Background and purpose:Apollon gene is highly expressed in leukemia and other tumors. The study aimed to discuss whether RNAi technology can reverse multidrug resistance of chronic myeloid leukemia cell line K562 through constructing a eukaryotic vector of short hairpin RNA (shRNA) targeting at Apollon gene. Methods:The eukaryotic vector pGPHI-GFP-Neo-Apollon with shRNA targeting at Apollon gene was constructed and then transfected into K562 cells by LipofectamineTM2000, and G418 pressure selection. Reverse transcription-polymerase chain reaction (RT-PCR) and immunolfuorescence were used to detect the expression of Apollon mRNA and protein after Apollon was transfected stably in K562 cells. The changes of sensitivity of K562 cells to leurocristine (VCR) and etoposide (VP16) after transfection with shRNA-Apollon were detected by MTT method, and the apoptosis rate was detected by flow cytometry. Results: pGPHI-GFP-Neo-Apollon carrier was constructed successfully and expressed stably in K562 cells, and after G418 screening, it silenced Apollon mRNA and protein expression effectively. According to the result of MTT, the sensitivity of K562 cells to VCR and VP16 increased significantly in the group of gene interference, with half of its inhibition concentration (half-inhibitory, IC50) value signiifcantly lower than the control group (P<0.05);Flow cytometry showed that the cell apoptosis rate was increased signiifcantly (P<0.05), but there was no statistically signiifcant difference in the apoptosis rate between shRNA negative control group and normal control group (P>0.05). Conclusion:pGPHI-GFP-Neo-Apollon carrier can enhance the abilities of VCR and VP16 to induce the apoptosis of K562 cells, namely an increase of sensitivity to these chemotherapeutics in K562 cells, it is hinted that RNA interference targeting Apollon gene may reverse the multidrug resistance of leukemia cells in some degree.

[Summary] The paper reported 9 cases of external type cesarean scar pregnancy treated with transvaginal local lesion resection between January 2013 and March 2014 in this hospital .The surgical exposure was established by using vaginal tractors .The cervical front lip was pulled below outwards by using cervical clamp to expose the front vaginal fornix .Hemostatic water was injected into the cervix vaginal clearance (epinephrine 0.5 mg +NS 500 ml).The vaginal mucosa was opened at bladder cervix groove level for entering the bladder cervix clearance .The bladder was seperated upwards and to the side with fingers to expose uterine isthmus lesions . After cutting the thin muscle , the embryos and part of necrotic tissues were extracted for pathological examination .The incision scar around tissues were trimed and a drainage tube was placed in the uterine .A myometrium full-thickness continuous suture was performed with 1-0 absorbable sutures ( paying attention to avoid endometrial tissues ) , and the vaginal wall was continuously sutured with 1-0 absorbable thread .The drainage tube was removed 5 days after operation .The operations were successfully accomplished in all the 9 cases.The average operation time was 65 min (40-90 min), and the average blood lose was 60 ml (40-80 ml).Cervical tube adhesion occurred in 1 case, and no complications were observed in the remaining cases .The average hospitalization time was 7.5 days (5-10 days).The postoperative time of menstruous restoration was 28-40 days.We deem that transvaginal resection of cesarean scar pregnancy lesion is simple and deserves clinical popularization .

Objective To discuss the intervention effect of four oral care model on compliance of atomizing inhalation and mouth and throat complications in outpatient infants and young children patients. Methods A total of 480 cases of children patients using budesonide inhalation for the treatment of respiratory tract infection were divided into the observation group(n=260)and the control group (n=220) according to the time sequence. The control group received conventional nursing, the observation group used quaternity oral care model in nursing care. The compliance and occurrence of complications in the mouth and throat were observed between the two groups. Results In the observation group with nebulae inhalation compliance was 79.2% (206/260), evidently higher than that in the control group the compliance of 58.6%(129/220) (χ2=23.97,P<0.01). The incidence of oral herpes and ulcers in the control group was 2.73%(6/220), and 0.77%(2/260) in the observation group, but no significant difference was seen (χ2=2.79, P>0.05). The incidence of thrushes was 3.64%(8/220) in the control group, and none occurred in the observation group, which showed significant difference (χ2=9.62, P<0.01). Conclusions Using quaternity oral care model in corticosteroid nebulae inhalation patientscan increase compliance and decreasecomplications of oral and throat of outpatient children.

Objective To study the dose-dependent effect of sodium arsenite on the expression of p53 gene in human embryonic lung fibroblasts(HELF)in vitro.Methods The human embryonic lung fibroblasts were divided into four groups in vitro based on completely randomized design.The expression of p53 mRNA was detected by real-time PCR and the expression of p53 protein was detected by immunohistochemical SAB,respectively,in human embryonic lung fibroblasts which were exposed to different doses(0,3,9 and 15?mol/L)of sodium arsenite for 24 hours.The one-way analysis of variance and post hoc comparisons were performed for testing the differences among groups and the linear correlation was for testing correlation between doses and the expression of p53 gene.Results Compared with that of exposure to 0?mol/L sodium arsenite,the expression of p53 mRNA in HELF was increased significantly(P0.05).The p53 mRNA expression was increased in dose-dependent manner with the increased concentration of sodium arsenite(r=0.947,P

Objective To study the pathology of primary neuroectodermal tumor (PNET),and its diagnostic standards and prognostic factors.Methods Expression of CD99,FLI-1,Syn,NSE,S-100,NF,and Vim was detected in PNET tissues stained with HE and immmunohistochemistry (En Vision method).Survival rate of 33 PNET patients with complete clinical information was analyzed with COX regression analysis method.Results The positive expression rate of CD99,FLI-1,Vim,Syn,NSE and s-100 was 88.57%,51.43%,91.42%,48.57%,45.71%,and 22.86%,respectively.The sensitivity of combined CD99 and FLI-1 was 100%.However,NF was not expressed in all PNET tissues.When other factors were unchanged,no difference was found in the effect of age on the survival rate of patients.However,a significant difference was observed in the effect of PNET site and its treatment modalities on the survival rate of patients (P

Objective To screen siRNAs that can effectively inhibit Apollon gene expression and determine the cellular functions of those siRNAs.Methods A chemical synthesis method was used to synthesize 3 siRNA sequences against different sites of Apollon.They were transfected into the human breast cancer MCF-7 cells by using Lipofectamine 2000.mRNA level of Apollon was determined by reverse transcription-polymerase chain reaction (RT-PCR).Cellular immunity fluorescence quantitative analysis combined with confocal laser technology was used to determine the protein level of Apollon.Methyl thiazolyl tetrazolium bromide (MTT) assay and flow cytometry were used to determine the effects of siRNA targeting Apollon on proliferation and apoptosis of MCF-7 cells,respectively.Results Three pairs of siRNA could significantly inhibit Apollon mRNA expression,at the inhibition rates of (36.201±11.629) ％,(67.308±7.686) ％and (47.123±12.000) ％,respectively (P ＜ 0.05).After tranfection by siRNA2,Apollon protein fluorescence intensity was (14.97±2.08) ％ compared with control cells.The cell proliferation MCF-7 was inhibited by (73.361±2.118) ％and apoptosis was increased by (28.793±0.743) ％.Conclusions Screened siRNA2 effectively silences Apollon gene expression,effectively inhibits the proliferation and increases the apoptosis of MCF-7 cells.This provids the foundation for its clinical application in cancer therapy.

The purpose of this study is to prepare galactosyl modified lipid bilayer-coated mesoporous silica nanoparticles (GPEM) to enhance the antitumor efficacy against hepatocellular carcinoma cells. The irinotecan (CPT-11) loaded mesoporous silica nanoparticles (MSNs) was coated with the Gal-P123 modified functional lipid bilayer by thin-film dispersion method. Nanoparticles were characterized with particle size, zeta potential, morphology and drug release in vitro. Afterwards, the cell uptake, intracellular concentration of CPT-11, cell apoptosis rate and cytotoxicity were evaluated on human hepatocellular carcinoma cell line Huh-7. The results showed that MSNs were coated with intact lipid bilayers and the nanoparticles had clear core-shell structure. GPEM is stable with the mean particle size of (78.01 +/- 2.04) nm. The low leakage rate in normal physiological conditions in vitro is contributed to the protection of stable lipid bilayer, and the fast drug release in acid environment due to the destruction of the lipid bilayer. On the cell level, the vector could improve the intracellular CPT-11 concentration by 4 times because of the functional lipid bilayer. The high CPT-11 concentration led to the increasement of apoptosis rate by 48.6%, and the reduction of half maximal inhibitory concentration (IC50) values of CPT-11 by 2 times, indicating stronger cell cytotoxicity.

Background and purpose:Extranodal natural killer/T-cell lymphoma (ENKTL) is a form of non-Hodgkin’s lymphoma. The ENKTL incidence in China is much higher than that in the Western countries. The disease is highly malignant, not sensitive to chemotherapy, has short survival period and poor prognosis. Epstein-Barr virus (EBV) infection has close relationship with the development of the disease. However, there are still a few patients without EBV infection. This study aimed to discuss the clinical features and prognosis of EBV-encoded small RNA (EBER) in situ hybridization negative ENKTL.Methods:From Aug. 2011 to Oct. 2015, 326 cases were diagnosed with ENKTL from the First Affliated Hospital of Zhengzhou University. The expression of EBER was detected by in situ hy-bridization technique. The clinical pathological characteristics and prognosis of EBER-negative patients were analyzed. Results:In 326 patients with ENKTL, the negative rate of EBER was 2.45% (8/326). In 8 EBER-negative patients, the median survival time was 17 months. The log-rank test revealed that there was a signiifcant difference between EBER-negative and EBER-positive curves (χ2=6.407,P=0.011). Multivariate Cox proportional hazards regression analysis showed that in EBER-negative ENKTL, only lactate dehydrogenase (LDH) predicted survival time (P=0.008). EBV-DNA copy number in plasma was not signiifcantly correlated with survival time (P>0.05).Conclusion:The inci-dence of EBER-negative ENKTL is low. Patients with EBER-negative ENKTL have poorer prognosis than EBER-posi-tive patients. Elevated LDH may be a factor indicating poor prognosis.

Objective:To analyze clinical characteristics, treatment, and prognosis of B-cell lymphoma, unclassifiable, with features in-termediate between diffuse large B-cell lymphoma and Burkitt lymphoma (DLBCL/BL). Methods:The clinical and pathological data of 13 DLBCL/BL patients, who were treated in the First Affiliated Hospital of Zhengzhou University between January 2013 and December 2014, were collected. Overall survival (OS) and progression-free survival (PFS) were estimated by the Kaplan-Meier method. Through the log-rank test, survival curves were compared among groups classified by clinical stage, age, serum lactate dehydrogenase (LDH) lev-el, international prognostic index (IPI) score, or first chemotherapy regimen. Results:Among the 13 patients with DLBCL/BL, 12 pa-tients showed extra-nodal involvement. The median OS and PFS were only 10 and 6 months, respectively. Univariate analysis showed that the LDH levels and IPI scores exerted statistically significant effects on prognosis. Some borderline differences in survival were not-ed among the CHOP, CHOP-like, and intensive chemotherapy groups. Conclusion:DLBCL/BL is an aggressive B-cell lymphoma with a short survival time. The majority of patients presented extra-nodal involvement. DLBCL/BL did not respond well to CHOP or CHOP-like regimen, and more intensive chemotherapy may improve survival. Elevated LDH levels and high IPI scores were predictors of poor sur-vival.

Objective To explore the clinical characteristics of the lesions of spinal cord and optic nerve as the onset of neuromyelitis optica (NMO).Methods A total of fifty-one patients with myelitis or optic neuritis (ON) as the onset of NMO who hospitalized in our Neurology Department during October 2010 to October 2012 were enrolled in the study.Clinical presentations and MRI findings of the spinal cord and brain were studied.Results (1) A total of 26 cases (51.0％) presented with myelitis as the index event,in which 30.8％ (8/26) were the longitudinally extensive transverse myelitis (LETM) and 69.2％ (18/ 26) were non-LETM (short segmental myelitis or non-transverse myelitis).Patients with non-LETM as the onset were found to have better prognosis than those with LETM (full recovery ratio was 13/18 vs 2/8,P ＜ 0.05),while shorter recurrence interval of myelitis and higher recurrence frequency of events were shown in patients with non-LETM (11.1 vs 18.6 months,3 times per year vs once per year,with all P ＜ 0.05).(2) A total of 25 cases (49.0％) presented with ON as the index event with 24.0％ (6/25) of unilateral ON and 76.0％ (19/25) of bilateral ON.Patients with bilateral ON had more severe visual impairment and shorter first remission period than those with unilateral ON (P ＜ 0.05).Conclusion Non-LETM and bilateral ON are the most common index demyelinating events in NMO cases.