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.

ObjectiveTo investigate the effect of Niuhuang Qingxinwan （NHQXW） in improving intracerebral hemorrhage （ICH） with Tanre Fushi （phlegm-heat and fu-organ excess） syndrome by maintaining blood-brain barrier （BBB） integrity， and to explore its potential mechanism. MethodsMale mice were administered with 15% autologous feces for 3 consecutive days to simulate spontaneous Tanre Fushi syndrome， followed by surgical induction of collagenase-induced ICH on the fourth day. Mice were randomly assigned to seven groups： Sham， Sham+NHQXW-H， collagenase， collagenase+feces， and NHQXW intervention groups at low （NHQXW-L， 0.225 g·kg^-1）， medium （NHQXW-M， 0.45 g·kg^-1）， and high （NHQXW-H， 0.9 g·kg^-1） doses. Treatments were administered for 3 days after surgery. NHQXW effects on Tanre Fushi syndrome were assessed via fecal water content and small intestinal carbon propulsion rate. Protective effects of NHQXW against ICH with Tanre Fushi syndrome were evaluated by measuring hematoma volume， neurological deficits， and brain water content. BBB integrity was further assessed using Evans blue staining， hematoxylin-eosin （HE） staining， immunofluorescence， and Western blot for Claudin-5， plasmalemma vesicle-associated protein （PLVAP）， matrix metalloproteinase （MMP）-2， and MMP-9. The potential mechanism of NHQXW was investigated by detecting protein expression of protein kinase B （Akt）， extracellular signal-regulated kinase 1/2 （ERK1/2）， signal transducer and activator of transcription 3 （STAT3）， Yes-associated protein （YAP）， and their phosphorylated forms. ResultsCompared with the collagenase+feces group， NHQXW-M and NHQXW-H significantly reduced fecal water content （P<0.05， P<0.01） and intestinal propulsion rate （P<0.01）， alleviated neurological deficits （P<0.01）， decreased hematoma volume （P<0.01） and Evans blue extravasation （P<0.01）， increased Claudin-5 protein expression （P<0.05， P<0.01） and fluorescence intensity （P<0.01）， and decreased PLVAP protein expression （P<0.01） and fluorescence intensity （P<0.05， P<0.01）， as well as MMP-2 （P<0.05， P<0.01） and MMP-9 （P<0.01） expression. NHQXW-H downregulated p-Akt （P<0.05）， p-ERK1/2 （P<0.05）， p-STAT3 （P<0.01）， and p-YAP （P<0.05）， with the most significant effect observed on STAT3 phosphorylation. ConclusionNHQXW effectively alleviates neurological deficits and BBB injury in ICH mice with Tanre Fushi syndrome， primarily by inhibiting STAT3 activation.

Objective: To investigate the incidence and influencing factors of falls among the elderly in Ningbo City, Zhejiang Province, so as to provide the basis for developing effective prevention strategies. Methods: The residents aged 60 years and above in Haishu District and Yuyao City of Ningbo City were selected by the multi-stage cluster random sampling method from June to October 2022. Demographic information, fall incidence in the past year, history of disease and self-rated health were collected through questionnaire surveys. Incidence of falls was descriptively analyzed, and factors affecting falls were identified using a multivariable logistic regression model stratified by gender and age. Results: A total of 1 275 elderly people were surveyed, including 635 men and 640 women. The median age was 72.00 (interquartile range, 13.00) years. In the past year, 158 residents fell, accounting for 12.39%. Additionally, 14 individuals experienced two or more falls, accounting for 8.86%. The incidence of falls was 14.69% in women, which was higher than the 10.08% in men (P<0.05). The incidence of falls was 14.86% in the elderly over 70 years, which was higher than the 9.39% in those aged 60 to 70 years (P<0.05). Multivariable logistic regression showed that the educational level (primary school and above, OR=0.501, 95%CI: 0.301-0.836), heart disease (present, OR=1.996, 95%CI: 1.076-3.703), and self-rated health status (good, OR=0.529, 95%CI: 0.319-0.875) were factors affecting falls in women; educational level (primary school and above, OR=0.514, 95%CI: 0.285-0.928) and self-rated health status (good, OR=0.456, 95%CI: 0.253-0.824) were factors affecting falls in residents aged 60 to 70 years. Conclusion Fall risk among the elderly is associated with gender, age, heart disease, educational level and self-rated health status, and the influencing factors for falls vary in different genders and ages.

Objective To evaluate the short-term clinical effect of arthroscopic repair of rotator cuff injury with all-suture anchor using a prospective and single-cohort clinical trial.Methods Twenty-five patients with rotator cuff injuries(1.5 cm0.05).Conclusion Arthroscopic rotator cuff repair with all-suture anchor is feasible and safe,and has good short-term clinical effect.

Stroke is a neurological disease characterized by cerebral ischemia or hemorrhagic injury， leading to death and disability worldwide. At present， there is still a lack of neuroprotective drugs for the treatment of stroke. Niuhuang Qingxin pills are Chinese patent medicine recommended by guidelines and expert consensus for preventing and treating stroke. Studies have shown that Niuhuang Qingxin pills have sedative， anticonvulsant， antipyretic， and other pharmacological effects. Moreover， the main components， such as flavonoids， phenolic acids， and saponins， also exhibit strong pharmacological activities which can improve stroke-induced nerve damage. Studies have confirmed that representative ingredients such as baicalin and ginsenosides can interfere with multiple pathological events， including blood-brain barrier destruction， oxidative stress， inflammatory response， apoptosis， and autophagy， and the mechanisms of the essential ingredients are related to the action on important targets such as Kelch-like ECH-associated protein 1 （Keap1）， matrix metalloproteinase-9 （MMP-9）， and high-mobility group box 1 （HMGB1）， and involvement in the regulation of Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB）， nuclear factor E₂-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）， and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathways. This article summarized the research status of Niuhuang Qingxin pills and the experimental pharmacological progress of common ingredients in the prevention and treatment of stroke to provide clues for further research into Niuhuang Qingxin pills and the development of active ingredients of traditional Chinese medicines.

Objective To evaluate the quality of prediction model on healthcare-associated infections in China,so as to standardize research process and reporting methods.Methods It performed a literature search for healthcare-associated infections prediction model studies published using the following databases by the end of 2022.After independently screening the literature and cross-checking the extracted data according to the inclusion and exclusion criteria,the research team applied the prediction model risk of bias assessment tool(PROBAST)to evaluate the methodological quality,and the transparent reporting of a multivariable prediction model for individual prognosis or diagnosis(TRIPOD)statement to evaluate the quality of study reporting.Results A total of 81 healthcare-associated infections prediction studies were identified.Their median PROBAST overall adherence were 58.11%±13.88%,median TRIPOD adherence were 56.11%±16.35%.The main methodological flaws involved participants defined,ignored complexities in data,and omitted missing data.The reporting flaws lay in the items of risk groups,sample size,and supplementary information.Conclusion There are methodological deficiencies and incomplete reporting of domestic hospital infection prediction modelling studies,which limit the reliability and applicability of the results and leave much room for improvement.

Objective To evaluate the quality of prediction model on healthcare-associated infections in China,so as to standardize research process and reporting methods.Methods It performed a literature search for healthcare-associated infections prediction model studies published using the following databases by the end of 2022.After independently screening the literature and cross-checking the extracted data according to the inclusion and exclusion criteria,the research team applied the prediction model risk of bias assessment tool(PROBAST)to evaluate the methodological quality,and the transparent reporting of a multivariable prediction model for individual prognosis or diagnosis(TRIPOD)statement to evaluate the quality of study reporting.Results A total of 81 healthcare-associated infections prediction studies were identified.Their median PROBAST overall adherence were 58.11%±13.88%,median TRIPOD adherence were 56.11%±16.35%.The main methodological flaws involved participants defined,ignored complexities in data,and omitted missing data.The reporting flaws lay in the items of risk groups,sample size,and supplementary information.Conclusion There are methodological deficiencies and incomplete reporting of domestic hospital infection prediction modelling studies,which limit the reliability and applicability of the results and leave much room for improvement.

Objective To evaluate the quality of prediction model on healthcare-associated infections in China,so as to standardize research process and reporting methods.Methods It performed a literature search for healthcare-associated infections prediction model studies published using the following databases by the end of 2022.After independently screening the literature and cross-checking the extracted data according to the inclusion and exclusion criteria,the research team applied the prediction model risk of bias assessment tool(PROBAST)to evaluate the methodological quality,and the transparent reporting of a multivariable prediction model for individual prognosis or diagnosis(TRIPOD)statement to evaluate the quality of study reporting.Results A total of 81 healthcare-associated infections prediction studies were identified.Their median PROBAST overall adherence were 58.11%±13.88%,median TRIPOD adherence were 56.11%±16.35%.The main methodological flaws involved participants defined,ignored complexities in data,and omitted missing data.The reporting flaws lay in the items of risk groups,sample size,and supplementary information.Conclusion There are methodological deficiencies and incomplete reporting of domestic hospital infection prediction modelling studies,which limit the reliability and applicability of the results and leave much room for improvement.

Objective To evaluate the quality of prediction model on healthcare-associated infections in China,so as to standardize research process and reporting methods.Methods It performed a literature search for healthcare-associated infections prediction model studies published using the following databases by the end of 2022.After independently screening the literature and cross-checking the extracted data according to the inclusion and exclusion criteria,the research team applied the prediction model risk of bias assessment tool(PROBAST)to evaluate the methodological quality,and the transparent reporting of a multivariable prediction model for individual prognosis or diagnosis(TRIPOD)statement to evaluate the quality of study reporting.Results A total of 81 healthcare-associated infections prediction studies were identified.Their median PROBAST overall adherence were 58.11%±13.88%,median TRIPOD adherence were 56.11%±16.35%.The main methodological flaws involved participants defined,ignored complexities in data,and omitted missing data.The reporting flaws lay in the items of risk groups,sample size,and supplementary information.Conclusion There are methodological deficiencies and incomplete reporting of domestic hospital infection prediction modelling studies,which limit the reliability and applicability of the results and leave much room for improvement.

Objective To evaluate the quality of prediction model on healthcare-associated infections in China,so as to standardize research process and reporting methods.Methods It performed a literature search for healthcare-associated infections prediction model studies published using the following databases by the end of 2022.After independently screening the literature and cross-checking the extracted data according to the inclusion and exclusion criteria,the research team applied the prediction model risk of bias assessment tool(PROBAST)to evaluate the methodological quality,and the transparent reporting of a multivariable prediction model for individual prognosis or diagnosis(TRIPOD)statement to evaluate the quality of study reporting.Results A total of 81 healthcare-associated infections prediction studies were identified.Their median PROBAST overall adherence were 58.11%±13.88%,median TRIPOD adherence were 56.11%±16.35%.The main methodological flaws involved participants defined,ignored complexities in data,and omitted missing data.The reporting flaws lay in the items of risk groups,sample size,and supplementary information.Conclusion There are methodological deficiencies and incomplete reporting of domestic hospital infection prediction modelling studies,which limit the reliability and applicability of the results and leave much room for improvement.