[Expression and characterizations of a novel full-length gene PKIbeta related to human glioma].

OBJECTIVE: To obtain differentially expressed genes related to human glioma using cDNA microarray and to characterization of one novel full-length gene. METHOD: Four samples of human glioma samples, 1 fetal brain tissue sample, and 2 normal brain tissue samples were used to extract the total RNA, and the mRNA was used to make probes. After hybridization and washing procedure, the products of hybridization were scanned using computer system. One gene, named 436F11 clone, was subsequently analyzed by Northern blotting, bioinformatics, and protein expression. RESULT: Fifteen differentially expressed new genes related to human glioma were obtained through four times of hybridization and scanning. Northern blotting confirmed that over-expression of 436F11 gene in the normal human brain tissue and low-expression in the human glioma tissues. The analysis of BLASTn and BLASTx showed that the clone of 436F11 was a novel full-length gene coding 78 amino acids of protein with a theoretical relative molecular weight of 8648 and an isoelectric point of 4.69 and that it was 60% identical to mouse PKIbeta amino acid, so it was called human PKIbeta gene. After it was transfected into Escherichia coli, higher-expressed protein of PKIbeta was obtained which yielded a major clear band on an SDS-PAGE gel after purification. The products obtained from amino acid sequencing and molecular weight detection were exactly the same as the products performed by bioinformatic analysis. CONCLUSION: cDNA microarray technology can be successfully applied to identify differentially expressed genes. PKIbeta may be a novel full-length gene related to human glioma and may provide a new way to gene therapy of glioma.

Cabergoline for the treatment of bromocriptine-resistant invasive giant prolactinomas.

PURPOSE: Few studies have specifically focused on the effect of cabergoline on invasive giant prolactinomas (IGPs) resistant to bromocriptine. This study aims to evaluate whether cabergoline could be used as an effective therapy for patients with bromocriptine-resistant IGPs. METHODS: This retrospective study included 15 patients with bromocriptine-resistant IGPs who received treatment at our department during 2007-2015. Cabergoline was administered in all 15 patients, with the dose adjusted depending on the prolactin response of each patient. Parameters for outcome assessment included prolactin level, pituitary function, tumor size, improvement of the clinical symptoms, visual field defects, drug tolerance, and disease recurrence. RESULTS: Cabergoline was effective for the treatment of the patients with bromocriptine -resistant IGPs as represented by normalization of the prolactin level in 12/15 patients; improvement or normalization of the pituitary function in 4/5 patients with hypopituitarism; significant reduction in tumor size in 14/15 patients; and relief of the clinical symptoms in 11/15 patients. All 15 patients showed good tolerance to cabergoline. In addition, no recurrence was observed during the mean follow-up period of 63.47 (range 30-145) months. CONCLUSIONS: Data of this retrospective study demonstrate that cabergoline is an excellent option for the treatment of patients with bromocriptine-resistant IGPs.

A novel full-length gene of human ribosomal protein L14.22 related to human glioma.

BACKGROUND: This study was undertaken to obtain differentially expressed genes related to human glioma by cDNA microarray and the characterization of a novel full-length gene. METHODS: Total RNA was extracted form human glioma and normal brain tissue, and mRNA was used as a probe. The results of hybridization procedure were scanned with the computer system. The gene named 507E08 cone was subsequently analyzed by northern blot, bioinformatic approach, and protein expression. RESULTS: Fifteen differentially expressed genes were obtained from human glioma by hybridization and scanning for four times. Northern blot analysis confirmed that the 507E08 clone was low expressed in human brain tissue and over expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that the 507E08 clone was a novel full-length gene, which codes 203 amino acid of protein and is called human ribosomal protein 14.22 gene. The nucleotide sequence had been submitted to the GenBank with the accession number of AF329277. After expression in E. coli., protein yielded a major band of apparent molecular mass 22 kDa on an SDS-PAGE gel. CONCLUSIONS: cDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human ribosomal protein 13.22 may be correlated with the development of human glioma.

Transplantation of human amniotic epithelial cells improves hindlimb function in rats with spinal cord injury.

BACKGROUND: Human amniotic epithelial cells (HAECs), which have several characteristics similar to stem cells, therefore could possibly be used in cell therapy without creating legal or ethical problems. In this study, we transplanted HEACs into the injured spinal cord of rats to investigate if the cells can improve the rats' hindlimb motor function. METHODS: HAECs were obtained from a piece of fresh amnion, labeled with Hoechst33342, and transplanted into the site of complete midthoracic spinal transections in adult rats. The rats (n = 21) were randomly divided into three groups: Sham-operation group (n = 7), cells-graft group (n = 7), and PBS group (n = 7). One rat of each group was killed for histological analysis at the second week after the transplantation. The other six rats of each group were killed for histological analysis after an 8-week behavioral testing. Hindlimb motor function was assessed by using the open-field BBB scoring system. Survival rate of the graft cells was observed at second and eighth weeks after the transplantation. We also detected the myelin sheath fibers around the lesions and the size of the axotomized red nucleus. A one-way ANOVA was used to compare the means among the groups. The significance level was set at P < 0.05. RESULTS: The graft HAECs survived for a long time (8 weeks) and integrated into the host spinal cord without immune rejection. Compared with the control group, HAECs can promote the regeneration and sprouting of the axons, improve the hindlimb motor function of the rats (BBB score: cells-graft group 9.0 +/- 0.89 vs PBS group 3.7 +/- 1.03, P < 0.01), and inhibit the atrophy of axotomized red nucleus [cells-graft group (526.47 +/- 148.42) microm(2) vs PBS group (473.69 +/- 164.73) microm(2), P < 0.01]. CONCLUSION: Transplantation of HAECs can improve the hindlimb motor function of rats with spinal cord injury.

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.

cDNA microarray in isolation of novel differentially expressed genes related to human glioma and clone of a novel full-length gene.

BACKGROUND: This investigation was undertaken to obtain differentially expressed genes related to human glioma using cDNA microarray and the characterization of one novel full-length gene. METHODS: Total RNA was extracted from human glioma tissues and normal brain tissues, and mRNA was used to make probes. After hybridization and washing, the results were scanned using a computer system. The gene named 681F05 clone was an expressed gene to human glioma through four-time hybridization and scanning. Subsequently northern blot analysis was performed by northern blot, 5'RACE and bioinformatics. RESULTS: Fifteen differentially expressed genes to human glioma were obtained through four-time hybridization and scanning. Northern blot analysis confirmed that 681F05 clone was low-expressed in human brain tissues and over-expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that 681F05 clone is two cDNA clones encoding two novel proteins that are highly identified to the cyclophilin isoform 10 of C. Elgans, respectively. Sequence analysis revealed the two cDNA clones are two different splicing variants of a novel cycophilin-like gene (PPIL3a and PPIL3b). CONCLUSIONS: cDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human PPIL3 may be correlated with the formation of human glioma.

Dopaminergic trophism after intrastriatal injection of lentivirus-transferred GDNF in Parkinson rat model.

To investigate the effects of lentivirus-mediated transfection of GDNF on Parkinson s disease (PD). The pNL-gdnf plasmid was constructed by replacing the LacZ-coding region present in pNL-lacZ/CMV. Vector particles involved a three-plasmid lentivirus expression system were co-transferred into 293T cells through calcium phosphate method. High-titer virus was collected from infected 293T cells and injected into lesion-side striatum of PD rats, and their apomorphine-induced rotations were assayed at day 14, 30 and 60, respectively. GDNF protein was detected by Western blot analysis, and the expression of lacZ and TH were detected by immunochemistry. Results showed that behavioral recovery gradually appeared after transplantation, and a significant reduction in the rotational response was observed at the 14th day. Meanwhile, gdnf expression maintained for at least 60 d, which had dopaminergic trophism to a certain degree, indicating that lentivirus-mediated transfection gdnf could effectively improve the clinical function of PD rats, which provide a potential attractive tool of gene therapy for PD.

Adipose tissue-derived stromal cells express neuronal phenotypes.

BACKGROUND: Adipose tissue-derived stromal cells (ADSCs) can be greatly expanded in vitro, and induced to differentiate into multiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic, and adipogenic cells. This study was designed to investigate the possibility of ADSCs differentiating into neurons. METHODS: Adipose tissue from rats was digested with collagenase, and adherent stromal cells were cultured. A medium containing a low concentration of fetal bovine serum was adopted to induce the cells to differentiate. ADSCs were identified by immunocytochemistry, and semi-quantitative RT-PCR was applied to detect mRNA expression of neurofilament 1 (NF1), nestin, and neuron-specific enolase (NSE). RESULTS: Nestin-positive cells were found occasionally among ADSCs. ADSCs were found to express NSE mRNA and nestin mRNA, but not NF1 mRNA. ADSCs could differentiate into neuron-like cells in a medium composed of a low concentration of fetal bovine serum, and these differentiated cells displayed complicated neuron-like morphologies. CONCLUSIONS: The data support the hypothesis that adipose tissue contains stem cells capable of differentiating into neurons. These stem cells can overcome their mesenchymal commitment, and may represent an alternative autologous stem cell source for CNS cell transplantation.

Culture of skin-derived precursors and their differentiation into neurons.

OBJECTIVE: To investigate the culture method of skin-derived precursors (SKPs) and to explore a new cell source for cell transplantation of central nervous system. METHODS: Cells from skins of juvenile and adult mice were isolated and cultured in serum-free medium. A mechanical method was chosen to passage these cells and they were identified by the immunocytochemistry assay. RESULTS: SKPs could be isolated from adult and neonatal skins. They could be maintained in vitro for long periods with stable proliferation, and expanded as undifferentiated cells in culture for more than 12 passages. About 50% of SKPs expressed nestin and majority of these cells expressed fibronectin when they were plated on polyornithine and laminin coated plates. About 5% cells showed neuronal differentiation and expressed neurofilament-M (NF-M) and NSE when SKPs were plated in serum-containing medium, and these cells could also differentiate into adipocytes and fibroblast-like cells. CONCLUSIONS: The data support the hypothesis that adult skin contains stem cells capable of differentiating into neurons, adipocytes, and fibroblast-like cells. They may represent an alternative autologous stem cell source for CNS cell transplantation.

The in vitro myelin formation in neurospheres of human neural stem cells.

OBJECTIVE: To explore the culture conditions of human neural stem cells and to investigate the ultrastructure of neurospheres. METHODS: The cells from the embryonic human cortices were mechanically dissociated. N2 medium was adapted to culture and expand the cells. The cells were identified by immunocytochemistry and EM was applied to examine the ultrastructure of neurospheres. RESULTS: The neural stem cells from human embryonic brains were successfully cultured and formed typical neurospheres in suspension, and most of the cells expressed vimentin, which was a marker for neural progenitor cells, and the cells could differentiate into neurons, astrocytes and oligodendrocytes. In vitro myelin formation in neurospheres were observed at an early stage of culture. CONCLUSIONS: Human neural stem cells can be cultured from embryonic brains, can form the typical neurospheres in suspension in vitro and have the ability of myelinating, and may be potential source for transplantation in treating myelin disorders.

[Adipose tissue-derived stromal cells as vector for gene therapy in central nervous system].

OBJECTIVE: To investigate exogenous gene expressing ability of adipose tissue-derived stromal cell (ADSCs) and cell distribution after they were transplanted into brains, and to get the genetically modified cells for autografting. METHODS: ADSCs were transfected by Ad5beta gal adenovirus containing a report gene, LacZ gene, then they were transplanted into the adult brain of rats, or ADSCs labeled by Hoechst33258 were transplanted into the adult brain of rats to investigate the migration and distribution of cells. RESULTS: ADSCs showed a good expression of LacZ with X-gal staining after transfecting and transplantation into adult brains, and they could incorporate into the host brain tissues and no disruption was observed. These cells showed good compatibility with the host brains. CONCLUSION: The results indicate that ADSCs could incorporate into host brains and express exogenous gene steadily when they were transplanted into adult brain tissues, no overproliferation and gliosis were identified, and ADSCs may be used as a therapeutic gene delivery vehicle in treating CNS disorders in humans.

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.

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.

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.

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.

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.