Objective To study the effects of extract from Narcissus tazetta var. chinensis on proliferation and apoptosis of human multiple myeloma cell line ARH-77. Methods The survival rate of cells were tested by MTT assay when cells were affected by 1.25, 2.5, 5, 10, 20 ?g/mL of the extract for 3 d. After being induced by 10 ?g/mL of the extract for 3 d, apoptotic cells were observed with fluorescence stain; changes of he csll cycle were analyzed by Flow Cytometry; the number and shape of nucleolar organizer region (NOR) were tested by AgNOR assay and submicroscopic structure were observed by transmission electron microscope. Results The extract from N. tazetta var. chinensis could significantly inhibit the proliferation and reduce the survival rate of ARH-77 cells (P

BACKGROUND: Hippocampal neuron presents remarkably injury in cerebral after seizure of epilepsy. Necrosis and apoptosis are two kinds of neural cell injury after epilepsy and play an important role in neural injury of epilepsy. Being endogenous neural protective transmitter, adenosine may inhibit the release of excitatory amino acid, production of oxygenic free radical and action of nitric oxide. Simultaneously, it can improve cerebral blood flow and anti-convulsion. But it has been unknown concerning to the relationship between adenosine and cell apoptosis after epilepsy yet.OBJECTIVE: To observe the effects of 2-CAdo adenosine-receptor excitant on genetic expression of bcl-2, Bax of hippocampal cells in epileptic rats and further probe into the mechanism of adenosine on anti-convulsion and brain protection.DESIGN: Completely randomized controlled experimental research in which the experimental animals were taken as the objects.SETTING: Pediatrics department and general surgical department of one oil field general hospital, and pediatric internal department of a hospital affiliated to one university.MATERIALS: The experiment was performed in Experimental Zoology Departnent and Pathological Teaching & Research Department of Harbin Medical University from October 2002 to March 2003. Totally 104 Wistar rats of either sex were employed, weighing varied from 200 g to 250 g. The animals were randomly divided, named as normal group 8 rats, epileptic group 32 rats, epileptic & 2-CAdo group 32 rats, and epileptic & physiological saline group 32 rats.INTERVENTIONS: The animal epileptic model was set up by intra-abdominal injection of coriamyrtin 15 mg/kg(provided by Pathology Department of Harbin Medical University. Convulsion presented in all of rats, 5 minutes later after injection, lasting for 1 or 2 minutes. In epileptic & 2-CAdo group, 2-CAdo(provided by ICN company), 0.6 mg/kg, was injected from the vein on the tail 1 hour before coriamyrtin injection and 1 hour after convulsion respectively. In epileptic & physiological saline group, the physiological saline of equal dosage was injected from the vein on the tail 1 hour before coriamyrtin injection and 1 hour after convulsion respectively.MAIN OUTCOME MEASURES: Positive cell counts of bcl-2 and Bax genetic expression in hippocampal CA1 area.RESULTS: Twenty-four hours after epilepsy seizure, neural cell bcl-2 expression was increased in hippocampal CA1 area, was remarkably decreased in 48 hours, and the expression was only little amount in 72 hours, but it was increased again in 7 days. Bax expression began increased in 24 hours after epilepsy seizure, was significantly increased in 48 hours, reached the peak in 72 hours, the expression was the minimum in 7 days. In epileptic & 2-CAdo group, bcl-2 expressions at corresponding times were remarkably increased compared with epileptic group and epileptic & physiological saline group( P＜ 0.05), Bax expressions were remarkably decreased compared with epileptic group and epileptic & physiological saline group( P ＜ 0.05), indicating statistical significance.CONCLUSION: 2-CAdo can reduce apoptosis of hippoeampal neural cells after epilepsy seizure and provide a certain protection for neural cells.

Objective To evaluate the level of MMP-2 and COX-2 Protein in bladder transitional cell carcinoma tissue and explore their relationships. Methods A total of 42 patients with bladder transitional cell carcinoma, including Ta-T1 (n=18), T2-T4 (n=24), G1(n=12), G2 (n=19), G3 (n=11), metastasis (n=26) and without metastasis (n=16), were enrolled in the study. Eight normal bladder tissues were selected as control group. Western blotting was performed todetect the mRNA level of MMP-2 and COX-2. Results The relative COX-2 protein level of Ta-T1 (0.729±0.458), T2-T4 (1.248±0.425), G1 (0.61±0.486), G2 (1.055±0.406), G3 (1.422±0.341) were all higher than that of the control group significantly (0.31±0.149, t = 3.56, 4.13; F = 5.98, P ＜0.05). The relative MMP-2 protein level of Ta-T1 (0.844±0.345), T2-T4 (1.458±0.463), G1 (0.971 ±0.370), G2(1.445±0.378), G3 (1.755±0.387) were all higher than that of the control group (0.460±0.213, t = 3.91, 4.83;F = 6.35, P ＜0.05). The COX-2 and MMP-2 protein level in tumor tissues with and without metastasis were 1.246±0.426 vs 0.668±0.421, 1.430±0.461 vs 0.814±0.341, t = 5.89, 6.27, P ＜0.01, respectively. The level of COX-2 protein was positively correlated with MMP-2 positively (r =0.48, P ＜0.01). Conclusion MMP-2 and COX-2 protein are highly expressed in bladder transitional cell carcinoma tissue and their expression is positively correlated with the malignant degree. MMP-2 and COX-2 might play a synergetic role in the carcinogenesis of bladder transitional cell carcinoma.

Objective To investigate the effects of arsenic trioxide (AS2O3)on SOCS-1 gene methylation and expression of P-STAT3 in multiple myeloma (MM) cells.Methods MM cell lines U266 and CZ-1 were used as in vitro models.Methylation status of SOCS-1 gene was detected by the methylation specific PCR (MSP)while P-STAT3 protein expression was determined by Western blotting assay before and after AS2O3 treatment.Meanwhile growth inhibition and apoptosis of MM cells were determined by flow cytometry.Results Hypermethylation of SOCS-1 gene was observed in each MM cell line compared with wide type.After exposure to AS2O3,it was shown that SOCS-1 gene was demethylated obviously,meanwhile the expression level of P-STAT3 protein and cell proliferation was inhibited significantly in each cell line.The apoptosis rate was increased.When U266 and CZ-1 were treated with AS2O3 of 0,0.5,1.0,2.0 μmol/L respectively,the total cell apoptosisis ratio of U266 was 0.06％,0.56％,48.96％,61.07％ (X2 =9.19,P ＜ 0.05); and the total cell apoptosisis ratio of CZ-1 was 4.2％,,40.3％,,47.72％,,68.49％ (X2 =8.96,P ＜0.05 ).Conclusion AS2O3 could inhibit JAK/STAT signal transduction pathway by inducing SOCS-1 gene demethylation in MM cells which might be related to cell apoptosis.

Objective To screen genes differentially expressed between dermal papilla cells from occipital and vertex scalp of patients with androgenic alopecia (AGA) through a 3D culture system.Methods Dermal papilla cells isolated from the occipital scalp tissue of patients with AGA were cultured in a 2D system for several days.Then,the third-passage dermal papilla cells were subjected to a 3D culture with the presence of dihydrotestosterone (DHT) for 72 hours (experimental group).The dermal papilla cells isolated from the vertex scalp tissue of patients with AGA,which were cultured in a 3D system with dimethyl sulfoxide,but not DHT,served as the control group.Subsequently,total RNA was extracted from the cells and reversely transcribed into cDNA followed by labeling with Cy3 and hybridization to a NimbleGen microarray.Gene ontology (GO) and pathway analysis was carried out to screen differentially expressed genes between the experimental and control group.Real time PCR was conducted to validate the results of microarray analysis.Results As the genome-wide expression profile analysis showed,there were 622 genes differentially expressed between the experimental group and control group,of which,359 were up-regulated and 263 were down-regulated in the experimental group compared with the control group.The above results were corffirmed by real time PCR.GO analysis revealed that the up-regulated genes,such as the CHEK1 and Tobl genes,were mainly involved in the inhibition of cell proliferation and promotion of cell apoptosis,while the down-regulated genes,such as the BAMBI,EFNA3,Dlx3 and UCGC genes,were associated with the acceleration of cell proliferation as well as the growth and development of epidermis.Pathway analysis showed that cell circle-controlling molecules were the most abundant molecules.Conclusions Numerous signalling molecules and pathways are involved in the development of AGA,which are mainly responsible for the modulation of cell circle,proliferation and apoptosis.

ObjectiveTo investigate the difference in the effects of sevoflurane inhalation on hippocampal neuronal phosphorylated cAMP response element-binding protein between male and female rats.MethodsFiftyeight healthy SD rats aged 3 months weighing 180-440 g were randomly divided into 4 groups:group female control (Fc group,n ＝ 15) ; group female sevoflurane (Fs group,n ＝ 15) ; group male control (Mc group,n ＝ 14) and group male sevoflurane (Ms group,n ＝ 14).The 2 control groups (Fc group,Mc group) inhaled 95％ 02 for 2 h,while the 2 sevoflurane groups (Fs group,Ms group) inhaled 3％ sevoflurane for 2 h.The cognitive function was assessed by passive avoidance task performed on the 2nd day after sevoflurane inhalation and Morris water maze test once a day for 5 consecutive days from day 3-7 after sevoflurane.The animals were sacrificed after last cognitive function assessment test on the 7th day after sevoflurane inhalation and their brains were removed for determination of expression of hippocampal neuronal p-CREB1,Bcl-2 and caspase-8 protein expression.ResultsSevoflurane inhalation significantly increased the escape latency and swimming distance at day 3 after sevoflurane inhalation in group Fs and at days 3-6 in group Ms as compared with their control groups (Fc group,Mc group) in Morris water maze test.The escape latency and swimming distance were significantly longer at 4-6 d in Ms group than in Fs group.Sevoflurane significantly decreased p-CREB1 and Bcl-2 protein expression and increased caspase-8 expression in groups Fs and Ms as compared with their control groups (Fc group,Mc group).Bcl-2 protein expression was significantly higher in group Fs than in group Ms.ConclusionTwo hour 3 ％ sevoflurane inhalation can induce hippocampal neuronal apoptosis by down-regulating CREB1 phosphorylation and Bcl-2 expression and up-regulating caspase-8 expression.The effects are greater in male rats than in female rats.

Objective To explore the possible relationship between alteration of cell cycle and JAK/STAT3 signal transduction pathway inhibition induced by arsenic trioxide (As_2O_3,) in myeloma cell lines U266 and RPMI8226 in vitro. Methods Multiple myeloma cell lines U266 and RPMI8226 were used as in vitro models. The influence of AS_2O_3 on myeloma cells were evaluated by MTT assay and flow cytometry. Meanwhile, methylation specific PCR and Western blotting were employed to detect the methylation status of gene SOCS-1 and protein expression level of P-STAT3 in these cells after AS_2O_3 treatment. Results AS_2O_3 significantly inhibited the growth of U266 and RPMI8226 cells in a dose-dependent manner. Furthermore, cell cycle was arrested at G0/G1 phase with inhibition of protein expression level of P-STAT3 and SOCS-1 gene demethylation after exposure to As_2O_3 for 72 h( r = 0. 85, P < 0.05). Conclusion AS_2O_3 could induce the alteration of cell cycle which might be related to JAK/STAT3 signal transduction pathway inhibition and SOCS-1 demethylation in myeloma cell lines. The study puts forward a new idea of AS_2 O_3 treatment in multiple myeloma.

Objective To investigate the clinical value of miRNA-34a,miRNA-34c,and miRNA-135a in the early diagnosis of mild cognitive impairment (MCI) and Alzheimer's disease (AD).Methods The patients were collected in the outpatient and inpatient of Neurology during Aug 2014 to May 2016.The levels of miRNA-34a,miRNA-34c,an dmiRNA-135a were detected with quantitative real-time polymerase chain reaction (qRT-PCR).Results AD patients had higher level of miRNA-34a than the controls.The levels of miRNA-34c were higher in MCI and AD patients,while lower levels of miRNA-135a compared to the controls.Conclusions The miRNA 34a and miRNA-135a were likely to became the biomarker in early diagnosis of MCI and AD.

In order to investigate peptide mimics of carbohydrate blood group A antigen, a phage display 12-mer peptide library was screened with a monoclonal antibody against blood group A antigen, NaM87-1F6. The antibody-binding properties of the selected phage peptides were evaluated by phage ELISA and phage capture assay. The peptides were co-expressed as glutathione S-transferase (GST) fusion proteins. RBC agglutination inhibition assay was performed to assess the natural blood group A antigen-mimicking ability of the fusion proteins. The results showed that seven phage clones selected bound to NaM87-1F6 specifically, among which, 6 clones bore the same peptide sequence, EYWYCGMNRTGC and another harbored a different one QIWYERTLPFTF. The two peptides were successfully expressed at the N terminal of GST protein. Both of the fusion proteins inhibited the RBC agglutination mediated by anti-A serum in a concentration-dependent manner. These results suggested that the fusion proteins based on the selected peptides could mimic the blood group A antigen and might be used as anti-A antibody-adsorbing materials when immunoabsorption was applied in ABO incompatible transplantation.
Adsorption ; Bacteriophages ; Blood Group Antigens/*chemistry ; Enzyme-Linked Immunosorbent Assay ; Epitopes/chemistry ; Glutathione Transferase/metabolism ; Peptide Library ; Peptides/*chemistry ; Protein Structure, Tertiary ; Recombinant Fusion Proteins/chemistry

Objective To learn 2009~2014 Shaanxi Province sentinel surveillance six classes of HIV infection focus groups, and estimates of HIV-1 new infection.Methods Used enzyme-linked immunosorbent assay (ELISA)and Western blot (WB)experiments for the 2009~2014 HIV sentinel surveillance Shaanxi Province Category 6 focus groups conducted a total of 77 778 HIV antibody screening and confirmatory testing estimates of HIV-1 new infection.Results 2009~2014 men who had sex with men and people with HIV infection rate were 3.75%,8.77%,3.50%,5.00%,6.20% and 5.75%,and a slow upward trend;HIV-1 new infection were 5.04%,8.96%,5.01%,5.95%,4.68% and 6.39%,the overall downward trend. Young students,drug addicts,sex workers,pregnant women,and male STD clinic attenders five people with HIV infection and HIV-1 new infection were emerging to remain low.But male STD clinic attenders of HIV infection and HIV-1 new in-fection was emerging slowly rising trend.Conclusion Shaanxi MSM HIV infection and HIV-1 new infection were high,but HIV-1 new infection had decreased slowly.Emerging trend should continue to increase the population of the intervention ef-forts.The infection rate in other monitoring population was relatively low but a few people on the rise,the need to take the necessary coping methods.