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.

Objective To assess the health risks of gaseous pollutants in the indoor air of hair and beauty salons in Jinan, and to provide technical support for strengthening the hygiene management of hair and beauty salons in Jinan and promoting the improvement of conditions. Methods Every year, indoor air samples were collected from 10-16 selected hair salons and beauty salons in Jinan, and relevant information on practitioners was also collected. According to the ^“Technical Guidelines for Environmental Health Risk Assessment of Chemicals^”, an assessment was conducted on the carcinogenic and non-carcinogenic risks of inhalation pathways of gaseous pollutants in the indoor air of hair salons and beauty salons. Results Benzene, toluene, xylene, formaldehyde, and ammonia were detected in the indoor air of hair salons and beauty salons. Formaldehyde, benzene, and ammonia all exceeded the standard in hair salons and beauty salons. The median risk values of formaldehyde and benzene for carcinogenesis in hair salons and beauty salons were both greater than 10^-6, with maximum values higher than 10^-4. The median chronic non-carcinogenic risk value of formaldehyde in the indoor air of hair salons and beauty salons was greater than 1. The median chronic non-carcinogenic risk values for benzene and ammonia were both less than 1, but the maximum risk value was greater than 1. Conclusion Benzene and formaldehyde in the indoor air of hair salons and beauty salons in Jinan City have carcinogenic and non-carcinogenic risks, while ammonia has non-carcinogenic risks, which should be paid attention to.

Moyamoya disease is a chronic cerebrovascular disease characterized by stenosis or occlusion at the terminal portion of the internal carotid artery, accompanied by the formation of an abnormal vascular network. If these cerebral angiography findings are only seen in one hemisphere of the brain, it is unilateral moyamoya disease (U-MMD). Numerous studies have shown that U-MMD is not uncommon. The latest version of diagnostic criteria has clearly diagnosed U-MMD as Moyamoya disease, rather than being a "possible" moyamoya disease. The pathogenesis of this disease involves genetic, immune, and environmental factors, but the exact cause is currently unclear. Compared with the bilateral moyamoya disease, the U-MMD has different clinical features and imaging manifestations, and has better surgical outcome, and many risk factors promote its progression to the contralateral side. This article reviews the epidemiology, pathogenesis, clinical characteristics, treatment, and surgical outcome of U-MMD, and looks forward to the future research directions.

Aim To investigate the intracellular up-take and target protein binding characteristics of two Bruton's tyrosine kinase inhibitors(BTKi)with differ-ent selectivities to provide further insights into the mechanisms of drug off-target-related bleeding risk.Methods Ibrutinib(non-selective BTKi)and za-nubrutinib(selective BTKi)were used as study drugs.After incubation of MEC-1 cells and human platelets with drugs,the cellular thermal shift assay(CETSA)was combined with Western blot to obtain the melting curve and isothermal curve to analyze the binding char-acteristics of the two drugs with the target protein BTK.After incubation of MEC-1 cells and human platelets with drugs,the concentrations of the two drugs were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS)to analyze the intracellular uptake of the two drugs.Results CETSA analysis confirmed that zanubrutinib was more selective for the target protein BTK compared to ibrutinib.LC-MS/MS analysis showed that both drugs were uptaken intracel-lularly by MEC-1 cells and platelets in a concentration-dependent manner.Conclusions While BTKi targe-ting BTK to B lymphocytes exerts therapeutic effects,off-target effects on platelets due to differences in their intracellular uptake,and target-binding characteristics may be one of the reasons for the differences in bleed-ing risk across selective BTKi.

AIM To investigate the renal protective effects of Shiwei Ruxiang Powder on gouty nephritis in rats based on mitophagy.METHODS Rats were randomly divided into the blank group,the model group,the low-dose,medium-dose,and high-dose Shiwei Ruxiang Powder groups(200,400,800 mg/kg)and allopurinol group(10 mg/kg).The rat model of gouty nephropathy was established by gavage of potassium oxyzinate(750 mg/kg)and uric acid(300 mg/kg).The rats had their levels of UA,SCr,BUN,XOD,SOD,MDA,ROS measured by automatic biochemical analyzer,ELISA and chemical fluorescence method;their renal pathological changes observed by HE staining;their apoptosis of renal tissue cells observed by TUNEL staining;and their mRNA and protein expressions of IL-1β,TNF-α,Bax,Bcl-2,caspase-3,caspase-9,PINK1,Parkin and LC3-Ⅱ detected by RT-qPCR and Western blot.RESULTS Compared with the model group,Shiwei Ruxiang Powder groups displayed dose-dependently decreased serum levels of UA,BUN and SCr,renal deposition of urate crystal and apoptosis(P<0.05);decreased renal levels of ROS and inflammatory factors IL-1β and TNF-α(P<0.05);and increased renal expressions of mitochondrial autophagy-related proteins PINK1,Parkin and LC3-Ⅱ(P<0.01).CONCLUSION Shiwei Ruxiang Powder may relieve gouty kidney injury in rats by reducing the uric acid level,the renal oxidative stress and inflammatory response,and activating mitophagy pathway as well.

Neck dissection(ND)is one of the main treatment methods for oral and maxillofacial malignancies.Although ND type is in con-stant improvement,but intraoperative peal-pull-push injury of the accessory nerve,muscle,muscle membrane,fascia and ligament induced shoulder syndrome(SS)is still a common postoperative complication,combined with the influence of radiochemotherapy,not only can cause pain,stiffness,numbness,limited dysfunction of shoulder neck and arm,but also may have serious impact on patient's life quality and phys-ical and mental health.At present,there is still a lack of a systematic evaluation and rehabilitation management program for postoperative SS of oral and maxillofacial malignant tumors.Based on the previous clinical practice and the current available evidence,refer to the relevant lit-erature at home and abroad,the experts in the field of maxillofacial tumor surgery and rehabilitation were invited to discuss,modify and reach a consenusus on the etiology,assessment diagnosis,differential diagnosis,rehabilitation strategy and prevention of SS,in order to provide clinical reference.

Objective:To investigate the therapeutic efficacy and underlying mechanisms of the traditional Chinese medicine formula "Hua Xian Fang" (HXF) in the treatment of radiation-induced pulmonary fibrosis (RIPF).Methods:In vivo experiment, 36 male specific pathogen free (SPF)-grade C57BL/6 mice aged 6-8 weeks were randomly divided into the control, irradiation (17 Gy thoracic irradiation), and irradiation+HXF groups (17 Gy thoracic irradiation+HXF). After 16 weeks, lung coefficient, HE staining and Masson staining were used to evaluate the degree of pulmonary inflammation and fibrosis. Immunohistochemistry was performed to measure the expression levels of α-smooth muscle actin (α-SMA) in lung tissues. Quantitative real-time PCR (qPCR) was performed to detect the mRNA expression levels of interferon-γ (IFN-γ). Enzyme linked immunosorbent assay (ELISA) was conducted to determine the levels of IFN-γ in serum and bronchoalveolar lavage fluid (BALF). During in vitro experiment, NIH/3T3 fibroblasts were stimulated with IFN-γ after 6 Gy irradiation, followed by 48 hours of culture. qPCR, immunofluorescence staining, and Western blot were used to assess the expression of α-SMA and collagen Ⅰ at the transcription and protein levels. One way ANOVA was used for multiple group comparisons, and Tukey test was used for inter group multiple comparisons. Results:Compared to the control group, mice in the irradiation group showed significant increases in lung coefficient, Szapiel score, Ashcroft score, and α-SMA expression in lung tissues (all P<0.001). Compared to the irradiation group, the irradiation+HXF group exhibited significant decreases in the above indicators (all P<0.001). qPCR demonstrated that the mRNA expression levels of IFN-γ were significantly higher in the irradiation+HXF group than that in the irradiation group ( P=0.001). ELISA results showed that the levels of IFN-γ in serum and BALF were significantly elevated in the irradiation+HXF group compared to those in the irradiation group ( P=0.032, 0.037). In vitro experiment revealed that after irradiation, the expression levels of α-SMA and collagen Ⅰ mRNA and protein in NIH/3T3 cells were significantly increased, while decreased after IFN-γ stimulation. Conclusion:HXF effectively alleviates RIPF, probably by the upregulation of IFN-γ in blood and tissues and inhibition of fibroblast activation.

Objective To explore the dynamic changes and mechanisms of neurological and cognitive functions in mice with traumatic brain injury (TBI). Methods Totally 60 12⁃month⁃old Balb/ c mice were divided into control group (10 in group) and TBI group (50 in group). TBT model mice were divided into 5 subgroups according to the time of model construction, including model 1 day, model 1 day, model 3 day, model 7 day, model 14 days and model 28 days group with 10 in each group. At the 29th day of the experiment, neurological scores and step down tests were carried out. After the test, the mice were sacrificed for brains which were detected by immunohistochemistry staining, inflammatory cytokine tests and Western blotting. Results Compared with the control group, the neurological scores of mice in TBI group increased, and then decreased after the 7th day when the scores reached the peak. However, the latency of step down errors was lower than control group, and the number of step down errors was higher than control group which had no changes. Compared with the control group, the expression of lonized calcium⁃binding adapter molecule 1(IBA1), chemokine C⁃X3⁃C⁃motif ligand1 (CX3CL1), C⁃X3⁃C chemokine receptor 1(CX3CR1), NOD⁃like receptor thermal protein domain associated protein 3 (NLRP3), and phosphorylation nuclear factor(p⁃NF)⁃κB in TBI group increased and reached to the peak at the 7th day, and then started to decrease. At the same time, the levels of inflammatory cytokines interleukin⁃6(IL⁃6) and tumor necrosis factor⁃α(TNF⁃α) first increased to the peak, and then began to decrease. However, compared with the control group, the expression of amyloid β(Aβ) protein and p⁃Tau protein in the model group continued to increase at all time. Conclusion The TBI model caused continuous activation of microglia along with inflammatory response, which first increased and then decreased, resultsing in neurological scores changes. In addition, the inflammatory response may act as a promoter of Aβ protein deposition and Tau protein phosphorylation, leading to cognitive impairment in mice.

Surgical operation is the main treatment of oral and maxillofacial tumors.Dysphagia is a common postoperative complication.Swal-lowing disorder can not only lead to mis-aspiration,malnutrition,aspiration pneumonia and other serious consequences,but also may cause psychological problems and social communication barriers,affecting the quality of life of the patients.At present,there is no systematic evalua-tion and rehabilitation management plan for the problem of swallowing disorder after oral and maxillofacial tumor surgery in China.Combining the characteristics of postoperative swallowing disorder in patients with oral and maxillofacial tumors,summarizing the clinical experience of ex-perts in the field of tumor and rehabilitation,reviewing and summarizing relevant literature at home and abroad,and through joint discussion and modification,a group of national experts reached this consensus including the core contents of the screening of swallowing disorders,the phased assessment of prognosis and complications,and the implementation plan of comprehensive management such as nutrition management,respiratory management,swallowing function recovery,psychology and nursing during rehabilitation treatment,in order to improve the evalua-tion and rehabilitation of swallowing disorder after oral and maxillofacial tumor surgery in clinic.

Cryoablation therapy with explicit anti-tumor mechanisms and histopathological manifestations has a long history.A large number of clinical practice has shown that cryoablation therapy is safe and effective,making it an ideal tumor treatment method in theory.Previously,its efficacy and clinical application were constrained by the limitations of refrigerants and refrigeration equipment.With the development of the new generation of cryoablation equipment represented by argon helium knives,significant progress has been made in refrigeration efficien-cy,ablation range,and precise temperature measurement,greatly promoting the progression of tumor cryoablation technology.This consensus systematically summarizes the mechanism of cryoablation technology,indications for oral mucosal melanoma(OMM)cryotherapy,clinical treatment process,adverse reactions and management,cryotherapy combination therapy,etc.,aiming to provide reference for carrying out the standardized cryoablation therapy of OMM.