Hepatoblastoma (HB)is the most common malignant liver tumor in children.Surgical resection was the major treatment approach to HB,whereas more than 60% patients had unresectable mass at diagnosis,even though complete resection,there were about 30% patients recurrent.With the introduction of adjuvant chemotherapy,the treat-ment of HB has been developed to multidisciplinary cooperation from single surgical resection,such strategy greatly im-proved the prognosis of children with HB,but there still be a great challenge in the treatment of high -risk patients.All famous children′s HB study groups have begun to focus on the optimization of chemotherapy,surgery opportunity,strate-gy based on risk -stratification,looking for new potential therapeutic targets and prognosis -related risk factors.

Objective:To investigate the clinical value of individualized therapy based on the molecular target detection to improve the prognosis of children with refractory or relapsed malignant solid tumor.Methods:The clinical data of children who were diagnosed with malignant solid tumors between September 1 st 2012 and March 31 th 2019 at Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine were enrolled.The chemosensitivity of commonly used antitumor drugs was investigated and analyzed by detecting the target gene or protein with immunohistochemistry, real time fluorescence quantitative polymerase chain reaction and next generation sequencing.The tumor response rates and survival rates of patients with refractory or relapsed malignant solid tumors in individualized therapy group were compared with that in palliative therapy group. Results:The chemosensitivity of commonly used antitumor drugs was investigated in 172 patients.When other tissues were collected after neoadjuvant chemotherapy in 159 cases, the tumor tissues were collected before primary chemotherapy in 13 cases.The results of chemosensitivity in whole patients suggested the natural resistance of tumor cells to commonly used antitumor drugs except Methotrexate.The resis-tance rate of anthracyclines increased from 41.9% to 78.3% after chemotherapy ( P<0.05). Both the median survival time [(6.0±4.6) months] and the expected 2-year survival rate [(39.2±22.6)%] of individualized therapy group were higher than the median survival time [(1.5±1.4) months] and the expected 2-year survival rate (0) of palliative therapy group in 29 children developing primary and refractory disease (all P<0.05). The median survival time [(6.0±2.3) months] of the individualized therapy group was higher than that [(1.0±0.5) months] of palliative therapy group in 29 children suffering from relapsed disease ( P<0.05). However, there was no significant diffe-rence of expected 2-year survival rate between the 2 groups ( P=0.292). Conclusions:An attempt may be made to expand the application scope of Methotrexate in therapy of children suffering from malignant solid tumors.Individualized therapy based on the target detection of chemosensitivity can increase the overall survival rate of patients with refractory malignant solid tumors.However, treatment methods for children with recurrence disease still need further researches.

[Objective]To investigate the cytotoxicity of cobalt(Co2+) and chromium(Cr3+)ions on mice osteoblast(MC3T3E1)and the effects on the secretion of RANKL/OPG from osteoblast because of the stimulation of Co2+ and Cr3+ ions. [Methods]The osteoblast in vitri was cultured.The cell viability was assured by MTT test.ELISA method was applied to detect RANKL(receptor activator of nuclear factor-kB ligand),OPG(osteoprotegerin) content in serum supernatant.[Results]Compared to the control,MTT test demonstrated that Co2+ and Cr3+ ions can obviously decrease the cell viability of osteoblast.When osteoblast were exposed to Co2+ and Cr3+ ions for 24 and 48 hours,the releasion of OPG increased by 32.1% and 17.8% as compared with the control(P

OBJECTIVE: To establish the stable and efficient hearing damage model by using low dosage of cispla tin, and investigate the mechanism. METHOD: C57 mice were divided into 7 groups (every group, n = 8), the first group was the control group, the others were separately intraperitoneally injected with different dosages of cispla tin for different time. We measured the auditory brainstem response (ABR) of the mice, and obtained the basal coil of organ Corti. We observed the shape of inner hair cells (IHC) by staining AgNO3 and marked otoferlin in the IHC by immunofluorescence,successively sliced by laser confocal microscopy. The RNA fragments were amplified by RT-PCR. RESULT: After cisplatin administration for four days, the thresholds of the ABR improved in 1.5 mg/kg and 3.0 mg/kg group, and compared with the control group, the ABR thresholds improved in each group with ciplatin administration for seven days. With the same dosage, the ABR threshold of the 0.75 mg/kg x 7 d group was higher than 0.75 mg/kg x 4 d group, and there was no time-effect relationship existing in other groups with different dosage. The otoferlin was less expressed 3.0 mg/kg groups than the control group. However, the oto ferlin expressed in the 1.5 mg/kg groups were almost the same as the control group. The alteration of the IHC was observed most remarkablely in 3.0 mg/kg x 7 d group. CONCLUSION Low dosage of cisplatin can impair the hearing, and the expression of otoferlin may involve in the underlying mechanism.
Animals ; Cisplatin ; toxicity ; Cochlea ; drug effects ; metabolism ; pathology ; Hair Cells, Auditory, Inner ; drug effects ; pathology ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred C57BL

Objective:To investigate the effect of miRNA-5089-5p (miR-5089-5p) on proliferation and migration ability of esophageal cancer in vitro and its relationship with the expression of cathepsin B (CTSB) gene.Methods:Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-5089-5p in 31 tissue samples from patients who underwent esophageal cancer resection and the corresponding pericarcinomatous tissues between in March 2017 and in December 2019 at Huangshi Central Hospital of Edong Healthcare Group, and TE-13, EC9706, Eca109, KYSE30 cell lines and normal esophageal mucosal epithelial HET-1A cells. The esophageal cancer cells with the lowest expression level of miR-5089-5p were divided into 2 groups: miR-5089-5p group transfected with miR-5089-5p mimics and the negative control group with negative control sequence. qRT-PCR was used to detect the expression level of miR-5089-5p after transfection for 48 h. CCK-8 method and scratch healing test were used to detect the proliferation and migration ability of cells in the two groups. The online tools microRNA.org and Deepbase v2.0 were used to predict the target genes of miR-5089-5p. The dual luciferase reporter gene assay was used to verify the target gene of miR-5089-5p. qRT-PCR and Western blot were used to detect the expression level of target genes in the two groups. The expressions of cell proliferation-related protein (PCNA and Ki-67) and migration-related protein (N-Cadherin and Twist) were detected by using Western blot.Results:The relative expression level of miR-5089-5p in esophageal cancer and pericarcinomatous tissues was 1.54±0.53 and 7.07±1.25, respectively ( t = 24.06, P < 0.01). The relative expression level of miR-5089-5p in the esophageal cancer cell lines was lower than that of normal esophageal mucosal epithelial HET-1A cells (all P < 0.05), and the cell line with the lowest relative expression was Eca109 cells (0.12±0.03). Compared with the negative control group, the proliferation ability of Eca109 cells in miR-5089-5p group was gradually reduced with the transfection time extension, and the difference was statistically significant between the two groups since 48 h (all P < 0.05), and the migration ability was also reduced [scratch healing rate: (29±5)% vs.(64±8)%, t=3.91, P < 0.01]. The online tool predicted that the target gene of miR-5089-5p might be CTSB, and the dual luciferase reporter gene assay confirmed that miR-5089-5p complemented CTSB 3'UTR. qRT-PCR results showed that compared with the negative control group, the relative expression level of CTSB mRNA in Eca109 cells of miR-5089-5p group was reduced (0.23±0.04 vs.1.01±0.09, t = 8.27, P < 0.01). Western blot results showed that the expression level of CTSB protein was reduced, and the expression levels of cell proliferation-related protein PCNA, Ki-67 and cell migration-related protein N-Cadherin, Twist were also reduced. Conclusions:The expression level of miR-5089-5p in esophageal cancer tissues and cell lines is low. miR-5089-5p can inhibit proliferation and migration of esophageal cancer Eca109 cells. The mechanism may be achieved by down-regulating CTSB gene expression.

It has been reported that celastrus orbiculatus has anti-cancer activities. The mechanism mainly lies in its functions of inhibiting tumor cell proliferation, inducing tumor cell apoptosis, repressing tumor angiogenesis, and reversing tumor multidrug resistance, etc. To study the material basis of its anti-cancer pharmacodynamics and the mechanism of tumor inhibition has significant meanings and a wide application perspective.

Objective To take Ebola virus disease (EVD) as an example and analyze the capability of responding to emerging infectious diseases among medical university students in Chongqing city.Methods Medical university students of 2 094 in Chongqing were selected and investigated by cluster sampling.Questionnaire content includes social demographic information,knowledge,attitude,practice (KAP) and related factors about EVD.The variables of the respondents were described using mean and standard deviation for continuous variables and frequency distributions for categorical variables.Influencing factors were analyzed by using multiple-linear regression model.Results The average scores of KAP regarding EVD were 9.51 ±-3.97,5.93± 1.85,3.35 ±1.43,respectively.Multiple-linear regression analysis showed that the influencing factors of KAP regarding EVD were gender (x2 =0.773,P=0.000),residence (x2 =0.886,P=0.014) and health habits (x2 =0.316,P=0.008);gender (x2 =0.474,P=0.000),grade (x2 =0.118,P=0.024),residence (x2 =0.401,P=0.016) and health habits (x2 =0.307,P=0.000);gender (x2 =0.223,P=0.001),major (x2 =0.152,P=0.000) and health habits (x2 =0.231,P=0.000).Conclusion The capability of responding to emerging infectious diseases is not optimistic among medical university students.Medical universities should perform effective intervention according to the characters of different clusters.

Objective To determine the plasma concentration of cytarabine(Ara-C) in children with leukemia and obtain dynamics parameters, and investigate the relationship between the parameters and clinical effect in order to provide the basis for optimization of Ara-C application. Methods Using highperformance liquid chromatogram (HPLC) to determine the plasma concentration of Ara-C, its metabolite Ara-U and infusion rate in 37 children with acute leukemia, their therapeutic reaction, remission, treatment-related infection, side-effect and long-term treatment effect were analyzed in statistic. Results Ara-C by 1～2 g/m2 intravenous drop infusion for 2 hours, the peak plasma concentration time was 2 h and peak concentration were (14.37-84.44)μmol/L, and the median was (41.42±22.80)μmol/L. The median infusion rate was 869.57at 30 minutes after Ara-C drip completion, its average level was (253.40±81.49) μmol/L, over six-times than Ara-C peak concentration. The median continuous complete remission time in 37 children was 29.8 months (5.0～53.1 months), 3y-DFS was (90.63±5.15)%. The therapy-related infection rate was 56.8 %(21/37),including three children (8.1 %) suffered from severe infection, but there was no therapy-related death and no children were off the protocol due to poor tolerance. Conclusion As post-remission treatment, high-dose Ara-C would not cause cumulation in vivo in children with acute leukemia and side-effect were slight. Ara-C could improve the long-term continuous complete remission rate and clinical cure rate for children with leukemia. Therefore, it was worth to apply in clinical.

Objectives To investigate the clinical profile and prognosis of hemophagocytic syndrome (HPS). Methods A retrospective study was carried out to analyze the clinical features and laboratory findings in 28 children with HPS. Fisher's exact probability method and Logistic multivariate regression were used to explore the prognostic risk factors.. Results HPS was clinically characterized by prolonged fever (100%), hepatomegaly (64.29%),and other minor features including respiratory symptoms (53.57%), splenomegaly (50%), hydrops of multiple serous cavity (42.86%), lymphadenectasis (32.14%), jaundice (17.85%), skin rash (14.29%), central nervous system involvement (14.29%), and alimentary tract hemorrhage (10.71%). Labo-ratory data showed that 1iver dysfunction, pancytopenia, coagulation abnormalities, disseminated intravascular coagulation, hy-pertriglyceridemia, decreased number of natural killer cells and hyponatremia were prominent. The etiological analysis indicated that infection associated hemophagocytic syndrome was most common (60.71%), in which EB virus associated HPS was pre-dominant, accounting for 64.71%. Significant difference was observed in the difference of albumin,blood urea nitrogen and acti-vated partial thromboplastin time between death and survival cases (P<0.05). The Logistic regression multivariate analysis showed that hypoalbuminemia was an independent prognostic factor. Conclusions There are various underlying diseases and clinical manifestations for HPS. The lower level of serum albumin is an independent prognostic factor. A prompt diagnosis and treatment is very important for HPS prognosis due to the rapid progression and high mortality.

Objective To investigate the effects of all trans-retinoic acid (ATRA) on the expressions of iron metabolism-related genes and their products in K562 cells and the possible relationship. Methods (1) The characteristics of K562 leukemic cell differentiation induced by ATRA was evaluated by Benzidine, Wright's, NSE and NBT staining.(2) The expression levels of cellular surface antigens (CD71 and CD 13) in K562 cells cultured with ATRA were measured by flow cytometry. (3) IRP/IRE binding activity was assessed by RNA/protein band-shift assay.(4) Ferritin was determined by radioimmunoassay.(5) The mRNA expression levels of H-Fn, TfR and IRP2 in K562 cells cultured with different concentrations of ATRA were delineated by RT-PCR method, confirmed by sequencing of RT-PCR products. Results K562 cells could be induced to differentiate into neutrophils by ATRA, confirmed by cytochemical staining. The expression of CD71 decreased while CD13 increased. The mRNA expression levels of TfR and IRP2 were decreased while mRNA expression level of H-Fn was increased in K562 cells cultured with ATRA, compared to that in control cells. Concomitantly,IRP binding activity was significantly decreased but the level of ferritin was significantly increased in K562 cells cultured with ATRA. Conclusions During the course of K562 cells induction and differentiation to myelocytes by ATRA, the expression level of iron metabolism-related genes and products were changed but the upstream-regulation mechanism still remains unclear.