Six compounds were isolated from the ethyl acetate fraction of Citri Sarcodactylis Fructus by using various column chromatographic methods, such as MCI Gel CHP-20, ODS, Sephadex LH-20, silica gel and semipreparative HPLC. Their structures were identified to be citrusin G (1), citrusin H (2), citrusin E (3), syringin (4), coniferin (5), methylconiferin (6) by NMR, HR-ESI-MS, UV, IR spectra. Compounds 1 and 2 were new secondary metabolism products and 3-6 were obtained from the titled material for the first time. Compound 1 showed anti-renal fibrosis activity in TGF-β1-induced kidney proximal tubular cells.

OBJECTIVE: To determine the role of Tripterygium wilfordii multiglycoside (TGW) in the treatment of psoriatic dermatitis from a cellular immunological perspective. METHODS: Mouse models of psoriatic dermatitis were established by imiquimod (IMQ). Twelve male BALB/c mice were assigned to IMQ or IMQ₊TGW groups according to a random number table. Histopathological changes in vivo were assessed by hematoxylin and eosin staining. Ratios of immune cells and cytokines in mice, as well as PAM212 cell proliferation in vitro were assessed by flow cytometry. Pro-inflammatory cytokine expression was determined using reverse transcription quantitative polymerase chain reaction. RESULTS: TGW significantly ameliorated the severity of IMQ-induced psoriasis-like mouse skin lesions and restrained the activation of CD45⁺ cells, neutrophils and T lymphocytes (all P<0.01). Moreover, TGW significantly attenuated keratinocytes (KCs) proliferation and downregulated the mRNA levels of inflammatory cytokines including interleukin (IL)-17A, IL-23, tumor necrosis factor α, and chemokine (C-X-C motif) ligand 1 (P<0.01 or P<0.05). Furthermore, it reduced the number of γ δ T17 cells in skin lesion of mice and draining lymph nodes (P<0.01). CONCLUSIONS TGW improved psoriasis-like inflammation by inhibiting KCs proliferation, as well as the associated immune cells and cytokine expression. It inhibited IL-17 secretion from γ δ T cells, which improved the immune-inflammatory microenvironment of psoriasis.
Male ; Animals ; Mice ; Tripterygium ; Psoriasis/drug therapy* ; Keratinocytes ; Skin Diseases/metabolism* ; Cytokines/metabolism* ; Imiquimod/metabolism* ; Dermatitis/pathology* ; Disease Models, Animal ; Mice, Inbred BALB C ; Skin/metabolism*

Objective:To investigate the predictive value of controlling nutritional status (CONUT) score in the prognosis of patients with advanced diffuse large B-cell lymphoma (DLBCL).Methods:A retrospective case series study was performed. The clinical data of 654 patients newly diagnosed with advanced DLBCL diagnosed in 7 medical centers in Huaihai Lymphoma Working Group from October 2009 to January 2022 were retrospectively collected. All the patients received rituximab-based immune chemotherapy regimens. The patients were randomly assigned to the training set (458 cases) and the validation set (196 cases) in a 7:3 ratio. The clinicopathological data of patients were collected, and the CONUT score was calculated based on albumin, lymphocyte count, and total cholesterol. The optimal critical value of CONUT scote was determined by using MaxStat method. Kaplan-Meier method was used to draw survival curves; Cox proportional hazards model was used to make univariate analysis and multivariate analysis on the factors influencing overall survival (OS). The efficacy of CONUT score in combination with the International prognostic index (IPI) and an enhanced IPI (NCCN-IPI) in predicting OS was evaluated by using receiver operating characteristic (ROC) curves.Results:The median follow-up time of 654 patients was 38.1 months (95% CI: 35.3 months- 40.9 months), and the 5-year OS rate was 49.2%. According to the MaxStat method, the optimal critical value for CONUT score was determined to be 6 points. All the patients were classified into the normal nutritional status group (CONUT score ≤ 6 points, 489 cases) and the poor nutritional status group (CONUT score > 6 points, 165 cases). The results of the multivariate analysis showed that CONUT score > 6 points, male, lactate dehydrogenase >240 U/L, high white blood cell count, low hemoglobin level and age > 60 years were independent risk factors for OS of patients with advanced DLBCL (all P < 0.05). Patients in the poor nutritional status group (CONUT score > 6 points) had worse OS compared with that in the normal nutritional status group in the overall cohort of advanced DLBCL. Subgroup analysis revealed that among patients with Eastern Cooperative Oncology Group-performance status (ECOG PS) score < 2 points, IPI low-intermediate risk, IPI intermediate-high risk, NCCN-IPI low-intermediate risk, and NCCN-IPI intermediate-high risk, the patients in the poor nutritional status group (CONUT score > 6 points) had worse OS compared with that in the normal nutritional status group (CONUT score ≤ 6 points) (all P < 0.05). Conclusions:CONUT score has a certain value in the assessment of the prognosis of patients with advanced DLBCL, and its predictive efficacy is further improved when combined with IPI and NCCN-IPI.

Objective To investigate the expression of PLCD3 mRNA in the synovium of osteoarthritis(OA)and its relationship with immune cell infiltration.Methods Based on the differentially expressed genes of OA found in the previous study,the expression of phospholipase Cδ3(PLCD3)mRNA was detected by col-lecting synovial samples from OA group and control group.CIBERSORT algorithm was used to analyze the infiltration pattern of immune cells in OA group and control group,and the correlation between PLCD3 and infiltrating immune cells was further analyzed.Results Compared with the control group,the relative expres-sion level of PLCD3 mRNA was significantly increased in synovial samples of OA group(P<0.05).The pro-portions of B cells naive,NK cells activated,M2 macrophages and mast cells activated in synovial tissues of OA group were relatively high(P<0.05).PLCD3 was positively correlated with the proportion of these four immune cells(P<0.05).Conclusion PLCD3 may be a key biomarker for the diagnosis of OA,which may be involved in the pathogenesis of OA by interacting with infiltrating immune cells.

Objective:To analyze and compare the differences of expression profiles between RPL and normal adipose tissue by transcriptome sequencing(RNA-Seq),then identify the key genes related to prognosis and explore their potential mechanisms.Methods:Tumor tissues and normal adipose tissues of patients with RPL were collected for RNA-Seq,and then the differentially ex-pressed genes were analyzed by GO and KEGG enrichment analysis.Based on TCGA,the high-risk genes related to prognosis were screened and verified by Kaplan-Meier curve and receiver operat-ing characteristic(ROC)curve.Results:Compared with normal adipose tissue,279 differentially expressed genes were simultaneously up-regulated in RPL tissues,which were mainly enriched in immune response and PPAR signaling pathway.Combined with TCGA,7 stable prognostic high risk genes were identified and the overall survival rate of the high risk group was significantly lower than that of the low risk group(P＜0.05).Conclusion:KCNQ5,RBPJ and some other genes may be re-lated to the poor prognosis of RPL patients.The analysis of the mechanism of these genes in RPL is expected to provide new evidence for the formulation of diagnosis and treatment strategies for RPL patients.

Objective:To study the osseointegration and new bone formation of titanium implants treated by sandblasting acid etching combined with plasma spraying tantalum coating in rats.Methods:After polishing and cleaning of the pure titanium specimens(SLA),the tantalum coating was prepared by plasma spraying after sandblasting and acid etching(SLA-Ta).The surface feature of the samples was observed by SEM,EDS and contact angle meter.The samples of SLA and SLA-Ta were respectively implanted into femur condyle at each side of 18 SD rats(SLA group and SLA-Ta group),and the rats were double-fluorescent labeled after surgery.The rats were sacrificed 4 and 12 weeks after implantation respectively(n=9),the samples of implants and surrounding tissue were examined by mi-cro-CT scanning,fluorescence microscopy and histological staining,the new bone formation and osseointegration around the implant were analyzed.Results:Micro-nano Porous structureed tantalum coating was successfully prepared on the surface of SLA titanium.In vivo experiments showed that 4 and 12 weeks after implantation,the amount of new bone and osseointegration around the implant in the SLA-Ta group were significantly higher than those in the SLA group.Conclusion:The micro-nano multi-level structure formed by SLA combined with plasma sprayed tantalum can promote new bone formation,accelerate mineralization deposition,and improve the osseoin-tegration capacity of the implant.

Objective To investigate the role and underlying mechanisms of WD repeat domain 82(WDR82)protein in the pathogenesis and progression of glioma.Methods We analyzed the expression level of WDR82 in glioma tissues using GEPIA and UALCAN databases and further assessed WDR82 protein expression in glioma and adjacent normal tissues through immunohistochemical staining.The correlation between WDR82 expression and the prognosis of glioma patient was evaluated using Kaplan-Meier plotter.Experiments were conducted on A172 and U251 cell lines,which were categorized into four groups:control group(transfected with 3 μg pcDNA3),shR-control group(transfected with 3 μg pSilencer 2.1-U6),pWDR82 group(transfected with 3 μg pWDR82),and shR-WDR82 group(transfected with 3 μg shR-WDR82).Post-transfection,we confirmed transfection efficiency at 48 hours using qRT-PCR and measured cell viability at the same time point with CCK-8 assay.Clone formation assay was employed to assess cell proliferation capacity after 14 days of transfection,while flow cytometry was utilized to analyze cell apoptosis after 48 hours of transfection.Additionally,Western blotting was conducted to determine the expression levels of proteins related to proliferation and Akt/mTOR signaling pathway after 48 hours of transfection.Finally,the effect of WDR82 on tumor growth in NOD-SCID mice was investigated using tumor carrying experiment in vivo.Results Analysis of WDR82 expression in glioma tissues using GEPIA and UALCAN databases,along with immunohistochemical staining,revealed significantly higher expression levels compared to normal and paracancerous tissues(P＜0.05).Additionally,WDR82 expression was not associated with gender or age of patients(P＞0.05).Kaplan-Meier plotter analysis indicated that elevated WDR82 expression correlated with a poor prognosis in glioma patients(log-rank P=0.029).Overexpression of WDR82 notably enhanced the proliferation and inhibited the apoptosis of A172 and U251 cells,and also significantly upregulated the expression of MKI67,BCL2,CCND1,p-Akt and p-mTOR in A172 and U251 cells(P＜0.05).Conversely,WDR82 knockdown had the opposite effects,inhibiting cell proliferation,increasing apoptosis and downregulating the expression of MKI67,BCL2,CCND1,p-Akt and p-mTOR(P＜0.05).WDR82 knockdown in U251 cells significantly inhibited tumor growth in NOD-SCID mice(P＜0.05).Conclusion High expression of WDR82 promotes the proliferation of glioma cells and the growth of tumors in vivo by regulating the AKT/mTOR signaling pathway.

To study the carriage status of drug susceptibility, clonal complex groups, serotypes, surface proteins and virulence genes of Streptococcus agalactiae from respiratory specimen sources. A total of 35 strains of S.agalactiae meeting the criteria were collected from 3 hospitals in 2 locations, Tangshan and Jinan. The age span of the patients was 3 days-92 years, and the percentage of elderly patients&ge;60 years was 71.5%.The susceptibility to 9 antimicrobial drugs was measured and analyzed using the micro broth dilution method. The strains were 100.0% sensitive to penicillin, linezolid, vancomycin, and ceftriaxone; However, it exhibits high resistance rates to erythromycin, clindamycin and levofloxacin, at 97.1%, 85.7% and 82.9% respectively; and the resistance rates to tetracycline and chloramphenicol were 34.3% and 14.2%, respectively. Genome sequence determination and analysis showed that 16 resistance genes were detected in 35 strains, among which: macrolide and lincosamide resistance genes were mainly ermB, with a carrying rate of 74.2%; tetracycline resistance genes were mainly tetM, with a carrying rate of 25.7%; in addition, the mutation rates of the quinolone resistance determinants gyrA and parC were 88.5% and 85.7%, respectively. 35 strains belonged to 6 ST types and 4 clonal groups, with CC10/ST10 as the main one, accounting for 62.8%; they contained 4 serotypes of Ⅰb, Ⅱ, Ⅲ, and Ⅴ, as well as 1 untyped strain, with serotype Ⅰb as the main one, accounting for 65.7%. The strains carried three pilus types, PI1+PI2a, PI2a and PI2b types, respectively, and detected five surface proteins, alpha, alp1, rib, srr, and r df_0594, and seven virulence factors, cba, cfb, cylE, fbsA, hylB, l mb, and pavA. Overall, S.agalactiae isolated from respiratory tract specimens is predominantly sourced from elderly patients, with CC10 strains being most prevalent. These strains harbor multiple drug-resistant and virulence genes, demonstrating elevated resistance rates to macrolides, lincosamides, and quinolones. This emphasizes the necessity for vigilant attention to the health threat posed by S. agalactiae from respiratory tract speciments of elderly patients.

To study the carriage status of drug susceptibility, clonal complex groups, serotypes, surface proteins and virulence genes of Streptococcus agalactiae from respiratory specimen sources. A total of 35 strains of S.agalactiae meeting the criteria were collected from 3 hospitals in 2 locations, Tangshan and Jinan. The age span of the patients was 3 days-92 years, and the percentage of elderly patients&ge;60 years was 71.5%.The susceptibility to 9 antimicrobial drugs was measured and analyzed using the micro broth dilution method. The strains were 100.0% sensitive to penicillin, linezolid, vancomycin, and ceftriaxone; However, it exhibits high resistance rates to erythromycin, clindamycin and levofloxacin, at 97.1%, 85.7% and 82.9% respectively; and the resistance rates to tetracycline and chloramphenicol were 34.3% and 14.2%, respectively. Genome sequence determination and analysis showed that 16 resistance genes were detected in 35 strains, among which: macrolide and lincosamide resistance genes were mainly ermB, with a carrying rate of 74.2%; tetracycline resistance genes were mainly tetM, with a carrying rate of 25.7%; in addition, the mutation rates of the quinolone resistance determinants gyrA and parC were 88.5% and 85.7%, respectively. 35 strains belonged to 6 ST types and 4 clonal groups, with CC10/ST10 as the main one, accounting for 62.8%; they contained 4 serotypes of Ⅰb, Ⅱ, Ⅲ, and Ⅴ, as well as 1 untyped strain, with serotype Ⅰb as the main one, accounting for 65.7%. The strains carried three pilus types, PI1+PI2a, PI2a and PI2b types, respectively, and detected five surface proteins, alpha, alp1, rib, srr, and r df_0594, and seven virulence factors, cba, cfb, cylE, fbsA, hylB, l mb, and pavA. Overall, S.agalactiae isolated from respiratory tract specimens is predominantly sourced from elderly patients, with CC10 strains being most prevalent. These strains harbor multiple drug-resistant and virulence genes, demonstrating elevated resistance rates to macrolides, lincosamides, and quinolones. This emphasizes the necessity for vigilant attention to the health threat posed by S. agalactiae from respiratory tract speciments of elderly patients.

Objective:To investigate the alterations in brain dynamic functional network connectivity (dFNC) and their significance in Parkinson's disease (PD) patients with postural instability/gait difficulty (PIGD).Methods:Ninety PD patients admitted to Department of Neurology, First Affiliated Hospital of Nanjing Medical University from May 2016 to August 2019 were recruited, and 54 healthy controls matched with gender and age were chosen; their clinical data and resting-state functional MRI (rs-fMRI) data were collected. PD patients were divided into PD with PIGD (PD-PIGD) group ( n=49) and PD without PIGD (PD-non-PIGD) group ( n=41) according to Unified Parkinson's Disease Rating Scale (UPDRS) scores. Independent component analysis (ICA), sliding window method and k-means clustering were used to analyze the dFNC and compare among groups. Correlations of dFNC alterations with clinical scales were verified by partial correlation analysis. Results:Four repeated recurring functional connectivity states were identified, and PD-PIGD patients had high frequency in state 3 (44%) and state 2 (23%) of the low dFNC. In terms of dFNC time attributes, PD-PIGD patients had longer mean dwell time in state 3 than PD-non-PIGD patients and had lower number of transitions in state 3 than PD-non-PIGD patients and healthy controls, with significant differences ( P<0.05); PD-PIGD patients had significantly higher fractional windows and statistically longer mean dwell time in state 2 than healthy controls ( P<0.05). In terms of dFNC strengths, compared with healthy controls, PD-PIGD patients showed significantly decreased functional connectivity within default mode network (DMN, between medial superior frontal gyrus and precuneus) and auditory network (AN, between superior temporal gyrus and middle temporal gyrus), but significantly increased functional connectivity between sensorimotor network (SMN, supplementary motor area) and DMN (precuneus) in state 2 ( P<0.05, false discovery rate [FDR]-corrected). Partial correlation analysis indicated positive correlation between mean dwell time in state 3 and PIGD scores in PD-PIGD patients ( r=0.450, P=0.039). Conclusion:PD-PIGD patients exhibit specific dFNC, mainly characterized by low connectivity of the brain functional network and prolonged dwell time; local functional network domains often separate between DMN, AN and SMN networks and within the networks.