Ifosfamide

Eleven patients hospitalized from October 2017 to April 2019 in Shanghai Sixth People’s Hospital were prospectively enrolled in the study, which was approved by the Shanghai Sixth People’s Hospital Ethics Committee. Each patient provided written informed consent. All 11 patients (five male and six female, age range from 11 to 38-years old) had been diagnosed with osteoblastic or chondroblastic OS according to the NCCN Clinical Practice Guidelines in Oncology (https://www.nccn.org/). Samples for scRNA-seq were derived from the primary tumor sites of seven patients who had received traditional first-line adjuvant and neo-adjuvant chemotherapy composed of a cocktail of four drugs (doxorubicin, cisplatin, methotrexate and ifosfamide), as well as surgical therapy. Two patients with lung metastasis and two with recurrent disease had received gemcitabine in combination with docetaxel (GT). In addition to other patients enrolled for our study, we recruited one patient (BC17) from the clinical trial NCT03676985, in which all patients had undergone neoadjuvant chemotherapy, surgery, and adjuvant chemotherapy, and they all had received anti-PDL-1 therapy for one year until the disease progressed. Patient BC17 provided written informed consent to participate in the clinical trial NCT03676985 and the current study. Four patients BC3, BC5, BC6, and BC16 agreed to donate peripheral blood to explore the efficacy of anti-TIGIT therapy in vitro. Detailed clinical characteristics information about patients is provided (Supplementary Table 1).

The study population consisted of 177 patients who presented at 1st Affiliated Hospital of Sun Yat-sen University with primary non-metastatic osteosarcoma from 1999 to 2008. Methotrexate(MTX), cisplatin (DDP), doxorubicin (ADM), ifosfamide (IFO) were using during chemotherapy. All patients received neo-chemotherapy followed by surgical resection and post-operative chemotherapy (Figure 1); "standard" chemotherapy consisting of 4 neo-chemotherapy courses and 8 or more post-operative chemotherapy courses.
Fasting morning blood samples were collected at diagnosis, analyzed for pre-chemotherapy serum ALP (pre-ALP). Because growth influences ALP expression [15 (link)], 150 U/L was considered as the upper normal serum ALP limit in patients less than 18 years, and 110 U/L in those 18 years or older. Pre-ALP values were divided into three categories: normal (below the upper normal limit), high (elevated to less than twice the upper limit), and very high (elevated more than twice the upper limit).

This study was approved by the Ethics Committee of Affiliated Tumor Hospital of Xinjiang Medical University and informed consent was obtained from all patients. Inclusion criteria: primary malignant tumor of bone near the knee joint; neoadjuvant chemotherapy is effective; the tumor did not invade the epiphyseal plate; the tumor did not invade important blood vessels and nerves; no infection. Exclusion criteria: pathological fracture; no limb preservation conditions; tumor invading epiphysis. There were 3 male and 2 female patients, the age range was from 8 to 14 years, with an average of 11.6 years. Distal femoral lesions were observed in 2 cases and proximal tibial lesions in 3 cases. All patients underwent X-ray, computed tomography, magnetic resonance imaging, and emission computed tomography examination, and a biopsy was performed after the examination. All cases were common osteosarcomas with no distant metastasis. According to the Enneking staging system, all cases were classified as stage IIB. The distance between the epiphyseal plate and the tumor was >1 cm in all cases. The magnetic resonance imaging image San Julian classification^{[5 (link)]} was applied to classify the lesions. Type I lesions are defined as a distance from the edge of the tumor to the epiphyseal plate >2 cm; for Type II the distance from the edge of the tumor to the epiphyseal plate <2 cm or adjacent; for Type III the epiphyseal plate is partially in contact with the tumor or invaded epiphysis. All cases were classified as San Julian I or San Julian II and the epiphysis could be preserved. All the patients were treated with preoperative neoadjuvant chemotherapy, surgery, and postoperative adjuvant chemotherapy. All lesions were sensitive to preoperative neoadjuvant chemotherapy, and no pathological fractures occurred during chemotherapy. The chemotherapy regimen consisted of cisplatin 100 mg/m², adriamycin 80 mg/m², methotrexate 12 g/m², and ifosfamide 12 g/m².

We selected 59 cases of OS from the databases of the First Affiliated Hospital of Nanchang University, China, and obtained their corresponding tissue samples. Before the biopsy stage, the patients had not received radiotherapy and preoperative chemotherapy. When OS was diagnosed, all patients received combined chemotherapy, such as high-dose ifosfamide and doxorubicin, cisplatin, and methotrexate, and then underwent a partial removal of the primary tumor, which was followed by an adjuvant chemotherapy cycle. After the completion of chemotherapy, patients were followed up every 3 months for 6 years.

We identified patients who underwent treatment at our hospital from August 2005 to August 2015 using electronic health records, followed by a manual reviewal to assess eligibility. We included patients with pathologically confirmed ENKTL and treated with curative intent radiotherapy. Patients who underwent radiotherapy for palliative intent and those who could not accomplish the planned treatment sequence were excluded. This study was approved by the Institutional Human Research Ethical Committee. Clinical staging of all patients was determined by positron emission tomography-computed tomography according to the Lugano classification criteria [5 (link)].
The patients were treated with radical radiotherapy, with a total dose of 50-60 Gy in 25-30 fractions (1.8-2 Gy per fraction). The radiotherapy techniques utilized were three-dimensional conformal radiotherapy (3D-CRT) and intensity-modulated radiotherapy (IMRT) (Figure 1, Table 1).
The treating physician decided to administer concurrent systemic treatment. The regimen comprising dexamethasone, etoposide, ifosfamide, and carboplatin (DeVIC), was the most frequently employed concurrent regimen with radiotherapy.
Data analyses were performed using the R software package. Statistical significance was set at ˂0.05. PFS was calculated from the date of radiotherapy initiation until disease progression or death from any cause. OS was estimated from the date of radiotherapy initiation to death from any cause. Considering patients lost to follow-up, OS data were censored on the date the patient was last seen alive. Toxicity data were evaluated from electronic medical records and graded retrospectively based on the Common Terminology Criteria for Adverse Events v5.0.