Objective To observe the changes of platelet activation markers in newborn infants with hyperbilirubinemia,and its relationship with myocardial enzymes and clinical significance.Methods Thirty neonates with hyperbilirubinemia were selected as observation group,and 15 health newborns were served as normal control group.In the morning,1 mL fasting blood in the femoral vein was collected from the patients in both groups,and 20 g/L EDTA-Na2 0.1 mL anticoagulant was added with a gentle shake.CD41-FITC 10 ?L was injected into two test tubes,with IgG1-PE and CD62-PE reagent 10 ?L added,diluted liquid 200 ?L PBS,and with 5 ?L whole blood,under dark room temperature for 15 minutes.Negative control tubes(CD41-FITC plus IgG1-PE) were applied to adjust voltage,the flow cytometry was used to determine CD62-PE.Myocardial enzymes and liver function were measured by automatic biochemistry analyzer.Results The levels of platelet activation markers in observation group were significantly higher than that in the normal control group(P

Controlled Ecological Life-support System(CELSS)is one of the key technologies that must be solved before founding permanent base in space such as Lunar or Mars base.After discussing with Russian experts and analysing the data,the author introduced something important about CELSS in Russia,including research methods,achievements and problems to be solved urgently.In the end a proposal for developing CELSS research in China was given based on them.

ObjectiveTo observe the effect of mild hypothermia on vital signs of the elderly patients with anterior circulation brain infarction (massive hemispheric infarction) and hyperpyrexia.Methods46 elderly patients with massive hemispheric infarction and hyperpyrexia within the first 24 hours after onset were randomly divided into the control group and the hypothermia group with 23 cases in each group. The patients of the hypothermia group were treated with mild hypothermia therapy (33℃～35℃). Those of the control group were treated with routine hypothermal treatment. The temperature and heart rate of all patients were measured after 10 days; the prognosis was evaluated according to mortality and scores of Neurological Deficit Scale (NDS).ResultsFor patients of the hypothermia group, the temperature and heart rate significantly reduced, death rate was significantly lower, the neurological function distinctly improved and prognosis was better than the control group after 10 days treatment ( P<0.05).Conclusion Mild hypothermia is safe and effective to elderly patients with massive hemispheric infarction and hyperpyrexia.

Objective To detect the microsatellite instability (MSI) of D9S166,D9S171,D9S941, D9S942 and IFNA located at chromosome 9p21 in gastric carcinoma with high-grade intraepithelial neoplasia and normal tissue,and investigate the correlation of p14,p15,p16 gene and the gastric carcinogenesis. Methods 55 cases of gastric carcinoma with high-grade intraepithelial neoplasia and normal tissue were se- lected to detect MSI by microdissection,polymerase chain reaction,denaturing polyaerylamide gel elec- trophoresis and silver nitrate staining technology of five microsatellite markers on 9p21.Results In the infor- mative cases,total ratio of MSI in gastric carcinoma was 27 % (64/233) and it was 18% (42/233) in high- grade intraepithelial neoplasia.The ratio of MSI was significantly higher in gastric carcinoma than that in high-grade intraepithelial neoplasia (P

BACKGROUND: The existence of minimal residual leukemia cells is the main cause for the recurrence of acute leukemia in children, and immunological biological therapy has attracted more and more attentions in the various methods from eliminating minimal residual disease. Previous studies have found that interferon α-2b can effectively inhibit the increase of tumor cells in vivo in children with neuroblastoma and malignant lymphoma, whether it can inhibit the increase of leukemia cells?OBJECTIVE: To investigate the effects of interferon α-2b in vitro on leukemia cells.DESIGN: A comparative observation taking human promyelocytic leukemia HL-60 cell line as the material.SETTING: Cell Culture Room; Immunological Laboratory; Cell Room, Institute of Pediatrics, Affiliated Hospital,Medical College of Qingdao University.MATERIALS: HL-60 cell line was provided by Shandong Institute of Basic Medical Sciences. Interferon α-2b was purchased from Megagene Company Fluorescein isothiocyanate (FTTC) rabbit-anti-rat Ig solution (CatEK001) and CD13 anti-human monoclonal antibody solution (Cat. DK013Y) were purchased from Union Stem Cell & Gene Engineering Co.,Ltd.METHODS: The experiments were carried out in the Institute of Pediatrics, Affiliated Hospital, Medical College of Qingdao University from March to September 2005. HL-60 cells culture system was established in vitro, and the oncentration was adjusted to 1×109 L-1. The cells were divided into control group and experimental group. In the experimental group, each well was added by interferon-α-2b with the terminal concentration of 5×105, 1×106, 2×106,5×106 and 1×107 U/L, respectively. In the control group, each well was added by saline of the same volume. The cells were cultured continuously for 48 hours. The morphological changes of HL-60 cells were observed using Wright's staining under light microscope; Cell apoptosis was observed using acridine orange/ethidium bromide double staining; Antigen expression and maturation and differentiation on cell membrane were observed by determining CD13 protein expression; Proliferation and activity of HL-60 were detected with methyl-thiazol-tetrazolium (MTT) assay.MAIN OUTCOME MEASURES: The occurrence of apoptosis was judged according to the uniformity and staining of HL-60 nuclear chromatin; HL-60 cell proliferation was judged according to the absorbance (A) value; The maturation of HL-60 cells was judged according to the number of positive CD13 cells.RESULTS: ① HL-60 cell apoptosis: The cells were cultured for 48 hours. When the concentration of interferon α-2b was 5×105 U/L, there were mainly early apoptotic HL-60 cells; When the concentration was 1×107 U/L, there were mainly late apoptotic cells, and the apoptotic rate was significantly higher than those in the control group (P ＜ 0.01 ).② HL-60 cell proliferation: The A values in the experimental groups treated with interferon α-2b of 2×106 U/L and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01). ③ Maturation of HL-60 cells: The percentages of positive CD13 cells in the experimental groups treated with interferon α-2b of 1 ×106 and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01).CONCLUSION: It is concluded that interferon α-2b can enhance the apoptosis, inhibit the proliferation and promote maturation and differentiation of HL-60 cells.

Objectives:To explore the prevalence and correlates of attempted suicide among Beijing psycho-logical aids hotline callers,in order to provide knowledge of hotline-based suicide prevention.Methods:From De-cember 2002 to December 2008,all the effective callers who consulted themselves'psychological problem,by Bei-jing psychological aids hotline were interviewed by professional hotline operators during the consulting calling.Data of whether callers attempted suicidal behavior in 2 weeks prior to the calling,gender and other 5 demographic char-acteristics of callers were collected via hotline interview.A questionnaire including depression screening scale,com-mon psychiatric symptoms such as delusion,hallucination,etc.,a history of substance abuse or dependence,a histo-ry of suicidal behavior,having had acute or chronic life events,and severity of hopeless was used during hotline in-terview.Logistic regression model was used.Results:In 22 415 interviewed callers,362 (1.6%)callers (233 fe-males)reported that they attempted suicidal behavior in 2 weeks prior to the calling.After adjusted for demographic characteristics,prior suicidal behavior 2 weeks before calling (OR=4.43,95%CI:3.37-5.83),hopelessness (OR=2.01,95%CI:1.52-2.67),substance abuse (OR=1.97,95%CI:1.48 -2.62),acute negative life events (OR=1.94,95%CI:1.49-2.52),and severe depressive symptoms (OR=1.45,95%CI:1.08-1.95)were associatedwith attempted suicide among hotline callers.Conclusion:The rates of attempted suicide among hotline callers are probably high.Suicide prevention in hotline would shed light on improving coping skills for life events and sub-stance abuse problems for hotline callers.

Objective:To develop the Counseling Skills Rating Scale for Psychological Aids Hotline (CSRSPAH),an instrument to objectively assess the quality of process of counseling or intervention provided by hotline operators,and to test its validities and reliabilities.Methods:A draft of the scale,which consisted of 50 items under 5 dimensions,was designed in line with previous researches and the practice.In pilot study,supervisors at Beijing psychological aids hotline,assessed tape recorded sessions and gave feedbacks on the draft of the scale.The scale had been revised based on the pilot study.Twenty-eight items were deleted,and several items were rephrased.Finally,a scale which consisted of 22 items within 3 dimensions,i.e.counseling process,attitude,and communication skill,was developed.The scoring standards of the scale were also developed.To evaluate the IntraClass Coefficients (ICC) of the CSRSPAH,each of the tape records of 37 callings from 2005 to 2007 were assessed by 7 supervisors independently,using the scale.And the tape records of other 318 callings from 2013 to 2014 were also assessed by supervisors,using the CSRSPAH.The results of the 318 assessed callings were used to test the construct validities with the Confirmative Factor Analysis.The Cronbach a coefficients of the total score and three dimensions,the discriminant indices of every items,and correlations of each items and each dimensions were calculated,based on the 318 assessed callings.Results:One of the items (referral) was deleted due to excessive amount of missing data.Results of cortfirmative factor analysis of the remained 21-item scale revealed that the 3-factor construct structure of the scale was robust.The fitting indices of the confirm factor analysis were,x2/df=675.21/186,CFI =0.92,NNFI =0.91,RMSEA =0.10,SRMR =0.08.The Cronbach α coefficients of the total score,scores of counseling process,attitude,and communication skill,were 0.89,0.68,0.81 and 0.77,respectively.The ICCs of the inter-rater reliabilities of the total score and 3 dimensions of the scale were 0.67,0.59,0.59,and 0.67,respectively.The discriminant indices of all the 21 items ranged from 0.09 to 0.32.The correlation coefficients of scores of each items and scores of 3 dimensions and total scores were greater than 0.30,and reached statistical significance.Conclusion:The validities and reliabilities of the Counseling Skills Rating Scale for Psychological Aids Hotline are acceptable.The scale could be used in assessing the quality of hotline counseling or intervention,and related studies in the future.

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

OBJECTIVETo investigate whole blood viscosity changes at different time points in rats model with lumbar vertebrae semidislocation, study Shi's theroy of qi and blood and "Gucuofeng and Jinchucao" [symbols: see text], also reveal pathological physiology characteristics of spinal disorder.

METHODSThirty-six SPF male rats weighted 350 to 450 g were randomly divided into rotatory fixation group (RF group), simple fixation group (SF group) and Sham group (Sham group), 12 rats in each group. Exterior vertebrae implanted through L4-L6 segments of lumbar vertebrae in RF and SF group were connected fixed device. In RF group, L5 spinous process were rotated to right, and caused L5 spinous process was non collinear with L4 and L6; in SF group, external fixed device were simple connected without rotation. At 1, 4, 8 and 12 weeks after fixation, whole blood viscosity changes were tested.

RESULTSAt 4 and 8 weeks after fixation, high (150/s), medium (60/s) and lower (10/s) shear rate in RF and SF group were higher than that of Sham group (P<0.05). At 1 and 12 weeks, there was no sigificant differences among three groups in whole blood viscosity (P>0.05).

CONCLUSION"Gucuofeng and Jinchucao" [symbols: see text] vertebrae could raise whole blood viscosity, increase degree of bloos stasis and induce or aggravate spinal disorder in further.

Animals ; Blood Viscosity ; Disease Models, Animal ; Joint Dislocations ; surgery ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVETo construct a lentiviral vector of repressor element-1/neuron-restrictive silencer element (RE-1/NRSE) double-stranded RNA (dsRNA).

METHODSThe RE-1/NRSE cDNA containing both sense and antisense oligo DNA fragments of the targeting sequence was synthesized and cloned into the pGC-LV vector. The obtained lentiviral vector containing RE-1/NRSE dsRNA was confirmed by PCR and sequencing. A total of 293T cells were cotransfected with lentiviral vector of L-smNRSE/RE-1, pHelper 1.0, and pHelper 2.0. The titer of virus was measured based on the expression level of green fluorescent protein. The transfection efficiency of green fluorescent protein into rat mesenchymal stem cells was calculated.

RESULTSPCR and DNA sequencing demonstrated that the constructed lentivirus vector of L-smNRSE/RE-1 produced RE-1/NRSE dsRNA.The titer of the concentrated virus was 4x108 TU/m1. The virus was stably transfected into rat mesenchymal stem cells, and the infection efficiency reached 100% when the multiplicity of infection was 80.

CONCLUSIONThe lentivirus vector of RE-1/NRSE dsRNA is successfully constructed.

Animals ; Bone Marrow Cells ; Cells, Cultured ; Genetic Vectors ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; Plasmids ; genetics ; RNA, Double-Stranded ; genetics ; Rats ; Repressor Proteins ; genetics ; Silencer Elements, Transcriptional ; genetics ; Transfection