Genome-wide identification of lncRNAs and mRNAs differentially expressed in non-functioning pituitary adenoma and construction of an lncRNA-mRNA co-expression network.

The involvement of long non-coding RNAs (lncRNAs) during tumorigenesis is a recent emerging theme. Yet no systematic evaluation of lncRNAs has been previously reported for non-functioning pituitary adenoma (NFPA), a fairly common type of intracranial tumor. Here, we report the first genome-wide expression profile for lncRNAs and mRNAs in NFPA, using formalin-fixed and paraffin-embedded tissue specimens. Using microarray analyses, we identified 113 lncRNAs and 80 mRNAs differentially expressed in NFPA; this list includes lncRNAs previously implicated in a variety of cancers. Using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) we further confirmed differential expression in NFPA for ten of the 113 lncRNAs. Using these ten doubly confirmed lncRNAs, we constructed an lncRNA-mRNA co-expression network comprising of 130 specific lncRNA-mRNA co-expression relationships. In addition, we conducted GO and KEGG analyses for the 80 mRNAs differentially expressed in NFPA. Our microarray and qRT-PCR analyses provided a working list of lncRNAs that may be functionally relevant to NFPA tumorigenesis. Our co-expression network in turn connected these largely uncharacterized lncRNAs to specific mRNAs, whose roles we further elucidated via GO and KEGG analyses, thus providing specific, testable hypotheses for the functions of these lncRNAs. Together, our study laid the foundation for future investigation of the specific function and mechanism by which lncRNAs are involved in NFPA tumorigenesis.

Human amniotic epithelial cells combined with silk fibroin scaffold in the repair of spinal cord injury.

Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithelial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the transplant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial cells combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.

Mobilization of Circulating Endothelial Progenitor Cells by dl-3-n-Butylphthalide in Acute Ischemic Stroke Patients.

OBJECTIVE: The research aim was to investigate the effects of dl-3-n-butylphthalide (NBP) on the level of circulating endothelial progenitor cells (EPCs) and clinical outcome in patients with acute ischemic stroke (AIS). MATERIALS AND METHODS: A total of 170 patients were included and randomly assigned to NBP group and control group. All patients were administrated a basic antiplatelet and lipid-lowering therapy. Among the patients, 86 received additional NBP administration for 30 days, whereas 84 received only basic therapy (the control). The level of circulating EPCs (marked with CD34(+)/CD133(+)/KDR(+)) was determined by flow cytometry at baseline and days 7, 14, and 30 after therapy. Impairment of neurological function was evaluated by the National Institutes of Health Stroke Scale (NIHSS) on days 7, 14, 30, and 90 after therapy. The association between the increased level of circulating EPCs and improvement of NIHSS score was evaluated by Pearson analysis. The clinical outcome was evaluated by modified Rankin Scale (mRS) on day 90. During the observation period, any adverse events related to drugs were reported. RESULTS: The levels of circulating EPCs on days 14 and 30 were significantly higher in the NBP group than in the control group. In contrast, NIHSS score was notably lower in NBP group on day 14, 30 and day 90. Pearson correlation analysis revealed a significant association between the increased level of EPCs and improvement of NIHSS score. Also, the mRS score in the NBP group was lower on day 90. Importantly, the reported adverse events in the 2 groups were comparable. CONCLUSION: NBP significantly increases the circulating level and improves clinical outcome in patients with AIS.

Expression of MMP14 in invasive pituitary adenomas: relationship to invasion and angiogenesis.

Pituitary adenomas (PAs) are noncancerous tumors, and about 35% of those reported to be invasive have been classified as "invasive pituitary adenomas (IPAs)". In clinical, operative complications, total resection failures, and high relapse rates result from invasive features during the therapeutic process. Invasive mechanism is a complex process, including metalloproteases, inhibitors and tumor microenvironment factors etc. Thus, studying invasive mechanism of PAs might contribute to understanding its biological behavior. In our research, three type tissue samples of human, pituitaries, PAs, IPAs, their mRNA expression of MMP1, MMP2, MMP9, MMP14 and MMP15 were measured using real-time PCR. MMP2 and MMP14 protein levels also were measured with immunohistochemistry in same samples. We confirmed that elevated matrix metalloproteinase-14 expression correlates with invasive characteristics of IPAs. To investigate molecular mechanism of how MMP14 contributes to invasiveness, an ATT20 cell was used in this study. After transient-transfection of the MMP14-shRNA expression vector into ATT20 cells, we observed that mRNA expression of PTTG, VEGF, and TGFß was significantly suppressed in interference groups. Meanwhile, ATT20 cells in high concentration TIMP-1 environment exhibit reduced PTTG, VEGF, and TGFß expression accompanied with the down-regulation of MMP14. Thus, we propose that MMP14 plays an important role in tumor invasion and angiogenesis and that a novel regulatory pathway for MMP14 may exist through VEGF and PTTG. In brief, MMP14 may be a target for therapeutic treatment.

Pituitary gene expression differs in D-galactose-induced cell senescence and steroid-induced prolactinomas.

In general, pituitary tumors are benign with low mitotic activity. Premature senescence has been considered to be a significant mechanism underlying this uniquely benign pituitary tumor. The present study aims to compare the expression of the associated proteins involved in premature senescence pathways among normal, aging and pituitary adenoma cells. We successfully induced the aging pituitary using continuous D­galactose (D­gal) injection as well as a prolactin­secreting pituitary tumor via diethylstilbestrol implants. Compared with normal pituitary cells, the aging pituitary tissues revealed increased expression of IL­6, C/EBPß, p53, p21 and p16 and decreased expression of pituitary tumor transforming gene. In contrast, the expression of IL­6, p21 and p16 was decreased in pituitary tumor cells compared with normal pituitary tissues. Taken together, multiple pathways including IL­6/C/EBPß, p53/p21 and p16 were activated in aging pituitary cells in response to D­gal treatment. However, all these pathways were immune to pituitary tumors treated by chronic estrogen. The findings and the involvement of cytokines in a highly prevalent natural disease model (pituitary adenomas) indicate a potential use of this pathway as a target for effective therapy for tumor silencing and prevention of adenoma progression towards malignancy.

Guard of delinquency? A role of microglia in inflammatory neurodegenerative diseases of the CNS.

Activation of microglia and inflammation-mediated neurotoxicity are believed to play an important role in the pathogenesis of several neurodegenerative disorders, including multiple sclerosis. Studies demonstrate complex functions of activated microglia that can lead to either beneficial or detrimental outcomes, depending on the form and the timing of activation. Combined with genetic and environmental factors, overactivation and dysregulation of microglia cause progressive neurotoxic consequences which involve a vicious cycle of neuron injury and unregulated neuroinflammation. Thus, modulation of microglial activation appears to be a promising new therapeutic target. While current therapies do attempt to block activation of microglia, they indiscriminately inhibit inflammation thus also curbing beneficial effects of inflammation and delaying recovery. Multiple signaling cascades, often cross-talking, are involved in every step of microglial activation. One of the key challenges is to understand the molecular mechanisms controlling cytokine expression and phagocytic activity, as well as cell-specific consequences of dysregulated cytokine expression. Further, a better understanding of how the integration of multiple cytokine signals influences the function or activity of individual microglia remains an important research objective to identify potential therapeutic targets for clinical intervention to promote repair.

Overweight, obesity and meningioma risk: a meta-analysis.

BACKGROUND AND OBJECTIVES: Studies of the association between excess body weight and risk of meningioma have produced inconsistent results. Therefore, a meta-analysis of published studies was performed to better assess the association between meningioma and excess body weight. METHODS: A literature search was conducted in the PubMed and EMBASE databases without any limitations. The reference lists of identified articles were also screened for additional studies. The summary relative risks (RRs) and 95% confidence intervals (CI) were calculated using fixed- or random-effects models. RESULTS: A total of 6 studies provided risk estimates for overweight or obesity. Overall, the combined RRs were 1.12 (95% CI = 0.98-1.28) for overweight and 1.45 (95% CI = 1.26-1.67) for obesity. After stratification by gender, no significant association was observed for obese men (RR = 1.30, 95% CI = 0.64-2.62), while significant association was detected for obese women (RR = 1.46, 95% CI = 1.26-1.69). No substantial differences emerged across strata of study design and geographic areas. CONCLUSION: The results of this meta-analysis suggest that obesity but not overweight is associated with an increased risk of meningioma. Due to the limited number of studies, further research is needed to confirm the association.

Alcohol consumption and risk of glioma: a meta-analysis of 19 observational studies.

The relationship between risk of glioma and alcohol consumption has been widely studied, but results have been conflicting. We therefore conducted a meta-analysis of observational studies to systematically assess the relationship between alcohol drinking and risk of glioma. Two electronic databases (PubMed and EMBASE) were searched from inception to 8 August 2013 to identify pertinent studies that linked alcohol drinking with glioma risk. We used a random-effects model to calculate the overall relative risk (RR) with corresponding 95% confidence intervals (CIs). Fifteen case-control and four cohort studies were identified for this analysis. The combined RR for total alcohol drinkers versus non-drinkers was 0.96 (95% CI: 0.89-1.04). In the subgroup analysis by geographic area, a significant association was observed in North American studies (RR = 0.78, 95% CI: 0.65-0.93), but not in European or Asian/Australian studies. In the subgroup analysis by study design, a borderline significant association emerged in population-based case-control studies (RR = 0.82, 95% CI: 0.68-0.99), but not in hospital-based case-control studies (RR = 1.00, 95% CI: 0.99-1.01) or cohort group (RR = 1.03, 95% CI: 0.88-1.20). Our results show no material association between alcohol consumption and risk of glioma existed. Further prospective evidences are needed to confirm this association.

Human amnion mesenchymal cells negative co-stimulatory molecules PD-L1 expression and its capacity of modulating microglial activation of CNS.

The objective of this study is to investigate the negative immunomodulatory capacity of human amniotic mesenchymal cells (AMSCs) and their possible intrinsic mechanism, by which we can confirm that they modulate microglial activation of central nervous system from multiple perspectives at the molecular level. The identification of the immune phenotype of AMSCs and microglial cells was executed by immunohistochemical methods and flow cytometry. Meanwhile, the influence and mechanism of amniotic mesenchymal cells in vitro on proliferation, cell cycle, and cytokine release of activated microglia (MI) would be detected by ELISA, ß-liquid scintillation counting method, and flow cytometry. Human amnion mesenchymal cells highly expressed negative co-stimulatory molecules PD-L1, while its ligand PD1 was expressed with high level by activated MI. When adding the PD-L1mAb to the mixed culture system composed of AMSCs and activated MI, the proliferation inhibitory effect and the cycle-blocking effect produced by the former on the latter would be partially reversed; at the same time, the impact of the latter cytokine secretion would be adjusted. As a conclusion, AMSCs play inhibitory effects on microglial activation, proliferation, and immune effects partially through the PD-L1-PD1 signaling pathways.

Personal hair dyes use and risk of glioma: a meta-analysis.

BACKGROUND AND OBJECTIVE: Use of hair dyes for glioma risk has been investigated in numerous epidemiological studies, but the evidence is inconsistent. Therefore, a meta-analysis was performed to estimate the association between hair dyes use and glioma risk. METHODS: We searched PubMed and EMBASE databases without any limitations, covering all papers published by the end of March 8, 2013. Cohort and case-control studies reporting relative risk estimates (RRs) with corresponding 95% confidence intervals (CIs) (or data to calculate them) on this issue were included. Random effects models were used to calculate the pooled RRs and corresponding 95% CIs. RESULTS: Four case-control and two cohort studies were included in this meta-analysis. The summary RRs and 95 % CIs for ever users of any hair dyes were 1.132 (0.887-1.446) for all studies, 1.291 (0.938-1.777) for case-control studies, and 0.903 (0.774-1.054) for cohort studies. In the subgroup analysis by geographic regions and sex, the similar results were detected. No significant associations were also observed among the studies which reported data involving permanent hair dye use and duration of any hair dye use. CONCLUSION: In summary, the results of our study demonstrated that hair dyes use is not associated with risk of glioma.

Exogenous and endogenous hormones in relation to glioma in women: a meta-analysis of 11 case-control studies.

BACKGROUND AND OBJECTIVE: Previous investigations of glioma risk in women have focused on oral contraceptive (OC), hormone replacement therapy (HRT), and reproductive factors. However, the results of published studies were inconclusive and inconsistent. Thus, a meta-analysis based on published case-control studies was performed to assess the role of exogenous and endogenous hormones factors in glioma risk. METHODS: The PubMed and EMBASE databases were searched without any restrictions on language or publication year. Reference lists from retrieved articles were also reviewed. We included case-control studies reporting relative risks (RRs) with corresponding 95% confidence intervals (CIs) (or data to calculate them) between oral contraceptive (OC) and hormone replacement therapy (HRT) use, reproductive factors and glioma. Random-effects models were used to calculate the summary risk estimates. RESULTS: Finally, 11 eligible studies with 4860 cases and 14,740 controls were identified. A lower risk of glioma was observed among women who were ever users of exogenous hormones (OC RR = 0.707, 95% CI = 0.604-0.828; HRT: RR = 0.683, 95% CI = 0.577-0.808) compared with never users. An increased glioma risk was associated with older age at menarche (RR = 1.401, 95% CI = 1.052-1.865). No association was observed for menopause status, parous status, age at menopause, or age at first birth and glioma risk. CONCLUSION: The results of our study support the hypothesis female sex hormones play a role in the development of glioma in women. Additional studies are warranted to validate the conclusion from this meta-analysis and clarity the underlying mechanisms.

Intraoperative microvascular Doppler monitoring in intracranial aneurysm surgery.

BACKGROUND: Surgical treatment of intracranial aneurysms is often compromised by incomplete exclusion of the aneurysm or stenosis of parent vessels. Intraoperative microvascular Doppler (IMD) is an attractive, noninvasive, and inexpensive tool. The present study aimed to evaluate the usefulness and reliability of IMD for guiding clip placement in aneurysm surgery. METHODS: A total of 92 patients with 101 intracranial aneurysms were included in the study. IMD with a 1.5-mm diameter, 20-MHz microprobe was used before and after clip application to confirm aneurysm obliteration and patency of parent vessels and branching arteries. IMD findings were verified postoperatively with digital subtraction angiography (DSA) or dual energy computed tomography angiography (DE-CTA). Ninety consecutive patients, harboring 108 aneurysms, who underwent surgery without IMD was considered as the control group. RESULTS: The microprobe detected all vessels of the Circle of Willis and their major branches. Clips were repositioned in 24 (23.8%) aneurysms on the basis of the IMD findings consistent with incomplete exclusion and/or stenosis. IMD identified persistent weak blood flow through the aneurismal sac of 11 of the 101 (10.9%) aneurysms requiring clip adjustment. Stenosis or occlusion of the parent or branching arteries as indicated by IMD necessitated immediate clip adjustment in 19 aneurysms (18.8%). The mean duration of the IMD procedure was 4.8 minutes. The frequency of clip adjustment (mean: 1.8 times per case) was associated with the size and location of the aneurysm. There were no complications related to the use of IMD, and postoperative angiograms confirmed complete aneurysm exclusion and parent vessel patency. About 8.3% (9/108) aneurysms were unexpectedly incompletely occluded, and 10.2% (11/108) aneurysms and parent vessel stenosis without IMD were detected by postoperative DSA or DE-CTA. IMD could reduce the rate of residual aneurysm and unanticipated vessel stenosis which demonstrated statistically significant advantages compared with aneurysm surgery without IMD. CONCLUSION: IMD is a safe, easily performed, reliable, and valuable tool that is suitable for routine use in intracranial surgery, especially in complicated, large, and giant aneurysms with wide neck or without neck.

Reproductive and exogenous hormone factors in relation to risk of meningioma in women: a meta-analysis.

BACKGROUND AND OBJECTIVE: A number of studies have focused on the association between oral contraceptive (OC), hormonal replacement therapy (HRT) and reproductive factors and meningioma risk, but the results were inconsistent. Thus, a meta-analysis was performed to obtain more precise estimates of risk. METHODS: We conducted a literature search using PubMed and EMBASE databases to July 2013, without any limitations. Random effects models were used to summarize results. RESULTS: Twelve case-control and six cohort studies were included in this meta-analysis. We found that an increased risk of meningioma was associated with HRT use(RR = 1.19, 95% CI = 1.01-1.40), postmenopausal women(RR = 1.32, 95% CI = 1.07-1.64) and parity(RR = 1.18, 95% CI = 1.00-1.40).No significant associations were observed for OC use (RR = 0.93, 95% CI = 0.83-1.03), age at menarche(RR = 1.06, 95% CI = 0.92-1.21), age at menopause(RR = 1.03, 95% CI = 0.81-1.30), or age at first birth(RR = 0.94, 95% CI = 0.80-1.10). CONCLUSION: In conclusion, the results of our study support the hypothesis that longer exposure to effect of female sex hormones may increase the risk of meningioma in women, yet additional studies are warranted to confirm our findings and identify the underlying biological mechanisms.

MiR-16-1 plays a role in reducing migration and invasion of glioma cells.

MicroRNAs (miRNAs) are novel small noncoding RNA molecules that regulate gene expression at the post transcriptional level. Compelling evidence shows that there are causative links between miRNAs deregulation and cancer development and progression. This study aims to explore the functions of miR-16-1 on proliferation, apoptosis, motility, and invasion of glioma cells. Quantitative real-time PCR (qRT-PCR) was performed to detect the expression of miR-16-1 in normal brain tissues and two glioma cell lines, including U251 and U87. CCK-8, Annexin V/FITC (fluorescein isothiocyanate), wound healing, and transwell assays were used to evaluate the functions of miR-16-1 that involves cell proliferation, apoptosis, motility, and invasion. In addition, we conducted qRT-PCR to examine mRNA expression levels of Zyxin, one of putative target genes of miR-16-1, in U251 glioma cells after transfecting with miR-16-1 mimics. As a result, miR-16-1 expression level was lower in U251 and U87 cells than normal brain tissues. After miR-16-1 was upregulated in U251 cells, cellular proliferation was notably attenuated but cell apoptosis was not significantly increased. Moreover, overexpression of miR-16-1 attenuated migration and invasion of glioma cells, and U251 cells transfected with miR-16-1 showed significantly lower endogenous mRNA levels of Zyxin than those transfected with nonspecific control miRNA or mock (P < 0.05). In summary, we demonstrated that miR-16-1 expression was markedly decreased in human glioma cell lines, and for the first time, described the roles of miR-16-1 in cellular proliferation, migration, and invasion abilities of high-invasive glioma cells, and suggested that Zyxin may be one of putative target genes of miR-16-1.

Elevated plasma endothelial microparticles in Alzheimer's disease.

BACKGROUND/AIMS: Endothelial microparticles (EMPs) in plasma are elevated in several vascular diseases. Alzheimer's disease (AD) is associated with microcirculatory injury, capillary blocking and disruption of the blood-brain barrier. We wanted to test the hypothesis that EMPs would be increased in AD patients and would correlate with a cognitive decline, and to determine if EMPs are released as a result of activation or apoptosis/necrosis in AD. METHODS: EMP levels in plasma of AD patients and controls were quantified by flow cytometry. EMP markers for apoptosis/necrosis [platelet/endothelial cell adhesion molecule-1 (PECAM-1)/CD31] and for activation (E-selectin/CD62e) were evaluated. The EMP CD62E/CD31 populations ratio of ≤1.0 was used to differentiate activation from apoptosis. RESULTS: Significantly higher CD31+/CD42- and CD62e+/CD42- counts were observed in the AD group relative to the controls (p < 0.05). There was no difference between the moderate- to-severe AD group and the mild AD group. Significant correlations were found between circulating EMP counts and Mini-Mental State Examination and AD Assessment cognition (ADAS-cog) score. Multivariate regression analysis demonstrated the persistence of significant correlations between ADAS-cog score and CD31+/CD42- EMPs. CONCLUSION: The (PECAM-1)/CD31 ratio demonstrated that EMPs were generated via apoptosis/necrosis and not by activation. Certain circulating EMP phenotypes may be associated with a cognitive decline of AD patients. EMP analysis shows a promising contribution to understanding vascular pathophysiology in AD.

Examination of the relationship between chromosome abnormality in pituitary adenomas and tumor invasiveness by normal karyotype analysis and interphase fluorescence staining.

To study the potential relationship between chromosome abnormality and tumor invasiveness in pituitary adenomas. To use conventional R-band cytogenetic karyotype analysis and interphase fluorescence in situ hybridization using centromeric probe of chromosomes 8, 9, and 11 to detect chromosome abnormality in 30 cases of pituitary adenoma. All chromosomes except chromosomes 4, 16, and Y show significant variation between invasive and noninvasive pituitary adenomas. Chromosomes 8 and 12 display some type of numeric alteration in all endocrine subtypes of pituitary adenoma. Numeric alterations in chromosomes 9, 11, and 19 are more frequently detected in invasive pituitary adenomas compared with noninvasive tumors. Numeric alterations in chromosomes are common in all endocrine subtype pituitary adenomas. Furthermore, chromosome numbers are significantly different in invasive and noninvasive pituitary adenomas. On the basis of our study and literature review, we conclude that while chromosomes 8 and 12 may play important roles in the occurrence of pituitary adenomas, chromosomes 9, 11, and 19 may be specifically associated with invasiveness.

Therapeutic effects of human amniotic epithelial cell transplantation on double-transgenic mice co-expressing APPswe and PS1ΔE9-deleted genes.

Human amniotic epithelial cells (HAECs), which exhibit characteristics of embryonic and pluripotent stem cells, could be utilized for cell therapy without legal or ethical problems. Double-transgenic (TG) mice (n=20) and wild-type (WT) mice (n=20) were randomly assigned to two groups, respectively. The transplantation group was treated with HAECs and the control group with PBS. A six-radial arm water maze was used to assess spatial memory. Immunofluorescence was utilized to track HAEC survival. Immunohistochemistry was used to determine octamer-binding protein 4 (oct-4) and nanog expression in the HAECs. High-performance liquid chromatography (HPLC) was used to measure acetylcholine levels in the hippocampus. The density of cholinergic neurons in the basal forebrain and nerve fibers in the hippocampus was measured following acetylcholinesterase staining. Results showed that transplanted HAECs survived for at least eight weeks and migrated to the third ventricle without immune rejection. Graft HAECs also expressed the specific stem cell markers oct-4 and nanog. Compared with the control group, HAEC transplantation significantly ameliorated spatial memory deficits in TG mice, as well as increased acetylcholine levels and the number of hippocampal cholinergic neurites. Intracerebroventricular HAEC transplantation improved spatial memory in double-TG mice, and results suggested that increased acetylcholine levels in the hippocampus, released by surviving cholinergic neurites, were responsible for this improvement.

Survival and regeneration of deep-freeze preserved autologous cranial bones after cranioplasty.

BACKGROUND: After decompressive craniectomy, a deep-freeze-preserved autologous cranial bone graft can be used for cranioplasty to avoid immunoreaction against an artificial patch material. Autologous cranial bone grafts not only have better physical properties, such as heat conduction, compared to artificial patch materials, but they also have the advantages of a lower medical cost and satisfactory physical flexibility. The discussion over (99)Tc(m)-MDP SPECT static cranial bone tomography in the diagnosis of survival and regeneration in deep-freeze preservation autologous cranial bones after cranioplasty is valuable. Objective. To investigate whether deep-freeze-preserved autologous cranial bone grafts could survive and regenerate after autologous reimplantation. METHODS: The method of cranial bone preservation involved removing the cranial graft and sealing it in a double-layer sterile plastic bag under sterile surgical conditions. On the day of the cranioplasty operation, the cranial bone graft was disinfected by immersing it in 3% povidone-iodine for 30 minutes. At short-term (2 weeks), medium-term (3 months), and long-term (12 months) postoperative follow-up visits, (99)Tc(m)-MDP SPECT static cranial bone tomography was used to examine the reimplanted cranial bone. Results. There were no postoperative infections or seromas in all 16 cases. Two weeks following cranial bone graft reimplantation, the SPECT tomography showed some radioactivity uptake in the reimplanted cranial bone graft, which was lower than that in the cranial bone on the healthy side. At 3 months and 12 months after the operation, the radioactivity uptake in the reimplanted cranial bone graft was the same as that in the cranial bone on the healthy side. X-ray films showed blurred sutures in the reimplanted cranial bone graft at 12 months after surgery. CONCLUSION: Reimplanted deep-freeze-preserved autologous cranial bone can survive in the short term and regenerate in the medium and long terms.

Human amniotic epithelial cells express specific markers of nerve cells and migrate along the nerve fibers in the corpus callosum.

Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes (microtubule-associated protein-2, glial fibrillary acidic protein and nestin) in human amniotic epithelial cells. The conditioned medium of human amniotic epithelial cells promoted the growth and proliferation of rat glial cells cultured in vitro, and this effect was dose-dependent. Human amniotic epithelial cells were further transplanted into the corpus striatum of healthy adult rats and the grafted cells could integrate with the host and migrate 1-2 mm along the nerve fibers in corpus callosum. Our experimental findings indicate that human amniotic epithelial cells may be a new kind of seed cells for use in neurograft.

Intracerebroventricular transplantation of human amniotic epithelial cells ameliorates spatial memory deficit in the doubly transgenic mice coexpressing APPswe and PS1ΔE9-deleted genes.

BACKGROUND: Human amniotic epithelial cells (HAECs), which have characteristics of both embryonic and pluripotent stem cells, are therefore a candidate in cell therapy without creating legal or ethical problems. In the present study, we aimed to investigate the effects of intracerebroventricular transplantation of HAECs on doubly transgenic mice of Alzheimer's disease (AD) coexpressing presenilin-1 (PS1) and mutant Sweden amyloid precursor protein (APPswe) genes. METHODS: The offspring mice genotypes were detected using PCR identification of APPswe and PS1 gene. The doubly transgenic (TG) mice (n = 20) and wild-type (WT) mice (n = 20) were randomly divided into two groups respectively: the transplantation group treated with HAECs and the control group with phosphate buffered saline. Six radial arm water maze test was used to assess the spatial memory in the TG and WT mice. Amyloid plaques and neurofibrillary tangles were analyzed using congo red and acid-silver methenamine staining respectively. Immunofluorescence cytochemistry was used to track the survival of HAECs. Immunohistochemistry was used to determine the expression of octamer-binding protein 4 (Oct-4) and Nanog in the HAECs. High performance liquid chromatography was used to measure acetylcholine in hippocampus. The density of cholinergic neurons in basal forebrain and nerve fibers in hippocampus was measured using acetylcholinesterase staining. RESULTS: Amyloid deposition occurred in hippocampus and frontal cortex in the double TG mice aged 8 months, but not in WT mice. The results also showed that transplanted HAECs can survive for at least 8 weeks and migrate to the third ventricle without immune rejection. The graft HAECs can also express the specific marker Oct-4 and Nanog of stem cell. Compared with the control group, transplantation of HAECs can not only significantly improve the spatial memory of the TG mice, but also increase acetylcholine concentration and the number of hippocampal cholinergic neurites. CONCLUSIONS: These results demonstrate that intracerebroventricular transplantation of HAECs can improve the spatial memory of the double TG mice. The higher content of acetylcholine in hippocampus released by more survived cholinergic neurites is one of the causes of this improvement.