Immunohistochemical method, immuno-electron microscopic technique and radioimmunoassay (RIA) were used for the demonstration of ?-endorphin-like immunoreactivity (?-EP-IR) and of the circadian variation of the ?-EP-IR level in rat pituitary. ?-ER-IR was located in all cells of the pituitary intermediate lobe. A few ?-ER-IR-positive cells were scattered in the anterior lobe, and no positive immunoreactivity was found in the posterior lobe. The controls were negative. Under electron microscope, ?-ER-IR-positive dense particles were mostly located in the secretion granules, and a few of them were scattered freely in the cytoplasm of the intermediate lobe cells. Both pituitary ?-EP-IR concentration and the basal pain threshold in rat showed circadian variations in the experiment done at 4-hr intervals over a 24-hr period (P

BACKGROUND: Mutation of the mitochondrial DNA may occur during the aging process of organisms, which is especially likely in the central nervous system. Evidences have been obtained that mitochondrial dysfunction may ensue from genetic impairment involved in oxidative phosphorylation, which is accompanied by corresponding morphological changes.OBJECTIVE: To investigate the association between olfactory disturbance and ultrastructural mitochondrial changes in olfactory bulb neurons of aged rats in comparison with young rats.DESIGN: Randomized controlled experiment.SETTING: Department of Internal Medicine, Affiliated Hospital of Chende Medical College and Department of Electron Microscopy of Chende Medical College.MATERIALS: This experiment was conducted in the Department of Electron Microscopy of Chende Medical College between April and December 2002. Sixteen male Wistar rats were divided equally into aged group (＞ 24 months) with body mass of 300-350 g and young group (6 months) with body mass of 180-220 g.METHODS: The rats in the two groups were anaesthetized by intraperitoneal injection of 10 g/L urethane (1 g/kg) and the chest was opened to insert a tube into the ascending aorta for perfusion with 200 mL of the mixture containing glutaric dialdehyde and paraformaldehyde for fixation.The olfactory bulb was then obtained and sliced, fixed in perosmic acid and embedded. Each layer of the olfactory bulb was observed under optical microscope and ultra-thin sections were prepared for observation under transmission electron microscope.MAIN OUTCOME MEASURES: The stratification of rat olfactory bulb and ultrastructural changes of the mitochondria in the major neurons in the olfactory bulb.REULSTS: No obvious changes were found in the stratification of the olfactory bulb in the two groups. From the exterior to the interior of the olfactory bulb, the olfactory nerve fiber layer, glomerular layer, external plexiform layer, mitral cell lalyer, internal plexiform layer and olfactory bundle nerve fiber layer were observed. Ultrastructurally, the mitochondria were more numerous in the brush cells in aged rat olfactory bulb, appearing rodshaped or ovoid with irregular cristae which were arranged in local disorder and some fragmented into small granules to form small bodies of high electron density. A few mitochondria became myeliuoid body with irregular or concentric circular morphologies. In the younger rats, most of the mitochondria in the neurons were ovoid with distinct outer and inner membrane and regular arrangement of the mitochondrial cristae, and irregular arrangement of the cristae was seen only in rare cases.CONCLUSION: The stratification of the olfactory bulb is basically the same in aged and young rats, but the ultrastructural changes in the mitochondria in the major neurons in aged rat olfactory bulb can be obvious,which is consistent with the reduced mitochondrial oxidative phosphorylation function. This might be one of the major causes for olfactory disturbance related to aging.

Objective　The microvasculature of the anterior lobe, posterior lobe and cerebellar vermis was observed under the optical microscope in 6 rats. Methods　The tannic acid-ferric chloride method (TAFM) was used to stain the vessels in cerebellum. Results　The arteries within the cortex and medulla of the cerebellum originated from pia mater arteries and central branches of the cerebellar arteries, the branches of pia mater arteries mostly penetrated into the cerebellar cortex vertically and formed capillary net in the molecular layer, purkinje layer and granular layer, the arteriole in the medulla of cerebellar posterior lobe branched and formed clawed-like figurations. These vessels beneath lobule cortex connected mutually by vessel anatomosis across the medulla. Conclusion　The arteriole in the cerebellum can be manifested distinctly in a strong three-dimension by TAFM and the vessel anatomosis in deep layer of cerebellum can be easily observed.

Objective To observe the inhibitory effect of xiaoaiping injection and octreotide on H22 tumor-bearing mice and find the best drug concentration,then to explore its mechanism.Methods Establish a mouse H22 subcutaneous tumor model.After tumor the experiment animals were divided into normal control group,model group,Xiaoaiping low,medium and high dose group,octreotide group,and the group of XAP low,medium and high dose groups were combined with OCT.Calculate the tumor's volume and draw the tumor growth curve.Intraperitoneal injection for 14 days,Inhibitory rate was calculated; To observe its pathological changes by light microscope; The ratio.of CD3 + NK1.1-T cells,CD3-NK1.1 + NKcells,CD3 + NK1.1 + NK-Tcells in peripheral blood were measured by flow cytometry.Results Compared with the control group,H22 liver cancer in different treatment group had a certain inhibition effect on growth,The inhibitory effect of the combination group was better than single-agent group,High-dose Xiaoaiping + octreotide was best,Tumor model group compared with normal control group,The ratio of T cells,NK cells and NKT cells was significantly lower(P ＜0.05) ; T cells,NK cells and NKT cells after treatment in each group had some enhancement,High-dose Xiaoaiping + octreotide was the most obvious,the ratio of T cells,NK cells and NKT cells of the combination group was significantly more than the single-drug group and the same concentration of octreotide monotherapy Xiaoaiping group(P ＜ 0.05).Conclusion High-dose Xiaoaiping + octreotide is the best drug for the inhibitory drug concentration.The inhibition of tumor growth may pass to improve the tumorbearing mice with immune status and enhance the body's anti-tumor capacity.

To enhance interns' comprehensive ability and qualification in medical laboratory , ISO 15189 quality system that was widely accepted at home and abroad was applied in practice teaching management. First teaching experience was shared with each other, teaching competition was encouraged to improve the qualification of teaching, and then relevant academic conference and the application of scientific or new projects were also advocated to promote the construction of teaching staff. Second, the management system of the intern was improved, the rules of the management of the intern files were optimized, the train-ing and study were regularly organized, the teaching system was perfected, and the practice appraisal was standardized. Practice proved that the graduates had strong ability to work, to master the quality management of ISO 15189 elements and could solve the common problems in clinical work and at the same time they had certain scientific research thinking.

Objective:To analyze the impact of NBP on the expression of HSP 70 and TLR4 in the rat with ischemia-reperfusion.Methods:The 144 rats were divided into 3 groups,Sham group,IR group and NBP group.The 3 groups were divided into 4 groups again according to the time of reperfusion (6h,12h,24h and 48h).The intraluminal ischemia-reperfusion model was made by improved Longa.When the model was made successfully ,the rat in NBP group was gave oral NBP immediately while the IR and Sham group was gave oral NS.The neurological deficit scores , number of apoptotic cells , expression of HSP70 and TLR4 were compared during the 3 groups.Results:The neurological function ,apoptosis,the expression of HSP70 and TLR4 in NBP group at any reperfusion time were statistical lower than in IR group.In all the infusion periods,apoptotic cells in IR and NBP group were significantly higher than that in the Sham group.In all the infusion periods ,the positive cells in IR and NBP group were significantly lower than that in the Sham group.Conclusion:NBP can protect the function of neuron by improving the inhibition of HSP 70 and TLR4.

Objective To investigate the expression of adenosine receptor (ADOR) subtypes (A2A and A2B subtypes) in the mucosal dendritic cells (DCs) from patients with Crohn's disease and their pathogenic roles.Methods Mucosal DCs (mDCs) were isolated from resected intestine of patients with or without Crohn's disease.Some of the mDCs were cultured in vitro and others were used to extract RNA.The expression of ador-a2a and ador-a2b were detected by real-time qPCR.mDCs in culture were treated with selective ADOR-A2A and ADOR-A2B agonists (CGS 21680 and BAY 60-6583) and then the concentration of IL-1,IL-6 and IL-12 in the medium were measured by ELISA.The binding affinities of ADOR-A2A and ADOR-A2B to adenosine were determined by 3H-adenosine in combination with selective ADOR-A2A and A2B antagonists (SCH58261 and MRS1706).Na(i)ve CD4+ cells were collected from human umbilical cord blood and co-cultured with mDCs treated by different ADOR agonists to observe T cell responses.The production of cytokines in culture was measured by ELISA.The polarization of CD4+ cells was analyzed by intracellular cytokine staining and FACs analysis.Peripheral blood mononuclear cells (PBMCs) were treated with IL-4 and GMCSF to induce the expression of monocyte-derived DCs (Mo-DCs).Mo-DCs were treated with different toll-like receptor ligands to investigate their effects on the expression of ador-a2a and adora2b.Moreover,Mo-DCs were treated with LPS and BAY 60-6583 individually or in the combination to stimulate CD4+ cells.Then the production of cytokines and the polarization of CD4+ cells were evaluated.Results Compared with patients without Crohn's disease,patients with Crohn's disease showed no change in the expression of ador-a2a but a significantly increased expression of ador-a2b in mCDs (CD-mDCs),enabling to bind more adenosines.Activated ADOR-A2B signaling pathway induced CD-mDCs to secret more proinflammatory cytokines and to promote polarization of CD4+ cells toward Th1 and Th17 cells.Toll-like receptor ligands,pam3csk4 and LPS could intensively augment the expression of ador-a2b in Mo-DCs.The pathogenicity of Mo-DCs was strengthened upon a combined stimulation with BAY 60-6583 and LPS.Conclusion The significantly increased expression of ador-a2b in mDCs might be involved in the pathogenesis of Crohn's disease by promoting mDCs to secret more pro-inflammatory cytokines and enhancing the polarization of CD4+ cells.Moreover,the expression of ador-a2b in DCs could be regulated by certain toll like receptors.

Objective To investigate the role of S100 calcium binding protein A12 (S100A12) in the pathogenesis of preeclampsia. Methods Sixty patients with preeclampsia were recruited from March 2013 to December 2013 in the First Affiliated Hospital of Zhengzhou University. Among them, thirty cases were defined as the mild preeclampsia group and thirty cases were defined as the severe preeclampsia group. The other thirty healthy pregnant women were recruited in the healthy pregnant women group. The levels of S100A12 protein in maternal peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry of streptavidin peroxidase biotin (SP) method was used to measure the protein expression of S100A12. The trophoblast cells were cultured in vitro with plasma from the three groups, and a blank control group was set up as well. Transwell was used to detect the cytotrophoblast invasion ability. Western blot was used to measure the protein expression level of receptor for advanced glycation end products (RAGE). Results (1) The levels of S100A12 in maternal peripheral blood of patients with preeclampisa [mild group:(30.8 ± 2.7)μg/L, severe group:(49.3 ± 4.1)μg/L] were significantly higher than that of the control group [(15.8 ± 1.4) μg/L]. In addition, compared with the mild preeclampsia group, the level of S100A12 in the severe preeclampsia group was significantly higher (P<0.05). (2)Positive immunostaining of S100A12 was observed in the cytoplasm of cytotrophoblast, decidual cells and the placentas from the three groups. The positive rate in the mild preeclampisa group was 77%(23/30);in the severe preeclampsia group it was 93%(28/30);and in the healthy pregnant women group it was 23%(7/30). The positive rates of placenta in the mild and severe preeclampsia groups were significantly higher than that in the healthy pregnant women group (P<0.05). In addition, compared with the mild preeclampsia group, the positive rate of immunostaining of S100A12 in the severe group was significantly higher (P<0.05).(3) Cytotrophoblast invasion ability and the expression of RAGE in the mild preeclampsia group were 29.1±3.2 and 0.479 ± 0.038, respectively;in the severe preeclampsia group they were 16.8 ± 2.5 and 0.652 ± 0.059;in the healthy pregnant women group they were 38.6 ± 24.3 and 0.327 ± 0.024; and in the blank control group they were 42.6 ± 5.6 and 0.194 ± 0.011. Cytotrophoblast invasion ability and the expression of RAGE protein in the mild and severe preeclampsia groups were significantly higher than those in the healthy pregnant women group and the control group(P<0.05). Conclusions The expression of S100A12 increased in materal peripheral blood and placenta, and the receptor protein of S100A12 RAGE also had high expression. It suggested that the S100A12 may have some effect on the pathogenesis of preeclampsia.

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link betw een sample donors and actual criminal acts. How ev-er, the conventional body fluid identification methods are prone to various limitations, such as time con-sumption, intensive labor, nonparallel manner, varying degrees of sensitivity and limited specificity. Re-cently, the analysis of cell-specific messenger RNA expression (mRNA profiling) has been proposed to supplant conventional methods for body fluid identification. Since 2011, the collaborative exercises have been organized by the European DNA Profiling Group (EDNAP ) in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification. The major advantages of mRNA profil-ing, compared to the conventional methods, include higher sensitivity, greater specificity, the ability of detecting several body fluids in one multiplex reaction, and compatibilitywith current DNA extraction and analysis procedure. In the current review ,we provided an overview of the present know ledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possi-ble practical application to forensic casew ork.

Objective To investigate Insertion/Deletion (InDel) polymorphism on the X chromosome and to screen 18 InDel loci for the Chinese Han population as a forensic DNA typing system auxiliary. Meth-ods Eighteen X-InDel markers were selected using the Human Genome Browser and dbSNP database. Multiplex PCR primer pairs of selected X-InDel markers were designed using Primer 3 software and di-vided into 3 groups according to the amplified fragment length, labeled by FAM, HEX and TAMRA fluorescence dye, respectively. The population genetics research and comparative analysis of Chinese Han nationality and 4 main minorities, the Hui, Wei, Mongol, and Tibetan nationalities, were investigated with the system. Results A new multiplex genotyping system, named InDel X-18PLEX, was successfully developed and validated, consisted of 18 X-InDel markers on the X chromosome and 1 Amelogenin gen-der marker. No deviation from Hardy-Weinberg equilibrium expectations was detected in the distribution of genotypes in the 5 investigated ethnic groups. However, there was significant difference between their distributions. From the investigation of Han nationality, high female (0.999 999 4) and male (0.999 88) overall discrimination power values were obtained, as well as high overall mean exclusion chance values in trios (0.999 992) and in duos (0.99). Conclusion InDel X-18PLEX meets the requirements as a forensic DNA complementary kit, providing effective supplementary analytical tools for difficult cases.