Using an anti-nucleolar protein B23 monoclonal antiboby, we examined the changes of protein B23 content in NIH3T3 fibroblasts during serum-induced proliferation and the difference in protein B23 content between mitotic and interphase NIH3T3 fibroblasts by immunofluorescence and im-munoblot methods. When NIH3T3 cells grew confluent, the serum containing medium was removed and replaced by serum-free medium,and the cells were cultured for another 24h. Then cells were refed with serum and further cultured. The results showed that protein B23 content was low in serum-starved NIH3T3 cells,and it markedly increased after serum addition. Furthermore,the increase occurred early at 6h after serum treatment. It was also showed that protein B23 content in mitotic cells blocked by the microtubule inhibitor colcemid was much higher than that in interphase ones. Our results suggested that nucleolar protein B23 might play some roles in the early stage of serum- induced proliferation and during transition from G2 to mitosis in cell cycle.

Objective To compare the early efficacy of two different ways of rehabilitation exercises in total knee arthroplasty (TKA) patients. Method From January 2011 to January 2014,136 patients having undergone TKA were randomly divided into two groups. In group A , patients exercised under the guidance with doctor. In group B, patients exercised by using continuous passive movement (CPM) machine. Compared the data from the two groups including range of motion (ROM) of the knee joint, knee function score (KFS) and circumference of the knee joint, t test of independent sample was utilized in the research. Results All patientswere followed up in 4 weeks. The ROM and KFS of Group A was significantly better than Group B (P < 0.05). The knee circumference had no statistically significant difference between two groups (P> 0.05).Conclusion Group A and B are effective methods in TKA patients in early rehabilitation training. But training under the guidance of doctor has greater advantages than CPM.

Objective To explore the mechanism of reversal of mutidrug resistance of GBC-SD cell lines by grape seed polyphenols(GSP).Methods GBC-SD cell lines were used to determine the effect of GSP.MTT assay was adopted to evaluate the cytotoxity(IC_(50)),RT-PCR were used to determine MDR1mRNA,(P-gp),bcl-2 and cellular adriamycin was measured by flow cytometry.Results In non-toxic(3?g/mL) and low toxic(6?g/mL) comcentration of GSP treated group(P

0.05) was observed. Bcl 2 expression was related with differentiation of GC(P0.05).Conclusion: p53,bcl 2,c erbB 2 may be involved in the development of GC.

Objective To study the effect of sub-glottal convergence angle on the degree of glottal closure, and to analyze the potential mechanism of dysphonia by the incomplete glottal closure.Methods Three vocal fold positions (adduction, intermediate and abduction) were evaluated by the degree of glottal closure, and divided into three groups.The neck CT images of the subjects were gathered.The vocal folds were adduction group when the normal subjects made a sustaining phonation /i:/ during CT scanning;the vocal folds were abduction group when the normal subjects made a deep breathing during CT scanning, and the unilateral vocal folds were incomplete closing group when the unilateral vocal fold paralysis subjects made a deep breathing during CT scanning.3D models of the vocal folds and the airway were reconstructed using Mimics software, through which the sub-glottal convergence angle was measured.Using one-way factor analysis of variance, we compared the angle among three groups.Results The angle changed with degree of glottal closure, with statistically significant differences among the three groups(P<0.01).The means of the angle in three groups were 33.49°±3.75°, 55.03°±2.61° and 75.02°±7.32°.Conclusion The 3D model generation from CT data is an effective method of measuring the sub-glottal convergence angle, while the angle changes with the degree of glottis closure.The angle may affect sub-glottal pressure distribution in the sub-glottal shear or normal direction and influence vocal fold vibration, which would make vibration and vocal fold mucosal wave generation more difficult, and might cause dysphonia.

Objective: To investigate the effect of survivin and bcl-2 antisense oligodeoxynucleotides ( AsODN) combined transfection on the human gallbladder carcinoma cell line GBC-SD in vitro. Methods: Survivin and Bcl-2 protein expressions were detected by immunohistochemical method; Cultured cells were divided into 4 groups: Nomal control group, survivin antisense observed group, bcl-2 antisense observed group and combined group. After transfected for 24 h, the expression of survivin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Cell morphological changes were observed under electron microscopy. Apoptosis index (AI) was examined by flow cytometry; Inhibitory rate (IR) was determined by the colorimetri MTT cell viability and proliferation assay. Results: Survivin and Bcl-2 protein were highly expressed in gall bladder carcinoma cells; The expression of survivin mRNA was decreased 47. 8%. Abnormal morphological changes of cells were observed in the three AsODN transfection groups; The AI in survivin antisense observed group,bcl-2 antisense observed group,and combined group was 11.38%?3.91% , 9.26%?4.15% , 28.45%?6.34% respectively and significantly higher than the nomal control group (P

Objective: To investigate effect of exogenous wild-type p53 gene on the cell growth and tumorigenicity of human gallbladder cancer cell lines. Methods: After identification of the genetic status of p53 gene of GBC-SD cell lines with the immunocytochemistry staining and the direct sequencing technique of PCR products, eukaryotic expressing plasmid pCMV-p53 was introduced by lipofectamine-mediated into GBC-SD cell lines. Growing transfected cells were selected by G418. The presence and expression of exogenous p53 gene was detected by PCR, RT-PCR and Western blot. The cellular proliferating ability was assessed using the cell growth curve and cloning assay. The xenograft in nude mice was performed to examine the effect of tumorigenicity. Results: P53 protein overexpression was showed in GBC-SD cell lines. A transversion of TAC→AAC at codon 126 of exon 5 was confirmed. PCR, RT-PCR and Western blot showed exogenous p53 gene had successfully transfected into GBC-SD cells and obtained high expression. The growth and proliferation of the cells were greatly decreased, and the tumorigenicity was significantly inhibited after transfection wtp53. Conclusion: The expression of exogenous wild-type p53 gene could effectively inhibit the growth of gallbladder cancer GBC-SD cells in vitro and in vivo.

Objective To study the apoptosis of gallbladder carcinoma cell line GBC-SD induced by antisense oligodeoxynucleotide (ASODN) targeting survivin. Methods ASODN targeting survivin was transfected into GBC-SD cells mediated by lipofectin. Cultured cells were divided into 3 groups: control group,sense oligonucleotide (SODN) group and ASODN group. After transfected for 16 h, the cultured cells were harvested and the following texts were carried out. The expression of survivin mRNA was detected by RT-PCR. Flow cytometer were used to detect apoptosis. Morphological changes were observed by electron microscopy. Results The expression of survivin mRNA was decreased 47.83% in ASODN group while apoptosis was increased from (0.50?0.23)% to (26.28? 3.91)%. Abnormal morphological changes of cells were observed in ASODN group and apoptosis bodies were found in some gallbladder carcinoma cells. Conclusion The expression of survivin may be decreased in GBC-SD cells after ASODN transfection.ASODN targeting survivin could induce gallbladder carcinoma cells apoptosis effectively.

Objective To investigate the effect of neonatal hypoxic myocardial injury and bradycardia on cerebral hemodynamics and brain tissue regional oxygen saturation (rSO2),and to provide patho-physiological evidence for treatment of neonatal hypoxic-ischemic encephalopathy (HIE) in acute phase.Methods Ninety nine full-term newborns admitted into Department of Neonatology,Peking University First Hospital from December,2005 to December,2008 were enrolled in this study.There were 18 newborns with both myocardial injury and HIE (group 1),31 newborns with HIE but without myocardial injury (group 2) and 50 cases of neonatal jaundice (control group).From 3 to 7 days after birth,Doppler ultrasound was used to measure systolic velocity (Vs),diastolic velocity (Vd) and resistance index (RI) of the anterior cerebral artery; and brain tissue rSO2 was detected by near infrared spectroscopy.The differences among groups were compared by variance analysis and LSD test.Brain tissue rSO2 before and after treatment of 20 newborns with bradycardia for various causes were detected and the difference was compared by paired t test.Results (1) Vs of group 1,group 2 and the control group was (19.35±5.13),(29.35±4.28) and (32.62±7.47) cm/s respectively; Vd was (6.43±2.98),(11.21±3.16) and (11.50±3.03) cm/s; RI was 0.68±0.10,0.62±0.03 and 0.64±0.06; brain tissue rSO2 was (52.4± 2.8)％,(54.6±3.1)％ and (62.1±1.9)％.There were significant differences among the three group(F=29.999,19.393,5.283 and 137.952,P＜0.01).Vs,Vd and rSO2 of group 1 were lower than that of group 2 and the control group (P＜0.05).RI of group 1 was higher than that of group 2 and the control group (P＜0.05).Vs and rSO2 of group 2 were lower than that of control group,but there were no differences between the two groups in Vd and RI.(2) The brain tissue rSO2 of newborns with bradycardia after treatment was higher than that before treatment [(58.7±4.6)％ vs (50.9±3.2)％,t=6.239,P＜0.01].Conclusions The disturbance of cerebral hemodynamics and cerebral oxygenation in HIE newborns are aggravated by hypoxic myocardial injury.Stable heart rate might be very important to early treatment for HIE.

Objective To investigate the correlation between hypoglycemia and brain injury of newborns.Methods Medical records and follow-up data of 110 newborns with hypoglycemia (blood glucose level≤2.2 mmol/L) who admitted into neonatal department of Peking University First Hospital from December 2006 to December 2009 were studied.All patients were divided into 3 groups:no brain injury group,mild and severe brain injury group according to their clinical manifestation,cerebral radiological characteristics and cerebral functional tests.By using receiver operating characteristic (ROC) curve and x2 test,the potential optimal blood glucose level and duration of hypoglycemia for predicting brain injury were confirmed.Multivariate Logistic regression was taken to determine independent predictors for brain injury.The analyzed factors included gender,preterm/small for gestational age,hyperbillirubinemia,fetal distress,asphyxia,infection,seizures and maternal hypertensive disorder complicating pregnancy and hyperglycemia.Results Among the 110 hypoglycemia newborns,33 (30.0％) infants suffered from brain injury,of which 23 were mild and 10 were severe.Blood glucose ≤1.7 mmol/L had high specificity (73％) and sensitivity (60％)for predicting brain injury.When blood glucose≤ 1.7 mmol/L,the incidence of brain injury and severe brain injury was 43.6％ (24/55) and 18.2％ (10/55),which was higher than those [16.4％(9/55) and 0.0％ (0/55)] of patients whose glucose level ＞1.7 mmol/L(x2 =9.74 and 11.00,P＜0.01 respectively).Blood glucose ≤ 1.2 mmol/L had high specificity (100％) and sensitivity (81％) for predicting severe brain injury.When blood glucose ≤1.2 mmol/L,the incidence of severe brain injury was higher than that of the patients whose glucose level was higher than 1.2 mmol/L [34.5％ (10/29) vs 0.0％ (0/81),x2 =30.72,P＜0.01].Duration of hypoglycemia ≥12 h had specificity (100％) and sensitivity (36 ％) for predicting brain injury.When duration of hypoglycemia ＜12 h,the incidence of brain injury was lower than that of the patients whose duration of hypoglycemia≥12 h [21.4％ (21/98) vs 6/6,x2 =27.69,P＜0.01].Multivariate Logistic regression showed that fetal distress (OR=4.69,95％CI:1.47-14.97,P=0.009),glucose level≤1.2 mmol/L (OR =5.16,95％CI:1.56-17.03,P=0.007),duration of hypoglycemia≥12 h (OR=8 885 220 297.12,95％CI:0.00-∞,P =0.000) and maternal hyperglycemia (OR =3.34,95％CI:1.01-11.02,P=0.048) were independent risk factors for neonatal brain injury.Conclusions Low blood glucose level and prolonged hypoglycemia might induce injury of neurol system.Fetal distress and maternal hyperglycemia might increase the incidence of brain injury in newborns with hypoglycemia.