In the century-long evolution of insulin pharmaceuticals, each transformative advancement in this drug class has been closely tied to the ability to obtain new insulin isoforms for research. Despite this, the recently discovered naturally occurring isoforms of glycosylated human insulin have remained largely unattainable for proper characterization. Herein, we demonstrate for the first time that total chemical synthesis can be used to generate all isoforms. This achievement required maintaining the correct positions of the interchain disulfide bonds while effectively removing protecting groups on complex glycans. Notably, the availability of seven glycoforms reveals the important effects of natural sialylated glycans in suppressing insulin self-association and enhancing its solubility, surpassing the performance of currently employed rapid-acting insulin drugs. This work not only offers a readily adaptable platform for exploring natural O-glycosylation in other therapeutic proteins and peptides but also lays the groundwork for further research into harnessing natural glycosylation for therapeutic applications.

ObjectiveTo understand the seropositivity of neutralizing antibodies (NAb) and low-level NAb against SARS-CoV-2 infection in the community residents, and to explore the impact of COVID-19 vaccination and SARS-CoV-2 infection on the levels of NAb in human serum. MethodsOn the ground of surveillance cohort for acute infectious diseases in community populations in Shanghai, a proportional stratified sampling method was used to enroll the subjects at a 20% proportion for each age group (0‒14, 15‒24, 25‒59, and ≥60 years old). Blood samples collection and serum SARS-CoV-2 NAb concentration testing were conducted from March to April 2023. Low-level NAb were defined as below the 25^th percentile of NAb. ResultsA total of 2 230 participants were included, the positive rate of NAb was 97.58%, and the proportion of low-level NAb was 25.02% (558/2 230). Multivariate logistic regression analysis indicated that age, infection history and vaccination status were correlated with low-level NAb (all P<0.05). Individuals aged 60 years and above had the highest risk of low-level NAb. There was a statistically significant interaction between booster vaccination and one single infection (aOR=0.38, 95%CI: 0.19‒0.77). Compared to individuals without vaccination, among individuals infected with SARS-CoV-2 once, both primary immunization (aOR=0.23, 95%CI: 0.16‒0.35) and booster immunization (aOR=0.12, 95%CI: 0.08‒0.17) significantly reduced the risk of low-level NAb; among individuals without infections, only booster immunization (aOR=0.28, 95%CI: 0.14‒0.52) showed a negative correlation with the risk of low-level NAb. ConclusionsThe population aged 60 and above had the highest risk of low-level NAb. Regardless of infection history, a booster immunization could reduce the risk of low-level NAb. It is recommended that eligible individuals , especially the elderly, should get vaccinated in a timely manner to exert the protective role of NAb.

Objective:To explore the latent class of childhood abuse and recent life stress in adolescent depression patients, and its effect on intermittent theta-burst stimulation (iTBS) treatment efficacy.Methods:From August 2023 to February 2024, a total of 60 adolescent patients with depression were enrolled. The Chinese version of the maltreatment and abuse chronology of exposure scale (MACE), adolescent self-rating life events check list (ASLEC), Montgomery-Asberg depression rating scale (MADRS)and suicidal ideation scale 5 (SSI-5)were used to assess all subjects. Using latent class analysis, subjects were divided into different subgroups according to 16 factors of MACE and ASLEC. Brain network analysis was conducted using the Dosenbach brain atlas with 160 regions of interest, which were divided into seven large-scale brain subnetworks: visual network (VN), somatosensory motor network (SMN), dorsal attention network (DAN), ventral attention network (VAN), subcortical network (SCN), frontoparietal network (FPN), and default mode network (DMN). Brain network functional connectivity analysis was performed using the DPABI plugin on MATLAB R2022b platform to compare differences in functional connectivity at both edge and network levels between subjects. Data analysis was conducted using SPSS 27.0 software, employing Shapiro-Wilk test, chi-square test, independent samples t-test, and Mann-Whitney U test. Results:Based on latent class analysis, subjects were divided into high abuse-high stress group ( n=38) and low abuse-high stress group ( n=22). At baseline, the high abuse-high stress group showed significantly higher scores in SSI-5 (6.39±2.26 vs 3.45(0, 5.25)), Z=-3.628, P<0.001) and MADRS item 10 (3.00(2.00, 5.00) vs 2.00(0.75, 3.25), Z=-2.794, P=0.005) compared to the low abuse-high stress group, while there was no statistically significant difference in the total score of MADRS between the two groups (27.77±4.19, 26.59±4.74, t=1.972, P=0.053). There was no statistically significant difference in the D-value of MADRS score before and after treatment between the two groups (8.45±6.03, 7.27±5.99, t=0.729, P=0.469) and the D-value of MADRS item 10 score before and after treatment between the two groups (1.00 (0, 3.00), 1.00 (0, 1.00), Z=-1.189, P=0.234). The high abuse-high group showed significantly weakened functional connectivity at the edge level compared to low abuse-high stress group at baseline ( P<0.05, NBS corrected). In the high abuse-high stress group, the mean FC values in the differential brain regions at baseline showed significant negative correlation with baseline suicidal ideation scores ( r=-0.475, P=0.003). Conclusion:The different patterns of childhood abuse and recent life stress affect the baseline clinical presentation and brain network functional connectivity of adolescent depression patients. However, the high abuse-high stress group and low abuse-high stress group showed similar responses to iTBS treatment.

Objective: To explore the risk of endometrial polyp (EP) based on lipid metabolism and vaginal micro- ecology combined with uterine volume line drawing model. Methods: 143 EP patients treated by hysteroscopic sur- gery were selected as the experimental group , and 113 healthy women were selected as the control group at the same time. The data were randomly divided into training set and validation set according to the ratio of 7 : 3. The clinical data of the two groups were collected and recorded , and t/χ2 test , LASSO regression and multifactorial lo- gistic regression analysis were used to screen the independent risk factors , construct the prediction model , and draw the column line graph. The performance of the model was evaluated by applying subject operating characteristic (ROC) curves , calibration curves , Hosmer-Lemeshow test and clinical decision-making (DCA) curves. Results: Multifactorial logistic regression analysis showed that total cholesterol ( TC) , low-density lipoprotein cholesterol (LDL-C) , vaginal microecological balance , and uterine volume were independent risk factors for the development of EP. ROC curve analysis showed that the AUC values of the training and validation sets of the column line graph model were 0. 935 and 0. 887 , respectively , and its sensitivity and specificity were 90. 21% , 83. 46% and 86. 29% , 80. 66% respectively , The Hosmer-Lemeshow test showed that the model fits well ( training set : χ2 = 2. 261 , P = 0. 840 ; validation set : χ2 = 4. 837 , P = 0. 441) and the calibration curves of the training and validation sets were close to the ideal curves , which indicated that the model had good prediction accuracy; the analysis of DCA curves of the training and validation sets both showed that the column-line graph model had a good clinical benefit rate in predicting EP. Conclusion TC , LDL-C , vaginal microecological balance and uterine volume are independent risk factors for EP , and the column-line diagram model constructed by the model has high clinical ben- efit , calibration and accuracy in predicting the risk of EP.

Objective To explore the traditional Chinese medicine(TCM)syndrome characteristics of patients with Chikungunya hemorrhagic fever and to provide empirical data to support the application of TCM in diagnosing and treating Chikungunya hemorrhagic fever.Methods A cross-sectional survey was conducted to collect clinical data(sex,age,days since onset,and comorbidity underlying disease conditions)and TCM with four-examination information(symptoms,tongue manifestations,and pulse manifestations)from 255 patients with Chikungunya hemorrhagic fever who visited Lecong Hospital of Shunde,Foshan,the Third People's Hospital of Shunde District of Foshan,Shunde Hospital of Southern Medical University Affiliated Chencun Hospital between July 23 and July 29,2025.Factor and cluster analyses were used to summarize TCM syndrome characteristics and analyze core pathogenesis in conjunction with clinical features.Results Among the 255 patients with Chikungunya hemorrhagic fever,131 were male and 124 were female,with a age of(49.05±17.93)years and a disease duration of(3.26±1.78)days.Among the four types of examination information in TCM,35 items exhibited a frequency exceeding 10%.The most prevalent symptoms were arthralgia(180 patients,70.59%),exanthem(153 patients,60.00%),fatigue(99 patients,38.82%),anhidrosis(98 patients,38.43%),pruritus(96 patients,37.65%),and fever(92 patients,36.08%).Tongue and pulse manifestations were primarily white fur(155 patients,60.78%),pink tongue(111 patients,43.53%),slippery pulse(143 patients,56.08%),and greasy fur(134 patients,52.53%).Patients with disease onset≤3 d had a higher incidence of arthralgia,fatigue,fever,aversion to cold,generalized muscle pain,aversion to wind,insomnia,headache,sweating,low-grade fever,poor appetite,loose stool,hyperhidrosis,and red tongue than those with disease onset≥4 d(P<0.05).Patients with disease onset≥4 d had a higher incidence of pink tongue and thick fur than those with disease onset≤3 d(P<0.05).The syndrome elements in patients with Chikungunya hemorrhagic fever predominantly manifested on the defensive exterior,with involvement of the sinew-bone joints,skin-muscle,and spleen.Pathogenic factors were primarily characterized by external winds,dampness,and heat.Factor and cluster analysis result indicated three TCM pathogenesis progression patterns:imbalance of the defensive exterior with wind-dampness conflict and heat transformation;dampness-heat flowing into muscles and meridians causing joint obstruction and qi blood stasis;and dampness-heat congelation resulting in qi mechanism obstruction,consumption of body fluids,and infiltration of the skin.Conclusion Patients with Chikungunya hemorrhagic fever primarily present with fever,joint pain,and rashes.In TCM,this condition falls under the category of"dampness-warmth"syndrome.Its etiology is attributed to pathogens,with transmission occurring through mosquito bites.The core pathogenesis of TCM is the invasion of the defensive exterior and dampness-toxic heat accumulation.The therapeutic principles focus on clearing heat pathogens,resolving dampness pathogens,dispersing wind pathogens,and promoting the resolution of rashes.

BACKGROUND:Topical administration of tranexamic acid can be used for anti-skin pigmentation,but its large polarity makes it difficult to break through the cuticle barrier and cell membrane when administered topically,and the subcutaneous accumulation concentration is not easy to reach the effective therapeutic concentration.OBJECTIVE:To design functionalized self-assembled micelles to enhance the anti-pigmentation effect of tranexamic acid.METHODS:The plant polyphenol derivative epigallocatechin gallate palmitate and metformin were used as carrier materials.The self-assembled micelles with synergistic anti-pigging activity and enhanced drug permeability were prepared by hydrogen bonding and electrostatic interaction.The nanoscale properties and stability of self-assembled drug-loaded micelles were tested,and their transdermal permeability was evaluated,and their biocompatibility and cellular effects were investigated.RESULTS AND CONCLUSION:(1)Functional self-assembled drug-carrying micelles with average particle size of(176.27±5.23)nm,polydispersity coefficient of 0.23±0.02,and the Zeta potential of(-25.67±0.98)mV had good stability.The mass concentrations of tranexamic acid and metformin in the self-assembled drug-carrying micelles were(20.03±0.12)and(6.67±0.08)mg/mL,respectively.The drug loadings of tranexamic acid and metformin in the self-assembled drug-carrying micelles were(19.97±0.12)％and(6.65±0.08)％,respectively.(2)In vitro transdermal results showed that the self-assembled drug-carrying micelles had higher penetration and intradermal retention per unit skin area,and could penetrate and diffuse to deeper parts of the skin.(3)MTT assay results demonstrated that undrug-loaded self-assembled micelles containing tranexamic acid 50-250 μg/mL had no toxic effects on mouse fibroblasts and mouse skin melanoma cells.The self-assembled drug-carrying micelles containing tranexamic acid 500 μg/mL had a slight toxic effect on mouse skin melanoma cells.The self-assembled drug-carrying micelles containing 50-500 μg/mL of tranexamic acid did not cause hemolysis and had good biological safety.(4)In vitro cell culture results showed that self-assembled drug-carrying micelles containing 500 μg/mL tranexamic acid could significantly inhibit the tyrosinase activity and melanin release of mouse skin melanoma cells,and the inhibitory effect was stronger than that of tranexamic acid solution or metformin solution alone.These results indicated that functionalized self-assembled micelles could synergize with tranexamic acid to inhibit tyrosinase activity,reduce melanin synthesis,and enhance the anti-skin pigmentation effect of tranexamic acid.

Objective The quantitative analysis model of mifepristone binary mixed crystal system was established by infrared spectroscopy to improve the quality control level of Mifepristone raw material.Methods Two mifepristone polymorphs samples were prepared and characterized,and the quantitative analysis model of infrared spectral data was constructed by classical linear regression method and chemometrics method respectively.On this basis,the influence of different factors on the model quality was investigated comprehensively.Results Infrared spectroscopy combined with classical linear regression method and stoichiometric method can build a quantitative analysis model for binary mifepristone polymorphs system,and the model has good linear regression coefficient(R2)and root mean square error of cross validation(RMSECV)values.Conclusions The two polymorphs of mifepristone have independent infrared spectral characteristic peaks that can be used for quantitative study,so the classical linear regression method has more significant methodical advantages for this system.The chemometrics method is more suitable for the quantitative study of complex mixed polymorphs system.

In recent years,the demand for single enantiomer drugs has gradually increased,and the separation technology of chiral drugs has also been developed rapidly.In this article,we summarized the application and progress of cocrystal technology in chiral drug resolution,focusing on the existence forms of cocrystal former formations(single compound molecules,ions,and claths),the common preparation methods of cocrystal resolution,the physical and chemical factors affecting the resolution efficiency(co-formed chirality,solvent system,temperature)and the computer-aided prediction method of chiral cocrystal in the process of chiral drug resolution,to provide a reference for related research on chiral resolution using cocrystal technology.

Objective:To explore the latent class of childhood abuse and recent life stress in adolescent depression patients, and its effect on intermittent theta-burst stimulation (iTBS) treatment efficacy.Methods:From August 2023 to February 2024, a total of 60 adolescent patients with depression were enrolled. The Chinese version of the maltreatment and abuse chronology of exposure scale (MACE), adolescent self-rating life events check list (ASLEC), Montgomery-Asberg depression rating scale (MADRS)and suicidal ideation scale 5 (SSI-5)were used to assess all subjects. Using latent class analysis, subjects were divided into different subgroups according to 16 factors of MACE and ASLEC. Brain network analysis was conducted using the Dosenbach brain atlas with 160 regions of interest, which were divided into seven large-scale brain subnetworks: visual network (VN), somatosensory motor network (SMN), dorsal attention network (DAN), ventral attention network (VAN), subcortical network (SCN), frontoparietal network (FPN), and default mode network (DMN). Brain network functional connectivity analysis was performed using the DPABI plugin on MATLAB R2022b platform to compare differences in functional connectivity at both edge and network levels between subjects. Data analysis was conducted using SPSS 27.0 software, employing Shapiro-Wilk test, chi-square test, independent samples t-test, and Mann-Whitney U test. Results:Based on latent class analysis, subjects were divided into high abuse-high stress group ( n=38) and low abuse-high stress group ( n=22). At baseline, the high abuse-high stress group showed significantly higher scores in SSI-5 (6.39±2.26 vs 3.45(0, 5.25)), Z=-3.628, P<0.001) and MADRS item 10 (3.00(2.00, 5.00) vs 2.00(0.75, 3.25), Z=-2.794, P=0.005) compared to the low abuse-high stress group, while there was no statistically significant difference in the total score of MADRS between the two groups (27.77±4.19, 26.59±4.74, t=1.972, P=0.053). There was no statistically significant difference in the D-value of MADRS score before and after treatment between the two groups (8.45±6.03, 7.27±5.99, t=0.729, P=0.469) and the D-value of MADRS item 10 score before and after treatment between the two groups (1.00 (0, 3.00), 1.00 (0, 1.00), Z=-1.189, P=0.234). The high abuse-high group showed significantly weakened functional connectivity at the edge level compared to low abuse-high stress group at baseline ( P<0.05, NBS corrected). In the high abuse-high stress group, the mean FC values in the differential brain regions at baseline showed significant negative correlation with baseline suicidal ideation scores ( r=-0.475, P=0.003). Conclusion:The different patterns of childhood abuse and recent life stress affect the baseline clinical presentation and brain network functional connectivity of adolescent depression patients. However, the high abuse-high stress group and low abuse-high stress group showed similar responses to iTBS treatment.

Objective To explore the traditional Chinese medicine(TCM)syndrome characteristics of patients with Chikungunya hemorrhagic fever and to provide empirical data to support the application of TCM in diagnosing and treating Chikungunya hemorrhagic fever.Methods A cross-sectional survey was conducted to collect clinical data(sex,age,days since onset,and comorbidity underlying disease conditions)and TCM with four-examination information(symptoms,tongue manifestations,and pulse manifestations)from 255 patients with Chikungunya hemorrhagic fever who visited Lecong Hospital of Shunde,Foshan,the Third People's Hospital of Shunde District of Foshan,Shunde Hospital of Southern Medical University Affiliated Chencun Hospital between July 23 and July 29,2025.Factor and cluster analyses were used to summarize TCM syndrome characteristics and analyze core pathogenesis in conjunction with clinical features.Results Among the 255 patients with Chikungunya hemorrhagic fever,131 were male and 124 were female,with a age of(49.05±17.93)years and a disease duration of(3.26±1.78)days.Among the four types of examination information in TCM,35 items exhibited a frequency exceeding 10%.The most prevalent symptoms were arthralgia(180 patients,70.59%),exanthem(153 patients,60.00%),fatigue(99 patients,38.82%),anhidrosis(98 patients,38.43%),pruritus(96 patients,37.65%),and fever(92 patients,36.08%).Tongue and pulse manifestations were primarily white fur(155 patients,60.78%),pink tongue(111 patients,43.53%),slippery pulse(143 patients,56.08%),and greasy fur(134 patients,52.53%).Patients with disease onset≤3 d had a higher incidence of arthralgia,fatigue,fever,aversion to cold,generalized muscle pain,aversion to wind,insomnia,headache,sweating,low-grade fever,poor appetite,loose stool,hyperhidrosis,and red tongue than those with disease onset≥4 d(P<0.05).Patients with disease onset≥4 d had a higher incidence of pink tongue and thick fur than those with disease onset≤3 d(P<0.05).The syndrome elements in patients with Chikungunya hemorrhagic fever predominantly manifested on the defensive exterior,with involvement of the sinew-bone joints,skin-muscle,and spleen.Pathogenic factors were primarily characterized by external winds,dampness,and heat.Factor and cluster analysis result indicated three TCM pathogenesis progression patterns:imbalance of the defensive exterior with wind-dampness conflict and heat transformation;dampness-heat flowing into muscles and meridians causing joint obstruction and qi blood stasis;and dampness-heat congelation resulting in qi mechanism obstruction,consumption of body fluids,and infiltration of the skin.Conclusion Patients with Chikungunya hemorrhagic fever primarily present with fever,joint pain,and rashes.In TCM,this condition falls under the category of"dampness-warmth"syndrome.Its etiology is attributed to pathogens,with transmission occurring through mosquito bites.The core pathogenesis of TCM is the invasion of the defensive exterior and dampness-toxic heat accumulation.The therapeutic principles focus on clearing heat pathogens,resolving dampness pathogens,dispersing wind pathogens,and promoting the resolution of rashes.