AIM: To explore the clinical application effect of visual training equipment combined with conventional corrective treatment on children with ametropic amblyopia(AMA).METHODS: Prospective randomized control study. A total of 188 children(376 eyes)with AMA treated in our hospital from June 2021 to December 2022 were selected, and they were divided into two groups using a random number table. The conventional correction group(94 cases, 188 eyes)received conventional correction treatment, while the visual training group(94 cases, 188 eyes)received visual training equipment combined with conventional correction treatment, both lasted for 12 mo. The best corrected visual acuity, diopter, eye accommodation function, adverse reactions, amblyopia recurrence rates, and clinical efficacy were compared between the two groups at 6 and 12 mo after treatment.RESULTS:The two groups each had 8 cases(16 eyes)detached, the rate of loss to follow-up was 8.5%, and 86 cases(172 eyes)were included in each group. There were statistically significant differences in the best corrected visual acuity, diopter, amplitude of accommodation, accommodation facility and accommodative lag between the two groups of children before and after treatment(all P<0.05). The total effective rate of the visual training group(98.8%)was higher than that of the conventional correction group(91.9%; P<0.05). There was no statistically significant difference in the total effective rate of clinical efficacy between the two groups in different age groups and different degrees of amblyopia(all P>0.05). There was no statistically significant difference in the incidence of redness and swelling between the two groups(P>0.05). The recurrence rate of amblyopia in the visual training group(1.2%)was lower than that in the conventional correction group(8.1%; P<0.05).CONCLUSION: The combination of visual training equipment and conventional correction therapy has a significant clinical effect on children with AMA, which can effectively correct visual acuity, adjusting diopter and improve eye accommodation function, and recurrence rate of amblyopia is low and safety is high.

ObjectiveTo investigate the effect and molecular mechanism of exosomes derived from bone marrow mesenchymal stem cells（BMSC-exos） modified with Bushen Yisui capsule（BSYS）-containing serum on promoting the differentiation and maturation of OLN-93 oligodendrocytes by regulating miR-15b/Wnt signaling pathway. MethodsOLN-93 cells were divided into 5 groups， including the normal（NC） group， BMSC-exos group， BSYS-BMSC-exos group， BSYS-BMSC+LV-miR-15b-5p inhibitor-exos group， and BSYS-BMSC+LV-miR-15b-5p NC-exos group. DiR staining was used to observe the uptake of Exos by OLN-93 cells. The effective dosage of BSYS-BMSC-exos on OLN-93 cells was assessed by cell proliferation and activity assay（CCK-8）. Stable BMSCs lentiviral transfection strains were established to inhibit miR-15b-5p expression in both BMSCs and their exos， and transfection efficiency was verified by real-time fluorescent quantitative polymerase chain reaction（Real-time PCR） detection of miR-15b-5p. The expressions of 2′，3′-cyclic nucleotide 3′-phosphodiesterase（CNPase） and myelin proteolipid protein（PLP） in OLN-93 cells were detected by immunocytochemistry（ICC） and Western blot. The mRNA expressions of miR-15b-5p and Wnt3a in OLN-93 cells were detected by Real-time PCR， and the protein expression of Wnt3a was measured by Western blot. The expression levels of key molecules in the Wnt/β-catenin signaling pathway of OLN-93 cells， including glycogen synthase kinase（GSK）-3β， β-catenin， and T-cell specific transcription factor 4/transcription factor 7-like 2（TCF4/TCF7L2）， were measured by Real-time PCR and Western blot. ResultsDiR-labeled Exos were efficiently taken up by OLN-93 cells. The CCK-8 assay results indicated that 20 mg·L^-1 of BSYS-BMSC-exos exhibited the most significant effect in enhancing OLN-93 cell viability（P<0.01） and this dosage was selected for subsequent experiments. Following lentiviral transfection of BMSCs， Real-time PCR results revealed that miR-15b-5p was significantly suppressed in BMSCs（P<0.01）， and miR-15b-5p was also notably inhibited in BSYS-BMSC-exos（P<0.01）. ICC analysis further revealed an increase in the number of differentiated， mature CNPase and PLP-positive cells following BSYS-BMSC-exos treatment（P<0.01）. Western blot results demonstrated that the protein expression of CNPase and PLP was significantly enhanced with BSYS-BMSC-exos treatment（P<0.01）. Additionally， BSYS-BMSC-exos also increased the expression levels of miR-15b-5p and p-β-catenin proteins in OLN-93 cells， while decreased the mRNA and protein expressions of Wnt3a， as well as the mRNA expressions of β-catenin and TCF4/TCF7L2， and the protein expression level of p-GSK-3β（Ser9） was significantly reduced（P<0.05， P<0.01）. After the transfection of miR-15b-5p inhibitor into BSYS-BMSC-exos， the above effects were significantly diminished（P<0.05， P<0.01）. ConclusionBSYS-BMSC-exos facilitate the differentiation and maturation of OLN-93 cells， and its mechanism is related to the upregulation of miR-15b-5p in OLN-93 cells， which inhibits the expression of Wnt3a and thereby suppresses the Wnt signaling pathway.

The transition of cancer cells from epithelial state to mesenchymal state awarded hepatocellular carcinoma (HCC) stem cell properties and induced tumorigenicity, drug resistance, and high recurrence rate. Reversing the mesenchymal state to epithelial state by inducing mesenchymal-epithelial remodeling could inhibit the progression of HCC. Using high-throughput screening, chrysin was selected from natural products to reverse epithelial-mesenchymal transition (EMT) by selectively increasing CDH1 expression. The target identification suggested chrysin exerted its anti-HCC effect through covalently and specifically binding threonine 205 (Thr205) of alpha-enolase (ENO1). For the first time, we revealed that ENO1 bound β-catenin mRNA, and recruited YTHDF2 to identify the m6A modified β-catenin in the 3'-UTR region to degrade β-catenin mRNA. Eventually, the CDH1 gene expression was improved through the regulation of β-catenin mRNA. ENO1/β-catenin mRNA interaction might be a promising target for cellular plasticity reprogramming. Moreover, chrysin could mediate mesenchymal‒epithelial remodeling through increasing degradation of β-catenin mRNA by promoting the binding of ENO1 and β-catenin mRNA. To the best of our knowledge, chrysin is the first reported small molecule inducing β-catenin mRNA degradation through binding to ENO1. The water-soluble derivative of chrysin may be a natural product-derived lead compound for circumventing metastasis, recurrence, and drug resistance of HCC by mediating mesenchymal‒epithelial remodeling.

Traditional Chinese medicine (TCM) is an ancient medical system distinctive and effective in treating cancer, depression, coronavirus disease 2019 (COVID-19), and other diseases. However, the relatively abstract diagnostic methods of TCM lack objective measurement, and the complex mechanisms of action are difficult to comprehend, which hinders the application and internationalization of TCM. Recently, while breakthroughs have been made in utilizing methods such as network pharmacology and virtual screening for TCM research, the rise of machine learning (ML) has significantly enhanced their integration with TCM. This article introduces representative methodological cases in quality control, mechanism research, diagnosis, and treatment processes of TCM, revealing the potential applications of ML technology in TCM. Furthermore, the challenges faced by ML in TCM applications are summarized, and future directions are discussed.

Porcine epidemic diarrhea virus (PEDV) is an intestinal coronavirus that can cause porcine epidemic diarrhea, leading to diarrhea, vomiting, weight loss, and even death in piglets. Due to the diversity of PEDV strains, traditional vaccines are difficult to sustainably and effectively prevent and control PEDV. This article reviews the strategies and mechanisms of active substances in regulating intracellular signaling pathways, viral proteins, and microbial metabolites to enhance the host immune function against PEDV. It emphasizes the prevention of PEDV resistance and the potential harm of PEDV breaking through interspecies barriers to the human society, aiming to provide reliable theoretical support for the development of new antiviral drugs or vaccines.
Porcine epidemic diarrhea virus/immunology* ; Animals ; Swine ; Swine Diseases/prevention & control* ; Antiviral Agents/pharmacology* ; Coronavirus Infections/virology* ; Viral Vaccines/immunology* ; Humans ; Signal Transduction

Objective:To investigate the clinical efficacy of different surgical approaches for repairing eyelid coloboma.Methods:Patients with the divided nevus of eyelid treated at Hypospadias Plastic Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, from January 2005 to January 2022 were included. The surgeries were categorized into 4 types. (1) Direct excision and suture. (2) Local skin grafts: covering the defect with split- or full-thickness skin grafts according to the size of the defect on the upper and lower eyelids. (3) Combined skin grafts with orbicularis oculi myocutaneous flap: grafting skin flaps for defects on the upper eyelid and near the lower eyelid, and temporal area skin flaps based on the same side orbicularis oculi muscle pedicle for lower eyelid defects. (4) Temporal area expanded flap based on the orbicularis oculi muscle combined with skin grafts: the surgery was divided into two stages, the first stage involves the placement of an expander in the temporal area of the affected side, and the second stage involves the removal of the expander, excision of eyelid lesion tissue and formation of an island-shaped skin flap with the orbital part of the orbicularis oculi muscle pedicle as the pedicle, which was rotated 180° to cover the lower eyelid defect. Defects near the upper and lower eyelid margins were still covered with skin grafts. Follow-up was conducted through outpatient visits, telephone calls, and WeChat messaging to assess facial appearance postoperatively. SPSS 22.0 statistical software was used for analysis, and the measurement data were expressed as Mean±SD, the count data were expressed as percentage, and the comparison of preoperative and postoperative was calculated by aesthetic and functional status of facial soft-tissue deformities (A&F scores) within the group was performed by paired t-test, the difference was considered statistically significant at P<0.05. Results:A total of 34 patients were included; average age was 17.7±15.3 years. The size of the lesions varied from the smallest nevus measuring 0.3 cm × 0.2 cm and the largest measuring 14.0 cm × 14.0 cm. Direct excision with suturing was performed in 6 cases, among which 2 cases received simultaneous double eyelid surgery, the postoperative A&F score (4.54 ± 1.32) was higher than the preoperative (3.28 ± 0.98) score, the difference was not statistically significant ( P>0.05). The skin graft was performed in 10 cases, the postoperative A&F score (5.13 ± 1.59) was higher than the preoperative (2.25 ± 1.59), the difference was not statistically significant ( P>0.05). The skin graft combined with using orbicularis oculi muscle skin flap was performed in 7 cases, the postoperative A&F score of our patients (5.54 ±1.46) was significantly higher than the preoperative (2.18 ±1.61 ), the difference was statistically significant ( P<0.05). The skin graft combined with the expanded temporal area flap based on the orbicularis oculi muscle was performed in 11 cases, the A&F score of our patients after repair (4.95 ±0.60) was improved compared with the preoperative (2.18±1.48) score, and the difference was not statistically significant ( P>0.05). Twenty-eight patients were followed up for 8 to 81 months while 6 cases lost due to change of contact information, among whom three developed secondary ectropion of the lower eyelid, three experienced recurrence of nevi in the incision and grafting area, two exhibited significant pigmentation in the grafting area, and one developed secondary ptosis of the upper eyelid, while the rest of the patients were satisfied with the repair results. Conclusion:Surgical excision is the only effective treatment for eyelid cleft nevi with different clinical presentations. Covering defects on the upper and lower eyelids with different tissue transplantation method can disperse the entire nevus, resulting in more stable long-term repair effects and a more natural appearance. The combination of an orbicularis muscle flap and a free skin graft provides more stable result in the plastic surgery of medium to large eyelid split nevi. Free skin grafting of the upper and lower lids near the margins, and flap grafting of the lower lids are more consistent with the physiological state of the upper and lower lids, and the combination of different tissue grafting method can make the appearance of the face more reasonable.

Objective:To investigate the clinical efficacy of different surgical approaches for repairing eyelid coloboma.Methods:Patients with the divided nevus of eyelid treated at Hypospadias Plastic Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, from January 2005 to January 2022 were included. The surgeries were categorized into 4 types. (1) Direct excision and suture. (2) Local skin grafts: covering the defect with split- or full-thickness skin grafts according to the size of the defect on the upper and lower eyelids. (3) Combined skin grafts with orbicularis oculi myocutaneous flap: grafting skin flaps for defects on the upper eyelid and near the lower eyelid, and temporal area skin flaps based on the same side orbicularis oculi muscle pedicle for lower eyelid defects. (4) Temporal area expanded flap based on the orbicularis oculi muscle combined with skin grafts: the surgery was divided into two stages, the first stage involves the placement of an expander in the temporal area of the affected side, and the second stage involves the removal of the expander, excision of eyelid lesion tissue and formation of an island-shaped skin flap with the orbital part of the orbicularis oculi muscle pedicle as the pedicle, which was rotated 180° to cover the lower eyelid defect. Defects near the upper and lower eyelid margins were still covered with skin grafts. Follow-up was conducted through outpatient visits, telephone calls, and WeChat messaging to assess facial appearance postoperatively. SPSS 22.0 statistical software was used for analysis, and the measurement data were expressed as Mean±SD, the count data were expressed as percentage, and the comparison of preoperative and postoperative was calculated by aesthetic and functional status of facial soft-tissue deformities (A&F scores) within the group was performed by paired t-test, the difference was considered statistically significant at P<0.05. Results:A total of 34 patients were included; average age was 17.7±15.3 years. The size of the lesions varied from the smallest nevus measuring 0.3 cm × 0.2 cm and the largest measuring 14.0 cm × 14.0 cm. Direct excision with suturing was performed in 6 cases, among which 2 cases received simultaneous double eyelid surgery, the postoperative A&F score (4.54 ± 1.32) was higher than the preoperative (3.28 ± 0.98) score, the difference was not statistically significant ( P>0.05). The skin graft was performed in 10 cases, the postoperative A&F score (5.13 ± 1.59) was higher than the preoperative (2.25 ± 1.59), the difference was not statistically significant ( P>0.05). The skin graft combined with using orbicularis oculi muscle skin flap was performed in 7 cases, the postoperative A&F score of our patients (5.54 ±1.46) was significantly higher than the preoperative (2.18 ±1.61 ), the difference was statistically significant ( P<0.05). The skin graft combined with the expanded temporal area flap based on the orbicularis oculi muscle was performed in 11 cases, the A&F score of our patients after repair (4.95 ±0.60) was improved compared with the preoperative (2.18±1.48) score, and the difference was not statistically significant ( P>0.05). Twenty-eight patients were followed up for 8 to 81 months while 6 cases lost due to change of contact information, among whom three developed secondary ectropion of the lower eyelid, three experienced recurrence of nevi in the incision and grafting area, two exhibited significant pigmentation in the grafting area, and one developed secondary ptosis of the upper eyelid, while the rest of the patients were satisfied with the repair results. Conclusion:Surgical excision is the only effective treatment for eyelid cleft nevi with different clinical presentations. Covering defects on the upper and lower eyelids with different tissue transplantation method can disperse the entire nevus, resulting in more stable long-term repair effects and a more natural appearance. The combination of an orbicularis muscle flap and a free skin graft provides more stable result in the plastic surgery of medium to large eyelid split nevi. Free skin grafting of the upper and lower lids near the margins, and flap grafting of the lower lids are more consistent with the physiological state of the upper and lower lids, and the combination of different tissue grafting method can make the appearance of the face more reasonable.

Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.

Developing and implementing biosafety standards for pathogenic microbiology laboratories is essential to achieving scientific, efficient, and standardized management and operation. This article analyzes the current standardization construction in biosafety in pathogenic microbiology laboratories domestically and internationally. It proposes a framework for the biosafety standard system of pathogenic microbiology laboratories, which mainly includes four parts: basic standards, management standards, technical standards, and industry applications. It provides a reference for the standardization work of pathogenic microbiology laboratories and helps to standardize the biosafety industry in China.

Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.