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.

Objective:To explore the clinical characteristics and establish a corresponding prognostic scoring model in patients with early-stage clinical features of hepatitis B-induced acute-on-chronic liver failure (HBV-ACLF).Methods:Clinical characteristics of 725 cases with hepatitis B-related acute-on-chronic hepatic dysfunction (HBV-ACHD) were retrospectively analyzed using Chinese group on the study of severe hepatitis B (COSSH). The independent risk factors associated with 90-day prognosis to establish a prognostic scoring model was analyzed by multivariate Cox regression, and was validated by 500 internal and 390 external HBV-ACHD patients.Results:Among 725 cases with HBV-ACHD, 76.8% were male, 96.8% had cirrhosis base,66.5% had complications of ascites, 4.1% had coagulation failure in respect to organ failure, and 9.2% had 90-day mortality rate. Multivariate Cox regression analysis showed that TBil, WBC and ALP were the best predictors of 90-day mortality rate in HBV-ACHD patients. The established scoring model was COSS-HACHADs = 0.75 × ln(WBC) + 0.57 × ln(TBil)-0.94 × ln(ALP) +10. The area under the receiver operating characteristic curve (AUROC) of subjects was significantly higher than MELD, MELD-Na, CTP and CLIF-C ADs( P < 0.05). An analysis of 500 and 390 cases of internal random selection group and external group had similar verified results. Conclusion:HBV-ACHD patients are a group of people with decompensated cirrhosis combined with small number of organ failure, and the 90-day mortality rate is 9.2%. COSSH-ACHDs have a higher predictive effect on HBV-ACHD patients' 90-day prognosis, and thus provide evidence-based medicine for early clinical diagnosis and treatment.

Objective To evaluate the effect of argon laser photocoagulation (photocoagulation group) and endolaser photocoagulation during vitrectomy (vitrectomy group) on the eyes with Eales disease by fundus fluorescein angiography(FFA) and visual acuity (VA).Methods Sixty-seven eyes with Eales were treated by photocoagulation and 31 eyes with Eales were treated by vitrectomy. The changes of FFA and VA before and after treatment were observed.Results In the photocoagulation group, retinal non-perfusion disappeared in 31eyes(55 4%),alleviated in 17eyes(30 4%),unchanged in four eyes(7 1%)and deteriorated in four eyes(7 1%).Retinal neovascularization disappeared in 30 eyes(50 8%),alleviated in 19 eyes (32 2%),remained in six eyes(10 2%)and deteriorated in four eyes(6 8%).The first FFA after treatment, non-perfusion was existed in 25 eyes in photocoagulation group, and only was existed in two eyes in vitrectomy group.Neovascularization was existed in 29 eyes in photocoagulation group, and was existed in three eyes in vitrectomy group.The best visual correction(VA≥3 0) in photocoagulation and vitrectomy groups respectively was 30 eyes and zero eye before treatment,and after treatment respectively was 47 eyes and five eyes.There were 54 eyes and 11 eyes with VA≥0 05 in photocoagulation and vitrectomy groups respectively before treatment,and 60 eyes and 20 eyes with VA≥0 5 respectively after treatment.Conclusions The better result of FFA in vitrectomy group was related to the sufficient photocoagulation by endolaser. The higher rate of low vision in vitrectomy group was related to the impairment of macula by large amount of vitreous hemorrhages,retina detachment and vitrectomy. So the patients with Eales should be checked by FFA as early as possible.If the patients had the indicator to be treated by laser,they should be treated early and rechecked by FFA,to avoid the visual function harm by repeated vitreous hemorrhage due to incomplete treatment.