OBJECTIVE Neuroinflammation plays a critical role in neurodegenerative disorders, although the inflammation may not the initiating factor. Parkinson disease (PD) is characterized patho?logically by the accumulation of alpha synuclein (α-syn) and the loss of the dopamine (DA) neurons in the substantia nigra (SN), which has been reported to be induced by the stereotaxic injection of lipopolysaccharide (LPS) to the SN region in rodents. This study is to investigate the therapeutic benefit of the inhibition of miR-873 in PD. METHODS Rats received the right-unilaterally injection with concentrated LV-sponge or LV-EGFP 3 d before LPS treatment, 7 or 14 d after LPS treatment. The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 21 d after LPS injection. The regulation of miR-873 on the genes related with cholesterol transport and inflammation was assayed in SH-SY5Y cells and U251 cells. RESULTS TLR4-MyD88 signaling pathway was involved the regulation of miR-873 by LPS. The luciferase assay showed that HMGCR, ABCA1 and A20 were down- stream genes of miR- 873. The transfection of miR- 873 decreased the cholesterol levels in cell membrane, but increased in lysosome in SH-SY5Y cells. Compared with the control SH-SY5Y cells, cholesterol levels were higher in lysosome with α-synuclein overexpression or LPS treatment. The transfection of miR-873 increased the α-syn levels in lysosome in cells with α-synuclein overexpression. The loss of dopaminergic neuorns induced by LPS was significantly respectively decreased by 22.8%, 35.6% and 57% after the inhibition of miR-873 at 3 d before LPS treatment, 7 or 14 d after LPS treatment. Compared with LPS-treated group, the number of the rotation of rats was decreased by 60.4%, 33.5% and 13.2% after the inhibition of miR-873 at 3 d before LPS treatment, 7 or 14 d after LPS treatment. The inhibition of miR-873 significantly decreased accumulation of α-syn. The mRNA levels of HMGCR, ABCA1 and A20 in SN were decreased by LPS treatment, which was attenuated by the injection of LV- sponge. CONCLUSION The selective regulation of miR- 873 can protect the dopaminergic neurons from the LPS-induced damage. The inhibition of miR-873 can attenuate the relocation of cholesterol in lysosome and the accumulation of α-syn in neurons induced by LPS via the regulation of HMGCR, ABCA1 and A20.

OBJECTIVE CYP2D is one of the most abundant subfamily of CYPs in the brain, especially in the cerebellum. Brain CYP2D is responsible for the metabolism of endogenous neurotransmitters such as tyramine and serotonin. Our previous studies have shown brain CYP2D can be regulated by exogenous and endogenous substances with tissue- specificity. The purpose of this study is to examine the effects of cerebral CYP2D on the mice behavior and the regulatory mechanism of brain CYP2D by growth hormone. METHODS Mice received the stereotaxic injection with CYP2D inhibitor quinine in deep cerebellar nuclei of cerebellum. The animals were tested with rotarod apparatus, balance beam, water maze, elevated plus maze and open field. The changes in CYP2D22, PPARαand PPARγ in brain regions and liver were assayed in male growth hormone receptor knockout mice, SH-SY5Y cells and HepG2 cells. RESULTS The inhibition of cerebellum CYP2D significantly affected the spatial learning and exploring ability of mice. Compared with WT mice, CYP2D expression was lower in brain regions from GHR(-/- ) male mice; however, hepatic CYP2D level was similar. Pulsatile GH decreased PPARα mRNA level, and increased mRNA levels of CYP2D6 and PPARα in SH- SY5Y cells. In HepG2 cells, pulsatile GH resulted in decreases in PPARα and PPARγ mRNA levels, but not CYP2D6. PPARα inhibitor induced CYP2D6 mRNA and protein by 1.32-fold and 1.43-fold in SH-SY5Y cells. PPARγ inhibitor decreased CYP2D6 mRNA and protein by 74.76% and 40.93%. PPARα agonist decreased the level of CYP2D22 mRNA in liver and cerebellum, while PPARγ agonist rosiglitazone resulted in diametrically increases. The luciferase assay showed that PPARγ actived the CYP2D6 gene promoter while PPARα inhibited its function. Pulsatile GH declined the binding of PPARα with CYP2D6 promoter by 40%, promoted the binding of PPARγ with CYP2D6 promoter by approximate 60%. The levels of brain and liver PPARα expression in male GHR(-/- ) mice is obviously higher than those in WT mice. The level of PPARγ in male GHR(-/- ) mice was decreased in the frontal cortex and hippocampus, while remained stable in the cerebellum and striatum; meanwhile, PPARγ was increased in the liver. CONCLUSION Brain CYP2D may be involved in learning and memory functions of central system. Masculine GH secretion altered the PPARs expression and the binding of PPARs to CYP2D promoter, leading to the elevated brain CYP2D in a tissue- specific manner. Growth hormone may specifically alter the metabolic and synthetic of important endogenous substances in the central nervous system (such as serotonin) through the specific regulation of brain CYP2D expression.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; DNA Fragmentation ; drug effects ; Drug Resistance, Neoplasm ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Inhibitor of Apoptosis Proteins ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; pharmacology ; Ovarian Neoplasms ; genetics ; pathology ; ultrastructure ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effect of PD98059 on the proliferation and cell cycle of rat hepatic stellate cells (HSCs) stimulated by acetaldehyde and explore its mechanism.

METHODSRat HSCs stimulated by acetaldehyde were incubated with different concentrations of PD98059. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle was analysed by flow cytometry. The mRNA of cyclin D1 and CDK4 were examined by RT-PCR.

RESULTS20, 50, 100 micromol/L PD98059 could significantly inhibit the proliferation of HSCs stimulated by acetaldehyde in a does-dependent manner (0.109+/-0.020, 0.081+/-0.010 and 0.056+/-0.020 vs 0.146+/-0.030, F=31.385, P<0.05) and provoke G0/G1 phase arrest of HSCs stimulated by acetaldehyde in a does-dependent manner (61.9%+/-6.3%, 64.1%+/-3.3% and 70.9%+/-4.8% vs 55.2%+/-4.4%, F=16.402, P<0.05). 50, 100 micromol/L PD98059 could markedly inhibit cyclin D1 mRNA expression of HSC stimulated by acetaldehyde (0.56+/-0.04 and 0.46+/-0.03 vs 0.65+/-0.07, F=68.758, P<0.05) and CDK4 mRNA expression (0.39+/-0.07 and 0.33+/-0.05 vs 0.50+/-0.06, F=29.406, P<0.05).

CONCLUSIONThe Erk signal transduction pathway plays an important role in regulating the proliferation and cell cycle of rat hepatic stellate cells stimulated by acetaldehyde, which may be partly related to its regulative effect on the expression of cyclin D1 gene and CDK4 gene

Acetaldehyde ; pharmacology ; Animals ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinases ; metabolism ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Hepatocytes ; drug effects ; Proto-Oncogene Proteins ; Rats

Objective To evaluate the effect of PTH gene polymorphisms on bone mineral density (BMD) at multiple skeletal sites in normal females.Methods PTH gene phenotype was determined by PCR-RFLP of restriction enzyme Bst BⅠin 596 females aged (46.3?13.7) years (20-80 years),and PCR products with or without enzymolytic site were considered as genotype B or genotype b respectively.BMDs of the anteropesterior spine (AP) and supine lateral spine (Lat) of lumbar vertebrae (L_1-L_4),femoral neck (FN),total hip (T-hip), Ward's triangle (Ward),Trochanter (Troch),forearm [radius+ulna ultradistal (RUUD) and total area of radius + ulna (RUT) ] were measured by DEXA (QDR4500A).Results (1) Hardy-Weinberg equilibrium was evident for PTH polymorphisms.The frequencies of genotype were BB 0.784,Bb 0.208,bb 0.008 and frequencies of alleles B,b were 0.888 and 0.112 respectively in 596 normal females.Frequencies of BB,Bb,bb genotypes were 0.781,0.210,and 0.009 respectively in 347 postmenopausal women and their frequencies of alleles B,b were 0.886,0.114.No significant difference was found between post- and premenopausal women in genotype frequen- cy.(2) Comparing their BMDs of AP,Lat,FN,T-hip,Ward,Troch,RUUD and RUT,there was no significant difference between BB and Bb genotypes of pre- and postmenopansal women groups.(3) Logistic regression analysis failed to show any statistical difference between normal and osteoporosis women with regard to PTH phenotype.Conclusion PTH gene polymorphism has little effect on BMD in normal females.

OBJECTIVE: To investigate the relationship between estrogen receptor (ER) gene polymorphisms and HBV-induced cirrhosis. METHODS: Xba I and Pvu II polymorphisms of ER gene were analyzed in 98 patients with HBV-induced cirrhosis, 72 patients with chronic hepatitis B, and 84 healthy controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. RESULTS: The frequencies of Pp genotype and P allele of ER gene in patients with HBV-induced cirrhosis were higher than those in patients with chronic hepatitis B and controls, while the frequencies of pp genotype and p allele of ER gene in patients with HBV-induced cirrhosis were lower than those in patients with chronic hepatitis B and controls (P < 0.05). The risk of HBV-induced cirrhosis occurrence in carriers with Pp and pp genotype was 2.23 folds than that in carriers with pp genotype. There was no significant difference in frequencies of allele and genotype in Xba I polymorphism among the HBV-induced cirrhosis group, the chronic hepatitis B group, and the control group (P > 0.05). CONCLUSION Pp genotype and P allele might be the susceptibility gene for HBV-induced cirrhosis while pp genotype and p allele might be the protective gene.
Adolescent ; Adult ; Aged ; Alleles ; Female ; Genotype ; Hepatitis B, Chronic ; complications ; genetics ; Humans ; Liver Cirrhosis ; etiology ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Receptors, Estrogen ; genetics

Objective To investigate the correlation between the changes of serum S100B protein level in acute phase and the scores of mini-mental state examination (MMSE) in patients with brain concussion, and evaluate the role of serum S100B protein level in the prognosis of cognition disorders after brain concussion. Methods The serum S100B protein level was determined by an enzyme linked immunosorbent assay (ELISA) in 126 cases of brain concussion 6 and 12 h, and 3 d after admission, and these data were compared with those in 96 cases of moderate head trauma without transitory loss of consciousness (admitted to our hospital at the same period, control group). MMSE was performed 1 and 14 d and 3 months after injury, and the correlation between the changes of serum S100B level in acute phase and MMSE scores was observed. Results As compared with that in control group,the serum S100B protein level in patients with brain concussion was significant higher at 6 and 12 h after admission(P＜0.05). The serum S100B protein level at 6 h, but not at 12 h and 3 d after admission, was closely associated with the MMSE scores 1 and 14 d and 3 months after injury. Conclusion Early elevation of S100B within 6 h of admission in patients with brain concussion, obviously correlating with cognitive impairment, may serve as an important prognostic marker in predicting clinical outcome of cognition disorders after brain injury.

Objective To investigate the effective treatments of combined injury with craniocerebral firearm wound in dogs immersed by seawater under maritime environment. Methods Models of combined injury with craniocerebral firearm wound, including craniocerebral gunshot wound,open chest injury, open abdominal injury, open trauma of extremities and burn injury, were established in 60 healthy adult mongrel dogs. Animal models after being wounded were immersed by the seawater for 30 min, and then, they were equally randomized into conventional treatment group and comprehensive treatment group; 30 dogs in the conventional treatment group were given routine treatment and the other 30 dogs in the comprehensive treatment group were given lukewarm glucose liquid, β-aescin, naloxone hydrochloride, levofloxacin and re-warming treatments besides the conventional treatment. Transcranial Doppler ultrasound, blood gas analysis, measurement of plasma osmotic pressure and intracranial pressure (ICP) monitoring were performed on the dogs of the 2 groups; and the treatment efficacy of the 2groups were compared. Results Low incidence rate of brain vasospasm was noted and TCD indicated that blood flow speed approached normal in the comprehensive treatment group 3 h after the treatment.The plasma osmotic pressure and the indicators of metabolic acidosis reached normal levels in the comprehensive treatment group 12 h after the treatment. The ICP significantly decreased in the comprehensive treatment group 24 h after the treatment. Survival rate in the comprehensive treatment group (70%) was significantly higher as compared with that in the conventional treatment group (53%)7 d after the treatment (P＜0.05). All the indexes in the comprehensive treatment group were better than those in the conventional treatment group (P＜0.05) Conclusion Early infusion of lukewarm hypotonic solution can significantly reduce the osmotic pressure, correct the electrolyte balance, help the re-warming and prolong the survival rate. Naloxone possesses protective effect on brain. The β-aescine sodium can diminish viscosity, slow down brain edema progress, obviously reduce ICP and improve brain tissue oxygen metabolism. In a word, comprehensive treatment in effective in treating combined injury with craniocerebral firearm wound.

Objective To demonstrate the effect of Pioglitazone hydrochloride on spatial memory ability in diabetes rats via improving the level of hippocampal cholinergic neurostimulating peptide (HCNP) in the hippocampus tissues. Methods Twenty-four 12-week old male SD rats were equally randomized into control group (Con),diabetes group (DM),DM+Pioglitazone hydrochloride treatment group (DP).Rats of the DM and DP groups were fed with high fat diet,and rats of the DP group were also performed intragastric administration of 10 mg/(kg· d) Pioglitazone hydrochloride daily.Morris water maze test was performed on the rats enjoyed successful model making to detect their spatial memory ability; the level of HCNP precursor protein (HCNP-pp) was measured by Western blotting so as to measure the level of HCNP; and the relative mRNA contents of HCNP-pp, choline acetyltransferase (ChAT),muscarinic receptor 1 (M1R) and α7 type nicotinic receptor (α7NR) genes were measured by reverse transcription PCR. Results The escape latency in rats of the con and DP groups was significantly shortened as compared with that in the DM group (P＜0.05).As compared with that in the Con and DP groups, the relative amount of HCNP-pp in DM group was higher in protein and mRNA levels (P＜0.05); however,the relative amount of HCNP in the DM group was obviously lower than that in the Con and DP groups (P＜0.05).And the mRNA ChAT,M1R and α7NR levels in the Con and DP groups were significantly higher than those in the DM group (P＜0.05). Conclusion Pioglitazone hydrochloride can accelerate the lysis of HCNP-pp in the hippocampus to induce the increased level of HCNP,further making the cholinergic nerve receptors activation,which maybe one of the mechanisms to improve the ability of spatial memory in eldly rats with type 2 diabetes.