Objective To investigate the mechanism of astragalus polysaccharide(APS)in inhibiting the proliferation and metastasis of osteosarcoma cells.Methods Cell counting kit-8(CCK8)assay was used to detect the inhibitory effect of APS on the proliferation of osteosarcoma cell lines U2OS,HOS,MG63 and osteoblast cell line hFOB1.19,the cell line with the most significant inhibitory effect of APS on osteosarcoma cells was selected for subsequent experiments.Osteosarcoma cells were divided into control group(NC group),APS group,APS+sh-SP1 co treatment group with transfected SP1 knockdown plasmid(APS+sh-SP1),and APS+oe SP1 co treatment group with transfected SP1 overexpression plasmid(APS+oe SP1 group).Western blotting was used to detect the expression of SP1 protein in each group.CCK8 assay was used to detect the proliferation ability of each group.Transwell assay was used to detect the metastatic ability of cells in each group.TOP/FOP Flash assay was used to detect the activity of Wnt/β-catenin signaling pathway.The protein expressions of Wnt3a,β-catenin,CyclinD1,cMYC,matrix metalloproteinase 2(MMP2)and Snail were detected by western blotting.Result After 48h of APS treatment,the cell proliferation inhibition rates of osteosarcoma cells U2OS,HOS,MG63 and osteoblast cells hFOB1.19 were 62.93%±4.79%,20.66%±1.10%,39.31%±3.20%and 5.97%±0.72%,respectively.Compared with osteoblast hFOB1.19,APS significantly inhibited the proliferation of osteosarcoma cells,and the difference was statistically significant(F=208.400,P<0.001),and the inhibitory effect on osteosarcoma U2OS cells was the most significant(t=20.380,P<0.001).Compared with NC group,SP1 protein expression,cell proliferation ability,number of transmembrane cells,Wnt/β-catenin signaling pathway activity in cells,Wnt/β-catenin signaling pathway key proteins Wnt3a,β-catenin,downstream proliferation-related protein CyclinD1,cMYC,and downstream metastasis related proteins MMP2 and Snail in the APS group were decreased,and the differences were statistically significant(t=9.740～90.780,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+sh-SP1 group were further decreased,and the differences were statistically significant(t=3.032～12.940,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+oe-SP1 group were increased,and the differences were statistically significant(t=3.350～22.450,all P<0.05).Conclusion APS inhibits the proliferation and metastasis of osteosarcoma cells by targeting SP1/Wnt/β-catenin signaling axis.

Objective To investigate the mechanism of astragalus polysaccharide(APS)in inhibiting the proliferation and metastasis of osteosarcoma cells.Methods Cell counting kit-8(CCK8)assay was used to detect the inhibitory effect of APS on the proliferation of osteosarcoma cell lines U2OS,HOS,MG63 and osteoblast cell line hFOB1.19,the cell line with the most significant inhibitory effect of APS on osteosarcoma cells was selected for subsequent experiments.Osteosarcoma cells were divided into control group(NC group),APS group,APS+sh-SP1 co treatment group with transfected SP1 knockdown plasmid(APS+sh-SP1),and APS+oe SP1 co treatment group with transfected SP1 overexpression plasmid(APS+oe SP1 group).Western blotting was used to detect the expression of SP1 protein in each group.CCK8 assay was used to detect the proliferation ability of each group.Transwell assay was used to detect the metastatic ability of cells in each group.TOP/FOP Flash assay was used to detect the activity of Wnt/β-catenin signaling pathway.The protein expressions of Wnt3a,β-catenin,CyclinD1,cMYC,matrix metalloproteinase 2(MMP2)and Snail were detected by western blotting.Result After 48h of APS treatment,the cell proliferation inhibition rates of osteosarcoma cells U2OS,HOS,MG63 and osteoblast cells hFOB1.19 were 62.93%±4.79%,20.66%±1.10%,39.31%±3.20%and 5.97%±0.72%,respectively.Compared with osteoblast hFOB1.19,APS significantly inhibited the proliferation of osteosarcoma cells,and the difference was statistically significant(F=208.400,P<0.001),and the inhibitory effect on osteosarcoma U2OS cells was the most significant(t=20.380,P<0.001).Compared with NC group,SP1 protein expression,cell proliferation ability,number of transmembrane cells,Wnt/β-catenin signaling pathway activity in cells,Wnt/β-catenin signaling pathway key proteins Wnt3a,β-catenin,downstream proliferation-related protein CyclinD1,cMYC,and downstream metastasis related proteins MMP2 and Snail in the APS group were decreased,and the differences were statistically significant(t=9.740～90.780,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+sh-SP1 group were further decreased,and the differences were statistically significant(t=3.032～12.940,all P<0.05).Compared with the APS group,the expression of SP1 protein,cell proliferation ability,the number of transmembrane cells,the activity of Wnt/β-catenin signaling pathway and the expression of Wnt/β-catenin signaling pathway related proteins in the APS+oe-SP1 group were increased,and the differences were statistically significant(t=3.350～22.450,all P<0.05).Conclusion APS inhibits the proliferation and metastasis of osteosarcoma cells by targeting SP1/Wnt/β-catenin signaling axis.

Objective:To determine the pathogens causing viral diarrhea in children under 5 years of age and the epidemiological patterns in Chengdu from 2021 to 2024.Methods:Fecal samples were collected from hospitalized children under 5 years of age with diarrhea in a sentinel surveillance hospital for viral diarrhea in Chengdu from 2021 to 2024. Norovirus（NoV），sapovirus（SaV），astrovirus（AstV），and enteric adenovirus（EAdV）were detected by real-time PCR or real-time RT-PCR. Group A rotavirus（RVA）was detected by enzyme-linked immunosorbent assay. The results were analyzed statistically by using SPSS 20.0 software.Results:RVA，NoV，SaV，AstV，and EAdV were detected in 329 fecal samples from 719 hospitalized children with diarrhea under 5 years of age. The total detection rate was 48.8%（329/719）. The detection rate of RVA，NoV，SaV，AstV，and EAdV was 17.0%（122/719），20.9%（150/719），4.0%（29/719），2.8%（20/719），8.6%（62/719），separately. The detection rate of mixed infection with multiple pathogens was 7.1%（51/719），among which RVA combined with NoV infection was the most common（2.2%，16/719）. Except for EAdV（ χ2=14.368， P=0.002），there was no significant difference in the detection rates of RVA，NoV，SaV，and AstV in different years. Except for SaV and EAdV，the detection rates of RVA（ χ2=11.106， P<0.001），NoV（ χ2=65.164， P<0.001）and AstV（ χ2=19.612， P<0.001）were statistically significant in different quarters. There was no statistically significant difference in the detection rates of the five pathogens between males and females. Except for NoV，there were statistically significant differences in the detection rates of RVA（ χ2=32.209， P<0.001），SaV（ χ2=13.983， P=0.002），AstV（ χ2=9.56， P=0.015）and EAdV，（ χ2=8.992， P=0.029）in children of different months of age. Conclusion:RVA and NoV are the main pathogens causing viral diarrhea in children under 5 years of age in Chengdu. The peak infection period of viral diarrhea tends to occur in winter and spring and occurs most frequently in 12-24 months old and 24-59 months old children.

Objective:To determine the pathogens causing viral diarrhea in children under 5 years of age and the epidemiological patterns in Chengdu from 2021 to 2024.Methods:Fecal samples were collected from hospitalized children under 5 years of age with diarrhea in a sentinel surveillance hospital for viral diarrhea in Chengdu from 2021 to 2024. Norovirus（NoV），sapovirus（SaV），astrovirus（AstV），and enteric adenovirus（EAdV）were detected by real-time PCR or real-time RT-PCR. Group A rotavirus（RVA）was detected by enzyme-linked immunosorbent assay. The results were analyzed statistically by using SPSS 20.0 software.Results:RVA，NoV，SaV，AstV，and EAdV were detected in 329 fecal samples from 719 hospitalized children with diarrhea under 5 years of age. The total detection rate was 48.8%（329/719）. The detection rate of RVA，NoV，SaV，AstV，and EAdV was 17.0%（122/719），20.9%（150/719），4.0%（29/719），2.8%（20/719），8.6%（62/719），separately. The detection rate of mixed infection with multiple pathogens was 7.1%（51/719），among which RVA combined with NoV infection was the most common（2.2%，16/719）. Except for EAdV（ χ2=14.368， P=0.002），there was no significant difference in the detection rates of RVA，NoV，SaV，and AstV in different years. Except for SaV and EAdV，the detection rates of RVA（ χ2=11.106， P<0.001），NoV（ χ2=65.164， P<0.001）and AstV（ χ2=19.612， P<0.001）were statistically significant in different quarters. There was no statistically significant difference in the detection rates of the five pathogens between males and females. Except for NoV，there were statistically significant differences in the detection rates of RVA（ χ2=32.209， P<0.001），SaV（ χ2=13.983， P=0.002），AstV（ χ2=9.56， P=0.015）and EAdV，（ χ2=8.992， P=0.029）in children of different months of age. Conclusion:RVA and NoV are the main pathogens causing viral diarrhea in children under 5 years of age in Chengdu. The peak infection period of viral diarrhea tends to occur in winter and spring and occurs most frequently in 12-24 months old and 24-59 months old children.

Objective:To explore the evaluation effect of ultrasonography and Pirani score on tarsal deformity, treatment effect and pseudo-correction of congenital clubfoot in infants and young children, and the correlation between the two methods.Methods:This is a retrospective case series study. The clinical data of 26 children (40 feet) with congenital clubfoot who were evaluated by ultrasonography in the Third Affiliated Hospital of Zhengzhou University from January 2020 to January 2023 were retrospectively collected. There were 16 males and 10 females. The age at the first ultrasound examination was ( M(IQR)) 9.0 (18.0) days (range: 1 to 46 days). All patients were treated with Ponseti method by the same physician. The Pirani scores before and after treatment and at the last examination, and the talonavicular angle, calcaneocuboid angle and tibiocalcaneal angle measured by ultrasound were collected, and the treatment and follow-up were recorded. Paired sample t test, repeated measures analysis of variance or Kruskal-Wallis test were used for data comparison, and Spearman correlation analysis was used for correlation analysis. The receiver operating characteristic curve was used to calculate the efficacy of ultrasound in evaluating different Pirani scores. Results:The number of plaster fixation in 26 children was 4.0 (1.0) times (range: 2 to 8 times). The medial talonavicular angle and posterior tibiocalcaneal angle were significantly improved after treatment and at the last follow-up compared with those before treatment, and the differences were statistically significant (all P<0.01). There was no difference in lateral calcaneocuboid angle before and after treatment and at the last follow-up ( F=1.971, P>0.05). Pseudo-correction occurred in 2 cases (2 feet) during the treatment, with an incidence of 5%. Correlation analysis showed that there was a moderate positive correlation between talonavicular angle and Pirani midfoot score ( r=0.480, P<0.01). There was no correlation between calcaneocuboid angle and Pirani midfoot score ( r=0.114, P=0.105). There was a moderate negative correlation between tibial heel angle and Pirani hindfoot score ( r=-0.566, P<0.01). The cut-off point of Pirani midfoot score of 1.5 was 38.78°, the sensitivity was 0.90, the specificity was 0.56, and the area under the curve was 0.75. The cut-off value of angle was 27.51 °, the sensitivity was 0.16, the specificity was 0.92, and the area under the curve was 0.44.The cut-off points of Pirani midfoot score of 3.0 were 45.08°and 9.96°, the sensitivity was 0.94 and 0.91, the specificity was 0.37 and 0.42, and the area under the curve was 0.59 and 0.62, respectively. The cut-off values of Pirani hindfoot score of 2.0 and 3.0 were 167.46° and 160.15°, respectively. The sensitivity was 0.75 and 0.67, the specificity was 0.81 and 0.83, and the area under the curve was 0.78 and 0.71, respectively. Conclusion:Ultrasound can complement with Pirani score, visually and dynamically observe the morphology and position changes of talonavicular joint, calcaneocuboid joint and tibiotalocalcaneal joint, monitor the recovery and pseudo-correction of tarsal bones, and better evaluate the therapeutic effect.

Objective:To explore the evaluation effect of ultrasonography and Pirani score on tarsal deformity, treatment effect and pseudo-correction of congenital clubfoot in infants and young children, and the correlation between the two methods.Methods:This is a retrospective case series study. The clinical data of 26 children (40 feet) with congenital clubfoot who were evaluated by ultrasonography in the Third Affiliated Hospital of Zhengzhou University from January 2020 to January 2023 were retrospectively collected. There were 16 males and 10 females. The age at the first ultrasound examination was ( M(IQR)) 9.0 (18.0) days (range: 1 to 46 days). All patients were treated with Ponseti method by the same physician. The Pirani scores before and after treatment and at the last examination, and the talonavicular angle, calcaneocuboid angle and tibiocalcaneal angle measured by ultrasound were collected, and the treatment and follow-up were recorded. Paired sample t test, repeated measures analysis of variance or Kruskal-Wallis test were used for data comparison, and Spearman correlation analysis was used for correlation analysis. The receiver operating characteristic curve was used to calculate the efficacy of ultrasound in evaluating different Pirani scores. Results:The number of plaster fixation in 26 children was 4.0 (1.0) times (range: 2 to 8 times). The medial talonavicular angle and posterior tibiocalcaneal angle were significantly improved after treatment and at the last follow-up compared with those before treatment, and the differences were statistically significant (all P<0.01). There was no difference in lateral calcaneocuboid angle before and after treatment and at the last follow-up ( F=1.971, P>0.05). Pseudo-correction occurred in 2 cases (2 feet) during the treatment, with an incidence of 5%. Correlation analysis showed that there was a moderate positive correlation between talonavicular angle and Pirani midfoot score ( r=0.480, P<0.01). There was no correlation between calcaneocuboid angle and Pirani midfoot score ( r=0.114, P=0.105). There was a moderate negative correlation between tibial heel angle and Pirani hindfoot score ( r=-0.566, P<0.01). The cut-off point of Pirani midfoot score of 1.5 was 38.78°, the sensitivity was 0.90, the specificity was 0.56, and the area under the curve was 0.75. The cut-off value of angle was 27.51 °, the sensitivity was 0.16, the specificity was 0.92, and the area under the curve was 0.44.The cut-off points of Pirani midfoot score of 3.0 were 45.08°and 9.96°, the sensitivity was 0.94 and 0.91, the specificity was 0.37 and 0.42, and the area under the curve was 0.59 and 0.62, respectively. The cut-off values of Pirani hindfoot score of 2.0 and 3.0 were 167.46° and 160.15°, respectively. The sensitivity was 0.75 and 0.67, the specificity was 0.81 and 0.83, and the area under the curve was 0.78 and 0.71, respectively. Conclusion:Ultrasound can complement with Pirani score, visually and dynamically observe the morphology and position changes of talonavicular joint, calcaneocuboid joint and tibiotalocalcaneal joint, monitor the recovery and pseudo-correction of tarsal bones, and better evaluate the therapeutic effect.

Objective:To investigate the clinicopathological and genetic features of epithelioid and spindle cell rhabdomysarcoma with EWSR1-TFCP2 or FUS-TFCP2 fusion.Methods:The clinical, morphological and immunohistochemical features of 14 cases of epithelioid and spindle cell rhabdomysarcoma with EWSR1-TFCP2 or FUS-TFCP2 fusion diagnosed from January 2019 to December 2022 in the Department of Pathology, Foshan Traditional Chinese Medicine Hospital, Foshan, China were retrospectively analyzed. The cases were all subject to FISH or next generation sequencing for analysis of molecular genetic features. The literature was reviewed.Results:There were 5 males and 9 females, with the age at presentation ranging from 6 to 36 years (mean, 22 years). Tumors occurred in the head and neck (9 cases), pelvic region (2 cases), bladder (one case), right humerus (one case), and the abdominal wall, humerus and pubic at the same time (one case). Presenting symptoms varied by location but often included pain or discomfort. Most of the patients showed aggressive radiographic features with soft tissue extension. The tumors had a median size of 6.6 cm (range, 2-23 cm). The tumors were poorly defined and irregularly shaped. Microscopic examination showed diffuse proliferation of spindle or epithelioid cells. While morphologically high-grade tumors displayed obvious cytological atypia, a high mitotic count and tumor necrosis, low-grade tumors grew in sheets and fascicles composed of spindle, epithelioid cells with moderate or abundant amounts of eosinophilic cytoplasm, without pronounced cytological atypia. The tumor cells expressed Desmin, MyoD1, and Myogenin, as well as ALK, EMA, and CKpan. EWSR1/FUS-TFCP2 gene fusion was detected in 14 cases with next generation sequencing and confirmed by FISH. Six cases had EWSR1-TFCP2 fusions and 8 cases showed FUS-TFCP2 fusions. Follow-up information was available in 13 patients, ranged from 5 to 37 months. At the end of follow-up period, 7 patients died of the disease. Six patients were alive:two cases had local recurrences and metastases, two cases of recurrences, one case of metastasis and one case without recurrences and metastasis.Conclusions:Epithelioid and spindle cell rhabdomysarcomas with EWSR1-TFCP2 or FUS-TFCP2 fusion show a very aggressive clinical course, and more commonly occur in the head and neck. Their genetic hallmark is the presence of EWSR1/FUS-TFCP2 fusions. Familiarity with its clinicopathological characteristics is helpful in avoiding misdiagnoses.

Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.

Objective To investigate the value of a nomogram model constructed from intratumoral and peritumoral magnetic resonance imaging radiomics combined with clinical characteristics in predicting the status of lymphovascular space invasion(LVSI)in cervical cancer.Methods A retrospective analysis was conducted on 178 cervical cancer patients confirmed by postoperative pathology,with 70 cases of LVSI(+)and 108 cases of LVSI(-).The patients were divided into a training set[142 cases,including 54 cases of LVSI(+)and 88 cases of LVSI(-)]and a test set[36 cases,including 16 cases of LVSI(+)and 20 cases of LVSI(-)]at a ratio of 8:2.All underwent magnetic resonance imaging before surgery,and regions of interest were manually delineated layer by layer on the T2WI sequence,with the peritumoral region being uniformly expanded outward.Univariate logistic analysis was performed on clinical factors to select independent factors for cervical cancer LVSI(+).Radiomic features were extracted separately from the intratumoral region,the peritumoral region,and the intratumoral-peritumoral region to construct radiomics models,and the differences between the peritumoral and the intratumoral-peritumoral models were compared.A combined model was established based on the radiomics scores of the optimal intratumoral-peritumoral model and clinical independent predictive factors,and a nomogram was plotted.Receiver operating characteristic curves were used to evaluate the diagnostic performance of each model,and decision curve analysis was used to assess the clinical value of the models.Results The combined model demonstrated the best performance among the various models,with AUC of 0.970 in the training set and 0.803 in the test set.Conclusion Intratumoral and peritumoral magnetic resonance imaging radiomics combined with clinical characteristics can effectively predict LVSI in cervical cancer.

BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice.MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH 2 O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected.Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.