Objective To analyze the effects of curcumin combined with cisplatin on the growth and lymphatic metastasis of cervical cancer xenografts in nude mice.Methods The human cervical carcinoma Caski cells xenotransplanted tumor models were established.Inoculated mice were randomly divided into 4 groups according to random number table:control (normal saline 10 ml/kg),Cur (curcumin 100 mg/kg),Cis (cisplatin 3 mg/kg) and Cur + Cis group (100 mg/kg curcumin +3 mg/kg cisplatin).The tumor volume and anti-tumor rate were calculated.The mRNA levels of macrophage migration inhibitory factor (MIF) and vascular endothelial growth factor-C (VEGF-C) were analyzed by real-time fluorescent quantitative polymerase chain reaction (PCR).Results The inhibitory rates of Cur group,Cis group and Cur + Cis group were 40.8％,53.3％ and 60.0％.After 15 days treatment,the tumor volumes of the control group,Cur group,Cis group and Cur + Cis group were (123.44 ± 35.62),(71.72 ± 28.36),(65.47 ± 18.32),(53.44 ± 10.79) mm3.The growth rates of tumor volume in Cur group,Cis group and Cur + Cis group were slower,Cur + Cis group could more effectively inhibit tumor volume growth.The difference among the four groups had statistically significance (F =16.890,P =0.000).Real-time fluorescent quantitative PCR detection showed that compared with the control group (1.000),the MIF mRNA expressions in Cur group,Cis group and Cur + Cis group were 0.322 ±0.094,0.154 ± 0.006 and 0.136 ± 0.007,VEGF-C mRNA expressions in the three groups were 0.312 ± 0.068,0.263 ± 0.072 and 0.221 ± 0.041.The differences among groups had statistically significance (F =220.279,P =0.000;F =143.250,P =0.000).MIF mRNA was positively related with VEGF-C mRNA (r =0.815,P =0.001).Conclusion Curcumin combined with cisplatin can inhibit the growth of Carski cell xenografts in nude mice.Through the down-regulating expression of MIF mRNA and VEGF-C mRNA,cisplatin and curcumin can inhibit the lymphatic metastasis of cervical cancer,and it may be one of the important mechanisms of its anti-tumor effects.

Objective To investigate the expression and clinical significance of Smac and HtrA2 in children with acute leukemia(AL).Methods Bone marrow samples were obtained from 77 children with AL (including 32 newly diagnosed children,33 complete remission children and 12 relapsed children)and the control group of 15 children without malignant blood disease.The expressions of Smac and HtrA2 protein were measured by streptavidin/peroxidase immunoperoxidase technique(SP) in all children.SPSS 13.0 software was applied to analyze the statistical data.Results Protein Smac was detected only in some samples,but HtrA2 was detected in all samples.The levels of Smac and HtrA2 protein in newly diagnosed AL children were both higher than those of the complete remission children (x2 =17.38,F =2.36,all P ＜ 0.05) and normal controls (x2 =12.89,F =5.26,all P ＜ 0.05),there was a statistical significance,but compared with those in the relapsed children,the difference had no statistical significance (x2 =1.18,F =1.57,all P ＞ 0.05).The levels of Smac and HtrA2 protein in complete remission children were both higher than those of the normal controls,and the difference had no statistical sigmficance(x2 =1.20,F =2.23,all P ＞ 0.05).In the newly diagnosed children,the levels of Smac and HtrA2 protein in children with acute lymphocytic leukemia(ALL) were higher than those of the acute myeloid leukemia(AML),but the differences had no statistical significance(x2 =0.113,t =1.024,all P ＞ 0.05).In newly diagnosed AL children,the complete remission(CR) rate of the negative expression of Smac(Smac-,90.9％) and the low expression of HtrA2(HtrA2low,84.6％) in the level of protein were higher than those of the positive expression of Smac(Smac +,47.6％) and the high expression of HtrA2 (HtrA2high,47.4％),and there was statistical significance respectively(x2 =5.772,4.596,all P ＜ 0.05).The CR rate of Smac-HtrA2low group (100％) was higher than that of Smac+ HtrA2high group(30.8％)in the children with AL,and the statistical data were of great significance(x =9.692,P ＜0.01).The protein level of Smac in newly diagnosed AL children was correlatedwith HtrA2 (r =0.979,P ＜ 0.001).Conclusions Pro-apoptotic protein Smac and HtrA2 may be involved in and af-fected each other in the pathogenesis and progression in AL,but levels of Smac and HtrA2 protein may be not correlatedwith the types of AL.In newly diagnosed AL children,the high expression of protein Smac and HtrA2 predicts poorprognosis.

Objective To detect the levels of insulin-like growth factors in children with acute leukemia (AL). Methods A total of 50 previously untreated AL patients were selected, meanwhile 30 healthy children were selected as normal controls. AL children were given regular chemotherapy. All cases were not given the brain radiotherapy. The levels of insulin-like growth factor-1 (IGF-1), free insulin-like growth factor-1 (fIGF-1), insulin-like growth factor binding protein 3 (IGFBP-3) in AL patients before treatment and 6 months after complete remission were measured by enzyme-linked immunosorbent assay (ELISA), and were compared with those in normal controls. Results Before treatment, compared with normal controls, the serum levels of IGF-1, IGFBP-3 in AL patients were lower while the level of fIGF-1 was higher, and the differences were signiifcant (P<0.01). At six months after complete remission, the levels of IGF-1 and fIGF-1 in AL patients were similar to those before treatment, but were signiifcantly different from those in control group (P<0.05);the level of IGFBP-3 was signiifcantly higher than that before treatment (P<0.01), but was similar to that in control group. Before treatment, the level of IGFBP-3 in AL patients was positively correlated with the level of IGF-1 (r=0.777, P<0.01), and negatively correlated with the level of fIGF-1 (r=-0.714, P<0.01). Conclusion Insuline-like growth factors were involved in the pathophysiological process in children with AL.

Objective To analyze the functional changes and the clinical significance of B cell specific mono-clonal murine leukemia virus integration site -1 (Bmi -1 )and Th1 /Th2 cells in children with newly diagnosed im-mune thrombocytopenia(ITP)by testing the mRNA expressions of Bmi -1,helper T cell -related cytokine interferon (IFN)-γand interleukin(IL)-4 in children with newly diagnosed ITP.Methods Thirty -six cases of patients with newly diagnosed ITP in the experimental group came from the inpatient and outpatient children admitted to the Depart-ment of Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from April to December 201 3.In the control group,26 cases of children requiring selective operation were admitted to the Department of Pediatric Surgery during the same period.The mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes were detected by means of the reverse transcription -polymerase chain reaction(RT -PCR)method,and were analyzed and compared by t test and linear correlation analysis.Results (1 )The mRNA expressions of Bmi -1,IFN -γand IL -4 in peripheral blood lymphocytes in the experimental group were 2.63 ±0.54,3.84 ±0.43 and 1 .44 ±0.39,respec-tively;while the mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes in the control group were 3.91 ±0.92,2.88 ±0.57 and 1 .87 ±0.34,respectively.The levels of IFN -γof the experimental group were significantly higher than those of the control group (P <0.001 )and the levels of Bmi -1 and IL -4 in the experi-mental group were significantly lower than those in the control group (all P <0.001 ).(2)The mRNA expressions be-tween IFN -γand IL -4 in the peripheral blood lymphocytes in the experimental group were in negative correlation (r =-0.667,P <0.001 ).The mRNA expressions between IL -4 and Bmi -1 in the same group were in a positive correlation (r =0.776,P <0.001 ).There were no correlation in the mRNA expressions between IFN -γand Bmi -1 (r =-0.206,P >0.05).Conclusions Bmi -1 may be involved in the pathogenesis of ITP by regulating Th cell, and Th cell dysfunction may occur in the children with ITP,and the disproportion between Th1 and Th2 may be due to the advantages of Th1 .

Objective To discuss DNA methylation's effect on pathogenesis of pediatric immune thrombocytopenia (ITP)through detecting the expression level of DNA methyltransferases (DNMTs)mRNA in peripheral blood lymphocytes of children with ITP.Methods Two mL peripheral blood was collected from each of 25 children with persistent and chronic ITP and 20 healthy children (the healthy control group)by using aseptic method in the pediatric ward of the First Affiliated Hospital of Xinxiang Medical University from January 2014 to January 2015.First ethylene diamine tetraacetic acid (EDTA) was used as the anticoagulant.Then separate the mononuclear cells,extract RNA and detect expression levels of DNMT1,DNMT3A and DNMT3B mRNA using reverse transcription-polymerase chain reaction (RT-PCR) method.Results (1) The blood platelet (PLT) of children with persistent and chronic ITP was (36.2 ± 19.6) × 109/L,which was obviously lower than the healthy control group(168.8 ±46.8) × 109/L(t =-11.85,P =0.000).(2)The DNMT1 mRNA expression level of children with persistent and chronic ITP was 0.17 ± 0.05,which was obviously lower than the healthy control group (0.27 ± 0.10) (t =-3.912,P =0.001).The DNMT3A mRNA expression level of children with persistent and chronic ITP was 0.20 ± 0.10,which was obviously lower than the healthy control group (0.32 ±0.11) (t =-3.779,P =0.000).The DNMT3B mRNA expression level of children with persistent and chronic ITP was 0.16 ± 0.1 1,which was obviously lower than the healthy control group (0.31 ±0.11) (t =-4.641,P =0.000).(3) There was positive correlation between the expression of DNMT1 and DNMT3B mRNA(r =0.433,P =0.031).There was positive correlation between the expression of DNMT3A and DNMT3B mRNA(r =0.721,P =0.000).Conclusions (1) Children with persistent and chronic ITP have lower expression levels of DNMT1,DNMT3A,DNMT3 B mRNA,which indicates that DNA methylation contributes to the pathogenesis of pediatric persistent and chronic ITP.(2) DNMTs have synergistic effect on DNA methylation of pediatric persistent and chronic ITP.

Objective To study the relationship between DNA methylation and pathogenesis of childhood immune thrombocytopenic purpura (ITP) by examining the expression of DNA methyltransferase 1(Dnmt1) and DNA methyltransferase 3a (Dnmt3a) mRNA in peripheral blood lymphocytes of the children with ITP. Methods Expression of Dnmt 1 and Dnmt3a mRNA in the peripheral blood lymphocytes in 36 children with newly diagnosed ITP and 26 healthy children were detected using RT-PCR. Results Dnmt1 mRNA expression in peripheral blood lymphocytes in children diagnosed with ITP was 3.02±0.49, significantly lower than 4.58±0.52 in the control group (t=11.95, P<0.001). Dnmt3a mRNA expression in peripheral blood lymphocytes in children diagnosed with ITP was 1.49±0.44, signiifcantly lower than 2.41±0.32 in the control group (t=9.12, P<0.001). Conclusions Children with newly diagnosed ITP have lower DNA methylation status in peripheral blood lymphocytes as compared to that in healthy children. The DNA methylation may play an important role in the etiology of acute ITP in children.

Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

Objective:To analyze risk factors for duration of small or medium-sized coronary artery aneurysms (CAA) in children with Kawasaki disease (KD) so as to provide clinical guidance for early and full course treatment.Methods:The clinical data of 68 children diagnosed with KD in the Department of Pediatrics, the First Affiliated Hospital of Xinxiang Medical University from January 2018 to January 2021 were retrospectively analyzed.According to duration of CAA, all cases were divided into 2 groups, duration of CAA ≥ 8 weeks group and duration of CAA <8 weeks group.Risk factors associated with CAA duration were screened using univariate analysis, and then independent risk factors for CAA duration in children with KD were analysed using multiple Logistic regression analysis. Results:A total of 68 cases were enrolled in this study.Among these cases, 45 cases (66.18%) were male and 23 cases (33.82%) were female.The onset age was from 3 months to 10 years old, and the median onset age was 1.59 (1.02-3.19). There were 31 cases in the group with CAA duration ≥8 weeks and 37 cases in the group with CAA duration <8 weeks.Univariate analysis showed that patients with the total fever course >10 days[45.16%(14/31 cases) vs.21.62%(8/37 cases)], time of treatment with intravenous immunoglobulin (IVIG)>10 days[54.84%(17/31 cases) vs.16.22%(6/37 cases)], platelet (PLT)>600×10 9/L[32.26%(10/31 cases) vs.10.81%(4/37 cases)], hypersensitive C-reactive protein (HsCRP) >100 mg/L[38.71%(12/31 cases) vs.13.51%(5/37 cases)] (all P<0.05 ) in the group with CAA duration ≥8 weeks were significantly more than those in the group with CAA duration <8 weeks.However, there were no significant differences in gender, age, type of KD, etiology evidence, hormone application, duration of fever before IVIG application, IVIG sensitivity, IVIG application way, urine leukocytes, white blood cells, hemoglobin, percent of neutrophilic granulocyte, erythrocyte sedimentation rate, glutamic-pyruvic transaminase between the 2 groups (all P>0.05). Multivariate Logistic regression analysis showed that the course of IVIG before application >10 days ( OR=6.589, 95% CI: 1.678-25.867, P=0.007)and HsCRP >100 mg/L ( OR=7.949, 95% CI: 1.947-32.461, P=0.004)were independent risk factors for predicting the duration of KD complicated with small and medium-sized CAA ≥8 weeks. Conclusions:The course of IVIG before application >10 days and HsCRP>100 mg/L are independent risk factors for KD complicated with small and medium-sized CAA lasting ≥8 weeks.

Objectives:To analyze the clinical features of juvenile myelomonocytic leukemia (JMML) and investigate the characteristics of diagnosis and treatment of this disease.Methods:The clinical data of seven children patients with JMML who received treatment in The First Affiliated Hospital of Xinxiang Medical University between April 2015 and February 2020 were retrospectively analyzed. The clinical efficacy of different treatments was analyzed.Results:The median age at diagnosis of JMML was 8 months and 4 days for seven children patients. Fever was the principal cause of treatment, and it was mostly accompanied by hepatosplenomegaly. The median value of peripheral blood leukocyte count was 36.1 × 10 9/L, and it was 4.5 × 10 9/L for mononuclear cell count, 88 g/L for hemoglobin level, and 47 × 10 9/L for platelet count. Myeloid immature cells were found in peripheral blood smears of six patients. Chromosome examination results revealed 7-monomer in one patient, and normal karyotype in six patients. Hemoglobin level was increased in six patients. Gene detection results revealed PTPN11+NF1 mutation in one patient, N-RAS mutation in two patients, and K-RAS mutation in one patient. Three patients gave up treatment, three patients received low-intensity chemotherapy , and these six patients died of complicated infection. One patient received allogeneic hematopoietic stem cell transplantation and the patient survived without any event after 14 months of follow-up. Conclusion:The age of JMML onset is low. JMML has poor clinical specificity. Gene detection is helpful for early diagnosis of JMML. Low-intensity chemotherapy can prolong survival period, but it can not improve prognosis. Infection is the principal cause of death in patients with JMML. Hematopoietic stem cell transplantation is the only possible method to cure the disease.

Objective To study the distribution and clinical significance of allergens in children with adenoid hypertrophy.Methods The allergen detection results of 223 children with adenoid hypertrophy admitted to the Department of Otolaryngology,the Wuxi Second People's Hospital from January 2021 to October 2022 were retrospectively analyzed,while 43 children without adenoid hypertrophy were set as the control group.The allergy test results were compared between the two groups for difference.Results The positive rate of total allergens in adenoid hypertrophy group was 54.7%.The positive rate of total allergens in the control group was 27.9%.The difference was statistically significant(P<0.05).In the adenoid hypertrophy group,the positive rate of inhaled allergens was 41.3%,mainly mites/dusts and molds.The positive rate of ingested allergens was 32.3%,mainly milk and egg white.The adenoid hypertrophy group was divided according to age.The positive rate of total allergens was 54.0%in the 2-7 year old group.The positive rate of total allergens in the 8-14 year old group was 55.7%.There was no significant difference between the two groups(P>0.05).Conclusion Allergen stimulation is one of the causes of adenoid hypertrophy.Mite,mold,milk and egg white are common allergens in children with adenoid hypertrophy in Wuxi.The effect of allergen stimulation on adenoid hypertrophy did not decrease with age.Therefore,the detection of serum allergens can help to understand the specific allergens of children and provide direction for the non-surgical treatment of children with adenoid hypertrophy.