OBJECTIVE: To investigate the expression and physiological significance of the ferroptosis marker 4-hydroxynonenal (4-HNE) in myofibroblasts induced by transforming growth factor-β1 (TGF-β1), providing theoretical evidence for its potential role in the diagnosis and treatment of fibrosis in systemic sclerosis (SSc). METHODS: Mouse embryonic fibroblasts (NIH3t3) were cultured and divided into two groups after 12 h of starvation: the control group (cultured in 1% serum-containing medium) and the TGF-β1 group (cultured in 10 μg/L TGF-β1 with 1% serum-containing medium). Cell morphology changes in both groups were observed under a microscope. To confirm successful establishment of the SSc cell model, fibrosis markers were analyzed using reverse transcription quantitative real-time PCR (RT-qPCR) and Western blot. Next, flow cytometry was employed to assess the intracellular levels of reactive oxygen species (ROS) in both groups. Finally, Western blot and immunofluorescence staining were used to measure the expression of 4-HNE in the TGF-β1-treated cells. RESULTS: Microscopic observations revealed that TGF-β1 treatment caused the NIH3t3 cells to transition from a typical spindle shape to a flat, polygonal shape with multiple protrusions, indicating fibroblast activation. The RT-qPCR and Western blot analyses showed that the expression of the fibrosis marker Vimentin was significantly upregulated in the TGF-β1 group compared with the control group (P < 0.01), confirming that TGF-β1 effectively promoted fibrosis-related gene and protein expression. Flow cytometry results indicated that TGF-β1 significantly elevated intracellular ROS levels, suggesting the induction of oxidative stress. Furthermore, both Western blot and immuno-fluorescence staining demonstrated a significant increase in 4-HNE expression in the TGF-β1-treated cells (immunofluorescence intensity P < 0.05). CONCLUSION TGF-β1 promotes fibroblast activation and fibrosis while inducing ROS production, leading to a marked increase in 4-HNE expression. Given the role of 4-HNE as a marker of lipid peroxidation and its elevated levels in the SSc cell model, this study suggests that 4-HNE could serve as a potential biomarker for fibrosis in SSc. The findings highlight the importance of investigating the mechanisms of 4-HNE in fibrosis and suggest that targeting this pathway could offer new therapeutic opportunities for treating SSc.
Animals ; Mice ; Scleroderma, Systemic/pathology* ; Aldehydes/pharmacology* ; Transforming Growth Factor beta1/metabolism* ; NIH 3T3 Cells ; Ferroptosis ; Reactive Oxygen Species/metabolism* ; Fibrosis ; Fibroblasts/metabolism* ; Biomarkers/metabolism* ; Myofibroblasts/metabolism*

[Abstract] Objective To explore the effect of serine protease inhibitor Kazal-type 1(SPINK1) on the proliferation of hepatocellular carcinoma cells RH-35 and its underling molecular mechanism. Methods Spink1 gene expression in liver cancer and rat liver cancer models were analyzed by Gene Expression Omnibus (GEO) data, RH-35 cells were treated with rrSPINK1 protein, the effect of rrSPINK1 on the proliferation and apoptosis of RH-35 cells was explored by MTT, 2’-deoxy-5-ethynyluridine(EdU) and flow cytometry, the molecular mechanism of SPINK1 regulating liver cancer were detected by Real-time PCR and Western blotting. Results The results showed that Spink1 gene was over-expressed significantly in liver cancer and rat liver cancer models, rrSPINK1-treated RH-35 cells showed increased viability, EdUpositive cell rate, and the proportion of cells in S phase and G

Abstract: Objective To analyze the antimicrobial resistance rate and risk factors of multi drug resistant organisms (MDRO) in bloodstream infection for rational treatment. Methods　A total of 696 cases of bloodstream infections of Staphylococcus aureus, Enterococcus, Enterobacteriaceae (excluding Salmonella and Shigella), Pseudomonas aeruginosa and Acinetobacter in our hospital from 2017 to 2021 were selected, and 711 pathogenic strains were isolated from their whole blood samples. The antimicrobial resistance rates of various multi drug resistant strains were analyzed and the risk factors of MDRO infection were analyzed. Results　696 non repeated cases were screened out from 13 187 whole blood culture specimens, with a positive rate of 5.3%, and a total of 711 blood influenza pathogens were detected, among them, 350 strains of MDRO were detected with a detection rate of 49.23% (350/711). Among the pathogenic bacteria of bloodstream infection, Escherichia coli was the most, with 277 strains accounting for 38.96% (277/711), of which 201 strains were MDRO, accounting for 57.43% (201/350); followed by Klebsiella pneumoniae and Staphylococcus aureus, with 155 strains accounting for 21.80% (155/711) and 89 strains accounting for 12.52% (89/711), among which 43 strains of Klebsiella pneumoniae MDRO accounted for 12.29% (43/350) and 38 strains of Staphylococcus aureus MDRO accounted for 10.86% (38/350). The change trend of the three pathogens during 2017-2021 was not obvious. The drug sensitivity test showed that Escherichia coli and Klebsiella pneumoniae were highly resistant to cephalosporins and fluoroquinolones, and the drug resistance rate of aminoglycosides was relatively low. They had almost no resistance to cephalosporins and carbapenems. Staphylococcus aureus has a high resistance rate to lincomycin and macrolides, but no resistance to oxazolidinone, glycopeptides and glycylcyclins. There were 350 cases of MDRO infection and 361 cases of non MDRO infection. Univariate analysis showed that the age, sex, cardiovascular and cerebrovascular history, renal insufficiency, lung disease, hypoalbuminemia, hepatobiliary disease, electrolyte disorder and anemia of the patients had no statistical significance in MDRO infection (P>0.05); diabetes, urinary tract infection, surgical operation and burn were the influencing factors of MDRO (P<0.05). According to logistic analysis, diabetes, urinary tract infection, surgical operation and burn were the risk factors of MDRO infection (P<0.05). Conclusion　The infection of MDRO in patients with bloodstream infection is serious, and early prevention and control should be paid attention to, and the principle of graded use of antibiotics should be strictly observed, and the rational application should be carried out to actively and effectively control the production of MDRO.

OBJECTIVES: To explore the optimal maintenance dose of caffeine citrate for preterm infants requiring assisted ventilation and caffeine citrate treatment. METHODS: A retrospective analysis was performed on the medical data of 566 preterm infants (gestational age ≤34 weeks) who were treated and required assisted ventilation and caffeine citrate treatment in the neonatal intensive care unit of 30 tertiary hospitals in Jiangsu Province of China between January 1 and December 31, 2019. The 405 preterm infants receiving high-dose (10 mg/kg per day) caffeine citrate after a loading dose of 20 mg/kg within 24 hours after birth were enrolled as the high-dose group. The 161 preterm infants receiving low-dose (5 mg/kg per day) caffeine citrate were enrolled as the low-dose group. RESULTS: Compared with the low-dose group, the high-dose group had significant reductions in the need for high-concentration oxygen during assisted ventilation (P=0.044), the duration of oxygen inhalation after weaning from noninvasive ventilation (P<0.01), total oxygen inhalation time during hospitalization (P<0.01), the proportion of preterm infants requiring noninvasive ventilation again (P<0.01), the rate of use of pulmonary surfactant and budesonide (P<0.05), and the incidence rates of apnea and bronchopulmonary dysplasia (P<0.01), but the high-dose group had a significantly increased incidence rate of feeding intolerance (P=0.032). There were no significant differences between the two groups in the body weight change, the incidence rates of retinopathy of prematurity, intraventricular hemorrhage or necrotizing enterocolitis, the mortality rate, and the duration of caffeine use (P>0.05). CONCLUSIONS This pilot multicenter study shows that the high maintenance dose (10 mg/kg per day) is generally beneficial to preterm infants in China and does not increase the incidence rate of common adverse reactions. For the risk of feeding intolerance, further research is needed to eliminate the interference of confounding factors as far as possible.
Caffeine/therapeutic use* ; Citrates ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Respiration, Artificial ; Retrospective Studies

Hypoxic pulmonary hypertension (HPH) is characterized by hypoxic pulmonary vasoconstriction (H P V) and hypoxic pulmonary vascular remodeling(HPVR). Previous studies have shown that intracellular Ca

Objective:To establish a high performance liquid chromatography (HPLC) method for determination of four active constituents, namely MSTG-A,MSTG-B,gaultherin and chlorogenic acid in the anti-inflammatory and analgesic active fraction (ARF) of the ethnic medicine Gaultheria leucocarpa var. yunnanensis, in order to provide a methodological basis for the in-depth study and quality control of G. leucocarpa var. yunnanensis,and lay a foundation for later preparation and clinical application. Method:The determination was performed on COSMOSIL 5C₁₈-PAQ (4.6 mm×250 mm,5 μm) column with methanol-0.2% glacial acetic acid (gradient elution) as the mobile phase at a flow rate of 1 mL·min^-1. The column temperature was 25℃. The detection wavelength was set at 294 nm. Result:The linear range of MSTG-B,MSTG-A,gaultherin and chlorogenic acid were 0.014 06-0.450 00,0.007 81-0.250 00,0.003 13-0.100 00,0.000 94-0.030 00 g·L^-1 (r ≥ 0.999 7),respectively,with a good precision,repeatability and stability. And the average recoveries were 100.81%,98.99%,96.12% and 102.56%,respectively. RSDs were 1.4%,0.7%,0.7%,2.4%,respectively. The contents of MSTG-B,MSTG-A,gaultherin and chlorogenic acid in ARF fraction of G. leucocarpa var. yunnanensis were 23.608,41.973,8.282,2.673 mg·g^-1,respectively. Conclusion:The established method was simple and accurate, with a high repeatability. It can be used for determination of four active constituents in ARF fraction of G. leucocarpa var. yunnanensis,so as to provide a reference for the in-depth research,quality control and comprehensive evaluation of G. leucocarpa var. yunnanensis and lay a solid foundation for preparation and clinical application.

OBJECTIVETo summarize the clinical characteristics of peripheral blood, immune phenotypes, fusion genes and cytogenetics of patients with t(8;21) acute myeloid leukemia(AML) through the retrospective analysis of 586 patients with t(8;21) AML from 15 blood disease research centers in Northern area of China.

METHODSThe factors affecting prognosis of patients with t(8;21) AML were investigated by using univariate and multivariate COX regression.

RESULTSThe immune type of t(8;21) AML patients was mainly with HLA-DR, CD117, CD34, MPO, CD38, CD13and CD33(>95%), part of them with CD19and CD56; the most common accompanied mutation of t(8;21) AML patients was C-KIT mutation (37.8%); in addition to t(8;21) ectopic, the most common chromosomal abnormality was sex chromosome deletions (38.9%). The univariate analysis revealed a significant survival superiority of OS and PFS in t(8;21) AML patients of WBC≤3.5×10/L without C-KIT mutation, the newly diagnosed ones achieved HSCT(P<0.05), only survival superiority on OS in t(8;21) AML patients with extramedullary infiltration and CD19 positive; the results of multivariate analysis showed a significant survival superiority on OS and PFS in t(8;21) AML patients with WBC≤3.5×10/L(P<0.05).

CONCLUSIONThe clinical features of t(8;21) AML patients in China are similar to those in other countries, WBC≤3.5×10/L is a good prognostic factor while the C-KIT mutation is a poor one in t(8;21) AML patients.

To investigate the effect of piperine on the disorder of glucose metabolism in the cell model with insulin resistance (IR) and explore the molecules mechanism on intervening the upstream target of AMPK signaling pathway. The insulin resistance models in HepG2 cells were established by fat emulsion stimulation. Then glucose consumption in culture supernatant was detected by GOD-POD method. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of leptin(LEP) and adiponectin(APN) in culture supernatant; Real-time quantitative PCR was used to assess the mRNA expression of APN and LEP; and the protein expression levels of LepR, AdipoR1, AdipoR2 and the activation of AMPK signaling pathway were detected by Western blot analysis. The results showed that piperine, rosiglitazone and AMPK agonist AICAR could significantly elevate the glucose consumption in insulin resistance cell models, enhance the level of APN, promote APN mRNA transcripts and increase the protein expression of Adipo receptor. Meanwhile,AMPKα mRNA and р-AMPKα protein expressions were also increased in piperine treated cells, but both LEP mRNA expression and LepR protein expressions were decreased in piperine treated group. The results indicated that piperine could significantly ameliorate the glucose metabolism disorder in insulin resistance cell models through regulating upstream molecules (APN and LEP) of AMPK signaling pathway, and thus activate the AMPK signaling pathway.

Objective To establish the three-dimensional finite element model of human lower cervical spine C3-7 motion segments after anterior cervical corpectomy and fusion (ACCF) surgery with titanium mesh and bone graft,and to analyze the stability of cervical spine and stress distribution of internal fixation devices after ACCF surgery.Methods The finite element model of cervical spine C3-7 segments after ACCF of C5 segment with titanium mesh,bone graft,plate and screw fixation was established,and C3-7 segment intact model of cervical vertebra was also constructed.The torque moment of 0.5,1.0,1.5,2.0 N · m was applied to the ACCF surgery model.The ROM,maximum stress in facet joint and stress distributions on internal fixation devices under flexion,extension,lateral bending and axial rotation movement were analyzed.Results ROM of reconstructed C5 segment increased with the torque moment increasing after ACCF surgery.In the case of 1.0 N · m tomue moment and 50 N preload,the ROM of reconstructed C5,C3-4,C6-7 and C3-7 segment was reduced by 81％,62％,58％ and 80％ compared with the intact model.The maximum stress in facet joint of reconstructed C5 segment reduced and the stress in adjacent segments significantly increased.The stress of titanium mesh was mainly distributed on the compression side of movement,and high stress was located in the roots of screws.Conclusions ACCF surgery can promote the stability of cewical spine,decrease the stress in facet joint of operation segment,and has better treatment effect on easing compression from spinal cord caused by cervical spondylotic myelopathy.The research results will provide some theoretical basis for clinical application of ACCF surgery.

Objective To establish the three-dimensional finite element model of human lower cervical spine C3-7 motion segments after anterior cervical corpectomy and fusion (ACCF) surgery with titanium mesh and bone graft, and to analyze the stability of cervical spine and stress distribution of internal fixation devices after ACCF surgery. Methods The finite element model of cervical spine C3-7 segments after ACCF of C5 segment with titanium mesh, bone graft, plate and screw fixation was established，and C3-7 segment intact model of cervical vertebra was also constructed. The torque moment of 0.5, 1.0, 1.5, 2.0 N﹒m was applied to the ACCF surgery model. The ROM, maximum stress in facet joint and stress distributions on internal fixation devices under flexion, extension, lateral bending and torsion movement were analyzed. Results ROM of reconstructed C5 segment increased with the torque moment increasing after ACCF surgery. In the case of 1.0 N﹒m torque moment and 50 N preload, the ROM of reconstructed C5, C3-4, C6-7 and C3-7 segment was reduced by 81%, 62%, 58% and 80% compared with the intact model. The maximum stress in facet joint of reconstructed C5 segment reduced and the stress in adjacent segments significantly increased. The stress of titanium mesh was mainly distributed on the compression side of movement, and high stress was located in the roots of screws. Conclusions ACCF surgery can promote the stability of cervical spine, decrease the stress in facet joint of operation segment, and has better treatment effect on easing compression from spinal cord caused by cervical spondylotic myelopathy. The research results will provide some theoretical basis for clinical application of ACCF.