The distribution of neuropeptide Yimmunoreactive (NPY-IR) fibers in the spleen of rat was investigated by using the immunohistochemical ABC method combine with glucose oxidase-DAB-nikel enhancement technique. The results demonstrated that the rat spleen is richly innervated by NPY-IR fibers. The NPY immunoreactive nerve fibers are mainly distributed along the branches of the arteries, especially surrounding the central arterioles. They are also observed in the connective tissue capsule, the lymphoid tissue of the white pulp, red pulp and marginal zone and around the splenic sinusoid. The positive fibers are closely related to the blood vessels and lymphocytes in the spleen, which suggested that these fibers may regulate the development and function of splenic lymphocytes. NPY may have a direct or an indirect effect on regulating blood circulation of the spleen and the lymphocytes.

In this study, the distribution of calcitonin gene-related peptide immunoreactive (CGRP-IR) nerve fibers in the spleen of the rat was investigated by using the immunohistochemical ABC method combined with glucose oxidase-DAB-nickel enhancement technique. The results showed that the rat spleen was richly innervated by CGRP-IR fibers. The nerve fibers containing CGRP were mainly distributed in the red pulp, especially surrounding the splenic sinusoid and small blood vessels. They were also observed in the lymphoid tissue of marginal zone, and occasionally in the periarterial lymphatic sheath (PALS). The positive fibers distributed in the spleen, which suggested that the fibers might modulate the development and function of splenic lymphocytes.

Guided by aim of improving the survival rate of the injuries, damage control surgery (DCS) have undergone developments from simple gauze packing to planned resuscitation and reoperation for nearly a century, which leads to a great improvement in the treatments. In recent years, the concept of DCS has been more and more widely accepted and applied in many fields other than abdominal trauma. Besides, the technologies have been constantly standardized and the procedures have been more scientific. At the same time, some remarkable progress has been made in the research of DCS, such as new concepts of damage control resuscitation, permissive hypotension, and new technologies such as resuscitative balloon aortic occlusion and hybrid emergency surgery system.

Objective To determine the best extraction process of ginkgo leaves with the total transfer rate of total flavonol glycosides and terpene lactones as the index.Methods The effects of ethanol concentration, solid-liquid ratio and extraction time on extraction process were investigated by orthogonal design method, and the contents of total flavonol glycosides and terpene lactones were detected by HPLC to calculate transfer rate.Results The optimum extraction conditions were as follows:85% ethanol refluxing and extracting for three times;the first time extracting with five-fold amount of solvent (V/W) for 3 hours;the last two times with three-fold solvent (V/W) for 2 hours.Conclusion This extraction process has the advantages of simplicity of operator, reason, energy conservation, high efficiency, and is suitable for industrial production.

In teaching of Microbiology, we introduced microbiological history and research experiences of scientists. Students had more interesting in learning and got some ideas of science research. Eighty percent of them got good points.

Objective:To investigate the mRNA expression of the WIF-1 gene and the methylation of its promoter in breast can-cer, and to determine the correlation between the epigenetic aberrant WIF-1 DNA methylation and the clinicopathological significance of WIF-1 in breast cancer . Methods:RT-PCR and sensitive methylation-specific-PCR (MSP) were used to detect WIF-1 mRNA ex-pression and the methylation of the WIF-1 promoter in 30 breast cancer samples as well as in tumor-adjacent tissue samples and 9 be-nign breast tissues. Results:The WIF-1 mRNA expression in 30 breast cancer samples significantly decreased compared with those of the other two groups. In addition, WIF-1 methylation was more frequent in breast-tumor tissues compared with those in tumor-free tis-sues. Meanwhile, WIF-1 mRNA expression in breast cancer tissues involved the abnormal methylation of its promoter. Clinicopatholog-ical correlation analysis showed that the abnormal methylation of the WIF-1 gene promoter was not associated with age, TNM stage, histotype, or lymph node metastasis. Conclusion:WIF-1 mRNA expression loss due to abnormal methylation may be a crucial factor in breast cancer development and can thus be used in the prognosis and progression of the disease.

Anelectrochemicallyreducedgrapheneoxide/goldnanoparticle-chitosan(ERGO/AuNP-CS) composite film modified glassy carbon electrode ( GCE) was constructed by directly electrochemical reduction of GO, and then assembly of AuNP-CS polycation on the surface. The surface morphologies of different modified electrodes including bare GCE, GCE/GO, GCE/ERGO and GCE/ERGO/AuNP-CS were characterized by scanning electron microscopy ( SEM ) . The differential pulse voltammetric behaviors of the electrodes were investigated, and the results indicated that the composite of ERGO/AuNP-CS exhibited excellent electrocatalytic oxidation activity to uric acid ( UA) molecule. In 0. 10 mol/L of phosphate buffer solution (pH=6. 5) with a scanning rate of 100 mV/s, the proposed composite film modified electrode showed a linear electrochemical response to UA in the range of 0 . 05-110 μmol/L with a detection limit of 12. 4 nmol/L ( S/N = 3 ). The electrode displayed good selectivity, reproducibility and stability in the determination of UA in human serum and urine samples with a recovery of 93 . 8%-104 . 1%. The detection results were agreed with those of conventional spectrophotometry and uricase Kit methods.

Objective To investigate the expression of hypoxia inducible factor-1α(HIF-1α)in the transplanted liver in rats and the role of HIF-1α in the JNK signaling pathway.Methods Ninety-six SD rats were randomly divided into normal control(NC)group,autotransplantation(AT)group,hypoxia preconditioning (HP)+ AT group according to simple random sampling method.No treatment was applied to the rats in the NC group except for blood vessel separation.Stable rat models of 70％ AT was established in the AT group.Rats in the HP + AT group were given 8％ oxygen mixed gas for 90 minutes before the operation.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected at 1,2,12,24 hours after operation,and the expression of HIF-1α in the hepatic tissue,mRNA expression of c-Jun,and protein expression of Cleaved Caspase-3 were detected by immunohistochemical staining,reverse transcription-polymerase chain reaction and Western blot,respectively.All data were analyzed using the analysis of variance or t test.Results The levels of ALT and AST in the HP + AT group were significantly lower than those in the AT group,while higher than those in the NC group at 1,2,12 and 24 hours after the operation(F=2631.371,1177.642,810.383,682.848;743.618,1095.522,375.995,580.613,P ＜0.05).The protein expression levels of HIF-1α in the AT group were significantly lower than those in the HP + AT group,but higher than those in the NC group at 1,2,12 and 24 hours after operation(F = 191.737,284.482,459.419,213.782,P ＜ 0.05).The mRNA expression levels of c-Jun in the HP + AT group were significantly lower than those in the AT group,while higher than those in the NC group at 1,2 and 12 hours after operation(F = 66.211,53.169,9.645,P ＜ 0.05).There was no significant difference in the mRNA expression of c-Jun among the 3 groups at 24 hours after operation(F = 1.100,P ＞ 0.05).The protein expressions of Cleaved Caspase-3 in the HP + AT group were significantly lower than those in the AT group,while higher than those in the NC group at 1,2,12 and 24 hours after the operation(F =23.133,31.158,14.347,29.043,P ＜ 0.05).Conclusion High expression of HIF-1α after HP inhibits apoptosis of hepatic cells and alleviates ischemia reperfusion injury of hepatic tissues by suppressing JNK activation,down-regulating protein expression of Cleaved Caspase-3 after orthotopic liver transplantation in rats.

Objective To investigate the effect of hypoxic preconditioning on hepatocytes early growth response factor 1 ( EGR-1 ) in a rat liver autotransplantation. Methods The rat portal vein perfusion model was established for donor liver autotransplantation. Rats were then divided into group A:hypoxic preconditioning was done before transplantation; group B: rats undergoing liver transplantation without preconditioning; group C: Normal control group of rats. Liver histopathological changes, the mRNA expression of HIF-1, TNF-1 and the WB results of EGR-1 were compared between groups. Results The expression of HIF-1 α RNA determined in group A was more obvious than in group B and group C. Six hours after surgery, the expression in group A was significantly higher than that in group B (t =9. 601, df= 10, 2-tailed Sig = 0. 000, P<0. 05 ) ; Egr-1 protein expression in group A and group B increased after surgery,with that in group A being significantly lower than that in group B. The RT-PCR expression of TNF-α RNA in group B compared with group A and group C was more obvious. Six hours after operation, the expression of TNF in group B was significantly higher than that in group A ( t = -12. 067, df = 10, 2-tailed Sig =0. 000, P<0. 05 ). The expression of Egr-1 was positively correlated with that of TNF. A liver cell pathology showed less severe injury in structure of hepatic lobule, mild swelling of liver cells, no significant changes in liver tissue. Conclusions Hypoxic preconditioning adaptation in rat liver transplantation generates modest increase in EGR-1, and reduces the production of TNF and other inflammatory factors.

Objective To investigate the effect of nerve growth factor (NGF) on osteogenesis induced by bone morphogenetic protein-9 (BMP-9) in mouse embryonic fibroblasts (MEFs).Methods MEFs were respectively transfected with adenovirus-mediated NGF (NGF group), BMP-9 (BMP-9 group) and NGF + BMP-9 (combined group) and green fluorescence protein (GFP) (control group).Cytochemical staining was used to test the activity of alkaline phosphatase (ALP) 3 d and 5 d after treatment.Level of osteopontin (OPN) mRNA was detected by RT-PCR 9 d after treatment.Level of OPN protein was assayed by Western blot and immunocytochemistry 9 d after treatment.Mineralization was detected by Alizarin red staining 14 d after treatment.Results ALP activity in MEFs was elevated in BMP-9 group rather than in NGF group, but a significant increase in ALP activity was noted in combined group.In control group, BMP-9 group, NGF group and combined group, level of OPN mRNA was 0.92 ± 0.03, 1.28 ± 0.04, 0.94 ± 0.03 and 1.62 ± 0.04 respectively (F =214.60, P ＜ 0.01);level of OPN protein was 0.60 ± 0.05, 0.84 ± 0.03, 0.53 ± 0.05 and 1.27 ± 0.05 respectively (F =162.5, P ＜ 0.01).In comparison, OPN mRNA and protein were significantly up-regulated in combined group than in BMP-9 group (t =10.569 and 11.778,P ＜ 0.05).In control group, BMP-9 group, NGF group and combined group, relative density of OPN protein was 3.63 ±0.17, 6.27 ±0.30, 3.86 ±0.18 and 10.16 ±0.18respectively (F =602.6, P ＜ 0.01), with a significant higher level in combined group than in BMP-9 group (t =22.280, P ＜ 0.05).Level of mineralization was significantly higher in combined group than in BMP-9 or NGF group.Conclusion NGF can potentiate the osteogenesis induced by BMP-9 in MEFs.