ObjectiveTo study on the different therapeutic effects and potential mechanisms of Rhei Radix et Rhizoma（RH） before and after steaming with wine on constipation model mice. MethodsFifty-four male ICR mice were randomly divided into control group， model group， lactulose group（1.5 mg·kg^-1）， high， medium and low dose groups of RH and RH steaming with wine（PRH）（8， 4， 1 g·kg^-1）. Except for the control group， the constipation model was replicated by gavage of loperamide hydrochloride（6 mg·kg^-1） in the other groups. After 2 weeks of modeling， each administration group was gavaged with the corresponding dose of drug solution， and the control and model groups were given an equal volume of normal saline， 1 time/d for 2 consecutive weeks. After administration， the feces were collected for 16S rRNA sequencing， the levels of gastrin（GAS）， motilin（MTL）， interleukin-6（IL-6）， γ-interferon（IFN-γ） in the colonic tissue were detected by enzyme-linked immunosorbent assay（ELISA）， the histopathological changes of colon were observed by hematoxylin-eosin（HE） staining， flow cytometry was used to detect the proportion changes of CD4⁺， CD8⁺ and regulatory T cell（Treg） in peripheral blood. ResultsCompared with the control group， the model group showed significantly decrease in fecal number in 24 h， fecal quality and fecal water rate（P<0.01）， the colon was seen to have necrotic shedding of mucosal epithelium， localized intestinal glands in the lamina propria were degenerated， necrotic and atrophied， a few lymphocytes were seen to infiltrate in the necrotic area in a scattered manner， the contents of GAS and MTL， the proportions of CD4⁺， CD8⁺ and Treg were significantly reduced（P<0.01）， the contents of IL-6 and IFN-γ were significantly elevated（P<0.05， P<0.01）. Compared with the model group， the fecal number in 24 h， fecal quality and fecal water rate of high-dose groups of RH and PRH were significantly increased（P<0.05， P<0.01）， the pathological damage of the colon was alleviated to varying degrees， the contents of GAS， MTL， IL-6 and IFN-γ were significantly regressed（P<0.05， P<0.01）， and the proportions of CD4⁺ and CD8⁺ were significantly increased（P<0.01）， although the proportion of Treg showed an upward trend， there was no significant difference. In addition， the results of intestinal flora showed that the number of amplicon sequence variant（ASV） and Alpha diversity were decreased in the model group compared with the control group， and there was a significant difference in Beta diversity， with a decrease in the relative abundance of Lactobacillus and an increase in the relative abundances of Bacillus and Helicobacter. Compared with the model group， the ASV number and Alpha diversity were increased in the high-dose groups of RH and PRH， and there was a trend of regression of Beta diversity to the control group， the relative abundance of Lactobacillus increased， and the relative abundances of Bacillus and Helicobacter decreased. ConclusionRH and PRH can improve dysbacteriosis， promote immune system activation， inhibit the release of inflammatory factors for enhancing the gastrointestinal function， which may be one of the potential mechanisms of their therapeutic effect on constipation.

Objective: In in vitro systems for differentiating and expanding natural killer (NK) cells, feeder cells provide essential cell-cell contact and paracrine signals that drive precursor proliferation and terminal maturation. However, existing xenogeneic feeder cells or tumor-derived genetically modified feeder cells pose risks of residual immunogenicity and malignant transformation, limiting clinical use. This study aims to develop a humanized mesenchymal-like stromal cell (hematopoietic organoid-derived human mesenchymal stromal cells, HO-hMSCs) derived from iPSC-based hematopoietic organoids, and elucidate its mechanisms of NK-supportive activity to enable a safe, efficient platform for clinical-grade NK cell production. Methods: Human induced pluripotent stem cells (iPSCs) were differentiated into hematopoietic organoids, from which HO-hMSCs were isolated. Flow-cytometric phenotyping and bulk RNA-sequencing were performed to compare HO-hMSCs with umbilical cord-derived MSCs (UC-hMSCs). The effect of HO-hMSCs on NK cell differentiation efficiency (CD3 CD56 ) and effector maturation (CD16 expression) were assessed by co-culture experiments, using UC-hMSCs as control. Results: 1) Hematopoietic organoid induction and NK differentiation: iPSCs were induced to form hematopoietic organoids using cytokine cocktails, which further differentiated into high-purity CD45 CD56 NK cells [(82.8%±12.07)% efficiency on day 21]. 2) HO-hMSC characteristics: HO-hMSCs exhibited upregulated expression of Notch pathway ligands (DLL4, JAG1, 4.06-8.04-fold), homeobox genes (HOXA3, HOXA5, log FC=1.28 and 1.44), and key regulators of NK development (GATA3, BCL11A) and cytokine receptors (IL7R, IL27RA, 6.76 to 13.34-fold increase). 3) Functional validation: Compared to UC-hMSCs, HO-hMSCs co-culture significantly enhanced NK cell proportion by 30.5% (P<0.05) and increased CD16 positivity (+20.5%). Conclusion: This study for the first time reveals that human hematopoietic organoid-derived HO-hMSCs possess potent hematopoietic niche-supportive activity. It provides a humanized, feeder-free platform for robust clinical-grade NK cell production and expands the translational utility of organoid technologies in cell therapy.

Flexible conductive fibers have been widely applied in wearable flexible sensing. However, exposed wearable flexible sensors based on liquid metal (LM) are prone to abrasion and significant conductivity degradation. This study presented a high-sensitivity LM conductive fiber with integration of strain sensing, electrical heating, and thermochromic capabilities, which was fabricated by coating eutectic gallium-indium (EGaIn) onto spandex fibers modified with waterborne polyurethane (WPU), followed by thermal curing to form a protective polyurethane sheath. This fiber, designated as Spandex/WPU/EGaIn/Polyurethane (SWEP), exhibits a four-layer coaxial structure: spandex core, WPU modification layer, LM conductive layer, and polyurethane protective sheath. The SWEP fiber had a diameter of (458.3 ± 10.4) μm, linear density of (2.37 ± 0.15) g/m, and uniform EGaIn coating. The fiber had excellent conductivity with an average value of (3 716.9 ± 594.2) S/m. The strain sensing performance was particularly noteworthy. A 5 cm × 5 cm woven fabric was fabricated using polyester warp yarns and SWEP weft yarns. The fabric exhibited satisfactory moisture permeability [(536.06 ± 33.15) g/(m ²·h)] and maintained stable thermochromic performance after repeated heating cycles. This advanced conductive fiber development is expected to significantly promote LM applications in wearable electronics and smart textile systems.
Wearable Electronic Devices ; Polyurethanes/chemistry* ; Electric Conductivity ; Gallium/chemistry* ; Metals/chemistry*

OBJECTIVES: To explore the bioactive components in Jiawei Xiaoyao Pills (JWXYP) and their mechanisms for alleviating depression-like behaviors. METHODS: The active compounds, key targets, and pathways of JWXYP were identified using TCMSP and TCMIP databases. Thirty-six SD rats were randomized equally into 6 groups including a control group and 5 chronic unpredictable mild stress (CUMS)-induced depression groups. After modeling, the 5 model groups were treated with daily gavage of normal saline, 1.8 mg/kg fluoxetine hydrochloride (positive control drug), or JWXYP at 1.44, 2.88, and 4.32 g/kg. The depression-like behaviors of the rats were evaluated using behavioral tests, and pathological changes in the liver and hippocampus were examined with HE staining. The biochemical indicators in the serum and brain tissues were detected using ELISA. Serum metabolomics analysis was performed to identify the differential metabolites using OPLS-DA, and gut microbiota changes were analyzed using 16S rDNA sequencing. RESULTS: Network pharmacology revealed that menthone and paeonol in JWXYP were capable of penetrating the blood-brain barrier to regulate inflammatory pathways and protect the nervous system. In the rat models subjected to CUMS, treatment with JWXYP significantly improved body weight loss, sucrose preference and open field activities, reduced liver inflammation, alleviated structural changes in the hippocampal neurons, decreased serum levels of TNF‑α, IL-1β, IL-6 and LBP, and increased 5-HT and VIP concentrations in the serum and brain tissue, and these effects were the most pronounced in the high-dose group. Metabolomics analysis showed changes in such metabolites as indole-3-acetamide and acetyl-L-carnitine in JWXYP-treated rats, involving the pathways for bile acid biosynthesis and amino acid metabolism. 16S rDNA analysis demonstrated increased gut microbiota diversity and increased abundance of Lactobacillus species in JWXYP-treated rats. CONCLUSIONS JWXYP alleviates depression-like symptoms in rats by regulating the neurotransmitters, inhibiting inflammation and oxidation, and modulating gut microbiota.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Gastrointestinal Microbiome/drug effects* ; Rats, Sprague-Dawley ; Depression/drug therapy* ; Neurotransmitter Agents/metabolism* ; Rats ; Inflammation ; Male ; Hippocampus ; Behavior, Animal/drug effects*

Objective:To investigate the potential effect and mechanism of docosahexaenoic acid(DHA) on hippocampal neuronal apoptosis and c-Jun N-terminal kinase(JNK) protein in rats with subarachnoid hemorrhage(SAH).Methods:Forty eight SPF-grade male SD rats were randomly divided into control group, sham group, model group and DHA intervention group according to the random number table method, with 12 rats in each group.The rats in model group and DHA group were injected with autologous blood(0.3 mL) into the optic chiasma to establish the SAH model.Rats in model group were intraperitoneally injected with DHA(35 mg/kg) 3 hours after SAH model establishment, rats in sham operation group were injected with 0.3 mL 0.9% sodium chloride solution into the optic chiasma, and rats in control group were fed normally.Neurobehavioral function of all rats was evaluated after 24 hours.The apoptosis of neuron was observed by TUNEL staining, and the expression of phosphorylated JNK(p-JNK)and apoptosis-related proteins Bax and Bcl-2 was observed by Western blot.Statistical analysis was performed using GraphPad Prism 7.0 software.One-way ANOVA was used for comparison among multiple groups, and Tukey test was used for further pairwise comparison.Results:(1)The differences in neurobehavioral function scores among the 4 groups of rats were statistically significant( F=103.60, P<0.05), the neurobehavioral function scores in model group(8.67±1.37) and DHA intervention group(13.67±1.51) were lower than that in control group(18.00±0.00) and sham group(17.67±0.52)( all P<0.05), while the neurobehavioral function score in DHA intervention group was higher than that in the model group( P<0.05).(2)The results of TUNEL staining showed that there were statistical differences in the number of hippocampal neuron apoptosis among the 4 groups( F=30.76, P<0.05), the number of hippocampal neuron apoptosis in model group(55.67±5.28) was higher than those in control group(25.83±7.06) and sham group(25.50±6.72) (both P<0.05), the number of hippocampal neuron apoptosis in DHA intervention group(35.17±5.78) was lower than that in model group.(3)The results of Western blot showed that there were no statistical differences in the Bax protein levels among the four groups( F=2.00, P>0.05).There were statistical differences in the expression levels of Bcl-2, p-JNK and Bax/Bcl-2 ratio among the 4 groups( F=8.48, 5.69, 5.39, all P<0.05).There was no statistical difference in Bcl-2, p-JNK protein levels and Bax/Bcl-2 ratio between the control group and sham group(all P>0.05).The p-JNK protein levels and Bax/Bcl-2 ratio in model group ((1.93±0.25), (2.05±0.86)) were higher than those in sham group ((1.42±0.33), (1.05±0.26)) (both P<0.05), the Bcl-2 protein level in model group (1.04±0.23) was lower than that in sham group (1.61±0.16) ( P<0.05).The p-JNK protein level and Bax/Bcl-2 ratio in DHA intervention group((1.43±0.33), (1.19±0.30)) were lower than those in model group(both P<0.05), the Bcl-2 protein level in DHA intervention group(1.42±0.28) was higher than that in model group( P<0.05). Conclusion:DHA can reduce neuronal apoptosis, inhibit the activation of p-JNK and improve neurological function of SAH model rats.

Dysfunction of nuclear receptors(NRs),which are critical for maintaining renal lipid homeostasis,largely contributes to the progression of diabetic nephropathy(DN)by promoting renal lipid accumulation.However,there are few thera-pies in modern medicine that effectively inhibit re-nal lipid accumulation by modulating NRs.In this review,we first summarize recent advances in NRs-mediated renal lipid accumulation in DN.Then,we summarize recent studies on the effects and mech-anisms of Chinese herbal medicines on DN and highlight the effects of Chinese herbal medicines on renal lipid accumulation via NRs.In addition,the advantages and limitations of traditional Chi-nese medicine in the treatment of DN via NRs-me-diated renal lipid accumulation are analyzed,and the modulation of renal lipid accumulation via NRs is emphasized as a promising therapeutic strategy for the treatment of DN.

Objective:To explore the mechanism of EPO on activating receptors of NK cells in rats with diabetic nephropathy based on HMGB1/Beclin-1 signaling pathway.Methods:Forty SPF male SD rats were randomly divided into normal group(A),model group(B),metformin group(C)and EPO group(D),with 10 rats in each group.Diabetic nephropathy modeling was performed on groups B,C and D using high-fat diet and streptozotocin.After successful modeling,group C was given metformin 100 mg/kg by intra-gastric administration,and group D was intraperitoneal injection of EPO 100 U/kg.Group A and B were given normal saline at the same volume by intragastric administration simultaneously.After successful modeling,the remaining 20 rats were randomly divided into group E and group F.Group E was given 30 mg/kg HMGB1 inhibitor by intragastric administration,group F was given 30 mg/kg HMGB1 inhibitor by intragastric administration and 100 U/kg EPO by intraperitoneal injection.HK-2 cells were taken and divided into high glucose+HK-2 cells(HH)group,EPO+high glucose+HK-2 cells(EH)group,glycyrrhizin L+high glucose+HK-2 cells(LH)group,and cultured with glucose for cell experiment.Blood glucose was detected by glucose analyzer,biochemical indexes were de-tected by automatic biochemical analyzer,and renal pathological morphology of rats was detected by PAS staining.Expression of HMGB1/Beclin-1 protein was detected by Western blot,and expressions of NK cell activated receptors were detected by RT-PCR and immunohistochemistry.Results:Compared with group A,blood glucose,24 h urine volume,blood glucose curve,BUN,Scr,UAlb contents in blood and urine,mRNA and protein expressions of NKp30,NKp44,NKp46 in group B were significantly increased(P<0.05),while body weight was significantly decreased(P<0.05).Compared with group B,blood glucose,24 h urine volume,blood glucose curve,BUN,Scr,UAlb contents in blood and urine,mRNA and protein expression of NKp30,NKp44 and NKp46 in group C and group D were significantly decreased(P<0.05),while body weight was significantly increased(P<0.05),renal pathology was also significantly improved,the changes of group D was significantly higher than that of group C(P<0.05).Compared with group A,expression of HMGB1/Beclin-1 protein in renal tissues of rats in group B was significantly increased(P<0.05),and expression of HMGB1/Beclin-1 protein in renal tissues of rats in groups C,D,E and F was significantly decreased(P<0.05),expression of HMGB1/Beclin-1 protein in group D was significantly decreased compared with group C(P<0.05),there was no significant difference between group E and group D,and expression of HMGB1/Beclin-1 protein in group F was significantly decreased compared with group E.HMGB1/Beclin-1 protein expression was positively correlated with NKp30,NKp44 and NKp46 mRNA(P<0.05).Compared with HH group,proliferation rates of EH and LH groups were significantly increased(P<0.05),while apoptosis rate and HMGB1/Beclin-1 protein expression were significantly decreased(P<0.05),there was no significant difference between LH group and EH group(P>0.05).Conclusion:EPO can effectively reduce expressions of NK cell activated receptors and inhibit HMGB1/Beclin-1 signaling path-way in diabetic nephropathy rats,suggesting that EPO may exert its effect by regulating NK cell activated receptors and HMGB1/Beclin-1 signaling pathway.NK cell activation receptors expressions are positively correlated with HMGB1/Beclin-1 signaling pathway.

Objective:To analyze three reconstruction techniques and mid-term clinical outcomes of hip revision for acetabular bone defect after total hip arthroplasty (THA).Methods:This is a retrospective case series study. Included in the study were 109 patients (109 hips) with acetabular bone defect after THA reconstructions in hip revisions from January 2015 to December 2021 in the Senior Department of Orthopaedics, the Forth Medical Center of Chinese People′s Liberation Army General Hospital and the Department of Orthopaedics, the First Medical Center of Chinese People′s Liberation Army General Hospital. According to the preoperative simulated surgeries and different bone defect reconstruction techniques, patients were divided into a normal cup group, an augment group or a triflange group,respectively. There were 54 patients (54 hips) in the normal cup group, reconstructed with the uncemented porous metal cup (including Jumbo cup), with 23 males and 31 females, aged (59.6±9.9) years (range:32 to 76 years); 44 patients (44 hips) in the augment group, reconstructed with the individualized three-dimensional (3D) printed porous metal augment and uncemented porous metal cup, with 18 males and 26 females, aged (52.8±13.6) years(range:17 to 76 years); 11 patients (11 hips) in the triflange group, reconstructed by the individualized 3D printed porous metal triflange cup, with 5 males and 6 females, aged (59.4±11.2) years (range: 43 to 78 years). Radiographic results, including rotation center height, rotation center offset, and leg length discrepancy (LLD) and clinical results, including Harris hip score (HHS) and visual analogue scale(VAS) were evaluated outpatient at 3, 6, 12 months after the operation and annually thereafter. The last follow-up was completed in March 2024, and all parameters at the last follow-up and before the operation were compared. Paired sample t test and repeated measurement ANOVA were used for the radiographic and clinical parameters before and after the operation. Results:All hip revisions for patients with acetabular bone defect after THA were completed and followed for more than two years. The follow-up time of the normal cup group was (6.5±1.7) years (range: 2.8 to 9.3 years), and that of the augment group was (6.0±1.3) years (range: 3.5 to 9.0 years). The follow-up time of the triflange group was (2.8±0.6) years (range: 2.0 to 3.8 years). At the last follow-up, the rotation center height, rotation center offset and LLD of 54 hips in the normal cup group were (24.2±5.6) mm, (29.1±5.5) mm and (4.6±3.3) mm, respectively, and the rotation center height and LLD were significantly lower than those of the preoperative hips ( t=9.671, P<0.01; t=6.073, P<0.01). In the augment group, the rotational center height, the rotation center offset and the LLD of 44 hips were (22.4±9.0) mm, (25.4±5.5) mm and (6.0±4.0) mm, respectively, which were significantly lower than those of the preoperative hips ( t=9.071, P<0.01; t=11.345, P<0.01; t=4.927, P<0.01). In the triflange group, the rotational center height, the rotation center offset and LLD of 11 hips were (22.7±6.0) mm,(30.9±8.0) mm and (5.3±2.2) mm, respectively, and the rotation center height and LLD were significantly lower than those of the preoperative hips ( t=2.716, P=0.022; t=6.226, P<0.01). At the last follow-up, fractures occurred in 3 patients and dislocation occurred in 1 patient in the normal cup group, and fracture reduction and closed reduction were administered under anesthesia, respectively. In the augment group, dislocation occurred in 1 patient and open reduction under anesthesia was performed. The HHS and VAS of the three groups improved significantly after surgery and the differences were statistically significant (all P<0.01). There was no complication in the triflange group. The X-ray at the last follow-up showed that all prostheses and augments were in stable positions and no loosening or migration was observed. Conclusions:For patients with acetabular bone defect after THA undergoing hip revisions, preoperative surgical simulation and rehearsal could help surgeons choose convenient and efficient reconstruction techniques. The targeted selection of Jumbo cup, individualized 3D printed metal augment, and customized triflange cup could achieve satisfactory clinical outcomes.

Objective Utilizing a specially engineered miR-144-GFP-H1 human embryonic stem cell(hESC)reporter line,this study leverages GFP fluorescence as an indicator of miR-144 expression to gauge the progression of erythropoiesis.The investigation is aimed at elucidating the potential roles of lncRNAs within the erythropoietic framework and conducting an initial assessment of their functional impact.Methods The miR-144/451-GFP-H1 cell line(hereafter referred to as 144-H1)was utilized for in vitro erythrocyte induction culture.The subpopulations of cells entering the erythropoiesis stage were characterized by the surface molecules CD71 and GPA.The GFP reporter gene of miR-144 served as a critical determi-nant to distinguish between GFP-positive cells(with a high propensity for erythropoiesis)and GFP-negative cells(with a low propensity for erythropoiesis).Transcriptome sequencing was performed on both groups to identify differentially ex-pressed long non-coding RNAs(lncRNAs).LncRNA entries with potential for validation were selected for preliminary func-tional verification.The CRISPR/Cas9 gene editing technique was employed to design functional interference strategies for the targeted lncRNAs,obtaining 144-H1 cell lines with knocked-out function of the specific lncRNAs.These knockout cell lines,along with non-knockout 144-H1 cell lines,were used for parallel erythrocyte induction culture to identify differential nodes.This approach preliminarily verified their impact on erythropoiesis in an in vitro development model.Results 1)The constructed 144-H1 cell line was capable of expressing GFP fluorescence upon entering the stage of in vitro erythrocyte in-duction,indicating the activation of miR-144/451.2)Within the CD71,GPA double-positive group,significant differences in lncRNA expression were observed between the GFP-positive and GFP-negative subpopulations.3)Gene editing strategies involving the deletion of sequence segments capable of effectively interfering with the function of multiple lncRNA entries were designed and verified for successful editing.In the knockout cell lines,parts of the lncRNA sequences were directly de-leted.4)In parallel validation experiments of erythrocyte induction culture,cell lines with LINC01569 knocked out exhibited significant differences in flow cytometric subpopulations and cell proliferation capabilities compared to the non-knockout cell lines:①The knockout cell lines showed sustained high expression of GFP fluorescence.②The proportion of the CD71-GPA double-positive group in the knockout cell lines continuously decreased during erythrocyte maturation.③No significant ex-pression of hemoglobin was observed in the knockout cell lines,lacking the characteristic red color.④The cell proliferation capability of the knockout cell lines was significantly lower than that of the non-knockout cell lines(P＜0.05).Conclusion The successful employment of the 144-H1 cell line facilitated an exploration into the potential functions of lncRNAs in e-rythropoiesis.This enables the design of more refined in vitro developmental experiments to enhance the precision in captu-ring lncRNA functions.Among the differentially expressed lncRNA entries,LINC01569 was preliminarily validated to play a regulatory role in erythropoiesis.The functional absence of LINC01569 severely impacts the normal differentiation and prolif-eration of erythrocytes.The specific regulatory mechanism of LINC01569 in erythropoiesis warrants further investigation and research.

Objective To investigate the current situation and health effects of indoor microbial pollution in Xi'an, to analyze the purification effect of air purifiers on indoor microbial pollution, and to provide reference for improving the indoor environment. Methods Through stratified random sampling, 20 families from rural areas and 20 families from upwind and downwind urban areas respectively were selected from Xi'an. Data was collected by questionnaire surveys and on-site environmental sampling. Non-parametric analysis and correlation analysis were used for statistical analysis. Results Overall, the standard-exceeding rate of total count of bacteria was 5.00%. The medians of the total count of bacteria and fungi were 312.50 cfu/m3 and 260.00 cfu/m³, respectively. In terms of health effects, the correlation between rhinitis and cold with total bacterial count was statistically significant (P<0.05), with the correlation coefficients of 0.182 and 0.223, respectively. Purification effect of air purifiers on microbial pollution was statistically significant (P<0.05). After opening for 2 hours, the total numbers of bacteria and fungi decreased significantly. Conclusion The occurrence risk of colds and rhinitis is increased by indoor microbial pollution. Air purifiers have a certain effect on decreasing the total number of bacteria and fungi. It is recommended to use air purifiers with high CADR of particulate matter, double-layer filter and sterilization and dehumidification function, and replace the filters regularly to reduce indoor microbiological contamination.