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.

Hypoxic-ischemic (HI) brain damage poses a high risk of death or lifelong disability, yet effective treatments remain elusive. Here, we demonstrated that miR-100-5p levels in the lesioned cortex increased after HI insult in neonatal mice. Knockdown of miR-100-5p expression in the brain attenuated brain injury and promoted functional recovery, through inhibiting the cleaved-caspase-3 level, microglia activation, and the release of proinflammation cytokines following HI injury. Engineered extracellular vesicles (EVs) containing neuron-targeting rabies virus glycoprotein (RVG) and miR-100-5p antagonists (RVG-EVs-Antagomir) selectively targeted brain lesions and reduced miR-100-5p levels after intranasal delivery. Both pre- and post-HI administration showed therapeutic benefits. Mechanistically, we identified protein phosphatase 3 catalytic subunit alpha (Ppp3ca) as a novel candidate target gene of miR-100-5p, inhibiting c-Fos expression and neuronal apoptosis following HI insult. In conclusion, our non-invasive method using engineered EVs to deliver miR-100-5p antagomirs to the brain significantly improves functional recovery after HI injury by targeting Ppp3ca to suppress neuronal apoptosis.
Animals ; MicroRNAs/metabolism* ; Extracellular Vesicles/metabolism* ; Mice ; Recovery of Function/physiology* ; Hypoxia-Ischemia, Brain/therapy* ; Mice, Inbred C57BL ; Antagomirs/administration & dosage* ; Male ; Animals, Newborn ; Apoptosis/drug effects* ; Brain Injuries/metabolism* ; Glycoproteins ; Peptide Fragments ; Viral Proteins

Objective:To explore the impact of air pollution on hospital visits for mental and behavioral disorders among residents in an industrial area in Henan Province from 2016 to 2021.Methods:Daily outpatient visits data for mental and behavioral disorders were collected from Angang General Hospital in Angang Industrial Area at Anyang City between January 2016 and December 2021. And air pollutants and meteorological data during the same period were also collected. A generalized additive model was used for time-series analysis to examine the relationship between daily average concentrations of nitrogen dioxide (NO 2), sulfur dioxide (SO 2), fine particulate matter (PM 2.5), inhalable particulate matter (PM 10), carbon monoxide (CO), and ozone (O 3) with a lag of 0 to 7 days on the number of visits for mental and behavioral disorders among residents. The single-day lag effect (lag0-lag7 d) and cumulative lag effect (lag01-lag07 d) were analyzed. The smooth cubic spline function was used to fit the exposure-response relationship, and subgroup analysis was performed according to different genders, seasons and ages. Results:A total of 26 268 hospital visits for mental and behavioral disorders were collected from the industrial area between 2016 and 2021. The daily average concentrations of SO 2, NO 2, PM 2.5, PM 10, and CO were (27.50±27.33), (43.11±18.33), (73.87±60.30), (134.01±83.81) μg/m 3, and (1.72±1.03) mg/m 3, respectively. The daily maximum 8-hour average concentration of O 3 was (82.18±53.70) μg/m 3. After controlling for long-term trends, temperature, relative humidity, day of the week effects, and holiday effects, the generalized additive model analysis showed that NO 2 had a statistically significant impact on the hospital visits for mental and behavioral disorders at lag0 d, lag2 d and lag01-lag05 d and CO had a statistically significant impact at lag0-lag3 d and lag01-lag06 d (all P<0.05). NO 2 at lag02-lag04 d and CO at lag0-lag2 d and lag01-lag04 d had statistically significant effects on the visits for neurasthenia (both P<0.05). The impacts of NO 2 at lag03-lag04 d, PM 2.5 at lag3 d and lag03-lag04 d, PM 10 at lag3 d and lag03 d, and CO at lag3 d and lag01-lag05 d on visits for generalized anxiety disorder were also statistically significant (all P<0.05). After false discovery rate (FDR) correction, it was shown that for every 10 μg/m 3 increase in NO 2 and every 0.1 mg/m 3 increase in CO, the percentage increase in visits for mental and behavioral disorders and its 95% confidence interval (95% CI) were 3.38% (0.95%-5.87%) and 0.78% (0.38%-1.17%), respectively. For every 0.1 mg/m 3 increase in CO, the visits for neurasthenia increased by 0.78% (0.27%-1.29%). For every 10 μg/m 3 increase in PM 2.5 and every 0.1 mg/m 3 increase in CO, the visits for generalized anxiety disorder increased by 1.07% (0.46%-1.68%) and 1.17% (0.37%-1.97%), respectively (adjusted P<0.05). There was a linear exposure-response relationship between NO 2 and CO and the hospital visits for mental and behavioral disorders, CO and the hospital visits for neurasthenia, and CO and PM 2.5 and the hospital visits for generalized anxiety disorder ( P<0.05 for the overall association test and P>0.05 for the non-linearity test). Stratified analysis showed that air pollutants had an impact on male patients with neurasthenia, female patients with generalized anxiety disorder, individuals aged <45 years with mental and behavioral disorders, and individuals aged ≥65 years with generalized anxiety disorder. The impact of air pollutants was greater during the cold season or winter. Conclusion:Exposure to air pollution can increase hospital visits for mental and behavioral disorders among residents in industrial areas, with a higher risk among those aged<45 years old and during the cold season.

BACKGROUND:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

Objective To explore the value of CT signs and quantitative parameters of artificial intelligence (AI) in predicting the efficacy of neoadjuvant chemotherapy for colorectal cancer. Methods A total of 349 colorectal cancer patients who received neoadjuvant chemotherapy at Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine in Hebei Province from January 2022 to January 2025 were selected and and divided into the effective group (n = 267) and the ineffective group (n = 82) according to the evaluation criteria for the efficacy of solid tumors. Conduct a CT examination and extract AI quantitative parameters from the CT images based on the lesion. The data were analyzed using SPSS21.0 software, Logistic regression was used to screen the influencing factors of ineffective neoadjuvant chemotherapy in patients with colorectal cancer, and separate and combined models of CT signs and AI quantitative parameters were established. The predictive effect of the model was verified by using the ROC curve, calibration curve and decision curve. Results Compared with the effective group, the proportion of regular tumor morphology and the proportion of non-enlarged lymph nodesin the ineffective group were smaller. The tumor volume, peak value and entropy value were larger (P < 0.05). Multivariable analysis showed that irregular shape (OR= 4.216), presence of lymph node enlargement (OR = 8.998), larger tumor volume (OR = 1.109), higher average CT value (OR = 1.120), elevated peak value (OR = 2.528), and increased entropy value (OR = 1.390) were independent risk factors for ineffective neoadjuvant chemotherapy in colorectal cancer (P < 0.05). The areas under the ROC curves of the individual and combined models of CT signs and AI quantitative parameters were 0.777, 0.818, and 0.877, respectively(P < 0.05). The calibration curve showed a Brier score of 0.091. The decision curve showed that the threshold was between 0.10 and 0.85, and the combined model achieved a relatively high net clinical benefit. Conclusion CT signs combined with AI quantitative parameters has a predictive value for the efficacy of neoadjuvant chemotherapy in colorectal cancer. To provide evidence-based basis for clinical screening of the population benefiting from chemotherapy and optimization of treatment strategies.

Objective:To investigate the predictive value of different obesity indicators for colorectal cancer (CRC) risk in different gender populations.Methods:This observational study was conducted within the Kailuan Study (Registration Number: ChiCTR-TNC-11001489). From July 2006 to October 2007, a total of 101,510 employed and retired individuals underwent health examinations, including gastrointestinal disease screening, hematological tests, and questionnaires, at Kailuan General Hospital and its 10 affiliated hospitals. After excluding those with incomplete data, 93,606 participants were included in this study and divided into male (74 852) and female (18 754) groups. CRC incidence was collected through physical examinations and questionnaires every two years. Each participant's follow-up period began at the time of the questionnaire and ended upon CRC diagnosis, death, or December 31, 2021. Body Mass Index (BMI), waist circumference, waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR) were quartiled (Q1, Q2, Q3, Q4), with Q1 serving as the control group. After adjusting for traditional risk factors such as age, total cholesterol, triglycerides, diabetes, hypertension, smoking status, alcohol consumption, and physical exercise, Cox regression models were used to calculate the correlations between BMI, waist circumference, WHR, WHtR, and CRC incidence in both male and female populations.Results:The age of all patients was (51±12) years, BMI was (25.06±3.49) kg/m 2, waist circumference was (86.94±9.97) cm, hip circumference was (97.30±8.81) cm, WHR was 0.89±0.07, and WHtR was 0.52±0.06.Female participants had significantly lower BMI, waist circumference, WHR, and WHtR compared to males, with statistically significant differences (all P<0.05). The mean follow-up duration for all participants was 15.01 (14.10±2.66) years, during which 718 CRC cases were identified, including 626 males (0.83%) and 92 females (0.49%). Cox proportional hazards models for males showed that CRC risk increased with waist circumference from Q3 (HR=1.43, 95%CI: 1.13-1.79, P=0.003) to Q4 (HR=1.45,95%CI: 1.14-1.82, P=0.002). Similarly, CRC risk increased with WHR from Q3 (HR=1.22, 95%CI: 1.01-1.53, P=0.007) to Q4 (HR=1.43, 95%CI: 1.14-1.79, P=0.002) and with WHtR from Q3 (HR=1.37, 95%CI: 1.08-1.74, P=0.009) to Q4 (HR=1.68, 95%CI: 1.33-2.12, P<0.001). For females, CRC risk increased with waist circumference from Q2 (HR=2.37, 95%CI: 1.20-4.67, P=0.012) to Q3 (HR=2.42, 95%CI: 1.21-4.84, P=0.013) but decreased in Q4 ( HR=2.08, 95%CI: 1.02-4.25, P=0.043). CRC risk increased significantly with WHR from Q2 (HR=2.20, 95%CI: 1.11-4.39, P=0.024) to Q3 (HR=2.89, 95%CI: 1.48-5.67, P=0.002) in females but was not statistically significant in Q4 ( P=0.074). Among females, CRC risk also increased significantly with WHtR in Q2 (HR=2.30, 95% CI: 1.16-4.56, P=0.017) and Q4 (HR=2.64, 95%CI: 1.32-5.29, P=0.006). There were no statistically significant differences in CRC risk associated with BMI in either male or female populations (both P>0.05). Conclusion:Waist circumference, WHR, and WHtR were better predictors of CRC risk than BMI in both male and female populations.

Objective:To explore the correlation between metabolic syndrome (MS), serum high-sensitivity C-reactive protein (hs-CRP) levels, their combination and the risk of digestive system malignancies.Methods:A prospective cohort study was conducted in the participants from the Kailuan cohort who took health examination in July 2006. Anthropometric parameters, epidemiological information, and laboratory test results were collected. Incidence and mortality of digestive system malignant tumors were collected through biennial health examinations and questionnaires. The follow-up period ended on December 31, 2021.According to MS status and hs-CRP levels (hs-CRP≤3 or >3 mg/L), the cohort was divided into 4 groups, induding MS -hs-CRP -, MS -hs-CRP +, MS + hs-CRP -, and MS + hs-CRP + group. Chi-squared test, one analysis of variance, and the Kruskal-Wallis H test were used for inter-group comparison among groups. Kaplan-Meier method was used to calculate the cumulative incidence of digestive system malignant tumors, and log-rank test was performed to compare the cumulative incidence among groups. Multivariable Cox proportional hazards regression models were used to evaluate the effects of MS and hs-CRP levels on the overall risk of digestive system malignant tumors, as well as the effects of their combination on the risk of digestive system malignant tumors of different site, and relevant confounding factors were adjusted.A sensitivity analysis was conducted by excluding individuals diagnosed with digestive system malignancies within one year of follow-up, as well as those taking antihypertensive, antidiabetic, or lipid-lowering medications. Results:A total of 92 916 participants were included in this study. Among them, 57 933 cases were in the MS -hs-CRP - group, 10 949 cases in the MS -hs-CRP + group, 18 412 cases in the MS + hs-CRP - group, and 5 622 cases in the MS + hs-CRP + group.The median follow-up period was 15.01 years (14.66 to 15.20 years). By the end of follow-up, these were 1 992 cases of new-onset digestive system malignant tumors. The cumulative incidence rates of digestive system malignant tumors of MS -hs-CRP -, MS -hs-CRP +, MS + hs-CRP -, and MS + hs-CRP + groups were 2.0%(1 164/57 933), 2.3%(249/10 949), 2.4%(440/18 412), and 2.5%(139/5 622), respectively. The difference in the cumulative incidence among the 4 groups was statistically significant ( χ2=14.09, P=0.003).The results of multivariate Cox analysis showed that, after hs-CRP level and other confounding factors were adjusted, the risk of developing digestive system malignant tumors in participants with MS was 21.4% higher than that in those without MS ( HR=1.214 (95% confidence interval (95% CI): 1.086 to 1.340), P<0.001). After MS status and other confounding factors were adjusted, the risk of developing digestive system malignant tumors in participants with high hs-CRP level (>3 mg/L) was 17.2% higher than those with low hs-CRP level (≤3 mg/L) ( HR=1.172 (95% CI: 1.042 to 1.303), P=0.008). After relevant confounding factors were adjusted, the risks of developing digestive system malignant tumors in the MS -hs-CRP +, MS + hs-CRP -, and MS + hs-CRP + groups increased by 17.2%, 21.4%, and 35.9%, respectively, as compared with that of the MS -hs-CRP - group ( HR=1.172 (95% CI: 1.017 to 1.399), P=0.028; HR=1.214 (95% CI: 1.074 to 1.356), P=0.002; HR=1.359 (95% CI: 1.135 to 1.635), P=0.001). Among the 4 groups, the overall risk of developing digestive system malignant tumors of MS + hs-CRP + group was the highest. After relevant confounding factors were adjusted, the risks of colorectal cancer, liver cancer, and pancreatic cancer of the MS + hs-CRP + group increased by 46.2%, 35.7%, and 88.3%, respectively, as compared with those of the MS -hs-CRP - group ( HR=1.462 (95% CI: 1.088 to 1.956), HR=1.357 (95% CI: 1.132 to 2.089), HR=1.883 (95% CI: 1.052 to 3.342)), suggesting that MS combined with high hs-CRP was a significant risk factor for increased incidences of colorectal cancer, liver cancer, and pancreatic cancer ( P=0.012, 0.016 and 0.033). After participants diagnosed with new digestive system malignancies within one year of follow-up and those taking antihypertensive, antidiabetic, or lipid-lowering medications (108 cases, 10 680 cases, 2 344 cases, 906 cases) were excluded, the results of sensitivity analysis indicated the increased risk of digestive system malignant tumors in the MS -hs-CRP +, MS + hs-CRP -, and MS + hs-CRP + groups were 12.1%, 21.4%, 28.7%; 18.2%, 21.4%, 24.8%; 16.4%, 21.4%, 32.2%; 17.3%, 20.4%, 35.8%. Among the 3 groups, the increased risk of developing digestive system malignant tumors of MS + hs-CRP + group was the highest. Conclusion:MS and hs-CRP >3 mg/L are both independent risk factors for developing digestive system malignant tumors, and their combination further increases the risk of developing digestive system malignant tumors.

Objective:To analyze the epidemiological characteristics and survival rate of nasopharynx cancer (NPC) in Guangdong Province from 2011 to 2019.Methods:Based on the cancer registry data of Guangdong Province from 2011 to 2019, the crude rate, age-standardized rate (the standard population was the fifth Chinese national census of 2000) and age-specific rate of incidence and mortality of NPC were calculated, and the regional distribution characteristics were also explored. The average annual percentage change (AAPC) of the incidence and mortality rates were analyzed by using Joinpoint regression model. The observed survival rate was estimated by period survival method, and the expected survival rate was calculated by Ederer Ⅱ method.Results:The crude incidence rate and age standardized incidence rate of NPC showed a decreasing trend, and the AAPC was -1.9% and -2.1%, respectively ( P＜0.05). The crude mortality rate and age standardized mortality rate of NPC also showed a decreasing trend, and the AAPC was -4.8% and -4.6%, respectively ( P＜0.05). The incidence and mortality rates are both higher in men than those in women during the nine years. The age-specific incidence rate of NPC reached its peak in the 50-64 years old age group, and the mortality rate reached its peak in the 65-74 years old age group in Guangdong province. In 2019, the age-standardized incidence rate of NPC was 9.49/100 000 (13.89/100 000 in men and 5.19/100 000 in women). The incidence and mortality of NPC varied greatly among different areas, and the areas with highest incidence and mortality rate were both in Zhaoqing. In 2020, the five-year observed survival rate of NPC in Guangdong Province was 67.2%, the 5-year relative survival rate was 75.3% and the 5-year standardized relative survival rate was 68.9%. Conclusions:Both the incidence and mortality rates of NPC in Guangdong province show decreasing trend, and the decreasing level of the mortality rate is higher than that of the incidence rate, but the two rates are still at high levels. The prevention and control work should focus on male, middle-aged and elderly population and Zhaoqing, Zhongshan, Foshan areas.

Objective To analyze the effects of fluid overload(FO)on clinical outcomes of extremely preterm infants with hemody-namically significant patent ductus arteriosus(hsPDA).Methods This retrospective study analyzed data from neonates ≤ 32 weeks ges-tational age(GA)admitted to the Neonatal Intensive Care Unit(NICU)of the Affiliated Hospital of Xuzhou Medical University between November 2021 and July 2024.Infants were categorized into hsPDA and non-hsPDA groups based on the presence or absence of hsPDA.General clinical data and daily FO situation in the first week after birth of the two groups were compared,and the risk factors affecting hsPDA were obtained by Logistic regression analysis.Using the first day of postnatal FO as the test variable and hsPDA as the state varia-ble,receiver operator characteristic(ROC)curve was drawn to calculate the cut-off value of FO,and the cut-off value was divided into groups to analyze the relationship between the first day FO and clinical outcome of very preterm infants.Results A total of 153 extremely preterm infants(GA≤32 weeks)were enrolled,including 110 in the hsPDA group and 43 in the non-HSPDA group.Univariate analysis showed that there were statistically significant differences in birth weight,1-minute Apgar score,postnatal antibiotic use time,parenteral nutrition time,invasive mechanical ventilation time,non-invasive mechanical ventilation time,hospital days,day 1,day 2 and day 4 FO between the two groups(P＜0.05).Multivariate Logistic regression analysis showed that invasive mechanical ventilation time and day 1 FO were risk factors of hsPDA.The ROC curve was drawn with the first day FO as the test variable and hsPDA as the state variable.The area under the curve was 0.903,the sensitivity and specificity were 79.1％and 89.1％,respectively,and the cut-off value of FO on the first day after birth was 8.78％.According to this cut-off value,107 cases were divided into FO≤8.78％group and 46 cases were FO＞8.78％group.Univariate analysis showed that FO＞8.78％group had lower Apgar score at 1 minute,longer invasive mechani-cal ventilation time,and higher incidence of moderate to severe bronchopulmonary dysplasia(BPD)and grade 3 to 4 intraventricular hem-orrhage(IVH).Logistic regression analysis showed that FO＞8.78％on the first day after birth was a risk factor for moderate to severe BPD and grade 3-4 IVH.Conclusion FO＞8.78％on the first day after birth of very preterm infants increased the incidence of moder-ate to severe BPD and grade 3-4 IVH.

Objective To quantitatively evaluate the structure and content of the long-term care insurance(LTCI)policy in China's deeply aging areas.Methods Using the Policy Modeling Consistency(PMC)index model indicator design method,a LTCI policy evalua-tion index system was constructed,consisting of nine primary indicators and 34 secondary indicators.A total of 123 provincial-level LTCI policies issued in deeply aging regions of China between June 1,2014,and October 1,2024 were analyzed.High-frequency word extraction was performed using ROSTCM 6.0,and a social network diagram of LTCI policies was created.The policy structure and content were quantitatively evaluated and ana-lyzed based on the established policy evaluation index system.Results The main content of LTCI policies in deeply aging areas of China covered services,institutions and assessment.The highest policy score was 7.28,and the lowest was 2.20,with an average score of 5.00.There were 25 perfect policies,63 excellent policies,28 good policies and seven qualified policies.In the dimension of policy content,the indexes of five primary indicators of policy evaluation,policy target groups,policy nature,policy perspective and policy tools were 0.60 or more;while the indexes of four primary indicators of policy content,incentives and constraints,policy timeliness,and policy level were 0.50 or less.Conclusion LTCI policies issued in China's deeply aging areas provide comprehensive coverage in aspects such as poli-cy evaluation,policy target groups and policy nature,and need to be improved in policy tool selection and the construction of incentive and constraint mechanisms.