Objective To explore the preventive effects of Radix et Rhizoma Rhei Granule on the occurrence of complicated stress ulcer of stomach (CSUS) in rats after cerebral infarction (CI) and its mechanism. Methods Rat models wi t h CI were established by the occlusion of middle cerebral artery. Preventive tre at ment of CSUS in rats after CI with Radix et Rhizoma Rhei Granule was carried out . Results As compared with the control group (Group A),apoptosis index of gastric mucosal cells was decreased and prolifer ation index increased, 5_HT level and noradrenaline (NE) content in gastric muco sa and plasma were declined, gastric and cholic acid reduced and pH value incre a sed in gastric juice of the treatment group(Group B) (P

Objective To study the effect of Yiqi Huoxue Tablets on blood coagulation and fibrinolysis system of rats with cerebral apoplexy induced by artificial cold wave. Methods SD rats were randomized into 4 groups. The apoplexy-prone renovascular hypertensive rat model was established by the bilateral renal arteries clamping method. From the 14th day after the modeling operation, the treatment group was given Yiqi Huoxue Tablets (1.89 g·kg-1·d-1), the positive control group was given aspirin (80 mg·kg-1·d-1), and the model group and the blank group were given the same volume of distilled water, once a day. After 112 days, the rats were put into the artificial cold wave box staying for 3 days to inducing apoplexy, and then thrombin-activatable fibrinolysis inhibitor (TAFI) and protein C ( PC) , thrombin-antithrombin complexes ( TAT) and plasmin-antiplasmin ( PAP) levels were measured. Results The plasma levels of TAFI, PC, TAT, and PAP in Yiqi Huoxue Tablets group were lower than the model group, the difference being statistically significant ( P<0.05) . Conclusion The preventive and therapeutic mechanism of Yiqi Huoxue Tablets for cerebral apoplexy is probably related with the regulation of the balance between blood coagulation and fibrinolytic system of apoplexy-prone renovascular hypertensive rats.

Purpose　To observe the changes of dynorphin-like immunoreactivities of neurons in some rat brain nuclei that are related to analgesia following exogenous administration of melatonin.　Methods　The experimental rats were divided into two groups, injected intraperitoneally with melatonin 110 mg/kg and with vehicle, respectively. One hour after the injection, the rat brain was processed for coronal sections. The sections were stained with immunohistochemical ABC technique. The integral optical density (IOD) of the stained section was measured by the computer-assisted image processing technique.　Results　Dynorphin-like immunoreactivities in the supraoptic nucleus and nucleus raphe dorsalis showed obvious reduction following the single injection of melatonin.IOD values in the above nuclei were decreased significantly (P<0.01) with the melatonin treatment. In the paraventricular nucleus of hypothalamus, periaqueductal gray and nucleus raphe magnus, there was no difference (P>0.05) about the IOD values between melatonin-treated group and vehicle-treated group.　Conclusions　Melatonin may result in the decrease of dynorphin content in the supraoptic nucleus and nucleus raphe dorsalis.

[Objective] To observe the apoptosis of cortical cells and expression of related gene bcl-2 in hippocampal gyrus and frontal lobe of rats with incomplete cerebral ischemia (ICI) and to investigate the effects of Yiqi Huoxue Tablet (YHT, mainly composed of Radix Astragali , Rhizoma Chuanxiong, Radix Angelicae Sinensis, Herba Leonuri, Rhizoma Acori Tatarinowii, Scorpio, etc) . [ Methods ] Rats were randomized into six groups: model group, mimic operation group, nimodipine group and low-dose, moderate-dose and high-dose YHT groups. Flow cytometry was used to detect the apoptotic rate and bcl-2 protein content in cortical cells. Histological changes were also observed under light microscope. [Results] Early apoptotic changes were found in the model group and the apoptotic rate and bcl-2 protein content were higher than those in the mimic operation group; after treatment of YHT, the apoptotic rate in frontal lobe was decreased as compared with that in the model group and that in hippocampal gyrus remained higher than that in the mimic operation group and bcl-2 protein content was reduced to the level of the mimic operation group. [ Conclusion ] ICI is related to the apoptosis of nerve cells and YHT has been proved an effectual drug for the apoptosis.

During the production of ε-poly-L-lysine (ε-PL) in fed-batch fermentation, the decline of ε-PL synthesis often occurs at middle or late phase of the fermentation. To solve the problem, we adopted two strategies, namely pH shift and feeding yeast extract, to improve the productivity of ε-PL. ε-PL productivity in fermentation by pH shift and feeding yeast extract achieved 4.62 g/(L x d) and 5.16 g/(L x d), which were increased by 27.3% and 42.2% compared with the control ε-PL fed-batch fermentation, respectively. Meanwhile, ε-PL production enhanced 36.95 g/L and 41.32 g/L in 192 h with these two strategies, increased by 27.4% and 42.48% compared to the control, respectively. ε-PL production could be improved at middle or late phase of fed-batch fermentation by pH shift or feeding yeast extract.
Batch Cell Culture Techniques ; Fermentation ; Industrial Microbiology ; Nitrogen ; chemistry ; Polylysine ; biosynthesis

Objective To evaluate the role of recombinant human soluble Tim-3 (hTim-3-Fc) in regulating immune response.Methods Soluble hTim-3 was incubated with human macrophage cell line U 937, human T cell line Jurkat and normal human PBMC before cytokines secreted by or expressed in different immune cells were analyzed using ELISA , RT-PCR and Western-blotting, respectively.Results Soluble hTim-3 significantly promoted the activation of different immune cells.Our data showed that IL-8 secretion by U937 cells, IL-2 secretion by Jurkat cells , IL-2 and IFN-γsecretion by human PBMCs were all significantly increased .In addition , soluble hTim-3 significantly increased the IFN-α2 and IFN-β1 mRNA expression in U937, Jurkat and PBMCs and increased the phosphorylation of stat-1 in Jurkat and U937 cells.Conclusion Recombinant soluble hTim-3 can significantly promote the activation of immune cells in vitro, which shows its therapeutic potential .

Objective To examine whether Tim-3 plays a protective role in paraquat poisoning induced excessive immune response and tissue damage based on the critical roles of Tim-3 controlling inflammatory response.Methods A paraquat poisoning model was established in wild type and in Tim-3 transgenic C57BL/6 mice by intraperitoneal injection of paraquat (40 mg/kg) .In addition, C57BL/6 mice with paraquat poisoning were injected with Tim-3 soluble protein( sTim-3) or control protein to see the effect of Tim-3 blocking on the progression of paraquat poisoning.Samples were collected at 6 and 24 h after paraquat injection respectively and were examined for tissue damage, cytokine expression and paraquat metabolism.Results After paraquat poisoning, there was significantly attenuated tissue damage in the lungs and kidneys and decreased TNF-α,IL-6 and IL-1 beta expression in the PBMCs or in the serum from Tim-3 transgenic mice compared to wild type mice.The serum concentration of paraquat in Tim-3 transgenic mice was also significantly decreased.However, in sTim-3 treated paraquat poisoning mice, there was significantly increased cytokine expression and tissue damage compared to control protein treated mice.The in vitro data showed that Tim-3 signaling negatively regulated macrophages mediated inflammatory response.Conclusion Tim-3 plays a critical role in maintaining the homeostasis after paraquat poisoning. Further investigation on the regulatory roles of Tim-3 in inflammation will shed new light on the pathogenesis of paraquat poisoning and provide new therapeutic strategies.

Objective To develop a human Tim-3 specific monoclonal antibody and evaluate its biological activity and possible use in clinical diseases associated with dysregulated Tim-3 expression .Methods The BALB/c mice were immu-nized by conventional method, and positive clones were used to develop anti-human Tim-3 antibody, the binding and neutralization activities of which in vitro and in vivo were investigated.Results ①A monoclonal antibody (clone L3D) which could specifically bind to human Tim-3 protein in ELISA assay was obtained and the subtype of the monoclonal antibody was IgG2a .②Flow cytometry indicated that the monoclonal antibody could bind to Tim-3 expressed in human U937 cells.This antibody also showed a cross activity to mice′Tim-3.③The monoclonal antibody inhibited the apoptosis of THP1 cells induced by Gal-9, the ligand of Tim-3.④Injection of Tim-3 antibody exacerbated sepsis in mice as marked by the decreased survival rate and increased expression of pro-inflammatory cytokines .Conclusion An anti-human Tim-3 monoclonal antibody is successfully obtained.The excellent binding and neutralization activities of this antibody enable it to be widely used in clinical diseases associated with deregulated Tim-3 expression .