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.

AIM To establish a quantitative analysis of multi-components by single-marker(QAMS)method for the simultaneous content determination of gastrodin,parishin E,syringin,parishin B,parishin C,ferulic acid,parishin A,buddleoside,harpagoside and cinnamic acid in Tianma Toufengling Capsules.METHODS The analysis was performed on a 30℃thermostatic GL Science InertsilTM ODS-3 column(150 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,280 nm.Syringin was used as an internal standard to calculate the relative correction factors of the other nine constituents,after which the content determination was made.RESULTS Ten constituents showed good linear relationships within their own ranges(r≥0.999 7),whose average recoveries were 98.53%-102.22%with the RSDs of 1.26%-2.68%.The result obtained by QAMS approximated those obtained by external standard method.CONCLUSION This accurate and specific method can be used for the quality control of Tianma Toufengling Capsules.

Objective: To explore the relationship between processed food consumption and blood pressure level of students in a university in Yunnan Province, so as to provide the reference for preventing hypertension in university students. Methods: In October 2021, a cluster sampling method was used to select 4 781 freshmen from a university in Kunming, Yunnan Province. The frequency of processed food consumption of university students was assessed by using the dietary frequency questionnaire, and height, weight and blood pressure were measured. Mann-Whitney test and Kruskal-Wallis test were used to compare the differences in blood pressure level of university students with different demographic variables, and the association between processed food consumption and blood pressure level was analyzed with a generalized linear model. Results: Among the students of a university in Yunnan Province, the detection rates of systolic prehypertension and hypertension were 33.86% and 1.23%, and the detection rates of diastolic prehypertension were 32.13% and hypertension 7.22%. The results of generalized linear model analysis showed that after controlling for demographic variables and other variables that might affect the blood pressure level of university students, the consumption of processed food (bread and cake: β =0.15, 95% CI =0.01-0.29) and ultra processed food (coffee beverage: β =-0.29, 95% CI =-0.54--0.03) were associated with systolic blood pressure level( P <0.05). The consumption of processed food (salted duck egg: β =0.21, 95% CI =0.01-0.41) was correlated with the diastolic blood pressure of college students ( P <0.05). Conclusions Processed food consumption in university students may increase the risk of high blood pressure.The education of healthy eating among college students should be strengthened to reduce the consumption of processed foods.

ObjectiveTo investigate the effect of 8-week aerobic exercise on the improvement of core behaviors of male and female autistic mice induced by valproic acid (VPA). MethodsExperimental mice were randomly divided into the control group (CTL), VPA-induced autism group (VPA) and VPA+aerobic exercise group (VEX), with 10 mice in each group. The pregnant mice were injected with VPA intraperitoneally at E12.5, and their offspring were used as autistic mice. Pups were weaned 28 d after birth and began an 8-week aerobic exercise intervention. The day after exercise, mice were tested in behavioral experiments to detect exploratory behavior, social skills, repetitive stereotypic behavior, cognitive ability and mood. The mice were tested for social skills, repetitive stereotyped behaviors, cognitive and learning memory abilities, exploratory behaviors, and emotions by behavioral assays on the following day after the exercise. ResultsBoth male and female mice in the CTL group showed a significant decrease in the total distance and percentage of time spent in the interaction zone in the 4th socialization compared to the 1st socialization (P<0.01); the total distance and percentage of time spent in the interaction zone in the 5th socialization was significantly increased compared to the 4th socialization (P<0.01); in VPA group, both male and female mice showed no significant change in the total distance and percentage of time spent in the interaction zone in the 4th and 5th socialization; in the VEX group, the total distance and percentage of time spent in the interaction zone by male mice in the 4th socialization was significantly decreased compared to the 1st socialization (P<0.01, P<0.05); and in the VEX group the total distance and percentage of time spent in the social interaction zone by both male and female mice in the 5th socialization was significantly increased compared to the 4th socialization (P<0.01, P<0.05). The results of the first phase of three-box socialization experiment showed that male and female mice in the CTL group spent more time socializing with their social partners than in contact with the empty cages (P<0.01); there was no difference in the time spent by male and female mice in the VPA group in socializing with their social partners and the empty cages; and male and female mice in the VEX group spent a longer time socializing with their social partners(P<0.01). The results of the second phase of three-box test showed that male and female mice in the CTL group showed a significant tendency to socialize with new social partners (P<0.01), whereas no significant changes were observed in the mice of VPA group; aerobic exercise significantly ameliorated this deficit in male and female mice with autism. Compared with the CTL group, VPA-induced significant decreases were observed in the total distance freely moved in the central area of the open field, the time and percentage of time spent in the open arm of cross maze, and the distance and time spent in the white box in both male and female autistic mice (P<0.01); a significant increase in the number of bead burials and time spent in self-grooming (P<0.01); a significant decrease in the cognitive index (P<0.01); a significantly longer latency to find the platform, and significantly decreased the percentage of time spent in the target quadrant and the number of times they traversed the platform (P<0.01). Compared with the VPA group, after 8 weeks of aerobic intervention, male and female mice in the VEX group showed a significant increase in total distance, open-arm dwell time, and percentage of free movement in the central area of the empty field (P<0. 05), and a trend toward a decrease in the dwell time of females in the white box was not significant, the number of beads burying and the time of self-combing were significantly lower(P<0.01, P<0.05); and a significant increase in cognitive index (P<0.05), a significantly shorter time to find the platform, and significantly increased percentage of time spent in the target quadrant and the number of times they traversed the platform (P<0.01), showing excellent learning memory. ConclusionAutistic mice severely suffer from social and cognitive impairments, repetitive stereotyped behaviors, decreased activity level, and the exhibition of anxiety. 8 weeks of aerobic exercise can improve the social and cognitive abilities, alleviate the stereotyped repetitive behaviors, increase the activity level, and positively regulate the anxiety in autistic mice. It is hypothesized that aerobic exercise has an important role in motor rehabilitation of autism, in order to provide a theoretical basis for clinical research.

italic>Astragalus polysaccharides are the most immunoregulatory active and abundant substances in Astragalus, with anti-tumor, anti-viral, and immune-promoting biological activities. They have been widely used in clinical practice. Previous studies have found that Astragalus polysaccharides are mainly composed of two different polysaccharides, APS-I (>2 000 kDa) and APS-Ⅱ (10 kDa), with APS-Ⅱ (10 kDa) being the most active component of Astragalus polysaccharides. We used α-1,4-glucan endo-hydrolysis enzyme to degrade APS-Ⅱ into oligosaccharides, and screened the immune activity of oligosaccharides in vitro. We found that the overall immune activity of 2-9 oligosaccharides was low, while the immune activity of 10-14 oligosaccharides was strong, and the activity was better than that of untreated APS-Ⅱ. To investigate the key structural features of APS-Ⅱ oligosaccharides that play a role in immune activity, we used MALDI-TOF-MS biochemical mass spectrometry and high-resolution mass spectrometry instrument ESI-Q Exactive-MS to analyze the APS enzymatic oligosaccharides. By comparing, we found that 10-14 oligosaccharides contain 1→4 and 1→6 branched structures with coexisting linkages, suggesting that 1→4 and 1→6 linkages in branched structures are key structural features that play a role in the immune activity of APS-Ⅱ, laying a theoretical foundation for the structure-activity relationship of Astragalus polysaccharides and oligosaccharides.

Objective Little is known about the association between whole-blood nicotinamide adenine dinucleotide(NAD+)levels and nabothian cysts.This study aimed to assess the association between NAD+levels and nabothian cysts in healthy Chinese women. Methods Multivariate logistic regression analysis was performed to analyze the association between NAD+levels and nabothian cysts. Results The mean age was 43.0±11.5 years,and the mean level of NAD+was 31.3±5.3 μmol/L.Nabothian cysts occurred in 184(27.7%)participants,with single and multiple cysts in 100(15.0%)and 84(12.6%)participants,respectively.The total nabothian cyst prevalence gradually decreased from 37.4%to 21.6%from Q1 to Q4 of NAD+and the prevalence of single and multiple nabothian cysts also decreased across the NAD+quartiles.As compared with the highest NAD+quartile(≥34.4 μmol/L),the adjusted odds ratios with 95%confidence interval of the NAD+Q1 was 1.89(1.14-3.14)for total nabothian cysts.The risk of total and single nabothian cysts linearly decreased with increasing NAD+levels,while the risk of multiple nabothian cysts decreased more rapidly at NAD+levels of 28.0 to 35.0 μmol/L. Conclusion:Low NAD+levels were associated with an increased risk of total and multiple nabothian cysts.

Background and objective Lung cancer is the malignant tumor with the highest incidence rate and the heaviest disease burden in China.In recent years,lung cancer has shown a high incidence trend,seriously affecting the health of the population.In this paper,we analyze the characteristics of lung cancer incidence in 2019 and the trend of incidence rate from 2010-2019 in the tumor registration area of Gansu province,in order to provide a reference basis for the development of lung cancer prevention and control strategies in Gansu province.Methods By analyzing the cases of lung cancer incidence in the tumor registration area of Gansu province in 2019,we calculated the incidence rate,medium incidence rate,world in-cidence rate and other related indexes;we used Joinpoint to calculate the annual percentage change(APC)for trend analysis.Results In 2019,a total of 3757 new cases of lung cancer were reported in Gansu province,accounting for 14.96%of all new malignant tumors.The incidence rate,medium incidence rate and world incidence rate and world rate of lung cancer were 40.52/105,25.78/105,25.86/105;and the cumulative rate of 0-74 years old,and the truncation rate of 35-64 years old were 3.23%,40.03/105,respectively.The incidence of lung cancer rises with age,and is high in the age group of 40 years and above,and the incidence peaks in the male and female populations in the group of 75 years and above,and the group of 80 years and above,respectively.The crude incidence rate of lung cancer in the tumor registration area of Gansu province from 2010-2019 showed an overall increasing trend,and the rate of increase was relatively fast,with an APC 5.39%(P<0.05);Separately,accord-ing to gender,urban and rural areas,the incidence of lung cancer in all populations showed an increasing trend,and the APC of male,female,urban and rural populations were 4.98%,6.39%,6.26%,and 4.64%,respectively(all P<0.05).According to the trend analysis of lung cancer incidence rate by age group,only lung cancer incidence in the age group of 65 years and above increased at an annual average rate of 4.15%(P<0.05).Conclusion The incidence rate of lung cancer in the tumor registration area of Gansu province from 2010 to 2019 shows a rising trend year by year,and there are differences in the incidence of lung cancer in people of different genders,regions and age groups,so comprehensive prevention and control work should be carried out for the key populations of lung cancer incidence.

Objective:To explore the efficacy of immune checkpoint inhibitors combined with adjuvant chemotherapy in patients with phase III gastric cancer and esophagogastric junction cancer.Methods:This study used a retrospective cohort study method based on real-world data. Clinical data of 403 patients with stage III gastric/esophagogastric junction cancer who underwent gastrectomy followed by adjuvant therapy in the Department of Gastric Surgery at Sun Yat-sen University Cancer Center from January 2020 to December 2023 were retrospectively collected. The study cohort comprised 147 (36.5%) patients with stage IIIA, 130 (32.3%) with stage IIIB, and 126 (31.3%) with stage IIIC gastric/esophagogastric junction cancer. Of them, 15 (3.7%) were HER-2 positive, 25 (6.2%) dMMR, and 22 (5.5%) patients Epstein-Barr virus encoding RNA (EBER) positive. Based on treatment plans, the patients were divided into immune checkpoint inhibitor combined with chemotherapy group (immune therapy group, n=110, 71 males and 39 females, median age 59 years old) and chemotherapy alone group (chemotherapy group, n=293, 186 males and 107 females, median age 60 years old). All patients in the immunotherapy group received immune checkpoint inhibitors targeting the programmed cell death protein-1 (PD-1) and its ligand (PD-L1). Of them, 85 received pembrolizumab, 10 received sintilimab, 8 received tislelizumab, 4 received camrelizumab, 2 received toripalimab, and 1 received pabocizumab. The adjuvant chemotherapy regimens used among the chemotherapy alone group includes SOX regimen (132 cases), XELOX (102 cases), S-1 monotherapy (44 cases), and other regimens (15 cases). The 3-year DFS rate of the two groups was compared, and subgroup analysis was conducted based on different ages, molecular phenotypes, pTNM staging, extranodal infiltration, and tumor length. Results:The median follow-up was 20.5 months (range 3.1~46.3), with a 3-year overall DFS rate of 61.4% for the entire 403 patients. The 3-year DFS rate for the immunotherapy group was 82.7%, higher than the chemotherapy alone group (58.8%), with a statistically significant difference ( P=0.021). Multivariate analysis showed that postoperative immunotherapy was a protective factor for DFS (HR=0.352, 95%CI: 0.180~0.685). Subgroup analysis showed that stage IIIC (HR=0.416, 95%CI: 0.184~0.940), aged ≥60 years (HR=0.336, 95%CI: 0.121~0.934) and extranodal invasion (HR=0.378, 95%CI: 0.170~0.839) were associated with benefit from the combined immune adjuvant chemotherapy, while no association was observed for MMR, HER-2 or EBER status. Conclusion:Stage III gastric/esophagogastric junction cancer patients may benefite from postoperative immune checkpoint inhibitor combined with adjuvant chemotherapy in real-world settings.

Background and aim:Hepatic ischemia-reperfusion injury(IRI)is a significant challenge in liver trans-plantation,trauma,hypovolemic shock,and hepatectomy,with limited effective interventions available.This study aimed to investigate the role of leukocyte cell-derived chemotaxin 2(LECT2)in hepatic IRI and assess the therapeutic potential of Lect2-short hairpin RNA(shRNA)delivered through adeno-associated virus(AAV)vectors. Materials and methods:This study analyzed human liver and serum samples from five patients under-going the Pringle maneuver.Lect2-knockout and C57BL/6J mice were used.Hepatic IRI was induced by clamping the hepatic pedicle.Treatments included recombinant human LECT2(rLECT2)and AAV-Lect2-shRNA.LECT2 expression levels and serum biomarkers including alanine aminotransferase(ALT),aspartate aminotransferase(AST),creatinine,and blood urea nitrogen(BUN)were measured.Histological analysis of liver necrosis and quantitative reverse-transcription polymerase chain reaction were performed. Results:Serum and liver LECT2 levels were elevated during hepatic IRI.Serum LECT2 protein and mRNA levels increased post reperfusion.Lect2-knockout mice had reduced weight loss;hepatic necrosis;and serum ALT,AST,creatinine,and BUN levels.rLECT2 treatment exacerbated weight loss,hepatic necrosis,and serum biomarkers(ALT,AST,creatinine,and BUN).AAV-Lect2-shRNA treatment significantly reduced weight loss,hepatic necrosis,and serum biomarkers(ALT,AST,creatinine,and BUN),indicating thera-peutic potential. Conclusions:Elevated LECT2 levels during hepatic IRI increased liver damage.Genetic knockout or shRNA-mediated knockdown of Lect2 reduced liver damage,indicating its therapeutic potential.AAV-mediated Lect2-shRNA delivery mitigated hepatic IRI,offering a potential new treatment strategy to enhance clinical outcomes for patients undergoing liver-related surgeries or trauma.

Objective To investigate the correlation between traditional Chinese medicine(TCM)syndrome elements and risk factors in children with IgA vasculitis(IgAV,also known as Henoch-Sch?nlein purpura).Methods The medical records of 131 children with IgAV were retrospectively analyzed.And then the distribution of their TCM syndrome elements was investigated,and the correlation of TCM syndrome elements with the gender,age,clinical symptoms,as well as risk factors such as mosquito bite,pathogen infection,and allergic rhinitis was analyzed.Results(1)Among the 131 children with IgAV,the diseases-location syndrome elements of IgAV involved lung in 97 cases(74.05%),spleen in 61 cases(46.56%),kidney in 54 cases(41.22%),liver in 17 cases(12.98%),and heart in 11 cases(8.40%);the disease-nature syndrome elements of IgAV involved blood stasis in 131 cases(100.00%),wind-damp in 125 cases(95.42%),wind-heat in 90 cases(68.70%),damp-heat in 72 cases(54.96%),blood heat in 49 cases(37.40%),qi deficiency in 19 cases(14.50%),and yin deficiency in three cases(2.29%).(2)There were 69 cases(52.67%)of females and 62 cases(47.33%)of males among the IgAV children,with females outnumbering males.The age group of IgAV children was predominated by five to six years old,and 10 cases(7.63%)were younger than four years old,18 cases(13.74%)were four years old,39 cases(29.77%)were five years old,34 cases(25.95%)were six years old,17 cases(12.98%)were seven years old,and 13 cases(9.92%)were older than seven years old.The disease-nature syndrome elements such as blood stasis,wind-damp,wind-heat,and damp-heat were frequently seen in the age group of five to seven years old,yin deficiency was frequently seen in the age group older than seven years,and blood stasis was seen in all age groups.(3)The results of logistic regression analysis of the correlation between TCM syndrome elements and risk factors in IgAV patients showed that allergic rhinitis was positively correlated with blood stasis[OR=2.236,95%CI(1.049-4.007)],damp-heat[OR=2.183,95%CI(1.554-3.671)]and wind-damp[OR=1.202,95%CI(1.050-2.409)];pathogen infection was positively correlated with blood stasis[OR=3.199,95%CI(1.457-4.101)]and damp-heat[OR=1.119,95%CI(1.072-2.009)];mosquito bite was positively correlated with blood stasis[OR=4.533,95%CI(1.029-9.022)]and damp-heat[OR=2.257,95%CI(1.081-13.207)];the gender was positively correlated with blood stasis[OR=1.352,95%CI(1.271-3.018)]and wind-damp[OR=1.149,95%CI(1.071-3.102)].The differences were all statistically significant(P＜0.05 or P＜0.01).Conclusion IgAV usually involves the lungs and is also related to the five zang organs.Its pathogenesis is characterized by excess in superficiality such as blood stasis and wind-damp-heat in the early stage,and is predominated by deficiency in origin such as qi deficiency and yin deficiency in the later stage.For the children with IgAV,mosquito bite,pathogen infection and allergic rhinitis are more likely to induce blood stasis and wind-damp-heat;TCM syndrome elements such as wind-heat,damp-heat,blood heat,and qi deficiency are frequently seen in the males,while TCM syndrome elements such as blood stasis,wind-damp,and yin deficiency are frequently seen in the females.