[Changes in Plasma Amyloid-ß Level and Their Relationship With White Matter Microstructure in Patients With Mild Cognitive Impairment].

Objective To investigate the changes in plasma amyloid-ß (Aß) level and their relationship with white matter microstructure in the patients with amnesic mild cognitive impairment(aMCI) and vascular mild cognitive impairment (vMCI).Methods A total of 36 aMCI patients,20 vMCI patients,and 34 sex and age matched healthy controls (HC) in the outpatient and inpatient departments of the First Affiliated Hospital of Anhui Medical University were enrolled in this study.Neuropsychological scales,including the Mini-Mental State Examination,the Montreal Cognitive Assessment,and the Activity of Daily Living Scale,were employed to assess the participants.Plasma samples of all the participants were collected for the measurement of Aß42 and Aß40 levels.All the participants underwent magnetic resonance scanning to obtain diffusion tensor imaging (DTI) data.The DTI indexes of 48 white matter regions of each individual were measured (based on the ICBM-DTI-81 white-matter labels atlas developed by Johns Hopkins University),including fractional anisotropy (FA) and mean diffusivity (MD).The cognitive function,plasma Aß42,Aß40,and Aß42/40 levels,and DTI index were compared among the three groups.The correlations between the plasma Aß42/40 levels and DTI index of aMCI and vMCI patients were analyzed.Results The Mini-Mental State Examination and the Montreal Cognitive Assessment scores of aMCI and vMCI groups were lower than those of the HC group (all P<0.001).There was no significant difference in the Activity of Daily Living Scale score among the three groups (P=0.654).The plasma Aß42 level showed no significant difference among the three groups (P=0.227).The plasma Aß40 level in the vMCI group was higher than that in the HC group (P=0.014),while it showed no significant difference between aMCI and HC groups (P=1.000).The plasma Aß42/40 levels in aMCI and vMCI groups showed no significant differences from that in the HC group (P=1.000,P=0.105),while the plasma Aß42/40 level was lower in the vMCI group than in the aMCI group (P=0.016).The FA value of the left anterior limb of internal capsule in the vMCI group was lower than those in HC and aMCI groups (all P=0.001).The MD values of the left superior corona radiata,left external capsule,left cingulum (cingulate gyrus),and left superior fronto-occipital fasciculus in the vMCI group were higher than those in HC (P=0.024,P=0.001,P=0.003,P<0.001) and aMCI (P=0.015,P=0.004,P=0.019,P=0.001) groups,while the MD values of the right posterior limb of internal capsule (P=0.005,P=0.001) and left cingulum (hippocampus) (P=0.017,P=0.031) in the aMCI and vMCI groups were higher than those in the HC group.In the aMCI group,plasma Aß42/40 level was positively correlated with FA of left posterior limb of internal capsule (r=0.403,P=0.015) and negatively correlated with MD of the right fonix (r=-0.395,P=0.017).In the vMCI group,plasma Aß42/40 level was positively correlated with FA of the right superior cerebellar peduncle and the right anterior limb of internal capsule (r=0.575,P=0.008;r=0.639,P=0.002),while it was negatively correlated with MD of the right superior cerebellar peduncle and the right anterior limb of internal capsule (r=-0.558,P=0.011;r=-0.626,P=0.003).Conclusions Plasma Aß levels vary differently in the patients with aMCI and vMCI.The white matter regions of impaired microstructural integrity differ in the patients with different dementia types in the early stage.The plasma Aß levels in the patients with aMCI and vMCI are associated with the structural integrity of white matter,and there is regional specificity between them.

Clinical Utility of the Serum Level of Lipoprotein-Related Phospholipase A2 in Acute Ischemic Stroke With Cerebral Artery Stenosis.

We aimed to study the clinical utility of serum lipoprotein-associated phospholipase A2 (Lp-PLA2) in acute ischemic stroke (AIS) with cerebral artery stenosis (CAS). We included 200 AIS patients and 90 healthy controls in this study. AIS patients were classified into three subgroups depending on the severity of CAS. They were also classified based on the stability of the carotid plaques. Spearman correlation analysis was performed to determine the correlation relationship between the level of Lp-PLA2 and neurologic injury. Binary logistic regression analysis was performed to determine the independent risk factors for AIS. Receiver operating characteristic (ROC) analysis was performed to assess the diagnostic value of Lp-PLA2 for AIS and for the degree of CAS. We found that the serum level of Lp-PLA2 in AIS patients was significantly higher than that in the control group. Lp-PLA2 was further identified as an independent risk factor for AIS (p = 0.001, OR = 1.057). In addition, serum Lp-PLA2 level was the highest in AIS patients with severe CAS or occlusion. Lp-PLA2 level was higher in AIS patients with unstable plaques and in AIS patients with moderate to severe neurological injury. Lp-PLA2 level was positively correlated with National Institutes of Health Stroke Scale (NIHSS) score (r = 0.335, p = 0.001). We found that the optimal cut-off value for Lp-PLA2 level was 123.365 ng/ml, at which the sensitivity and specificity for the diagnosis of ACI were 74.5 and 86.7%, respectively, and the area under ROC curve (AUC) was 0.892. Similarly, the optimal value for Lp-PLA2 level was 136.46 ng/ml, at which the sensitivity and specificity for the diagnosis of the presence of moderate to severe artery stenosis or occlusion were 79.6 and 95.2%, respectively, and the AUC was 0.938. The ROC curve indicated that serum Lp-PLA2 level has an excellent diagnostic value for AIS and severe stenosis. Based on these results we conclude that Lp-PLA2 could be a potential biomarker to complement the current imaging methods in the prediction and diagnosis of AIS. An elevated Lp-PLA2 level is also correlated with carotid plaque instability, severe neurological injury and cerebrovascular stenosis. Future longitudinal studies are needed to determine whether there is a causative relationship between Lp-PLA2 and AIS.

Contribution of Inflammation and Hypoperfusion to White Matter Hyperintensities-Related Cognitive Impairment.

White matter hyperintensities (WMHs) of presumed vascular origin are one of the most important neuroimaging markers of cerebral small vessel disease (CSVD), which are closely associated with cognitive impairment. The aim of this study was to elucidate the pathogenesis of WMHs from the perspective of inflammation and hypoperfusion mechanisms. A total of 65 patients with WMHs and 65 healthy controls were enrolled in this study. Inflammatory markers measurements [hypersensitive C-reactive protein (hsCRP) and lipoprotein-associated phospholipase A2 (Lp-PLA2)], cognitive evaluation, and pseudocontinuous arterial spin labeling (PCASL) MRI scanning were performed in all the subjects. The multivariate logistic regression analysis showed that Lp-PLA2 was an independent risk factor for WMHs. Cerebral blood flow (CBF) in the whole brain, gray matter (GM), white matter (WM), left orbital medial frontal gyrus [MFG.L (orbital part)], left middle temporal gyrus (MTG.L), and right thalamus (Tha.R) in the patients was lower than those in the controls and CBF in the left triangular inferior frontal gyrus [IFG.L (triangular part)] was higher in the patients than in the controls. There was a significant correlation between Lp-PLA2 levels and CBF in the whole brain (R = -0.417, p < 0.001) and GM (R = -0.278, p = 0.025), but not in the WM in the patients. Moreover, CBF in the MFG.L (orbital part) and the Tha.R was, respectively, negatively associated with the trail making test (TMT) and the Stroop color word test (SCWT), suggesting the higher CBF, the better executive function. The CBF in the IFG.L (triangular part) was negatively correlated with attention scores in the Cambridge Cognitive Examination-Chinese Version (CAMCOG-C) subitems (R = -0.288, p = 0.020). Our results revealed the vascular inflammation roles in WMHs, which may through the regulation of CBF in the whole brain and GM. Additionally, CBF changes in different brain regions may imply a potential role in the modulation of cognitive function in different domains.

Combined dimercaptosuccinic acid and zinc treatment in neurological Wilson's disease patients with penicillamine-induced allergy or early neurological deterioration.

The clinical data of safety and efficacy of a combined treatment with dimercaptosuccinic acid (DMSA) and Zinc with 2 years' follow-up in 60 neurological Wilson's disease (WD) patients was retrospectively analyzed. All the patients included in the present study were newly diagnosed and initialized with D-penicillamine (DPA) treatment but were found to have either neurological deterioration or allergy, and their treatment was switched to a combined treatment of DMSA and Zinc. Fifty-one patients (85%) had the neurological symptoms improved 1 and 2 years after treatment, 7 (11.67%) experienced a stable neurological condition, and 2 (3.33%) suffered deterioration of neurological symptoms. No early neurological deterioration was observed in all patients. Twenty-five percent patients experienced mild adverse reactions which did not require a discontinuation of the DMSA and Zinc treatment. Our study confirmed the safety and efficacy of the combined DMSA and Zinc therapy as an initial and probably long-term treatment in neurological WD patients.

Acute viral encephalitis associated with human parvovirus B19 infection: unexpectedly diagnosed by metagenomic next-generation sequencing.

We report here a case of a 17-year-old boy with viral encephalitis associated with human parvovirus B19 who presented consciousness disturbance, left hemiparesis, and focal neurologic signs. The diagnosis was based on the specific sequence reads corresponding to human parvovirus B19 (PVB19) in a CSF sample as analyzed by metagenomic next-generation sequencing (mNGS). Thus, PVB19 should be considered in the differential diagnosis of encephalitis and encephalopathy of unknown etiology. The introduction of mNGS into the diagnostic protocol of neuropathies, especially for those undiagnosed, could interrogate all genetic information in a biologic sample and facilitate the identification of the etiological agent.

Acute onset neurological symptoms in Wilson disease after traumatic, surgical or emotional events: A cross-sectional study.

Acute onset neurological symptoms evoked by traumatic, surgical, or emotional events in Wilson disease (WD) have never been reported and its clinical characteristics are unclear.We aimed to summarize the clinical characteristics of a special WD whose neurological symptoms acutely developed after traumatic, surgical, or emotional events.Retrospective pilot study.Thirty-one patients who had acute onset neurological symptom as an initial presentation of WD or a new presentation of hepatic WD after mild trauma, surgery, or emotional events were retrospectively studied. All patients were followed for half to 1 year after regular anti-copper treatment.The averaged latency for neurological symptom presentation was 2.79 ±â€Š1.21 hours. The most frequent neurological symptoms were tremor (74%) and basal ganglia (BG) lesions were detected on magnetic resonance imaging in all patients. Lesions in other regions were much less frequently detected. Neurological symptom score and its recovery after treatment were correlated with lesion location: BG area and BG plus other brain areas. Neurological symptoms improved in 21 patients who received timely anti-copper treatment but continued to deteriorate in 6 patients who did not accept regular anti-copper treatment for delayed diagnosis.A diagnosis of WD should be considered when adolescents or adults experience acute presentation of extrapyramidal systems after traumatic, surgical, or emotional stimulation. Timely anti-copper therapy usually gives rise to an excellent prognosis.

Synthesis of high performance Ni3C-Ni decorated thermally expanded reduced graphene oxide (TErGO/Ni3C-Ni) nanocomposite: A stable catalyst for reduction of Cr(VI) and organic dyes.

A high performance thermally expanded reduced graphene oxide (TErGO) nanocomposite decorated with Ni3C-Ni nanoparticles (TErGO/Ni3C-Ni) has been successfully synthesized by using a facile and eco-friendly approach. The morphology, textural features, surface composition and stability of TErGO/Ni3C-Ni nanocomposite are investigated by various physicochemical characterizations which revealed the uniform dispersion of crystalline metal nanoparticles inside TErGO matrix. The composite has been exhibited a large surface area and pore volume of 121 m2 g-1 and 0.791 cm3 g-1, respectively. The TErGO/Ni3C-Ni exhibited remarkable catalytic performance surpassing most metal-based catalysts with various kind of support matrices reported in recent literature. The reduction of Cr(Ⅵ) to Cr(Ⅲ) was achieved within 1 min with an excellent rate constant of 2.74 min-1 and phenomenally higher specific removal rate (SRR) of 0.29 mg Cr(VI) min-1. mg-1 of TErGO/Ni3C-Ni. While it also proved an excellent reducing catalyst for organic dyes via NaBH4 with full reduction achieved within 30 s. Moreover, as prepared nanocatalyst possesses excellent stability and recyclability with easy magnetic separation.

Spectrum of SLC20A2, PDGFRB, PDGFB, and XPR1 mutations in a large cohort of patients with primary familial brain calcification.

Primary familial brain calcification (PFBC) is a rare neurodegenerative disorder with four causative genes (SLC20A2, PDGFRB, PDGFB, and XPR1) that have been identified. Here, we aim to describe the mutational spectrum of four causative genes in a series of 226 unrelated Chinese PFBC patients. Mutations in four causative genes were detected in 16.8% (38/226) of PFBC patients. SLC20A2 mutations accounted for 14.2% (32/226) of all patients. Mutations in the other three genes were relatively rare, accounting for 0.9% (2/226) of all patients, respectively. Clinically, 44.8% of genetically confirmed patients (probands and relatives) were considered symptomatic. The most frequent symptoms were chronic headache, followed by movement disorders and vertigo. Moreover, the total calcification score was significantly higher in the symptomatic group compared to the asymptomatic group. Functionally, we observed impaired phosphate transport induced by seven novel missense mutations in SLC20A2 and two novel mutations in XPR1. The mutation p.D164Y in XPR1 might result in low protein expression through an enhanced proteasome pathway. In conclusion, our study further confirms that mutations in SLC20A2 are the major cause of PFBC and provides additional evidence for the crucial roles of phosphate transport impairment in the pathogenies of PFBC.

Cerebral Venous Sinus Thrombosis following Diagnostic Curettage in a Patient with Uterine Fibroid.

Cerebral venous sinus thrombosis (CVST) is a relatively rare cerebrovascular disease, of which the risk has been documented in patients with numerous conditions. However, CVST has never been previously described in association with the use of a diagnostic curettage in patient with uterine fibroid. Herein, we described a 43-year-old woman who presented with recurrent convulsive seizures and severe and progressive headache 1 day after a diagnostic curettage of the uterus, which was confirmed to be uterine fibroid pathologically later, and her condition subsequently progressed to confusion. Brain magnetic resonance imaging (MRI) revealed an acute extensive thrombosis of the left transverse and sigmoid sinus and the ipsilateral cerebellum infarction. Evaluation for primary thrombophilia revealed that an iron deficiency anemia (IDA) due to the fibroid bleeding induced menorrhagia together with a diagnostic curettage might be the sole hypercoagulable risk factor identified. Treatment with anticoagulation led to full recovery of her symptoms and recanalization of the thrombosis was proven on magnetic resonance venography (MRV) 2 months later. We suggest that CVST should be recognized as a potential complication related to this diagnostic technique, especially in patient with IDA. The early diagnosis and timely treatment would be of significance in improving the prognosis of this potentially lethal condition.

Linear headache: a recurrent unilateral head pain circumscribed in a line-shaped area.

BACKGROUND: A headache circumscribed in a line-shaped area but not confined to the territory of one particular nerve had ever been described in Epicrania Fugax (EF) of which the head pain is moving and ultrashort. In a 25-month period from Feb 2012 to Mar 2014, we encountered 12 patients with a paroxysmal motionless head pain restricted in a linear trajectory. The head pain trajectory was similar to that of EF, but its all other features obviously different from those of EF. We named this distinctive but undescribed type of headache linear headache (LH). METHODS: A detailed clinical feature of the headache was obtained in all cases to differentiate with EF, trigeminal autonomic cephalalgias (TACs) and cranial neuralgia. Similarities and differences in clinical features were compared between LH and migraine. RESULTS: The twelve LH patients (mean age 43.9 ± 12.2) complained of a recurrent, moderate to severe, distending (n = 9), pressure-like (n = 3) or pulsating (n = 3) pain within a strictly unilateral line-shaped area. The painful line is distributed from occipital or occipitocervical region to the ipsilateral eye (n = 5), forehead (n = 6) or parietal region (n = 1). The pain line has a trajecory similar to that of EF but no characteristics of moving. The headache duration would be ranged from five minutes to three days, but usually from half day to one day in most cases (n = 8). Six patients had the accompaniment of nausea with or without vomiting, and two patients had the accompaniment of ipsilateral dizziness. The attacks could be either spontaneous (n = 10) or triggered by noise, depression and resting after physical activity (n = 1), or by stress and staying up late (n = 1). The frequency of attacks was variable. The patients had well response to flunarizine, sodium valproate and amitriptyline but not to carbamazepine or oxcarbazepine. LH is different from EF, trigeminal autonomic cephalalgias (TACs) and cranial neuralgia, but it had couple of features similar to that of migraine. CONCLUSIONS: The clinical picture of LH might be a subtype of migraine, or represent a novel syndrome.

[Morphological study on early development of brain derived neurophic factor-positive neurons in the frontal lobe of human fetus].

OBJECTIVE: To investigate the growth and development of brain derived neurophic factor(BDNF)-positive neurons in the frontal lobe of human fetus. METHODS: The expression of the BDNF-positive neurons in the frontal lobe of human fetus in the 2(nd),3(rd),and 4(th) month of gestation were observed with the streptavidin-biotin-complex/immunoperoxidase(SABC)method. RESULTS: By the second month of gestation,BDNF-positive neurons were seen in the subventricular layer of the frontal lobe of cerebellum.By the third month of gestation,BDNF-positive neurons in the central layer were in various shapes,with big nucleus,less cytoplasm,and small processes.By the fourth month of gestation,BDNF-positive neurons in the central layer grew larger in size,cytoplasm increased,the BDNF-positive expression was enhanced with deeper dyeing,and the nerve fibers and particles were distributed between neurons;also,the BDNF-positive neurons were seen in the marginal layer of the frontal lobe of cerebrum. CONCLUSION: BDNF-positive neurons may participate in the early development of the frontal lobe of cerebrum of human fetus.

[Influence of leukoaraiosis on the cognition of patients with Parkinson disease].

OBJECTIVE: To explore the role of leukoaraiosis (LA) on the cognitive function in patients with Parkinson disease (PD). METHODS: The cohort for this study included 63 patients with PD, whom were divided into 3 groups according to cognitive status:with intact cognition (PD-IC, n = 23), with mild cognitive impairment (PD-MCI, n = 23) and with dementia (PDD, n = 17). All the patients were recruited from the Department of Neurology, First Affiliated Hospital, Anhui Medical University between September 2011 and July 2012. The cognitive functions were evaluated by mini-metal state examination (MMSE), the Cambridge cognitive examination-Chinese version (CAMCOG-C), clinical dementia rating (CDR), clock drawing task (CDT) and verbal fluency test, etc. Depression symptoms were assessed by the geriatric depression scale (GDS) while motor symptoms by the Unified Parkinson's Disease Rating Scale-motor (UPDRS-motor) and the Hoehn and Yahr scale (HY). All the patients underwent magnetic resonance imaging (MRI) with a 3.0-T system. LA was rated using the semiquantitative visual rating system proposed by scheltens et al. RESULTS: Both the PD-IC (2.43 ± 2.79) and PD-MCI (4.48 ± 4.33) groups showed significantly lower deep hyperintensities (DHs) scores than the PDD group (7.88 ± 6.69, P = 0.004 and 0.040, respectively), especially in frontal (1.09 ± 1.31; 1.83 ± 1.90; 3.24 ± 2.64, P < 0.05) and parietal areas (0.09 ± 0.29; 0.65 ± 1.03; 1.53 ± 2.32, P < 0.05). There were no significant differences in periventricular (1.57 ± 1.75; 2.52 ± 2.37; 3.24 ± 2.64, P > 0.05), basal ganglia (0.09 ± 0.42; 0.30 ± 0.77; 0.53 ± 1.33, P > 0.05) and infratentorial white matter hyperintensities scores (--; 0.13 ± 0.63; 0.18 ± 0.73, P > 0.05) among three groups. The DHs showed a significant correlation with age (P = 0.003), MMSE (P = 0.009), verbal fluency test (P = 0.009), orientation (P = 0.047) and executive function (P = 0.027) in CAMCOG-C. The multiple regression analysis showed that the MMSE scores were associated significantly with education (P < 0.001, ß = 0.600), DHs (P = 0.001, ß = -0.678) and HY (P = 0.035, ß = -0.480). DHs were the most significantly associated with MMSE scores. CONCLUSION: There was a significant correlation between DHs and multiple domain cognitive impairment in PD, especially in executive function. DHs, which were the most significantly variable associated with MMSE scores, may contribute to cognitive impairment in PD.

Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma.

UNLABELLED: In our previous study, osteopontin (OPN) was identified as one of the leading genes that promote the metastasis of hepatocellular carcinoma (HCC). However, the mechanism by which OPN promotes metastasis of HCC is not understood. In this study, RNA interference mediated by viral vectors-which could induce a long-lasting down-regulation in gene expression-was applied to analyze the role of OPN in metastasis of HCC. Three lentiviral vectors encoding microRNA against OPN, Lenti.OPNi-1, Lenti.OPNi-2, and Lenti.OPNi-3, were constructed and found to down-regulate the OPN level by 62%, 78%, and 95%, respectively, in HCCLM3 cells which had an overexpression of OPN and a higher metastatic potential. Consequently, both Lenti.OPNi-2 and Lenti.OPNi-3 induced a significant decrease in matrix metalloproteinase (MMP)-2 and urokinase plasminogen activator expression, and led to an obvious inhibition of both in vitro invasion and in vivo lung metastasis of HCCLM3 cells (P < 0.001). Moreover, Lenti.OPNi-3, rather than Lenti.OPNi-2, could also suppress in vitro proliferation and in vivo tumor growth of HCCLM3. Smaller detectable tumors were found in only 50% of mice after implantation of Lenti.OPNi-3-transfected HCCLM3 cells (341 +/- 502.6 mm(3) versus >3500 mm(3) in controls; P < 0.001). Lenti.OPNi-3, not Lenti.OPNi-2, significantly suppressed the MEK/ERK1/2 pathway in HCCLM3 cells. Recombinant OPN was found to induce translocation of p65 into the nucleus of HCC cells and activation of MMP-2 and MEK/ERK/1/2, which were suppressed by the nuclear factor kappaB (NF-kappaB) inhibitor pyrrolidine dithiocarbamate. CONCLUSION: OPN plays an important role in metastasis as well as tumor growth of HCC, in which different minimum threshold levels of OPN are needed. These effects may occur through activation of the mitogen-activated protein kinase and NF-kappaB pathways, and MMP-2. OPN could be a hopeful target for the control of HCC.

Identification of MSRA gene on chromosome 8p as a candidate metastasis suppressor for human hepatitis B virus-positive hepatocellular carcinoma.

BACKGROUND: The prognosis of patients with hepatocellular carcinoma (HCC) still remains very dismal, which is mainly due to metastasis. In our previous studies, we found that chromosome 8p deletions might contribute to metastasis of HCC. In this study, we aimed to identify the candidate metastatic suppressor gene on chromosome 8p. METHODS: Oligo-nucleotide microarrays which included 322 genes on human chromosome 8p were constructed to analyze the difference in gene expression profiles between HCC tissues with and without metastasis. The leading differentially expressed genes were identified and selected for further analysis by real-time PCR and Western blotting. Recombinant expression plasmid vectors for each target gene were constructed and transfected into HCC cells and its in vitro effects on proliferation and invasion of HCC cells were also investigated. RESULTS: Sixteen leading differentially expressed genes were identified from the HCC tissues with metastasis compared with those without metastasis (p < 0.01, q < 16 %). Among of the 10 significantly down-regulated genes in HCC with metastasis, methionine sulfoxide reductase A (MSRA) had the lowest p value and false discovery rate (FDR), and was considered as a potential candidate for metastasis suppressor gene. Real-time PCR and Western blotting confirmed that the mRNA and protein expression levels of MSRA were significantly decreased in HCC with metastasis compared with those without metastasis (p < 0.001), and MSRA mRNA level in HCCLM6 cells (with high metastatic potential) was also much lower than that of other HCC cell lines. Transfection of a recombinant expression plasmid vector and overexpression of MSRA gene could obviously inhibit cell colony formation (4.33 +/- 2.92 vs. 9.17 +/- 3.38, p = 0.008) and invasion (7.40 +/- 1.67 vs. 17.20 +/- 2.59, p= 0.0001) of HCCLM6 cell line. CONCLUSION: MSRA gene on chromosome 8p might possess metastasis suppressor activity in HCC.

[Knocking down osteopontin expression by specific siRNA reduces the in vitro invasiveness of human hepatocellular carcinoma cells].

OBJECTIVE: To study the effect of osteopontin (OPN) expression down-regulated by RNA interference (RNAi) on the invasiveness of hepatocelluar carcinoma cell line HCC-LM3. METHODS: HCC-LM3 cells were transfected with the chemically synthesized small interfering RNA (siRNA) formulated by lipofectamine 2000. Wild type HCC-LM3 and HCC-LM3 cells transfected with non-specific siRNA served as controls. Real-time PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The malignant phenotypes of transfected HCC-LM3 cells including cellular growth rate, colony formation and Matrigel invasion activities were analyzed. RESULTS: Sequence-specific siRNAs targeting OPN suppressed OPN RNA expression by 79% and also decreased OPN protein level by 81% in HCC-LM3 cells. The number of formed colonies and migrating numbers in vitro were decreased in HCC-LM3 cells transfected using sequence-specific siRNAs targeting OPN relative to controls (P < 0.05). CONCLUSION: This study demonstrated that specific siRNA is able to reduce OPN at both the mRNA and protein levels and significantly diminishes the invasiveness of hepatocellular carcinoma cells.

Expression of insulin-like growth factor binding protein-2 in gastric carcinoma and its relationship with cell proliferation.

AIM: To investigate the expression of insulin-like growth factor binding protein-2 (IGFBP-2) in gastric carcinoma and its clinical significance and to explore its relationship with cell proliferation. METHODS: Expressions of IGFBP-2 and Ki-67 in 118 cases of gastric carcinoma and 40 cases of normal gastric mucosa were detected by EnVision immunohistochemical technique. RESULTS: Expression of IGFBP-2 in gastric carcinoma was higher than that in normal gastric mucosa (P < 0.01). There was no difference between high- and low-grade gastric carcinoma (P > 0.05). Expression of IGFBP-2 in advanced gastric carcinoma was higher than that in early gastric carcinoma (P < 0.05). Expression of IGFBP-2 in gastric carcinoma with lymph node metastasis was higher than that without lymph node metastasis (P < 0.01). IGFBP-2 expression was a positively related to the clinical stage of gastric carcinoma (P < 0.01). There was a positive correlation between IGFBP-2 and Ki-67 (P < 0.05). CONCLUSION: IGFBP-2 may be involved in carcino-genesis and progression of gastric carcinoma by promoting cell proliferation.

KIAA0008 gene is associated with invasive phenotype of human hepatocellular carcinoma--a functional analysis.

PURPOSE: To investigate the function of the KIAA0008 gene, one of the leading genes in the signature associated with hepatocellular carcinoma (HCC) metastasis selected by cDNA microarray, and especially its possible roles in invasion and metastasis of hepatocellular carcinoma. METHODS: Expression levels of KIAA0008 in 27 primary tumors and 23 matched non-tumor liver tissues from HCC patients, and four HCC cell lines with different metastatic potentials were detected by semi-quantitative RT-PCR and real-time RT-PCR. Recombinant expression plasmid vectors of the KIAA0008 gene were constructed and transfected into HCC cells. The subcellular localization of the KIAA0008 gene product and in vitro effects of KIAA0008 overexpression on proliferation and invasion of HCC cell line were also investigated. RESULTS: Expression levels of KIAA0008 in HCC tissues were statistically higher than those of paired non-tumorous liver tissues (P < 0.001, paired Wilcoxon test), and in HCCs with high invasiveness these were statistically higher than those with low invasiveness (P = 0.002, Mann-Whitney test). In the four HCC cell lines with an identical genetic background and stepwise higher invasiveness potentials, its expression was consistent with their invasiveness potential. The KIAA0008 gene product was concentrated on the nucleus and cell membrane of HCC cells, without any distribution in the cytoplasm. Overexpression of KIAA0008 in the MHCC97L cell line resulted in increased cell proliferation, colony formation, and invasion. CONCLUSIONS: KIAA0008 expression is associated with invasiveness of HCC; overexpression of KIAA0008 leads to a more invasive phenotype of HCC cell lines.

[Expression of mucin MUC1 and MUC2 in colorectal carcinoma and their clinical significance].

BACKGROUND & OBJECTIVE: The up-regulation of mucin MUC1 expression or down-regulation of mucin MUC2 expression may be involved in carcinogenesis of some neoplasms, but there was few report about the relationship between mucin MUC1 and MUC2 expression and Chinese colorectal carcinoma. The aim of this study was to explore relationship between expression of mucin MUC1 and MUC2 in colorectal carcinoma and clinicopathological parameters and their clinical significance. METHODS: The expression of mucin MUC1 and MUC2 in 126 cases of colorectal carcinoma were detected by S-P immunohistochemical technique. RESULTS: The positive expression rates of mucin MUC1 and MUC2 in normal colorectal mucosa were 0 and 100%, respectively, but the positive expression rates of mucin MUC1 and MUC2 in 126 cases of colorectal carcinoma were 42.1% and 36.5%, respectively (P < 0.01); the expression of mucin MUC1 was positive correlated to the depth of invasion, lymph node metastasis, and Dukes staging (P < 0.05), but negative correlated to the degree of differentiation (P < 0.05); mucin MUC2 expression was related to invasion, lymph node metastasis, and Dukes staging (P < 0.05), but unrelated to differentiation (P > 0.05). CONCLUSIONS: The up-regulation of mucin MUC1 expression or down-regulation of mucin MUC2 expression may be involved in carcinogenesis and progression in colorectal carcinoma, and the detecting of mucin MUC1 and MUC2 may be predicting its prognosis in colorectal carcinoma.