Investigating the molecular mechanism of Mori Cortex against osteosarcoma by bioinformatics analysis and in vitro experimental.

OBJECTIVE: To explore the therapeutic mechanism of Mori Cortex against osteosarcoma (OS), we conducted bioinformatics prediction followed by in vitro experimental validation. METHODS: Gene expression data from normal and OS tissues were obtained from the GEO database and underwent differential analysis. Active Mori Cortex components and target genes were extracted from the Traditional Chinese Medicine System Pharmacology database. By intersecting these targets with differentially expressed genes in OS, we identified potential drug action targets. Using the STRING database, a protein-protein interaction network was constructed. Subsequent analyses of these intersected genes, including Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment, were performed using R software to elucidate biological processes, molecular functions, and cellular components, resulting in the simulation of signaling pathways. Molecular docking assessed the binding capacity of small molecules to signaling pathway targets. In vitro validations were conducted on U-2 OS cells. The CCK8 assay was used to determine drug-induced cytotoxicity in OS cells, and Western Blotting was employed to validate the expression of AKT, extracellular signal-regulated kinases (ERK), Survivin, and Cyclin D1 proteins. RESULTS: Through differential gene expression analysis between normal and OS tissues, we identified 12,364 differentially expressed genes. From the TCSMP database, 39 active components and 185 therapeutic targets related to OS were derived. The protein-protein interaction network indicated that AKT1, IL-6, JUN, VEGFA, and CASP3 might be central targets of Mori Cortex for OS. Molecular docking revealed that the active compound Morusin in Mori Cortex exhibits strong binding affinity to AKT and ERK. The CCK8 assay showed that Morusin significantly inhibits the viability of U-2 OS cells. Western Blot demonstrated a reduction in the p-AKT/AKT ratio, the p-ERK/ERK ratio, Survivin, and Cyclin D1. CONCLUSION: Mori Cortex may exert its therapeutic effects on OS through multiple cellular signaling pathways. Morusin, the active component of Mori Cortex, can inhibit cell cycle regulation and promote cell death in OS cells by targeting AKT/ERK pathway.

Bioinformatics-based discovery of biomarkers and immunoinflammatory targets in children with cerebral palsy: An observational study.

Cerebral palsy (CP) is the most common disabling disease in children, and motor dysfunction is the core symptom of CP. Although relevant risk factors have been found to be closely associated with CP: congenital malformations, multiple gestation, prematurity, intrauterine inflammation and infection, birth asphyxia, thrombophilia, and perinatal stroke. Its important pathophysiological mechanism is amniotic fluid infection and intraamniotic inflammation leading to fetal developing brain damage, which may last for many years. However, the molecular mechanism of CP is still not well explained. This study aimed to use bioinformatics to identify key biomarker-related signaling pathways in CP. The expression profile of children with CP was selected from the Gene Expression Comprehensive Database, and the CP disease gene data set was obtained from GeneCards. A protein-protein interaction network was established and functional enrichment analysis was performed using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases. A total of 144 differential key intersection genes and 10 hub genes were identified through molecular biology. Gene Ontology functional enrichment analysis results show that differentially expressed genes are mainly concentrated in biological processes, such as immune response and neurogenesis. The cellular components involved mainly include axons, postsynaptic membranes, etc, and their molecular functions mainly involve proteoglycan binding, collagen binding, etc. Kyoto Encyclopedia of Genes and Genomes analysis shows that the intersection genes are mainly in signaling pathways related to the immune system, inflammatory response, and nervous system, such as Th17 cell differentiation, Toll-like receptor signaling pathway, tumor necrosis factor signaling pathway, NF-κB signaling pathway, axon guidance, PI3K-Akt signaling pathway, HIF-1 signaling pathway, gap junction, etc. Jak-STAT signaling pathway, mTOR signaling pathway, and related hub genes regulate immune cells and inflammatory factors and play an important role in the development and progression of CP.

Mefenamic acid exhibits antitumor activity against osteosarcoma by impeding cell growth and prompting apoptosis in human osteosarcoma cells and xenograft mice model.

The study investigates the anticancer activity of mefenamic acid against osteosarcoma, shedding light on its underlying mechanisms and therapeutic potential. Mefenamic acid exhibited robust inhibitory effects on the proliferation of MG-63, HOS, and H2OS osteosarcoma cells in a dose-dependent manner. Moreover, mefenamic acid induced cellular toxicity in MG63 cells, as evidenced by LDH leakage, reflecting its cytotoxic impact. Furthermore, mefenamic acid effectively suppressed the migration and invasion of MG-63 cells. Mechanistically, mefenamic acid induced apoptosis in MG-63 cells through mitochondrial depolarization, activation of caspase-dependent pathways, and modulation of the Bcl-2/Bax axis. Additionally, mefenamic acid promoted autophagy and inhibited the PI3K/Akt/mTOR pathway, further contributing to its antitumor effects. The molecular docking studies provide compelling evidence that mefenamic acid interacts specifically and strongly with key proteins in the PI3K/AKT/mTOR pathway, suggesting a novel mechanism by which mefenamic acid could exert anti-osteosarcoma effects. In vivo studies using a xenograft mouse model demonstrated significant inhibition of MG-63 tumor growth without adverse effects, supporting the translational potential of mefenamic acid as a safe and effective therapeutic agent against osteosarcoma. Immunohistochemistry staining corroborated the in vivo findings, highlighting mefenamic acid's ability to suppress tumor proliferation and inhibit the PI3K/AKT/mTOR pathway within the tumor microenvironment. Collectively, these results underscore the promising therapeutic implications of mefenamic acid in combating osteosarcoma, warranting further investigation for clinical translation and development.

Analysis of the relationship between color and natural pigments of tobacco leaves during curing.

Color is one of the most important indicators for the flue-cured tobacco quality. The color change of tobacco has a great relationship with the natural pigments in the tobacco. The relationship between color characteristics and the content of natural pigments in tobacco leaves during curing was investigated. The middle part of variety K326 tobacco was taken at each key time point during the curing process to determine the changes of color characteristics, moisture, pigment and polyphenol content. The results showed that moisture content of wet basis of tobacco gradually decreased from 72 to 18% during the curing process, the b* value increased and then decreased, and the a* value increased significantly. The lutein and ß-carotene content decreased to 63.83 µg/g and 28.3 µg/g, respectively. The total polyphenols content increased to 50.19 mg/g. Meanwhile, the a* value was significantly and positively correlated with polyphenols content and negatively correlated with pigments content. Cluster analysis showed that the samples were divided into three categories: samples with the curing time of 0 h, 24-72 h, and 84-132 h. These results demonstrated that the color change of tobacco during curing process can be divided into three stages from the perspective of chemical composition, which are strongly related to the degradation of pigments and the transformation of polyphenols.

Research progress of small-molecule drugs in targeting telomerase in human cancer and aging.

Telomeres are unique structures located at the ends of linear chromosomes, responsible for stabilizing chromosomal structures. They are synthesized by telomerase, a reverse transcriptase ribonucleoprotein complex. Telomerase activity is generally absent in human somatic cells, except in stem cells and germ cells. Every time a cell divides, the telomere sequence is shortened, eventually leading to replicative senescence and cell apoptosis when the telomeres reach a critical limit. However, most human cancer cells exhibit increased telomerase activity, allowing them to divide continuously. The importance of telomerase in cancer and aging has made developing drugs targeting telomerase a focus of research. Such drugs can inhibit cancer cell growth and delay aging by enhancing telomerase activity in telomere-related syndromes or diseases. This review provides an overview of telomeres, telomerase, and their regulation in cancer and aging, and highlights small-molecule drugs targeting telomerase in these fields.

Corylin inhibits the progression of Non-small cell lung cancer cells by regulating NF-κB signaling pathway via targeting p65.

BACKGROUND: Lung cancer is characterized by high-risk and high mortality, among which non-small cell lung cancer (NSCLC) conquers a dominant position. Previous studies have reported that corylin has anti-inflammatory, anti-oxidant, and anti-tumor effects; however, its role in NSCLC cells remains unclear. HYPOTHESIS: Corylin inhibits the progression of NSCLC cells. METHODS: A lentivector NF-κB luciferase reporter was constructed by molecular cloning. Corylin was screened and identified as an NF-κB pathway inhibitor by luciferase reporter assay. Corylin inhibited the expression of NF-κB downstream genes, which was detected by qRT-PCR. The effect of corylin on NSCLC cells was detected by colony formation assay, cell apoptosis, cell proliferation, in vitro invasion, and cell scratch assay. Corylin inhibited p65 nuclear translocation and was detected by molecular docking, immunofluorescence assay, and Western blot analysis. RESULTS: We constructed a lentiviral expression vector, containing an NF-κB luciferase reporter and established a stable A549 cell line for its expression. Using this cell line, corylin was screened and identified as an NF-κB pathway inhibitor. It was found that corylin inhibited the expression of NF-κB downstream genes and inhibited the proliferation and migration of NSCLC cells. Meanwhile, it was also found that corylin significantly reversed the increased proliferation of NSCLC cell lines induced by p65 overexpression. Molecular docking analysis showed that corylin could bind to p65 by hydrogen bonding. Further study showed that corylin inhibited the NF-κB signaling pathway by blocking p65 nuclear translocation. CONCLUSIONS: Our study screened and identified corylin as an NF-κB inhibitor and elucidated the molecular mechanism by which corylin inhibits the growth of NSCLC cells. The present study provides a novel strategy for improving the prognosis and treatment of NSCLC patients.

Pharmacological Activity of Quercetin: An Updated Review.

Quercetin, a natural flavonoid compound with a widespread occurrence throughout the plant kingdom, exhibits a variety of pharmacological activities. Because of the wide spectrum of health-promoting effects, quercetin has attracted much attention of dietitians and medicinal chemists. An updated review of the literature on quercetin was performed using PubMed, Embase, and Science Direct databases. This article presents an overview of recent developments in pharmacological activities of quercetin including anti-SARS-CoV-2, antioxidant, anticancer, antiaging, antiviral, and anti-inflammatory activities as well as the mechanism of actions involved. The biological activities of quercetin were evaluated both in vitro and in vivo, involving a number of cell lines and animal models, but metabolic mechanisms of quercetin in the human body are not clear. Therefore, further large sample clinical studies are needed to determine the appropriate dosage and form of quercetin for the treatment of the disease.

Adipose-derived mesenchymal stem cells with hypoxic preconditioning improve tenogenic differentiation.

BACKGROUND: Adipose-derived mesenchymal stem cells (ADSCs), as seed cells for tendon tissue engineering, are promising for tendon repair and regeneration. But for ADSCs, diverse oxygen tensions have different stimulatory effects. To explore this issue, we investigated the tenogenic differentiation capability of ADSCs under hypoxia condition (5% O2) and the possible signaling pathways correspondingly. The effects of different oxygen tensions on proliferation, migration, and tenogenic differentiation potential of ADSCs were investigated. METHODS: P4 ADSCs were divided into a hypoxic group and a normoxic group. The hypoxic group was incubated under a reduced O2 pressure (5% O2, 5% CO2, balanced N2). The normoxic group was cultured in 21% O2. Two groups were compared: HIF-1α inhibitor (2-MeOE2) in normoxic culturing conditions and hypoxic culturing conditions. Hypoxia-inducible factor-1α (HIF-1α) and VEGF were measured using RT-qPCR. Specific HIF-1α inhibitor 2-methoxyestradiol (2-MeOE2) was applied to investigate whether HIF-1α involved in ADSCs tenogenesis under hypoxia. RESULTS: Hypoxia significantly reduced proliferation and migration of ADSCs. Continuous treatment of ADSCs at 5% O2 resulted in a remarkable decrease in HIF-1α expression in comparison with 20% O2. Additionally, ADSCs of hypoxia preconditioning exhibited higher mRNA expression levels of the related key tenogenic makers and VEGF than normoxia via RT-qPCR measurement (p ˂ 0.05). Furthermore, the effects of hypoxia on tenogenic differentiation of ADSCs were inhibited by 2-MeOE2. Hypoxia can also stimulate VEGF production in ADSCs. CONCLUSIONS: Our findings demonstrate that hypoxia preconditioning attenuates the proliferation and migration ability of ADSCs, but has positive impact on tenogenic differentiation through HIF-1α signaling pathway.

Effects of hypoxia on Achilles tendon repair using adipose tissue-derived mesenchymal stem cells seeded small intestinal submucosa.

BACKGROUND: The study was performed to evaluate the feasibility of utilizing small intestinal submucosa (SIS) scaffolds seeded with adipose-derived mesenchymal stem cells (ADMSCs) for engineered tendon repairing rat Achilles tendon defects and to compare the effects of preconditioning treatments (hypoxic vs. normoxic) on the tendon healing. METHODS: Fifty SD rats were randomized into five groups. Group A received sham operation (blank control). In other groups, the Achilles tendon was resected and filled with the original tendon (Group B, autograft), cell-free SIS (Group C), or SIS seeded with ADMSCs preconditioned under normoxic conditions (Group D) or hypoxic conditions (Group E). Samples were collected 4 weeks after operation and analyzed by histology, immunohistochemistry, and tensile testing. RESULTS: Histologically, compared with Groups C and D, Group E showed a significant improvement in extracellular matrix production and a higher compactness of collagen fibers. Group E also exhibited a significantly higher peak tensile load than Groups D and C. Additionally, Group D had a significantly higher peak load than Group C. Immunohistochemically, Group E exhibited a significantly higher percentage of MKX + cells than Group D. The proportion of ADMSCs simultaneously positive for both MKX and CM-Dil observed from Group E was also greater than that in Group D. CONCLUSIONS: In this animal model, the engineered tendon grafts created by seeding ADMSCs on SIS were superior to cell-free SIS. The hypoxic precondition further improved the expression of tendon-related genes in the seeded cells and increased the rupture load after grafting in the Achilles tendon defects.

Acute fibrinous and organizing pneumonia: A case report.

RATIONALE: Acute fibrinous and organizing pneumonia (AFOP) is an uncommon type of acute lung injury associated with infection, connective tissue disorders, drug exposure, and hematologic malignancies. PATIENT CONCERNS: A 53-year-old female presented with intermittent fever, chills, and dry cough since 10 days. Chest computed tomography scan showed multiple bilateral patchy infiltrates. PPD skin test was positive but tuberculosis antibody test and T-SPOT were negative. DIAGNOSES: Histologic examination revealed massive fibrinous exudation with organization within alveolar spaces and scattered neutrophilic infiltrates, which was consistent with AFOP. INTERVENTIONS: This patient was treated with prednisolone therapy. OUTCOMES: Chest radiograph improvement and symptom improvement, including fever and respiratory symptoms, was observed after 2 week of oral prednisolone treatment. After 9-month of treatment, the patient was asymptomatic with stable disease and improved quality of life. LESSONS: AFOP has unique pathologic manifestations; however, the condition is liable to be misdiagnosed as community-acquired pneumonia ortuberculosis. Antibiotics are ineffective, while some patients show good response to glucocorticoid therapy.

Relationship between HOX gene and pediatric congenital clubfoot.

The relationship between transcription factor homeobox gene (HOX gene) and pediatric congenital clubfoot (CCF) was studied. The CCF group comprised 35 cases of children, and the control group compised 34 cases of children without congenital malformation. The levels of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) in the serum of the control and CCF groups were measured using iNOS and NO kits. Interleukin-1ß (IL-1ß), IL-6 and tumor necrosis factor-α (TNF-α) related to inflammation in the tissues of both groups were detected by reverse transcription-polymerase chain reaction (RT-PCR). Fatty acid synthase (Fas), Fas ligand (FasL) and Bcl-2-associated X (Bax) related to apoptosis as well as the expression of HOX mRNA, the expression of HOX in the control and CCF groups was detected by western blot analysis, and the differential expression of HOX in the control and CCF groups was statistically analyzed. Results of the kit detection showed that the expression of iNOS and NO in the CCF group were significantly higher than those in the control group, indicating that severe oxidative damage occurred in the CCF group. The results of detecting inflammatory factors and apoptosis by RT-PCR showed that the expression of IL-1ß, IL-6, TNF-α, Fas, FasL and Bax mRNA in the CCF group was significantly higher than that in the control group, indicating pathogenesis of CCF was related to inflammation and apoptosis. RT-PCR and western blot analysis revealed HOX was highly expressed in the tissues of CCF, and the expression quantity was significantly stronger than that in the control group. The result of analysis of variance showed that the expression differences of HOX in normal and CCF tissues were statistically significant (P<0.01). Abnormal expression of HOX was closely related to the occurrence and development of CCF, indicating that HOX has important research value in CCF and this functional mechanism is related to oxidative damage, inflammation and apoptosis. Expression of HOX therefore shows promise as an indicator of CCF diagnosis and treatment.

Effects of NOB1 on the pathogenesis of osteosarcoma and its expression on the chemosensitivity to cisplatin.

The effects of NIN1/RPN12 binding protein 1 homolog (NOB1) on the pathogenesis of osteosarcoma and its expression on the chemosensitivity to cisplatin were investigated. Seventy-four patients with osteosarcoma who received surgical resection in The Affiliated Hospital of Southwest Medical University (Sichuan, China) from September 2013 to September 2016 were enrolled in this study. The expression of NOB1 in cancer and cancer-adjacent tissues of patients was detected by reverse transcription-polymerase chain reaction, and the relationship between NOB1 expression and the pathogenesis of osteosarcoma was analyzed. The expression of NOB1 in osteosarcoma MG-63 cells was interfered with using small interfering ribonucleic acid (siRNA). Western blotting was used to detect the transfection efficiency and changes in apoptosis indicators. Cell Counting Kit-8 (CCK-8) assay was used to examine changes in the sensitivity of cells to cisplatin. The effect of NOB1 knockout on cell apoptosis was examined by flow cytometry. In patients with osteosarcoma, the level of NOB1 mRNA in cancer tissues was significantly higher than that in cancer-adjacent tissues (p<0.05), and the expression of NOB1 was correlated with Ennecking staging and tumor size (p<0.05). The expression level of the apoptotic indicator caspase-3 was activated after siRNA interfered with NOB1 expression, thus reducing the expression level of anti-apoptotic indicator B-cell lymphoma 2. CCK-8 results showed that the downregulation of NOB1 increased the sensitivity of MG-63 cells to cisplatin (p<0.05). In addition, flow cytometry showed that the downregulation of NOB1 significantly promoted the apoptosis of MG-63 cells. NOB1 is significantly upregulated in patients with osteosarcoma, thus reducing the curative effect of cisplatin chemotherapy, which indicates that the prognosis is poor.

[Effectiveness comparison of titanium elastic nail and locking compression plate fixation in treating femoral subtrochanteric fractures in older children].

Objective: To compare the effectiveness of titanium elastic nail (TEN) and locking compression plate fixation in treating femoral subtrochanteric fracture in older children. Methods: Between April 2015 and September 2016, 35 older children (aged 7-13 years) with femoral subtrochanteric fractures were treated, the clinical data were retrospectively reviewed. TEN fixation was used in 19 cases (group A) and locking compression plate fixation in 16 cases (group B). There was no significant difference in age, gender, sides, fracture causes, type of fracture, and time from injury to operation between 2 groups ( P>0.05). The fluoroscopy times, operation time, intraoperative blood loss, fracture healing time were recorded and compared between 2 groups. The limb function was evaluated according to the Sanders scores and Flynn et al. outcome score. Results: All the patients were followed up 6-24 months (mean, 11.46 months). The operation time, intraoperative blood loss, and fracture healing time of group A were significantly less than those of group B, but the fluoroscopy times of group A was significantly more than that of group B ( P<0.05). All the fractures were healed, no breakage of screw, infection of deep tissue, nerve injury, osteonecrosis of the femoral head, or other complication occurred. At last follow-up, according to the Sanders scores, the results were excellent in 14 cases, good in 4 cases, and fair in 1 case in group A with an excellent and good rate of 94.74%; the results were excellent in 12 cases, good in 3 cases, and fair in 1 case in group B with an excellent and good rate of 93.75%; showing no significant difference between 2 groups ( χ2=0.400, P=0.980). According to the Flynn et al. outcome score, the results were excellent in 13 cases, good in 5 cases, and fair in 1 case in group A with an excellent and good rate of 94.74%; the results were excellent in 11 cases, good in 3 cases, and fair in 2 cases in group B with an excellent and good rate of 87.50%; showing no significant difference between 2 groups ( χ2=0.748, P=0.688). Conclusion: Both TEN and locking compression plate have satisfactory outcomes for treating pediatric femoral subtrochanteric fractures. TEN method has minimally trauma, security, and faster fracture healing when compared with locking compression plate.

Cigarette smoking among Chinese PLWHA: An exploration of changes in smoking after being tested HIV positive.

Prevention and cessation of Tobacco use among persons living with HIV/AIDS (PLWHA) represents a significant challenge for HIV/AIDS patient care in China and across the globe. Awareness of HIV-positive status may alter the likelihood for PLWHA smokers to change their smoking habit. In this study, we tested the risk enhancement and risk reduction hypotheses by assessing changes in cigarette smoking behavior among PLWHA after they received the positive results of their HIV tests. Cross-sectional survey data collected from a random sample of 2973 PLWHA in care in Guangxi, China were analyzed. Changes in cigarette smoking after receiving the HIV-positive test results, as well as the current levels of cigarette smoking were measured. Among the total participants, 1529 (51.7%) were self-identified as cigarette smokers, of whom 436 (28.9%) reduced smoking and 286 (19.0%) quit after receiving their HIV-positive test results. Among the quitters, 210 (73.9%) remained abstinent for a median duration of two years. There were also 124 (8.2%) who increased cigarette smoking. Older age, female gender, more education, and receiving antiretroviral therapy were associated with quitting. In conclusion, our study findings support the risk reduction and risk enhancement hypotheses. A large proportion of smoking PLWHA reduced or quit smoking, while a small proportion increased smoking. Findings of this study suggest that the timing when a person receives his or her HIV-positive test result may be an ideal opportunity for care providers to deliver tobacco cessation interventions. Longitudinal studies are indicated to verify the findings of this study and to support smoking cessation intervention among PLWHA in the future.

[EFFECTIVENESS COMPARISON BETWEEN OPEN AND CLOSED REDUCTION USING ELASTIC STABLE INTRAMEDULLARY NAILING FOR PEDIATRIC SUBTROCHANTERIC FRACTURES].

OBJECTIVE: To compare the effectiveness between open reduction and closed reduction of pediatric subtrochanteric fractures using elastic stable intramedullary nailing (ESIN). METHODS: Between January 2010 and January 2014, 22 children with subtrochanteric fractures were treated by ESIN internal fixation combined with hip cast fixation, and the clinical data were retrospectively reviewed. Closed reduction was used in 12 cases (group A) and mini-open reduction in 10 cases (group B). There was no significant difference in gender, age, sides, fracture causes, type of fracture, complications, and time from injury to operation between 2 groups (P > 0.05). The operation time, intraoperative blood loss, fracture healing time, and complications were recorded and compared between 2 groups, the limb function was evaluated according to the Flynn et al. outcome score. RESULTS: The intraoperative blood loss of group A was significantly less than that of group B, but the operation time of group A was significantly longer than that of group B (P < 0.05). All the patients were followed up 12-36 months (mean, 14.9 months). There was no major complications in the other patients of both groups except 1 patient having delayed wound healing in group B. There was no significant difference in fracture healing time between 2 groups (t = -1.006, P = 0.327). Inverted angle of 10° and shortened limb of 1.8 cm were observed in 1 case of group A, and sagittal plane angle of 15° and shortened limb of 2 cm in 1 case of group B. There was no abnormal walking and function of hip and knee activity at last follow-up. According to the Flynn et al. outcome score, the results were excellent in 8 cases and good in 4 cases in group A, and were excellent in 6 cases and good in 4 cases in group B, showing no significant difference between 2 groups (χ2 = 0.041, P = 0.956). CONCLUSION: Both closed and open fracture reduction using ESIN have satisfactory outcomes for treating pediatric subtrochanteric fractures. A mini-open reduction should be selected intraoperatively if closed reduction proves to be difficult.

[Elastic stable intramedullary nailing for treatment of benign lesions of humerus complicating by pathological fracture in children].

OBJECTIVE: To explore the effectiveness and advantages of elastic stable intramedullary nail (ESIN) combined with curettage and graft for the treatment of benign lesions of humerus complicating by pathological fracture in children. METHODS: ESIN internal fixation combined with curettage and graft was used to treat benign lesions of the humerus complicating by pathological fracture in 11 children patients between January 2007 and January 2011. Of 11 patients, 7 were boy and 4 were girl, aged from 5 to 14 years (mean, 9.4 years). The disease duration ranged from 2 to 14 days (mean, 6 days). All fractures were closed fracture, which locations were the proximal humerus in 6 cases, the humeral shaft in 4 cases, and the distal humerus in 1 case; benign lesions of the humerus included aneurysmal bone cyst in 1 case, simple bone cyst in 7 cases, and fibrous dysplasia in 3 cases. Based on imaging studies, preoperative diagnosis was almost clear. The time from hospitalization to operation was 3-5 days. RESULTS: Healing of incision by first intention was obtained in all cases, with no infection. The mean follow-up was 25.6 months (range, 12-36 months). All patients achieved pain relief at 6 weeks postoperatively and fractures healed completely at 3 to 4 months after operation (mean, 3.3 months). No recurrence or re-fracture was observed during follow-up. The ESIN was removed at 10-14 months after operation (mean, 12.5 months). The lesion disappeared completely in 8 cases and partially in 3 cases. No pain of affected limb or motion limitation of shoulder and elbows was observed. One patient had limb shortening of 2 cm at last follow-up, but he had no function problem. According to Neer shoulder and Mayo elbow function scores, the results were excellent in 11 cases. CONCLUSION: It is a good method to treat benign lesions of the humerus complicating by pathological fracture in children to use ESIN internal fixation combined with curettage and graft. After only a single operation intervention, it can provide early mechanical stability and rapid fracture healing and allow early rehabilitation exercise.