Objective To observe the changes of intracellular free calcium and the expression of CaM in the hippocampal neurons of posttraumatic stress disorder (PTSD) rats and to further investigate the neurobiological mechanisms. Methods The SPS-method was used to set up the rat PTSD models. A total of sixty male Wistar rats were randomly divided into 12 hours,1 day,4 days,7 days groups of SPS and normal control group. The intracellular free calcium was examined by fluorescence spectrophotometer. The expression of CaM was detected by using immunohistochemistry, Western blotting and RT-PCR. Results The intracellular free calcium level in the hippocampus of experimental rats was markedly increased 12 hours after SPS stimulation,and reached the peak after 1 day, then gradually decreased to normal level after 7 days. The expression of CaM in the hippocampus 1day after SPS was also the highest and then gradually decreased.Conclusion The lasting dysfunction of Ca~(2+)/CaM signaling cascades in hippocampal may play important roles in the pathogenesis of PTSD rats.

Objective:To investigate the protective effect of N-acetylcysteine(NAC)on right ventricular remodeling in a rat model of monocrotaline-induced pulmonary arterial hypertension ( PAH ) .Methods: PAH rats were induced by a single injection of monocrotaline(60 mg/kg,sc)and were ad ministered with NAC[500 mg/(kg? d)]for 6 weeks.At the end of 4 weeks,the right ventricular systolic pressure ( RVSP ) and mean pulmonary artery pressure ( mPAP ) were monitored via the right jugular vein catheterization into the right ventricle .Right ventricle ( RV ) to left ventricle ( LV )+septum ( S ) were calculated.Right ventricular morphological change was observed by HE staining .Sirius red was used to demonstrate collagen deposition .The expressions of collagenⅠ,collagen Ⅲ, NADPH oxidase 4 ( NOX4 ) and nuclear factor-kappa B ( NF-κB ) were analyzed by RT-PCR and ( or ) Western blot.Results:NAC attenuated RVSP ,mPAP and right ventricular remodeling index ( RV/LV+S) of PAH rats induced by monocrotaline after treatment for 4 weeks.Furthermore ,monocrotaline-induced right ventricular collagen accumulation and collagen Ⅰand collagenⅢexpression were both significantly suppressed by NAC .The expressions of NOX 4 and NF-κB were obviously decreased in right ventricule from PAH rats with NAC treatment.Conclusion:NAC ameliorates right ventricular remodeling of PAH induced by monocrotaline in rats through down regulating the expression of NOX 4 and antioxidant activity ,and inhibiting activation of NF-κB and collagen accumulation .

OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.

RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).

CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.

Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism

A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor (GDNF) on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons (SMNs) were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyltransferase (ChAT) expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group (P<0.05). This protocol can be adapted for various postnatal motor neurons studies.

To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.

ObjectiveTo identify the factors influencing the transfer of porcine endogenous retroviruses across the membrane of a new bioartificial liver (BAL),which is a necessary caution to consider for BALs carrying porcine hepatocytes.MethodsA novel porcine BAL composed of two circuits was constructed using a plasma filter with membrane pore size of 500 nm and a plasma component separator with membrane pore sizes 10 nm,20 nm,30 nm,and 35 nm.Co-cultured cells of porcine hepatocytes and mesenchymal stem cells or single porcine hepatocytes were incubated in the bioreactors.The BAL was continuously worked for 72 hours during which the supernatant was collected from the internal and external circuits every 12 hours.PERV RNA,reverse transcriptase (RT) activity,and in vitro infectivity from the supernatant were detected.ResultsIn the plasma filter group,the PERV RNAlevels were the same in both circuits,suggesting little to no effect of the plasma filter on the PERV RNA's crossing.With plasma component separators,PERV RNA was found in the external circuits at different times without positive RT activity and HEK293 cell infection,but its level was reduced significantly.The larger the membrane pore size was,the earlier and the more RNA was detected.ConclusionsThe membrane pore size,the treatment time and the viral level in the internal circuit are contributing factors influencing the transfer of PERV RNA across the membrane in a BAL.

Objective To evaluate the effects of regular follow-up and systematic management in premature infants on early discovering of cerebral palsy and intervention efficacy in early phase. Methods Fifty premature infants with cerebral palsy recruited from early children development outpatient service, were classified as treatment group; 40 premature infants with cerebral palsy recruited from general outpatient service, were classified as control group. The infants of treatment group received systematic follow-up and intervention. while the infants of control group received no systematic management. Age and therapeutic efficacy were compared between the two groups. ResultsThe average age in the treatment group ( [4. 0 ± 1.2] months) was significantly younger than that of the control group ( [7.0 ± 1.4] months) ( P ＜ 0. 05 ). The mental development index ( MDI )and physical development index (PDI) in the treatment group was 91.8 ± 10. 2 and 90. 2 ± 11.2,respectively,which were significantly higher than that of the control group(80. 2 ± 11.7 for MDI and 79. 3 ± 10. 2 for PDI)( P ＜ 0. 05 ) . Conclusion Systematic management could help discover cerebral palsy in premature infants in time and could improve physical and mental development of these infants.

AIM: To study the role of TGF-?/Smad pathway in the development of renal fibrosis in diabetic nephropathy.METHODS: Rats were induced to diabetic nephropathy by using tail intravenous injection of STZ.The expression of TGF-?_1,Smad2/3 protein and mRNA in kidney were examined at 2,4,8 and 16 weeks after STZ induction.CTGF,collagen-Ⅲ,PAI-1 mRNA expression in kidney at 16 weeks of STZ-induced diabetic nephropathy and normal rats were studied by RT-PCR.RESULTS: Weak TGF-?_1,Smad2/3 protein were detected in normal renal tissues while strong TGF-?_1,Smad2/3 staining were observed in renal tissues of diabetic nephropathy(0.057?0.030/0.223?0.040;0.017?0.010/0.153?0.010,respectively,P

The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.

0.05).②There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer,including tumor size and portal vein invasion(P