and its application in liver cancer， in order to provide new ideas for the research on causal association in liver cancer.

OBJECTIVE To explore the effects and mechanism of Bielong ruangan decoction on liver cancer model rats. METHODS Thirty-two male SD rats were randomly divided into control group， model group， and Bielong ruangan decoction low- dose and high-dose groups [6.84， 27.36 g/（kg·d）， by raw material]， with 8 rats in each group. Except for the control group， other groups were intraperitoneally injected with diethylnitrosamine （DEN） 50 mg/kg （once a week， for 16 consecutive weeks） to induce liver cancer model. At the 8th week of DEN injection， Bielong ruangan decoction low-dose and high-dose groups were orally administered with the corresponding medication， twice a day， until the 16th week. The general condition of rats in each group was observed during the experimental period. After the final administration， the body weight and liver mass were weighed， and the liver indexes were calculated； serum contents of alanine transaminase （ALT） and aspartate transaminase （AST） were determined； the appearance， pathological morphology and degree of fibrosis of liver were observed； Ishak scoring for liver fibrosis was performed； the mRNA and protein expressions of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 （NLRP3）， caspase-1， gasdermin D （GSDMD）， interleukin-18 （IL-18）， IL-1β in liver tissue were detected. RESULTS Compared with the model group， the general condition of rats （except for the low-dose group）， liver texture， surface nodules and tumors， inflammatory cell infiltration and abnormal cell amount were all improved in Bielong ruangan decoction low-dose and high-dose groups. Liver index （except for low-dose group）， Ishak score （except for low-dose group）， the serum contents of ALT and AST， as well as the mRNA and protein expressions of NLRP3， caspase-1， GSDMD， IL-18 and IL-1β in liver tissue were reduced significantly （P＜0.05）， with some of above indicators in high-dose group being significantly lower than those in low-dose group （P＜0.05）. CONCLUSIONS Bielong ruangan decoction can inhibit the progression of liver cancer in rats and reduce liver damage. Its mechanism of action may be related to the inhibition of the NLRP3/caspase-1/GSDMD signaling pathway and the improvement of inflammatory response.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.