Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors, including non-small cell lung cancer (NSCLC). However, its detailed molecular mechanism has not been adequately demonstrated. In this research, it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft (PDX) model. Mechanistically, employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis (MST), microRNA-145-5p (miR-145-5p) was pinpointed as a critical target through which elemene exerts its anti-tumor effects. Interestingly, elemene serves as a binding stabilizer for miR-145-5p, demonstrating a strong binding affinity (dissociation constant (K _D) = 0.39 ± 0.17 μg/mL) and preventing its degradation both in vitro and in vivo, while not interfering with the synthesis of the primary microRNA transcripts (pri-miRNAs) and precursor miRNAs (pre-miRNAs). The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA, subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated protein kinase kinase kinase 3 (MAP3K3)/nuclear factor kappaB (NF-κB) pathway. Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.

Hemoglobin A1c (HbA1c) has a unique structure that makes monoclonal antibody (mAb) preparation challenging. This study aims to develop a method for preparing HbA1c mAbs and establish a fluorescent immunochromatographic assay (FICA) for rapid detection of HbA1c. Three glycosylated peptides were synthesized and used to prepare complete antigens, which were identified by dot enzyme-linked immunosorbent assay (Dot-ELISA) and ultraviolet absorption spectroscopy. The complete antigens and natural HbA1c were used for cross-immunization of mice, and the optimal complete antigen was selected. The mouse with the highest serum titer was chosen for mAb preparation. The purity and specificity of the mAbs were verified, and a FICA method was developed. The optimal complete antigen, with a titer of 1:512 000, was successfully prepared and selected. Fusion with splenocytes resulted in four specific HbA1c antibodies (purity > 90%). The best antibody exhibited a binding constant (Ka) of 1.67×10¹⁰ L/mol with the antigen. Based on this antibody, a FICA method was successfully established, capable of producing results within 15 min. The method demonstrated a good linear range (3%-13% HbA1c, y=0.071 3x+0.005 6, R²=0.993 7), recovery rates of 98%-102%, precision < 10.00%, and no nonspecific reactions. Clinical testing of 210 samples showed positive agreement of 96.36%, negative agreement of 97.00%, and overall agreement of 96.68%. The receiver operating characteristic (ROC) curve analysis yielded an area under curve (AUC) of 0.980 9 [95% confidence interval (CI): 0.961 0-1.000 0], with high consistency verified in multicenter studies. We successfully developed a key technique for preparing HbA1c monoclonal antibodies and established a FICA method for rapid detection of HbA1c. It will provide an efficient and convenient detection method for the early diagnosis and long-term management of diabetes and its complications.
Antibodies, Monoclonal/biosynthesis* ; Animals ; Mice ; Glycated Hemoglobin/immunology* ; Mice, Inbred BALB C ; Humans ; Antibody Specificity ; Chromatography, Affinity/methods* ; Enzyme-Linked Immunosorbent Assay/methods* ; Female

Objective:To analyze the clinical data and genetic characteristics of a child with CLN1 neuronal ceroid lipofuscinosis in conjunct with hereditary hyperferritinemia cataract syndrome (HHCS).Methods:A child who was admitted to the PICU of the First Affiliated Hospital of Zhengzhou University in November 2020 was selected as the study subject. Clinical data of the child was collected. Genetic testing was carried out for the child, and the result was analyzed in the light of literature review to explore the clinical and genetic characteristics to facilitate early identification.Results:The patient, a 3-year-old male, had mainly presented with visual impairment, progressive cognitive and motor regression, and epilepsy. Cranial magnetic resonance imaging revealed deepened sulci in bilateral cerebral hemispheres, and delayed myelination. The activity of palmitoyl protein thioesterase was low (8.4 nmol/g/min, reference range: 132.2 ~ 301.4 nmol/g/min), whilst serum ferritin was increased (2 417.70 ng/mL, reference range: 30 ~ 400 ng/mL). Fundoscopy has revealed retinal pigment degeneration. Whole exome sequencing revealed that he has harbored c. 280A>C and c. 124-124+ 3delG compound heterozygous variants of the PPT1 gene, which were respectively inherited from his father and mother. Neither variant has been reported previously. The child has also harbored a heterozygous c. -160A>G variant of the FTL gene, which was inherited from his father. Based on the clinical phenotype and results of genetic testing, the child was diagnosed as CLN1 and HHCS. Conclusion:The compound heterozygous variants of the PPT1 gene probably underlay the disorders in this child. For children with CLN1 and rapidly progressing visual impairment, ophthalmological examination should be recommended, and detailed family history should be taken For those suspected for HHCS, genetic testing should be performed to confirm the diagnosis.

Objective:To explore the risk factors of insomnia among employees in the thermal power generation industry and the network relationships between their interactions, and to provide scientific basis for personalized interventions for high-risk groups with insomnia.Methods:In November 2022, 860 employees of a typical thermal power generation enterprise were selected as the research subjects by cluster sampling. On-site occupational health field surveys and questionnaire surveys were used to collect basic information, occupational characteristics, anxiety, depression, stress, occupational stress, and insomnia. The interaction between insomnia and occupational health psychological factors was evaluated by using structural equation model analysis and Bayesian network construction.Results:The detection rates of anxiety, depression and stress were 34.0% (292/860), 32.1% (276/860) and 18.0% (155/860), respectively. The total score of occupational stress was (445.3±49.9) points, and 160 workers (18.6%) were suspected of insomnia, and 578 workers (67.2%) had insomnia. Structural equation model analysis showed that occupational stress had a significant effect on the occurrence of insomnia in thermal power generation workers (standardized load coefficient was 0.644), and occupational health psychology had a low effect on insomnia (standardized load coefficient was 0.065). However, the Bayesian network model further analysis found that anxiety and stress were the two parent nodes of insomnia, with direct causal relationships, the arc strength was-8.607 and -15.665, respectively. The model prediction results showed that the probability of insomnia occurring was predicted to be 0 in the cases of no stress and anxiety, low stress without anxiety, and no stress with low anxiety. When high stress with low anxiety and low stress with high anxiety occurred, the predicted probability of insomnia occurring were 0.38 and 0.47, respectively. When both high stress and high anxiety occurred simultaneously, the predicted probability of insomnia occurring was 0.51.Conclusion:Bayesian network risk assessment can intuitively reveal and predict the insomnia risk of thermal power generation workers and the network interaction relationship between the risks. Anxiety and stress are the direct causal risks of insomnia, and stress is the main risk of individual insomnia of thermal power generation workers. The occurrence of insomnia can be reduced based on scientific intervention of stress conditions.

Objective:To investigate the protective effect of quercetin against LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, providing valuable insights into the use of quercetin as a virulence inhibitor for Pseudomonas aeruginosa infection treatment. Methods:This was an experimental study. The experimental strain was the standard strain. The LasB protein was obtained utilizing protein recombination technology, while the enzyme activity of LasB was assessed through both the Elastin Congo red assay and fluorescently labelled elastin assay. The LasB-induced THP-1 macrophage infection model was established, and quercetin was utilized for intervention. Cell viability was evaluated via CCK-8 assay, while cell morphology was observed under an inverted microscope. Apoptosis detection involved employing both TUNEL and Annexin V/PI staining. The mRNA expression and protein levels of inflammatory cytokines and COX-2 were determined by RT-qPCR and ELISA respectively. Intracellular ROS levels were quantified using the DCFH-DA fluorescent probe. One-way ANOVA was used for statistical analysis, and Tukey test was used for multiple comparisons. Results:The pLasB with a molecular weight of 33 000 and acceptable enzymatic activity (purity>90%), was successfully obtained. THP-1 macrophages treated with pLasB at a concentration of 100 μg/ml presented significantly decreased viability and integrity rate when compared with the normal control group. Additionally, pLasB promoted apoptosis, up-regulated the levels of inflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12, and TNF-α, increased intracellular ROS fluorescence intensity, and elevated COX-2 mRNA expression level. Furthermore, the viability of THP-1 macrophages was significantly enhanced under quercetin intervention at concentrations of 2.5 μmol/L, 5 μmol/L and 10 μmol/L. The apoptosis rate exhibited a significant reduction from 18.32%±0.17% to 13.17%±0.20%, 11.43%±0.06% and 7.74%±0.04%, respectively ( F=1 679, P<0.05). There was a notable down-regulation of pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-12 and TNF-α while the anti-inflammatory cytokine IL-10 showed a significant up-regulation. Both intracellular ROS fluorescence intensity ( F=86.92, P<0.05) and COX-2 level ( F=24.62, P<0.05) demonstrated a substantial decrease. Conclusion:Quercetin demonstrates significant efficacy in inhibiting LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, which highlights immense potential as a potent virulence inhibitor of Pseudomonas aeruginosa.

Objective:To explore the risk factors of insomnia among employees in the thermal power generation industry and the network relationships between their interactions, and to provide scientific basis for personalized interventions for high-risk groups with insomnia.Methods:In November 2022, 860 employees of a typical thermal power generation enterprise were selected as the research subjects by cluster sampling. On-site occupational health field surveys and questionnaire surveys were used to collect basic information, occupational characteristics, anxiety, depression, stress, occupational stress, and insomnia. The interaction between insomnia and occupational health psychological factors was evaluated by using structural equation model analysis and Bayesian network construction.Results:The detection rates of anxiety, depression and stress were 34.0% (292/860), 32.1% (276/860) and 18.0% (155/860), respectively. The total score of occupational stress was (445.3±49.9) points, and 160 workers (18.6%) were suspected of insomnia, and 578 workers (67.2%) had insomnia. Structural equation model analysis showed that occupational stress had a significant effect on the occurrence of insomnia in thermal power generation workers (standardized load coefficient was 0.644), and occupational health psychology had a low effect on insomnia (standardized load coefficient was 0.065). However, the Bayesian network model further analysis found that anxiety and stress were the two parent nodes of insomnia, with direct causal relationships, the arc strength was-8.607 and -15.665, respectively. The model prediction results showed that the probability of insomnia occurring was predicted to be 0 in the cases of no stress and anxiety, low stress without anxiety, and no stress with low anxiety. When high stress with low anxiety and low stress with high anxiety occurred, the predicted probability of insomnia occurring were 0.38 and 0.47, respectively. When both high stress and high anxiety occurred simultaneously, the predicted probability of insomnia occurring was 0.51.Conclusion:Bayesian network risk assessment can intuitively reveal and predict the insomnia risk of thermal power generation workers and the network interaction relationship between the risks. Anxiety and stress are the direct causal risks of insomnia, and stress is the main risk of individual insomnia of thermal power generation workers. The occurrence of insomnia can be reduced based on scientific intervention of stress conditions.

Objective:To investigate the protective effect of quercetin against LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, providing valuable insights into the use of quercetin as a virulence inhibitor for Pseudomonas aeruginosa infection treatment. Methods:This was an experimental study. The experimental strain was the standard strain. The LasB protein was obtained utilizing protein recombination technology, while the enzyme activity of LasB was assessed through both the Elastin Congo red assay and fluorescently labelled elastin assay. The LasB-induced THP-1 macrophage infection model was established, and quercetin was utilized for intervention. Cell viability was evaluated via CCK-8 assay, while cell morphology was observed under an inverted microscope. Apoptosis detection involved employing both TUNEL and Annexin V/PI staining. The mRNA expression and protein levels of inflammatory cytokines and COX-2 were determined by RT-qPCR and ELISA respectively. Intracellular ROS levels were quantified using the DCFH-DA fluorescent probe. One-way ANOVA was used for statistical analysis, and Tukey test was used for multiple comparisons. Results:The pLasB with a molecular weight of 33 000 and acceptable enzymatic activity (purity>90%), was successfully obtained. THP-1 macrophages treated with pLasB at a concentration of 100 μg/ml presented significantly decreased viability and integrity rate when compared with the normal control group. Additionally, pLasB promoted apoptosis, up-regulated the levels of inflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12, and TNF-α, increased intracellular ROS fluorescence intensity, and elevated COX-2 mRNA expression level. Furthermore, the viability of THP-1 macrophages was significantly enhanced under quercetin intervention at concentrations of 2.5 μmol/L, 5 μmol/L and 10 μmol/L. The apoptosis rate exhibited a significant reduction from 18.32%±0.17% to 13.17%±0.20%, 11.43%±0.06% and 7.74%±0.04%, respectively ( F=1 679, P<0.05). There was a notable down-regulation of pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-12 and TNF-α while the anti-inflammatory cytokine IL-10 showed a significant up-regulation. Both intracellular ROS fluorescence intensity ( F=86.92, P<0.05) and COX-2 level ( F=24.62, P<0.05) demonstrated a substantial decrease. Conclusion:Quercetin demonstrates significant efficacy in inhibiting LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, which highlights immense potential as a potent virulence inhibitor of Pseudomonas aeruginosa.

Objective To investigate the effect and underlying mechanism of roxadustat on apoptosis and senescence of renal tubular epithelial cell line HK-2 induced by heat stress.Methods After HK-2 cells were treated with different concentrations of roxadustat(10,20,30,40 and 50 μmol/L)for 24 h,CCK-8 assay was used to determine the optimal intervention concentration of roxadustat.HK-2 cells were divided into 4 groups(n=3):control group,roxadustat group(30 μmol/L,24 h),heat-stress group(43 ℃,2 h),and heat-stress+roxadustat group(30 μmol/L roxadustat treatmnet for 24 h followed by heat-stress 2 h).Cell viability was detected by CCK-8 assay.Expression of hypoxia-inducible factor-1α(HIF-1α),Cleaved Caspase-3,p16 and p21 at protein level was detected by Western blotting.Immunofluorescence assay was employed to observe the distribution of HIF-1α.β-galactosidase staining kit was utilized to detect SA-β-Gal activity.TUNEL staining was used to measure cell apoptosis.Results The highest cell viability was observed in the cells after 30 μmol/L roxadustat treatment.Heat stress resulted in a significant decrease in cell viability(P＜0.05),elevated protein levels of HIF-1α,Cleaved Caspase-3,p16 and p21(P＜0.05),enhanced SA-β-Gal activity(P＜0.05)and increased percentage of TUNEL-positive cells(P＜0.05)when compared with the cells in the control group.In comparison with the heat-stress group,the heat-stress+roxadustat group showed significant decrease in the protein levels of Cleaved Caspase-3,p16 and p21(P＜0.05),reduced activity of SA-β-Gal[(65.44±5.00)％vs(77.15±2.61)％,P＜0.05]and decreased percentage of TUNEL-positive cells[(16.73±2.20)％vs(46.40±13.87)％,P＜0.05],but increase in cell viability[(86.33±4.51)％vs(66.33±8.50)％,P＜0.05]as well as HIF-1α protein expression(P＜0.05).Furthermore,immunofluorescence assay showed that HIF-1α was mainly distributed in the nucleus and perinucleus.Conclusion Roxadustat attenuates heat stress-induced apoptosis and senescence of renal tubular epithelial cells by upregulating HIF-1α.

Objective To investigate the effect of perineal nerve electroacupuncture combined with tolterodine in the treatment of female overactive bladder(OAB).Methods A total of 150 female OAB patients treated in Huzhou Hospital of Traditional Chinese Medicine,Zhejiang Chinese Medical University from April 2020 to May 2021 were selected and divided into control group and study group according to random number table method,with 75 cases in each group.The control group was treated with tolterodine,and the study group was treated with pudendal nerve electroacupuncture combined with tolterodine.Clinical symptoms,urinary symptoms,and urodynamic indicators[maximum cystometric capacity(MCC),maximum urethral closure pressure(Pura clos max),maximum detrusor pressure(Pdet max),and maximum urine flow rate(Qmax)],bladder function indicators(initial bladder sensation volume,bladder volume during strong micturition desire),nerve growth factor(NGF)mediated transient receptor potential(TRP)pathway indicators[NGF,NGF/urine creatinine(UCr),TRPV1,TRPV4]and clinical efficacy of two groups were compared.Results After treatment,the number of night urination and urgent urination in study group were significantly less than those in control group,and the average volume of urination was significantly more than that in control group(P＜0.05).The overactive bladder symptom scores and visual analogue scale scores of study group were significantly lower than those of control group(P＜0.05).The MCC and Pura·clos·max in study group were significantly higher than those in control group,while Pdet max and Qmax were significantly lower than those in control group(P＜0.05).The initial bladder sensation volume and bladder volume during strong micturition desire in study group were higher than those in control group(P＜0.05).NGF,NGF/UCr,positive expression rates of TRPV1 and TRPV4 in study group were lower than those in control group(P＜0.05).The total effective rate was significantly higher in study group than in control group,(x2=5.374,P=0.020).There was no significant difference in the incidence of total adverse reactions between two groups(x2=0.362,P=0.547).Conclusion Perineal nerve electroacupuncture combined with tolterodine can significantly improve bladder function and inhibit clinical symptoms in female OAB patients,possibly by inhibiting bladder sensitivity caused by NGF-mediated increase of TRP channel proteins TRPV1 and TRPV4.

Pancreatic cancer is a malignant tumor of digestive system with poor prognosis. It has the characteristics of low early diagnosis rate, low feasible operation rate and low treatment success rate. Most patients are in advanced stage when they are found. Chemotherapy and immunotherapy are important treatments for pancreatic cancer because of its low operative rate. However, the extracellular matrix (ECM) of pancreatic cancer forms a special physical and metabolic barrier, which reduces the effect of chemotherapy and immunotherapy. Cancer associated fibroblast (CAF) are tumor promoting cells in the extracellular matrix and are novel targets for anti-tumor interventions. However, the lack of specific markers and unknown differences between CAF subtypes have hindered relevant clinical trials. This article reviews the origin and functional diversity of CAF, and discusses the clinical progress and treatment strategies of CAF and pancreatic cancer.