Bacillus coagulans can utilize the hydrolyzed carbon source of agricultural waste to produce lactic acid via a homofermentative pathway. However, a significant carbon source metabolic repression effect was observed when the strain metabolized mixed sugars (glucose and xylose), reducing the productivity of lactic acid. In this study, we obtained the fermentation conditions for the simultaneous utilization of the mixed sugars by B. coagulans by changing the ratio of glucose to xylose in the medium. Through transcriptome sequencing, several key genes responsible for xylose utilization were identified. The critical role of xylose isomerase (XylA, EC 5.3.1.5) in the synchronous utilization of glucose/xylose in B. coagulans was investigated via qRT-PCR (quantitative real-time polymerase chain reaction). Subsequently, the heterologous expression and characterization of the XylA-encoding gene (XylA) were conducted. It was determined that the gene encoded a protein composed of 440 amino acid residues. The secondary structure of the encoded protein was predominantly composed of α-helixes and random coils, while the higher structure of the protein was identified as a homotetramer. Then, XylA was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant protein Bc-XlyA was obtained with a molecular weight of approximately 50 kDa. The optimal pH and temperature of Bc-XylA were 8.0 and 60 ℃, respectively, and Mn²⁺, Mg²⁺, and Co²⁺ had positive effects on the activity of Bc-XlyA. The present study provides scientific data on the molecular modification of B. coagulans, offering theoretical support for the efficient utilization of xylose in the strain.
Xylose/metabolism* ; Cloning, Molecular ; Bacillus coagulans/enzymology* ; Aldose-Ketose Isomerases/metabolism* ; Fermentation ; Bacterial Proteins/metabolism* ; Glucose/metabolism*

Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC)assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.

Objective·To determine hyaluronan(HA)expression in the endocrine-resistant microenvironment of estrogen receptor-positive(ER+)breast cancer and elucidate its impact on the acquired resistance.Methods·Chemiluminescent immunoassay was used to quantify HA levels in the culture supernatants of fulvestrant-resistant breast cancer cells.An immunofluorescence(IF)assay was performed to visualize the colocalization of CD44 and HA in MCF7/FulR cells.Using an established adaptive endocrine-resistant breast cancer mouse model,HA expression in resistant breast cancer tissues was assessed by immunohistochemistry(IHC)assay.Single-cell RNA sequencing(scRNA-seq)and RNA sequencing(RNA-seq)were conducted to examine transcriptomic profiles and alterations in HA-related genes in resistant breast cancer cells.Flow cytometry(FCM)was utilized to measure the proportion of CD44+CD24-cells in MCF7/FulR.The correlation between HA synthesis genes and cell stemness was investigated in clinical ER+breast cancers from GEO data sets.Hyaluronidase(HAase)treatment was applied to remove the"HA coat",and RT-qPCR and Western blotting analysis were carried out to monitor changes in stemness-related molecules.CCK-8 assays,flow cytometry(FCM),and Hoechst 33258 staining were performed to determine changes in apoptosis and fulvestrant efficiency after HAase treatment.Results·IF results revealed that compared with MCF7 cells,the"HA coat"on the surface of MCF7/FulR cells was significantly thickened.IHC demonstrated markedly increased HA retention in fulvestrant-resistant mouse breast cancer tissues.ScRNA-seq and RNA-seq analyses indicated elevated expression of stemness-related genes and HA synthesis-associated genes in fulvestrant-resistant breast cancer cells.Correlation analysis revealed a positive association between HA synthesis and cancer stemness in ER+breast cancer.IF and RT-qPCR results demonstrated that removing the HA coating from the surface of MCF7/FulR cells led to a significant reduction in the expression of stemness-related molecules;concurrently,CCK-8 assays,FCM analysis,and Hoechst 33258 staining revealed that"HA coat"clearance reduced MCF7/FulR'tolerance to fulvestrant and increased apoptosis.Conclusion·Endocrine-resistant breast cancer cells develop an enriched"HA coat",which promotes stemness in fulvestrant-resistant tumors.Disruption of this HA coat through HAase treatment effectively reduces cell stemness,induces apoptosis,and re-sensitizes breast cancer cells to fulvestrant.

By sorting out the literature related to constitution and referring to the concept and method of "element"， many ancient and modern theories of constitution are deconstructed into multi-dimensional "constitution elements". These constitution elements are divided into two levels： basic and derivative. The basic level is the most basic element of constitution， including zang-fu organs/body and material. The zang-fu organs/body dimension focuses on the fixed functional units in the human body， and the material dimension focuses on the rheology functional units of human body. The derivative level is the secondary physiological or psychological state in life activities， including emotion and qi transformation. The emotional dimension focuses on the state of human psychological activities， and the qi transformation dimension focuses on the state of human physiological activities in cold， heat， dampness， and dryness. Based on this， we can further study the influence of factors such as age， gender， region， and social class， on the combination rule of "constitution element"， so as to form a more comprehensive classification program of constitution.

Objective:To explore the application value of shear wave elastography (SWE) in male anterior urethral stenosis.Methods:A total of 40 male patients with anterior urethral stenosis admitted to the Sixth People′s Hospital Affiliated to Medical College of Shanghai Jiaotong University from September 2022 to March 2023 were collected. SWE examination was performed before operation to measure the elasticity of urethra and surrounding normal tissues, and to evaluate the location and length of urethral stenosis. When the patient underwent urethral repair and reconstruction surgery, the lesion site of urethral stenosis was measured during the operation to confirm the actual location and length of the lesion during the operation, and the lesion site and length of the urethral stenosis were compared with the preoperative SWE assessment. The specimen of urethral lesion excised during the operation was sent for pathological examination to further confirm whether the tissue was pathological scar tissue.Results:The elastic value of urethral stricture in SWE [(32.9±6.2)kPa] was higher than that of normal urethra [(21.5±4.3)kPa], and the difference was statistically significant ( P<0.05). The length of urinary tract stricture measured by preoperative SWE examination [(3.51±1.09)cm] was basically the same as that of urethral stricture measured during operation [(3.51±1.10)cm], with no statistical significance ( P=0.825). The pathological examination proved that the surgically resected tissue was indeed pathological scar tissue, and the pathological results of the site of urethral stenosis determined by the preoperative SWE examination were 100% consistent with those of the postoperative stenosis. Conclusions:SWE can well evaluate the length of male anterior urethral stenosis and the location of the diseased urethra before surgery, and provide accurate guidance for the scope of urethral lesion and pathological scar resection during surgery.

Background/Aims: The shortage of donor liver hinders the development of liver transplantation. This study aimed to clarify the poor outcomes of functionally marginal liver grafts (FMLs) and provide evidence for the improvement of ischemia-free liver transplantation (IFLT) after FML transplantation. Methods: Propensity score matching was used to control for confounding factors. The outcomes of the control group and FML group were compared to demonstrate the negative impact of FMLs on liver transplantation patients. We compared the clinical improvements of the different surgical types. To elucidate the underlying mechanism, we conducted bioinformatic analysis based on transcriptome and single-cell profiles. Results: FMLs had a significantly greater hazard ratio (HR: 1.969, P=0.018) than did other marginal livers. A worse 90-day survival (Mortality: 12.3% vs. 5.0%, P=0.007) was observed in patients who underwent FML transplantation. Patients who received FMLs had a significant improvement in overall survival after IFLT (Mortality: 10.4% vs 31.3%, P=0.006). Pyroptosis and inflammation were inhibited in patients who underwent IFLT. The infiltration of natural killer cells was lower in liver grafts from these patients. Bulk transcriptome profiles revealed a positive relationship between IL-32 and Caspase 1 (R=0.73, P=0.01) and between IL-32 and Gasdermin D (R=0.84, P=0.0012). Conclusions FML is a more important negative prognostic parameter than other marginal liver parameters. IFLT might ameliorate liver injury in FMLs by inhibiting the infiltration of NK cells, consequently leading to the abortion of IL-32, which drives pyroptosis in monocytes and macrophages.

Alcoholic steatohepatitis (ASH) is a liver disease characterized by steatosis, inflammation, and necrosis of the liver tissue as a result of excessive alcohol consumption. Pregnane X receptor (PXR) is a xenobiotic nuclear receptor best known for its function in the transcriptional regulation of drug metabolism and disposition. Clinical reports suggested that the antibiotic rifampicin, a potent human PXR activator, is a contraindication in alcoholics, but the mechanism was unclear. In this study, we showed that the hepatic expression of fatty acid binding protein 4 (FABP4) was uniquely elevated in ASH patients and a mouse model of ASH. Pharmacological inhibiting FABP4 attenuated ASH in mice. Furthermore, treatment of mice with the mouse PXR agonist pregnenolon-16α-carbonitrile (PCN) induced the hepatic and circulating levels of FABP4 and exacerbated ASH in a PXR-dependent manner. Our mechanism study established FABP4 as a transcriptional target of PXR. Treatment with andrographolide, a natural compound and dual inhibitor of PXR and FABP4, alleviated mice from ASH. In summary, our results showed that the PXR-FABP4 gene regulatory axis plays an important role in the progression of ASH, which may have accounted for the contraindication of rifampicin in patients of alcoholic liver disease. Pharmacological inhibition of PXR and/or FABP4 may have its promise in the clinical management of ASH.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate the prevalence of frailty among elders aged ≥60 years in the Beijing community and analyze the risk factors of frailty.Methods:This is a cross-sectional study. A total of 1936 older persons in Baizhifang Community in Beijing were included between May and September 2023. Their frailty was evaluated by Fried Frailty Phenotype Scale. Mini Nutritional Assessment-Short Form (MNA-SF), Short Physical Performance Battery (SPPB), and Geriatric Depression Scale (GDS) were used to evaluate nutritional status, physical function, and depression, respectively. The prevalence and risk factors of frailty among the elderly in the community were analyzed.Results:A total of 168 individuals met the criteria for frailty, and the prevalence of frailty was 8.7%. Compared to the non-frail group, the frail group was predominantly female (71.4% vs 62.2%; χ 2=5.59, P=0.018), older [(72.1±6.6) vs (69.9±6.0) ages; t=-5.25, P<0.001], unmarried (1.8% vs 0.8%; χ 2=8.60, P=0.014), with lower education levels (10.7% vs 3.5%; χ 2=23.38, P<0.001), poorer self-assessed health (22.6% vs 6.5%; χ 2=69.80, P<0.001), lower MNA-SF scores [(12.2±2.2) vs (13.0±1.4) scores; t=6.29, P<0.001], lower SPPB scores [(8.4±1.9) vs (10.2±1.6) scores; t=14.62, P<0.001], higher GDS scores [(4.0±3.8) vs (1.9±2.1) scores; t=-11.48, P<0.001], and worse vision, hearing, and olfactory functions (48.2% vs 34.1%, 36.9% vs 23.0%, 9.5% vs 3.0%; χ 2=13.37, 16.11, 40.58, all P<0.001). They were also more likely to suffer from hypertension (70.8% vs 56.7%; χ 2=12.52, P<0.001), diabetes (42.3% vs 29.4%; χ 2=12.06, P<0.001), osteoarthritis (46.4% vs 30.3%; χ 2=18.39, P<0.001) and stroke (19.0% vs 13.3%; χ 2=4.28, P=0.039). Logistic regression analysis showed that frailty was independently associated with nutritional status ( OR=0.8, 95% CI: 0.7-0.9, P<0.001), depressive symptoms ( OR=1.1, 95% CI: 1.1-1.2, P<0.001), olfactory function ( OR=2.1, 95% CI: 1.4-3.0, P<0.001; OR=2.9, 95% CI: 1.4-5.9, P=0.003), and physical function ( OR=0.6, 95% CI: 0.6-0.7, P<0.001) after adjusting for covariates. Conclusions:The prevalence of frailty among elderly people in community is relatively high. Frailty in community elders is associated with physical function, psychological function, nutritional status, and olfactory function decline.

Objective:To analyze mobility limitation condition and its relationship with cognitive function in community-dwelling elderly.Methods:In this cross-sectional study, a total of 1 935 older adults aged≥60 years were recruited from May to September 2023 in Beijing Baizhifang Community Health Service Center and its five subordinate health service stations (Baizhifang Hutong health service station, Nancaiyuan community health service station, Youanmen community health service station, Younei West Street health service station and Shuanghuaili community health service station). The Short Physical Performance Battery (SPPB) was used to assess the mobility capacity of the elderly, and the elderly were divided into two groups with SPPB, the elderly with a SPPB≤9 points were grouped into mobility limitation group (645 cases), and the ones with a SPPB≥10 points were considered with normal mobility capacity (1 290 cases). The cognitive function of the older adults was assessed with the mini-mental state examination; and the gender, age, calf circumference, history of chronic disease, frailty status, cognitive function, nutritional status, depression status, hearing and vision condition of the two groups were compared with χ2 test or Mann-Whitney U non-parametric test. Multiple logistic regression analysis was used to analyze the mobility limitation and its relationship with cognitive function in the older adults. Results:Mobility limitation was found in 33.33% (645/1 935) of community-dwelling older adults. The proportions of advanced age, female, hypertension, diabetes, coronary heart disease, stroke, chronic obstructive pulmonary disease, frailty, depression, cognitive impairment, hearing decline, and vision decline in the mobility limitation group were all significantly higher than those in the normal mobility capacity group (31.32% vs 13.41%, 69.92% vs 59.61%, 65.27% vs 54.03%, 33.80% vs 28.60%, 27.91% vs 19.53%, 17.83% vs 11.47%, 10.54% vs 7.36%, 13.18% vs 2.02%, 18.45% vs 6.59%, 14.73% vs 7.75%, 30.54% vs 20.31%, 45.58% vs 30.39%) (all P<0.05). Advanced age ( OR=2.542, 95% CI: 1.977-3.269), female ( OR=1.736, 95% CI: 1.390-2.167), stroke ( OR=1.426, 95% CI: 1.065-1.911), depression ( OR=2.292, 95% CI: 1.656-3.174), cognitive impairment ( OR=1.601, 95% CI: 1.154-2.220), frailty ( OR=5.199, 95% CI: 3.219-8.397) and vision decline ( OR=1.405, 95% CI: 1.124-1.756) were all positively correlated with the mobility limitation in the community-dwelling older adults (all P<0.05). Conclusion:The older adults in the community have a higher risk of mobility limitation, mobility limitation is a positive correlation factor of cognitive impairment.