Mutation characteristics of osteosarcoma: a single center study of 64 cases using next-generation sequencing / 中华病理学杂志

Objective: To investigate the distribution and characteristics of gene mutations in osteosarcoma, and to analyze the frequency and types of detectable mutations, and to identify potential targets for individualized treatment of osteosarcoma. Methods: The fresh tissue or paraffin-embedded tissue samples of 64 cases of osteosarcoma that were surgically resected or biopsied and then subject to next generation sequencing, were collected from Beijing Jishuitan Hospital, China from November 2018 to December 2021. The tumor DNA was extracted to detect the somatic and germline mutations using targeted sequencing technology. Results: Among the 64 patients, 41 were males and 23 were females. The patient age ranged from 6 to 65 years with a median age of 17 years, including 36 children (under 18 years old) and 28 adults. There were 52 cases of conventional osteosarcoma, 3 cases of telangiectatic osteosarcoma, 7 cases of secondary osteosarcoma, and 2 cases of parosteosarcoma. The detection rate of gene mutations was overall 84.4% (54/64). There were 324 variations in 180 mutated genes, including 125 genes with copy number variations, 109 single nucleotide variants, 83 insertions or deletions, and 7 gene fusions. The most common mutated genes were TP53, VEGFA, CCND3, ATRX, MYC, RB1, PTEN, GLI1, CDK4 and PTPRD. Among them, TP53 had the highest mutation rate (21/64, 32.8%), single nucleotide variant was the main mutation type (14/23, 60.9%), and 2 cases carried the TP53 germline mutation. VEGFA and CCND3 showed copy number amplification simultaneously in 7 cases. Conclusions: The high-frequency mutation of TP53 suggests that it plays an important role in the pathogenesis and development of osteosarcoma. VEGFA, CCND3 and ATRX are mutated genes in osteosarcoma and worthy of further studies. Combination of pathologic diagnosis and next generation sequencing with clinical practice can guide individualized treatment for patients with refractory, recurrent and metastatic osteosarcoma.

Magnetic Resonance Imaging for the Assessment of Long Bone Tumors / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Wide resection margins of osseous tumors are associated with a low incidence of local recurrence, making accurate measurement of the intraosseous extent of primary malignant long bone tumors is crucial. We compared the intraosseous tumor extent assessed by magnetic resonance imaging (MRI) with the gross specimen to evaluate the accuracy of MRI.</p><p><b>METHODS</b>A total of 255 patients with primary malignant tumors in the long bones were included. Using MRI, we defined the length of tumor as the distance from the articular surface to the boundary between abnormal and normal marrow signal. The extent of the abnormal intraosseous signal was measured on unenhanced T1-weighted (T1WI) magnetic resonance images after chemotherapy. All gross surgical specimens were sectioned, and tumor extent was measured. Wilcoxon signed-rank test was used to test the differences between MRI and gross specimen findings. Spearman's correlation analysis was used to test the correlation between groups.</p><p><b>RESULTS</b>Median tumor length by gross specimen (112 mm; range, 45-300 mm) was longer than that by MRI (108 mm; range, 45-304 mm; Z = -6.916, P < 0.001). Of 255 images, tumor length was accurately represented on 27 T1WI magnetic resonance images, overestimated on 79 images, and underestimated on 149 images. The median difference between imaging and gross specimen measurements was 2.0 mm (range: 1.0-15.0 mm) for the 79 cases where tumor length was overestimated, and 5.0 mm (range: 1.0-18.0 mm) for the 149 cases where tumor length was underestimated. The Spearman correlation demonstrated a high correlation of tumor length on gross specimen with the tumor length on MRI (R = 0.99, P < 0.01).</p><p><b>CONCLUSIONS</b>We conclude that preoperative MRI could be a useful method in determining intramedullary malignant bone tumor boundaries and may serve as an accepted assessment method of long bone tumors before limb-sparing surgery.</p>

Computer Navigation-aided Resection of Sacral Chordomas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Resection of sacral chordomas is challenging. The anatomy is complex, and there are often no bony landmarks to guide the resection. Achieving adequate surgical margins is, therefore, difficult, and the recurrence rate is high. Use of computer navigation may allow optimal preoperative planning and improve precision in tumor resection. The purpose of this study was to evaluate the safety and feasibility of computer navigation-aided resection of sacral chordomas.</p><p><b>METHODS</b>Between 2007 and 2013, a total of 26 patients with sacral chordoma underwent computer navigation-aided surgery were included and followed for a minimum of 18 months. There were 21 primary cases and 5 recurrent cases, with a mean age of 55.8 years old (range: 35-84 years old). Tumors were located above the level of the S3 neural foramen in 23 patients and below the level of the S3 neural foramen in 3 patients. Three-dimensional images were reconstructed with a computed tomography-based navigation system combined with the magnetic resonance images using the navigation software. Tumors were resected via a posterior approach assisted by the computer navigation. Mean follow-up was 38.6 months (range: 18-84 months).</p><p><b>RESULTS</b>Mean operative time was 307 min. Mean intraoperative blood loss was 3065 ml. For computer navigation, the mean registration deviation during surgery was 1.7 mm. There were 18 wide resections, 4 marginal resections, and 4 intralesional resections. All patients were alive at the final follow-up, with 2 (7.7%) exhibiting tumor recurrence. The other 24 patients were tumor-free. The mean Musculoskeletal Tumor Society Score was 27.3 (range: 19-30).</p><p><b>CONCLUSIONS</b>Computer-assisted navigation can be safely applied to the resection of the sacral chordomas, allowing execution of preoperative plans, and achieving good oncological outcomes. Nevertheless, this needs to be accomplished by surgeons with adequate experience and skill.</p>

Is There a Reliable Method to Predict the Limb Length Discrepancy after Chemotherapy and Limb Salvage Surgery in Children with Osteosarcoma? / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>For a child with osteosarcoma, prediction of the limb length discrepancy at maturity is important when planning for limb salvage surgery. The purpose of this study was to provide a reliable prediction method.</p><p><b>METHODS</b>A retrospective review of Chinese children receiving chemotherapy for osteosarcoma before skeletal maturity was conducted. Standing full-length radiographs of the lower extremity were used for length measurements. Length-for-age curves were constructed using the LMS method. The lower limb multiplier for a specific age and gender was calculated using the formula M = Lm/L, where M was the gender- and age-specific multiplier, Lmwas the bone length at maturity, and L was the age-specific bone length. Prematurity and postmaturity radiographs were used to assess the accuracy of the prediction methods.</p><p><b>RESULTS</b>A total of 513 radiographs of 131 boys and 314 radiographs of 86 girls were used to calculate the coefficients of the multiplier. The multipliers of 8-, 9-, 10-, 11-, 12-, 13-, 14-, 15-, 16-, 17-, and 18-year-old boys after chemotherapy for osteosarcoma were 1.394, 1.306, 1.231, 1.170, 1.119, 1.071, 1.032, 1.010, 1.004, 1.001, and 1.000, respectively; while for girls at the same ages, the multipliers were 1.311, 1.221, 1.146, 1.092, 1.049, 1.021, 1.006, 1.001, 1.000, 1.000, and 1.000, respectively. Prematurity and postmaturity femoral and tibial lengths of 21 patients were used to assess the prediction accuracy. The mean prediction error was 0 cm, 0.8 cm, and 1.6 cm for the multiplier method using our coefficients, Paley's coefficients, and Anderson's method, respectively.</p><p><b>CONCLUSIONS</b>Our coefficients for the multiplier method are reliable in predicting lower limb length growth of Chinese children with osteosarcoma.</p>

Subchondral bone grafting reduces degenerative change of knee joint in patients of giant cell tumor of bone / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Giant cell tumors (GCTs) most commonly occur around the knee. The most beneficial procedure for this disease has been extensive curettage with reconstruction. However, since many GCTs may compromise the subchondral bone, surgery can further jeopardize the articular cartilage and result in secondary osteoarthritis. In this study, we aimed to determine the factors associated with the development of degenerative arthritis and the effect of bone grafting on the prevention of secondary osteoarthritis.</p><p><b>METHODS</b>We retrospectively analyzed 76 patients with GCT around the knee. The mean age at first diagnosis was 31.1 years. Surgical treatments included extensive curettage and cementation with or without bone grafting in the subchondral bone. Patient follow-up was a median duration of 35 months, ranging from 18 to 113 months.</p><p><b>RESULTS</b>The local recurrence rate was 5.3% (4/76). Secondary degenerative changes occurred in 30.3% (23/76) of the patients. Less than 10 mm of the residual thickness of the remaining subchondral bone was correlated with secondary degenerative changes in 57 patients (P < 0.001). Of these 57 patients, 56.5% (13/23) treated with bone cement reconstruction alone developed secondary degenerative changes; following bone grafting, the rate decreased to 29.4% (10/34), with a statistically significant difference (P = 0.041).</p><p><b>CONCLUSIONS</b>GCT patients with less residual thickness of the subchondral bone are more likely to develop degenerative arthritis after curettage. Bone grafting in the subchondral bone area is recommended when the residual thickness of the subchondral bone is less than 10 mm.</p>

Inhibitory effect of 5-fluorouracil on HT-1080 human fibrosarcoma cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the inhibitory effect of 5-fluorouracil (5-Fu) on HT-1080 human fibrosarcoma cells in vitro.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Retrospective analysis and prognostic factors of 208 cases of primary soft tissue sarcoma of extremity / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the prognosis factors of soft tissue sarcoma, especially the impact of surgical treatment on the prognosis.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>

Surgical treatment for bone nonunion after massive allograft transplantation / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the surgical treatment and outcome of autogenous bone grafting and internal fixation in management of bone nonunion after massive allograft transplantation.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

The status and prospect of total body irradiation technique / 中国医疗器械杂志

This paper describes the development of TBI technique about fractionated TBI and the relationship between irradiation dose rate and complication; IMRT for TBI and lung compensation technique.

Design of a 3D afterloading brachytherapy simulation system based on CT images / 中国医疗器械杂志

<p><b>OBJECTIVE</b>To design a new afterloading brachytherapy simulation system based on CT images.</p><p><b>METHODS</b>This paper mainly focuses on the anthropomorphic pelvic phantom spiled by three pipelines and the nasopharyngeal carcinoma spiled by two pipelines. Microsoft Visual C++ was used to parse CT images for some information, then to reconstruct pipelines in the body of phantom or the patient and to give the three-dimensional coordinate of dwelling points. The dose distribution displayed on CT images was processed by the dose distribution calculation methods near single afterloading source and the dose optimization methods. VTK technology was used in the 3D display in the system.</p><p><b>RESULTS</b>According to the reference points applied by doctors, the system can calculate reversely the dwelling time of dwelling points in pipelines and get satisfying dose distribution on CT images. Besides, it can reflect the 3D relationship between the dose volume and the normal tissues.</p><p><b>CONCLUSIONS</b>This system overcomes some deficiencies of 2D afterloading brachytherapy simulation system based on X-ray films which are used widely in China. It supplies 3D display of dose distribution for clinical doctors. At present, the system is being tested in clinics.</p>

Study on integrated Chinese and Western therapy and criterion for efficacy evaluation of gastroesophageal reflux disease--a clinical observation on 116 cases / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the criterion for efficacy evaluation of gastroesophageal reflux diseases (GERD) and the effect of treatment with integrated Chinese and Western medicine.</p><p><b>METHODS</b>One hundred and sixteen patients of GERD were randomly assigned to two groups, group A treated with Western medicine and group B with integrated Chinese and Western medicine. Changes of symptoms, TCM syndromes, and endoscopic picture were observed before and after treatment. And the condition of recurrence was also inspected.</p><p><b>RESULTS</b>The total effective rate was 84.5% and 67.2% in group A and B respectively. The long-term efficacy was better in group B than that in group A (P < 0.05), while there was no significant difference in short-term efficacy between the two groups. But the recurrence happened in both groups after withdrawal of treatment. The recurrence rate was significantly higher in patients of deficiency syndrome type and coexistence of deficiency and excess syndrome type than that in those of excess syndrome type (P < 0.05), and it was also higher in those whose pathogenesis was associated with mental factor (P < 0.05). In the other 20 patients the treatment was sustained for 10.4 +/- 11.4 months on average. There were 69.0% of the patients with normal esophagus mucosa shown by endoscope examination, and the pathological changes were not coincident with the symptoms and prognosis.</p><p><b>CONCLUSIONS</b>Basically, no case of GERD could be cured, so, it is supposed that the item of "cure" in the criteria of therapeutic efficacy evaluation is advisably made over to "clinical control" or "obvious efficacy", and add an item of "no change" in the criterion. The importance of endoscopic picture for efficacy evaluation is doubtful and needs to be further discussed.</p>

Three strategies for hemostasis at femoral artery access site: A comparison of safety and efficacy / 第二军医大学学报

Objective: To assess the safety and efficacy of the standard manual compression and 2 arterial puncture closing devices, Angioseal and Perclose, for hemostasis at the femoral artery access site in patients undergoing coronary angiography (CAG) or percutaneous coronary interventions (PCI). Methods: Totally 366 patients undergoing coronary angiography or PCI were assigned to receive either Angioseal (n=128), Perclose (n=110) or standard manual compression (n=128). The efficacy endpoint (immediate hemostasis, successful hemostasis rate, operating time, time of leg immobilization and time to hemostasis) and safety endpoint (vasovagal reflex, major complications, local complications, hematocrit drop, etc.) were evaluated. Risk factors (gender, age, body mass index, comorbid conditions, antiplatelet agents, and anticoagulant agents, etc.) were also analyzed. Results: The successful hemostasis rates were similar be tween Angioseal group and Perclose group, but the rate of immediate hemostasis of Angioseal group was higher than that of Perclose group (P<0.01). Time of leg immobilization and time to hemostasis in Angioseal group and Perclose group was shorter than those of manual compression group(P<0.01). Both vasovagal reflex (9.3%) and major complications (3.1%) were only noticed in manual compression group. The rates of local complications in manual compression group and Angioseal group were both lower than that of Perclose group(P<0.01). Post-procedure unfractionated heparin was associated with increased risk of hematoma (OR=4.382, P<0.05), post-procedure clopidogrel (OR=3.549, P<0.01) and interventional procedures (OR=6.584, P<0.05) were associated with increased risk of blood oozing, and female gender was associated with a reduced risk of blood oozing (OR=0.132, P<0.05). The successful hemostasis rates were lower in the elderly and those receiving PCI. Post-procedure unfractionated heparin and low molecular weight heparin was associated with longer hemostasis time. Conclusion: Perclose and An gioseal can be an alternative of standard manual compression for hemostasis of the femoral artery access site. Angioseal may be better than Perclose regarding the immediate hemostasis and local complications.

Effect of Rhein on the development of hepatic fibrosis in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of rhein on the development of hepatic fibrosis.</p><p><b>METHODS</b>The animal models were made with carbon tetrachloride (CCl(4)) mixed with vegetable oil (3/2, v/v), which was injected subcutaneously twice a week for 6 weeks, and with 5% ethanol for free drinking water. At the same time, Rhein was administrated at the dose of 25 mg/kg or 100 mg/kg once a day for 6 weeks. The changes of both biochemical markers, such as the levels of alanine aminotransferase (ALT), hyaluronic acid (HA), procollagen type III (PCIII) in serum and SOD, malondialdehyde (MDA) in liver, and related histopathological parametres were determined.</p><p><b>RESULTS</b>Compared with the model group, there were three kinds of changes in the larger quantity of rhein treated group. (1) The levels of ALT, HA, PCIII in serum and MDA in liver homogenate were decreased significantly (from 150 U/L +/- 16 U/L to 78 U/L +/- 18 U/L, 321 microg/L +/- 97 microg/L to 217 microg/L +/- 75 microg/L, 31 microg/L +/- 14 microg/L to 16 microg/L +/- 6 microg/L and 3.67 nmol/mg +/- 0.68 nmol/mg to 1.88 nmol/mg +/- 0.34 nmol/mg, respectively, t > or 2.977, P<0.01). However the level of SOD in liver was increased (from 62.45 NU/mg +/- 8.74 NU/mg to 91.26 NU/mg +/- 14.04 NU/mg, t=4.453, P<0.01). (2) The expressions of transforming growth factor beta 1 (TGF-beta 1) and alpha-smooth muscle actin (alpha-SMA) in liver were markedly reduced (P<0.05 and P<0.01). (3) The collagen staining positive area was decreased and the grade of fibrosis was reduced significantly in liver (P<0.05 and P<0.01).</p><p><b>CONCLUSION</b>Rhein can protect hepatocyte from injury and prevent the progress of hepatic fibrosis in rats, which may associate with that rhein plays a role in antioxidation, anti-inflammation, inhibiting the expression of TGF-beta1 and suppressing the activation of hepatic stellate cells (HSCs).</p>