Small interfering RNA (siRNA) has a promising future in the treatment of ocular diseases due to its high efficiency, specificity, and low toxicity in inhibiting the expression of target genes and proteins. However, due to the unique anatomical structure of the eye and various barriers, delivering nucleic acids to the retina remains a significant challenge. In this study, we rationally design PACD, an A-B-C type non-viral vector copolymer composed of a hydrophilic PEG block (A), a siRNA binding block (B) and a pH-responsive block (C). PACDs can self-assemble into nanosized polymeric micelles that compact siRNAs into polyplexes through simple mixing. By evaluating its pH-responsive activity, gene silencing efficiency in retinal cells, intraocular distribution, and anti-angiogenesis therapy in a mouse model of hypoxia-induced angiogenesis, we demonstrate the efficiency and safety of PACD in delivering siRNA in the retina. We are surprised to discover that, the PACD/siRNA polyplexes exhibit remarkable intracellular endosomal escape efficiency, excellent gene silencing, and inhibit retinal angiogenesis. Our study provides design guidance for developing efficient nonviral ocular nucleic acid delivery systems.

BACKGROUND:m6A modification has been confirmed to play an important role in the occurrence and development of osteonecrosis of the femoral head;however,the role of m6A modification patterns in steroid-induced osteonecrosis of the femoral head remains unknown. OBJECTIVE:Bioinformatics analysis was performed based on the Gene Expression Omnibus(GEO)database to analyze the differential expression of the m6A gene in steroid-induced osteonecrosis of the femoral head,predict the downstream targeted miRNAs,and investigate the potential pathogenesis. METHODS:Expressing profiles of mRNA data of steroid-induced osteonecrosis of the femoral head were downloaded from GEO database(GSE123568).Differentially expressed genes(DEGs),Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using the R software.After obtaining these differentially methylated m6A genes(m6A-DEGs),we analyzed GO function and KEGG pathway enrichment and compared the correlation among the m6A-DEGs typing according to gene expression.The protein-protein interaction network and core gene subnetwork of m6A-DEGs were constructed using Cytoscape software.The m6A-DEGs-associated potential miRNAs were predicted using the TargetScan,miRTarBase,and miRBD databases.Simultaneously,ChIPBase and hTFtarget databases were used to predict potential transcription factors of seven core genes,then m6A-miRNA and transcription factor-m6A regulatory networks were constructed separately.Finally,the expression levels of the seven core m6A-DEGs were verified by using the GSE74089 dataset. RESULTS AND CONCLUSION:(1)A total of 2 460 common DEGs were screened out from datasets,among which 1 455 genes were upregulated and 1 005 genes were downregulated.(2)A total of 14 m6A-DEGs were identified in the datasets.Among them,11 m6A-DEGs were up-regulated and 3 m6A-DEGs were down-regulated.Differential gene expression was considered significant for m6A-DEGs in steroid-induced osteonecrosis of the femoral head(P<0.05).Spearman correlation analysis showed a significant correlation between m6A-DEGs.(3)GO and KEGG enrichment analysis showed that m6A-DEGs were mainly enriched in myeloid cell differentiation and development,immune and cytokine receptor activity,osteoclast differentiation,AMPK signaling pathway and interleukin-17 signaling pathway.(4)The seven core genes of m6A-DEGs contained YTHDF3,YTHDF1,YTHDF2,ALKBH5,METTL3,HNRNPA2B1,and HNRNPC.A total of 44 miRNAs overlapping were detected in the miRTarBase,miRDB,and TargetScan databases.Totally 79 transcription factors overlapping were found in the ChIPBase and hTFtarget databases.(5)The expression levels of six core m6A-DEGs in the GSE74089 dataset were consistent with those in the GSE123568 dataset.(6)These findings confirm that the seven m6A-DEGs identified through bioinformatics techniques play a regulatory role in the expression of various miRNAs,transcription factors,AMPK,and interleukin-17 signaling pathways,and these genes have a significant impact on the differentiation and development of bone marrow cells as well as osteoclast differentiation in steroid-induced osteonecrosis of the femoral head.Consequently,these findings offer data support and establish a research direction for future investigations into the pathogenesis and targeted therapeutic strategies for steroid-induced osteonecrosis of the femoral head.

AIM To explore the mechanism of Yiqi Wenyang Huwei Decoction on airway inflammation improvement of rats with bronchial asthma based on IL-25/NF-κB signaling pathway.METHODS 60 rats were randomly divided into the control group,the model group,the dexamethasone group(0.2 mg/mL),the low-dose,medium-dose and high-dose Yiqi Wenyang Huwei Decoction groups(1,2,4 g/mL),with 10 rats in each group.Intraperitoneal injection of ovalbumin(OVA)and aluminum hydroxide suspension was applied to establish the rat asthma model,followed by 2-week corresponding dosing of the drugs.The rats of each group had their daily diet,mental status,hair growth and respiration observed;their differential count of inflammatory cells in bronchoalveolar lavage fluid(BALF)detected by automatic hematology analyzer;their pathological changes of lung tissue observed by HE staining;their pulmonary IL-25 protein expression detected by immunohistochemistry(IHC);their levels of IL-4,IL-5 and IL-13 in BALF measured by ELISA;their pulmonary expression of IL-25 and TRAF6 mRNA detected by RT-qPCR;and their pulmonary protein expressions of IL-25,TRAF6,IκBα,p-IκBα,NF-κB p65 and p-NF-κB p65 detected by Western blot.RESULTS Compared with the control group,the model group displayed severe damage of the lung tissue and infiltration of a large number of inflammatory cells;increased number of inflammatory cells and levels of IL-4,IL-5 and IL-13 in BALF(P<0.01);increased mRNA expressions of IL-25 and TRAF6,and pulmonary protein expressions of IL-25,TRAF6,p-IκBα/IκBα and p-NF-κB p65/NF-κB p65(P<0.01).Compared with the model group,all of the Yiqi Wenyang Huwei Decoction groups shared improved pulmonary infiltration of inflammatory cells;decreased number of inflammatory cells and levels of IL-4,IL-5 and IL-13 in BALF(P<0.05,P<0.01);and decreased mRNA expressions of IL-25 and TRAF6,and pulmonary protein expressions of IL-25,TRAF6,p-IκBα/IκBα and p-NF-κB p65/NF-κB p65(P<0.01).CONCLUSION Yiqi Wenyang Huwei Decoction can inhibit the airway inflammation in the rat model of bronchial asthma,which may be related to the inhibited activation of IL-25/NF-κB signaling pathway and the reduced expression of inflammatory factors.

Objective:To introduce the application of highly selective arterial indocyanine green angiography (hereinafter referred to as highly selective arterial angiography) in the design of anterolateral thigh free flap.Methods:This study was a retrospective observational study. From November 2023 to April 2024, 29 patients with wounds in extremities which were repaired by anterolateral thigh free flaps designed under the assistance of highly selective arterial angiography and met the inclusion criteria were admitted to the Department of Hand Surgery and Department of Wound Repair Surgery of Suzhou Ruihua Orthopedic Hospital, including 26 males and 3 females, aged 16 to 71 years. The wound area after debridement ranged from 8.0 cm×4.5 cm to 27.0 cm×16.0 cm. During the surgery, highly selective arterial angiography was used to assist in flap design. The fluorescence development range of the source arteries or perforators of flaps was observed. The blood supply range of the source arteries or perforators of flaps was determined based on the fluorescence development of the skin, and the excision position of the flap was adjusted. The flap incision area ranged from 9.0 cm×6.0 cm to 29.0 cm×16.0 cm. During the surgery, the number of highly selective arterial angiography, the type of source artery of perforators for puncture, and changes in the excision position of flaps were observed and recorded. After surgery, the blood supply and survival of flaps, the healing of wounds and the survival of skin grafts in the flap donor sites, and the angiography-related complications were observed.Results:All the 32 flaps of 29 patients were successfully excised. The highly selective arterial angiography was performed 37 times, including 13 cases of puncture of the oblique branch of the lateral circumflex femoral artery, 6 cases of puncture of the descending branch, 8 cases of double puncture of the oblique and descending branches, and 2 cases of puncture of arteries from other branches. During the surgery, the excision position of 28 flaps did not change, the excision position of 3 flaps moved towards proximal extremity of the thigh, and the excision position of 1 flap moved towards distal extremity of the thigh. All the flaps survived successfully after the surgery, and there was no partial necrosis of the flaps at the proximal or distal ends. The wounds in the flap donor sites healed, and all skin grafts survived. No angiography-related complications occurred.Conclusions:Highly selective arterial angiography can be used to determine the blood supply range of the source artery and perforators of the anterolateral thigh free flaps during the surgery. It can evaluate the blood supply of flaps more intuitively and objectively. Its application in assisting flap design can avoid partial flap necrosis caused by unreasonable preoperative design to a certain extent, and it is safe and reliable.

Objective:To compare the clinical efficacies of O-arm combined with CT three-dimensional navigation system assisted screw placement versus manual screw placement in treating lower cervical fracture and dislocation.Methods:A retrospective cohort study was used to analyze the clinical data of 41 patients with lower cervical fracture and dislocation, who were treated in Honghui Hospital, Xi′an Jiaotong University from May 2021 to February 2022. The patients included 26 males and 15 females, aged 31.5-48.6 years [(41.5±15.0)years]. The injured segments were C 3 in 3 patients, C 4 in 12, C 5 in 13, C 6 in 10 and C 7 in 3. Nineteen patients were treated with cervical pedicle screws by O-shaped arm combined with CT three-dimensional navigation system (navigation group, 76 screws) and 22 by bare hands (traditional group, 88 screws). The total operation time, effective operation time, single nail placement time, single screw correction times, screw distance from anterior cortex, intraoperative blood loss, intraoperative fluoroscopic radiation dose, incision length and length of hospital stay were compared between the two groups, and the height of intervertebral space, Cobb angle, interbody slip distance and American Spinal injury Association (ASIA) grade were compared before operation and at 3 days after operation. Visual analogue score (VAS), Japanese Orthopedic Association (JOA) score, and neck dysfunction index (NDI) were evaluated before operation, at 3 days, 3 months after operation and at the last follow-up. Accuracy of screw placement and incidence of complications (adjacent facet joint invasion, infection, screw loosening) were detected as well. Results:All the patients were followed up for 11.1-13.9 months [(12.5±1.4)months]. The total operation time, intraoperative blood loss, intraoperative fluoroscopic radiation dose and incision length in the navigation group were more or longer than those in the traditional group (all P<0.05). The effective operation time, single nail placement time, single nail correction times and screw distance from anterior cortex in the navigation group were markedly less or smaller than those in the traditional group (all P<0.05). There was no significant difference in the length of hospital stay between the two groups ( P>0.05). There were significant improvements in the height of intervertebral space, Cobb angle and interbody slip distance between the two groups at 3 days after operation (all P<0.05). There was no significant difference in the height of intervertebral space, Cobb angle, interbody slip distance or ASIA grade between the two groups before operation or at 3 days after operation (all P>0.05). Compared with pre-operation, the VAS, JOA score and NDI were significantly improved in both groups at 3 days, 3 months after operation and at the last follow-up (all P<0.05), with further improvement with time. There was no significant difference in VAS between the two groups before operation or at 3 months after operation (all P>0.05), but it was markedly lower in the navigation group compared with the traditional group at 3 days after operation and at the last follow-up (all P<0.05). There were no significant differences in JOA score or NDI between the two groups before operation or at 3 days and 3 months after operation (all P>0.05), but both were lower in the navigation group compared with the traditional group at the last follow-up (all P<0.05). The accuracies of placement of grade 0 and grade 0+1 screws were 92.0% (70/76) and 96.6% (73/76) in the navigation group, respectively, which were markedly higher than 88.7% (78/88) and 93.5% (82/88) in the traditional group (all P<0.05). The rates of adjacent facet joint invasion of A, B, and C degrees were 71.2% (54/76), 28.8% (22/76) and 0% (0/76) in the navigation group, respectively, while the invasion rates were 60.5% (53/88), 32.3% (28/88) and 7.3% (7/88) in the traditional group ( P<0.05). No screw loosening was noted in the navigation group, but the screw loosening rate was 9.1% (8/88) in the traditional group ( P<0.01). Conclusion:Compared with manual screw placement, O-arm combined with CT three-dimensional navigation system assisted screw placement for lower cervical fracture and dislocation has the advantages of shorter effective operation time, quicker screw placement, stronger screw holding force, better cervical stability, slighter postoperative pain, higher screw placement accuracy, and lower facet joint invasion and screw loosening rates.

Age-dependent loss of skeletal muscle mass and function is a feature of sarcopenia, and increases the risk of many aging-related metabolic diseases. Here, we report phenotypic and single-nucleus transcriptomic analyses of non-human primate skeletal muscle aging. A higher transcriptional fluctuation was observed in myonuclei relative to other interstitial cell types, indicating a higher susceptibility of skeletal muscle fiber to aging. We found a downregulation of FOXO3 in aged primate skeletal muscle, and identified FOXO3 as a hub transcription factor maintaining skeletal muscle homeostasis. Through the establishment of a complementary experimental pipeline based on a human pluripotent stem cell-derived myotube model, we revealed that silence of FOXO3 accelerates human myotube senescence, whereas genetic activation of endogenous FOXO3 alleviates human myotube aging. Altogether, based on a combination of monkey skeletal muscle and human myotube aging research models, we unraveled the pivotal role of the FOXO3 in safeguarding primate skeletal muscle from aging, providing a comprehensive resource for the development of clinical diagnosis and targeted therapeutic interventions against human skeletal muscle aging and the onset of sarcopenia along with aging-related disorders.
Animals ; Humans ; Sarcopenia/metabolism* ; Forkhead Box Protein O3/metabolism* ; Muscle, Skeletal/metabolism* ; Aging/metabolism* ; Primates/metabolism*

The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.

Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.
Adult ; Humans ; Adolescent ; Imatinib Mesylate/adverse effects* ; Incidence ; Antineoplastic Agents/adverse effects* ; Retrospective Studies ; Pyrimidines/adverse effects* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy* ; Treatment Outcome ; Benzamides/adverse effects* ; Leukemia, Myeloid, Chronic-Phase/drug therapy* ; Aminopyridines/therapeutic use* ; Protein Kinase Inhibitors/therapeutic use*

Humans ; Mpox (monkeypox) ; China

Objective:To compare the efficacy of cervical decompression performed at different times in the treatment of incomplete cervical spinal cord injury.Methods:A multicenter retrospective cohort study was conducted to analyze the clinical data of 96 patients with incomplete cervical spinal cord injury admitted to six hospitals including Honghui Hospital affiliated to Xi'an Jiaotong University, etc, from May 2018 to May 2021. There were 36 females and 60 males, aged 28-42 years [(35.2±6.7)years]. The injured segments were at C 3 in 7 patients, C 4 in 15, C 5 in 20, C 6 in 23 and C 7 in 31. According to the American Spinal Injury Association (ASIA) scale, there were 59 patients with grade B, 27 grade C, and 10 grade D. A total of 36 patients underwent cervical decompression within 24 hours after injury (early group), 33 patients within 24-72 hours after injury (late group), and 27 patients within 4-14 days after injury (delayed group). The operation time, intraoperative blood loss, postoperative drainage, length of hospital stay, Cobb angle, height of intervertebral space and space occupation of the spinal canal before surgery and at postoperative 3 days, and ASIA score, ASIA motor score, ASIA light tactile score, ASIA acupuncture sensation score, visual analog scale (VAS) score, Japanese Orthopedic Association (JOA) score, neck dysfunction index (NDI) before surgery and at postoperative 3 months, 1 year and at the last follow-up and incidence of complications were compared among the three groups. Results:All the patients were followed up for 12-21 months [(16.4±4.2)months]. There was no significant difference in the operation time among the three groups (all P>0.05). The intraoperative blood loss and postoperative drainage volume in the early group were (312.5±5.2)ml and (165.3±45.8)ml, which were higher than those in the late group [(253.5±40.0)ml, (120.4±60.6)ml] and the delayed group [(267.3±36.8)ml and (130.4±38.6)ml] (all P<0.01). There was no significant difference between the late group and the delayed group (all P>0.05). The length of hospital stay in the early group was (5.2±1.6)days, which was shorter than that in the late group [(7.6±2.3)days] and the delayed group [(8.0±1.3)days] (all P<0.05), but there was no significant difference between the late group and the delayed group ( P>0.05). There was no significant difference in the Cobb angle, height of intervertebral space and space occupation of the spinal canal among the three groups before and at postoperative 3 days (all P>0.05). There was no significant difference in the ASIA score, ASIA motor score, ASIA light tactile score, ASIA acupuncture sensation score, VAS score, JOA score and NDI among the three groups before surgery (all P>0.05). At postoperative 3 months, 1 year and at the last follow-up, the ASIA grading of the early group was better than that of the late group and the delayed group ( P<0.05 or 0.01), but there was no statistically significant difference between the late group and the delayed group (all P>0.05). The ASIA motor scores of the early group were (56.4±4.5)points, (76.3±3.6)points and (85.4±6.5)points at postoperative 3 months, postoperative 1 year and the last follow-up, respectively, which were higher than those in the late group [(52.3±2.4)points, (60.3±8.6)points and (72.3±2.4)points] and the delayed group [(51.9±2.3)points, (62.8±4.6)points and (71.9±1.3)points]; the ASIA light tactile scores of the early group were (70.2±2.9)points, (72.6±4.3)points and (78.3±2.3)points, which were higher than those in the late group [(66.2±3.7)points, (68.3±1.6)points and (73.3±1.6)points] and the delayed group [(65.2±2.1)points, (67.8±1.9)points and (72.3±2.5)points]; acupuncture sensation scores of the early group were (71.9±3.1)points, (80.1±3.8)points and (89.1±7.6)points, which were higher than those in the late group [(67.4±2.7)points, (72.6±3.7)points and (77.9±1.8)points] and the delayed group [(68.3±2.2)points, (72.6±3.1)points and (77.2±1.9)points] (all P<0.05). VAS scores of the early group at postoperative 3 months, 1 year and at the last follow-up were (4.3±0.6)points, (2.4±0.3)points and (1.6±0.2)points, which were lower than those in the late group [(5.1±1.3)points, (4.1±0.6)points and (3.0±0.6)points] and the delayed group [(5.0±1.7)points, (4.0±0.8)points and (3.1±0.2)points]; JOA scores of the early group were (12.8±1.6)points, (14.4±2.6)points and (17.9±3.3)points, which were higher than those in the late group [(11.9±1.9)points, (13.3±1.6)points and (8.9±1.3)points] and the delayed group [(11.6±1.8)points, (13.2±1.4)points and (9.3±2.1)points]; NDI scores of the early group were 12.1±3.3, 10.1±2.1 and 7.3±1.4, which were lower than those in the late group (14.4±3.1, 12.3±1.6 and 8.9±1.3) and the delayed group (14.1±2.3, 12.9±1.9 and 9.5±2.1) (all P<0.05). There was no significant difference in all the above-mentioned scores at postoperative 3 months, 1 year and at the last follow-up between the late group and the delayed group (all P>0.05). The incidence of complications was 25.0% (9/36) in the early group, 27.3% (9/33) in the late group and 37.0% (10/27) in the delayed group (all P>0.05). Conclusion:Compared with within 24-72 hours and 4-14 days after injury, cervical decompression performed within 24 hours after injury for patients with incomplete cervical spinal cord injury can shorten the length of hospital stay, improve the function of the spinal cord nerves and relieve pain, with no increase of the incidence of complications.