Objective To investigate the mechanism by which fragile X mental retardation protein(FMRP)regulates ferroptosis evasion in colorectal cancer(CRC)cells.Methods We examined FMRP expression levels in CRC cell lines using RT-qPCR and Western blotting and analyzed the biological functions and signaling pathways involved in FMRP-mediated regulation of CRC progression using the TCGA database.A lentiviral FMRP overexpression vector(Lv-FMRP)and 3 knockdown vectors(siFMRP-1,siFMRP-2,and siFMRP-3)were constructed,and their effects on proliferation of HCT116 cells were examined using CCK8 assay and plate clone formation assay;the changes in cell ferroptosis level was determined using MDA/ROS/GSH/Fe2+kits,mitochondrial membrane potential changes were detected using JC-1 fluorescence staining,and the expressions of proteins associated with ferroptosis and the RAS/MAPK signaling pathway were detected using Western blotting.The subcutaneous tumorigenic potential of the transfected cells was evaluated in nude mice.Results Compared with normal colonic mucosal epithelial NCM460 cells,the CRC cell lines had significantly higher FMRP expression level.Bioinformatics analysis suggested the involvement of FMRP in regulation of reactive oxygen,oxidative stress-induced cell death,mitochondrial respiration,and glutathione metabolism pathways.In the cell experiments,FMRP knockdown significantly inhibited proliferation of HCT116 cells,lowered cellular GSH content,increased MDA and ROS levels,Fe2+fluorescence intensity,and mitochondrial membrane potential,and decreased SLC7A11/GPX4 protein expressions and the phosphorylation levels of ERK,MEK,MAPK,and RAS proteins;FMRP overexpression resulted in the opposite changes in the cells.In the tumor-bearing nude mice,HCT116 cells with FMRP knockdown showed attenuated tumorigenic potential with lowered xenograft growth rate and reduced SLC7A11 expression in the xenograft.Conclusion The high expression of FMRP inhibits ferroptosis in CRC cells and promotes progression of CRC by activating the RAS/MAPK signaling pathway.

Objective To investigate the mechanism by which fragile X mental retardation protein(FMRP)regulates ferroptosis evasion in colorectal cancer(CRC)cells.Methods We examined FMRP expression levels in CRC cell lines using RT-qPCR and Western blotting and analyzed the biological functions and signaling pathways involved in FMRP-mediated regulation of CRC progression using the TCGA database.A lentiviral FMRP overexpression vector(Lv-FMRP)and 3 knockdown vectors(siFMRP-1,siFMRP-2,and siFMRP-3)were constructed,and their effects on proliferation of HCT116 cells were examined using CCK8 assay and plate clone formation assay;the changes in cell ferroptosis level was determined using MDA/ROS/GSH/Fe2+kits,mitochondrial membrane potential changes were detected using JC-1 fluorescence staining,and the expressions of proteins associated with ferroptosis and the RAS/MAPK signaling pathway were detected using Western blotting.The subcutaneous tumorigenic potential of the transfected cells was evaluated in nude mice.Results Compared with normal colonic mucosal epithelial NCM460 cells,the CRC cell lines had significantly higher FMRP expression level.Bioinformatics analysis suggested the involvement of FMRP in regulation of reactive oxygen,oxidative stress-induced cell death,mitochondrial respiration,and glutathione metabolism pathways.In the cell experiments,FMRP knockdown significantly inhibited proliferation of HCT116 cells,lowered cellular GSH content,increased MDA and ROS levels,Fe2+fluorescence intensity,and mitochondrial membrane potential,and decreased SLC7A11/GPX4 protein expressions and the phosphorylation levels of ERK,MEK,MAPK,and RAS proteins;FMRP overexpression resulted in the opposite changes in the cells.In the tumor-bearing nude mice,HCT116 cells with FMRP knockdown showed attenuated tumorigenic potential with lowered xenograft growth rate and reduced SLC7A11 expression in the xenograft.Conclusion The high expression of FMRP inhibits ferroptosis in CRC cells and promotes progression of CRC by activating the RAS/MAPK signaling pathway.

Objective:To discuss the association between occupational acid fog exposure and accelerated biological aging of the workers,and to clarify its related risk factors.Methods:A total of 341 male workers exposed to occupational acid fog and 201 male workers without occupational exposure were selected as the study subjects,and they were divided into exposure group and control group,respectively.The general informations of the subjects in two groups were collected through questionnaires and physical examinations.The levels of red blood cell count(RBC),platelet count(PLT),albumin(ALB),urea(Urea),creatinine(CR),triglycerides(TG),total cholesterol(TC),glycated hemoglobin(HBA1c),and high-sensitivity C-reactive protein(Hs-CRP)in serum of the subjects in two groups were detected.The Klemera-Doubal method(KDM)was used to construct the composite aging measure,KDM-biological age(BA)(KDM-BA).The model parameters were trained using samples from the 2009 China Health and Nutrition Survey(CHNS)Database to calculate the BA acceleration of the subjects in two groups;stratified analysis based on the population characteristics was conducted to analyze the BA of the subjects in two groups with different population characteristics;generalized linear model was used to analyze the factors influencing BA acceleration due to acid fog exposure.Results:The model parameters were trained using samples from the 2009 CHNS Database,including 8 133 cases aged 20-79 years,of which 3 788 were male.The levels of Urea,CR,HBA1c,ALB,and TC,as well as systolic blood pressure(SBP),total working years,sleep duration,and body mass index(BMI)of the subjects between two groups had significant differences(P<0.05).Compared with control group,the BA acceleration of the subjects in exposure group was significantly increased(P<0.05).In entire population and exposure group,the BA acceleration in the smokers was significantly higher than that in the non-smokers(P<0.05).In entire population,control group,and exposure group,the BA accelerations of the subjects in different BMI groups were significantly decreased with the increase of BMI(P<0.05).Compared with control group,the BA acceleration of the subjects in exposure group was significantly increased(P<0.05),including those under 40 years old,with total working years of 4-7 years,Han nationality,unmarried,smokers,and sleep duration 6-7 h,and with overweight.Acid fog exposure,smoking,and BMI were associated with the BA acceleration(β=0.72,95%CI:0.24-1.21;β=0.59,95%CI:0.11-1.06;β=-0.29,95%CI:-0.35—-0.22).Conclusion:Occupational acid fog exposure may accelerate the biological aging in the workers,and acid fog is a risk factor to accelerate the biological aging of the body.

Objective:To explore the protective mechanism of Xuebijing injection (referred to as Xuebijing) on sepsis-associated acute respiratory distress syndrome (ARDS).Methods:① Animal experiments: 100 mice were randomly(random number) divided into 4 groups, including sham operation (Sham) group, cecal ligation and puncture (CLP) group, CLP+low-dose Xuebijing (L-XBJ) group, and CLP+high-dose Xuebijing (H-XBJ) group. The survival rate, lung histological changes, lung wet/dry (W/D) ratio, cell count and protein concentration in bronchoalveolar lavage fluids (BALF), inflammatory factors levels in serum, oxidative stress indicators, cell apoptosis, and key proteins of HIF-1α/p38 MAPK/NF-κB signaling pathway were measured. ② Cell experiments: Mouse alveolar macrophages (MH-S) were cultured in vitro and divided into 6 groups, including control (Con) group, lipopolysaccharide (LPS) group, LPS+L-XBJ group, and LPS+H-XBJ group, LPS+H-XBJ+ dimethyloxallyl glycine (DMOG, HIF-1α activator) group, LPS+H-XBJ+ 2-methoxyestradiol (2ME2, HIF-1α inhibitor) group. The effects of Xuebijing on inflammatory factors, oxidative stress, and cell apoptosis and their relationship with HIF-1α/p38 MAPK/NF-κB signaling pathway were detected.Results:Xuebijing increased the survival rate of mice with sepsis-associated ARDS, relieved lung tissue damage [lung injury score: CLP group (8.778±0.588), CLP+L-XBJ group (5.833±0.310), and CLP+H-XBJ group (4.750±0.246)], alleviated lung W/D ratio, and decreased pneumonia cell infiltration and protein exudation (all P<0.05). Additionally, Xuebijing treatment also diminished the expression of inflammatory factors (TNF-α, IL-1β, and IL-6), intracellular reactive oxygen species (ROS) accumulation, malondialdehyde (MDA) formation, superoxide dismutase (SOD) depletion, and cell apoptosis in LPS-induced MH-S cells and CLP-induced sepsis-associated ARDS mice (all P<0.05). Furthermore, mechanistic investigation further clarified the effects of Xuebijing on inflammation, oxidative stress, and cell apoptosis through the HIF-1α/p38 MAPK/NF-κB signaling pathway. Conclusions:Xuebijing can exert anti-inflammatory, anti-oxidative, and anti-apoptotic effects by suppressing the HIF-1α/p38 MAPK/NF-κB signaling pathway, thereby conferring protection against sepsis-associated ARDS.

Objective:To investigate the dosimetric differences between conventional IMRT and electron beam conformal radiotherapy (EBCRT) combined with IMRT for post-mastectomy left-sided breast cancer patients.Methods:A total of 20 post-mastectomy left-sided breast cancer patients who were treated in the Ningbo First Hospital from June 2018 to October 2021 were retrospectively studied. The planning target volume (PTV) included the supra-and infra-clavicular regions(PTV sc)and the ipsilateral chest wall (PTV cw), and the prescribed dose was 50 Gy/25 f. All radiotherapy plans were designed using the Varian Eclipse treatment planning system (TPS). After that, the dose distribution of the target volume and the dose exposure of organs at risk (OARs) were compared and analyzed. Results:All the IMRT plans met the clinical requirements, yet 2/20 of the EBCRT combined with IMRT plans were not clinically accepted. For these two patients, the maximum chest wall thickness was 3.7 cm and 4.4 cm each, and the designed electron beam energy was 12 MeV and 15 MeV, respectively. The dose to the ipsilateral lung of these two patients exceeded the institution-specific dose limit standard. For the remaining 18 patients whose chest wall thickness was 3 cm or less, the designed electron beams were 9 MeV or less. All the EBCRT combined with IMRT plans were clinically accepted. The target dose distribution of the conventional IMRT was better than that of the EBCRT combined with IMRT (uniformity index (HI): PTV sc: t = -10.20, P<0.05; PTV cw: t = -9.24, P<0.05; conformal index (CI): PTV all: t = 10.39, P <0.05). For OARs, the V5 Gy, V20 Gy, and Dmean of the ipsilateral lung of EBCRT combined with IMRT were lower than those of IMRT ( t = 5.98, 6.30, 11.30, P <0.05). Specifically, the V25 Gy and Dmean of heart decreased by 8.3% and 4.79 Gy, respectively ( t = 15.23, 15.76, P<0.05), the Dmean of the left anterior descending coronary artery (LADCA) decreased by 44.03% ( t = 11.69, P <0.05), and the V5 Gy and Dmean of the contralateral breast decreased by 7.9% and 0.8 Gy, respectively ( t = 3.66, 4.93, P<0.05). The dosimetric differences of other OARs were not statistically significant ( P > 0.05). Conclusions:For post-mastectomy left-sided breast cancer patients with a chest wall thickness of less than 3 cm, EBCRT combined IMRT can significantly reduce the exposure dose to the heart, the ipsilateral lung, and the contralateral breast, which is beneficial to reducing the potential risk of long-term complications after radiotherapy and can further improve the long-term overall survival rate of patients. For patients with thick chest wall, IMRT plans are more technologically ideal.

Objective: To introduce the application of free-style perforator flap based on aesthetic units to repair facial defect after tumor resection. Methods: By following the concept of free-style perforator flap and the principle of facial aesthetic unit, the design of a free-style perforator flap allowed over any nearby cutaneous vessel chosen purely on the characteristics of its Doppler signal. Conventional knowledge of anatomical landmarks and possible vascular variations were less relevant. A greater freedom in flap selection was gained to recover defect in different forms such as rotation flap, advanced flap and propeller flap, which were all based on free-style perforators. The flap size ranged from 1.5 cm×1.0 cm to 12.0 cm×6.0 cm with the perforator diameter of 0.3-3.0 mm in pedicle, and some of the pedicles are "perforator clusters" . Results: A total of 72 cases underwent surgery, and 68 cases survived completely with satisfactory appearance. 1 case healed two weeks later through dressing due to undesired healing, which result ed from high tension secondary postoperative blooding. 3 cases healed in a delay due to congestion and gained acupuncture treat. Conclusions The free-style perforator flap, which depended on Doppler-discerned perforator and facial aesthetic unit, represents safe, reliable and versatile for repairing facial defect after extended resection, and it not only offers a greater freedom in flap selection but also provides good aesthetic result.

Objective To study the value of free-style perforator flap in repairing nasal defects after tumor resection.Methods On the basis of the guidance of free-style perforator flap design concept and the foundation of vascular localization by Doppler based on the ultrasonic echo intensity,combined with nasal beauty subunits,we designed free-style perforator flap in different forms such as V-Y advancing flap,rotating flap,and propeller flap,which had with free-style perforator in pedicle all.We transplanted the flaps to repair nasal defects after tumor resection.The area of the flaps was between 2 cm x 1.0 cm and 8.0 cm x 5.0 cm,and the diameter of the perforate vessel in the pedicle of flap was between 0.3 mm-3 mm,and some of the pedicles were composed of perforate vessel tube bundle.In order to reduce recurrence rate,the radiotherapy was performed according to the pathology in 1 month after surgery.Results In 31 cases of this group,29 cases were performed and the postoperative shape was good.1 case appeared postoperative hemorrhage,and the epidermis of flap formed blister because of greater tension,and the patient healed 10 days later after extraction the bubble fluid and changing medicine.The flap of 1 case was silted because the venous was blocked,and the acupuncture was treated,and then the flap got delayed union.23 cases received adjuvant radio therapy after surgery and fellowed up from 6 months to 5 years,showing that local profile and color were satisfacfory without tumor recurrence.Conclusions It is worthy of clinical promotion to designing freetype perforator flap and to repair defect after extensive nasal tumor resection on the basis of beauty subunits in nasal and vascular anatomy,which can not only reduce the recurrence rate by postoperative radiotherapy in time,but also realize better cosmetic requirements.

OBJECTIVE: To explore the relationship of formaldehyde exposure, genome-wide DNA methylation, and prevalence of childhood acute lymphocytic leukemia( cALL).METHODS: A case-control study was conducted.Fifty-nine newly diagnosed cALL patients were selected as case group,and 54 orthopedic patients were included in control group.Enzyme-linked immunosorbent assay was used to measure the level of formaldehyde-human serum albumin( FA-HSA) and immunofluorescence method was used to examine the genome-wide DNA methylation level in whole blood.RESULTS: The level of FA-HAS in the blood of the case group was higher than that in the control group( median: 59.61 vs 35.06 fg/L,P < 0.01).Genomic-wide DNA methylation level in the case group was lower than that in the controls[( 2.86 ± 0.31) vs( 3.00 ± 0.28),P < 0.05].Formaldehyde exposure level was not associated with genomic-wide DNA methylation( Spearman correlation coefficient =-0.18,P > 0.05).High FA-HAS level and hypomethylation of genomic-wide DNA were risk factors for cALL onset( P < 0.05).CONCLUSION: Patients with high level of formaldehyde exposure and hypomethylation of genomic-wide DNA have a high risk of cALL.

Objective To investigate the characteristics of distribution and expression profiles of plasma miRNA in childhood acute lymphocytic leukemia (cALL) patients;the association between cALL incidence risk and plasma miRNA levels;the feasibility of plasma miRNA serving as cALL diagnostic biomarker.Methods A total of 111 pairs of newly diagnosed cALL patients and patients with fractures were collected from Shenzhen Children's Hospital,China,between January 2015 and November 2016.Age and sex of the cases and controls were 1 ∶ 1 matched and LNATM miRNA microarray was performed using 4 pairs of cALL and controls selected from the sample population.The expression level of miRNA was validated by real time quantitative PCR.Conditional logistic regression analysis was applied to evaluate the association between miRNA expression levels and the incidence risk of cALL.The receiver operating characteristic curve (ROC) and reclassification analysis were conducted to assess the feasibility of miRNAs serving as biomarkers for cALL.Results A total of 204 differentially expressed miRNA were screened out and let-7f-5p,miR-5100,miR-25-3p and miR-3654 were selected for validation identified according to the inclusion criteria.The expression levels of let-7f-5p,miR-5100 and miR-25-3p in the cALL patients were significantly lower than those of the controls (P＜0.01).After adjusting for confounding factors,3 miRNAs remained significantly associated with the risk of cALL (OR and 95％CI were 0.84 (0.76-0.92),0.81 (0.73-0.90)and 0.81 (0.74-0.89),respectively.Results from both the ROC analysis and reclassification analysis showed that introduction of one or more miRNA to traditional risk factors improved the area under the curve (P＜0.05) and provided additional values to diagnosis (P＜0.01).Conclusion The expression levels of let-7f-5p,miR-5100 and miR-25-3p were significantly associated with the incidence rate of cALL,and these miRNAs might serve as promising biomarkers for cALL.

Objective To study the mechanism of different forms of ST-segment elevation in acute myocardial infarction(AMI),and to investigate the distribution of its traditional Chinese medicine (TCM)syndrome types.Methods Two hundred and twelve hospitalized AMI patients with ST-segment elevation from March of 2015 to July of 2017 were divided into group A and group B.Group A had 102 cases with the elevation of concavity of STsegment,and group B had 110 cases with the elevation of arch of ST-segment.The correlation of ST-segment elevation in different forms with TCM syndrome types was analyzed.Results (1) Patients of group B were usually male,young and middle-aged,with or without short-term medical history of coronary heart disease,and coronary angiography results indicated the stenosis or occlusion of the vessels without collateral circulation.Patients in group A were usually female,aged,with medical history of coronary heart disease,and coronary angiography results indicated the existence of collateral circulation.(2) Group A was dominated by Qi-deficiency and blood-stasis syndrome,and the frequency of its syndrome types was in decreasing sequence:Qi-deficiency and blood-stasis syndrome,Qi-yin deficiency syndrome,heat-toxicity and blood-stasis syndrome,phlegm blended with bloodstasis syndrome,syndrome of cold stagnation in heart vessels.Group B was dominated by heat-toxicity and bloodstasis syndrome,and the frequency of its syndrome types was in decreasing sequence:heat-toxicity and bloodstasis syndrome,Qi-deficiency and blood-stasis syndrome,Qi-yin deficiency syndrome,syndrome of cold stagnation in heart vessels,phlegm blended with blood-stasis syndrome.The difference of the distribution of syndrome types was significant between the two groups (P ＜ 0.01).(3) In respect of the differentiation of deficiency and excess syndromes,group A was dominated by deficiency interweaved with excess syndrome,while group B was dominated by excess syndrome.The difference of the distribution of deficiency and excess syndrome was significant between the two groups (P ＜ 0.01).Conclusion AMI patients with different forms of ST-segment elevation have different TCM syndrome types.The investigation results will provide a new vision for the clinical trial of AMI treated with Chinese medicine integrated with western medicine,and will supply evidence for the syndrome differentiation and treatment of AMI patients with different forms of ST-segment elevation,which will contribute to enhancing clinical efficacy,saving life and improving prognosis.