Objective To investigate the effect of E2 signaling in myofibers on muscular macrophage efferocytosis in mice with cardiotoxin-induced acute skeletal muscle injury.Methods Female wild-type C57BL/6 mice with and without ovariectomy and male C57BL/6 mice were given a CTX injection into the anterior tibial muscle to induce acute muscle injury,followed by intramuscular injection of β-estradiol(E2)or 4-hydroxytamoxifen(4-OHT).The changes in serum E2 of the mice were detected using ELISA,and the number,phenotypes,and efferocytosis of the macrophages in the inflammatory exudates and myofiber regeneration and repair were evaluated using immunofluorescence staining and flow cytometry.C2C12 cells were induced to differentiate into mature myotubes,which were treated with IFN-γ for 24 before treatment with β-Estradiol or 4-OHT.The treated myotubes were co-cultured with mouse peritoneal macrophages in a 1:2 ratio,followed by addition of PKH67-labeled apoptotic mouse mononuclear spleen cells induced by UV irradiation,and macrophage efferocytosis was observed using immunofluorescence staining and flow cytometry.Results Compared with the control mice,the female mice with ovariectomy showed significantly increased mononuclear macrophages in the inflammatory exudates,with increased M1 cell percentage,reduced M2 cell percentage and macrophage efferocytosis in the injured muscle,and obviously delayed myofiber regeneration and repair.In the cell co-culture systems,treatment of the myotubes with β-estradiol significantly increased the number and proportion of M2 macrophages and macrophage efferocytosis,while 4-OHT treatment resulted in the opposite changes.Conclusion In injured mouse skeletal muscles,myofiber E2 signaling promotes M1 to M2 transition to increase macrophage efferocytosis,thereby relieving inflammation and promoting muscle regeneration and repair.

Objective To investigate the effect of the glutathione peroxidase 4(GPx4)on mouse so-matic cell reprogramming.Methods To compare the expressions of GPx4 in OG2 mouse embryonic fibroblast(OG2-MEF)cells(MEFs group)and mouse embryonic stem cells(mESC,mESCs group),the expression lev-el of intracellular GPx4 was determined by transcriptome sequencing technique and Western blot.To verify the effect of GPx4 on the efficiency of the somatic cells reprogramming,the complete open reading frame se-quence of GPx4 gene and its selenocysteine insertion sequence(SECIS)were connected to the retroviral vector pMXs for constructing the overexpressed plasmid pMXs-GPx4.Gpx4-targeting short hairpin RNA(shRNA)was synthesized and connected to pSUPER vector,GPx4 shRNA1 and GPx4 shRNA2 were constructed to knockdown GPx4 expression.The above plasmids were co-transfected with pMXs-Sox2,pMXs-Klf4 and pMXs-Oct4 into MEF cells for reprogramming induction to obtain the pMXs no-load control group(pMXs NC),pMXs GPx4 group,pSUPER no-load control group(pSUPER NC),GPx4 shRNA1 group and GPx4 shRNA2 group.The expressions of GPx4 gene and multifunctional marker genes Rex1,Sox2,Dappa3,Sall4,Oct4 and Nanog were detected by real-time fluorescence quantitative PCR.The induced pluripotent stem cells(iPSC)were detected by immunofluorescence staining;the number of iPSC clones generation was detected by alkaline phosphatase staining of pluripotent stem cells;the GPx4 protein expression was detected by Western blot.Results The mRNA and protein expression of GPx4 in the mESCs group was higher than that in the MEFs group;compared with the pMXs NC group,the expression level of GPx4 mRNA in the pMXs GPx4 group was significantly increased;compared with the pSUPER NC group,the GPx4 mRNA and protein levels in the GPx4 shRNA1 group and GPx4 shRNA2 group were decreased(P＜0.05);the iPSC clone number in the pMXs GPx4 group was higher than that in the pMXs NC group,but the difference was not statistically significant(P＞0.05).The number of iPSC clones in the GPx4 shRNA1 group and GPx4 shRNA2 group was significantly lower than that in the pSUPER NC group,and the difference was statistically significant(P＜0.05).After completing the reprogramming,compared with the original MEF cells,the expression levels of various pluripotent marker genes Rex1,Sox2,Dappa3,Sall4,Oct4 and Nanog in the generated iPSC of each group were increased.Conclusion GPx4 knockdown could inhibit the efficiency of somatic cell reprogram-ming,its generated induced pluripotent stem cells have the normal pluripotent gene expression ability.

Objective To investigate the effect of E2 signaling in myofibers on muscular macrophage efferocytosis in mice with cardiotoxin-induced acute skeletal muscle injury.Methods Female wild-type C57BL/6 mice with and without ovariectomy and male C57BL/6 mice were given a CTX injection into the anterior tibial muscle to induce acute muscle injury,followed by intramuscular injection of β-estradiol(E2)or 4-hydroxytamoxifen(4-OHT).The changes in serum E2 of the mice were detected using ELISA,and the number,phenotypes,and efferocytosis of the macrophages in the inflammatory exudates and myofiber regeneration and repair were evaluated using immunofluorescence staining and flow cytometry.C2C12 cells were induced to differentiate into mature myotubes,which were treated with IFN-γ for 24 before treatment with β-Estradiol or 4-OHT.The treated myotubes were co-cultured with mouse peritoneal macrophages in a 1:2 ratio,followed by addition of PKH67-labeled apoptotic mouse mononuclear spleen cells induced by UV irradiation,and macrophage efferocytosis was observed using immunofluorescence staining and flow cytometry.Results Compared with the control mice,the female mice with ovariectomy showed significantly increased mononuclear macrophages in the inflammatory exudates,with increased M1 cell percentage,reduced M2 cell percentage and macrophage efferocytosis in the injured muscle,and obviously delayed myofiber regeneration and repair.In the cell co-culture systems,treatment of the myotubes with β-estradiol significantly increased the number and proportion of M2 macrophages and macrophage efferocytosis,while 4-OHT treatment resulted in the opposite changes.Conclusion In injured mouse skeletal muscles,myofiber E2 signaling promotes M1 to M2 transition to increase macrophage efferocytosis,thereby relieving inflammation and promoting muscle regeneration and repair.

Objective:To explore the use of near-infrared autofluorescence probe (NIRAF-P) and its application in identifying parathyroid glands during surgery.Methods:A total of 68 patients undergoing thyroid surgery at Peking Union Medical College Hospital and Beijing Longfu Hospital between Dec. 2023 and Jun. 2024 were selected. During the operation, the near-infrared parathyroid gland detector was used to identify the parathyroid gland tissue to be tested, and histopathological examination was performed. The positive predictive value and accuracy of the near-infrared parathyroid gland detector were analyzed.Results:A total of 111 parathyroid glands were identified in 68 patients, and the positive predictive value and accuracy of the NIRAF-P were 95.5% and 94.6%, respectively.Conclusions:The NIRAF-P has high accuracy in identifying parathyroid glands. The standardized application of the NIRAF-P can help improve the efficiency of identifying parathyroid glands during surgery.

Abstract Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.
Humans ; Clostridioides/metabolism* ; Clostridioides difficile/metabolism* ; Bacterial Proteins/metabolism* ; Virulence ; Anti-Bacterial Agents/metabolism*

Objective:To explore whether deep learning could apply to recognize the recurrent laryngeal nerve in the video of unilateral axillary approach endoscopic thyroidectomy.Methods:Videos of endoscopic thyroidectomy via unilateral axillary approach in Peking Union Medical College Hospital from Jul. 1st, 2020 to May. 1st, 2021 were collected. Videos containing the recurrent laryngeal nerve were selected, and the outline of recurrent laryngeal nerve were marked by two senior thyroid surgeons and staffs. Data were divided into training set and test set in a ratio of 5:1, and classified into high, medium and low recognition group according to difficulty of recognizing the outline of the nerve. The neuron network was based on PSPNet combined with Resnet50. All data were analyzed by R (ver. 4.0.2) .Results:A total of 38 videos including 35,501 frames of pictures were included in this study. 29, 704 frames of 32 videos were in our training set and 5797 frames of 6 videos were in the test set. When the intersection over union (IOU) threshold is 0.1, the sensitivity and precision is 100.0%/92.1%, 95.8%/80.2% and 81.0%/80.6% in high, medium and low recognition group respectively. When the IOU threshold is 0.5, the sensitivity and precision is 92.6%/85.3%, 71.7%/60.5% and 38.1%/37.9% in high, medium and low recognition group respectively, indicating that neuron network could located the outline of recurrent laryngeal nerve in high and medium recognition group. False negatives were often due to small targets and unclear boundaries.Conclusion:Recurrent laryngeal nerve recognition based on deep learning is feasible and has potential application value in endoscopic thyroidectomy, which may help surgeons reduce the risk of accidental injury of recurrent laryngeal nerve and improve the safety of thyroidectomy.

Objective:To explore whether deep learning could apply to recognize the recurrent laryngeal nerve (RLN) in videos of endoscopic thyroidectomy (ETE) via breast approach.Methods:Videos of ETE via breast approach in Peking Union Medical College Hospital from Feb. 2020 to Aug. 2021 were collected. Videos containing RLN were selected, and the outline of RLN was marked by two thyroid surgeons. Then data were divided into a training set and a test set in a ratio of 5:1 and classified into the high and low difficulty group according to a senior thyroid surgeon’s opinion. Those pictures were input to D-LinkNet model. Precision, sensitivity and mean dice index was calculated.Results:A total of 46 videos including 153, 520 frames of pictures were included in this study. 131,039 frames of 39 videos were in the training set and 22,481 frames of 7 videos were in the test set. When the intersection over union threshold was 0.1, the sensitivity and precision was 92.9%/72.8% and 47.6%/54.9% in high and low recognition group, respectively. When the intersection over union threshold was 0.5, the sensitivity and precision turned to 85.8%/67.2% and 37.6%/43.5% in high and low difficulty group, respectively. Mean Dice index was 0.781 and 0.663 in high and low difficulty group, respectively.Conclusions:RLN recognition based on deep learning is feasible and has potential application value in ETE, which may help surgeons reduce the risk of accidental injury of RLN and improve the safety of thyroidectomy.

Objective:To summarize the experience and the clinical data of patients with primary hyperparathyroidism undergoing endoscopic parathyroidectomy.Methods:A total of 24 patients who underwent endoscopic parathyroidectomy for primary hyperparathyroidism in Peking Union Medical College Hospital during Feb. 2021 to May. 2022 were concluded in this study (20 cases of parathyroidectomy via axillary approach and 4 cases of parathyroidectomy via thoracic and breast approach) . The operation time, postoperative drainage, length of stay, level of parathyroid hormone and serum calcium of those patients were collected. Postoperative complications and recurrence of hyperparathyroidism were also observed.Results:The postoperative levels of serum parathyroid hormone and serum calcium were significantly reduced (over 50%) compared with preoperative level ( P<0.05) . The average operation time was (96±22) min (64-157 min) . The mean postoperative drainage volume was (47±16) ml on day 1, (46±11) ml on day 2, and (30±9) ml on day 3, respectively. The average length of postoperative hospital stay was (2.8±1.1) days (2-6 days) . In one case of parathyroidectomy via axillary approach, the operation was converted to open surgery because of the low position of lesion. Other cases completed endoscopic surgery and obtained satisfactory cosmetic results. There were no postoperative complications such as bleeding, permanent hoarseness, coughing while drinking water, or surgical site infection. The mean follow-up time was (7.4±4.2) months (1-16 months) . There was no obvious discomfort and no recurrence during follow-up. Conclusion:Endoscopic parathyroidectomy is safe and effective in the treatment of primary hyperparathyroidism, which can be used as a surgical option for patients with cosmetic requirements.

Objective:To investigate the application value of machine learning algorithms for gauze detection in laparoscopic pancreatic surgery.Methods:The retrospective and descriptive study was conducted. The 80 intact laparoscopic pancreatic surgery videos from Peking Union Medical College Hospital of Chinese Academy of Medical Sciences with timing of July 2017 to July 2020 were collected. The training set was used to train the neural network, and the test set was used to test the ability of neural network for gauze detection under different difficulties. Under the supervision of two superior doctors, videos that containing gauze were selected and classified according to recognition difficulty into three difficulty level including easy, normal and hard difficulty, and further divided based on random number method into training set with 61 videos and test set with 19 videos in a ratio of 3:1 roughly. The minimum enclosing rectangle of the gauze were marked frame by frame. All images were input to the neural network model for training after normalization and preprocessing. For every image, the output of neural network is the predicted minimum enclosing rectangle of gauze. The intersection over union >0.5 was identified as positive result. Observation indicators: (1) video annotation and classification; (2) test outcomes of neural network for test set.Count data were represented as absolute numbers or percentages.Results:(1) Video annotation and classification: a total of 26 893 frames of images form 80 videos were annotated, with 61 videos including 22 564 frames of images as the training set and 19 videos including 4 329 frames of images as the test set. Of the training set, 19 videos including 5 791 frames of images were classifed as easy difficulty, 38 videos including 15 771 frames of images were classifed as normal difficulty, 4 videos including 1 002 frames of images were classifed as hard difficulty, respectively. Of the test set, 4 videos including 1 684 frames of images were classifed as easy difficulty, 6 videos including 1 016 frames of images were classifed as normal difficulty, 9 videos including 1 629 frames of images were classifed as hard difficulty, respectively. (2) Test outcomes of neural network for test set: the overall sensitivity and accuracy of gauze detection by neural network in the test set were 78.471%(3 397/4 329) and 69.811%(3 397/4 866), respectively. The sensitivity and accuracy of gauze detection by neural network were 94.478%(1 591/1 684) and 83.168%(1 591/1 913) in easy difficulty test set. The sensitivity and accuracy of gauze detection by neural network were 80.413%(817/1 016) and 70.859%(817/1 153) in normal difficulty test set, 60.712%(989/1 629) and 54.944%(989/1 800)in hard difficulty test set. The frame rate reached more than or equally to 15 fps. The overall false negative rate and false positive rate of gauze detection by neural network in the test set were 21.529%(932/4 329) and 30.189%(1 469/4 866), respectively. The false negative was mainly due to the existence of blurred images, too small gauze exposure or blood immersion of gauze. The false positive was caused by the reflection of connective tissue or body fluids.Conclusion:The machine learning algorithms for gauze detection in laparoscopic pancreatic surgery is feasible, which could help medical staff identify gauze.