A method was developed for the determination of ochratoxin A (OTA) in human urine by HPLC-FLD after molecularly imprinted polymer solid phase extraction (MIP-SPE) column. After the pH being adjusted to 2.5 with 0.1 mol x L(-1) HC1, sample was cleaned up with MIP-SPE column for ochratoxin A, the analyte was analyzed by high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD), and finally all the positive results were confirmed by LC-MS/MS. Recoveries from urine samples spiked with OTA at levels ranging from 2 to 20 ng x mL(-1) were 90.6%-101.9%, and RSDs were 0.1%-1.6%. Sixty-five volunteers living in Beijing took part in the study, of which 5 were found containing OTA in their urine and the highest value was 0.091 ng x mL(-1). The MIP-SPE column was firstly applied to purify and concentrate OTA in human urine, this method is simple, rapid and reliable and can be used to determine the contents of OTA in human urine.

The Epstein-Barr virus (EBV) is a gamma herpes virus associated with several types of malignancies. The EBV encodes viral microRNAs (miRNAs) that can target genes within cells. The EBV participates in signal transduction as well as the proliferation and differentiation of cells. How the target genes and functions of EBV-encoded miRNAs contribute to the pathogenesis of EBV is an important research topic. Some target genes have been validated since EBV-encoded miRNAs were discovered and, in this article, we summarize them and their functions.
Animals ; Epstein-Barr Virus Infections ; genetics ; metabolism ; virology ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; MicroRNAs ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism

Mycotoxins are secondary metabolites produced by certain genera of toxigenic fungus and frequently oc-cur in food worldwide. Humans and animals can be simultaneously exposed to different mycotoxins through the diet. As most mycotoxins are highly toxic, carcinogenic, teratogenic and mutagenic, they have posed grave health threats to consumers. Determination of mycotoxins and their main metabolites in blood, urine, bile, milk or faeces can serve as biomarkers and can facilitate effective exposure assessment, crucial to estimate mycotoxin related dis-ease risk. According to reason mentioned above, the study of metabolism and evaluations of mycotoxins in biologi-cal fluids have been paid increasing attention since the results may offer valuable indications on the real risk for consumers. Therefore, it is important to develop proper analytical methods for the rapid quantitative and qualita-tive measurement of mycotoxins and key metabolites in vivo. This paper reviewed some biomarkers and their harm to animals and humans, systematically summarized the research progress of analytical methods and prospected the development trends.

Objective To study the relationships between cell kinetics, apoptosis and expression, mutation of related genes in breast cancer. Methods Flow cytometry, immunohistochemical SP and PCR SSCP were used respectively to study DNA index(DI), S phase fraction(SPF), proliferation index(PI), apoptosis index(AI), expression of c erbB 2, Bcl 2, p53, PCNA, Ki67, TopoⅡ and mutation of p53 in 54 cases of breast cancer. Results High DI, aneuploidy rate, SPF, PI and AI were coincided with high expression of c erbB 2, p53, PCNA, Ki167 and TopoⅡ. Low DI, SPF, PI and AI were coincided with high expression of Bcl 2. High DI, SPF, PI and AI were coincided with high mutation of p53. High AI was related to a type of p53 mutation——loss of heterozygosity(LOH). Conclusion Abnormal cell kinetics and apoptosis were related with abnormal expression, mutation of related genes in breast cancer. [

Astrocytes were traditionally be deemed as supportive cells in the central nervous system.However,recent researches proved astrocytes exerted more important neurophysiological functions,such as regulation of synaptic transmission and integration of neural information.This paper summarized the researches on the functions of astrocytes related to synaptic transmission and analyzed the new features of astrocyte excitability,the communication between neurons and astrocytes and the effect of general anesthetics on astrocytes.Based on these new findings,this paper also suggested the underlying relationship between astrocytes and the loss of consciousness during general anesthesia.

AIM: To study the effects of paroxetine on the psychological stress induced by c-fos gene expression in the rat hypothalamus paraventricular nucleus(PVN) and to explore the molecular mechanism of effects of paroxetine on the stress related anxiety disorders. METHODS: The rat psychological stress model was made by restraint stress. The cortisol was analyzed by radioimmnoassay, and expression of c-fos positive cells was detected by S-P immunohistochemical assay in the rat PVN. RESULTS: The level of cortisol and the expression of c-fos positive cells increased more significantly in the other groups than that in the control group, but decreased in the paroxetine group. The paroxetine reduced the level of cortisol and inhibited the expression of c-fos positive cells in the PVN after psychological stress. CONCLUSION: The paroxetine can regulate the nerve centre by alleviating the expression of the c-fos in the hypothalamus paraventricular nucleus(PVN)and the activation of HPA pathway.

Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor on hypoxia responses in mammalian tissues. HIF-1 plays as a positive factor in solid tumor and leads to hypoxia-driven responses that enhance its downstream gene expression for tumor growth and survival. LXY6099 was obtained by the structural modification and optimization of manassantin A (MA) as a high potent HIF-1 inhibitor. Antitumor activity of LXY6099 was observed in this study. LXY6099 with an IC50 value of 2.46 x 10(-10) mol x L(-1) showed more sensitive inhibition activity to HIF-1 than that of MA detected by reporter gene assay (> 100 folds). It showed strong inhibition on the growth of human solid tumor cell lines. Furthermore, LXY6099 exhibited significant antitumor activity against established human tumor xenografts in nu/nu mice with treatment of MX-1 breast cancer. Thus, LXY6099 as a novel HIF-1 inhibitor could be further developed into anti-cancer agents.

Objective To study Kaposi's sarcoma-associated herpesvirus(KSHV)infection in chronic hepatitis B(CHB)patients and its correlation with hepatitis B virus(HBV)replication and treatment-related factors.MethodsEnzyme-linked immunosorbent assay(ELISA)with recombination protein KSHV ORF65 was employed to detect the KSHV antibody and real-time polymerase chain reaction(PCR)was performed to detect KSHV DNA and HBV DNA in CHB patients.Age,HBV replication and licorice preparation treatment of patients were further analyzed.Comparison of rates was done using X~2 test.Results KSHV ORF65 antibody positive rates were 27.3% in 161 male CHB patients and 30.0% in 50 female patients(X~2=0.135,P＞0.05).The KSHV infection rates were increased with age,but this tendency was not obvious in patients older than 40 years old.The highest infection rate was in age group of 31-40 years old which was 37.1%.The positive rate of HBV DNA in CHB patients with KSHV infection was 73.5%,which was 56.3% in uninfected patients(X~2=3.969,P＜0.05).The average plasma level of KSHV DNA in patients treated with licorice preparations was 204.7 copy/mL and that in patients without licorice preparation treatment was 533.9 copy/mL.Eight patients were KSHV DNA positive(KSHV DNA＞ 100 copy/mL)in 16 patients treated with licorice preparations and 23 were positive in 33 patients without licorice preparation treatment.Conclusions The KSHV infection rates are increased with age of CHB patients.KSHV infection may interfere with HBV replication and licorice preparations may suppresss KSHV replication in vivo.

Objective To investigate the effect of APE1 on differentiation of peripheral blood mononuclear cells into osteoclast-like cells(OCL) which induced by macrophage colony stimulating factor (RANKL) and macrophage colony stimulating factor (M-CSF) .Methods Human peripheral blood mononuclear cells (PBMCs) were collected by density gradient separation ;Constructed APE1 siRNA expression vector Ad5v-APE1 siRNA was used to transfect PBMCs .Tartrate-resistant acid phosphatase (TRAP) method was conducted to identify the cells ,the expression level of APE1 was detected by Western blot ,the mRNA expression levels of Cathepsin K(CK) and V-ATPase were detected by RT-PCR .Results PBMCs transfected with APE1 siRNA had significantly lower protein expression of APE1 than untransfected cells (P< 0 .05) ;PBMCs could differentiate into OCL under the stimulation of RANKL and M-CSF ,the mRNA expression levels of CK and V-ATPase increased ;After APE1 siRNA treatment ,the number of OCL was reduced and the levels of CK and V-ATPase mRNA decreased .Conclusion PBMCs can differentiate into a large number of OCL induced by RANKL and M-CSF ,APE1 siRNA significantly inhibited differentiation of PBMCs into osteoclast-like cells , APE1 may be involved in the regulation of osteoclast-like differentiation process .

Objective To find the possible pathogenesis of enteric nervous system, gut hormone and gastric Cajal interstitial cell ( ICC ) in gastritis related gastrointestinal ( GI ) motor disorders on the changes of protein gene product 9. 5 in neurons , mucosal expression of C-kit, gastrin and somatostatin from the gastric wall of gastritis rat. Methods 45 rats were divided into 3 groups which included gastritis group A, gastritis group B and control group. Rats in gastritis group A were fed with Hp Sydney Strain 1, the mixture of 2% aspirin and 0. 6N hydrochloric acid was fed in gastritis group B. The control group only received saline. All of the rats were killed and mucosal tissue was obtained from antrum and greater curvature of the gastric body. Pathological and Hp examination were performed in the tissue slides, and then it was stained to check PGP 9. 5, gastric body's mucosal expression of C-kit, antrium's mucosal expression of gastrin and somatostatin. The cell body, the maximum diameter (Dmax, μm), mean area( μm2) and optical density (nm), integral optical density of the gastrin and somatostatin in the C-kit expression positive neurons from the gastric wall were compared among the groups. Result The mean area and optical density of PGP 9. 5 expression in neurons from the gastric wall of rat in group A or B were obviously lower than that of the control group ( P ＜0. 01 ), while there was no difference between gastric group A and B ( P ＞0. 05). Gastric group A had higher GAS expression than control group, while SS expression was lower than control group( P＜0. 05). There was no difference between group B and the control group in the two variances( P ＞0. 05).By linear correlation analysis, it showed that SS was negatively correlated with GAS ( r = - 0. 333, P ＜0. 01 ). The distributive area and diameter of cells with C-kit expression in both group A and B were significantly smaller than that in the control group ( P ＜ 0. 05 ), while there was no obvious difference between group A and B ( P ＞ 0. 05 ). There was no difference of integral optical density of the C-kit expression positive neurons among the three groups. Conclusions Hp infection and NSAIDs might cause gastritis and had influence on the structural changes of neurons from gastric wall and ICC. Hp infection could obviously inhibit SS excretion from antrum mucosa while increase Gastrin excretion. NSAIDs induced gastritis had little influence on GAS and SS.