Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

Osteoporosis(OP) is a senile bone disease characterized by an imbalance between bone remodeling and bone formation. Targeting pathogenesis of kidney deficiency, spleen deficiency, and blood stasis, Bushen Jianpi Huoxue Decoction has a significant effect on the treatment of OP by tonifying kidney, invigorating spleen, and activating blood circulation. MicroRNA(miRNA) and the anti-apoptotic protein B-cell lymphoma-2-like protein 1(BCL2L1) are closely related to bone cell metabolism. Therefore, in this study, the binding of miR-140-5p to BCL2L1 was detected by dual luciferase assay and polymerase chain reaction(PCR). After silencing or overexpressing miR-140-5p, the apoptosis, autophagy, and osteogenic function of human fetal osteoblast cell line 1.19(HFOB1.19) were observed by flow cytometry and Western blot. Bushen Jianpi Huoxue Decoction-containing serum was prepared by intragastric administration of Bushen Jianpi Huoxue Decoction in rats. Different concentrations of Bushen Jianpi Huoxue Decoction-containing serum were used to treat HFOB1.19 with or without miR-140-5p mimic. The expression of osteogenic proteins in each group was observed, and the role of miR-140-5p/BCL2L1 in apoptosis and autophagy of HFOB1.19 was studied, along with the effect of Bushen Jianpi Huoxue Decoction on these processes. As indicated by the dual luciferase assay, miR-140-5p bound to BCL2L1. Flow cytometry and Western blot showed that miR-140-5p promoted apoptosis and inhibited autophagy in HFOB1.19. After intervention with high, medium, and low doses of Bushen Jianpi Huoxue Decoction-medicated serum, compared with the miR-140-5p NC group, the expression of osteocalcin(OCN), osteopontin(OPN), Runt-related transcription factor 2(RUNX2), and transforming growth factor beta 1(TGF-β1) decreased in the miR-140-5p mimic group, while the expression of bone morphogenetic protein 2(BMP2) showed no significant difference under high-dose intervention. Therefore, miR-140-5p/BCL2L1 can promote apoptosis and inhibit autophagy in HFOB1.19. Bushen Jianpi Huoxue Decoction can affect the osteogenic effect of miR-140-5p through BMP2.
MicroRNAs/metabolism* ; Autophagy/drug effects* ; Apoptosis/drug effects* ; Humans ; Drugs, Chinese Herbal/administration & dosage* ; Animals ; Cell Line ; bcl-X Protein/metabolism* ; Osteoblasts/metabolism* ; Rats ; Osteoporosis/physiopathology* ; Male ; Rats, Sprague-Dawley ; Osteogenesis/drug effects*

Syringa pinnatifolia, belonging to the family Oleaceae, is a species endemic to China. It is predominantly distributed in the Helan Mountains region of Inner Mongolia and Ningxia of China. The peeled roots, stems, and thick branches have been used as a distinctive Mongolian medicinal material known as "Shan-chen-xiang", which has effects such as suppressing "khii", clearing heat, and relieving pain and is employed for the treatment of cardiovascular and pulmonary diseases and joint pain. Over the past five years, significant increase was achieved in research on chemical constituents and pharmacological effects. There were a total of 130 new constituents reported, covering sesquiterpenoids, lignans, and alkaloids. Its effects of anti-myocardial ischemia, anti-cerebral ischemia/reperfusion, sedation, and analgesia were revealed, and the mechanisms of agarwood formation were also investigated. To better understand its medical value and potential of clinical application, this review updates the research progress in recent five years focusing on the chemical constituents and pharmacological effects of S. pinnatifolia, providing reference for subsequent research on active ingredient and support for its innovative application in modern medicine system.
Medicine, Mongolian Traditional ; Humans ; Drugs, Chinese Herbal/pharmacology* ; Animals ; Syringa/chemistry*

In the process of maintaining the steady state of bone tissue, the transcription network and signal pathway of the body play a vital role. These complex regulatory mechanisms need precise coordination to ensure the balance between bone formation and bone absorption. Once this balance is broken, it may lead to pathological changes of bone and cartilage, and then lead to various bone diseases. Therefore, it is of great significance to understand these regulatory mechanisms for the prevention and treatment of bone diseases. In recent years, with the deepening of research, more and more lncRNA has been found to be closely related to bone health. Among them, nuclear paraspeckle assembly transcript 1 (NEAT1), as an extremely abundant RNA molecule in mammalian nuclei, has attracted extensive attention. NEAT1 is mainly transcribed from a specific site in human chromosome 11 by RNA polymerase II (RNaseP), which can form two different subtypes NEAT1_1 and NEAT1_2. These two subtypes are different in intracellular distribution and function, but they participate in many biological processes together. Studies have shown that NEAT1 plays a specific role in the process of cell growth and stress response. For example, it can regulate the development of osteoblasts (OB), osteoclasts (OC) and chondrocytes by balancing the differentiation of bone marrow mesenchymal stem cells (BMSCs), thus maintaining the steady state of bone metabolism. This discovery reveals the important role of NEAT1 in bone development and remodeling. In addition, NEAT1 is closely related to a variety of bone diseases. In patients with bone diseases such as osteoporosis (OP), osteoarthritis (OA) and osteosarcoma (OS), the expression level of NEAT1 is different. These differential expressions may be closely related to the pathogenesis and progression of bone diseases. By regulating the level of NEAT1, it can affect a variety of signal transduction pathways, and then affect the development of bone diseases. For example, some studies show that by regulating the expression level of NEAT1, the activity of osteoclasts can be inhibited, and the proliferation and differentiation of osteoblasts can be promoted, thus improving the symptoms of osteoporosis. It is worth noting that NEAT1 can also be used as a key sensor for the prevention and treatment of bone diseases. When exercising or receiving some natural products, the expression level of NEAT1 will change, thus reflecting the response of bones to external stimuli. This feature makes NEAT1 an important target for studying the prevention and treatment strategies of bone diseases. However, although the role of NEAT1 in bone biology and bone diseases has been initially recognized, its specific mechanism and regulatory relationship are still controversial. For example, the expression level, mode of action and interaction with other molecules of NEAT1 in different bone diseases still need further in-depth study. This paper reviews the role of NEAT1 in maintaining bone and cartilage metabolism, and discusses its expression and function in various bone diseases. By combing the existing research results and controversial points, this paper aims to provide new perspectives and ideas for the prevention and treatment of bone diseases, and provide useful reference and enlightenment for future research.

The polymerase 1 and transcript release factor (PTRF)-cytoplasmic phospholipase A2 (cPLA2) phospholipid remodeling pathway facilitates tumor proliferation in glioma. Nevertheless, blockade of this pathway leads to the excessive activation of oncogenic receptors on the plasma membrane and subsequent drug resistance. Here, CD26/dipeptidyl peptidase 4 (DPP4) was identified through screening of CRISPR/Cas9 libraries. Suppressing PTRF-cPLA2 signaling resulted in the activation of the epidermal growth factor receptor (EGFR) pathway through phosphatidylcholine and lysophosphatidylcholine remodeling, which ultimately increased DPP4 transcription. In turn, DPP4 interacted with EGFR and prevented its ubiquitination. Linagliptin, a DPP4 inhibitor, facilitated the degradation of EGFR by blocking its interaction with DPP4. When combined with the cPLA2 inhibitor AACOCF3, it exhibited synergistic effects and led to a decrease in energy metabolism in glioblastoma cells. Subsequent in vivo investigations provided further evidence of a synergistic impact of linagliptin by augmenting the sensitivity of AACOCF3 and strengthening the efficacy of temozolomide. DPP4 serves as a novel target and establishes a constructive feedback loop with EGFR. Linagliptin is a potent inhibitor that promotes EGFR degradation by blocking the DPP4-EGFR interaction. This study presents innovative approaches for treating glioma by combining linagliptin with AACOCF3 and temozolomide.

Based on the concept of"two-stage,four-phase,and three-phase treatments"of in vitro fertilization and embryo transfer(IVF-ET)assisted the whole-course management of TCM by national TCM master You Zhaoling,this article analyzed the treatment strategy assisted by acupuncture in five aspects:principle,method,prescription,acupoint,and technique.Principle:syndrome differentiation and treatment follow different cases;method establish guiding methods based on principles,while following stage diagnoses and treatments;prescription:determine the prescriptions based on therapeutic principles within the context of overall regulation;acupoint:precise acupoint taking and rational acupoint matching;technique:the primary method is acupuncture,supported by various therapeutic modalities,providing a comprehensive framework and evidence-based support for the clinical application of acupuncture and moxibustion in IVF-ET.

Objective:To analyze the changes in the prevalence characteristics of multimorbidity among adolescents aged 13-18 in Inner Mongolia Autonomous Region from 2019 to 2022 and to explore the association between multimorbidity and moderate to high-intensity physical activity among them.Methods:A stratified random cluster sampling method was used to select students aged 13-18 in Inner Mongolia Autonomous Region every September from 2019 to 2022. Physical examinations, demographic characteristics, and depression-related surveys were conducted to analyze the multimorbidity of overweight, obesity, high blood pressure, myopia, spinal curvature abnormality, and depression. A logistic regression model was used to analyze the association between multimorbidity and moderate to high-intensity physical activity.Results:From 2019 to 2022, 70 972, 62 923, 80 254, and 78 288 study subjects were included, with the rates of multimorbidity being 56.4%, 55.4%, 57.2%, and 55.8%, respectively. The rates of multimorbidity remained relatively stable from 2019 to 2022 ( χ2=0.06, P=0.950). The incidence of multimorbidity among girls was significantly higher than that among boys ( P<0.001). The incidence of multimorbidity among urban students was significantly higher than that among suburban students ( P<0.001). The incidence of multimorbidity among high school students was higher than that among middle school students ( P<0.001). The top three multimorbidity combinations were myopia and overweight/obesity (26.4%), myopia and high blood pressure (24.4%), and myopia and depression (19.8%), while the least common combination was depression and spinal curvature abnormality (1.1%). The multimorbidity patterns showed no significant differences between years ( χ2=0.03, P=0.999). The multimorbidity status was significantly associated with the status of meeting the standard of moderate to high-intensity physical activity ( OR=0.83, 95% CI: 0.80-0.86). The association was stronger in boys ( OR=0.77, 95% CI: 0.73-0.81) compared with girls ( OR=0.90, 95% CI: 0.85-0.96), with a significant interaction term ( P<0.001). Conclusion:From 2019 to 2022, the incidence of multimorbidity among adolescents aged 13 to 18 in Inner Mongolia Autonomous Region is relatively high, mainly due to the co-occurrence of myopia and other health problems. Adequate physical activity is an important factor in reducing multimorbidity.

Objective:To explore the characteristics and evolutionary features of cognitive impairment and clinical transformation in early-stage Parkinson′s disease (PD) patients.Methods:Based on the cohort of patients with primary unmedicated PD admitted to the Parkinson′s Specialized Outpatient Clinic of Affiliated Brain Hospital of Nanjing Medical University from November 2018 to July 2022, follow-up was conducted for PD patients who completed the baseline assessment and had a follow-up time of 1.5 years or more, and a total of 87 patients finally completed the follow-up and were included in the study. At follow-up, the 87 patients were divided into a cognitively impaired group ( n=36) and a cognitively normal group ( n=51) according to the norm proposed by Professor Jia Jianping and colleagues in 2011 for the Chinese elderly population. Differences in baseline clinical characteristics between the 2 groups were compared, and binary Logistic regression analysis was used to explore risk factors for cognitive impairment in PD patients. In addition, transformed grouping according to cognitive assessment results at baseline and follow-up was used to compare differences in patients′ baseline clinical characteristics among the 3 groups: a reversal group [Parkinson′s disease-mild cognitive impairment (PD-MCI), reverting to Parkinson′s disease-cognitively normal (PD-CN); n=15], a non-reversal group (persistent PD-MCI; n=24), and a stable group (stable PD-CN; n=36). Results:Cognitive reversal occurred at follow-up in 36.6% (15/41) of patients with cognitive impairment at baseline, and 21.7% (10/46) of patients with normal cognition at baseline had cognitive impairment at follow-up. At the end of the follow-up, the 87 patients with PD had higher Unified Parkinson′s Disease Rating Scale Ⅱ (UPDRS-II) scores [8 (6, 11)], Unified Parkinson′s Disease Rating Scale Ⅲ (UPDRS-Ⅲ) scores [23 (16, 30)], and Hoehn-Yahr stages [2.0 (1.5, 2.5)] than those at baseline [7(4, 10), 19(14, 25), 1.5(1.0, 2.0)]. The differences were statistically significant ( Z=-2.498, P=0.012; Z=-3.039, P=0.002; Z=-4.436, P<0.001). The cognitively impaired group had lower Montreal Cognitive Assessment scores [22.00(19.00, 23.75)] and fewer years of education [9.00(8.00, 11.75) years] but higher Parkinson′s Disease Non-Motor Symptoms Questionnaire (PD-NMSQ) scores [8.00(5.25, 12.00)] than the cognitively normal group [25.00(24.00, 27.00), 12.00(9.00, 15.00) years, 6.00(3.00, 8.00)], and the differences were statistically significant ( Z=-4.764, P<0.001; Z=-3.016, P=0.003; Z=-3.281, P=0.001). Multivariate Logistic regression showed that years of education ( OR=0.829, 95%CI 0.715-0.960, P=0.012) and PD-NMSQ scores ( OR=1.200, 95%CI 1.040-1.384, P=0.012) were independent predictors of cognitive impairment in patients with PD. There were statistically significant differences among the reversal, non-reversal, and stable groups in years of education ( F=5.366, P=0.010), PD-NMSQ scores ( H=10.795, P=0.005), and UPDRS-Ⅱ scores ( H=6.957, P=0.031). Pairwise comparisons showed lower PD-NMSQ scores [4.00(3.00, 7.00) vs 8.00(6.25, 12.75); Z=-2.989, P=0.003] and lower UPDRS-Ⅱ scores [6.00(3.00, 6.00) vs 7.00(6.00, 10.00); Z=-2.756, P=0.006] in the reversal group than in the non-reversal group, indicating better baseline quality of life in cognitive reversal patients. Conclusions:Low educational level and severe non-motor symptoms were risk factors predicting cognitive impairment in PD patients. Conversely, mild non-motor symptoms with high quality of life (lower UPDRS-Ⅱ scores) were important factors for cognitive reversal.

Aim To explore the effect of the classical famous prescription Dahuang Zhechong pill(DHZCP)on relieving liver cirrhosis by regulating the stress fiber remodeling mediated by mechanistic signaling pathway and to explore the underlying mechanism.Methods Mice were randomly divided into the control group,model group,DHZCP low-dose group,DHZCP high-dose group,and Colchicine-positive control group.The liver cirrhosis mouse model was constructed by intrap-eritoneal injection of olive oil-solubilized CCl4.HE staining and serologic markers were used to reflect liver injury.Masson staining was used to evaluate collagen deposition in liver tissue.ELISA was applied to detect vasoactive molecules and cancer indicators.Atomic force microscopy was employed to detect liver tissue stiffness.Color Doppler diagnostic instrument was used to assess portal blood flow velocity.Western blot was utilized to detect ROCK2 expression and phosphoryla-tion of YAP,Cofilin,and MLC.Results The liver tis-sues in the model group had obvious inflammatory cell infiltration and collagen deposition,accompanied by significant elevation of serum transaminases and fibrosis indexes.Similarly,vasoactive molecules and cancer in-dicators were elevated,and the mechanoregulatory pro-tein ROCK2 expression and phosphorylation of Cofilin and MLC were elevated,with YAP being strongly de-phosphorylated.Both low and high doses of DHZCP re-versed the pathological changes,serological indices,and inhibited the activation of the stress fiber(SF)re-modeling mechanistic signaling pathway.Conclusion DHZCP effectively ameliorates liver tissue lesions in mice with liver cirrhosis,and its mechanism may be re-lated to the inhibition of SF remodeling mechanistic signaling pathway.

Objective:To investigate exercise fear condition and its influencing factors in patients with coronary heart dis-ease(CHD)involving in phase Ⅱ cardiac rehabilitation after percutaneous coronary intervention(PCI).Methods:Conven-ience sampling was performed among cardiac rehabilitation patients from a Shanxi grade-A tertiary hospital between Janu-ary 2023 and June 2023.General data questionnaire,Tampa Scale for Kinesiophobia-Short Version for Heart patients(TSK-SV H),the Multidimensional Scale of Perceived Social Support(MSPSS)and the Post Percutaneous Coronary In-tervention Health Literacy Scale(PPCIHLS)were used for assessment.Pearson and Spearman rank correlation analyses were applied to investigate the relationships between social support factors and health literacy factors with patients'fear of exercise;multivariate linear regression models and quantile regression models were used to analyze the influencing factors of fear of exercise.Results:Among the 118 patients,average TSK-SV H score was(33.78±3.79)points;10.2％patients showed significant fear.The correlation analysis showed that PPCIHLS score was negatively correlated with TSK-SV H score(r=-0.423,P＜0.001).Results from multivariate regression analysis indicated that health literacy,cardiac func-tion class and drinking status affected the average score of exercise fear(P＜0.05 all).The quantile regression model pro-vided additional insights,showing that the impact of factors such as living situation,drinking status,cardiac function class and health literacy on exercise fear varied across different quantiles.Notably,cardiac function class had a consistently posi-tive effect on TSK-SV H score at various quantiles.Conclusion:Improving health literacy and tailored rehabilitation plans are vital to reduce exercise fear and improve cardiac rehabilitation effect.