Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H₂O₂ and divided into four groups: control (NC), 50 μmol/L H₂O₂, 110 μmol/L H₂O₂ and 150 μmol/L H₂O₂. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H₂O₂, and siSIRT3+H₂O₂. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O₂•⁻) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H₂O₂ decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O₂•⁻ did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O₂•⁻ level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H₂O₂ group exhibited decreased cellular ATP levels, elevated mitochondrial O₂•⁻ levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H₂O₂ group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.

Achondroplasia (ACH) is a common inherited skeletal dysplasia (inherited dwarfism) that compromises quality of life across the lifespan. In 2021, vosoritide became the first approved precision therapy for ACH and is now available in more than 40 countries. Compared with prior symptomatic measures, vosoritide has demonstrated favorable efficacy and a reassuring safety profile. Nevertheless, existing international ACH guidelines largely emphasize complication management and symptomatic care, and there is no unified consensus on pharmacologic therapy. To address this gap, an international expert group developed the International Consensus Guidelines for the Implementation and Monitoring of Vosoritide Therapy in Patients with Achondroplasia providing systematic recommendations that span the continuum of care - from initial patient contact and pre-treatment assessment to medication counseling, injection training, and long-term outcome monitoring. These recommendations complement and refine current management and nursing protocols for individuals with ACH and offer practical guidance for clinicians across diverse regions. This article highlights key elements of the guideline to provide evidence-based support and clinical direction for healthcare professionals in China treating children with ACH using vosoritide.
Humans ; Achondroplasia/drug therapy* ; Consensus ; Practice Guidelines as Topic ; Child

Hepatolenticular degeneration （HLD） is an autosomal recessive genetic disorder characterized by copper metabolism impairment， and it is one of the few treatable neurogenetic diseases. A retrospective analysis was performed for the clinical data of four patients with HLD who attended Beijing Ditan Hospital， Capital Medical University， from November 2023 to October 2024， and the genetic testing results of three patients were analyzed， along with a literature review. Among the four patients， there was one female patient and three male patients. All patients had the initial symptoms of dysarthria and limb tremors， and physical examination showed positive K-F rings， normal liver enzyme levels， a reduction in serum ceruloplasmin， and an increase in 24-hour urinary copper excretion. Three patients underwent ATP7B genetic testing， among whom two had compound heterozygous mutations， and one had a heterozygous mutation. In addition， two patients underwent cranial magnetic resonance imaging， and the results showed symmetrical long T1 and long T2 signals. All four patients received copper-chelating therapy and achieved a good outcome. This article reports the diagnosis and treatment of four patients with HLD and identifies a case of HLD resulting from a rare heterozygous mutation site， thereby expanding the variation spectrum of the ATP7B gene.

Objective:To investigate the expression and clinical significance of long noncoding RNA (lncRNA) AP006284.1 in bladder cancer tissues, and to analyze the regulatory effect and downstream mechanism of AP006284.1 knockdown on the proliferation and migration of bladder cancer cells. Methods:The expression of AP006284.1 in bladder cancer tissues, and the relationship between AP006284.1 expression and tumor stage and disease-free survival of bladder cancer patients were analyzed in the gene expression omnibus (GEO) database. AP006284.1 gene expression in bladder cancer cell lines, including MGH-U3, T24, UMUC-3, J82 and 5637, and bladder epithelial immortalized SV-HUC-1 cell were detected by real-time reverse transcription-PCR (RT-qPCR) assay. J82 cells were divided into the control group and the transfection group, and transfected with control plasmid and AP006284.1 knockdown plasmid, respectively. The effect of AP006284.1 knockdown on the proliferation of J82 cells was detected by RT-qPCR and cell counting kit-8 (CCK-8) assay. The effect of AP006284.1 knockdown on the migration of J82 cells was determined by the scratch healing assay. The target gene of AP006284.1 was predicted by LncRNA2Target and LncRNome databases. The target fragment of wild-type AP006284.1 ( AP006284.1-Wt) or mutant AP006284.1 ( AP006284.1-Mut) was constructed into pGL3 plasmid by dual luciferase gene reporter assay. J82 cells were co-transfected with miR-205-3p or miR-negtive control (miR-NC) to validate the targeting relationship between AP006284.1 and miR-205-3p. The correlation between miR-205-3p and AP006284.1 expression in bladder cancer tissues was further analyzed by the GEO database. The effect of AP006284.1 knockdown on the expression of miR-205-3p gene in J82 cells was detected by RT-qPCR assay. The effect of AP006284.1 knockdown on the expression of phosphorylated Ras protein (p-Ras), phosphorylated Raf protein (p-Raf), phosphorylated mitogen-activated protein kinase kinase (p-MEK), and phosphorylated extracellular signal-regulated kinase (p-ERK) of the ERK pathway was detected by Western blotting in J82 cells. Results:GEO database analysis showed that the relative expression of AP006284.1 in bladder cancer tissues ( n=304) was significantly higher than that in normal tissues ( n=28, P<0.01). The relative expression of AP006284.1 was positively correlated with the tumor stage of the bladder cancer patients ( P<0.01). Compared with bladder cancer patients with low expression of AP006284.1, patients with high expression had a lower disease-free survival ( P<0.01). Compared with the SV-HUC-1 cell (1.02±0.34), the expression level of AP006284.1 gene was upregulated in MGH-U3 cell (5.77±0.37), T24 cell (3.02±0.40), UMUC-3 cell (3.62±0.59), J82 cell (7.19±0.24) and 5637 cell (5.59±0.30) (all P<0.01). The expression level of the AP006284.1 gene was the highest in J82 cells, therefore, the J82 cells were selected for the study. The expression level of AP006284.1 gene in the control group (7.20±0.26) was 6.92 times higher than that in the transfection group (1.04±0.28, t=16.16, P<0.01). Compared with the control group (0.74±0.11, 1.35±0.09, 1.63±0.14, 1.74±0.11), the absorbance ( A) values of J82 cells in the transfection group (0.49±0.06, 0.95±0.14, 1.09±0.08, 1.13±0.11) were reduced than those in the control group at the 24, 36, 48 and 60 h after AP006284.1 knockdown (all P<0.05). The migration distance of J82 cells in the control group was significantly longer than that in the transfection group. The migration rate of the control group [(65.03±6.20)%], which was 2.58 times higher than that of the transfection group [(25.22±3.45)%, t=5.61, P<0.01]. The target site of miR-205-3p containing AP006284.1 was predicted by LncRNA2Target and LncRNome databases. Compared with miR-NC group (1.00±0.11), the relative activity of dual luciferase of AP006284.1-Wt gene was significantly downregulated in the miR-205-3p group (0.31±0.07, t=5.47, P<0.01). Compared with the miR-NC group (0.97±0.14), the relative activity of dual luciferase of AP006284.1-Mut vector (0.98±0.07) was not significantly change ( t=0.09, P>0.05). The GEO database analysis showed that the expression of AP006284.1 in bladder cancer tissues was negatively correlated with the expression of miR-205-3p ( P<0.01). The expression level of miR-205-3p gene in the transfection group (5.42±0.24) was 5.21 times higher than that in the control group (1.04±0.40, t=9.40, P<0.01). Compared with the control group (3.22±0.17, 5.56±0.19, 4.38±0.17, 5.74±0.36), the expressions of p-Ras (2.33±0.12), p-Raf (1.61±0.20), p-MEK (1.57±0.25), and p-ERK (2.40±0.28) of the ERK pathway were decreased in the transfected J82 cells (all P<0.01). Conclusions:AP006284.1 is highly expressed in bladder cancer tissues. Knockdown of AP006284.1 can inhibit the proliferation and migration of bladder cancer cells by regulating the miR-205-3p and ERK pathway proteins.

Objective : To construct a lentiviral overexpression cell line of HACE1 and to investigate its effects on the proliferation and migration of glioma cells． Methods : Using the bacterial liquid containing the full-length cDNA sequence of human HACE1 as a template，the lentiviral expression plasmid pCDH-HACE1 was constructed，and the lentiviral cell line was screened．Western blot was employed to detect the expression of HACE1 in glioma cells． Meanwhile，the CCK-8 assay and the cell scratch assay were utilized to assess the impacts of HACE1 on the prolif- eration and migration of glioma cells． Results : The lentiviral cell line with HACE1 overexpression was successfully established．Western blot analysis demonstrated that HACE1 could be effectively expressed in eukaryotic cells．The results of the CCK-8 assay showed that，compared with the control group，the cell proliferation in the HACE1 over- expression group was significantly enhanced at 24 hours and 48 hours after cultivation (both P＜0. 01) ．The cell scratch assay revealed that，at 24 hours and 48 hours after cultivation，the Scratch areas in the HACE1 overexpres- sion group were higher than those in the control group (P＜0. 05，P＜0. 01) ． Conclusion The lentiviral cell line with HACE1 overexpression is successfully constructed．HACE1 can promote the proliferation and migration abili- ties of U251 glioma cells，providing a foundation for further research．

Objective : To construct a lentiviral overexpression cell line of HACE1 and to investigate its effects on the proliferation and migration of glioma cells． Methods : Using the bacterial liquid containing the full-length cDNA sequence of human HACE1 as a template，the lentiviral expression plasmid pCDH-HACE1 was constructed，and the lentiviral cell line was screened．Western blot was employed to detect the expression of HACE1 in glioma cells． Meanwhile，the CCK-8 assay and the cell scratch assay were utilized to assess the impacts of HACE1 on the prolif- eration and migration of glioma cells． Results : The lentiviral cell line with HACE1 overexpression was successfully established．Western blot analysis demonstrated that HACE1 could be effectively expressed in eukaryotic cells．The results of the CCK-8 assay showed that，compared with the control group，the cell proliferation in the HACE1 over- expression group was significantly enhanced at 24 hours and 48 hours after cultivation (both P＜0. 01) ．The cell scratch assay revealed that，at 24 hours and 48 hours after cultivation，the Scratch areas in the HACE1 overexpres- sion group were higher than those in the control group (P＜0. 05，P＜0. 01) ． Conclusion The lentiviral cell line with HACE1 overexpression is successfully constructed．HACE1 can promote the proliferation and migration abili- ties of U251 glioma cells，providing a foundation for further research．

Objective To explore enteral nutrition support and analyze its influencing factors in elderly patients with ischemic stroke. Methods A total of 328 patients with ischemic stroke in General Hospital of Western Theater Command were enrolled for nutritional screening between July 2020 and February 2024. Corresponding nutritional support plans were selected to investigate the compliance of patients with enteral nutrition support. Patients were divided into a standard group (n=140) and a non-standard group (n=97) based on whether their calorie intake met the standard. The effects of different clinical characteristics on enteral nutrition support were explored, and logistic analysis was used to analyze the influencing factors of non-standard enteral nutrition support. Results In the 328 patients with ischemic stroke, proportions of total parenteral nutrition support, total enteral nutrition support, and parenteral/enteral nutrition support were 25.30%, 27.74% and 46.95%, respectively. The proportions of vomiting or regurgitation, gastric residual volume >100 mL, mechanical ventilation and use of antibiotics >2 in the non-standard group were higher than those in the standard group (P<0.05). Logistic analysis showed that the above clinical characteristics were risk factors influencing patients with enteral nutrition support and parenteral/enteral nutrition support. Conclusion Vomiting or regurgitation , gastric residual volume, mechanical ventilation, and amount of antibiotics used are important influencing factors of enteral nutrition support in patients. Clinicians should pay attention to the above clinical characteristics.

ObjectiveTo investigate the efficacy of Xingpi capsules （XPC） in treating diarrhea-predominant irritable bowel syndrome （IBS-D） with spleen deficiency and elucidate its potential molecular mechanisms. MethodsA rat model of IBS-D with spleen deficiency was established by administering senna leaf in combination with restrained stress and swimming fatigue for 14 d. Ten specific pathogen free （SPF）-grade healthy rats were used as the normal control group. After successful modeling， SPF-grade rats were randomly divided into a model group， a pinaverium bromide group （1.5 mg·kg^-1）， and low- and high-dose XPC groups （0.135 and 0.54 g·kg^-1）， with 10 rats in each group. Rats in the normal control group and the model group were given distilled water by gavage， while the remaining groups were administered corresponding drug solutions by gavage once a day for 14 consecutive days. The rat body weights and fecal condition were observed every day， and the Bristol score was recorded. Enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of 5-hydroxytryptamine （5-HT） in serum and colon tissue. Transmission electron microscopy was used to observe the microvilli and tight junctions in the colon. The integrity of the colonic barrier， intestinal motility， and expression of related pathway proteins were evaluated by hematoxylin-eosin （HE） staining， immunohistochemistry， and Western blot. ResultsCompared with those in the normal control group， rats in the model group showed a significantly decreased body weight and increased diarrhea rate， diarrhea grade， and Bristol score （P<0.01）. HE staining revealed incomplete colonic mucosa in the model group， with evident congestion and edema observed. Electron microscopy results indicated decreased density and integrity of the colonic barrier， shedding and disappearance of microvilli， and significant widening of tight junctions. The expression levels of colonic tight junction proteins Occludin and Claudin-5 were downregulated （P<0.01）， and the levels of 5-HT in serum and colon tissue were elevated （P<0.01）. The small intestine propulsion rate significantly increased （P<0.01）， and the expression of contractile proteins Ras homolog family member A （RhoA） and Rho-associated coiled-coil containing protein kinase 2 （ROCK2） in colon and phosphorylation of myosin light chain （MLC₂₀） were upregulated （P<0.01）. Compared with the model group， the treatment groups showed alleviated diarrhea， diarrhea-associated symptoms， and pathological manifestations of colon tissue to varying degrees. Specifically， high-dose XPC exhibited effectively relieved diarrhea， promoted recovery of colonic mucosal structure， significantly reduced congestion and edema， upregulated expression of Occludin and Claudin-5 （P<0.01）， decreased levels of 5-HT in serum and colon tissue （P<0.05，P<0.01）， significantly slowed small intestine propulsion rate （P<0.01）， and significantly downregulated expression of contractile proteins RhoA and ROCK2 in colon and phosphorylation of MLC₂₀ （P<0.05，P<0.01）. ConclusionXPC effectively alleviates symptoms of spleen deficiency and diarrhea and regulates the secretion of brain-gut peptide. The characteristics of XPC are mainly manifested in alleviating IBS-D with spleen deficiency from the aspects of protecting intestinal mucosa and inhibiting smooth muscle contraction， and the mechanism is closely related to the regulation of the 5-HT-RhoA/ROCK2 pathway expression.

ObjectiveTo investigate the regulatory effect of icariin （ICA） on transforming growth factor-β₁ （TGF-β₁）/Smad pathway in bone microvascular endothelial cells （BMECs） and the effect on autophagy in BMECs. MethodsBMECs were isolated and cultured， and the cell types were identified by immunofluorescence. Cells were divided into the control group， model group （0.1 g·L^-1 methyl prednisolone）， ICA group （0.1 g·L^-1 methyl prednisolone +1×10^-5 mol·L^-1 ICA）， and TGF-β inhibitor group （0.1 g·L^-1 methyl prednisolone +1×10^-5 mol·L^-1 ICA +1×10^-5 mol·L^-1 LY2157299）. Transmission electron microscopy was used to observe the ultrastructure and autophagosome number of BMECs. Autophagy double-standard adenovirus was used to monitor the confocal autophagy flow generation of each cell. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the gene and protein expression of autophagy in the TGF-β₁/ Smad pathway. ResultsAfter cell separation culture， platelet endothelial cell adhesion molecule （CD31） and von willebrand factor （vWF） immunofluorescence identified BMECs. Transmission electron microscopy showed that the cell membrane was damaged， and the nucleus was pyknotic and broken in the model group. Compared with the model group， the ICA group had complete cell membranes， clear structures， with autophagy-lysosome sparsely distributed. The confocal photo showed that BMECs had autophagosomes and autophagy-lysosomes， and the autophagy expression of the ICA group was similar to that of the blank group. Compared with the blank group， in the model group and the LY2157299 group， autophagosomes and autophagy-lysosomes were barely seen in the autophagy flow. Compared with the blank group， the mRNA and protein expressions of autophagy effector protein 1 （Beclin1） and microtubule-associated protein 1 light chain 3B （LC3B） in the model group were significantly decreased （P<0.01）， and those of ubiquitin-binding protein （p62） were significantly increased （P<0.01）. The mRNA expression of TGF-β₁， Smad homolog 2 （Smad2）， and Smad homolog 3 （Smad3） decreased （P<0.05， P<0.01）. The protein expressions of TGF-β₁， p-Smad2， and p-Smad3 were significantly decreased （P<0.01）. Compared with those of the model group， the mRNA and protein expression of Beclin1 and LC3B in BMECs of the ICA group increased （P<0.01）， and those of p62 significantly reduced （P<0.01）. The mRNA expression of TGF-β₁， Smad2， and Smad3 increased significantly （P<0.01）. The protein expression of TGF-β₁， p-Smad2， and p-Smad3 increased significantly （P<0.01）. Compared with those in the model group， the mRNA and protein expressions of Beclin1， LC3B， and p62 in the inhibitor group were not statistically significant. The expression of key genes and proteins of the TGF-β₁ pathway in the inhibitor group was not statistically significant. ConclusionICA can promote glucocorticoid-induced autophagy expression of BMECs， and its mechanism may be related to activating the TGF-β₁/Smad signaling pathway.

Steroid-induced osteonecrosis of the femoral head （SONFH） is a severe musculoskeletal disorder often induced by the prolonged or excessive use of glucocorticoids. Characterized by ischemia of bone cells， necrosis， and trabecular fractures， SONFH is accompanied by pain， femoral head collapse， and joint dysfunction， which can lead to disability in severe cases. The pathogenesis of SONFH involves hormone-induced osteoblast apoptosis， bone microvascular endothelial cell （BMEC） apoptosis， oxidative stress， and inflammatory responses. The phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway plays a pivotal role in the development of the disease. Modulating the PI3K/Akt signaling pathway can promote Akt phosphorylation， thereby stimulating the osteogenic differentiation of bone marrow mesenchymal stem cells and osteoblasts， promoting angiogenesis in BMECs， and inhibiting osteoclastogenesis. The research on the treatment of SONFH with traditional Chinese medicine （TCM） has gained increasing attention. Recent studies have shown that TCM monomers and compounds have potential therapeutic effect on SONFH by intervening in the PI3K/Akt signaling pathway. These studies not only provide a scientific basis for the application of TCM in the treatment of SONFH but also offer new ideas for the development of new therapeutic strategies. This review summarized the progress in Chinese and international research on the PI3K/Akt signaling pathway in SONFH over the past five years. It involved the composition and transmission mechanisms of the signaling pathway， as well as its regulatory effects on osteoblasts， mesenchymal stem cells， osteoclasts， BMECs， and other cells. Additionally， the review explored the TCM understanding of SONFH and the application of TCM monomers and compounds in the intervention of the PI3K/Akt pathway. By systematically analyzing and organizing these research findings， this article aimed to provide references and point out directions for the clinical prevention and treatment of SONFH and promote further development of TCM in this field. With in-depth research on the PI3K/Akt pathway and the modern application of TCM， it is expected to bring safer and more effective treatment options for patients with SONFH.