Objective To investigate the effect of endothelial cells in the perivascular niche on the stem cell-like characteristics of osteosarcoma cells and primarily explore the possible molecular mechanism.Methods A co-culture model was established in vitro using SV40T-human umbilical vein endothelial cells(HUVEC-T1)and osteosarcoma stem cells(OSCs)derived from the human osteosarcoma cell line 143B.Thus,there were 2 groups of cells,OSCs and co-cultured OSCs.The self-renewal capacity of OSCs between the 2 groups was assessed using a limiting dilution forming sphere assay.Flow cytometry and Western blotting were used to detect the expression of stem cell marker CD133 and stemness transcription factors SOX2 and NANOG.Fourteen female nude mice(4～6 weeks old,weighing 18～20 g)were randomly and equally divided into 2 groups of subcutaneous xenograft models:the control group(OSCs suspension)and the experimental group(OSCs+HUVEC-T1 mixed suspension).The tumor volume and mass were compared between the 2 xenograft groups.Immunofluorescence(IF)staining was used to verify the spatial proximity between endothelial cells and OSCs in vivo,while immunohistochemistry was employed to compare microvessel density(MVD)and CD133 expression level between the 2 groups.RNA-seq was performed to identify potential signaling pathways of endothelial cells affecting the stemness of OSCs.PCR and Western blotting were applied to confirm the RNA-seq findings.Exogenous protein treatment,IF staining,Western blotting and sphere formation assay were utilized to preliminarily validate the role of the identified pathway in regulating the stemness phenotype of OSCs.Results The in vitro co-culture model of HUVEC-T1 and OSCs was successfully established.Compared with the control group,the co-culture group exhibited significantly enhanced self-renewal ability of OSCs,laeger proportion of the stemness marker CD133+[(8.20±1.64)％vs(4.32±1.34)％,P<0.05],enhanced expression of CD117,SOX2 and NANOG(P<0.05),along with more sphere formation(P<0.05)and elevated SOX2/NANOG protein levels.The xenograft mice from the experimental group showed larger tumor volume(643.10±413.50 vs 247.90±93.66 mm3,P<0.05)and heavier tumor weight(0.52±0.27 vs 0.24±0.10 g,P<0.05)when compared the control group,correspondingly showing increased MVD(22.57±11.84 vs 11.43±5.38,P<0.05)and elevated CD133 expression(P<0.05).IF staining confirmed the adjacency of CD31-labeled endothelial cells and CD133-labeled OSCs in vivo.RNA-seq and functional experiments demonstrated that CXCR4 was highly expressed in co-cultured OSCs,CXCL12 was highly expressed in co-cultured endothelial cells,and exogenous CXCL12 promoted the sphere formation and expression levels of SOX2 and NANOG of OSCs(P<0.05).Conclusion Endothelial cells within the perivascular niche may promote the stemness phenotype of osteosarcoma cells via the CXCL12/CXCR4 pathway.

Objective To explore the molecular mechanism of Atractylodes macrocephala in the treatment of Ulcerative colitis(UC)based on network pharmacology,and verify it with animal experiments.Methods The active components of Atractylodes macrocephala was screened from the TCMSP database,the TCM-ID database,and in combination with relevant references,and the corresponding targets were obtained through Swiss database.The relevant targets of UC were obtained from GeneCards database,construct the"drug-component-target-disease"network diagram and"pathway-active ingredient-target"network diagram and draw PPI network diagram;GO function enrichment analysis and KEGG signal pathway annotation analysis were carried out.Autodock software is used for molecular docking of active components and targets.Then,the experimental validation of the network pharmacology prediction was carried out.The mouse UC model was induced by dextran sodium sulfate(DSS).The pathological changes of the colon tissue,the number of goblet cells,and the positive expression of inflammatory factorswere detected by HE staining,AB-PAS staining and immunohistochemistry in colon tissue of UC mice.Results The results have shown 30 active ingredients including atractylolactone I,II and III were screened,and 591 corresponding targets were obtained,of which the key target was IL-1β、TNF-α and so on.Molecular docking show that the core components had good binding affinity with the key targets.And the results of animal experiments showed that the alcohol extract of Atractylodes macrocephala could significantly increase the colon length,reduce the DAI score,improve the pathological changes of colon tissue of UC mice,increase the number of goblet cells,and inhibit the expression of IL-1β,TNF-α in colon tissue.Conclusion This study indicated that Atractylodes macrocephala could regulate the release of inflammatory factors through multiple components,multi-target and multi-channel,which could inhibit inflammatory reaction and play a role in improving UC.

Objective:To discuss how to make the surgical strategy for tibial tubercle fracture associated with bicondylar tibial plateau fracture.Methods:Data of thirty-five patients of tibial tubercle fractures associated with bicondylar tibial plateau fractures who were treated from October 2014 to May 2018 were retrospectively analyzed. There were 26 males and 9 females with an average age of 37.6 years (range, 21-68 years). According to Schatzker classification in tibial plateau fracture, 16 cases were type V and 19 cases were type VI. According to the integrity of tibial tubercle fracture and cortical bone of the proximal tibia in bicondylar tibial plateau fracture, they were divided into four types: type A, tibial tubercle fracture fragment and cortical bone of the proximal tibia are both complete; type B, tibial tubercle fracture fragment is complete but cortical bone of the proximal tibia is comminuted; type C, tibial tubercle fracture fragment is comminuted but cortical bone of the proximal tibia is complete; type D, both of them are comminuted. The surgical approaches and fixation methods of all the tibial tubercle fractures were according to the four different types. There were 22 cases with type A and B that were treated via an anterolateral and a medial incision, 13 cases with type C and D were treated via an anterior midline and a medial incision. There were 4 cases belonging to type A fixed with lag screws singly, 18 cases with type B fixed with 1/4 tubular plates, 7 cases with type C and 6 cases with type D fixed by 1/4 tubular plates combined with lag screws.Results:Thirty-five patients were followed up for 16.8 months (range, 12-24 months). All fractures healed with an average time of 4.7 months (range, 3-6 months). Loss of reduction didn’t occur in 34 cases except one. According to Rasmussen radiographic evaluation, the average score was 14.1 (range, 10-18) and clinical outcomes were rated with excellent in 11 cases, good in 19, fair in 5. The excellent and good rate was 85.7% (30/35) . The mean Hospital for Special Surgery (HSS) scores of all cases were 86.8 (range, 64-98) and the functional scores were excellent in 22 cases, good in 10 cases and fair in 3 cases with the excellent and good rate of 91.4% (32/35) . Surgical complications included fat liquefaction in 2 cases, superficial wound infection in 1, loosening of implant in 1and traumatic arthritis in 1.Conclusion:This kind of tibial tubercle fracture associated with bicondylar tibial plateau fracture is rare and special. Therefore, the preoperative plan should be made by considering the morphological features of the tibial tubercle fragments and the cortical bone of the proximal tibia. The middle longitude approach is the best way to expose tibial tubercle fragments which should be fixed with 1/4 tubularplate and/or lag screws.

OBJECTIVE: To exploit material decomposition analysis in dual-energy spectral computed tomography (CT) to assess the blood supply status of the ground-glass opacity (GGO) in lungs. MATERIALS AND METHODS: This retrospective study included 48 patients with lung adenocarcinoma, who underwent a contrast-enhanced dual-energy spectral CT scan before treatment (53 GGOs in total). The iodine concentration (IC) and water content (WC) of the GGO, the contralateral and ipsilateral normal lung tissues were measured in the arterial phase (AP) and their differences were analyzed. IC, normalized IC (NIC), and WC values were compared between the pure ground-glass opacity (pGGO) and the mixed ground-glass opacity (mGGO), and between the group of preinvasive lesions and the minimally invasive adenocarcinoma (MIA) and invasive adenocarcinoma (IA) groups. RESULTS: The values of pGGO (IC = 20.9 ± 6.2 mg/mL and WC = 345.1 ± 87.1 mg/mL) and mGGO (IC = 23.8 ± 8.3 mg/mL and WC = 606.8 ± 124.5 mg/mL) in the AP were significantly higher than those of the contralateral normal lung tissues (IC = 15.0 ± 4.9 mg/mL and WC = 156.4 ± 36.8 mg/mL; IC = 16.2 ± 5.7 mg/mL and WC = 169.4 ± 41.0 mg/mL) and ipsilateral normal lung tissues (IC = 15.1 ± 6.2 mg/mL and WC = 156.3 ± 38.8 mg/mL; IC = 15.9 ± 6.0 mg/mL and WC = 174.7 ± 39.2 mg/mL; all p < 0.001). After normalizing the data according to the values of the artery, pGGO (NIC = 0.1 and WC = 345.1 ± 87.1 mg/mL) and mGGO (NIC = 0.2 and WC = 606.8 ± 124.5 mg/mL) were statistically different (p = 0.049 and p < 0.001, respectively), but not for the IC value (p = 0.161). The WC values of the group with preinvasive lesions and MIA (345.4 ± 96.1 mg/mL) and IA (550.1 ± 158.2 mg/mL) were statistically different (p < 0.001). CONCLUSION: Using dual-energy spectral CT and material decomposition analysis, the IC in GGO can be quantitatively measured which can be an indicator of the blood supply status in the GGO.
Adenocarcinoma ; Arteries ; Humans ; Iodine ; Lung ; Multidetector Computed Tomography ; Retrospective Studies ; Tomography, X-Ray Computed ; Water

Objective To investigate the clinical efficacy of a period of joint fusion and plate internal fixation for four parts calcaneal fractures.Methods The clinical data of 53 patients with four parts calcaneal fractures from January 2009 to January 2013 were analyzed retrospectively.Twenty-two patients were performed a period of joint fusion (joint fusion group),31 patients were performed plate internal fixation (plate internal fixation group).The Bohler angle and Gissane corner before and after operation,operation time and bleeding,the Maryland foot function score between two groups were compared.Results Compared with before operation,the Bohler angle and Gissane corner in plate internal fixation group and joint fusion group were improved significantly after operation (34.6° ± 4.2° vs.5.7° ± 2.4°,125.4° ± 8.2° vs.87.2° ± 6.8°,32.8° ± 3.9° vs.5.4° ± 2.7°,127.6° ± 7.8° vs.86.9° ± 6.7°),and there were significant differences (P ＜0.05),but the Bohler angle and Gissane corner before and after operation between two groups had no significant difference (P ＞ 0.05).The operation time and bleeding were (52.1 ± 5.1) min,(50.0 ± 10.2) ml in plate internal fixation group and (70.3 ± 5.2) min,(105.0 ± 20.5) ml in joint fusion group,and there were significant differences between two groups (P ＜ 0.05).The fineness rate of Maryland foot function at 6 months follow-up between two groups had no significant difference (P ＞ 0.05).Conclusions The plate internal fixation and joint fusion is therapeutic equivalence in treatment of four parts calcaneal fractures.Due to the small surgical trauma and less bleeding of plate internal fixation,it may be preferred in the treatment of four parts calcaneal fractures.