Objective To explore a practicable method of serial sections of whole CNS in the rat, in which both neurons and nerve fibers were showed at the same section. Methods After perfused fixation and quick freezing, the tissues of whole CNS were cut into serial sections with cryostat followed by hematoxylin stain. Results The nerve fibers were stained in blue color while the neurons were showed black-blue and the background was light-yellow.Conclusion Our method was feasible to show both neurons and nerve fibers at the same section with quick freezing, freezing serial sectioning and improved hematoxylin staining.

Objective To investigate the effect of hypertension on carotid atherosclerosis ,and the clinical significance of BFI technique in the diagnosis of carotid atherosclerosis .Methods Using BFI technique and CDFI technique to detect 198 cases of pa-tients with hypertension(hypertension groups) and 200 cases with normal blood pressure(control group) of carotid internal-media thickness(IMT) and the number of atherosclerotic plaque .To explore the relationship between hypertension and carotid atheroscle-rosis .Results IMT and the detection rate of plaque of hypertension groups was significantly higher than which of control group (P<0 .01) .In hypertension groups ,the higher of blood pressure levels ,the higher carotid IMT and the detection rate of plaque(P<0 .01 or P<0 .05) .The detection rate of plaque in BFI technique was higher than which in CDFI technique (P<0 .01) .Conclusion hypertension can aggravate carotid atherosclerosis ,and the higher blood pressure levels ,the more obvious of carotid atherosclerosis . BFI technique is useful for the detection of carotid atherosclerosis .

Objective To explore the effect of hypoxia on the expression of Janus kinase (JAK) in cocultured pulmonary arterial smooth muscle cells (PASMC) Methods Rat PASMC were cocultured with lung microvascular endothelial cells (LMVEC), the expression of JAK protein in PASMC was detected with Immunocytochemical method Results Expressions of JAK1 and JAK3 Protein were seen excessive positive (++++) in the cytoplasm of simple PASMC in hypoxia, and better positive (+++) in cocultured PASMC in hypoxia Compared with those of the control group, the expressions of JAK1 and JAK3 protein obviously increased in PASMC in hypoxia for 2 h, reached highest in those with hypoxia for 6 h, and decreased in those with hypoxia for 12 hours hypoxia, but still higher than those in hypoxia for 2 h Conclusion Hypoxia modulates the signal pathways and improves cell proliferation by the enhancement of the expressions of JAK1 and JAK2 proteins in co cultured PASMC

To examine the effect of Gankang Suppository on duck hepatitis B virus (DHBV), the serum biochemistry and hepatic histology in an animal model of DHBV infection, a model of DHBV infection was established by infecting 1-day-old Yingtaogu ducklings with DHBV-positive serum. The successful model was confirmed by PCR assay and 48 ducklings infected with DHBV were randomly divided into 3 groups: a Gankang Suppository treatment group, an acyclovir (ACV) group and a DHBV model group (control), with each group having 16 animals. All the animals were given the medicines for 4 weeks in a row. The serum of the animals was taken 14 and 28 days after the medication and 7 days after drug discontinuation. Real-time PCR was performed to detect the copy numbers of DHBV DNA in the serum. ALT and AST were dynamically monitored. The ducklings were sacrificed on the 7th day after the discontinuation of the treatment and livers were harvested and examined for inflammation and degeneration of liver cells by using HE staining. The results showed that on day 14, 28 after the treatment and day 7 after the withdrawal, the logarithmic values (log) of DHBV DNA copy numbers in ducklings of Gankang Suppository treatment group were significantly lower than that before the treatment (P=0.0092, P=0.0070, P=0.0080, respectively). Compared with DHBV model control group, the ALT level was significantly decreased (P=0.0020, P=0.0019, respectively) on day 28 after the treatment and on day 7 after the withdrawal. The AST level was also reduced on day 14 after the treatment (P=0.0298). Compared with the ACV control group, the level of ALT was lower on day 7 after the withdrawal (P=0.0016). Histologically, the hepatocyte swelling, vacuolous degeneration and acidophilic degeneration in Gankang Suppository treatment group were alleviated 7 days after the withdrawal as compared with model control group (P=0.0282, P=0.0084, P=0.0195, respectively). It is concluded that Gankang Suppository can effectively suppress DHBV replication, reduce the levels of serum ALT and AST and improve hepatic histology.

A method was developed to elucidate the structures of sulfated oligosaccharides through establishment of an effective reversed phase ion pair ultra-performance liquid chromatography coupled with electrospray ionization time of flight mass spectrometry( RPIP-UPLC-ESI-TOF-MS). Heptylamine (20 mmol/L,pH4) has been selected as the ion-pairing agent.κ-Carrageenan oligosaccharides have been separated on BEH C_(18) column using MeOH/H_2O with 25% heptylammonium formate as eluent in linear gradient mode. Mass spectra were obtained by ESI-Q-TOF-MS in both positive and negative modes. κ-Carrageenan oligosaccharides were well separated up to pentatetrasaccharide,and ESIMS analysis for κ-carrageenan oligosaccharides up to hepta-cosasccharide. The results showed that all acid hydrolyzed κ-carrageenan oligosaccharides were odd sugars,which was further confirmed by polyacrylamide gel electrophoresis ( PAGE). The characteristic fragmentation pattern of ion-pair oligosaccharides in mass spectra can be applied for rapid structure identification.

BACKGROUND: NOV protein encoded by nephroblastoma overexpression gene(NOV) is IGF(insulin-like growth factor) -binding protein. What is its impact on human neural stem cell(hNSC) proliferation and differentiation?OBJECTIVE: To investigate the impacts of NOV protein on hNSCs proliferation and differentiation.DESIGN: A single factor analysis of variance experimental study using cells as subjectsSETTING: Department of histology and embryology, and department of neurobiology in a military medical university.MATERIALS: Study was conducted in the Department of Histology and Embryology of the Third Military Medical University of Chinese PLA. Subjects were hNSCs cultured from 10 to 14 weeks human embryo cerebral cortex.INTERVENTIONS: COS-7 cells were transfected by NOV gene recombined plasmid. COS-7 cell and COS-7 cell modified by NOV gene conditioned culture media(COS-CM and NOV-CM) were collected and reacted with the cultured HNSCs.MAIN OUTCOME MEASURES: hNSCs proliferation was detected by 3H-TdR scintillation analysis, and hNSCs differentiation was detected by immunocytochemistry and flow cytometer(FCM).RESULTS: Both COS-CM and NOV-CM could significant promote the intake of 3H-TdR by HNSCs, of which the 1/minute of NOV-CM group was significantly higher than that of COS-CM group(P ＜ 0.05), which indicated that NOV-CM contained component that could facilitate hNSCs proliferation, and moreover, there was certain dose-effect relationship in NOV-CM' s facilitation of cellular proliferation. The results of immunocytochemistry and FCM revealed that there were more NF-200 positive cells in NOV-CM group, while many glial fibrillary acidic protein positive cells could be seen in COS-CM group.CONCLUSION: NOV protein might have facilitative effects on hNSCs proliferation and differentiation into neurons.

Objective To explore the related risk factors of renal function after partial nephrectomy,and provide the reference for clin-ic. Methods The data of 31 patients with small renal cancer in our hospital from January 2010 to June 2014 were analyzed. Before and after the treatment,the relationship of renal glomerular filtration rate ( GFR) with the change and clinical pathological factors were analyzed,the single factor analysis used t test,multiple factors analysis adopted multi-factor unconditioned logistic regression analysis. Results Single fac-tor analysis results showed that renal function after partial nephrectomy was closely related to the age,diabetes mellitus,blocking time,the size of the tumor,renal volume reduction ratio,the difference was statistically significant (P<0. 05). Many factors of logistic regression showed that it was closely related to the age,blocking time,renal volume reduction ratio(P<0. 05). Conclusion The age,blocking time,renal vol-ume reduction ratio are risk factors for renal function after partial nephrectomy.

Objective To study the determination of biological activity of ?_1-AT with chromogenic substrate and its influential factors using fresh pooled normal human plasma as reference standard.Methods Measuring absorption value of reactions between residual trypsin and BAPNA at 405nm,after ?1-AT inhibition by excess trypsin.Fresh pooled normal human plasma was used as reference standard to calculate the biological activity of?1-antitrypsin.Results Good linear correlation was obtained when the plasma was diluted to 1/50 to 1/100.PEG4000,sucrose,S/D(Tween80/TNBP) and sodium caprylate did not influence the biological activity of ?1-AT,but?1-AT activity was increased by about 20% when the concentration of sodium citrate was above 0.125mol/L.Conclusion The experiment proved that?1-AT biological activity was determined using fresh pooled human plasma as reference standard,the method is stable and reliable.Except sodium citrate,all of the materials used in the assay did not influence the determination of?1-AT activity.

To study the effect of endotoxin on liver apoptosis, L02 liver cells were cultured and passaged in vitro, and then stimulated by endotoxin at 10 mg/mL for 4, 8, 16 and 24 h respectively. Liver apoptosis was flow cytometrically and fluorescently detected. Immunohistochemistry was used to detect the delivery of smac and caspase9. The delivery of liver cell smac and the activity of caspase3 were measured by caspase3 assay kit. The hepatic failure models of rats were established by using D-galactosamine. The blood serum and liver tissues were collected for the detection of the liver function, the level of endotoxin and the activity of caspase3 by using chromogenic substrate limulus amebocyte lysate method (LAL) and caspase3 active assay kit. The expression of smac and caspase9 in liver cells was detected by Western blotting. With in vitro study, the L02 cells stimulated by LPS condensed into conglobation and formed apoptotic bodies. After those cells were stained by hoechst, the apoptotic cells displayed blue color under the fluorescent microscope. The apoptosis rate was increased over time and the apoptosis was mainly of advanced stage. Meanwhile, the rate of smac delivery and activity of caspase9 and caspase3 were increased on L02 cell membrane. In vivo, hepatic failure and obvious endotoxemia were induced by injection of more than 200 mg/kg D-GalN. Hepatic mitochondria smac was reduced with dosage of D-GalN and, on the contrary, the activity of caspase3 was increased. D-GalN at 200 mg/kg increased Caspase9 while D-GalN at 300 mg/kg decreased caspase9. Mitochondria signal channel plays an important role in the endotoxin-induced apoptosis of hepatic cells by promoting the release of smac from mitochondria to cytoplasm and activating caspase9 and caspase3 in its low-level channel.
Apoptosis/*drug effects ; Carrier Proteins/*metabolism ; Caspase 3/metabolism ; Caspase 9/metabolism ; Cell Line ; Cells, Cultured ; Endotoxins/*pharmacology ; Liver/cytology ; Liver/*metabolism ; Liver/pathology ; Liver Failure/chemically induced ; Liver Failure/pathology ; Mitochondrial Proteins/*metabolism ; Rats, Wistar

Objective To study the effect of a new mucA gene mutation on the biofilm formation process and the morphology of matured biofilm of Pseudomonas aeruginosa. Methods The mucA gene of PAO1 was cloned into the Pseudomonas aeruginosa expression plasmid pUCP20. The recombinant plasmid was transformed to mucoid PA17 which contained a new mucA mutation. Positive clones of the plasmids were identified by enzyme digestion and sequencing. The expression levels of algD in the positive clones were assessed by semi-quantitative RT-PCR. The modified plate culture method was used to establish the biofilm models of PA17, PA17 with recombinant plasmid and PAO1 in vitro. Results Transformation was identified by the decreased expression of algD in positive clones. The rate of biofilm formation of the positive clones was between those of PAO1 and PA17. The irreversible adhesion occurred after 8 h and the matured biofilm was observed on day 6. The morphologies of PA17, PAO1 and PA17 with recombinant plasmid were the same. Conclusion The mucA gene mutation of PA17 delays the formation of irreversible adhesion of PA17 biofilm, but it has no effect on the morphology of matured biofilm.