Objective To analyze the reasons and mechanisms of stereoscopic function abnormality in elder patients after the cataract extraction with intraocular lens implantation. Methods 150 cases with artificial lens implantation were randomly selected.Routine eye examinations were carried out and the associated refractive error and presbyopia were corrected.Bilateral simultaneous visual perception and fusion function were examined.The stereoscopic function was tested using "the stereoscopic function examination diagram"created by Yan Shao-ming.Ninety-seven patients who were found to have unrecover-ed or abnormal stereoscopic function were enrolled for analysis. Results In these 97 cases,53(54.6%)were found to have preexisting eye disorders that could affect visual acuity and binocular single vision before the operation.Macular problem was the most prevalent problem in this group.Twenty-seven(27.8%)patients had complicated with corneal astigmatism,after cataract,paralysing strabismus and diplopia as well as macular edema after the operation.In addition,the contralateral unoperated cataract in 17(17.5%)patients and post-operative anisometropia in 9(9.3%)patients were also the causes of stereoscopic function abnormality.There was no reason could be identified in 8 cases. Conclusions The pre-existing eye disorders before lenses implantation,complications of the operation,contralateral unoperated cataract and anisometropia are all the major factors that affect visual acuity recovery and bilateral stereoscopic function rehabilitation.

Objective To screen a cell line which stably suppresses DAPK expression and to observe the growth characters.Methods Four pairs of shRNA were designed,synthesized and inserted into the pDsRed1-N1-U6 vector.The recombinant plasmids were purified and transfected into PC12 cell.Meanwhile,a pDsRed1-N1-U6 vector was transfected as control.The cell clones were screened by G418,and the stable PC12 cell line was established.DAPK expression was detected by Western blot.MTT method and Flow Cytometry(FCM) assay were used to assess the growth characters of the cell line.Results The shRNAs were transfected into PC12 cell and the cell clones were successfully screened out.Of the four recombinant plasmids,the F2 was the best interfering shRNA.Beyond our expectation,the F1 recombinant plasmid had an enhanced effect on DAPK expression.Conclusion A stable PC12 cell line with stable inhibition of DAPK expression by was established using siRNA expression vectors.

Objective To test serum differentially expressed proteins between early-stage (stage IB-ⅢA) and late-stage (stage Ⅳ) lung cancer patients by proteinchip technology and investigate its clinical value. Methods SELDI-TOF-MS and WCX-2 protein chip were used to detect the serum protein of 30 cases of early stage lung cancer patients and 30 cases of late stage lung cancer patients. The data were analyzed by using Biomarker Wizard software. Results There are ten different proteins in the serum between the two groups of lung cancer patients. Four protein markers 7978, 8139, 15 951 and 16 133 are over expressed and seven protein markers 2867, 6885, 8701, 8840, 13 781 and 13 955 are low expressed in the late group. Conclusion SELDI-TOF-MS proteinchip technology is a convenient, sensitive and high-throughput analysis method which can screen several relatively specific protein markers for late stage lung cancer from the serum samples. This selected protein markers can predict metastasis of lung cancer patients.

Animals ; Basilar Artery ; Brain Ischemia ; pathology ; Brain Stem ; blood supply ; Cats ; Medulla Oblongata ; blood supply

OBJECTIVETo observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant.

METHODSSix mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome.

RESULTSWe found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01).

CONCLUSIONCB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

Animals ; Auditory Cortex ; metabolism ; Auditory Pathways ; metabolism ; Calbindins ; metabolism ; Inferior Colliculi ; metabolism ; Mice ; Mice, Inbred C57BL ; Mutation ; Parvalbumins ; metabolism ; Pons ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics

OBJECTIVETo investigate the effect of extracellular regulating kinase (ERK) signaling pathway on the secretion of gamma-aminobutyric acid (GABA) in cultured rat hippocampal neurons induced by stromal cell derived factor-1 (SDF-1).

METHODSThe hippocampal neurons of newborn SD rats were cultured and identified in vitro; the phosphorylation level of ERK1/2 was examined by Western blot; ELISA was used to detect the effect of PD98059, a ERK1/2 specific blocker on GABA secretion of cultured hippocampal neurons and Western blot were adopted to measure the protein expression levels of glutamate decarboxylase (GAD65/67) and gamma aminobutyric acid transporter (GAT); after blocking ERK1/2 signaling pathway with PD98059; RT-PCR was used to detect the mRNA expression levels of GAT-1 and GAD65 after treated with PD98059.

RESULTSThe levels of ERKl/2 phosphorylation were increased significantly by SDF1 acting on hippocampal neurons, and CX-CR4 receptor blocker AMD3100, could inhibit SDF-1 induced ERK1/2 activation; SDF-1 could inhibit the secretion of GABA in cultured hippocampal neurons, and ERK1/2 specific inhibitor PD98059, could partly reverse the inhibition of GABA secretion by SDF-1. The effects of SDF-1 on cultured hippocampal neurons was to decrease the mRNA genesis of glutamic acid decarboxylase GAD65 and GABA transporter GAT-1, besides, ERK inhibitor PD98059 could effectively flip the effect of SDF-1. The results of Western blot showed that SDF-1 could inhibit the protein expression of GAT-1 and GAD65/67 in hippocampal neurons and the inhibition of GAT-1 and GAD65/67 protein expression could be partially restored by ERK1/2 blocker.

CONCLUSIONSDF-1 acts on the CXCR4 of hippocampal neurons in vitro, and inhibits the expression of GAD by activating the ERK1/2 signaling pathway, and this may represent one possible pathway of GABA secretion inhibition.

Animals ; Blotting, Western ; Cells, Cultured ; Chemokine CXCL12 ; pharmacology ; Flavonoids ; pharmacology ; Glutamate Decarboxylase ; metabolism ; Hippocampus ; cytology ; MAP Kinase Signaling System ; Neurons ; metabolism ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptors, CXCR4 ; metabolism ; gamma-Aminobutyric Acid ; secretion

Objective To estimate reliability of magnetic resonance elastography (MRE) in measuring liver stiffness of volunteers and patients with chronic liver disease and to assess inter-rater consistency.Methods MRE was performed on a 3.0 T scanner in all subjects,including 24 volunteers (control group) and 64 patients with liver disease (chronic liver disease group).Liver stiffness was measured blindly by two raters.The pathological fibrosis score was applied as a standard reference for liver fibrosis in 22 patients.The intraclass correlation coefficient (ICC) was used to evaluate inter-rater reliability.The differences of liver stiffness between two groups were evaluated using non-parametric MannWhitney U test.Spearman analysis was used to analyze the correlation between fibrosis stages and liver stiffness.Results The intraclass correlation coefficient of liver stiffness was perfect (ICC =0.99,P ＜ 0.01)between two raters.There was significant difference of mean stiffness between control group and patient group (U =90.5,P ＜0.01) with(2.35 ±0.34) kPa and(4.17 ± 0.47) kPa,respectively.The correlation between fibrosis stage (3,3,5,5 and 6 patients in fibrosis stage S0,S1,S2,S3 and S4) and stiffness (2.13,3.25,3.82,5.45 and 7.35 kPa) was very strong (r =0.96,P ＜0.01).Conclusion MRE is a reliable and promising tool to measure liver stiffness and to assess liver fibrosis.

BACKGROUND:The mycobacterium tuberculosis heat shock protein 10 exerts effects on the osteoclasts by in vitro mouse cranium experiment,
OBJECTIVE:To investigate the effect and mechanism of recombinant mycobacterium tuberculosis heat shock protein 10 (CPN10) on the differentiation of osteoclasts in the in vitro culture system that induces osteoclast differentiation.
METHODHuman macrophage colony-stimulating factor-dependent adhesive blood mononuclear cells were divided into four groupreceptor activator for nuclear factor-κB ligand (RANKL)+CPN10 (1 mg/L), RANKL, CPN10 (1 mg/L), and negative control (complete culture medium). Monocytes were resuspended in a-MEM medium containing macrophage colony-stimulating factor, and were cultured in each group for 7, 14, 21 days. The morphology, quantity and bone resorption area of osteoclasts were examined by tartrate-resistant acid phosphatase (TRAP) staining. The expressions of NFATc1 and c-Fos gene and protein were also detected.
RESULTS AND CONCLUSION:In negative control group, no TRAP-positive multinucleated osteoclasts generated, while in the other groups, TRAP-positive multinucleated osteoclasts differentiated and formed the lacunae in the smal bone grinding. The number of osteoclasts formation and resorption in CPN10 group were significantly lower than that in RANKL+CPN10 group. The expression of NFATc1 and c-Fos in the negative control group C was significantly lower than that of RANKL+CPN10 group and CPN10 group. However, CPN10 expressed NFATc1 and c-Fos protein, which was significantly lower than RANKL+CPN10 group. CPN10 is involved in the formation of osteoclasts, and the mechanism is related with the upregulation of NFATc1, c-Fos expression.

BACKGROUND:Mycobacterium tuberculosis heat shock protein 10 (r-Mt cpn10) is one of the main factors that cause bone tuberculosis dissolution and absorption as wel as inhibits the proliferation of osteoblasts. Receptor activator of nuclear factor kappa B ligand and osteoprotegerin are the important factors influencing bone metabolism.
OBJECTIVE:To observe the effect of r-Mt cpn10 on human osteoblast proliferation, alkaline phosphatase secretion, expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA. METHODS:Human bone marrow stromal cel s were induced to differentiate into osteoblasts, and osteoblasts at passage 3 were cultured with various concentrations of r-Mt cpn10 (0.1, 1, 10 mg/L). Osteoblasts cultured without r-Mt CPN10 were assigned as controls.
RESULTS AND CONCLUSION:MTT assay results showed that, compared with control group, r-Mt cpn10 at different concentrations inhibited osteoblast proliferation and alkaline phosphatase secretion (P<0.05). RT-PCR analysis showed that, r-Mt cpn10 at different concentrations increased receptor activator of nuclear factor-kappa B ligand mRNA expression (P<0.01), and inhibited osteoprotegerin mRNA expression in a concentration-dependent manner (P<0.01). 10 mg/L r-Mt cpn10 exhibited the strongest effect (P<0.01). The r-Mt cpn10 can inhibit osteoblast proliferation and alkaline phosphatase activity, and it may influence bone metabolism by regulating the expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA.

Objective To investigate the clinical and imaging features in patients with vertebrobasilar dolichoectasia (VBD) through a comparative study in patients with ischemic stroke with or without VBD.Methods The patients with acute cerebral infarction were divided into either a VBD group or a non-VBD group according to magnetic resonance angiography.The VBD group was further divided into an anterior circulation infarction subgroup and a posterior circulation infarction subgroup.The cardiovascular risk factors,the diameter of basiar artery (BA),bifurcation height,and horizontal displacement were compared in all groups.Results A total of 269 patients with acute cerebral infarction were included,28 had VBD,accounting for 10.41％ of the patients with acute cerebral infarction during the same period.The proportion of male patients (78.6％ vs.66.8％ ;x2 =4.392,P =0.036),age (70.38 ± 10.58 years vs.62.86 ± 12.20 years; t =2.870,P =0.009),and the proportion of hypertension (89.3％ vs.47.7％ ; x2 =17.367,P =0.000) in the VBD group were significantly higher than those in the non-VBD group.The multivariate logistic regression analysis showed that the advanced age (odds ratio [OR] 1.248,95％ confidence interval [CI] 1.137-1.371; P=0.000),hyperglycemia (OR 1.599,95％ CI 1.181-2.164; P =0.002),hypertension (OR 1.251,95％ CI 1.020-1.534; P =0.032) and increased triglyceride level (OR 1.876,95％ CT 1.021-3.445; P =0.043) were the independent risk factors for VBD,while female gender (OR 0.133,95％ CI 0.024-0.735; P =0.021) was the independent protective factor for VBD.Of the 28 cerebral infarction patients with VBD,9 had anterior circulation infarction and 19 had posterior circulation infarction.There were significant differences in BA diameter ([5.40 ± 0.49] cm vs.[6.00 ± 0.77] cm; t =2.046,P =0.041),and the proportions of high score in bifurcation height (x2 =6.768,P =0.037) and horizontal displacement (x2 =5.241,P =0.042) between the 2 groups (all P ＜0.05).The multivafiate logistic regression analysis showed that the BA bifurcation height was an independent risk factor for posterior circulation infarction (OR 1.347,95％ CI 1.069-2.457; P =0.038) in patients with VBD.Conclusions VBD accounted for 10.41％ of the patients with acute cerebral infarction during the same period.Advanced age,hyperglycemia,hypertension and increased triglyceride level were the independent risk factors for VBD.Female gender was the independent protective factors for VBD,and the BA bifurcation height was an independent risk factor for VBD occurring posterior circulation infarction.