Objective To investigate the inhibitory effect of RNA interference(RNAi)on WT1 gene expression in HL-60 cell line.Methods Several pairs of small interfering RNA(siRNA)for WT1 gene was designed and transfected into HL-60 cells.WT1 gene expression was assessed by RT-PCR,and WT1 protein expression was tested by Western blot.Flow cytometry was used to measure the cell apoptosis.Results The expression rates of WT1 mRNA and the expression rates of WT1 protein were(23.62%?1.28%),(23.20%?1.04%),respectively in HL-60 WT1 siRNA-transfected cells,significantly lower than those of control cells(P

Autoantibodies ; Child ; Complement Factor H ; immunology ; Female ; Hemolytic-Uremic Syndrome ; immunology ; Humans

Objective To investigate the risk factors for hip fracture in elderly people in Ningbo,in order to provide the basis for prevention and treatment of hip fracture.Methods A case-control study were conducted in study on 450 cases of hip fracture patients aged over 60 years in Ningbo and 450 subjects of control as 1 ∶ 1 matched case control study.A univariate analysis of hip fracture was performed by x2 test.Risk factors for hip fracture were analyzed by logistic regression.Results Poor self-care ability (OR=5.05),light manual labor occupation (OR=4.10),history of cerebrovascular events (OR=2.59),smoking (OR=2.14) were the risk factors for hip fracture in elderly men,while poor self-care ability (OR=11.36),light manual labor occupation (OR=2.41),osteoporosis (OR=3.19),early menopause (OR=6.89),low body mass index (OR=5.05) were the risk factors for hip fracture in elderly women.The protective factors for hip fracture was milk uptake (OR=0.29) Conclusions Early interventions for hip fracture risk factors,and prevention and treatment of osteoporosis can help reduce the occurrence of hip fracture.

Objective To analyze the effects of curcumin combined with cisplatin on the growth and lymphatic metastasis of cervical cancer xenografts in nude mice.Methods The human cervical carcinoma Caski cells xenotransplanted tumor models were established.Inoculated mice were randomly divided into 4 groups according to random number table:control (normal saline 10 ml/kg),Cur (curcumin 100 mg/kg),Cis (cisplatin 3 mg/kg) and Cur + Cis group (100 mg/kg curcumin +3 mg/kg cisplatin).The tumor volume and anti-tumor rate were calculated.The mRNA levels of macrophage migration inhibitory factor (MIF) and vascular endothelial growth factor-C (VEGF-C) were analyzed by real-time fluorescent quantitative polymerase chain reaction (PCR).Results The inhibitory rates of Cur group,Cis group and Cur + Cis group were 40.8％,53.3％ and 60.0％.After 15 days treatment,the tumor volumes of the control group,Cur group,Cis group and Cur + Cis group were (123.44 ± 35.62),(71.72 ± 28.36),(65.47 ± 18.32),(53.44 ± 10.79) mm3.The growth rates of tumor volume in Cur group,Cis group and Cur + Cis group were slower,Cur + Cis group could more effectively inhibit tumor volume growth.The difference among the four groups had statistically significance (F =16.890,P =0.000).Real-time fluorescent quantitative PCR detection showed that compared with the control group (1.000),the MIF mRNA expressions in Cur group,Cis group and Cur + Cis group were 0.322 ±0.094,0.154 ± 0.006 and 0.136 ± 0.007,VEGF-C mRNA expressions in the three groups were 0.312 ± 0.068,0.263 ± 0.072 and 0.221 ± 0.041.The differences among groups had statistically significance (F =220.279,P =0.000;F =143.250,P =0.000).MIF mRNA was positively related with VEGF-C mRNA (r =0.815,P =0.001).Conclusion Curcumin combined with cisplatin can inhibit the growth of Carski cell xenografts in nude mice.Through the down-regulating expression of MIF mRNA and VEGF-C mRNA,cisplatin and curcumin can inhibit the lymphatic metastasis of cervical cancer,and it may be one of the important mechanisms of its anti-tumor effects.

Objective To explore the effect of different type of atrial fibrillation(AF) on plasma brain natriuretic peptide(BNP) level in male patients.Methods Fifty-eight hospitalized male patients with AF were enrolled,including 35 cases with paroxysmal AF,14 cases with persistent AF,and 9 cases with permanent AF.All subjects had the examination of echocardiography/electrocardiogram within 24 h after admission.Venous blood sample were collected in a quiet prostration state on the next early morning for BNP detection.The plasma BNP level was determined by radioimmunology-assay(RIA).Results The average level of plasma BNP in male patients with permanent AF(2343.25 ± 820.24)μg/L was significantly higher than those with persistent AF([576.43 ± 223.07]μg/L,t=8.74,P＜0.001) and paroxysmal AF([632.74 ± 103.93]μg/L,t=8.73,P＜0.001),but there was no significant difference between the last two groups(t=0.59,P＞0.05).Conclusion BNP is a sensitive indicator of early heart function impairment related with AF,particularly elevates in male patients with permanent AF.

Objective To investigate the effect of lipopolysaccharide (LPS) on stimulating high mobility group box-1 protein (HMGB1) mRNA expression in peritoneal macrophages in rats and its potential signaling mechanism. Methods Abdominal macrophages obtained from male Wistar rats were seeded on 24-well (1?10 6 cells/well) tissue culture plates. The cells were incubated for 3 days before they were stimulated with LPS, fludarabine (specific inhibitor of signal transducer and activator of transcription 1, STAT1), or rapamycin (specific inhibitor of signal transducer and activator of transcription 3, STAT3). After being stimulated, macrophages were denatured directly in tissue culture plates to determine HMGB1 mRNA expression. The time-dependent and dose-dependent responses between LPS stimulation and HMGB1 mRNA expression were analyzed, and the effect of treatment with fludarabine and rapamycin on HMGB1 mRNA expression was also observed. Results After being stimulated with 75-100ng/ml LPS, the HMGB1 mRNA expressions in macrophages were up-regulated markedly, peaking at 24-36 hours, and remained elevated up to 48 hours. It was found that the HMGB1 mRNA expression was significantly inhibited by treatment with either fludarabine (100?mol/L) or rapamycin (25ng/ml). Conclusions These data suggest that LPS stimulation can result in up-regulation of HMGB1 mRNA expression in peritoneal macrophages in rats. Janus kinase/STAT pathway may be involved in modulating HMGB1 mRNA expression in LPS-activated macrophages.

Background The visual evoked potential (VEP) is an important functional index to assess visual cortex plasticity.Monocular form deprivation(MD) of rat is a classic model to study visual cortex plasticity change.Utilizing VEP technique record the shift of ocular dominance in adult rats model is of an important significance for explore the treating opportunities of amblyopia.Objective The present study was to observe the pattern VEP changes in different ages of Long-Evens rats and the adult rats after binocular form deprivation.Methods Thirty-six healthy SPF Long-Evens rats were divided into PW3,PW4,PW5,PW6 groups according to the postnatal weeks,and PW7 rats included 36 SPF Long-Evens rats.The left eye lids of the rats were sutured in PW3,PW6 and PW7 rats for 3,5,7 days respectively,and PVEP in both left and right eye were recorded to assess the rat age that visual cortex plasticity ended.Bilateral PVEP were recorded in PW7 rats,and the bilateral eyelids were sutured to establish the bilateral form deprivation models.The right eyes were opened in 7,10,14 days,and the left eyes were opened in the next 3,5,7 days respectively and the PVEPs were recorded again to find the shift of ocular dominance and whether binocular form deprivation induce the visual cortex plasticity in adult rats.Results The P100amplitudes of the left eyes were gradually declined and those of the right eyes were raised from 3 to 7 days after MD of the left eyes in rats of PW3 group in comparison with before MD ( P＜0.01 ).In PW6 groups,no significant changes in P100amplitudes of both the right and left eyes were found in the third day after MD,but significant raise in the right eyes and lowing in the left eyes were seen in the 5 and 7 days after MD in comparison with before MD(P＜0.01 ).No any P100changes in both eyes were found in 3-7 days after MD compared with before MD in PW7 rats (P＞0.05).The C/I ratio (contralateral VEP amplitude to ipsilateral VEP amplitude in occluded eyes) in the rats of PW3,PW4 and PW5 groups were 1.07±0.15,1.16±0.16 and 1.14±0.15 respectively in 3 days after MD,showing the significant lowing in comparison with before MD (2.69±0.45,2.58±0.4,2.62±0.32) (P＜0.01),but those of PW6 and PW7 were unchanged (2.80±0.48 vs 2.90±0.46,2.59 ±0.36 vs 2.90±0.46,P＞0.05 ),indicating the absence of ocular dominance plasticity in the adult rats.In rats of PW7 with binocular form deprivation for 14 days,a significant decrease was observed in the C/I ratio in rats with next MD for 3 days,demonstrating that the visual cortex plasticity was reactivated after 14 days of binocular form deprivation in PW7 rats( 1.33±0.18 vs 2.70±0.45,P＜0.01 ).Conclusions PVEP can be recorded in Long-Evens rats.It is a major index for identifying the shift of ocular dominance in the Long-Evens rats.Binocular form deprivation can reflect the visual cortex plasticity in the adult rats.

Objective To explore the change of plasma D-dimer (DD),lactate dehydrogenase isoenzyme 3 (LDH3),tissue plasminogen activator (t-PA) in the early stage of acute pulmonary embolism and their clinical significance.Methods Forty-six patients with acute pulmonary embolism of early stage (pulmonary embolism group) and 50 cases of normal person (control group) were enrolled in this study.The patients in pulmonary embolism group were divided into low-risk(18 patients),middle-risk(15 patients) and higher-risk (13 patients).The levels of DD,LDH3 and t-PA were detected and compared.Results The levels of DD,LDH3 and t-PA in pulmonary embolism group were (2.95 ± 0.73) mg/L,(421.42 ± 63.29)U/L and (0.85 ±0.02) U/L,and significantly higher than those in control group [(0.03 ±0.01) mg/L,(198.17 ± 23.37) U/L and (0.59 ± 0.02) U/L] (P ＜ 0.05).The levels of DD,LDH3 and t-PA in low-risk patients [(1.49 ± 0.04) mg/L,(315.47 ± 38.24) U/L and (0.63 ± 0.01) U/L] were significantly lower than those in middle-risk patients [(2.22 ±0.27) mg/L,(382.41 ±54.36) U/L and (0.79 ±0.02) U/L] and higher-risk patients [(3.38 ± 0.98) mg/L,(583.93 ± 117.21) U/L and (0.92 ± 0.03) U/L] (P ＜ 0.05).The levels of LDH3 and t-PA in middle-risk patients were significantly lower than those in higher-risk patients (P ＜ 0.05).After treatment,the levels of DD,LDH3 and t-PA were significantly decreased [(2.95 ± 0.73)mg/L vs.(1.03 ±0.11) mg/L,(421.42 ±63.29) U/L vs.(223.24 ±25.04) U/L,(0.85 ±0.02) U/L vs.(0.62 ± 0.02) U/L] (P ＜ 0.05).Conclusions The levels of DD,LDH3 and t-PA in the early stage of acute pulmonary embolism are increased significantly,and with the remission of the patient's condition,the levels of DD,LDH3 and t-PA are decreased.United detection conduces to the diagnosis of acute pulmonary embolism.

Objective To explore the effect of rhynchophylline and isorhynchophylline on headshakes behavior and levels of monoamine neurotransmitter in model rats with Tourette syndrome.Methods 40 Wistar rats were randomly divided into DOI-induced head-shakes rats (HSR group),haloperidol group,rhynchophylline group and isorhynchophylline group with 10 in each group.The inhibitory effects of rhynchophylline and isorhynchophylline were estimated by observing the HSR behavior.Dopamine (DA) and 5-hydroxytryptamine(5-HT) in the rat striatum were detected by the enzyme-linked immunosorbent assay (ELISA) method.The 5-HT2A receptor mRNA expression in prefrontal lobe cortex of the rats was measured by real-time PCR.Results Compared with HSR group,the head shakes of the rats in haloperidol group and isorhynchophylline group were significantly decreased(P＜0.01),and no change of head-shakes number was observed in rhynchophylline group (P＞0.05).There was no significant difference of head-shakes number between the haloperidol group and isorhynchophylline group(P＞0.05).Compared with HSR group,DA levels in the rat striatum were significantly decreased in isorhynchophylline group and haloperidol group((152.35± 5.80) μ~L vs (111.19±4.30) μg/L,(152.35±5.80) μg/L vs (126.42±3.17) μg/L,P＜0.01),while DA levels in the rat striatum in rhynchophylline group were not changed ((152.35±5.80) μg/L vs (142.71±5.51) μg/L,P＞0.05).There was no significant change of 5-HT2A receptor mRNA expression in rat prefrontal lobe cortex in every group(P＞0.05).Conclusion Isorhynchophylline may have an inhibitory effect on rats with DOI-induced HSR.Isorhynchophylline may decrease the DA levels in the rat stratum with DOI-induced HSR.Rhynchophylline has no significant inhibitory effect on head-shakes behavior and DA levels in the rat stratum with DOI-induced HSR.

Objective To explore the mechanism of ischemic postconditioning in relieving cerebral ischemia reperfusion (IR) by regulating autophagy through P38MAPK pathway.Methods Cerebral ischemia reperfusion model was established by using modified Pulsinelli four-vessel occlusion (4-VO).Totally 128 male SD rats were divided into 4 groups randomly:control group (sham),cerebral ischemia reperfusion model group (CIR),cerebral ischemic postconditioning group (CIP),and cerebral ischemic postconditioning + P38MAPK inhibitor group (SB203580 group).Each group was subdivided into four time points:6 h,24 h,48 h,and 72 h.The morphological changes of the hippocampus CA1 area neurons at each time point and the number of surviving nerve cells were detected with HE staining.The expression of the hippocampus CA1 area phosphorylated P38MAPK and the autophagy-related genes of Beclin-1 and LC3-Ⅱ were detected with immunohistochemistry.The protein content of the hippocampus phosphorylated P38MAPK and autophagy-related genes of Beclin-1 and LC3-Ⅱ were detected with Western blotting.Results Compared with those in sham group,the damage of rats' hippocampal neuron structure and the survival rate of neurons at each time point decreased in CIR group,the expressions of p-P38MAPK,LC3-Ⅱ and Beclin-1 increased.Compared with those in CIR group,in CIP and SB203580 groups the structure of rats hippocampal neurons was improved,the survival rate of neurons increased,the expression of p-P38MAPK decreased and the expressions of LC3-Ⅱ and Beclin-1 increased at each time point.Compared with CIP group,SB203580 grouphad improved structure of rats' hippocampal neurons,increased survival rate of neurons,decreased expression of p-P38MAPK,and increased expressions of LC3-Ⅱ and Beclin-1 at each time point.Conclusion Cerebral ischemic postconditioning through inhibiting P38MAPK pathway can regulate autophagy and exert its nerve-protective effect.