BACKGROUND:The granulosa cell state greatly affects the quality of oocytes.As the increased proportion of aged women in assisted reproduction,granulosa cells from patients of different ages are examined at transcript levels in order to better assess oocyte quality. OBJECTIVE:To investigate the expression changes of mRNA and long non-coding RNA in granulosa cells from the young and aged patients. METHODS:Waste cumulus granulosa cells obtained during the assisted reproductive cycle.Young group was in 25-35 years old,and the aged group was in 38-45 years old.Granulosa cells collected from the young and aged females were analyzed using RNA sequencing,three replications for each group. RESULTS AND CONCLUSION:(1)The RNA sequencing analysis results showed that the average sequencing volume of samples was more than 14 G;the data quality Q30 after quality control was more than 93%;the average mapping rate of data was 98.4%,which indicates that the data had good quality.(2)The results of principal component analysis and correlation analysis showed that the samples of the aged group and the young group could be clearly distinguished.(3)The difference analysis results showed that compared with the young group,a total of 410 differentially expressed mRNAs were detected in the aged group(167 up-regulated and 243 down-regulated).GO analysis results showed that the down-regulated genes were mainly enriched in regulatedexocytosis and the emetabolicprocess.GSEA analysis results showed that secretion-related pathways in the aged group were down-regulated.(4)Compared with the young group,662 differentially expressed long non-coding RNAs were detected in the aged group,and 1 772 protein-coding genes were directly regulated by these long non-coding RNAs,among which 59 genes overlaped with differentially expressed genes.The results showed that the expression of secretion-related genes and pathways in granulosa cells of the aged group was down-regulated,thus affecting oocyte quality.At the same time,these down-regulated genes were regulated by long non-coding RNA.Therefore,the expression of long non-coding RNA might be related to age.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

Castleman disease(CD)is a rare lymphoproliferative disorder with an unclear etiology and pathogenesis.This article reports a case of unicentric Castleman disease(UCD)occurring in the submandibular region.The patient presented with a painless submandibular mass.Imaging studies revealed a well-defined,hypervascular lesion,and histopathological examination confirmed the diagnosis of plasma cell-type CD.Submandibular CD is exceptionally rare and can be easily misdiagnosed as lymphadenitis or lymphoma,necessitating a com-bination of imaging and pathological examinations for differential diagnosis.This case report provides valuable insights into diagnosis and management of CD.

Castleman disease(CD)is a rare lymphoproliferative disorder with an unclear etiology and pathogenesis.This article reports a case of unicentric Castleman disease(UCD)occurring in the submandibular region.The patient presented with a painless submandibular mass.Imaging studies revealed a well-defined,hypervascular lesion,and histopathological examination confirmed the diagnosis of plasma cell-type CD.Submandibular CD is exceptionally rare and can be easily misdiagnosed as lymphadenitis or lymphoma,necessitating a com-bination of imaging and pathological examinations for differential diagnosis.This case report provides valuable insights into diagnosis and management of CD.

OBJECTIVE To explore the colonization rates of carbapenem-resistant gram-negative bacteria(CRGNB)in intestinal tracts of intensive care unit(ICU)patients and analyze the influencing factors.METHODS The ICU patients were recruited from a three-A general hospital of Shanghai from Jan.2024 to Dec.2024,an ac-tive screening was carried out for intestinal tract CRGNB,and the detection rates of 5 types of carbapenems resist-ance genes(CRGs)in the CRGNB strains were observed.The baseline data and hospitalization data before the ac-tive screening were collected from the patients,logistic regression analysis was performed for the influencing fac-tors for the colonization of CRGNB.The effects of targeted infection control measures on length of ICU stay,hos-pitalization cost and prognosis were observed and compared before and after the active screening was carried out.RESULTS A total of 748 patients were included in the active screening,and the colonization rate of CRGNB in intestinal tracts was 11.50％(86/748).Among the CRGs,the detection rate of blaNDM was the highest(6.82％),followed by blaIMP(4.55％)and blaKPC(2.54％).The result of logistic regression analysis showed that,with an increase of 1 each day of hospital stay before the admission to ICUs,the risk was increased by 1.055 times(OR=1.055,95％CI:1.030 to 1.081,P＜0.001),the risk was 0.442 times the use of as aminoglycosides as the use of β-lactams(OR=0.442,95％CI:0.244 to 0.801,P=0.007).The targeted infection control measures that were taken after the active screening shortened the length of hospital stay(Z=-3.514,P＜0.001),reduce the hospitalization cost(Z=3.030,P=0.002)and improve the prognosis of the patients(x2=7.470,P=0.006).CONCLUSIONS The colonization rates of CRGNB in intestinal tracts are high among the ICU patients.It is necessary to reduce the length of hospital stay prior to ICU and strengthen the surveillance of drug-resistant strains and management of antibiotics.

This document targets digital human metabolic chamber platforms and specifies construction standards and testing protocols covering the full lifecycle of " build-test-operate." It encompasses chamber engineering and environmental control, digital platform and cybersecurity architecture, metabolic measurement and multimodal data acquisition, as well as quantitative system performance and data quality indicators with verifiable acceptance tests. By standardizing architecture, interfaces, and quality control, the specification enables multicenter data interoperability and harmonized quality management, providing high-quality, verifiable, and traceable infrastructure to support precision metabolism research and clinical translation in China.

Kounis syndrome is an acute coronary syndrome triggered by an allergic reaction, which is clinically rare and frequently subject to misdiagnosis or missed diagnosis. This article presents a case report of a 70-year-old male patient who developed a rash, pruritus, and chest pain following colon polyp resection. Coronary angiography revealed occlusion of the left anterior descending artery, and blood flow was restored after stent implantation. However, the patient experienced recurrent symptoms accompanied by loss of consciousness. Drug skin tests confirmed positive reactions to chlorhexidine and fondaparinux sodium, leading to a diagnosis of type Ⅱ Kounis syndrome. By avoiding allergenic drugs and combining antihistamines with symptomatic treatment to correct myocardial ischemia, the patient′s clinical symptoms significantly improved, and he eventually recovered and was discharged from the hospital. This case underscores the importance of maintaining vigilance for this syndrome in patients with allergies accompanied by chest pain and promptly identifying and avoiding allergens.

Objective To explore the effects of chronic stress and stress cessation on hypothalamic appetite regulators in mice,and to explore the stress-dependent mechanism of appetite change.Methods A total of 32 male C57BL/6 mice were randomly divided into control(Ctrl)group(n=16)and chronic unpredictable mild stress(CUMS)group(n=16).The mice in the CUMS group were given CUMS to establish the stress model,and those in the Ctrl group were fed normally.The food intake and weight of mice were recorded.The CUMS model was verified through tail suspension experiments and forced swimming experiments.Eight mice in the Ctrl group and 8 mice in the CUMS group were randomly sacrificed at the 12th week.The Ctrl group was re-grouped into the cessation-control(C-Ctrl)group(n=8),the CUMS group was re-grouped into the cessation-stress(C-CUMS)group(n=8),and the mice were sacrificed at the 15th week.The mRNA and protein levels of appetite-regulating factors,including orexin 1 receptor(OX1R),leptin receptor(LEPR)and agouti-related protein(AgRP)in the hypothalamus,were detected by quantitative polymerase chain reaction and immunochemistry.Results From week 2 to week 11 of stress,the food intake of the mice in the CUMS group was significantly higher than that in the Ctrl group(all P＜0.05),while there was no significant difference in body weight between the 2 groups within 11 weeks(all P＞0.05).Compared with the Ctrl group,the immobility durations of forced swimming and tail suspension of the CUMS group were markedly longer after 11 weeks(both P＜0.01),indicating successful modeling.AgRP and OX1R mRNA expression in the hypothalamus of the CUMS group was significantly increased(both P＜0.01),while LEPR mRNA expression was significantly decreased(P＜0.01);AgRP protein in the hypothalamic arcuate nucleus of the CUMS group was significantly higher than that of the Ctrl group(P＜0.05),and LEPR protein was markedly lower than that of the Ctrl group(P＜0.01).However,after 3 weeks of stress cessation,the C-CUMS group had less food intake and lower body weight than the C-Ctrl group(both P＜0.05).The LEPR mRNA of the C-CUMS group was significantly increased(P＜0.01),while AgRP and OX1R mRNA were not significantly different(both P＞0.05).There was no significant difference in AgRP protein levels between the C-CUMS group and the C-Ctrl group(P＞0.05),while LEPR protein level of the C-CUMS group was significantly higher than that of the C-Ctrl group(P＜0.01).Conclusion CUMS can lead to increased appetite in mice,which may involve the functional regulation of LEPR and AgRP.After the stress cessation,the appetite decreases,which may involve the functional regulation of LEPR.

Probiotics are natural systems bridging synthetic biology, physical biotechnology, and immunology, initiating innate and adaptive anti-tumor immune activity. We previously constructed an all-in-one engineered food-grade probiotic Lactococcus lactis (FOLactis) which could boost the crosstalk among different immune cells such as dendritic cells (DCs), natural killer cells, and T cells. Herein, considering the limited clinical efficacy of naked personalized neoantigen peptide vaccines, we decorate FOLactis with tumor antigens by employing a Plug-and-Display system comprising membrane-inserted peptides. Intranodal injection of FOLactis coated with neoantigen peptides (Ag-FOLactis) induces robust DCs presentation and neoantigen-specific cellular immunity. Notably, Ag-FOLactis not only triggers a 45-fold rise in the quantity of locally reactive neoantigen-specific T cells but also induces epitope spreading in both subcutaneous and metastatic tumor-bearing models, leading to potent inhibition of tumor growth. These findings imply that Ag-FOLactis represents a powerful platform to rapidly and easily display antigens, facilitating the development of a bio-activated platform for personalized therapy.