Activating α7nAChR helps post-myocardial infarction healing by regulating macrophage polarization via the STAT3 signaling pathway.

BACKGROUND: Monocytes/macrophages play critical roles in inflammation and cardiac remodeling following myocardial infarction (MI). The cholinergic anti-inflammatory pathway (CAP) modulates local and systemic inflammatory responses by activating α7 nicotinic acetylcholine receptors (α7nAChR) in monocytes/macrophages. We investigated the effect of α7nAChR on MI-induced monocyte/macrophage recruitment and polarization and its contribution to cardiac remodeling and dysfunction. METHODS: Adult male Sprague Dawley rats underwent coronary ligation and were intraperitoneally injected with the α7nAChR-selective agonist PNU282987 or the antagonist methyllycaconitine (MLA). RAW264.7 cells were stimulated with lipopolysaccharide (LPS) + interferon-gamma (IFN-γ) and treated with PNU282987, MLA, and S3I-201 (a STAT3 inhibitor). Cardiac function was evaluated by echocardiography. Masson's trichrome and immunofluorescence were used to detect cardiac fibrosis, myocardial capillary density, and M1/M2 macrophages. Western blotting was used to detect protein expression, and the proportion of monocytes was measured using flow cytometry. RESULTS: Activating the CAP with PNU282987 significantly improved cardiac function and reduced cardiac fibrosis and 28-day mortality after MI. On days 3 and 7 post-MI, PNU282987 reduced the percentage of peripheral CD172a + CD43low monocytes and the infiltration of M1 macrophages in the infarcted hearts, whereas it increased the recruitment of peripheral CD172a + CD43high monocytes and M2 macrophages. Conversely, MLA exerted the opposite effects. In vitro, PNU282987 inhibited M1 macrophage polarization and promoted M2 macrophage polarization in LPS + IFN-γ-stimulated RAW264.7 cells. These PNU282987-induced changes in LPS + IFN-γ-stimulated RAW264.7 cells were reversed by administering S3I-201. CONCLUSION: Activating α7nAChR inhibits the early recruitment of pro-inflammatory monocytes/macrophages during MI and improves cardiac function and remodeling. Our findings suggest a promising therapeutic target for regulating monocyte/macrophage phenotypes and promoting healing after MI.

IL-18/IL-18R1 promotes circulating fibrocyte differentiation in the aging population.

BACKGROUND: Fibrosis in multiple organs increases with age. Circulating fibrocytes are bone-marrow-derived mesenchymal progenitors that contribute to heart, lung, and kidney fibrosis under the diseased conditions. Whether circulating fibrocytes contribute to aging-related fibrosis is very limited. METHODS AND RESULTS: We measured the proportion and differentiation of circulating fibrocytes (CD45+/CD34+/collagen I+) from elders (n = 12) and adults (n = 12) using flow cytometry. Differentiated fibrocytes in the culture dishes were isolated and microarray was performed. The percentage of circulating fibrocytes in elders (1.95 ± 0.43%) was comparable to that in the adults (1.71 ± 0.38%). Cultured fibrocytes displayed enhanced potential of differentiation in the elder group (67.91 ± 5.88%) vs the adult group (44.03 ± 7.98%). In addition, expression of fibroblast activation markers and cell migratory ability were also increased in differentiated fibrocytes from elders. Microarray analysis revealed that differentiated fibrocytes from elders expressed high level of interleukin-18 (IL-18) receptor 1 (IL-18R1). Furthermore, we found IL-18 was elevated in the plasma of elders and IL-18/IL-18R1 was shown to promote fibrocyte differentiation. CONCLUSION: Circulating fibrocytes from elders had an enhanced capacity to differentiate into myofibroblasts, and might contribute to age-dependent fibrosis. Age-dependent increment of differentiation at least in part arose from their enhanced expression of IL-18R1. Inhibiting fibrocyte differentiation might be useful as an adjuvant treatment to delay the fibrosis process in aging population.

Treatment and outcomes of tumor-induced osteomalacia associated with phosphaturic mesenchymal tumors: retrospective review of 12 patients.

BACKGROUND: Tumor-induced osteomalacia (TIO) is a rare paraneoplastic syndrome characterized by severe hypophosphatemia and osteomalacia. Nonspecific symptoms make the diagnosis elusive. In addition, locating the responsible tumor(s) is challenging. The aim of this study was to investigate the clinical management and outcomes of TIO. METHODS: The clinical features, diagnostic procedures, treatment, and outcomes of 12 patients were reviewed retrospectively. RESULTS: The cohort comprised six men and six women (mean age 45.5 ± 9.9 years, range 23-61 years). The mean duration of disease was 3.7 ± 2.6 years. All patients manifested progressive bone pain, muscle weakness, and/or difficulty walking. Serum phosphorus concentrations were low in all patients (mean 0.42 ± 0.12 mmol/L). Technetium-99m octreotide scintigraphy was performed in 11 patients and showed lesions in the right distal femur, left femoral head, and right tibial plateau, respectively, in three patients. Magnetic resonance imaging (MRI) was negative for lesions in one patient. Two patients underwent biopsies that showed negative histopathology. Two patients, at 2 years and 8 months, respectively, after having negative technetium-99m octreotide studies, underwent 18F-fluorodeoxyglucose positron emission tomography/computed tomography (CT), which revealed lesions in the sacrum and soft tissue of the left palm, respectively. One tumor was detected by CT and MRI. Overall, lesion sites were the head (two patients, 16.7%), thoracic and lumbar region (two, 16.7%), pelvis (three, 25%), lower limbs (four, 33.3%), and upper limbs (one, 8.3%). All patients underwent surgery, and histopathology showed phosphaturic mesenchymal tumors in each. Postoperatively, serum phosphorus concentrations normalized within 2-7 days in 11 patients. With follow-ups of 1-41 months, surgery was effective in 10 patients. One patient developed local recurrence and another had metastases. CONCLUSIONS: Locating tumors responsible for tumor-induced osteomalacia is often challenging. Although complete tumor resection confers a good prognosis in most patients, surveillance for recurrence and metastasis is necessary. Before surgery or when surgery is not indicated, oral phosphate can alleviate symptoms and metabolic imbalance.

Predicting DNA binding proteins using support vector machine with hybrid fractal features.

DNA-binding proteins play a vitally important role in many biological processes. Prediction of DNA-binding proteins from amino acid sequence is a significant but not fairly resolved scientific problem. Chaos game representation (CGR) investigates the patterns hidden in protein sequences, and visually reveals previously unknown structure. Fractal dimensions (FD) are good tools to measure sizes of complex, highly irregular geometric objects. In order to extract the intrinsic correlation with DNA-binding property from protein sequences, CGR algorithm, fractal dimension and amino acid composition are applied to formulate the numerical features of protein samples in this paper. Seven groups of features are extracted, which can be computed directly from the primary sequence, and each group is evaluated by the 10-fold cross-validation test and Jackknife test. Comparing the results of numerical experiments, the group of amino acid composition and fractal dimension (21-dimension vector) gets the best result, the average accuracy is 81.82% and average Matthew's correlation coefficient (MCC) is 0.6017. This resulting predictor is also compared with existing method DNA-Prot and shows better performances.

[Alkaloids from cervi cornu pantotrichum and its effect on murine splenocytes proliferation].

OBJECTIVE: To study the alkaloids of Cervi Cornu Pantotrichum and its effect on murine splenocytes proliferation. METHODS: The constituents isolation and purification from Cervi Cornu Pantotrichum was carried out by reported column chromatography including Sephadex LH-20 and MCI (CHP20P) and their structures were elucidated on the basis of spectral compounds. The method of MTT was used to examine the effects of eight alkaloids and total alkaloids content (TAC) of Cervi Cornu Pantotrichum on murine splenocytes proliferation. RESULTS: Eleven compounds were isolated from Cervi Cornu Pantotrichum, and their structures were identified as follows: uracil (1), hypoxanthine (2), uridine (3) inosine (4), guanosine (5), 2'-deoxyguanosine (6), guanine (7), thymidine (8), thymine (9), cytidine (10) and adenosine (11). By the experiment of murine splenocytes proliferation activity in vitro, the results showed that the total alkaloids, uracil and adenosine had significantly promoted the proliferation of mouse spleen cells. CONCLUSION: Compounds 4 - 11 are isolated from Cervi Cornu Pantotrichum for the first time. The total alkaloids is one of the material basis of immunomodulatory effects of Cervi Cornu Pantotrichum, and uracil and adenosine are the most active.

[Diagnosis and treatment of giant cell tumor of bone in skeletal immature patients].

OBJECTIVE: To demonstrate the common characteristics of giant cell tumor of bone in immature skeletons. METHODS: From 1989 to 2009, the 8 skeletal immature patients were pathologically diagnosed with giant cell tumor (GCT) in our department, which accounted for 1.3% (8/621) of all GCT patients in an extremity. All patients were identified with an open epiphyseal plate by retrospective review of the radiograph, CT or MRI by senior consultants. Oncological and functional outcome were followed for a mean 44.1 months. There were 5 boys and 3 girls. The mean age was 13.8 years. All cases had a primary lesion. The distal femur is the most common site involved (3 cases), followed by the proximal tibia (2 cases). The proximal humerus, the distal tibia and the distal radius accounted for 1 case respectively. Oncological and functional outcome are followed for a mean 44.1 months. RESULTS: All lesions were lytic. Six lesions involved both the epiphysis and metaphysis. Two lesions located in the metaphysis area. Six lesions were treated with extended curettage and were reconstructed with allograft and (or) bone cement. Internal fixations were used in 2 cases. Two cases were treated with segmental resection. And one was reconstructed with cement spacer and the other one with segmental allograft and internal fixation. One patient (1/6) developed a bone recurrence after extended curettage. No extremity deformity and discrepancy were found during the follow up after the curettage. No metastasis was found during the follow up. CONCLUSION: Histologically GCT occurs in skeletal immature bone has the same pathological appearance but radiologically has its unique features. These lesions share same behavior as that in adults. A low local recurrence rate and good function can be achieved after a proper surgery.

[Double-column Die-punch fractures of distal radius treated with butterfly plate fixation via Henry approach].

OBJECTIVE: To explore clinical effect of open reduction and internal fixation with Henry's approach butterfly plate in treating double-column Die-punch fractures of distal radius. METHODS: From January 2018 to June 2021, 26 patients with double-column Die-column distal radius were treated with open reduction and internal fixation through Henry's surgical approach and using distal radius volar column plate(butterfly plate), including 14 males and 12 females, aged from 20 to 75 years old with an average age of (44.2±3.4) years old. Postopertaive complications were observed, Gartland-Werley score at 12 months after opertaion was used to evaluate wrist joint function. RESULTS: All 26 patients were followed up from 10 to 18 months with an average of(13.4±0.8) months. All fractures were obtained fracture union, the time ranged from 8.5 to 15.8 weeks with an average of (11.4±0.5) weeks. All incisions healed at stageⅠwithout infection, nerve injury and internal fixation failure occurred. Postoperative Gartland-Werley score at 12 months was (3.65±0.36), and 16 patients got excellent result, 8 good and 2 moderate. CONCLUSION: Open reduction and internal fixation with butterfly plate for the treatment of double-column Die-punch fractures of the distal radius through volar Henry approach could obtain satisfactory clinical outcomes.

[Inhibitory effect of 5-fluorouracil on HT-1080 human fibrosarcoma cells in vitro].

OBJECTIVE: To evaluate the inhibitory effect of 5-fluorouracil (5-Fu) on HT-1080 human fibrosarcoma cells in vitro. METHODS: HT-1080 human fibrosarcoma cells were cultured for 3 days in the proliferation period. Then adriamycin or 5-Fu at different concentrations were used to treat these cells for 24 h, 72 h and 144 h. MTT assay was used to evaluate the cytotoxic effects through measuring optical density and calculating the inhibition rate of cell growth. Morphologic changes of the cells were observed under a phase contrast microscope. Flow cytometry (FCM) was performed to detect the changes in cell cycle and DNA ploidy in the fibrosarcoma cells treated with 5-Fu. RESULTS: 10 µg/ml 5-Fu showed an inhibition rate of 45.9% (24 h), 64.7% (72 h) and 90.6% (144 h) of the HT-1080 cell growth. 100 µg/ml 5-Fu showed an inhibition rate of 53.1% (24 h), 86.4% (72 h), 93.0% (144 h) of the HT-1080 cell growth, results similar to those in the adriamycin group. Untreated fibrosarcoma cells accounted for 67.5% in G(1) phase, 21.2% in S phase and 11.3% in G(2) phase. With the increasing drug concentrations, cells in G(1) + S phase increased rapidly and no cells in G(2) phase were observed later. The cells treated with 5-Fu showed a G(1) + S cell cycle arrest. CONCLUSIONS: 5-Fu has an antitumor activity in human fibrosarcoma HT-1080 cells in vitro, in a time-dependent and dose-dependent manner. The cytotoxity of 5-Fu at high concentrations and continuous use can induce tumor cell cycle arrested at G(1) + S phase, a similar result induced with adriamycin.

Two cases of spindle cell tumors with S100 and CD34 co-expression showing novel RAF1 fusions.

BACKGROUND: Recently, a novel group of CD34 and S100 co-expression spindle cell tumors with distinctive stromal and perivascular hyalinization harboring recurrent gene fusions involving RET, RAF1, BRAF, and NTRK1/2 gene has been identified. CASE PRESENTATION: In this study, we reported two Chinese male patients with soft tissue tumors presenting in the right knee joint and the left thigh, respectively. For both patients, the tumors were completely excised with clear margin. Microscopically, case 1showed morphological overlap with neurofibroma, and case 2 showed overlap with lipomatous solitary fibrous tumor. Both tumors showed co-expression of S100 and CD34, and absence of SOX10. Genomic profiling with DNA-based next-generation sequencing (NGS) assay was performed and revealed KIF5B-RAF1 (K16:R8) and TLN2-RAF1 (T54:R8) rearrangements. RNA-based NGS and RT-PCR were performed to confirm the gene fusion. CONCLUSIONS: Though systemic therapy was not indicated in these two patients, identification of targetable kinase fusions may help to refine tumors with an ambiguous immunoprofile, and provides suggestions for targeted therapy in rare aggressive cases.

Phase I study of pegylated liposomal doxorubicin and cisplatin in patients with advanced osteosarcoma.

PURPOSE: The repeated use of doxorubicin is limited due to dose-limiting cardiac toxicity. Pegylated liposomal doxorubicin (PEG-LD, Duomeisu) has a reduced cardiac toxicity. This phase I study aimed to investigate the maximum tolerated doses (MTDs) and dose-limiting toxicities (DLTs) of the PEG-LD and cisplatin combination in patients with metastatic and recurrent osteosarcoma. METHODS: Patients were given PEG-LD at a dose of 40, 50, or 60 mg/m2 on day 1 of each 21-day cycle, according to a 3 + 3 approach for dose escalation. Cisplatin was administered as a fixed dose of 100 mg/m2 for every cycle. Toxicities and tumor response were observed. RESULTS: A total of 15 patients were enrolled in this trial, and nine of the patients had received prior doxorubicin. The MTD of PEG-LD was reached at 50 mg/m2 in this regimen, with neutropenic fever and stomatitis as DTLs. The main adverse event (AE) was myelosuppression. The most common non-hematological AEs were vomiting, hypoproteinemia, stomatitis and transient sinus arrhythmia. Grade 3-4 toxicity was neutropenia, leukopenia, thrombocytopenia, anemia and stomatitis in the whole cohort. All the AEs were relieved after symptomatic and supportive treatment. Totally, the overall response rate was 13.3% and disease control rate was 66.7%. For the six patients who have not received prior doxorubicin, one partial response and five stable diseases were observed. CONCLUSION: We provide the data showing that PEG-LD 50 mg/m2 combined with cisplatin 100 mg/m2 demonstrated an acceptable safety profile and promising clinical activity in advanced osteosarcoma, which merits further evaluation in phase II studies. TRIAL REGISTRATION: ChiCTR1900021550.

[Clinicopathologic features of primary osteosarcoma in elderly patients].

OBJECTIVE: To study the clinical manifestations, radiologic findings, pathologic diagnosis and differential diagnosis of primary osteosarcoma in elderly patients. METHODS: Twelve cases of primary osteosarcoma occurring in patients older than 60 years were encountered during the period from 1985 to 2010. The clinical manifestations, radiologic features and pathologic findings were studied and the follow-up data were analyzed. RESULTS: The sites of involvement included long bones (number = 7), ilium (number = 1), craniofacial bones (number = 2) and soft tissue (number = 2). Radiologic examination showed a mixture of osteosclerotic and osteolytic lesions in 10 patients, soft tissue lesions with high-density areas in 2 patients and soft tissue lesions with periosteal reaction in 8 patients. Histologically, most cases showed features of conventional osteosarcoma. There were 2 cases of malignant fibrous histiocytoma-like osteosarcoma, 2 cases of chondroblastic osteosarcoma and 1 case of well-differentiated intraosseous osteosarcoma. Immunohistochemical study played little role in pathologic diagnosis. Ten patients had undergone amputation, including one patient who had received adjuvant chemotherapy beforehand. Nine patients had follow-up information available. Three of them died of lung metastasis and 1 died of cardiovascular disease. CONCLUSIONS: Primary osteosarcoma rarely occurs in elderly patients and can easily be missed. Correlation with clinical, radiologic and histologic features is important for arriving at a correct diagnosis.

[Retrospective analysis and prognostic factors of 208 cases of primary soft tissue sarcoma of extremity].

OBJECTIVE: To investigate the prognosis factors of soft tissue sarcoma, especially the impact of surgical treatment on the prognosis. METHODS: We retrospectively reviewed 208 surgically treated patients. There were 128 male and 80 female. The average age was 46 ranged from 9 to 98 years old. Possible factors of whether the patient firstly treated in our hospital, the tumor size (< 5 cm, 5 ∼ 10 cm, > 10 cm), tumor depth (superficial deep fascia, under the deep fascia), histological type (such as adipose sarcoma, malignant fibrous histiocytoma, synovial sarcoma, fibrous sarcoma, malignant peripheral nerve sheath tumors, other tumors), tumor grade (FNCLCC I, II, III), surgical margin (intralesional, marginal, wide, radical) and adjuvant therapy on the prognosis of patients were analyzed. RESULTS: The median follow-up was 37.5 ranged from 1.3 to 128.1 months. The overall 3-year and 5-year survival were 77% and 75%. The overall 3-year and 5-year recurrence rate were 28% and 37%. The overall 3-year and 5-year metastasis rate were 35% and 43%. Tumor size, tumor grade and metastasis or not independently affected survival (χ(2) = 18.813, 24.849 and 21.107, all P < 0.05). Whether the patient firstly treated in our hospital and histological type independently affect the local recurrence (χ(2) = 21.915, 12.192, both P < 0.05); histological grade can independently affect the metastasis (χ(2) = 7.714, P < 0.05). Surgical margin alone affected the local recurrence and metastasis (χ(2) = 19.610, 9.272, both P < 0.05). CONCLUSIONS: Surgical margin independently affected local recurrence and distant metastasis, and thus indirectly affect the survival of soft tissue sarcoma. In particular, the primary choice for treatment of soft tissue sarcoma without metastasis should be surgery. Wide or radical margin could significantly improve the prognosis of soft tissue sarcoma patients.

[Surgical treatment of massive soft tissue sarcoma in the shoulder girdle].

OBJECTIVE: To detect the character of surgical treatment of massive soft tissue sarcoma in the shoulder girdle and analyze the impact factor to the result. METHODS: Seven patients with massive soft tissue sarcoma in the shoulder girdle were treated in our department between 2005 and 2009. There were 4 males and 3 females. All the patients were referred to our hospital after local recurrence post-operatively. The mean age was 43.8 years old (range 14 - 75). The maximum diameter of the tumor varied from 10 to 16 centimeters. All the patients were performed surgery, wide margin in 4 cases and marginal margin in 3 cases. Five were performed tumor resection and reconstruction with latissimus dorsi muscle flap transfer and skin graft. One was reconstructed with advanced skin flap and skin graft. The other one was treated with skin graft. The diagnosis included 3 malignant fibrous histiocytomas, 1 low grade myxoid fibrosarcoma, 1 Primitive neuroectodermal tumor, 1 rhabdomyosarcoma, 1 dermatofibrosarcomas protuberans. The MSTS score system was used to evaluate the shoulder function. RESULTS: Seven patients were followed up with long time. The mean follow up was 29 months (range 10 to 46 months). Two patients suffered local recurrence and one died of pulmonary metastasis 6 months after the second surgery for local recurrence. One patient suffered pulmonary metastasis. The last four patients were disease-free at the end of follow-up. The function of shoulder girdle was satisfactory. The mean MSTS score was 28. CONCLUSIONS: Soft tissue sarcomas in the shoulder girdle are easy to be misdiagnosed and mistreated. Wide surgical margin was the key impact factor to the local recurrence of soft tissue sarcoma in the shoulder girdle. The surgical margin and invasion of the tumor are the key factor to the prognosis. The soft tissue defect after surgery is often reconstructed by muscle flap transfer or skin flap transfer. The latissimus dorsi muscle flap transfer is often used.

[Computer navigation-guided excision of osteoid osteomas].

OBJECTIVE: To report the experience for the precision osteoid osteoma resection using computer navigation system. METHODS: Between January 2008 and December 2009, 26 surgical resections were performed for 26 patients who had osteoid osteoma with computer navigation system. There were 23 males and 3 females with an average age of 18 years (7 to 35). Tumors were located at femoral shaft 9, femoral trochanter 4, femoral neck 2, tibial shaft 5, metaphysic of proximal tibia 1, acetabulum 2, pubis 1, vertebral appendix 1 and radial shaft 1. Pre-operative X-ray and CT of each patient was performed to confirm the diagnosis. It was carried out intraoperatively the process of CT-based navigation in 4 cases and intraoperative Iso-C three-dimensional navigation in 22 cases. The Navigation System software was Spine Navigation 1.2 in all cases. The Pointer was helpful to localize the lesion and precisely resected the lesion without removal of any excess bone. RESULTS: All the navigation operations were finished successfully with curettage for 12 and En Bloc resection for 14. Bone grafting was made in 21 cases and none in 3 cases. The completely clearance of nidus by intraoperative visual inspection and Pointer confirmation, postoperative X-ray and(or) CT scan was performed in all cases. All cases had histopathology diagnosis of osteoid osteoma and immediate pain relief after surgery. All cases were followed up for 20.6 months averaged (12 to 35 months). No local recurrence and pain relapse occurred. CONCLUSIONS: The navigation system is very helpful for the precision tumor resection of nidus. Especially for the patients with osteoid osteoma located at diaphysis, Intraoperative Iso-C three-dimensional navigation is more useful.

Computer navigation-aided joint-preserving resection and custom-made endoprosthesis reconstruction for bone sarcomas: long-term outcomes.

BACKGROUND: Computed tomography (CT) and magnetic resonance imaging (MRI) data can be fused to identify the tumor boundaries. This enables surgeons to set close but tumor-free surgical margins and excise the tumor more precisely. This study aimed to report our experience in performing computer navigation-aided joint-preserving resection and custom-made endoprosthesis reconstruction to treat bone sarcoma in the diaphysis and metaphysis of the femur and tibia. METHODS: Between September 2008 and December 2015, 24 patients with bone sarcomas underwent surgical resection and joint-sparing reconstruction under image-guided computer navigation. The cohort comprised 16 males and eight females with a median age of 19.5 years (range: 12-48 years). The tumor location was the femoral diaphysis in three patients, distal femur in 19, and proximal tibia in two. The tumors were osteosarcoma (n = 15), chondrosarcoma (n = 3), Ewing sarcoma (n = 3), and other sarcomas (n = 3). We created a pre-operative plan for each patient using navigation system software and performed navigation-aided resection before reconstructing the defect with a custom-made prosthesis with extracortical plate fixation. RESULTS: Pathological examination verified that all resected specimens had appropriate surgical margins. The median distance from the tumor resection margin to the joint was 30 mm (range: 13-80 mm). The median follow-up duration was 62.5 months (range: 24-134 months). Of the 24 patients, 21 remain disease free, one is alive with disease, and two died of the disease. One patient developed local recurrence. Complications requiring additional surgical procedures occurred in six patients, including one with wound hematoma, one with delayed wound healing, one with superficial infection, one with deep infection, and two with mechanical failure of the prosthesis. The mean Musculoskeletal Tumor Society score at the final follow-up was 91% (range: 80%-100%). The 5- and 10-year implant survival rates were 91.3% and 79.9%, respectively. CONCLUSIONS: Computer navigation-aided joint-preserving resection and custom-made endoprosthesis reconstruction with extracortical plate fixation is a reliable surgical treatment option for bone sarcoma in the diaphysis and metaphysis of the femur and tibia.

[Result analysis of percutaneous core needle biopsy for bone tumors in upper limbs with pathological fracture].

OBJECTIVE: To analyze the results of percutaneous core needle biopsy for bone tumors in upper limbs with pathologic fracture and to find the possible factors that could impact the results. METHODS: The including criteria for this study was the patients who had received percutaneous core needle biopsy and definitive surgery, whose tumor was located at upper limb with pathologic fracture. From January 2015 to December 2019, seventy-seven patients were enrolled. There were 55 males and 22 females. The median age was 27 years old (range:5 to 88 years old). The tumor located at humerus in 67 cases, radius in 8 cases and ulna in 2 cases. If the pathologic diagnosis of core needle biopsy was the same with the definitive surgery, it was defined as "correct". If the pathologic diagnosis of biopsy for benign or malignant was right but the exact diagnostic name was not the same with definitive surgery, it was defined as "supportive". If the pathologic diagnosis of biopsy for benign or malignant was not correct, it was defined as "wrong". We retrospectively analyzed the accuracy and impact factors for core needle biopsy. RESULTS: The result was "correct" in 63 cases(81.8%), "supportive" in 14 cases(18.2%), and "wrong" in 0 cases. We analyzed the gender, age, location, fracture displacement, the destroyed type for bone tumor, soft tissue mass, fluid area in the tumor as the factors. The results showed the rate for "correct" was significantly higher when the tumor had soft tissue mass (P< 0.05) and lower when the fluid area existed inside the tumor (P<0.05). CONCLUSION: The accuracy of percutaneous core needle biopsy for upper limb bone tumor with pathologic is high and acceptable. The biopsy chosen the soft tissue mass area can increase the accuracy.

Frequent amplification of HDAC genes and efficacy of HDAC inhibitor chidamide and PD-1 blockade combination in soft tissue sarcoma.

BACKGROUND: The advent of immune checkpoint therapy has been a tremendous advance in cancer treatment. However, the responses are still insufficient in patients with soft tissue sarcoma (STS). We aimed to identify rational combinations to increase the response to immune checkpoint therapy and improve survival. METHODS: Whole-exome sequencing (WES) was performed in 11 patients with liposarcoma. Somatic copy number alterations (SCNAs) were analyzed at the gene level to identify obvious amplification patterns in drug-target genes. The expression and prognostic value of class I histone deacetylases (HDACs) was evaluated in 49 patients with sarcoma in our center and confirmed in 263 sarcoma samples from The Tumor Cancer Genome Atlas (TCGA) database. Q-PCR, flow cytometry and RNA-seq were performed to determine the correlations between class I HDACs, chidamide and PD-L1 in vitro and in vivo. The efficacy of combining chidamide with PD-1 blockade was explored in an immunocompetent murine model and a small cohort of patients with advanced sarcoma. Western blot, ChIP assay and dual luciferase assessment were applied in the mechanistic study. RESULTS: The HDAC gene family was frequently amplified in STS. SCNAs in the HDAC gene family were extensively amplified in 8 of 11 (73%) patients with liposarcoma, based on a drug-target gene set, and we verified amplification in 76.65% (197/257) of cases by analyzing TCGA sarcoma cohort. Class I HDAC expression is associated with a poor prognosis for patients with STS, and its inhibition is responsible for promoting apoptosis and upregulating of programmed cell death ligand 1 (PD-L1). The HDAC class I inhibitor chidamide significantly increases PD-L1 expression, increased the infiltration of CD8+ T cells and reduced the number of MDSCs in the tumor microenvironment. The combination of chidamide with an anti-PD-1 antibody significantly promotes tumor regression and improves survival in a murine model. Moreover, chidamide combined with the anti-PD-1 antibody toripalimab is effective in patients with advanced and metastatic sarcoma, and the side effects are tolerable. Mechanistically, chidamide increases histone acetylation at the PD-L1 gene through the activation of the transcriptional factor STAT1. CONCLUSIONS: The combination of chidamide and anti-programmed cell death 1 (PD-1) therapy represents a potentially important strategy for STS.

Feasibility of MRI Radiomics for Predicting KRAS Mutation in Rectal Cancer.

The mutation status of KRAS is a significant biomarker in the prognosis of rectal cancer. This study investigated the feasibility of MRI-based radiomics in predicting the mutation status of KRAS with a composite index which could be an important criterion for KRAS mutation in clinical practice. In this retrospective study, a total of 127 patients with rectal cancer were enrolled. The 3D Slicer was used to extract the radiomics features from the MRI images, and sparse support vector machine (SVM) with linear kernel was applied for feature reduction. The radiomics classifier for predicting the KRAS status was then constructed by Linear Discriminant Analysis (LDA) and its performance was evaluated. The composite index was determined with LDA model. Out of 127 rectal cancer subjects, there were 44 KRAS mutation cases and 83 wild cases. A total of 104 radiomics features were extracted, 54 features were filtered by linear SVM with L1-norm regularization and 6 features that had no significant correlations within them were finally selected. The radiomics classifier constructed using the 6 features featured an AUC value of 0.669 (specificity: 0.506; sensitivity: 0.773) with LDA. Furthermore, the composite index (Radscore) had statistically significant difference between the KRAS mutation and wild groups. It is suggested that the MRI-based radiomics has the potential in predicting the KRAS status in patients with rectal cancer, which may enhance the diagnostic value of MRI in rectal cancer.

Platelets Promote Ang II (Angiotensin II)-Induced Atrial Fibrillation by Releasing TGF-ß1 (Transforming Growth Factor-ß1) and Interacting With Fibroblasts.

Hypertension is a risk factor of atrial fibrillation (AF), and a certain number of patients with hypertension were found with an enlarged left atrium. Platelet activation is found in patients with hypertension or pressure overload/Ang II (angiotensin II)-induced hypertensive animal models and contribute to ventricular fibrosis. Whether hypertension-induced atrial fibrosis is mediated by platelets remains unknown. Our previous experimental data showed that platelet-derived TGF-ß1 (transforming growth factor-ß1) was reduced in patients with hypertensive AF. The present study is to investigate whether platelet-derived TGF-ß1 promotes Ang II-induced atrial fibrosis and AF. Platelet activation and atrial platelet accumulation were measured in sinus rhythm controls, normotensive AF, and patients with hypertensive AF. Ang II (1500 ng/kg per minute, 3 weeks) infused mice with pharmacological (clopidogrel) and genetic platelet inhibition (TGF-ß1 deletion in platelets) were used. Platelet activation, atrial structural remodeling, atrial electrical transmission, AF inducibility, inflammation, and fibrosis were measured in mice. We found that circulating platelets were activated in patients with hypertensive AF. A large amount of platelet was accumulated in the atriums of patients with hypertensive AF. Both clopidogrel treatment and platelet-specific deletion of TGF-ß1 attenuated Ang II-induced structural remodeling, atrial electrical transmission, AF inducibility, as well as atrial inflammation and fibrosis than mice without interventions. Furthermore, clopidogrel blocked atrial platelet accumulation and platelet-fibroblast conjugation. Platelets promoted atrial fibroblast differentiation in cell culture. Profibrotic actions of platelets are largely via activation of atrial fibroblasts by releasing TGF-ß1 and inducing platelet-fibroblast conjugation, and platelet inhibition is sufficient to inhibit atrial fibrosis and AF inducibility.

[Surgical treatment for bone nonunion after massive allograft transplantation].

OBJECTIVE: To evaluate the surgical treatment and outcome of autogenous bone grafting and internal fixation in management of bone nonunion after massive allograft transplantation. METHODS: From January 1994 to December 2006, 41 of 176 patients underwent bone nonunion after massive allograft transplantation. Twenty-two of 41 patients received autogenous bone grafting. Complete clinical and follow-up data was available for 15 cases. The average age at secondary autogenous bone grafting was 24 years old (ranging from 15 to 34). The primary diseases included osteosarcoma (5 cases), giant cell tumor (4 cases), parosteal osteosarcoma (2 cases), hemangioendothelioma (2 cases) and primitive neuroectodermal tumor (2 cases). Tumor was located at distal femur in 7 patients, middle of humerus in 3, middle of femur in 2, proximal tibia in 2 and proximal humerus in 1. Eight of 15 patients with simple bone nonunion received autogenous bone grafting. Another 7 patients with bone nonunion and fracture of primary internal fixation underwent autogenous bone grafting and re-internal fixation. RESULTS: At a mean follow-up of 46.8 months (ranging from 18 to 148 months), bone union was observed in 13 of 15 patients (86.7%) with the mean healing time 13.3 months (ranging from 5 to 20). Bone union could be observed in all 8 patients with simple bone nonunion and 5 of 7 patients with bone nonunion and internal fixation fracture, similar healing time 14 and 12 months respectively. There was no infection or any other complications. Two patients underwent re-nonunion received prosthesis replacement at last. The mean MSTS score of 13 patients was 25.1, with 8 simple bone nonunion patients and 5 combined with internal fixation fracture patients 25.4 and 24.6 respectively, also basically no difference. CONCLUSIONS: Autogenous bone grafting and internal fixation in management of nonunion after massive allograft transplantation have the advantage of easy operation, less complications, high rate of bone healing and good function result with obvious superiority to prosthesis replacement. For management of nonunion after massive allograft transplantation, autogenous bone grafting and internal fixation is mostly recommended.