Aim To amplify P30 gene and express P30 fusion with GST Methods P30 gene was smplified from T. gondii chromosomal DNA and ligated to pGEM-T and pGEX-4T-1. Screening-positive recombinants were induced for expression, which was subsequently detected by WB Results P30 gene was amplified and GST-fusion was confirmed by rabbit antiT. gondii serum. Conclusions The construction of pGEM-T-P30 and pGEX-4T-1-P30, together with the recombinant protein would lay a base for further investigation of P30 at a molecule-level and application to diagnosis and vaccination

Objective To discuss the diagnostic value of serum procalcitonin (PCT) and C-reactive protein (CRP) in children suffering from malignant solid tumor with concurrent infection and in tumor progressing stage.Methods A total of 94 cases of malignant solid tumor at Department of Pediatrics,Beijing Tongren Hospital,Capital Medical University from March to August 2016,were collected and divided into 2 groups:infection group (n =17) and non-infection group (control group,n =77).Meanwhile,the control group were further divided into stable tumor group (n =48) and the advanced tumor group (n =29).All of the serum specimens were detected for PCT and CRP levels.PCT and CRP levels were compared and statistically analyzed among different groups.The median(four digit spacing) was used to describe the data.Results PCT level significantly increased in the infection group [0.67 (1.35) μg/L] compared with that of the control group [0.09 (0.08) μg/L],and the difference was statistically significant (Z =-6.371,P =0.000).CRP level significantly increased in the infection group[95.00(93.42) mg/L] compared with that of the control group[1.99 (12.49) mg/L],and the difference was statistically significant (Z =-5.555,P =0.000).During diagnosing infection,the area under the receiver operating characteristic curve (ROC curve) of PCT was 0.995(95％ CI:0.987-1.000),and the area under the ROC curve of CRP was 0.932(95％ CI:0.876-0.988);when the cutoff value of PCT was 0.468 μg/L,the sensitivity was 76.5％,and the specificity was 100.0％;when the cutoff value of CRP was 10.17 mg/L,the sensitivity was 88.2％,and the specificity was 72.7％.Compared to stable tumor group,PCT level of advanced tumor group significantly increased[0.12 (0.08) μg/L vs.0.07 (0.06) μg/L],and the difference was statistically significant(Z =-2.886,P =0.004);CRP level of the advanced tumor group sig nificantly increased [10.34 (27.84) mg/L vs.1.02 (2.12) mg/L],and the difference was statistically significant (Z =-4.862,P =0.000).During diagnosing tumor progression,the area under the ROC curve of PCT was 0.697 (95 ％ CI:0.572-0.823),and the area under the ROC curve of CRP was 0.832(95％ CI:0.738-0.926);when the cutoff value of PCT was 0.094 μg/L,the sensitivity was 72.4％,and the specificity was 66.7％;when the cutoff value of CRP was 2.39 mg/L,the sensitivity was 72.4％,the specificity was 72.9％.Correlation analysis showed that the PCT and CRP levels were positively associated with tumor progression,and the correlation of CRP (r =0.558,P =0.000) was much higher than that of PCT (r =0.331,P =0.003).Conclusions Both serum PCT and CRP are effective indicators in diagnosing tumor with concurrent infection.PCT is better in diagnosing infection than CRP,and is less influenced by tumor.CRP is a better way to identify whether tumor was in progress than PCT.Joint detection of 2 indexes in identification of children suffering from malignant solid tumor with concurrent infection and in progress has practical value for the diagnosis and treatment.

Objective:To explore the cellular protective effect and adverse reactions of amifostine in the chemotherapy of malignant solid tumor in children.Methods:A total of 62 children with malignant solid tumors receiving 253 times of chemotherapy who were admitted to the Pediatrics Single Center of Beijing Tongren Hospital from April 2018 to April 2020 were selected and divided into the experimental group (amifostine was used before chemotherapy, 113 times in total) and the control group (amifostine was not used before chemotherapy, 140 times in total) according to stratified random sampling. The self-control method was used to compare the therapeutic effects and adverse effects of the use of amifostine or not in the same child under the same chemotherapy regimen.Results:Compared with the control group, the duration of agranulocytosis [(6.7±3.0) d vs. (9.5±4.3) d, t = 3.788, P < 0.05], the duration of platelet reduction (<20×10 9/L) [(3.6±1.3) d vs. (5.4±3.2) d, t = 2.037, P < 0.05], the time of receiving recombinant human granulocyte colony-stimulating factor (rhG-CSF) treatment [(6.5±3.5) d vs. (10.0±2.8) d, t = 3.049, P < 0.05] and the time of antibiotic treatment during infection [(5.0±2.5) d vs. (8.2±2.5) d, t = 3.558, P < 0.05] in the experimental group were all shorter; the amount of platelet input required [(0.7±0.5) U vs. (1.5±0.8) U, t = 2.873, P < 0.05] was less than that of the control group. Oral mucosal ulceration occurred in only 4 (3.5%) times in the experimental group, which was lower than that in the control group [12 (8.6%) times] ( χ2 = 4.634, P = 0.033). Regardless of the cost of amifostine itself, there was a statistically significant difference in treatment cost between the experimental group and the control group ( P = 0.034), and the length of hospital stay in experimental group was relatively short ( P = 0.012). The patients were more prone to nausea and vomiting and hypocalcemia when treated with amifostine. Conclusions:Amifostine can effectively protect normal tissue cells in chemotherapy of children with malignant solid tumor and its adverse reactions are mild.

Objective To summarize the clinical,pathological and radiological features of retinoblastoma (RB)with central nervous system (CNS)metastasis.Methods Twenty -three patients were confirmed to have RB with CNS metastasis in Beijing Tongren Hospital from December 2005 to December 201 3,and their clinical data were retrospectively analyzed.Results (1 )The incidence of RB with CNS metastasis was 1 .83% (23 /1 260 cases),and the incidence of CNS metastasis was 7.64%(1 1 /1 44 cases)if RB with optic nerve involvement which was confirmed through pathology.At first visit 1 0 cases had a confirmed diagnosis of RB with CNS metastasis,while the other 1 3 cases had a diagnosis of RB in extraocular stages.(2)Eleven patients with RB in extraocular stages had the pathological evi-dence of optic nerve involvement which was pathologically confirmed,6 patients had optic nerve involvement with cho-roid involvement,2 patients had optic nerve involvement with nerve sheath involvement.(3)All the patients whose diagnosis of CNS metastasis had the radiological evidence of CNS metastases,and 1 case had autopsy and pathological evidence for extensive brain metastases.The main radiological feature of CNS metastasis was meninges metastasis.Thir-teen cases had the radiological features of meninges metastasis and 5 cases accompanied with spinal cord meninges me-tastasis.Then,the following radiological feature was that 7 cases had the mass of suprasellar pool.(4)Twelve cases with CNS metastasis showed the cerebrospinal fluid (CSF)RB cells positive,and the positive rate was 52.1 7%(1 2 /23 ca-ses).Conclusions (1 )The occurrence of RB with CNS metastasis is very low,but the risk of CNS metastasis in-creased in the patients with the pathological evidence of the optic nerve involvement.(2)The main site of CNS metasta-ses is meninges,followed by mass formation in the suprasellar pool.The main route for CNS metastases was along the optic nerve and /or nerve sheath directly invading and /or disseminating via the CSF.(3)The positive rate of CSF cy-tology is low,and it can be improved through repeated examination.Enhanced magnetic resonance imaging should be the most effective radiology examination for diagnosis and monitoring CNS metastasis.

Objective To explore the treatment and prognosis of advanced stage childhood hepatoblastoma with pulmonary metastasis.Methods Fifty-six cases of advanced stage hepatoblastoma with pulmonary metastasis diagnosed through pathology from April 2006 to June 2014 in Department of Pediatrics,Beijing Tongren Hospital Affilia-ted to Capital Medical University were enrolled,among them 33 cases were males and 23 cases were females,and the median age was 2.33 years old(1 month-15 years and 1 month old).The clinical effects of multidisciplinary therapy were analyzed.Results (1)Follow-up studies were conducted till December 2016,in which 21 cases of 56 children achieved complete remission,the complete remission rate was 37.5%(21/56 cases),while 12 cases were partial re-mission and 14 cases were deceased,and the effective rate reached 58.9%(33/56 cases).The follow-up period of 41 children were over 24 months,in which the 2-year free event survival(EFS)rate was 37.5%,2-year overall survi-val(OS)rate was 75.0%,5-year survival rate was 42.4%,and the 95% average survival confidence interval was 35.7-55.9 months.(2)The OS rate of children with small age(≤3 years old)was 88.1%(36/42 cases),the ove-rall prognosis was better than that of >3 years old children(35.7%,5/14 cases)(P=0.003).The survival rate of children with complete tumor resection[OS rate was 89.2%(33/37 cases)]was significantly higher than that of the incomplete excision[OS rate was 47.4%(9/19 cases)],and the difference was statistically significant(P=0.001). The prognosis of epithelial type cases was better than that of other types,and the difference was statistically significant (P<0.05),while the fetal type prognosis was the best,and the difference was statistically significant(χ2=8.56,P=0.014).The growth of alpha fetoprotein was negatively correlated with the clinical efficacy and prognosis(r=-0.468, P=0.023).Conclusions Lung is the most common metastatic site of hepatoblastoma,and the marginal lung metasta-sis is more common.With insidious onset and poor prognosis.Therefore,it should be treated with early diagnosis and multidisciplinary therapy to improve prognosis.

Objective To analysis the prognosis of head and neck rhabdomyosarcoma (RMS) in pediatrics. Methods The clinical data of 98 pediatric patients with head and neck RMS admitted from February 2005 to September 2017 were retrospectively analyzed. The clinical staging and risk grouping were performed according to international rhabdomyosarcoma study group criterion. The treatment included chemotherapy, radiotherapy and surgery accordingly. The association of prognosis with the disease characters, treatment strategy of 98 patients RMS was analyzed by Chi-square test and Kaplan-Merier analysis with SPSS 20.0 software. Results There were 53 boys (54.1％) and 45 girls (45.9％) with a median age of 70 months (2-161 months). There were 43 cases of orbit RMS(43.9％), 46 cases of paramenigneal RMS (46.9％)and 9 cases of non-paramenigneal and non-orbit RMS(9.2％). Twenty seven patients were classified as low risk (27.6％), 42 patients as medium risk (42.9％),and 29 patients as high risk (29.5％). The patients were followed up for a medium of 42 months (9-149 months), 37 cases were dead, 60 cases were survival, and 1 lost the follow-up. Kaplan-Merier survival analysis showed that the overall survival (OS) was 61.9％(60/97), and the average survival time was (92.3±6.9) months. Univariate analysis showed that the OS of patients with paramenigneal RMS was poorer than that of patients with orbit RMS, non-paramenigneal and non-orbit RMS. The OS of high risk RMS patients was poorer than that of intermediate and low risk patients (χ2=6.132, P=0.047). Conclusions The prognosis of head and neck RMS in children is generally good, but the prognosis of paramenigneal RMS and high-risk RMS is significantly declined. The primary site of paramenigneal and high risk are independent risk factors for the prognosis of head and neck RMS in children.

Objective: To study on the mRNA expression level of IN1 gene associated with the clinical efficacy and prognosis of hepatoblastoma(HB) in children and to elucidate the early warning value of IN1 gene for prognosis and chemotherapy sensitivity. Methods: Forty HB patients were selected based on primary diagnosis and treated from January 2015 to May 2018 in Tongren Hospital Affiliated to Capital Medical University. The expression level of INI1 gene was detected by using fluorescence immuno-PCR. Clinical data of children with HB were collected, including staging, grouping, risk, efficacy and prognosis of chemotherapy. The relationship between the mRNA expression of IN1 gene and clinical data was analyzed by SPSS 21.0 software. Results: (1)Clinical characters: the medium age of 40 HB patients was(30.50±2.39) months, male 29 cases (72.5%), and female 11 cases (27.5%). There was 1 case (2.5%) of stage Ⅱ HB, 13 cases (32.5%) of stage Ⅲ HB, and 26 cases (65.0%) of stage Ⅳ. The low-and intermediate- risk group had 12 cases (30.0%) of HB, and the high-risk group had 28 cases (70.0%) of HB.(2)Clinical efficacy and prognosis: by July 1^st, 2018, the medium following-up time was (12.2±10.1) months, 4 patients (10%) were treated by chemotherapy without surgery, and 36 patients (90%) were treated by surgery .All patients (100%) received chemotherapy.The average cycle of chemotherapy lasted(13.17±0.02). Ten patients were dead and 30 patients survived till following-up.The overall survival (OS) rates were 75% and the event-free survival rates were 50% (20 patients with HB). (3) IN1 gene mRNA expression: The average PCR quantitative of IN1 gene mRNA of 40 HB patients(2^-ΔCT)was 0.31±0.70.The average mRNA quantitative of HB patients (2^-ΔCT)with high-risk and low/intermediate-risk group was 0.23±0.43 and 0.48±1.13(t=6.363, P=0.05). According to histology diagnosis, the average mRNA quantitative of small cell undifferentiated (SCU) type (12 cases)and other type (25 cases) was 0.09±0.11 and 0.43±0.86(t=4.533, P=0.04). The average mRNA quantitative of patients with or without radical surgery was 0.04±0.03 and 0.34±0.74 (t=2.935, P=0.022). The data of children with poor chemotherapy sensitivity(19 cases)and those sensitive to chemotherapy (21 cases) were 0.03±0.04 and 0.30±0.82, and the difference was statistically significant (t=5.688, P=0.018). Conclusions Poor therapeutic effect results in low mRNA expression of IN1 gene in children of HB.The IN1 gene expression could be an early warning factor for treatment sensitivity and prognosis.

Objective:To investigate the clinical features, diagnosis, treatment and prognosis of advanced clear cell sarcoma of kidney(CCSK) in children.Methods:The clinical data of 10 children with advanced CCSK hospitalized in Department of Pediatrics of Beijing Tongren Hospital, Capital Medical University from January 2014 to December 2017 were collected, and their clinical features, diagnosis, treatment and prognosis were analyzed retrospectively.Results:(1) Clinical features of CCSK: ten cases of CCSK included 6 boys and 4 girls, with the median onset age of 32 months; 7 cases were left CCSK and 3 cases were right CCSK.There were 9 cases of stage Ⅲ and 1 case of stage Ⅳ at the time of initial diagnosis, when 4 cases were misdiagnosed as other renal tumors at the time of initial diagnosis(40%, 4/10 cases). Five patients with stage Ⅲ CCSK had recurrence and metastasis during treatment and follow-up, and the main distant metastasis sites were lung, bone, liver and brain.(2) Treatment and prognosis of CCSK: seven cases received surgery combined with radiotherapy and chemotherapy, and 3 cases whose parents gave up treatment adopted non-standardized treatment.The median follow-up time was 33.5 months.Seven patients survived and 3 cases died.The 3-year overall survival rate of all 10 patients was 65.6%.The 3-year overall survival rate of stage Ⅲ was 74.1%, and that of stage Ⅳ was 0.The prognosis of stage Ⅲ was significantly better than that of stage Ⅳ( χ2=9, P=0.003). Among the 5 recurrent cases, only 1 case achieved completely remission, 2 cases achieved partially remission, 1 case suffered from disease progression and 1 case died.The 3 cases without recurrence were given standardized treatment of surgery, che-motherapy and radiotherapy, and all were completely remitted. Conclusions:CCSK is easy to be misdiagnosed, and the risk of recurrence and distant metastasis is high in stage Ⅲ patients during treatment and follow-up.Stage Ⅲ patients who actively receive standard treatment including surgery, chemotherapy and radiotherapy have good prognosis, while the mortality of patients with relapse and distant metastasis is high.