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 retrieve, evaluate and summarize the best evidence on the use of bedside ultrasound by ICU nurses to assess the lungs of adult critically ill patients, and to provide a reference for clinical practice and the construction of related processes and protocols.Methods:Based on the "6S" pyramid model, a computer-based search was conducted on relevant computer decision support system, guideline networks, professional associations, and domestic and international databases, the search time limit was from the establishment of the database to June 5, 2024. The panel members who had been trained in the evidence-based course evaluated the included literature with corresponding tools, extracted evidence according to the theme.Results:Twenty-five papers were finally included, including 6 guidelines, 8 expert consensus, 2 expert opinion, 3 clinical decision-making, 3 systematic evaluation, and 3 randomized controlled trials. A total of 35 pieces of evidence were formed from 4 aspects, including personnel training, operation specifications, clinical application (including dyspnea screening, intervention implementation, efficacy evaluation, diaphragm function evaluation) and precautions.Conclusions:The best evidence for lung assessment and intervention in adult critically ill patients based on bedside ultrasound can provide a reference for the adjustment and decision-making of nursing measures for adult critically ill patients. In the subsequent process of evidence transformation, attention should be paid to combining clinical practice and the joint cooperation of medical staff.

Objective:To retrieve, evaluate and summarize the best evidence on the use of bedside ultrasound by ICU nurses to assess the lungs of adult critically ill patients, and to provide a reference for clinical practice and the construction of related processes and protocols.Methods:Based on the "6S" pyramid model, a computer-based search was conducted on relevant computer decision support system, guideline networks, professional associations, and domestic and international databases, the search time limit was from the establishment of the database to June 5, 2024. The panel members who had been trained in the evidence-based course evaluated the included literature with corresponding tools, extracted evidence according to the theme.Results:Twenty-five papers were finally included, including 6 guidelines, 8 expert consensus, 2 expert opinion, 3 clinical decision-making, 3 systematic evaluation, and 3 randomized controlled trials. A total of 35 pieces of evidence were formed from 4 aspects, including personnel training, operation specifications, clinical application (including dyspnea screening, intervention implementation, efficacy evaluation, diaphragm function evaluation) and precautions.Conclusions:The best evidence for lung assessment and intervention in adult critically ill patients based on bedside ultrasound can provide a reference for the adjustment and decision-making of nursing measures for adult critically ill patients. In the subsequent process of evidence transformation, attention should be paid to combining clinical practice and the joint cooperation of medical staff.

Objective:To translate the University of Rhode Island Change Assessment of voice scale（URICA-Voice） into Chinese and test its reliability and validity. Methods:The URICA-Voice scale was converted into Chinese by literal translation, cultural adjustment, expert consultation, pre-investigation, and back translation. Convenience sampling was used to recruit patients at four speech therapy centers from February to May 2022. Then the Chinese version of the scale was distributed to them, and the reliability and validity of the scale were tested after data collection. Cronbach ɑ was used to evaluate the reliability. The critical ratio method and Pearson correlation coefficient were used for item analysis. Item-level content validity, scale-level content validity, and confirmatory factor analysis were used to test the validity of the scale. Results:A total of 247 valid questionnaires were collected. ①Item analysis: the critical ratios between a high-score and low-score groups of 32 items were all statistically significant（P<0.01） and all the critical ratios were above 3.00. The Pearson correlation between 32 items and the total score was significant（P<0.01）. ②Validity analysis: I-CVI=1.00, S-CVI/Ave=1.00, χ²/df=2.30, RMSEA=0.07. Except for item 9 and 23, the standardized factor loading coefficients of other items were all above 0.50. AVE of the four dimensions of the scale was all above 0.50, and the combined reliability of the four dimensions was all above 0.70. The correlation coefficients between dimensions were less than the square root of the AVE of the dimension itself. ③Reliability analysis: the Cronbach ɑ of the whole scale was 0.94, and the Cronbach ɑ of the four dimensions were 0.88, 0.92, 0.94, and 0.88 respectively. Conclusion:The Chinese version of URICA-Voice has good reliability and validity, and can be used as a specific measurement tool for evaluating the compliance of voice training in China.
Humans ; China ; Language ; Reproducibility of Results ; Surveys and Questionnaires ; Voice

Objective: To investigate the effects of calcium supplementation during the pregnancy and early infancy stage on body mass index (BMI) and gut microbiota in the infants. Methods: A total of 1 752 healthy pregnant women and their infants (breast feeding) in two maternal and child health care hospitals of Beijing were chosen as the subjects in this study from May to October 2016. Questionnaires were used to obtain the general information and supplementation of calcium and vitamin D in mothers and their infants. The body length and weight of infants at birth and 6 months were recorded to calculate the BMI. The random number table method was used to randomly select 40 infants from each group for gut microbiota analysis (If less than 40 infants were all included in this study, 23 infants in the pregnancy and early infancy would be all treated with calcium supplements. There were 6 infants who was not added calcium during the pregnancy but added in the early infancy). Then it was compared that the effects of calcium supplementation during the pregnancy and early infancy on the BMI and gut microbiota composition of infants were determined at birth and 6 months. Results: Compared to the group with no calcium supplementation during the pregnancy ((12.76±1.23), (17.68±0.76)kg/m²), the BMI of infants at birth and 6 months in the group with calcium supplementation during the pregnancy ((13.51±0.47), (17.91±0.23)kg/m²) were significantly higher(P<0.05). In the group with maternal calcium supplementation, the BMI at 6 months ((18.63±0.52)kg/m²), BMI increment ((5.71±0.54)kg/m²) and the content of lactobacillus (21.04%±3.68%) in the only calcium supplementation subgroup in the early infancy were higher than those in only vitamin D supplementation subgroup ((17.69±0.89) kg/m², (4.17±1.01) kg/m² and 12.28%±3.86%) (P<0.05). In the group without maternal calcium supplementation, the content of lactobacillus (20.15%±4.87%) in the only calcium supplementation subgroup were also higher than those in only vitamin D supplementation subgroup (14.64%±3.71%) (P<0.05). Conclusion Appropriate calcium supplementation during the pregnancy is good for the growth and development of the fetus. Calcium supplementation in the early infancy could increase the BMI of infants, and promote the growth of intestinal lactobacillus.

The penetrating moxibustion technique is proposed based on experience of the ancients and clinical practice for many years. From the aspects of definition, action characteristics and technique at different parts, the advantage and application experience of penetrating moxibustion have been discussed. The adequate dose of moxibustion is necessary in penetrating moxibustion; in addition, moxibustion sensation should be penetrating; therefore, with a temperature of 43℃ for more than 20 min, sweating, flushing, speckle appearing after penetrating moxibustion. Due to individual differences of age, gender and constitution factors, the effects of penetrating moxibustion are different, in clinical practice the body reaction and patient's feelings should be deliberately focused on other than does or sore and blister of moxibustion. The technique is common used in the abdomen, waist and knee joint, etc.

CRISPR-Cas Systems ; Embryo, Mammalian ; Gene Editing ; methods ; HEK293 Cells ; Humans ; Polycomb Repressive Complex 1 ; genetics ; Zygote

OBJECTIVETo observe the clinical efficacy differences between acupuncture combined with 40-min penetration moxibustion and 60-min penetration moxibustion at back-points for insomnia of insufficiency of heart and spleen type.

METHODSSixty patients of insomnia with insufficiency of heart and spleen type were randomly assigned into a 40-min group and a 60-min group. The two groups were treated with acupuncture at Jueyinshu (BL 14), Xinshu (BL 15), Geshu (BL 17), Pishu (BL 20), Shendao (GV 11) and Zhiyang (GV 9). With moxibustion box, the penetration moxibustion was applied at the back until sweating and redness on the back. The moxibustion was given for 40 min in the 40-min group and 60 min in the 60-min group. The treatment was given once a day, five days per week. Each session was consisted of 5 treatments, with an interval of 2 days between session and totally 4 consecutive weeks were provided. The Pittsburgh sleep quality index (PSQI), TCM symptom scale were observed and recorded before and after treatment in the two groups. The even temperature at raising period, effective period, reducing period, as well as minimum high temperature, comfortable temperature, minimum cold temperature and medication status were compared; also the effect was compared between the two groups.

RESULTSThe total effective rate was 96.6% (28/29) in the 60-min group, which was higher than 89.3% (25/28) in the 40-min group (<0.05). Compared before treatment, the total score of PSQI and sleep quality, sleep time, sleep efficiency, sleep disorder, daytime dysfunction as well as the total TCM symptom score and its drowsiress in the morning, palpitation, amnesia, appetite were reduced after treatment in the 40-min group (all<0.05). After treatment, the total score and each score of PSQI as well as total score and each score of TCM symptom scale were reduced after treatment in the 60-min group (all<0.05). After treatment, the total score and each score of PSQI as well as total score and each score of TCM symptom scale were significantly different between the two groups (all<0.05).

CONCLUSIONSAcupuncture combined with penetration moxibustion can improve the symptomsof insomnia with insufficiency of heart and spleen type, which is more significant in the 60-min group, indicating prolonged time of penetration moxibustion can improve sleep latency.

Objective To investigate the effects of nimotuzumab on radiosensitivity of ECA?109 and TE?13 esophageal carcinoma cell lines and explore its possible mechanism. Methods The ECA?109 and TE?13 cells were divided into control group, irradiation group, medicine group, and combined group ( irradiation + medicine) . In the combined group, ECA?109 and TE?13 cells were treated with nimotuzumab for 24 h before irradiation, and the cells were collected 2 h after irradiation. The radiosensitizing effects of nimotuzumab on ECA?109 and TE?13 cells were evaluated by clone formation assay. Cell apoptosis was detected by flow cytometry. Western blotting was used to evaluate the expression of EGFR, p?EGFR, DNA?PKcs, p?DNA?PKcs and γH2AX. Results The values of Dq( quasithreshold dose) , D0( mean lethal dose) and SF2(surviving fraction at 2 Gy) of ECA?109 and TE?13 cells in the combined group were significantly lower than those of the radiation group ( for ECA?109 cells, 1. 11 vs. 1. 72, 1. 40 vs. 2. 14, 0. 42 vs. 0. 66, respectively;for TE?13 cells, 0.41 vs. 0.46, 0.43 vs. 0.65, 0.40 vs. 0.71, respectively ( all P<0.05) . The sensitivity enhancement ratio (SER) of ECA?109 and TE?13 cells were 1.35 and 1.43, respectively. Flow cytometry showed that the apoptosis rate of ECA?109 and TE?13 cells in the combined group were significantly higher than those of the radiation group [ for ECA?109 cells, ( 41. 31 ± 1. 52)% vs. ( 9. 54 ± 0.52)%;for TE?13 cells, (46.28±0.28)% vs. (11.32±0.31)%, both P<0.01]. Western blotting showed that the expression levels of EGFR and DNA?PKcs were not significantly different in all groups ( all P>0.05) . Compared with those of the control group, p?EGFR and p?DNA?PKcs of the radiation group were significantly higher in both cell lines ( P<0.05) , and theγH2AX levels in the radiation group and medicine group were significantly higher than that of the control group ( P<0.05) . Compared with those of the radiation group and medicine group, p?EGFR and p?DNA?PKcs protein expression in the combined group were decreased significantly (P<0.05), while γH2AX protein expression was significantly increased (P<0.05). Conclusions Nimotuzumab can enhance the radiosensitivity of esophageal cancer ECA?109 and TE?13 cells. The potential mechanism may be related to the inhibition of EGFR phosphorylation and down?regulation of DNA damage repair proteins. The radiosensitizing effect of nimotuzumab is greater on poorly differentiated esophageal cancer cells.

Objective To investigate the effects of nimotuzumab on radiosensitivity of ECA?109 and TE?13 esophageal carcinoma cell lines and explore its possible mechanism. Methods The ECA?109 and TE?13 cells were divided into control group, irradiation group, medicine group, and combined group ( irradiation + medicine) . In the combined group, ECA?109 and TE?13 cells were treated with nimotuzumab for 24 h before irradiation, and the cells were collected 2 h after irradiation. The radiosensitizing effects of nimotuzumab on ECA?109 and TE?13 cells were evaluated by clone formation assay. Cell apoptosis was detected by flow cytometry. Western blotting was used to evaluate the expression of EGFR, p?EGFR, DNA?PKcs, p?DNA?PKcs and γH2AX. Results The values of Dq( quasithreshold dose) , D0( mean lethal dose) and SF2(surviving fraction at 2 Gy) of ECA?109 and TE?13 cells in the combined group were significantly lower than those of the radiation group ( for ECA?109 cells, 1. 11 vs. 1. 72, 1. 40 vs. 2. 14, 0. 42 vs. 0. 66, respectively;for TE?13 cells, 0.41 vs. 0.46, 0.43 vs. 0.65, 0.40 vs. 0.71, respectively ( all P<0.05) . The sensitivity enhancement ratio (SER) of ECA?109 and TE?13 cells were 1.35 and 1.43, respectively. Flow cytometry showed that the apoptosis rate of ECA?109 and TE?13 cells in the combined group were significantly higher than those of the radiation group [ for ECA?109 cells, ( 41. 31 ± 1. 52)% vs. ( 9. 54 ± 0.52)%;for TE?13 cells, (46.28±0.28)% vs. (11.32±0.31)%, both P<0.01]. Western blotting showed that the expression levels of EGFR and DNA?PKcs were not significantly different in all groups ( all P>0.05) . Compared with those of the control group, p?EGFR and p?DNA?PKcs of the radiation group were significantly higher in both cell lines ( P<0.05) , and theγH2AX levels in the radiation group and medicine group were significantly higher than that of the control group ( P<0.05) . Compared with those of the radiation group and medicine group, p?EGFR and p?DNA?PKcs protein expression in the combined group were decreased significantly (P<0.05), while γH2AX protein expression was significantly increased (P<0.05). Conclusions Nimotuzumab can enhance the radiosensitivity of esophageal cancer ECA?109 and TE?13 cells. The potential mechanism may be related to the inhibition of EGFR phosphorylation and down?regulation of DNA damage repair proteins. The radiosensitizing effect of nimotuzumab is greater on poorly differentiated esophageal cancer cells.