Objective To investigate the clinicopathological features and risk factors of lymph node metastasis of gastrointestinal neuroendocrine neoplasms (GI-NENs).Methods The retrospective case-control study was conducted.The clinicopathological data of 467 patients with GI-NENs who were admitted to the Fourth Hospital of Hebei Medical University from January 2006 to December 2015 were collected.Observation indicators:(1) occurrence sites and pathological classification of GI-NENs;(2) pathological characteristics of surgical specimens of GI-NENs;(3) univariate analysis and multivariate analysis affecting lymph node metastasis of GI-NENs:sex,age,tumor location,tumor diameter,pathological classification,pathological stage and tumor invasive depth.The univariate analysis and multivariate analysis were respectively done using the chi-square test and Logistic regression model.Results (1) Occurrence sites and pathological classification of GI-NENs:of 467 patients with GI-NENs,tumors of 304,15,7,14 and 127 patients were located at stomach,duodenum,small intestine,colon and rectum,respectively.Tumor diameter was 0.3-12.0 cm,with an average diameter of 2.2 cm.Of 467 patients with GI-NENs,G1 and G2 of neuroendocrine tumors (NETs),G3 of neumendocfine carcinomas (NECs) and mixed adenoneuroendocfine carcinomas (MANECs) were respectively detected in 209,64,146 and 48 patients.Lymph node metastasis rate of GI-NENs was 31.48％ (147/467).(2) Pathological characteristics of surgical specimens of GI-NENs:NETs were high-differentiated NENs.Ceils of NETs were solid and nest-,trabeculum-and tubular-shaped,and consisted of small or medium cells,with moderate amount or massive cytoplasms,round or oval nucleus,particle-shaped chromatin,unobvious nucleolus and positive endocrine markers.There were abundant of small blood vessels and surrounding fibrous stroma in peripheral tumor cell nests.NECs were low-differentiated NENs and included small cell carcinoma and large cell NEC.Cells of small cell carcinoma were small round or oval and looked similar to lymphocytes,with few amount cytoplasms,fine granularshaped or hyperchromatic nucleus and common mitosis figures.Cells of large cell NEC were large and greater than 3 lymphocytes,arrayed in organoid-or chrysanthemum-shape,with massive cytoplasms,coarse particle-shaped chromatin,obvious nucleus,clear mitosis figures and large laminar-shaped necrosis.There were different positive expressions of endocrine markers between small cell carcinoma and large cell NEC.MANECs had the characteristics of glandular cavity formation of traditional adenocarcinoma and NENs.Results of immunohistochemical staining in 467 patients showed that Ki-67 of 467 patients was positive;CD56 in 379 of 428 with CD56 test was positive;synaptophysin (Syn) in 416 of 422 with Syn test was positive;cytokeratin (CK) in 354 of 396 with CK test was positive;chromogranin (CgA)in 264 of 388 with CgA test was positive;neuron specific enolase (NSE) in 287 of 346 with NSE test was positive.(3) Univariate analysis and multivariate analysis affecting lymph node metastasis of GI-NENs:results of univariate analysis showed that sex,tumor location,tumor diameter,pathological classification,pathological satge and tumor invasive depth were related factors affecting lymph node metastasis of patients with GI-NENs (X2 =20.654,18.182,26.788,184.709,163.738,195.391,P＜ 0.05).Results of multivariate analysis showed that pathological classification and pathological stage were independent influenced factors affecting lymph node metastasis of patients with GI-NENs (HR =2.129,7.171,95％ confidence interval:1.273-3.561,-2.327-22.098,P＜0.05).Conclusions GI-NENs are mostly located on the stomach and rectum.Results of immunohistochenical staining could help diagnosis of GI-NENs.Pathological classification and pathological stage are independent influenced factors affecting lymph node metastasis of patients with GI-NENs.

Objective To detect specific IgM of Lyme disease indirect ELISA using recombinant outer surface protein C(OpsC)of Borrelia burgdorferi in vitro was established. Methods Coated concentration of recombinant OspC and dilution multiple of serum anol concentration of enzyme secondary antibody were determined by block,and degree of percision.specificity interference and interruption test were performed. Results Best concentration of OspC was 150 μg/L.within-run CV was 6.3% between-run CV was 11.8%.Confimred 33 Lyme patients in clinic and 57 controls were examined meanwhile by this method and import ELISA kit,coincidena rate was 97.8%.Conclusion This ELISA using recombinant OspC was a good method for early diagnostic of Lyme disease.

Objectives To compare the efficacy and adverse effects of combining all-trans retinoic acid and arsenic triox-ide with or without anthracyclines on the treatment of childhood acute promyelocytic leukemia (APL) patients. Methods The retrospective study included 46 children as newly diagnosed APL from January 1st, 2001 to December 31st , 2012. Efficacy and adverse effects for different induction therapies and in high and low white blood cell (WBC) count subgroups were studied. Results In the non antharcycline containing group, 2 patients died during remission induction, and in the antharcycline containing group none of the patients died. No statistical difference was observed between the antharcycline containing group and the non antharcycline containing group in complete remission, the length of time to achieve molecular complete remission and minimal residual disease quantitative analysis at the end of the induction. The mean duration of high WBC count subgroup in the anthar-cycline containing group was shortened than that of the non antharcycline containing group (P<0.05). The recovery time of the abnormal coagulation was found similar between these two groups. Conclusions The use of antharcycline in induction therapy could shorten the duration of high WBC count and reduced the WBC count peak , thus reduces the risk of early death.

Objectives To investigate the inlfuence of polymorphisms of SLC19A1 80G>A, MDR1 exon26C>T and MDR1 exon21G>T/A on curative effect and adverse reaction of high-dose methotrexate in patients with acute lymphoblastic leukemia. Methods MALDI-TOF-MS technique was used to detect the polymorphisms of SLC19A1 80G>A, MDR1 exon 26C>T and MDR1 exon21G>T/A in 108 patients with acute lymphoblastic leukemia (ALL). The relationship of genetic polymorphism, survival rate and toxicity was analyzed. Results The 36-month event-free survival was not related to any polymorphisms of MDR1 and SLC19A1. Patients with mutant types of MDR1 exon26C>T and MDR1 exon21G>T/A showed a much higher MTX plasma levels at 24 hours and higher incidence of hepatic injury (P<0.05). Conclusions The genetic polymorphism of MDR1 exon26>T, MDR1 exon21G>T/A has a large inlfuence on hepatic toxicity and plasma concentra-tions of MTX.

Objective: To analyze the clinical outcome and the prognostic factor in pediatric patients with core binding factor-acute myeloid leukemia (CBF-AML). Methods: A total of 121 newly diagnosed pediatric CBF-AML patients enrolled from Aug. 2005 to Sep. 2017 were retrospectively reviewed. Cumulative incidence of relapse (CIR), event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method and prognostic factors were evaluated by Cox regression with SPSS. Results: Of the 121 patients, 120 patients were assessed for bone marrow remission after induction chemotherapy. 100 cases (83.3%) achieved complete remission (CR) after the first course of chemotherapy. 119 cases (99.2%) achieved CR after the second course of chemotherapy. Of the 121 patients, 13 patients (10.7%) had recurrence with the median interval of recurrence as 13.8 months (3.7 to 58.8 months). 17 patients (14.0%) died. The CIR, EFS and OS at 3 years were 12.7%, 77.5% and 82.8%, respectively. The factors including age at diagnosis, sex, initial WBC count, presence of extramedullary leukemia, C-KIT expression, additional chromosomal abnormalities, and CR after the first course of chemotherapy were analyzed by multivariate regression analysis of Cox. Multivariate analysis identified that additional chromosomal abnormalities was the only independent risk factor affecting OS (HR=4.289, 95%CI 1.070-17.183, P=0.040). Conclusions Pediatric CBF-AML was a unique setting of prognostic subtypes. Chemotherapy produced good responses. Additional chromosomal abnormalities was the only independent risk factor for OS in pediatric CBF-AML.

Objective To explore the prognostic value of quantitative monitoring of RUNX1-RUNX1T1 fusion gene in pediatric t(8;21)/RUNX1-RUNX1T1 positive acute myeloid leukemia(AML).Methods A total of 81 new-ly diagnosed AML children with t(8;21)/RUNX1-RUNX1T1 positive were enrolled in the People′s Hospital,Peking University,between August 2005 and January 2016.RUNX1-RUNX1T1 gene copy number of all the patients was analyzed by real-time quantitative PCR(qPCR)technology at diagnosis and after therapy in all patients.Cumulative incidence of relapse rate(CIR),event-free survival(EFS)rate and overall survival(OS)rate were estimated by Ka-plan-Meier method and prognostic factors were evaluated by COX regression. Results The level of RUNX1-RUNX1T1 gene on diagnosis was used as the baseline to determine whether the level of gene after treatment had a more than 3 logarithmic(3 log)reduction.After 2 courses of induction therapy,the patients with a more than 3 log reduction of RUNX1-RUNX1T1 transcript levels(≥3 log)had better EFS rate(82.4% vs.57.6%,χ2=7.454,P<0.01),and better OS(93.6% vs.59.3%,χ2=9.703,P<0.01),compared to the patients with a less than 3 log reduction(<3 log).Multivariate analysis showed that 3 log reduction in RUNX1-RUNX1T1 transcript levels after 2 courses of in-duction therapy was an independent prognostic factor for EFS rate[hazard ratio(HR)=4.223,95% confidence interval (CI):1.507-11.836,P<0.01]and OS rate(HR=5.002,95%CI:1.282-19.516,P<0.05).When periodically monitoring the RUNX1-RUNX1T1 gene,63 out of the 81 children patients were monitored for more than 6 times.The patients who had a more than 3 log reduction of gene before,but then those whose gene transcript level rose more than 1 log level were divided into group A,and the remaining patients were divided group B,and the difference of CIR was statistically significant between group A and group B(46. 7% vs. 4. 7%,P <0. 01). Conclusions RUNX1-RUNX1T1 gene copy number was detected with qPCR method in pediatric t(8;21)/RUNX1-RUNX1T1 positive AML,which can determine the treatment effect,predict the recurrence of patients and assess long-term prognosis.Thus it has great clinical application value.

Objective To explore the prognostic significance of bone marrow examination at different time points by using different methods during induction therapy.Methods From Feb.2004 to Jan.2013,268 newly diagnosed (B acute lymphoblastic leukemia,B-ALL) pediatric patients in Peking University People's Hospital,were enrolled for the study.In this study,the overall survival (OS) ratio and event-free survival(EFS) ratio of patients with different bone marrow morphology on day 8 and day 15 were analyzed.Based on different cut off value of minimal residual disease (MRD) on day 15 and end-of-induction,the OS ratio and EFS ratio of the higher patients and the lower patients were compared.Results Patients with M1,M2 or M3 marrows on day 8 had no significant difference in OS ratio (P =0.372) or EFS ratio (P =0.393).When it came to day 15,patients with M1,M2 or M3 marrows had no significant difference in OS ratio (P =0.050) or EFS ratio (P =0.324).Patients with MRD ＞ 10.00％ on day 15 had lower OS ratio than those with MRD ≤ 10.00％,and it had significant difference(P =0.022).But there was no significant difference in EFS ratio (P =0.191).As for MRD on the end-of-induction,when using the MRD level of 0.0l ％,0.10％,1.00％ as cut-off values,the lower group of end-of-induction MRD was significantly associated with a higher probability of OS ratio and it had significant differences(P =0.018,0.006,0.002),and it showed the same results at EFS ratio (P =0.002,0.000,0.000).Conclusions The bone marrow morphology on day 8 and day 15 during induction therapy had no prognostic significance.The MRD of day 15 had prognostic significance when using 10.00％ as the cut off value.The critical value of MRD on the end-of-induction MRD should be 0.01％ for the prognosis.

Objective: To investigate the significance of extranodal extension of axillary lymph nodes (ALN-ENE) metastases in post-operative primary invasive breast carcinoma of non-specific type. Methods: Six hundred and thirty-eight invasive breast cancer cases confirmed by postoperative pathological examination were collected from January 2006 to December 2008. The relationship of lymph node metastases and ALN-ENE with other lymph node parameters and patient outcome was analyzed. Results: Among 638 cases, 263 (41.2%) showed axillary lymph node metastases. ALN-ENE was present in 91 cases (36.4%). The rate of ALN-ENE increased with pT stage and tumor size. Five-year recurrence-free survival rate (RFS) and 5-year overall survival rate (OS) was 86.6% and 91.2% respectively for ALN-ENE positive group, and both were lower than ALN-ENE negative group (P<0.01). One hundred and forty-nine patients with 1 to 3 positive lymph nodes had a 5-year RFS of 91.9%, and 5-year OS of 92.3%, less than ALN-ENE negative group (P<0.01). Univariate analysis showed ALN-ENE positively correlated with lymph node metastasis. Multivariate analysis suggested that ENE was associated with increased recurrence risk and shortened recurrence-free and overall survival, especially in patients with 1 to 3 positive lymph nodes; and it was an independent prognostic factor (P<0.01). Conclusions The number of lymph nodes metastases is an important predictor of survival in breast cancer patients. ALN-ENE is an independent risk indicator for recurrence and overall survival. For patients with 1 to 3 metastatic axillary lymph nodes, ALN-ENE could alter the patient′s clinical pathologic staging, and therefore it is an independent prognostic factor.

Objective: To explore the clinical features and prognostic factors of Ph-positive and/or BCR-ABL positive acute lymphoblastic leukemia (Ph⁺ ALL) in children. Methods: The clinical data of 68 Ph⁺ ALL children who were treated at Peking University People's Hospital from December 2006 to December 2016 was retrospectively reviewed. Survival analysis were estimated by Kaplan-Meier method. Univariate analysis was estimated by Log-rank test and Chi-square, and multivariate analysis was estimated by Cox proportional hazards regression model. Results: In the 68 cases, the proportion of male to female was 2.1∶1, with a median age of 8 (1-16) years, and the median overall survival (OS) and disease free survival (DFS) were 16.8 months and 13.5 months, respectively. The early response rate to treatment was 43.9%, with myeloid-antigens-expression group lower than the non-expression group (29.6% vs 61.3%, χ²=5.814, P=0.020); The complete remission (CR) rate after one-course induction therapy was 86.2% (56/65), with good-response group higher than the poor-response group (100.0% vs 74.2%, χ²=6.680, P=0.003);The CR rate after induction in patients receiving imatinib plus chemotherapy was higher than the patients receiving chemotherapy only (94.9% vs 73.1%, χ²=5.185, P=0.024). The 2-and 5-year OS were (61.4±7.0)% and (50.8±8.1)%, respectively. The 2-and 5-year DFS were (54.6±6.8)% and (48.6±7.3)%, respectively. Univariate analysis showed that the initial WBC, LDH, spleen size, liver size, with-myeloid-antigens-expression, early response to treatment, MRD (BCR-ABL) after one-course induction, application of imatinib and different treatment options affected 2-year OS rate (all P<0.05). LDH, spleen size, liver size, with-myeloid-antigens-expression, early response to treatment, MRD (BCR-ABL) after one-course induction, application of imatinib and different treatment options affected 2-year DFS rate (all P<0.05). Multivariate prognostic analysis for OS (RR=45.7, 95% CI 1.4-1 528.2, P=0.033) and DFS (RR=52.3, 95% CI 1.6-1 725.9, P=0.026) showed that the spleen ≥ 3 cm was the independent risk factor. Conclusions Pediatric Ph⁺ ALL is a special condition with unique clinical and biological features. The early response to treatment was poor in patients with myeloid-antigens-expression, which resulted in a low CR rate after one-course induction and the administration of imatinib can remarkably improve the CR rate. Initial spleen ≥ 3 cm is an independent prognostic factor. The efficacy of chemotherapy alone is poor, and imatinib combined with chemotherapy is applauded in the aim of improving outcomes.

Objective:To explore the molecular response and prognostic factors of pediatric patients with Ph-positive acute lymphoblastic leukemia (Ph + ALL) treated by tyrosine kinase inhibitors (TKI) with chemotherapy in TKI era. Methods:The clinical data of children newly diagnosed with Ph + ALL admitted at Department of Pediatrics, Peking University People′s Hospital from August 2006 to February 2017 were retrospectively reviewed.The molecular biological characteristics and survival prognosis of the 30 patients who received continuous TKI with chemotherapy from early induction combined and no subsequent transplantation were analyzed. Results:The 30 patients with Ph + ALL had 19 males and 11 females with a median age of 8-year-old (ranging from 2 to 16 years). The complete remission (CR) rate after the first cycle of induction chemotherapy was 96.7% (29/30 cases), with overall CR rate of 100.0%; Before treatment, the mean level of BCR/ ABL mRNA in the 30 patients was 73.2% (0.12%-160.60%) and the level declined significantly with the progression of chemotherapy courses, reaching the plateau stage at the 6 th month of chemotherapy ( Z=-1.922, P>0.05); nine patients had recurrence, with a median recurrence time of 7 months (3.7-58.8 months). Univariate analysis showed that age ( P<0.05), the lever of minimal residual disease (MRD) after induction chemotherapy ( P<0.01) and the MRD level at the 3 th month of induction chemotherapy ( P<0.01) affected the recurrence rate.The median follow-up time of 30 patients was 42.6 months (6.4-96.5 months), and the 3-year overall survival (OS) rate and event-free survival (EFS) rate were (78.6±7.8)% and (72.4±8.4)%, respectively; Cox multivariate analysis showed that the initial white blood cell count ≥34.0×10 9/L ( OR=11.955, 95% CI: 1.075-132.899, P<0.05) and BCR/ ABL mRNA reduction less than 3 log from baseline [major molecular response (MMR)] at the 3 th month of induction chemotherapy ( OR=8.563, 95% CI: 1.254-58.478, P<0.05) were independent risk factors affecting the 3-year EFS rate.In addition, the initial white blood cell count ≥34.0×10 9/L ( OR=14.327, 95% CI: 1.843-243.592, P<0.05) was also an independent risk factor affecting the 3-year OS rate. Conclusions:The application of TKI can significantly deepen the molecular response of Ph + ALL in children.In the TKI era, the initial white blood cell count ≥ 34.0×10 9/L and BCR/ ABL mRNA reduction less than 3 log from baseline (MMR) at the 3 th month of induction chemotherapy are independent risk factors for the long-term survival of pediatric Ph + ALL.