[Effectiveness of minimally invasive treatment of hallux valgus with small incision external articular osteotomy].

Objective: To compare the effectiveness of small incision external articular minimally invasive osteotomy and traditional Chevron osteotomy in the treatment of hallux valgus. Methods: A retrospective analysis was conducted on the clinical data of 58 patients (58 feet) with hallux valgus who were admitted between April 2019 and June 2022 and met the selection criteria. Among them, 28 cases were treated with small incision external articular minimally invasive osteotomy (minimally invasive group), and 30 cases were treated with traditional Chevron osteotomy (traditional group). There was no significant difference in baseline data such as age, gender, disease duration, Mann classification, and preoperative inter metatarsal angle (IMA), hallux valgus angle (HVA), distal metatarsal articular angle (DMAA), forefoot width, tibial sesamoid position (TSP) score, American Orthopaedic Foot and Ankle Society (AOFAS) forefoot score, visual analogue scale (VAS) score, psychological score (SF-12 MCS score) and physiological score (SF-12 PCS score) of short-form 12 health survey scale, and range of motion (ROM) of metatarsophalangeal joint between the two groups ( P>0.05). The incision length, operation time, intraoperative blood loss, intraoperative fluoroscopy frequency, weight-bearing walking time, fracture healing time, and incidence of complications were recorded and compared between the two groups; as well as the changes of imaging indexes at last follow-up, and the clinical function score and ROM of metatarsophalangeal joint before operation, at 6 weeks after operation, and at last follow-up. Results: All patients were followed up 11-31 months, with an average of 22 months. The incision length and intraoperative blood loss in the minimally invasive group were significantly less than those in the traditional group ( P<0.05), and the intraoperative fluoroscopy frequency and operation time in the minimally invasive group were significantly more than those in the traditional group ( P<0.05); but no significant difference was found in weight-bearing walking time and fracture healing time between the two groups ( P>0.05). There was 1 case of skin injury in the minimally invasive group and 3 cases of poor incision healing in the traditional group; all patients had good healing at the osteotomy site, and no complication such as infection, nerve injury, or metatarsal head necrosis occurred. At last follow-up, the imaging indexes of the two groups significantly improved when compared with those before operation ( P<0.05). The changes of DMAA and TSP score in the minimally invasive group were significantly better than those in the traditional group ( P<0.05), and there was no significant difference in the changes of IMA, HVA, and forefoot width between the two groups ( P>0.05). The clinical scores and ROM of metatarsophalangeal joint significantly improved in the two groups at 6 weeks after operation and at last follow-up when compared with preoperative ones ( P<0.05), and the indicators in the minimally invasive group were significantly better than those in the traditional group ( P<0.05). Conclusion: Compared with traditional Chevron osteotomy, small incision external articular minimally invasive osteotomy can effectively improve HVA, IMA, and forefoot width, correct foot deformities, and has less trauma. It can better correct the first metatarsal pronation deformity and restore the anatomical position of the sesamoid bone, resulting in better effectiveness.

[Content of bone morphogenetic protein 2 in demineralized bone matrix prepared from different long bones and study of the osteogenic properties in vitro].

Objective: To measure the concentration of bone morphogenetic protein 2 (BMP-2) in demineralized bone matrix (DBM) prepared from different long bones and to evaluate the osteoinductivity of different DBM on MC3T3-E1 cells. Methods: Different bones from the same cadaver donor were used as the initial materials for making DBM, which were divided into ulna group (uDBM), humerus group (hDBM), tibia group (tDBM), and femur group (fDBM) according to the origins, and boiled DBM (cDBM) was taken as the control group. The proteins of DBM were extracted by guanidine hydrochloride, and the concentrations of BMP-2 were determined by ELISA assay. Then the DBM were co-cultured with MC3T3-E1 cells, the proliferation of MC3T3-E1 cells was observed by cell counting kit 8 (CCK-8) assay. The osteogenic differentiation ability of MC3T3-E1 cells was qualitatively observed by alizarin red, alkaline phosphatase (ALP), and Van Gieson staining, and the osteogenic differentiation ability of MC3T3-E1 cells was quantitatively analyzed by ALP content. Linear regression was used to analyze the effect of BMP-2 concentration in DBM on ALP synthesis. Results: There were significant differences in the concentration of BMP-2 among the DBM groups (P<0.05). The concentrations of BMP-2 in the lower limb long bone were higher than those in the upper limb long bone, and the concentration of BMP-2 in the fDBM group was about 35.5 times that in the uDBM group. CCK-8 assay showed that the cells in each group continued to proliferate within 5 days of co-culture, and the absorbance (A) values at different time points were in the order of cDBM group

Hybrid double-spiral microfluidic chip for RBC-lysis-free enrichment of rare cells from whole blood.

Drug selection and treatment monitoring via minimally invasive liquid biopsy using circulating tumor cells (CTCs) are expected to be realized in the near future. For clinical applications of CTCs, simple, high-throughput, single-step CTC isolation from whole blood without red blood cell (RBC) lysis and centrifugation remains a crucial challenge. In this study, we developed a novel cancer cell separation chip, "hybrid double-spiral chip", that involves the serial combination of two different Dean flow fractionation (DFF) separation modes of half and full Dean cycles, which is the hybrid DFF separation mode for ultra-high-throughput blood processing at high precision and size-resolution separation. The chip allows fast processing of 5 mL whole blood within 30 min without RBC lysis and centrifugation. RBC and white blood cell (WBC) depletion rates of over 99.9% and 99%, respectively, were achieved. The average recovery rate of spiked A549 cancer cells was 87% with as low as 200 cells in 5 mL blood. The device can achieve serial reduction in the number of cells from approximately 1010 cells of whole blood to 108 cells, and subsequently to an order of 106 cells. The developed method can be combined with measurements of all recovered cells using imaging flow cytometry. As proof of concept, CTCs were successfully enriched and enumerated from the blood of metastatic breast cancer patients (N = 10, 1-69 CTCs per 5 mL) and metastatic prostate cancer patients (N = 10, 1-39 CTCs per 5 mL). We believe that the developed method will be beneficial for automated clinical analysis of rare CTCs from whole blood.

Surface acid etching for efficient anchoring of potassium on 3DOM La0.8Sr0.2MnO3 catalyst: An integration strategy for boosting soot and NOx simultaneous elimination.

The emission of soot and NOx is one of the most severe environmental issues, and the key factor is the development of catalysts in after-treatment systems. In this study, an innovative non-noble metal catalyst, named HKLSM, was fabricated by etching 3DOM La0.8Sr0.2MnO3 with citric acid and synchronously anchoring potassium salt, for soot and NOx simultaneous removal. The citric acid could not only slightly erode the 3DOM skeleton, thereby beneficial to the dispersion of potassium, but also react with high-valence state Mn to generate abundant coordination unsaturated Mn3+ sites, which could produce more active oxygen species. Moreover, HKLSM showed a higher NOx adsorption capability than the samples that were not subjected to acid etching. This adsorbed NOx could be stored as NO3- species, which could facilitate soot combustion. Among all the as-prepared catalysts, HKLSM demonstrated a competitive soot combustion activity with a T50 value of 368 °C, a TOF value of 3.24 × 10-4 s-1, a reaction rate of 1.87 × 10-7 molg-1s-1, a total NOx to N2 yield of 42.0% and favorable reusability and water-resistance. This integration strategy can rationalize an alternative protocol to soot and NOx simultaneous elimination or even other catalysis systems.

Potassium promoted macro-mesoporous Co3O4-La0.88Sr0.12CoO3-δ nanotubes with large surface area: A high-performance catalyst for soot removal.

The construction of porous perovskite nanotubular materials with a good intrinsic activity, as well as a greater dispersion of the active sites is an effective strategy to obtain a high-performance catalyst used in soot removal. Thence, macro-mesoporous Co3O4-La0.88Sr0.12CoO3-δ nanotubes with large specific surface area (154.4 m2·g-1) from the acid etching of the porous La0.6Sr0.4CoO3-δ nanotubes, are supported by 5% K through bubbling method following calcination for soot combustion. The relationship between the specific surface area and K dispersion and their effect on the activity are studied by a series of isothermal kinetic measurements combined with the characterizations and activity evaluation results. It can be found that the greater the amount is of K+ incorporated into perovskite lattice, the better the dispersion of K, as well as the La2O2CO3 formed on the catalyst surface, thus leading to the enhanced performance in the soot catalytic combustion. As a result, the 5% K supported macro-mesoporous Co3O4-La0.88Sr0.12CoO3-δ nanotubes after acid etching show good activity and stability, where the T50 is 338 °C (5% O2 + 500 ppm NO + 6% H2O) with a good CO2 selectivity (above 99%), the activate energy is 78.1 kJ·mol-1, and the turnover frequency is 5.14 × 10-4 s-1.

Surface engineering on porous perovskite-type La0.6Sr0.4CoO3-δ nanotubes for an enhanced performance in diesel soot elimination.

The porous perovskite-type La0.6Sr0.4CoO3-δ nanotubes are synthesized by sol-gel method combined with electrospinning technique following the calcination, while the porous nanotubular structure can increase the utilization of active sites related to the catalytic activity in soot oxidation. In order to further improve the catalytic activity, porous La0.6Sr0.4CoO3-δ nanotubes are further treated with nitric acid to obtain a larger specific surface area in this work. The as-prepared catalysts are characterized by different techniques to study their physical and chemical properties. The soot catalytic activity is evaluated by the temperature programmed oxidation tests and the values of activation energy. Based on the characterizations and catalytic activity evaluation, the correlation between the specific surface area and catalytic activity is well revealed by the isothermal kinetic measurements. The higher specific surface area (more than 150.0 m2 g-1) contributes to a larger amount and a better dispersion of the active oxygen species, thence improving the catalytic activity of soot oxidation. As a result, porous perovskite-type La0.6Sr0.4CoO3-δ nanotubes after nitric acid treatment for 4 h have the best activity and a good stability, with the T50 of 442 °C (5% O2) and 415 °C (5% O2 + 500 ppm NO), and the Ea of 93.6 kJ mol mol-1.

Detection and prognostic relevance of circulating tumour cells (CTCs) in Asian breast cancers using a label-free microfluidic platform.

OBJECTIVES: We aimed to study the prevalence of CTCs in breast cancer (BC) patients undergoing neoadjuvant or palliative therapy with a label-free microfluidic platform (ClearCell FX), and its prognostic relevance in metastatic BC (mBC). MATERIALS AND METHODS: Peripheral blood samples were collected from 108 BC patients before starting a new line of treatment ("baseline"), majority of whom had mBC (76/108; 70.4%). CTCs were retrieved by dean flow fractionation that enriched for larger cells, and enumerated using immunofluorescence-based staining. Progression-free survival (PFS) in mBC patients was analysed using Kaplan-Meier method; cox proportional hazard models were used for univariable and multivariable analyses. RESULTS: The detection rate of CTCs before starting a new line of treatment was 75.9% (n = 108; median: 8 CTCs/7.5 ml blood) at a cut off of ≥2 CTCs. PFS was inferior for mBC patients with baseline CTC count ≥5 CTCs/7.5 ml blood vs. those with < 5 CTCs/7.5 ml blood (median PFS: 4.3 vs. 7.0 months; p-value: 0.037). The prognostic relevance of CTCs was most significant in patients with HER2- mBC (median PFS: 4.1 vs. 8.3 months; p-value: 0.032), luminal (HR+HER2-) subtype (median PFS: 4.2 vs. 8.3 months; p-value: 0.048), and patients who had one or more prior treatments (median PFS: 4.2 vs. 7.0 months; p-value: 0.02). On multivariable analysis, baseline CTC level (hazard ratio (HR): 1.84, p-value: 0.02) and pre-treatment status (HR: 1.87, p-value: 0.05) were independent predictors of PFS. CONCLUSIONS: This work demonstrates the prognostic significance of CTCs in mBC detected using a label-free size-based enrichment platform.

ClearCell® FX, a label-free microfluidics technology for enrichment of viable circulating tumor cells.

Circulating tumor cells (CTCs) dissociate from primary tumor into the bloodstream, and carry with them cancer's fingerprints as well as the potential to turn aggressive and metastasize. In order to understand CTCs and develop clinical utility, different methods of enrichment and isolation of CTCs can be used. Here, we report the use of a label-free platform, ClearCell® FX which isolates CTCs by their mechanical features and its advantages. The technology utilizes Dean Flow Fractionation (DFF) principle in a spiral microfluidics system to separate the larger CTCs from smaller blood cells. The gentle and fast workflow allows for a range of downstream assays to be performed on the intact CTCs, particularly studies that examine an epithelial cell adhesion molecular (EpCAM)-independent population. Viable, intact cells are also retrievable for development of culture or in vivo models. © 2018 International Society for Advancement of Cytometry.

Interleukin-6 suppression reduces tumour self-seeding by circulating tumour cells in a human osteosarcoma nude mouse model.

Tumour self-seeding by circulating tumour cells (CTCs) enhances tumour progression and recurrence. Previously, we demonstrated that tumour self-seeding by CTCs occurs in osteosarcoma and revealed that interleukin-6 (IL-6) may promote CTC attraction. Here, we investigated the underlying mechanisms of IL-6 in tumour self-seeding by CTCs. IL-6 suppression inhibited in vitro cell proliferation, migration, and invasion. In addition, rhIL-6 activated the Janus-activated kinase/signal transducers and activators of transcription 3 (JAK/STAT3) and mitogen-activated protein kinase/extracellular-signal regulated kinase1/2 (MAPK/ERK1/2) pathways in vitro. Both pathways increased cell proliferation, but only the JAK/STAT3 pathway promoted migration. Suppressing IL-6 inhibited in vivo tumour growth and metastasis. IL-6 suppression or JAK/STAT3 pathway inhibition reduced CTC seeding in primary tumours. Collectively, IL-6 promotes tumour self-seeding by CTCs in a nude mouse model. This finding may provide a novel strategy for future therapeutic interventions to prevent osteosarcoma progression and recurrence.

Aberrant CXCR4 and ß-catenin expression in osteosarcoma correlates with patient survival.

To determine the clinical significance of C-X-C chemokine receptor type 4 (CXCR4) and ß-catenin in osteosarcoma, their protein expression levels were assessed in 96 osteosarcoma and 20 osteochondroma cases using immunohistochemistry. Additionally, CXCR4 and ß-catenin mRNA expression levels were measured in 16 fresh osteosarcoma and 16 adjacent healthy tissue samples using fluorescent reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In the osteosarcoma samples, the positive CXCR4 protein expression rate was significantly higher than the rate in the osteochondroma samples (68.75 vs. 20.00%; P<0.01). Furthermore, ß-catenin protein expression was detected in 61.46% of osteosarcoma cases and 25.00% of osteochondroma cases. Similarly, the RT-qPCR data identified increased CXCR4 and ß-catenin mRNA expression levels in the osteosarcoma compared with adjacent control tissues. It was determined that CXCR4 (P<0.01) and ß-catenin (P<0.05) expression were significantly associated with the clinical Enneking stage, metastasis and survival of osteosarcoma. Furthermore, multivariate analysis identified CXCR4 and ß-catenin protein expression levels, as well as clinical stage and metastasis, as significant risk factors for survival in patients with osteosarcoma (P<0.05). In conclusion, the present study determined that CXCR4 and ß-catenin are abnormally expressed in osteosarcoma tissues, and, therefore, may be important during osteosarcoma progression.

CXCR4-targeted near-infrared imaging allows detection of orthotopic and metastatic human osteosarcoma in a mouse model.

CXCR4 is expressed at primary and metastatic sites of osteosarcoma. We developed a novel CXCR4-targeted near-infrared (NIR) fluorescent imaging agent (referred to as CXCR4-IR-783). The binding to representative osteosarcoma cells (F5M2 and F4 for high- and low- CXCR4 expression) was examined. CXCR4-IR-783 fluorescence was also examined in a mouse xenograft model of human osteosarcoma using NIR fluorescence microscopy and a Kodak in-vivo multispectral system. Pulmonary metastases in mice bearing osteosarcoma xenografts were detected by micro CT, (18)F-PET scan and NIR imaging scan. Briefly, the binding of CXCR4-IR-783 was significantly higher in F5M2 than in F4 cells. Intense NIR fluorescence signals were detected in osteosarcoma xenografts, with signal/background ratio at 4.87 in mice bearing the F5M2 cell. At 4 weeks after F5M2 cell inoculation, metastatic lesions in the lungs were detectable using CXCR4-IR-783 and micro-CT scan, but not with (18)F-FDG PET scan. In conclusion, CXCR4-IR-783 is a promising tool for detection of high CXCR4-expressing osteosarcoma, and particularly for its metastatic lesions.

SDF-1/CXCR4 promotes F5M2 osteosarcoma cell migration by activating the Wnt/ß-catenin signaling pathway.

Osteosarcoma (OS), the most common primary malignant bone tumor in children and adolescents, lacks an effective therapy. Stromal cell-derived factor (SDF-1) and its receptor, CXCR4, play multiple roles in migration, proliferation, and survival of different tumor cells. This study aimed to investigate whether the functional SDF-1/CXCR4 signaling mediates chemotaxis in F5M2 OS cells as well as the underlying mechanisms. Immunohistochemistry and immunofluorescence microscopy were used. RNA expression was detected by real-time quantitative polymerase chain reaction, and protein expression was examined by Western blotting. Migration assays were carried out in F5M2 cells. The results showed that the expression of CXCR4 and ß-catenin mRNA and protein was significantly higher in OS tissues compared to the surrounding non-neoplastic tissues. SDF-1 promoted F5M2 cell migration by activating the AKT and Wnt/ß-catenin signaling pathway, which was abrogated by preincubation with AMD3100 and LY294002. In conclusion, SDF-1/CXCR4 axis-promoted F5M2 cell migration was regulated by the Wnt/ß-catenin signaling pathway.

The HIF-1α/CXCR4 pathway supports hypoxia-induced metastasis of human osteosarcoma cells.

HIF-1α mediates hypoxia-induced expression of the chemokine receptor CXCR4 and contributes to metastasis in many different cancers. We have previously shown that hypoxia promotes migration of human osteosarcoma cells by activating the HIF-1α/CXCR4 pathway. Here, immunohistochemical analysis showed that unlike control osteochondroma samples, osteosarcoma specimens were characterized by elevated expression levels of HIF-1α and CXCR4. Moreover, we found that hypoxia-induced invasiveness was more pronounced in high metastatic potential F5M2 osteosarcoma cells than in low metastatic potential F4 cells, and that this induction was sensitive to treatment with the CXCR4 antagonist AMD3100 and the HIF-1α inhibitor KC7F2. Interestingly, hypoxia-induced CXCR4 expression persisted after cultured osteosarcoma cells were returned to normoxic conditions. These observations were confirmed by experiments in a mouse model of osteosarcoma lung metastasis showing that hypoxia stimulation of pulmonary metastasis was greater in F5M2 than in F4 cells, and was sensitive to treatment with AMD3100. Our study provides further evidence of the contributions of hypoxia and the HIF-1α/CXCR4 pathway to the progression of osteosarcoma, and suggests that this axis might be efficiently leveraged in the development of novel osteosarcoma therapeutics.

In vivo molecular imaging of gastric cancer in human-murine xenograft models with confocal laser endomicroscopy using a tumor vascular homing peptide.

The early detection of premalignant lesions and cancers are very important for improving the survival of patients with gastric malignancies. Confocal laser endomicroscopy (CLE) is a novel imaging tool for achieving real-time microscopy during the ongoing endoscopy at subcellular resolution. In the present study, to evaluate the feasibility of real-time molecular imaging of GEBP11 by CLE in gastric cancer, CLE was performed on two types of tumor-bearing mice models, as well as surgical specimens of patients with gastric cancer, after the application of GEBP11. A whole-body fluorescent imaging device was first used to screen for the strongest specific fluorescent signal in xenograft models. Next, the tumor sites, as well as human tissues, were scanned with CLE. After this, targeted specimens were obtained for fluorescence microscopy and histology. We confirmed that GEBP11 could specifically bind to co-HUVECs by means of CLE in cell experiments. Thereafter, a specific signal was observed in both subcutaneous and orthotopic xenograft models in vivo after the injection of FITC-GEBP11 via tail vein, whereas the group injected with FITC-URP showed no fluorescent signals. In human tissues, a specific signal of GEBP11 was observed in 26/28 neoplastic specimens and in 8/28 samples of non-neoplastic specimens from the patients (p < 0.01). The findings from ex vivo immunofluorescence microscopy of cryostat sections correlated well with that obtained by CLE. These findings indicate that the peptide, GEBP11, might be a potential candidate for the molecular imaging of gastric cancer.

Bone marrow regeneration promoted by biophysically sorted osteoprogenitors from mesenchymal stromal cells.

Human tissue repair deficiencies can be supplemented through strategies to isolate, expand in vitro, and reimplant regenerative cells that supplant damaged cells or stimulate endogenous repair mechanisms. Bone marrow-derived mesenchymal stromal cells (MSCs), a subset of which is described as mesenchymal stem cells, are leading candidates for cell-mediated bone repair and wound healing, with hundreds of ongoing clinical trials worldwide. An outstanding key challenge for successful clinical translation of MSCs is the capacity to produce large quantities of cells in vitro with uniform and relevant therapeutic properties. By leveraging biophysical traits of MSC subpopulations and label-free microfluidic cell sorting, we hypothesized and experimentally verified that MSCs of large diameter within expanded MSC cultures were osteoprogenitors that exhibited significantly greater efficacy over other MSC subpopulations in bone marrow repair. Systemic administration of osteoprogenitor MSCs significantly improved survival rates (>80%) as compared with other MSC subpopulations (0%) for preclinical murine bone marrow injury models. Osteoprogenitor MSCs also exerted potent therapeutic effects as "cell factories" that secreted high levels of regenerative factors such as interleukin-6 (IL-6), interleukin-8 (IL-8), vascular endothelial growth factor A, bone morphogenetic protein 2, epidermal growth factor, fibroblast growth factor 1, and angiopoietin-1; this resulted in increased cell proliferation, vessel formation, and reduced apoptosis in bone marrow. This MSC subpopulation mediated rescue of damaged marrow tissue via restoration of the hematopoiesis-supporting stroma, as well as subsequent hematopoiesis. Together, the capabilities described herein for label-freeisolation of regenerative osteoprogenitor MSCs can markedly improve the efficacy of MSC-based therapies.

Slanted spiral microfluidics for the ultra-fast, label-free isolation of circulating tumor cells.

The enumeration and characterization of circulating tumor cells (CTCs), found in the peripheral blood of cancer patients, provide a potentially accessible source for cancer diagnosis and prognosis. This work reports on a novel spiral microfluidic device with a trapezoidal cross-section for ultra-fast, label-free enrichment of CTCs from clinically relevant blood volumes. The technique utilizes the inherent Dean vortex flows present in curvilinear microchannels under continuous flow, along with inertial lift forces which focus larger CTCs against the inner wall. Using a trapezoidal cross-section as opposed to a traditional rectangular cross-section, the position of the Dean vortex core can be altered to achieve separation. Smaller hematologic components are trapped in the Dean vortices skewed towards the outer channel walls and eventually removed at the outer outlet, while the larger CTCs equilibrate near the inner channel wall and are collected from the inner outlet. By using a single spiral microchannel with one inlet and two outlets, we have successfully isolated and recovered more than 80% of the tested cancer cell line cells (MCF-7, T24 and MDA-MB-231) spiked in 7.5 mL of blood within 8 min with extremely high purity (400-680 WBCs mL(-1); ~4 log depletion of WBCs). Putative CTCs were detected and isolated from 100% of the patient samples (n = 10) with advanced stage metastatic breast and lung cancer using standard biomarkers (CK, CD45 and DAPI) with the frequencies ranging from 3-125 CTCs mL(-1). We expect this simple and elegant approach can surmount the shortcomings of traditional affinity-based CTC isolation techniques as well as enable fundamental studies on CTCs to guide treatment and enhance patient care.