Circadian rhythms are endogenous 24 h variations found in virtually all physiological processes and behaviors, which are controlled by the transcriptional translational oscillator that consists of a series of core clock genes (bmal1, clock, cry and per) and clock controlled genes (rev-erbα, rorα, dbp, tef and hlf).Clock genes exist in immune organs, tissues and cells, leading to the immune cell function (migration and chemotaxis, phagocytosis, cytotoxicity and so on) and a variety of immune parameters (factor level of circulating immune cells and subsets of the relative and absolute number of cells) showing circadian rhythm changes, and playing an important role in maintaining immune homeostasis.In addition, some immune related diseases are closely correlated with circadian rhythms abnormalities.This paper will focus on the effect of circadian rhythms on immune functions and their roles in some immune related diseases.

Objective　To investigate the IL-6 signal transduction pathways and their regulation mechanism in a human myeloma cell line-U266. Methods　Electrophoretic mobility shift assay (EMSA) was used to detect the activation of the transcription factors (TFs)-STAT3 and NF-IL-6 by IL-6. Cells were treated with chemical agents or transfected with the expression plasmids for the two TFs or the anti-sense oligonucleotide for protein kinase involved in one IL-6 signal transduction pathway. The change of the activation state of another IL-6 signal transduction pathway was also exhibited by EMSA. Results　Two IL-6 signal transduction pathways (JAK/STAT and Ras/NF-IL-6) can be activated by IL-6 of different dose in U266 cells. When one of the two signal transduction pathways was up-regulated, the other one was down-regulated. Conclusion　There is an antagonistic effect between the activation of two IL-6 signal transduction pathways in U266 cells.

Objective To investigate effects of driving fatigue on auditory involuntary attention.Method Between-groups design was used.The control group included 13 taxi drivers after adequate rest while the fatigue group included 13 taxi drivers who had been driving for 10 h.Auditory oddball pattern was adopted.The standard stimulus was 800 Hz,probability 80%;target stimulus was 1 000 Hz,probability 10%;novel stimulus was sound generated by computer or other sound,probability 10%.Subjects were asked to press the mouse upon hearing the target sound.Result The distribution of P3a was mainly around the frontal-central area of the subjects in control group;the amplitude of P3a was evidently lowered in subjects after driving fatigue.Conclusion The ability of auditory involuntary attention declines after driving fatigue.

Objective: To investigate which step of IL-6 signal transduction pathways in myeloma cells can be taken as the acting target of the antagonists for IL-6. Methods: EMSA and immunoprecipitation were used to detect the activation of transcription factors(TFs)-STAT3, NF-IL-6 and protein kinase ERK in a myeloma cell line-Sko-007 by IL-6. then anti-sense expression plasmids and anti-sense ODN for these signal moleculars were constructed and designed,the effects of these anti-sense nucleic acids on IL-6 signal transduction in Sko-007 cells were analyzed by the same methods. Results: IL-6 signal transduction could be antagonized by these anti-sense nucleic acids at different extent.Conclusion: TFs or protein kinases in IL-6 signal trareduction pathways can be taken as the target for antagonists design.

Objective:To compare the effects of repetitive transcranial magnetic stimulation (TMS) at three different relatively high frequencies on neuropathic pain so as to find the best frequency.Methods:One hundred Sprague-Dawley rats were randomly divided into a sham operation group, a model group, a TMS group and a control stimulation group. The TMS group was further divided into a 5Hz group, a 10Hz group, and a 20Hz group. The rats in the model, control stimulation and TMS groups received constriction injury of the sciatic nerve, while the rats in the sham group were given a sham version of the operation. On the third day after the operation the rats in the TMS group and the control stimulation group began to receive TMS treatment. Neuropatic pain was evaluated on the day before the operation, and on the 3rd, 5th, 7th, 10th and 12th days after the operation. The evaluations included the paw withdrawal thermal latency (PWTL) and the paw withdrawal mechanical threshold (PWMT).Results:The average PWTLs and PWMTs in all of the TMS groups increased with the TMS treatment. Those of the 10Hz and 20Hz groups were significantly higher than the 5Hz group′s average, while there were no significant differences between the 10Hz group and 20Hz group.Conclusions:High-frequency TMS at different frequencies has different effects on neuropathic pain, at least in rats. The treatment efficacy at 10 and 20Hz is superior to that at 5Hz.

Objective:To investigate the relationship between “activation of IL 6 signal transduction pathways” and “induction of CD45 expression” in a myeloma cell line—Sko 007 by IL 6.Methods:Electrophoretic mobility shift assay(EMSA) was used to detect the activation of the transcription facotrs STAT3 and NF IL 6 by IL 6,which can represent the activation state of the two IL 6 signal transduction pathways respectively;Then the induction of CD45 mRNA and protein expression by IL 6 were detected by RT PCR and FACS.The effect of antisense expression plasmid of STAT3 pAT STAT3 on STAT3 activation and CD45 expression was shown by EMSA and RT PCR,respectively.Results:①Both two IL 6 signal transduction pathways JAK/STAT and Ras/NF IL 6 can be activated by IL 6 in Sko 007 cells.②The expression level of CD45 mRNA and protein is Sko 007 cells can be up regulated by IL 6.③The activation of JAK/STAT signal transduction pathway and induction of CD45 mRNA expression by IL 6 can be inhibited in pAT STAT3 transfected Sko 007 cells.Conclusion:IL 6 induced CD45 expression in Sko 007 cells in mediated by the activation of JAK/STAT signal transduction pathway.

Objective:To investigate the mechanism of dexamethasone(DEX) induced apoptosis of human myeloma cell line Sko 007 and the inhibitory effect of IL 6 on apoptosis.Methods:Effect of DEX on the growth of Sko 007 cells was measured by MTT assay;Apoptosis of Sko 007 cells induced by DEX was determined by flow cytometry with propidium iodide(PI) staining of nuclei;The expression of three anti apoptotic Bcl 2 family proteins Bcl 2,Bcl X L and Mcl 1 in Sko 007 cells with or without DEX were determined by immunblot assay.Results:DEX inhibited the proliferation of Sko 007 cells and induced a significant cell apoptosis.In addition,degradation of Bcl 2 can be observed in Sko 007 cells in the presence of Dex.IL 6 stimulation prevented down regulation of Bcl 2 and the apoptosis of Sko 007 cells induced by DEX.Conclusion:IL 6 inhibits DEX induced apoptosis of Sko 007 cells through the specific regulation of Bcl 2.

An enzyme linked immunosorbent assay ( ELISA ) and a flow cytometry assay ( FCA ) based on Wil2-S cells were developed and systematically compared for quantification of recombinant anti-CD20 humanized monoclonal antibody ( rh-anti-CD20zumab) in biological matrix. The specificity, precision and accuracy of each method at correspondingly different linear range showed good results. For ELISA, the precisions of intra-day and inter-day were both <19 . 5%, the relative error was from-18 . 2% to 17 . 6%;For FCA, the precisions of intra-day and inter-day were both <19. 0%, the relative error was from -18. 9% to 18. 4%. The sensitivity of ELISA was significantly higher than that of FCA. The quantitative ranges of ELISA and FCA methods were 0. 04-5. 0 mg/L and 3. 1-200 mg/L, respectively. The concentrations in serum samples and pharmacokinetics analysis were determined by both of two methods after vein drip administration of rh-anti-CD20zumab in rhesus monkeys. Pharmacokinetics data showed that there was excellent consistency between results obtained by two methods at the given dose. We believe that the novel FCA with high speed and high sensitivity can be used to perform PK and PD study of cell surface antigen-targeted antibody derivatives.

Objective To investigate the effect of 36 h continuous sleep deprivation(SD) on circadian clock gene expression in the rat liver and kidney and the alteration of urine biomarker levels.Methods Twelve rats were randomly divided into control group and SD group.An SD device was used to deprive the rats of sleep.After 36 h continuous SD, the abdominal cavity was exposed to obtain livers and kidneys, and RT-PCR and Western blotting were used to detect expression of clock genes.Then,the pelvic cavity was exposed to obtain urine, and the changes in bio-marker total bile acids(TBA) were tested with ELISA.Results Compared with the control group, the mRNA level of liver clock and bmal1 was obviously reduced in the SD-treated rats (P<0.05).However, no obvious change was found in the samples from the kidney.Sharp down-regulation of CLOCK and BMAL1 protein expression was also observed in the rat liver after SD treatment.Urine TBA content in SD treated rats was raised obviously (P<0.001), compared with control.Conclusion Thirty-six hours of continuous SD could result in deregulation of circadian clock gene and cholesterol metabolism disorder in the rat liver.TBA might be used as a noninvasive biomarker of liver injury under SD stress conditions.

Objective To investigate the molecular mechanism of vascular endothelial growth factor ( VEGF) expression in bronchial epithelial cells (Beas-2B)induced by particulate matter 2.5(PM2.5).Methods PM2.5 powder was dis-solved in DMEM medium and diluted into five concentrations , 0,12.5,25,50 and 100μg/ml, respectively.The double an-tibiotics ( streptomycin and penicillin ) and FBS were added into the solution to a 2% final concentration of serum system after being treated by ultrasound for 30 minutes.The cultured Beas-2B cells were then treated with different doses of PM2.5.Subsequently, nuclear factor-kappa B(NF-κB) transactivity and the transcriptional activation of vegf gene promot-er were tested by dual-luciferase reporter gene analysis system while phosphorylation of p 65 , expression levels of IκBαand VEGF were detected by Western blotting .Results PM2.5 induced up-regulation of VEGF expression in Beas-2B cells in a dose-dependent manner , accompanied by NF-κB transactivation at the highest level under 100 μg/ml of PM2.5 treatment. Moreover, PM2.5 induced degradation of the repressor protein IκBαand increase in the phosphorylation level of p 65 sub-unit in Beas-2B cells.Knockdown of NF-κB p65 expression significantly inhibited vegf gene promoter transcriptional activa-tion as well as VEGF protein expression in Beas-2B cells induced by PM2.5.Conclusion PM2.5 induces VEGF expres-sion via activation of NF-κB pathway in bronchial epithelial cells .