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 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 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 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 efficacy and safety of 450 nm blue laser with 6 o’clock positioning in the treatment of middle lobe hyperplasia of prostate, in order to promote the clinical application of this surgery. 【Methods】 Clinical data of 20 patients with middle lobe hyperplasia of prostate treated with 450 nm blue laser with 6 o’clock positioning during Mar.and Aug.2023 were retrospectively analyzed.The operation time, postoperative bladder irrigation time, catheter indwelling time, hospital stay, and complications were recorded.The maximum urinary flow rate (Qmax), post-void residual volume (PVR), quality of life scale (QoL), international prostate symptom score (IPSS) before surgery and 1 month after surgery were compared. 【Results】 The operation time was (26.80±7.22) min, and bladder irrigation time was (20.50±1.79) h.The catheter was removed on the next day after surgery and all patients were discharged 2 days after operation.Compared to preoperative, one month after surgery, the Qmax [(7.40±1.05) mL/s vs.(19.60±1.76) mL/s] was significantly higher, PVR [(73.50±12.26) mL vs.(9.25±4.94) mL], QoL [(4.55±1.19) vs.(1.95±0.95)], and IPSS [(26.55±1.88) vs.(10.05±1.36)] were significantly lower, the differences being statistically significant (P<0.05).No complications occurred during operation and 1-month follow-up. 【Conclusion】 The 450 nm blue laser with 6 o’clock positioning is a new, safe and effective surgical treatment of middle lobe hyperplasia of prostate, which is worthy of clinical promotion and application.

Objective To explore the effects of intravenous thrombolysis combined with Solitaire FR stent thrombectomy on vascular recanalization,neurologic function and prognosis in patients with acute cerebral infarction(ACI)due to large artery occlusion(LAO).Methods A total of 172 patients with ACI-LAO treated in our department between October 2020 and March 2023 were retrospectively enrolled.According to treatment regimens,they were assigned into control group(86 cases,alteplase intravenous thrombolysis)and study group(86 cases,alteplase intravenous thrombolysis combined with Solitaire FR stent thrombectomy).Vascular recanalization,neurolog-ic function,cerebral perfusion and occurrence of adverse events were compared between the two groups.After 90 d of follow-up,their prognosis was evaluated with modified Rankin scale.Results There was no significant difference in success rate of vascular recanalization between the two group(P＞0.05),but complete recanalization rate was statistically higher in the study group than the control group(68.60％vs 50.00％,P＜0.05).The study group had obviously lower NHISS scores at 7 and 14 d after treatment,higher cerebral blood volume and cerebral blood flow,but shorter mean transit time when compared with the control group(P＜0.05,P＜0.01).No notable difference was observed in the total incidence of adverse events between them(P＞0.05).After 90 d of follow-up,the proportion of good prognosis was higher in the study group than the control group(80.23％vs 63.95％,P＜0.05).Conclusion Intravenous thrombolysis combined with Soli-taire FR stent thrombectomy shows better efficacy in ACI-LAO patients,with better vascular re-canalization and great improvements in neurologic function and prognosis.

Objective:To observe the clinical features and disease outcomes of patients with fundus lesions associated with novel coronavirus infection (COVID-19).Methods:A case series observational study was conducted.Eighteen eyes of 10 patients with COVID-19 related fundus lesions diagnosed in the First Affiliated Hospital of Zhengzhou University from December 2022 to February 2023 were included.The affected eyes were examined by best corrected visual acuity (BCVA), intraocular pressure, slit lamp microscopy, scanning laser fundus photography (SLO), infrared fundus photography, optical coherence tomography (OCT), visual field and microperimetry.After the diagnosis was confirmed, according to the patients' willingness, 6 patients were treated with mecobalamin, vinblastine, and Aescuven forte orally.The other 4 patients were treated with glucocorticoid and methylprednisolone tablets orally with an initial dose of 32 mg/d, and the dose was reduced by 4 mg every 5 days, and potassium chloride tablets, calcium carbonate tablets, and omeprazole enteric-coated capsules were taken orally.According to the diagnosis and treatment of acute macular neuroretinopathy (AMN), the patients were divided into glucocorticoid treatment group (4 cases, 7 eyes) and non-glucocorticoid treatment group (4 cases, 8 eyes).The patients were followed up for 4 weeks.The BCVA, retinal morphology and structure, retinal sensitivity and fixation stability were compared before and after treatment.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University (No.2022-KY-1482-002).Written informed consent was obtained from each subject.Results:Among the enrolled 10 cases (18 eyes), there were 9 cases (17 eyes) of AMN, including 1 eye with Purtscher-like retinopathy (PLR), 1 eye with acute para-central medial maculopathy (PAMM), and 1 eye with multiple transient white dot syndrome (MEWDS).The SLO images of AMN patients showed redbrown irregular lesions in the fovea or parafovea of the macular area.The OCT images showed patchy strong reflection between the OPL and ONL in the fovea or parafovea of the macular area, and the adjacent EZ/IZ had different degrees of local fracture, and the reflection signal was disordered.The BCVA of the eyes after 4 weeks of treatment was higher than that before treatment, and the difference was statistically significant ( Z=-2.823, P<0.05).After 4 weeks of treatment, the retinal sensitivity of the eyes was (26.57±2.24)dB, which was significantly higher than that before treatment (24.17±2.73)dB ( t=-11.329, P<0.001).There was no statistically significant difference in LogMAR BCVA and retinal sensitivity between the AMN glucocorticoid treatment group and the AMN non-glucocorticoid treatment group before and after 4 weeks of treatment ( Z=-0.986, P>0.05; t=-1.656, P>0.05). Conclusions:The main manifestations of COVID-19 related fundus lesions are AMN, PAMM, PLR, MEWDS, etc.OCT, microperimetry and other auxiliary examinations can help to diagnose the disease.The visual impairment and fundus structural changes caused by COVID-19 gradually improve after 4 weeks of treatment.

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.