Objective To investigate the effects of hyaluronic acid and proteoglycan-linked protein 1 (HAPLN1) secreted by synovial fibroblasts (FLS) on the polarization of macrophages (Mϕ) in rheumatoid arthritis (RA). Methods Human monocytic leukemia cells (THP-1) were differentiated into Mϕ, which were subsequently exposed to recombinant HAPLN1 (rHAPLN1). RA-FLS were transfected separately with HAPLN1 overexpression plasmid (HAPLN1^OE) or small interfering RNA targeting HAPLN1 (si-HAPLN1), and then co-cultured with Mϕ to establish a co-culture model. The viability of Mϕ was assessed using the CCK-8 assay, and the proportions of pro-inflammatory M1-type and anti-inflammatory M2-type Mϕ were analyzed by flow cytometry. Additionally, the expression levels of inflammatory markers, including interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), and inducible nitric oxide synthase (iNOS), were quantified using quantitative real-time PCR and Western blot analysis. Results The viability of Mϕ was increased in the rHAPLN1 group compared to the control group. Furthermore, both the M1/Mϕ ratio and inflammatory factor levels were elevated in the rHAPLN1 and HAPLN1^OE groups. In contrast, the si-HAPLN1 group exhibited a decrease in the M1/Mϕ ratio and inflammatory factor expression. Notably, the introduction of rHAPLN1 in rescue experiments further promoted Mϕ polarization towards the M1 phenotype. Conclusion HAPLN1, secreted by RA fibroblast-like synoviocytes (RA-FLS), enhances Mϕ polarization towards the M1 phenotype.
Humans ; Arthritis, Rheumatoid/genetics* ; Macrophages/immunology* ; Fibroblasts/metabolism* ; Phenotype ; Extracellular Matrix Proteins/genetics* ; Proteoglycans/genetics* ; Synovial Membrane/cytology* ; Tumor Necrosis Factor-alpha/genetics* ; Interleukin-1beta/genetics* ; Nitric Oxide Synthase Type II/genetics* ; Cell Differentiation ; Coculture Techniques ; THP-1 Cells

The catalytic activity of 3-mercaptopyruvate (3MP) sulfurtransferase (MPST) converts 3MP to hydrogen sulfide (H₂S). However, the regulatory mechanisms governing MPST and its impact on the brain remain largely unexplored. Our study reveals the neuroprotective role of endothelial MPST-generated H₂S, regulated by protein phosphatase 2A (PP2A). Bioinformatics analysis and RNA sequencing demonstrated that endothelial PP2A is associated with neurodegenerative disease pathways. Cerebral ischemic mice exhibited significant inactivation of endothelial PP2A, evidenced by the reduction of PP2Acα in the brain endothelium. Mice with endothelium-specific null PP2A (PP2A^EC-cKO) exhibited neuronal loss, cognitive dysfunction, and long-term potentiation deficits. Postnatal inactivation of endothelial PP2A also contributes to cognitive dysfunction and neuronal loss. However, regaining endothelial PP2A activity by overexpressing Ppp2ca rescued neuronal dysfunction. Mechanistically, PP2A deficiency is intricately linked to the MPST-H₂S signaling pathway. A robust reduction in endothelial MPST-dependent H₂S production followed PP2A deficiency. Exogenous H₂S treatment and AAV-mediated overexpression of MPST in brain endothelial cells significantly mitigated neuronal dysfunction in PP2A^EC-cKO mice. Furthermore, PP2A deficiency promotes an increase in calcium influx and calpain2 phosphorylation, subsequently leading to MPST degradation. The PP2A activator (FTY720) and MPST activator (3MP sodium) both remarkably restored endothelial MPST-dependent H₂S production, subsequently rescuing ischemia-induced neurological deficits. In conclusion, our study demonstrates that endothelial PP2A deficiency leads to MPST degradation by activating calpain2, thus damaging neuronal function.

Plant metabolites are important for plant development and human health. Plants of celery (Apiumgraveolens L.) with different-colored petioles have been formed in the course of long-term evolution. However, the composition, content distribution, and mechanisms of accumulation of metabolites in different-colored petioles remain elusive. Using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), 1159 metabolites, including 100 lipids, 72 organic acids and derivatives, 83 phenylpropanoids and polyketides, and several alkaloids and terpenoids, were quantified in four celery cultivars, each with a different petiole color. There were significant differences in the types and contents of metabolites in celery with different-colored petioles, with the most striking difference between green celery and purple celery, followed by white celery and green celery. Annotated analysis of metabolic pathways showed that the metabolites of the different-colored petioles were significantly enriched in biosynthetic pathways such as anthocyanin, flavonoid, and chlorophyll pathways, suggesting that these metabolic pathways may play a key role in determining petiole color in celery. The content of chlorophyll in green celery was significantly higher than that in other celery cultivars, yellow celery was rich in carotenoids, and the content of anthocyanin in purple celery was significantly higher than that in the other celery cultivars. The color of the celery petioles was significantly correlated with the content of related metabolites. Among the four celery cultivars, the metabolites of the anthocyanin biosynthesis pathway were enriched in purple celery. The results of quantitative real-time polymerase chain reaction (qRT-PCR) suggested that the differential expression of the chalcone synthase (CHS) gene in the anthocyanin biosynthesis pathway might affect the biosynthesis of anthocyanin in celery. In addition, HPLC analysis revealed that cyanidin is the main pigment in purple celery. This study explored the differences in the types and contents of metabolites in celery cultivars with different-colored petioles and identified key substances for color formation. The results provide a theoretical basis and technical support for genetic improvement of celery petiole color.
Anthocyanins ; Apium/metabolism* ; Chlorophyll/metabolism* ; Color ; Gene Expression Regulation, Plant ; Humans ; Metabolomics ; Plant Proteins/genetics* ; Tandem Mass Spectrometry

Objective:To evaluate the teaching effectiveness of independent experimental design from students' active learning behavior, and further provide the basis for advancing the reform of functional experimental teaching and teaching quality.Methods:In June 2019, 186 undergraduates (5-year-programme and 8-year-programme) of Xiangya School of Medicine were included in the teaching research. Self-administered questionnaires were applied to characterize students' active learning behavior in independent experimental design education. Spearman rank correlation analysis and Logistic regression analysis were used in the study. SPSS 23.0 was used for descriptive analysis of the data.Results:During the independent experimental design, 85.0%(158/186) of the students thought it was necessary and important to conduct independent experimental design education; 72.6%(135/186) of the students tentatively raised new scientific questions; 97.8%(182/186) of the students actively searched literature; 77.4%(144/186) of the students participated in reply positively. The value of correlation coefficient of actively learning behavior "tentatively raising new science questions" and teaching effectiveness "improving the ability of scientific thinking" was 0.81. And only 42.5%(79/186) of the students agreed that students needed to summarize after reporting.Conclusion:Independent experimental design education is welcomed and widely accepted by students, which has effectively improved the capacity for scientific research and innovation spirit of students. Whether students' active learning behavior can be fully mobilized in the education practice is closely related to the teaching effect. And the cultivation of leadership and leading consciousness still need to be improved.

Objective:To analyze the clinical characteristics, therapeutic options and risk factors of mortality in patients with carbapenem-resistant Acinetobacter baumannii (CRAB) bloodstream infection, and to provide evidence for clinical treatment option and prognosis evaluation of CRAB bloodstream infections. Methods:A retrospective study was carried out in 224 patients with confirmed diagnosis of CRAB bloodstream infection during the period from January 2012 to December 2017 in West China Hospital, Sichuan University. The patients were divided into the death group and the survival group according to the survival status 28 days after collecting blood samples. The clinical features and therapeutic options of antibacterial drugs were reviewed. Student′s t test was used for analyzing normally distributed data and Mann-Whitney U test for non-normal data.Chi-square test was used for categorical variables. Univariate and multivariate logistic analysis were used to analyze the risk factors of mortality associated with CRAB bloodstream infection. Results:Among 224 cases of CRAB bloodstream infection, 121 cases died (54.02%). These patients were mainly in intensive care unit (ICU) and hematology department. The common underlying diseases were severe acute pancreatitis and severe cardiovascular events. The interleukin (IL)-6 level (median (interquartile range)) in the death group (480.40 ng/L (1 432.95 ng/L)) was higher than that of the survival group (107.05 ng/L (263.08 ng/L)), the difference was statistically significant ( Z=4.526, P<0.01). The procalcitionin (PCT) levels in the death group and the survival group were 3.81 μg/L (17.26 μg/L) and 2.12 μg/L (12.74 μg/L), respectively, with no difference between the two groups ( P>0.05). The death rate of empirical treatment with a single or more non-active antimicrobial agents was 57.14% (64/112), that of monotherapy with active agent was 45.68% (37/81), and that of combination therapy with at least one active drug was 64.52% (20/31). The differences had no statistical significance ( P=0.130). The logistic regression analysis showed that the risk factors of mortality associated with CRAB bloodstream infection were renal dysfunction (odds ratio ( OR)=2.181, P=0.024) and multiple organ dysfunction syndrome (MODS; OR=20.376, P<0.01). Conclusions:The fatality rate of patients with CRAB bloodstream infection is high. These patients with renal dysfunction or MODS have poor prognosis. In addition to early effective antibacterial therapy, individual comprehensive treatment should be implemented in order to improve the curative effect.

OBJECTIVE: To evaluate the effects of the pestle needling therapy on the vibration perception threshold (VPT) and the quality of life (QOL) in the patients of high-risk diabetic foot. METHODS: A total of 132 patients of high risk diabetic foot were randomized into an observation group and a control group, 66 cases in each one. In the control group, the routine treatment and nursing for diabetes were applied. In the observation group, on the base of the treatment as the control group, the pestle needling therapy was used at , , , Zusanli (ST 36), Yongquan (KI 1), Sanyinjiao (SP 6) and Taixi (KI 3). The intervention with the pestle needling therapy was given once every day, consecutively for 4 weeks. The follow-up visit was conducted in 3 months and 6 months after the intervention. Vibration perception threshold (VPT) was observed before and after intervention in the two groups. Separately, before intervention, at the end of intervention, 3 months after intervention and 6 months after intervention, the type 2 diabetes mellitus quality life scale (DMQLS), the revised edition was adopted to compare the QOL between the two groups. The effects were evaluated in the two groups. RESULTS: After intervention, the VPT and QOL were all improved in the two groups (<0.01, <0.05). In the observation group, after intervention, VPT at the first toes and the dorsum of the feet was lower than the control group at the same time point (all <0.01). In the observation group, the total scores and the score of each dimension item in the DMQLS of the revised edition were all lower than the control group correspondently at the end of intervention, 3 months after intervention and 6 months after intervention (<0.01, <0.05). The repeated ANOVA results showed that the differences were significant in the total scores and the score of each dimension in the aspects of time, inter-group and interaction effect (<0.01, <0.05). The total scores and score of each dimension item in the DMQLS were lower than those the earlier time point (<0.01, <0.05). The total effective rate was 100.0% in the observation group, which was superior to 95.1% (59/62) in the control group (<0.05). CONCLUSION The pestle needling therapy reduces the foot VPT and improves the sensory nerve function of the foot and the QOL in the patients of high-risk diabetic foot.
Acupuncture Therapy ; Diabetes Mellitus, Type 2 ; Diabetic Foot ; therapy ; Humans ; Perception ; Quality of Life ; Vibration

OBJECTIVETo explore the clinical efficacy and safety of acupuncture and moxibustion for copracrasia.

METHODSBy prospective live randomized controlled trial, 40 cases with copracrasia were randomly divided into an acupuncture-moxibustion group and a medication group, 20 cases in each one. In the acupuncture-moxibustion group, acupuncture was applied at Ciliao (BL 32), Changqiang (GV 1) and Tianshu (ST 25) and mild moxibustion was used at Qihai (CV 6). Treatment was given for 12 weeks and 32 times, 3 times a week in the front 8 weeks, 2 times a week in the latter 4 weeks. In the medication group, conventional symptomatic treatment, support therapy, and complications preventing and treating were adopted for 12 weeks. Anal incontinence score (Vaizey incontinence score), effective rate and self-rating score for satisfaction were observed before and after treatment and in the follow-up period.

RESULTSAfter 12 weeks' treatment in the two groups, Vaizey incontinence' scores were both decreased (both P<0. 05), and after treatment and in the follow-up period the scores in the acupuncture-moxibustion group were lower than those in the corresponding period in the medication group (both P< 0. 05). The effective rate of the acupuncture-moxibustion group was 80. 0% (16/20), which was statistically different from 50. 0% (10/20) in the medication group (P<0. 05). The effective rate in the follow-up period of the acupuncture-moxibustion group was 90. 0% (18/20) and it was not statistically different from 80. 0% (16/20) in the medication group (P>0. 05). The self-rating scores for satisfaction in the acupuncture-moxibustion group were superior to those in the medication group after treatment and in the follow-up period (both P< 0. 05).

CONCLUSIONAcupuncture and moxibustion could improve copracrasia and the acupuncture-moxibustion rules and characteristics for the disorder should be paid attention to in the further research.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Defecation ; Fecal Incontinence ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

Objective To evaluate the feasibility and safety of vaginal enlarged amputation of cervix to treat patients with cervical cancer of stage Ⅰ a1 and cervical intraepithelial neoplasia grade Ⅲ(CIN Ⅲ)who were unfit for conization surgery.Methods From July 2002 to May 2007,patients with cervical cancer at stage Ⅰ a1,diagnosed by pathology after loop electrosurgical excision procedure(LEEP),large area CIN Ⅲ(the area of lesion≥3/4 on colposcopy),CIN Ⅲ coexisted with vaginal intraepithelial neoplasia (VAIN)in the superior segment of vagina,CIN Ⅱ-Ⅲ recurrence or with residual lesion,positive margin after conization of cervix,who wanted to preserve fertility and(or)corpus uteri were selected to receive vaginal enlarged amputation of cervix.Results Forty-eight eases including 5 with cervical cancer in stage Ⅰ a1,38 with large area CIN Ⅲ(9 with gland involvement),2 with residual lesion and 2 with positive margin after LEEP,1 recurrence after cold knife conization,received the procedure successfully.The median age was 34 years(range 27-40),median operation time was 60 minutes(range 30-100),median blood loss was 40 ml(range 5-300),and median hospital stay was 10 days(range 7-17).After follow-up 1-39 months,no patient had postoperative complications and recurrence,and all patients resumed normal menstrual cycle and sexual life.Condusion Vaginal enlarged amputation of cervix appears to be a safe and feasible procedure for patients with cervical cancer at stage Ⅰ a1 and CIN Ⅲ who are unfit for conization surgery.