Double immunohistochemistry staining method was used to visualize Leu-and Met-enkephalin-immunoreactive(L-ENK-ir and M-ENK-ir) terminals on oxytocin (OT)- and vasopressin(VP)-containing cells and their processes of the rat hypothalamus. The L-ENK-ir and M-ENK-ir nerve endings were found in various density in the supraoptic nucleus(SON), the paraventricular nucleus(PVN) and each of the accessory neurosecretory cell groups of the hypothalamus. The density of the L-ENK-ir endings paralleled to, but always higher than that of the M-ENK-ir terminals in the magnocellular neurosecretory cell groups. The density of the L-ENK-ir and M-ENK-ir endings was highest in the periventricular area(PeV), the anterior commissural nucleus(ACN), the dorsal medial and dorsal lateral accessory nuclei (DMN and DLN), the anterior and posterior fornical nuclei(AFN and PFN), moderate in the SON and PVN, and lowest in the nucleus circularis (NC) and the lateral hypothalamic perivascular nuclei(LHPVN). In all the cell groups, the L-ENK-ir and M-ENK-ir terminals which were found in contact with the OT- and VP-containing cells and dendrites were consistant with the density of the endings in the same area. These results indicate that ENK-ir afferents may be in extensive contact with the OT-and VP-containing cells and dendrites in the SON, PVN and the accessory magnocellular neurosecretory cell groups of the hypothalamus. These ENK-ir afferents probably play an important role in the regulation of the secretion of OT and VP.

The monoaminergic afferents to the magnocellular neurosecretory neurons in the hypothalamic supraoptic and paraventricular nuclei (SON and PVN) have been studied in many aspacts. In the present paper, double immunohistochemistry staining method was used to visualize catecholaminergic, i.e. tyrosine hydroxylase-immunoreactive (TH-ir), and serotonin- immunoreactive (5-HT-ir) terminals on oxytocin (OT)- and vasopressin (VP)- containing cells and dendrites not only in the SON and PVN, but also in most of the accessory magnocellular neurosecretory nuclei of the hypothalamus in the rat. Both the 5-HT-ir and TH-ir terminals were found in close proximity to the OT- and VP- containing cells and dendrites in various magnocellular neurosecretory cell groups, but differing in their density. There was fairly high density of the TH-ir endings in most of the accessory neurosecretory cell groups of the hypothalamus. Fairly high to moderate density of the 5-HT-ir terminals were mainly found in the accessory cell groups. It seems that the magnocellular neurosecretory cell groups are differently regulated by CA and 5-HT inputs.

PAP and ABC methods have been used to detect the Oxytocin-like (OXT-L) neurons in the hypothalamus of the rat and the results obtained were compared with the CT-HRP retrograde tracing study from the posterior pituitary to the hypothalamus by Ju et al. It was verified that the projections to the posterior pituitary came from the supraoptic nucleus, paraventricular nucleus, anterior and posterior fornical nucleus, perivascular cell groups, preoptic and anterior hypothalamic areas, the dorsal accessory cell groups, and the periventricular area, especially forming subependymal plexuses along the lateral wall of the third ventricle and beneath the interventricular foramen. By combining WGA-HRP retrograde tracing and immunohistochemistry methods, OXT-L neurons in all the above listed nuclei or cell groups were found to project to the posterior pituitary. The subependymal plexuses may serve as a link between the CSF in the ventricles and the posterior pituitary.

A sensitive ABC or PAP immunohistochemical technique visualized with glucose oxidase-DAB-nickel method was used to examine the distribution and characteristics of the supraependymal serotoninergic nerve fibers (5-HT-SEP) in the adult rat forebrain. Three types of fibers could be distinguished: a few thick fibers (about 0.9?m in diameter), large amount of intermediate fibers (about 0.4?m in diameter), and numerous thin fibers (about 0.1?m in diameter). Although the 5-HT-SEP were found in all areas of the forebrain ependyma, the density and distribution patterns of different fiber types varied. In addition, the correspondence between the densities of ependymal cilia and 5-HT-SEP indicates the modulation of the cilia activity by 5-HT-SEP.

Comparative investigation of the distribution of galanin-immunoreactive (GAL-ir) nerve fibers in the neural lobe of the human being, monkey, dog, cat and rat was studied immunohistochemically. The density of GAL-ir fibers in the posterior pituitary was highest in the dog and rat, moderate in the monkey and cat; and lowest in the human being. A dense accumulation of positive fibers appeared in the pituitary stalk bordoring the intermediate lobe. The fiber density in the central part of the posterior pituitary was slightly higher than at its periphery in the human and monkey. Moderate amount of fibers were found to spread all over the neural lobe of the dog and rat, with slightly higher density in the posterior part of the dog. In the cat, the highest density of GAL-ir fibers was found along the wall of the recess hypophysis. A slightly increasing fibers or fiber clumps were seen in the peripheral region of posterior pituitary in four species of the animals and human being. Some fibers were found surrounding the blood vessels or located in their walls.

Immunohistochemical method was used to investigate the morphology and distribution of the oxytocin immunoreactive (OXT-ir) neurons and their dendrites in the subependymal plexus (SEP) around the third ventricle (OXT-SEP). The OXT-ir neurons were of magnocellular neurosecretory type which often intercalated between the ependymal cells, their dendrites were generally long and straight and mostly paralleled to or traveled within the ependyma,some of them could protrude into the ventricle, and most of them appeared heavily vacuous and have a strong tendency of grouping into fascicles. The plexus was composed of dorsal and ventral parts, the former corresponding to the extent of the magnocellular paraventricular nuclei and was extensive and dense, the dendrites were well organized; the latter was more sparse. The cell bodies and dendrites within and between different parts might mix or contact with each other. The functional significance of the plexus is discussed.

HCV NS5B, acting as a RNA dependent replication enzyme,has emerged as an attractive protein used as a target for screenig of drugs against HCV NS5B, and plays an important role in HCV replication. In the report the gene expression of NS5B in E.coli was investigated. PCR was performed to gain the gene of HCV NS5B from plasmid pBRTM/HCV 1 which contains whole sequence of HCV, and the truncated NS5B gene containing no hydrophobic domain was cloned into pGEM Teasy vector. The gene of the truncated NS5B was cut from pGEM Teasy vector and cloned into E.coli expression plasmid pET 21b, then pET 21b NS5B was transfected into E.coli cells. The protein E.coli lysates were purified and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting assay. The RdRp activity of NS5B was examined by scintillation proximity assay (SPA). The truncated NS5B gene was successfully cloned into pET 21b. The results of SDS PAGE and Western blotting assay showed: ①the molecular weight of the expressed product was about 68 000 D, ②The truncated NS5B protein was existed in media of E.coli cells, ③The activity of NS5BDCT21 His from HCV 1b amounted to 6 900 cpm (total incorporation of approximately). These findings suggest that soluble NS5B can be successfully expressed in E.coli and could secret into media.

Leu-enkphalin-like(L-ENK-L)neurons of the hypothalamus and its adjacentarea of the rat were stained immunohistochemically.It has been found that besidesthe area previously reported in the literature L-ENK-L neurons are also present inSFO and the ependyma of the third ventricle.By combining WGA-HRP retrogradetracing and immunohistochemical method,L-ENK-L neurons in supraoptic nucleus,paraventricular nucleus,periventricular area and anterior fornical nucleus were foundto project to the posterior pituitary,in which some neurons squeeze in between theependymal cells.The subependymal neurons may serve as a link between the ven-tricle and the posterior pituitary.

Objective Discussion leukoreduction of red blood cells suspended in two different storage bag placement and he-molysis rate impact on the supernatant free hemoglobin (FHb),to ensure the clinical transfusion is safe and effective.Meth-ods Selected 20 donors to sample 400 ml whole blood per person to make leukodepleted red blood cells,which were evenly divided into 10 bags.The 10 bags were randomly divided into two groups,one to the upright position,one group of horizon-tal.The two groups were stored under the same conditions.Respectively,in the 7,14,21,28,35 day,randomly removed one storage bag from each group,FHb and red blood cell hemolysis rate were measured and analyzed statistically.Results FHb and hemolysis rate results stored in the first 21 days of testing,uprightgroup were (217.310±48.477)mg/L and (0.250± 0.056)%,respectively horizontal group (173.972±39.027)mg/L and (0.189±0.045)%,the results set upright than hori-zontal group,the results were statistically(t=3.114,P =0.003<0.05 and t=3.798,P =0.001<0.05),the difference was statistically significant.Conclusion In the blood storage period,storage bags can be placed horizontally to reduce the de-struction of red blood cells,blood storage is more favorable.

Objective: To compare the effects of pericardium patch aortoplasty and pulmonary patch aortoplasty for treating the infants with aorticcoarctation (AC) combining hypoplastic aortic arch in order to provide a better surgical choice in clinical practice.
Methods: A total of 57 patients with AC combining hypoplastic aortic arch treated in our hospital from 2009-01 to 2014-12 were retrospectively studied. The patients were divided into 2 groups: Pericardium patch aortoplasty group,n=26 and Pulmonary patch aortoplasty group,n=31. The changes of the pressure gradient at post-operation and follow-up period were compared.
Results: There were 2/57 (3.5%) patients died, 1 in Pericardium patch aortoplasty group by pulmonary hypertension crisis, the other 1 in Pulmonary patch aortoplasty group by respiratory distress syndrome. No renal failure or neurological complication occurred in neither groups. The cardiopulmonary bypass time, aortic clamping time, ventilator time and ICU stay time were similar between 2 groups,P>0.05. Selective cerebral perfusion time in Pericardium patch aortoplasty group was shorter than Pulmonary patch aortoplasty group (30.5 ± 8.6) s vs (35.6 ± 10.3) s,P<0.05. By ultrasound estimation, the post-operative AC pressure gradients were decreased than they were before, as in Pericardium patch aortoplasty group (9.5 ± 7.5) mmHg vs (39.9 ± 15.5) mmHg and in Pulmonary patch aortoplasty group (11.8 ± 11.3) mmHgvs (39.2 ± 14.5) mmHg, bothP<0.05; while post-operative pressure gradients were similar between 2 groups,P>0.05. Follow-up study was conducted in 51 patients for (17.6 ± 16.6) months, Pericardium patch aortoplasty group had 6 patients with re-stenosis, 3 of them would receive balloon angioplasty and 3 would be continuously followed-up; Pulmonary patch aortoplasty group had 6 patients with re-stenosis, 2 of them ifnished balloon angioplasty and their pressure gradients were obviously decreased, 4 would be continuously followed-up. Kaplan-Meier curves presented that Pulmonary patch aortoplasty group was superior to Pericardium patch aortoplasty group in re-stenosis occurrence during follow-up period.
Conclusion: Both pericardium patch aortoplasty and pulmonary patch aortoplasty were effective for treating the patients with AC combining hypoplastic aortic arch, the early post-operative efifcacy was similar, while the mid-term follow-up result was better in pulmonary patch aortoplasty.