ObjectiveTo construct a stable monoclonal human embryonic kidney 293 （HEK293） cell line expressing recombinant human coagulation factor Ⅶ （rhFⅦ） and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells， and monoclonal stable transfected cell lines were selected using geneticin （G418）. The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction （RT-PCR）. The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay （ELISA）， and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence， indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8， and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was （1.27±0.09） mg/L， and the highest procoagulant activity was （380.29±13.80）%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.

Objective To investigate the prevalence and influencing factors of work-related musculoskeletal disorders (WMSDs) in neck, shoulders and lower back among power grid workers. Methods A total of 371 workers from a power grid enterprise in Guangdong Province were selected as the research subjects using the convenience sampling method. A cross-sectional survey of WMSDs in the neck, shoulders and waist of the workers was conducted using the modified Musculoskeletal Disorders Questionnaire. Results The overall prevalence of WMSDs among the power grid workers was 62.8%, with prevalence of 37.2%, 33.7% and 32.1% for WMSDs in neck, shoulders and lower back, respectively. Binary logistic regression analysis showed that power grid workers who did not maintain a good sitting posture or were unable to maintain good sitting posture over prolonged periods had a higher risk of neck WMSDs than those who maintained a good sitting posture (both P<0.01). A higher frequency of head-down posture during work was associated with an increased risk of neck WMSDs (both P<0.05). Female workers had a higher risk of shoulder WMSDs than male workers (P<0.05). A higher frequency of early awakening with inability to fall back asleep, as well as a higher frequency of head-down posture during work, were both associated with an increased risk of shoulder WMSDs (all P<0.05). Workers with a sedentary habit had a higher risk of shoulder WMSDs than those without (P<0.01). Workers with insufficient rest time had a higher risk of shoulder WMSDs than those with sufficient rest time (P<0.01). Both maintenance workers and managers had a higher risk of lower back WMSDs than workers in other occupational categories (all P<0.05). Workers who experienced nocturnal awakening with difficulty falling back asleep at frequency of several times per year, ≥1 time /month, ≥1 time /week, had higher risk of lower back WMSDs than those who never experienced such problem (all P<0.05). Workers with a sedentary habit and those uncertain about having a sedentary habit both had a higher risk of lower back WMSDs than those without (both P<0.05). Greater forward neck flexion during work was associated with an increased risk of lower back WMSDs (both P<0.05). Workers who occasionally tilted their necks to the side while working had a higher risk of lower back WMSDs than those who did not (P<0.05). Conclusion WMSDs are highly prevalent among power grid workers, particularly affecting the neck, shoulder, and waist. Influencing factors include individual characteristics, behavioral habits, work organization, work posture, and workload. Maintaining proper posture and adequate rest may reduce the risk of WMSDs.

Objective To explore the effect of platelet-rich fibrin(PRF)combined with sulfadiazine silver dressing on healing time and curative effect of chronic refractory wounds.Methods A total of 126 patients with chronic refractory wounds who were treated in Ningde City Hospital from December 1,2022 to December 1,2023 were selected as research objects.They were randomly divided into control group and experimental group according to odd-even number,with 63 cases in each group.The control group was treated with sulfadiazine silver dressing,and the experimental group was treated with PRF and sulfadiazine silver dressing.The curative effect,wound recovery,serum inflammatory cytokine levels and complications were compared between the two groups.Results The total effective rate of the experimental group was significantly higher than that of the control group([96.83%,61/63]vs[77.78%,49/63],χ2=10.309,P<0.05).The frequency of dressing changes and wound healing time in the experimental group were significantly less than those in the control group([4.44±1.53]times vs[9.16±2.23]times,[15.35±2.29]d vs[20.25±3.24]d,P<0.05).The reduction of wound area in the experimental group was significantly higher than that in the control group([14.95±2.80]cm2 vs[11.21±2.35]cm2,P<0.05).There was no significant difference in serum inflammatory cytokine levels between the two groups before treatment(t=0.209,0.295,0.088,P>0.05).After 14 of treatment,the levels of CRP,PCT,and IL-6 were decreased in both groups,and the levels of CRP,PCT,and IL-6 in the experimental group were significantly lower than those in the control group([1.14±0.86]mg/L vs[2.11±0.75]mg/L,[4.07±0.56]μg/L vs[4.96±0.71]μg/L,[61.54±25.33]ng/L vs[84.48±24.76]ng/L,t=6.748,7.809,5.140,P<0.05).The total incidence of complications in the experimental group was significantly lower than that in the control group(9.52%vs 25.40%,χ2=5.507,P<0.05).Conclusion PRF combined with sulfadiazine silver dressing can increase curative effects,accelerate wound healing,improve serum inflammatory markers,and reduce complications in patients with chronic refractory wounds.

Objective:To analyze by bioinformatics the association between the expression of membrane-associated tyrosine/threonine protein kinase 1(PKMYT1)in glioma with its prognostic value,biological function,drug sensitivity,gene mutation,and immune infiltration.Methods:The differential expression of PKMYT1 was analyzed based on the Chinese Glioma Genome Atlas database(CGGA)and the Cancer Genome Atlas database(TCGA).Pathways likely to be enriched for PKMYT1 were predicted by gene ontology analysis(GO)and gene set enrichment analysis(GSEA).PKMYT1 was subjected to Pearson correlation and gene set variation analysis(GSVA)with cell cycle-related genes and gene sets.The survival prognosis,gene mutation,drug sensitivity and immune infiltration were further analyzed for PKMYT1 high and low expression groups of glioma patients.Results:PKMYT1 was significantly highly expressed in WHO high-grade glioma(P<0.000 1),IDH wild-type glioma(P<0.05),and glioblastoma(P<0.000 1).Overall survival(OS)of patients in the PKMYT1 low expression group was significantly higher than that of the high expression group(P<0.05).Cox regression analysis showed that PKMYT1 expression level was an independent prognostic factor for OS(P<0.05).GO and GSEA analyses showed that the gene sets co-expressed with PKMYT1 were mainly enriched in signaling pathways such as cell cycle,DNA replication and DNA damage repair.Pearson correlation and GSVA analyses showed that the expression of PKMYT1 was significantly and positively correlated with the cell cycle-related genes,gene sets and cell cycle checkpoint genes(P<0.01).Drug sensitivity analysis revealed that patients in the PKMYT1 high expression group had high sensitivity osimertinib,dabrafenib,carmustine and cediranib(P<0.05).Mutation analysis revealed that the IDH1 gene had a higher mutation frequency in the PKMYT1 low expression group.The results of immune infiltration analysis showed that PKMYT1 expression was significantly positively correlated with glioma stroma score(r=0.13,P<0.001),immune score(r=0.11,P<0.01)and ESTIMATE score(r=0.13,P<0.001);and was significantly positively correlated with immune cell infiltration level of regulatory T(Treg)cells and M2-type macrophages(P<0.05).Conclusion:Patients with high PKMYT1 expression have a poorer prognosis,and the mechanism may be related to tumor immune infiltration and cell cycle regulation.PKMYT1 is expected to be a potential target for the diagnosis and treatment of glioma.

Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

OBJECTIVE To analyze the influencing factors of voriconazole trough concentration and adverse drug reactions （ADR） in renal transplant recipients. METHODS Data from inpatients who received voriconazole and therapeutic drug monitoring in our hospital between January 2022 and August 2023 were retrospectively analyzed. Patients were divided into renal transplant group and non-renal transplant group based on transplantation status. A 1∶1 propensity score matching （PSM） method was used to balance differences in baselines between the two groups. Voriconazole trough concentrations， target attainment rate， clinical efficacy， and ADR were compared between the two groups. Multiple linear regression （backward） was used to analyze the factors influencing voriconazole trough concentrations in the renal transplant group. Univariate analysis and binary Logistic regression were used to identify independent risk factors for ADR in the renal transplant group. RESULTS After PSM， 48 patients were included in each group. There were no statistically significant differences in the mean voriconazole trough concentration， target attainment rate or efficacy rate between the two groups （P＞0.05）. The total incidence of ADR was significantly higher in the renal transplant group than in the non-renal transplant group （P＜0.05）. Multiple linear regression analysis showed that age， average daily dose， pulmonary infection， total bilirubin during medication， day-1 loading dose， use of the original drug， concomitant immunosuppressant use， and the occurrence of ADR were factors influencing voriconazole trough concentration in renal transplant patients （P＜0.05）. Binary Logistic regression analysis showed that abnormal direct bilirubin during medication [OR＝7.747， 95%CI （1.334， 45.005）， P＝0.023] was an independent risk factor for ADR in renal transplant patients receiving voriconazole. CONCLUSIONS Age， average daily dose， pulmonary infection， use of the original drug， day-1 loading dose， total bilirubin during medication， concomitant immunosuppressant use， and the occurrence of ADR are the factors influencing voriconazole trough concentration in renal transplant patients. Furthermore， patients with abnormal direct bilirubin during medication are more susceptible to ADR.

Objective To investigate the situation and influencing factors of preserved ratio impaired spirometry (PRISm) among dust-exposed workers in a wooden furniture manufacturing enterprise. Methods A total of 562 dust-exposed workers from a wooden furniture enterprise in Guangdong Province were selected as the study subjects using the convenience sampling method. The result of health-status questionnaire and occupational medical examinations among the participants were studied, and the influencing factors of PRISm were studied using the binary logistic regression analysis method. Results The detection rate of PRISm was 22.1% (124/562) among the study subjects. Binary logistic regression result showed that male workers had a higher risk of PRISm than female workers (P<0.01). Current smokers had a higher risk of PRISm than non-smokers (P<0.05). Workers with longer duration of exposure to occupational hazards had a higher risk of PRISm (P<0.05), those with higher grade of small-airway dysfunction had a higher risk of PRISm (P<0.01). In terms of body mass index, overweight or obese workers showed a higher detection rate of PRISm than those with normal weight (P<0.01). Greater amount of smoking pack per year had a higher risk of PRISm (P<0.01). Conclusion Dust-exposed workers in the wooden furniture manufacturing industry show a relatively high detection rate of PRISm. Male, current smoker, longer duration of exposure to occupational hazards, small airway dysfunction, overweight or obese, and smoking pack per year are influencing factors of PRISm among the dust-exposed workers in the wooden furniture manufacturing industry.

BACKGROUND: The benefits of ideal cardiovascular-health metrics (ICVHMs) in patients with renal insufficiency remain unclear. This study aimed to investigate the associations between ICVHM and prognosis in a renal insufficiency population. METHODS: The trial enrolled 29,682 participants from the US National Health and Nutrition Examination Survey (NHANES), 2007-2018, with mortality follow-up through December 31, 2019. Participants were divided into three groups based on estimated glomerular filtration rates. Cardiovascular health was assessed using new "Life's Essential 8" metrics. Cox regression analyses based on NHANES data were used to determine the associations between ICVHMs and cardiovascular mortality in patients with renal insufficiency. RESULTS: During a mean follow-up of 6.58 years, ideal cardiovascular health (hazard ratio [HR] = 0.42; 95% confidence interval [CI]; 0.25-0.70) and ideal health behavior (HR = 0.53; 95% CI; 0.39-0.73) reduced cardiovascular mortality in participants with renal insufficiency. For each one ICVHM increment, a 25% reduction in cardiovascular mortality was recorded (95% CI; 0.69-0.82). When compared with participants with normal renal function, for those with mild renal insufficiency, the HR for cardiovascular mortality gradually decreased from 1.47 (95% CI; 0.85-2.52) in those who had ≤1 ICVHMs to 0.30 (95% CI; 0.12-0.77) in participants who had >6 ICVHMs. CONCLUSIONS From an ICVHM perspective, enhanced cardiovascular benefits were observed in individuals with renal insufficiency, coupled with a reduced risk of all-cause mortality. Furthermore, when compared with individuals with normal renal function, increased ICVHMs can mitigate adverse risks associated with renal impairment.
Humans ; Male ; Female ; Nutrition Surveys ; Middle Aged ; Renal Insufficiency/physiopathology* ; Aged ; Prognosis ; Adult ; Cardiovascular Diseases/mortality* ; Glomerular Filtration Rate/physiology* ; Proportional Hazards Models

BACKGROUND: Tissue engineering holds promise for vascular repair and regeneration by mimicking the extracellular matrix of blood vessels. However, achieving a functional and thick vascular wall with aligned fiber architecture by electrospinning remains a significant challenge. METHODS: A novel electrospinning setup was developed that utilizes an auxiliary electrode and a spring. The impact of process parameters on fiber size and morphology was investigated. The structure and functions of the scaffolds were evaluated through material characterization and assessments of cellular biocompatibility. RESULTS: The new setup enabled controlled deposition of fibers in different designed orientations. The fabricated small-diameter vascular scaffolds consisted of an inner layer of longitudinally oriented fibers and an outer layer of circumferentially oriented fibers (L + C vascular scaffold). Key parameters, including rotational speed, the utilization of the auxiliary electrode, and top-to-collector distance (TCD) significantly influenced fiber orientation. Additionally, voltage, TCD, feed rate, needle size, auxiliary electrode and collector-auxiliary electrode distance affected fiber diameter and distribution. Mechanical advantages and improved surface wettability of L + C vascular scaffold were confirmed through tensile testing and water contact angle. Cellular experiments indicated that L + C vascular scaffold facilitated cell adhesion and proliferation, with human umbilical vein endothelial cells and smooth muscle cells attaching and elongating along the fiber direction of the inner and outer layer, respectively. CONCLUSION This study demonstrated the feasibility of fabricating fiber-aligned, thick-walled vascular scaffolds using a modified electrospinning setup. The findings provided insights into how the auxiliary electrode, specific collector influenced fiber deposition, potentially advancing biomimetic vascular scaffold engineering.

BACKGROUND: Tissue engineering holds promise for vascular repair and regeneration by mimicking the extracellular matrix of blood vessels. However, achieving a functional and thick vascular wall with aligned fiber architecture by electrospinning remains a significant challenge. METHODS: A novel electrospinning setup was developed that utilizes an auxiliary electrode and a spring. The impact of process parameters on fiber size and morphology was investigated. The structure and functions of the scaffolds were evaluated through material characterization and assessments of cellular biocompatibility. RESULTS: The new setup enabled controlled deposition of fibers in different designed orientations. The fabricated small-diameter vascular scaffolds consisted of an inner layer of longitudinally oriented fibers and an outer layer of circumferentially oriented fibers (L + C vascular scaffold). Key parameters, including rotational speed, the utilization of the auxiliary electrode, and top-to-collector distance (TCD) significantly influenced fiber orientation. Additionally, voltage, TCD, feed rate, needle size, auxiliary electrode and collector-auxiliary electrode distance affected fiber diameter and distribution. Mechanical advantages and improved surface wettability of L + C vascular scaffold were confirmed through tensile testing and water contact angle. Cellular experiments indicated that L + C vascular scaffold facilitated cell adhesion and proliferation, with human umbilical vein endothelial cells and smooth muscle cells attaching and elongating along the fiber direction of the inner and outer layer, respectively. CONCLUSION This study demonstrated the feasibility of fabricating fiber-aligned, thick-walled vascular scaffolds using a modified electrospinning setup. The findings provided insights into how the auxiliary electrode, specific collector influenced fiber deposition, potentially advancing biomimetic vascular scaffold engineering.