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 investigate the impact of corneal fluorescein sodium(NaF)staining on the examination results of iTrace visual function analyzer(iTrace).METHODS: Prospective cohort study. Totally 100 patients(100 eyes)with ametropia who visited the outpatient department of Anhui Eye Hospital from April to November 2024 were recruited. They were divided into an experimental group and a control group, with 50 patients(50 eyes, and only the right eyes were selected for inclusion)in each group. In the experimental group, corneal staining was performed using fluorescein sodium staining test strips, while in the control group, 1 drop of 0.9% normal saline was instilled into the eyes. The iTrace examination was conducted before the intervention and at 5, 10, and 20 min after the intervention. The total corneal higher-order aberrations, spherical aberration, coma aberration, trefoil aberration, best sphere value(RO value), asphericity factor(Q value), and corneal vertical refractive power difference(IS value)at each time of examination were recorded and compared.RESULTS: There was no statistically significant difference in the baseline levels between the two groups(all P>0.05). Intra-group comparison revealed that the total higher-order aberrations, spherical aberration, coma aberration, and trefoil aberration measured 5 min after NaF staining in the experimental group were significantly increased compared with those before staining(all P<0.05). Inter-group comparison showed that the changes(differences from the baseline)in the total corneal higher-order aberrations, spherical aberration, coma aberration, and trefoil aberration measured by iTrace 5 min after the intervention in the experimental group were significantly greater than those in the control group(all P<0.05). There was no statistically significant difference in the changes(differences from the baseline)of various iTrace parameters measured at 10 and 20 min after the intervention between the two groups(all P>0.05). There was no statistical significance in the RO value, Q value, and IS value in the two groups(all P>0.05).CONCLUSION: Corneal NaF staining can cause a short-term increase in the wavefront aberration values(total corneal higher-order aberrations, spherical aberration, coma aberration, trefoil aberration)measured by iTrace, and it gradually disappears with the passage of time. However, it has no impact on the measurement of corneal topography parameters(RO value, Q value, IS value).

As the number of patients with compromised immune function increases and fungal resistance develops, so does the risk of contracting deadly fungi in humans. Both fungi and humans are eukaryotes, so identifying unique targets for antifungal drug development is difficult. In addition, the existing antifungal drugs are limited by toxicity, drug interaction and drug resistance in practical application, which leads to the increasing incidence and fatal rate of fungal infections. Therefore, it is urgent to develop new antifungal drugs. The semi-synthetic technology using microbial fermentation products from natural sources as lead compounds has become the most used method in structural modification of antifungal drugs due to its advantages of few reaction steps and easy operation. This paper will introduce the current status of natural antifungal drugs in clinical use, as well as the latest progress in the research and development of new semi-synthetic antifungal drugs, and summarize their mechanism of action, structural modifications, advantages and disadvantages, so as to provide reference for the subsequent development of new antifungal drugs.

Aim To investigate the regulatory role and mechanism of resveratrol in inhibiting autophagy and promoting apoptosis in choroidal melanoma cells. Methods Choroidal melanoma cells (MUM2B) were divided into control and experimental groups, and treated with different concentrations of resveratrol (0, 10, 20,40,60,80 μmol ·L

Aim To explore the effect of oxaliplatin combined with epidermal growth factor receptor tyrosine kinase inhibitor AG1478 on autophagy in non-small cell lung cancer H1975 cells. Methods H1975 cells were cultured in vitro using gradient concentrations of AG1478 (0, 5, 10, 15, 20, 25, 30, 35, 40 jjimol • IT

Objective:To analyze the clinical and radiographic effectiveness of a calcium silicate-based bioactive ceramic iRoot BP Plus? pulpotomy of immature permanent teeth with complicated crown fracture and to evaluate the factors influencing its long-term success rate.Methods:The digital medical records of patients under 13 years old who had undergone iRoot BP Plus? pulpotomy in the Department of Oral Emergency or the First Clinical Division,Peking University School and Hospital of Stomatology from March 2017 to September 2022 due to complicated crown fracture of anterior teeth,and had taken at least one post-operation apical radiograph were reviewed.The clinical and radiographic information at the initial examination and follow-up period were obtained,including crown color,mobility,percussion,cold test(partial pulpotomy teeth),dental restoration,fistula,swelling or inflammation of the gingival tissue,the formation of apical foramen,pathologic radiolucency and calcification of pulp chamber or root canal obliteration.Data were tested by Fisher exact test and a multiple comparison.Results:In the study,64 patients including 37 males(57.8％)and 27 females(42.2％)with a mean age of 9.1 years were finally enrolled.The total number of permanent teeth that received pulpotomy was 75,and the average follow-up time was 19.3 months.The success rate was 93.1％with the time interval between dental injury and treatment in 24 h,while the success rate dropped to 88.2％with the time intervals beyond 24 h.The time intervals did not significantly affect the pulp survival rate(P=0.61)after pulpotomy(partial or co-ronal).The success rate 6 months after pulpotomy was 96.0％,and one-year success rate was 94.7％.A total of 23 cases were reviewed for more than 2 years after pulpotomy,and 6 cases failed.The mobility had no significant effect on the success rate(P=0.28).Pulp chamber calcification and pulp canal obli-teration were not observed in all the post-operative radiographs.Conclusion:The one year clinical and radiographic success rates obtained in this study indicate that iRoot BP Plus? is an appropriate pulp cap-ping material option for pulpotomy treatment of complicated crown fracture in immature permanent teeth without displacement injuries.This technique has broad promotional value.

Objective To investigate the effect of glycyrrhizic acid(GA)on the inflammatory and fibrotic factors in high glucose-induced glomerular mesangial cells(SV40 MES13).Methods Cultured mouse SV40 MES13 were divided into normal group(NG,5.6 mmol/L glucose),high glucose group(30 mmol/L glucose)and HG+GA group(30 mmol/L glucose+200 μmol/L GA).The expression levels of inflammatory cytokines interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6,IL-8 and α-smooth muscle actin(α-SMA)in different groups were detected by Western blotting.The fluorescence intensity of IL-1β,TNF-α and α-SMA in different groups were detected by immunofluorescence.The levels of IL-1β,TNF-α,IL-6 and IL-8 in the culture supernatant of different populations were detected by enzyme-linked immunosorbent assay(ELISA).Results The protein expressions of IL-1β,TNF-α,IL-6,IL-8 and α-SMA in HG group were significantly higher than those in NG group(P<0.01);Compared with HG group,the protein expression levels of IL-1β,TNF-α,IL-6,IL-8 and α-SMA decreased significantly in HG+GA group(P<0.05).The fluorescence intensity of inflammatory cytokines IL-1β,TNF-α and α-SMA increased in HG group than those in NG group(P<0.05);While compared with the HG group,the fluorescence intensity of IL-1β,TNF-α and α-SMA in HG+GA group decreased markedly(P<0.05).The experimental results of ELISA showed that compared with NG group,the levels of IL-1β,IL-6,TNF-α and IL-8 in cell supernatent increased in HG group(P<0.01);while the levels of IL-1β,TNF-α,IL-6,IL-8 in HG+GA group significantly lower than those in HG group(P<0.05).Conclusion Glycyrrhizic acid has certain inhibitory effect on high glucose-induced inflammatory factors and fibrotic factors in glomerular mesangial cells,which may play an important role in prevention of diabetic nephropathy.

Objective To promote the construction of a national party building indicator system for public hospitals,provide reference for the development of system documents such as quality evaluation methods for party building work in public hospitals.Methods Conduct a comprehensive and systematic collection,organization,and analysis of policy documents related to party building in public hospitals in China since 2018,and organize expert discussions.Results The positive coefficients of the three rounds of expert discussions were all 100％,and the expert authority coefficient was 0.878.A party building indicator system was constructed with 7 primary indicators and 40 secondary indicators,including leadership system and decision-making mechanism,leadership team and cadre talent team construction,grassroots party organization construction,party member team construction,medical ethics and clean governance construction,party building work guarantee,and others.Conclusion The indicator system has been unanimously recognized by experts and is authoritative and feasible,laying the foundation for the refined party building work in public hospitals.

Objective:To investigate the current situation of the use of transjugular intrahepatic portosystemic shunt (TIPS) for portal hypertension, which should aid the development of TIPS in China.Methods:The China Portal Hypertension Alliance (CHESS) initiated this study that comprehensively investigated the basic situation of TIPS for portal hypertension in China through network research. The survey included the following: the number of surgical cases, main indications, the development of Early-TIPS, TIPS for portal vein cavernous transformation, collateral circulation embolization, intraoperative portal pressure gradient measurement, commonly used stent types, conventional anticoagulation and time, postoperative follow-up, obstacles, and the application of domestic instruments.Results:According to the survey, a total of 13 527 TIPS operations were carried out in 545 hospitals participating in the survey in 2021, and 94.1% of the hospital had the habit of routine follow-up after TIPS. Most hospitals believed that the main indications of TIPS were the control of acute bleeding (42.6%) and the prevention of rebleeding (40.7%). 48.1% of the teams carried out early or priority TIPS, 53.0% of the teams carried out TIPS for the cavernous transformation of the portal vein, and 81.0% chose routine embolization of collateral circulation during operation. Most of them used coils and biological glue as embolic materials, and 78.5% of the team routinely performed intraoperative portal pressure gradient measurements. In selecting TIPS stents, 57.1% of the hospitals woulel choose Viator-specific stents, 57.2% woulel choose conventional anticoagulation after TIPS, and the duration of anticoagulation was between 3-6 months (55.4%). The limitation of TIPS surgery was mainly due to cost (72.3%) and insufficient understanding of doctors in related departments (77.4%). Most teams accepted the domestic instruments used in TIPS (92.7%).Conclusions:This survey shows that TIPS treatment is an essential part of treating portal hypertension in China. The total number of TIPS cases is far from that of patients with portal hypertension. In the future, it is still necessary to popularize TIPS technology and further standardize surgical indications, routine operations, and instrument application.

Objective To investigate the effects of Helicobacter pylori(Hp)on the proliferation,migration,apoptosis and inflammatory response of human umbilical vein endothelial cells(HUVEC)through activation of STAT3/nuclear factor κB(NF-κB)pathway.Methods HUVEC were divided into control group(without Hp infection)and Hp group(multiplicity of infection=25).Cell morphology was observed with inverted microscopy,proliferation was detected by CCK-8 assay and plate cloning assay,and the migration ability was examined by Transwell migration as-say and wound healing assay.Flow cytometry was used to detect the apoptotic rate.Real-time fluo-rescence quantitative PCR was employed to measure the mRNA expression of cytotoxin-associat-ed gene A(CagA),IL-6,IL-8,IL-1β and TNF-α.Western blotting was applied to determine the protein expression of Cyclin D1,proto-oncogene C-Myc,MMP-2,MMP-9,PCNA,Bax,Bcl-2 and STAT3/NF-κB signaling pathway.Results Hp infection resulted in suppressed proliferation and migration abilities,decreased protein levels of Cyclin D1,PCNA,C-Myc,MMP-2,MMP-9 and Bcl-2,elevated protein levels of Bax,p-STAT3/STAT3,p-NF-KB p65/NF-κB p65,raised apoptotic rate,and significantly increased mRNA levels of IL-6,IL-8,IL-1β and TNF-α(2.71±0.05 vs 1.06±0.41,1.42±0.02 vs 0.92±0.11,2.50±0.29 vs 1.00±0.10,5.34±0.57 vs 1.00±0.16;P＜0.01)when compared with the control group.Conclusion Hp infection inhibits proliferation and migra-tion,and induces apoptosis and inflammatory response in HUVEC through activation of the STAT3/NF-κB pathway.