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.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Aged ; Catgut ; Female ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; physiopathology ; Spasm ; physiopathology ; therapy ; Treatment Outcome

Objective To study the ultrasonographic characteristics of Wilms′tumor (WT), aims at elevating the diagnostic level.Methods We retrospectively studied sonographic features in 30 cases of WT proved by operation and pathology.Results The tumors with heterogeneous pattern of solid and cystic tissue were shown in 22 cases.Seven cases demonstrate predominantly echogenic tumor. A huge cystic mass with numerous septas was shown in 1 case. The tumor extended into renal pelvis in 4, into renal vine in 1.The tumor metastasized to lymphnod in 1, to bilateral lungs in 1. Among 30 cases, the tumor was huge and grew beyond the renal profile in 26 cases, in 3 cases the tumors took up the renal sinus and made its structure turbulence, the renal shape was approximately normal.One case was extrarenal WT,the tumor located in retroperitonemu.Conclusions Sonographically, typical WT is a large heterogeneous mass within or without cystic areas, it also can be a pure cyst. It is easy to diagnose. When the tumor is limited to renal outline, it is easy to confusion with renal cancer. In addition, there are bilateral WT and extrarenal WT. The sonographic characteristics is similar among WT, clear cell sarcoma of kidney, malignant rhabdoid tumor and congenital mesoblastic nephroma. It is difficult to distinguish them from ultrasonography.

Objective In order to get reference data for diagnosis of clinical genetic disease through analyzing chromosome ab‐normality types and rates in 885 patients who ask for cytogenetics consultation in recent years .Methods 324 newborns who asked for cytogenetics consultation because of high risk factors in down′s screening during pregnancy or found abnormality in physical ex‐amination after birth and 561 patients with history of spontaneous abortion ,infertility or fetal death ,growth or mental retardation , sexual abnormality were examined for karyotype analysis .Results 116 cases of chromosome abnormal karyotypes were detected , count for 13 .11% ,among which ,40 cases(34 .48% ) are chromosomal aberration .Chromosome abnormality types and rates are dif‐ferent in patients with different type of diseases .Conclusion Abnormalities in chromosome numbers and sex chromosome abnor‐mality account for the main causes of growth and mental retardation ,abnormalities of sexual differentiation and development ;but for couples suffered from spontaneous abortion ,polymorphism and structure abnormalities takes up the highest portion .

Objective To explore the value of multivoxel 1H-MRS in the differential diagnosis of corpus callosum lesions.Methods 17 patients with corpus callosum lesions confirmed by clinical or pathological method underwent MRS examination.These patients included 2 cases of lymphoma,4 cases of Marchiafava-Bignami disease,3 cases of glioma, 7 cases of infarction,and 1 case of multiple sclerosis.MRS features of lesions were analyzed.Results 2 cases with corpus callosum lymphoma revealed significantly increased Cho peak, decreased Cr and NAA peak,towering Lip peak.Among 4 cases with Marchiafava-Bignami diseases, 2 cases revealed increased Cho/Cr and decreased NAA/Cr,1 case showed normal,1 case revealed inverted Lac peak.For 3 cases with corpus callosum glioma,NAA peak decreased or disappeared,and Cho peak increased in varying degrees.7 cases with corpus callosum infarction revealed significant inverted Lac peak, and NAA peak decreased in varying degrees.1 case of multiple sclerosis revealed increased Cho peak,decreased NAA peak,and inverted Lac peak during acute period with enhancement,while these features returned normal during inactive period.Conclusion Multivoxel 1H-MRS plays an important role in the differential diagnosis of corpus callosum lesions.

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.

Background There are two types of different questionnaires in dry eye diagnosis.But the associations about two questionnaires or questionnaire and clinical examination are still unclear.To effectively quantize the symptoms is helpful for a correct diagnosis of dry eye disease.Objective This survey was to evaluate the Standard Patient Evaluation of Eye Dryness (SPEED) and Ocular Surface Disease Index(OSDI) questionnaire for the diagnosis of dry eye and investigate the correlation between the clinical examinations and questionnaires. Methods A perspective cohort study was designed.Sixty-six patients were enrolled in this study.This study was approved by the Ethic Committee of Peking University First Hospital,and written informed consent was obtained from each subject before any ocular examination.SPEED-based and OSDI-based questionnaires were used to score the dry eye symptom and grouped according to severity of complains.Corneal fluorescence staining,tear film breakup time(BUT),Schirmer I test and tear film interferometry were performed in all patients.The correlations between two questionnaires scores and their association with clinical examinations were evaluated.Results The negative correlations were found between the SPEED-based score or OSDI-based score with BUT value(r=0.390,P=0.001 ;r=-0.395,P=0.001 ),but no significant correlations were seen between the SPEED-based score or OSDI-based score with Schirmer test( r=-0.081,P=0.515; r=-0.080,P=0.525)and tear film interferometry score(r=0.158,P=0.204;r=0.219,P=0.077).The BUT was significantly prolonged in mild symptom group compared with serious group(t=2.339,P=0.022),but no significant difference was seen in Schirmer Ⅰ test and tear film interferometry scores using SPEED-based questionnaire ( t =0.404,P =0.687 ; t =- 0.947,P =0.347 ) ; while the positive fluorescence staining rate between two groups was significantly different (x2 =0.164,P =0.685 ).When using OSDI-based questionnaire,significant difference in BUT was seen among mild,moderate and serious symptom groups ( F =11.871,P =0.000 ),and BUT in mild symptom group was delayed in comparison with moderat and serious groups( P=0.000,0.000).No significant differences were found in Schirmer Ⅰ test,tear film interferometry scores and fluorescence staining rate among three groups(F=1.432,P =0.246; F =2.799,P =0.068; x2 =6.026,P =0.050).SPEED score showed a positive correlation with OSDI score ( r =0.697,P =0.000 ). Conclusions Both OSDI and SPEED are effective tools for the evaluation of symptoms of dry eye.The two types of questionnaires are consistent in symptoms evaluation.

Objective To investigate the current situation of maternal self- efficacy of breastfeeding and explore its influencing factors in Zhengzhou. Methods Using the general condition questionnaires, breast feeding self-efficacy scale, perceived social support scale and Edinburgh postnatal depression scale to assess the situation of 180 puerperas by the convenience sampling in one obstetric hospital of Zhengzhou. Results The total score of maternal breast feeding self-efficacy was(114.04 ± 21.57)points.The frequency of delivery, the way of delivery and feeding ways, the average income of family were effected with breastfeeding self-efficacy. The total score of social support was(68.87 ± 10.43) points, the total score of puerperas depression was(7.61 ± 4.25) points.The social support score had positive correlation with breastfeeding self-efficacy(r=0.423, P<0.01). Puerperas depression had negative correlation with breastfeeding self- efficacy(r=- 0.342, P<0.01). Conclusions The maternal breastfeeding level in Zhengzhou was at a lower level. The level of maternal breastfeeding self-efficacy scale was impacted by maternal social support degree, the degree of depression. Measures should be taken to enhance maternal social support degree, reduce the degree of depression, in order to improve the level of maternal breastfeeding self-efficacy.

Objective To determine the efficacy of glucose-loweringtreatment by proton magnetic resonance spectroscopy (1H-MRS).Methods Bilateral frontal lobes of 45 male patients with type 2 diabetes mellitus (T2DM) underwent 1 H-MRS separately bythe time it was first diagnosed.Among them,30 were treated with insulin,15 were rejected.The bilateral frontal lobes of all patients were re-examined with 1 H-MRS after 6 months.The ratios of metabolites in the frontal lobe were recorded and compared,and observing whether the lactate(Lac) peak appeared or not.Results Significantly higher values of the N-acetylaspartate (NAA)/creatine (Cr)ratio and NAA/choline (Cho) ratio after 6 months (2.414 2± 0.206 6 and 2.244 2±0.341 5) were found in bilateral frontal lobes compared with pre-therapy (1.905 3±0.135 7 and 1.906 0±0.322 1)(P＜0.05).Significantly lower values of the NAA/Cr ratio and NAA/Cho ratio in the refuse treatment group(1.504 2 ± 0.262 4 and 1.510 0± 0.220 8) were found in bilateral frontal lobes compared with pre therapy (1.960 0±0.175 0 and 1.880 8±0.297 0)(P＜0.05).Lac peak was not detected in the all experiments.Conclusion 1 H-MRS provides valuable information in the assessment and evaluation of clinical efficacy and prognosis of T2DM by detecting the changes of brain metabolism.

Objective To explore the effects of Sanban manipulation of spine on ankylosing spondylitis (AS) at the early-middle stage.Methods 60 AS patients at the early-middle stage were divided into drug group (n=30) and manipulation group (n=30). The drug group wastreated with sulfasalazine and the sustained-release pills of Zhengqingfengtongning, while the manipulation group were treated with Sanbanmanipulation of spine additionally. Self-developed questionnaire were used to follow up before and 1, 3 and 6 months after treatment. ResultsThe Schober index and the finger-floor distance improved more in the manipulation group than in the drug group (P<0.05) 3 months aftertreatment. All the index improved more in the manipulation group than in the drug group (P<0.05) 6 months after treatment. ConclusionSanban manipulation of spine can improve the thoracolumbar activities in the AS patients at the early-middle stage.