Overexpression of a methyl-CpG-binding protein gene OsMBD707 leads to larger tiller angles and reduced photoperiod sensitivity in rice.

BACKGROUND: Methyl-CpG-binding domain (MBD) proteins play important roles in epigenetic gene regulation, and have diverse molecular, cellular, and biological functions in plants. MBD proteins have been functionally characterized in various plant species, including Arabidopsis, wheat, maize, and tomato. In rice, 17 sequences were bioinformatically predicted as putative MBD proteins. However, very little is known regarding the function of MBD proteins in rice. RESULTS: We explored the expression patterns of the rice OsMBD family genes and identified 13 OsMBDs with active expression in various rice tissues. We further characterized the function of a rice class I MBD protein OsMBD707, and demonstrated that OsMBD707 is constitutively expressed and localized in the nucleus. Transgenic rice overexpressing OsMBD707 displayed larger tiller angles and reduced photoperiod sensitivity-delayed flowering under short day (SD) and early flowering under long day (LD). RNA-seq analysis revealed that overexpression of OsMBD707 led to reduced photoperiod sensitivity in rice and to expression changes in flowering regulator genes in the Ehd1-Hd3a/RFT1 pathway. CONCLUSION: The results of this study suggested that OsMBD707 plays important roles in rice growth and development, and should lead to further studies on the functions of OsMBD proteins in growth, development, or other molecular, cellular, and biological processes in rice.

Bulked Segregant Analysis Coupled with Whole-Genome Sequencing (BSA-Seq) Mapping Identifies a Novel pi21 Haplotype Conferring Basal Resistance to Rice Blast Disease.

Basal or partial resistance has been considered race-non-specific and broad-spectrum. Therefore, the identification of genes or quantitative trait loci (QTLs) conferring basal resistance and germplasm containing them is of significance in breeding crops with durable resistance. In this study, we performed a bulked segregant analysis coupled with whole-genome sequencing (BSA-seq) to identify QTLs controlling basal resistance to blast disease in an F2 population derived from two rice varieties, 02428 and LiXinGeng (LXG), which differ significantly in basal resistance to rice blast. Four candidate QTLs, qBBR-4, qBBR-7, qBBR-8, and qBBR-11, were mapped on chromosomes 4, 7, 8, and 11, respectively. Allelic and genotypic association analyses identified a novel haplotype of the durable blast resistance gene pi21 carrying double deletions of 30 bp and 33 bp in 02428 (pi21-2428) as a candidate gene of qBBR-4. We further assessed haplotypes of Pi21 in 325 rice accessions, and identified 11 haplotypes among the accessions, of which eight were novel types. While the resistant pi21 gene was found only in japonica before, three Chinese indica varieties, ShuHui881, Yong4, and ZhengDa4Hao, were detected carrying the resistant pi21-2428 allele. The pi21-2428 allele and pi21-2428-containing rice germplasm, thus, provide valuable resources for breeding rice varieties, especially indica rice varieties, with durable resistance to blast disease. Our results also lay the foundation for further identification and functional characterization of the other three QTLs to better understand the molecular mechanisms underlying rice basal resistance to blast disease.

Evaluating the Rotation Correction of the Main Thoracic Curve in Severe Adolescent Idiopathic Scoliosis: Bending and Traction vs. Fulcrum - A Preliminary Report.

BACKGROUND Flexibility evaluation methods were only used to assess the changes of coronal Cobb angle in patients with adolescent idiopathic scoliosis (AIS). Little attention was attached to the vertebral rotation in these methods. MATERIAL AND METHODS 21 patients with severe adolescent idiopathic scoliosis were enrolled in this study. Coronal flexibility and rotation correction were compared on the supine bending, traction and fulcrum bending radiographs. The apical vertebral body rib ratio (AVB-R), and Perdriolle rotation angles were used to measure the rotation of the main thoracic curve. Statistical analysis was performed with one-way Analysis of Variance(ANOVA). Correlations between postoperative AVB-R and AVB-R in supine bending, traction and fulcrum bending radiographs were assessed utilizing the Linear Regression. RESULTS There were trends towards increased coronal flexibility in fulcrum bending versus traction versus supine bending, but there were no significant differences due to the limited sample size. And all were significantly lower than postoperative correction. The correction of AVB-R at traction and supine bending radiographs were significantly better than fulcrum bending, however, all were significantly lower than postoperative correction. Correction of Perdriolle rotation angle at traction radiograph was best among these methods. A univariant linear regression analysis showed a strong linear correlation between the postoperative AVB-R and the AVB-R in the traction radiograph. CONCLUSIONS As to patients with severe AIS, the coronal plane flexibility evaluated at the fulcrum bending radiograph is superior to that at the traction radiograph. This may be explained by the measurement errors induced by the better derotation capacity at the traction radiograph. Rotation correction evaluated at the traction radiograph proves better than the fulcrum bending radiographs, showing a linear correlation with the postoperative correction.

Whole-blood gene expression profiling in ankylosing spondylitis identifies novel candidate genes that may contribute to the inflammatory and tissue-destructive disease aspects.

We performed a comprehensive gene expression analysis to identify differentially expressed genes (DEGs) between AS (ankylosing spondylitis) and health controls. A total of 1454 DEGs were obtained, including 919 up-regulated genes and 535 down-regulated genes. There were 218 interactions and 224 pairs in the conPPI network. Topological analysis showed that 11 genes had a close relationship with AS. GO (gene ontology) functional enrichment analysis of the two modules showed that the DEGs in conPPI mainly participated in the biologic process of immune response. The KEGG pathway analysis showed that most DEGs in the two modules were enriched into cell receptor signaling pathway, natural killer cell mediated cytotoxicity and primary immunodeficiency. We hypothesized that these DEGs associated with immune response DEGs might provide basic for depth understanding of the AS development.

Comparison of the pedicle screws placement between electronic conductivity device and normal pedicle finder in posterior surgery of scoliosis.

STUDY DESIGN: Prospective randomized clinical trial. OBJECTIVE: To compare the accuracy and time using of pedicle screw placement between electronic conductivity device (ECD) and normal pedicle finder (NPF) in posterior surgery of scoliosis, through a randomized clinical trial. SUMMARY OF BACKGROUND DATA: Pedicle screw insertion for scoliosis correction can be associated with increased pedicle perforations. The malposition rates using various techniques in different region of the spine have been reported to occur with a frequency of 3.3%-43%. An ECD has been reported in spine surgeries, but its accuracy and surgical time comparing with NPF in the presence of scoliosis has not been reported. METHODS: The 42 patients of adolescent idiopathic scoliosis with average major Cobb angle of 55.3±7 degrees (range, 45-78 degrees), who received posterior correction surgeries using pedicle screws system only were divided into 2 groups by random: group NPF (22 patients); and group ECD (20 patients). NPF group had 332 screws and ECD group had 362 screws. The 2 groups were compared for accuracy of screw placement, time for screw insertion, and the number of times the C-arm had to be brought into the field. RESULTS: There were 47 (14.2%) pedicle perforation in the NPF group as compared with only 15 (4.1%) in the ECD group (P<0.001). Although in different region of the spine, screw accuracy showed discrepant statistical result, with upper (T1-T3), middle (T4-T7), and lower thoracic (T8-T10) comparison showing significant statistical difference (P=0.010, 0.001, and 0.041, respectively) and thoracolumbar (T11-L2) and lower lumbar (L3-L5) comparison showing no significant statistical difference (P=0.278 and 0.292, respectively). Average screw insertion time in the NPF group was 241±61 seconds compared with 204±33 seconds in the ECD group (P=0.009). The C-arm had to be moved into the operation field on an average of 1.59±0.67 times in the NPF group compared with 1.20±0.52 in the ECD group (P=0.040). CONCLUSIONS: ECD increases pedicle screw accuracy, especially in T1-T10, and reduces insertion time and radiation in posterior adolescent idiopathic scoliosis.

Expression and significance of Kruppel-like factor 6 gene in osteosarcoma.

PURPOSE: Osteosarcoma is primary malignant tumour of bone. Kruppel-like factor 6 (KLF6) is a tumor suppressor gene frequently inactivated in a number of human cancers and a ubiquitously expressed zinc-finger transcription factor. The present study aimed to first explore the relationship between the expression level of the KLF6 gene in osteosarcoma and the occurrence of bone tumours. METHODS: KLF6 mRNA and protein expression levels in osteosarcoma and normal bone tissue were assayed by real-time quantitative PCR and immunohistochemistry. KLF6 mRNA and protein expression levels in osteosarcoma cells and normal osteoblasts were detected by semi-quantitative reverse transcription PCR and Western blotting, respectively. RESULTS: Both the expression of KLF6 mRNA and protein in osteosarcoma cells and tissues were significantly lower than that in normal cells and tumour-adjacent tissues. CONCLUSIONS: KLF6 is a putative tumor suppressor gene involved in osteosarcoma which can be used as a new therapeutic target and an important marker for early diagnosis and postoperative monitoring.

Comparison between two pedicle screw augmentation instrumentations in adult degenerative scoliosis with osteoporosis.

BACKGROUND: The operative treatment of adult degenerative scoliosis combined with osteoporosis increase following the epidemiological development. Studies have confirmed that screws in osteoporotic spines have significant lower-screw strength with more frequent screw movements within the vertebra than normal spines. Screws augmented with polymethylmethacrylate (PMMA) or with autogenous bone can offer more powerful corrective force and significant advantages. METHODS: A retrospective analysis was conducted on 31 consecutive patients with degenerative lumbar scoliosis combined with osteoporosis who had surgery from December 2000. All had a minimum of 2-year follow-up. All patients had posterior approach surgery. 14 of them were fixed with pedicle screw by augmentation with polymethylmethacrylate (PMMA) and the other 17 patients with autogenous bone. Age, sex and whether smoking were similar between the two groups. Surgical time, blood loss, blood transfusion, medical cost, post surgery ICU time, hospital day, length of oral pain medicines taken, Pre-and postoperative Oswestry disability index questionnaire and surgical revision were documented and compared. Preoperative, postoperative and final follow up Cobb angle, sagittal lumbar curve, correction rate, and Follow up Cobb loss were also compared. RESULTS: No significant differences were found between the autogenous bone group and polymethylmethacrylate group with regards to all the targets above except for length of oral pain medicines taken and surgery cost. 2 patients were seen leakage during operation, but there is neither damage of nerve nor symptom after operation. No revision was needed. CONCLUSION: Both augmentation pedicle screw with polymethylmethacrylate (PMMA) and autogenous bone treating degenerative lumbar scoliosis combined with osteoporosis can achieve a good surgical result. Less oral pain medicines taken are the potential benefits of polymethylmethacrylate augmentation, but that is at the cost of more medical spending.

Factors affecting curve flexibility in skeletally immature and mature idiopathic scoliosis.

BACKGROUND: The determination of factors affecting curve flexibility is important in idiopathic scoliosis patients with regard to the Risser sign. The objective of this retrospective study was to identify factors affecting curve flexibility in patients with skeletally immature and mature idiopathic scoliosis. METHODS: The records of all patients with idiopathic scoliosis who received surgical treatment from July 2001 to August 2008 at our hospital were screened. The Risser sign was used to separate the patients into a skeletally mature group (Risser grade = 5) and skeletally immature group (Risser grade < 5). Data recorded and compared were flexibility (%), bending angle (°), apical vertebral rotation (°), Cobb angle (°), curve location, prior use of brace treatment, and number of vertebrae in the curve. RESULTS: The study cohort consisted of 217 patients (34 males, 183 females) in the Risser grade < 5 group and 124 (21 males, 103 females) in the Risser grade = 5 group. Multiple linear regression analysis revealed that the Cobb angle and the curve location significantly affected curve flexibility in the Risser grade < 5 group, whereas in the Risser grade = 5 group, Cobb angle and age significantly affected flexibility. CONCLUSIONS: Cobb angle and curve location influence main curve flexibility in skeletally immature adolescent idiopathic scoliosis, and Cobb angle and age influence curve flexibility in skeletally mature adult scoliosis. Measurement of these values may aid in the evaluation of treatment options and preoperative planning.

[The preliminary study of the efficiency of using cervical vertebral maturation of growth level of female adolescent idiopathic scoliosis].

OBJECTIVES: To investigate the reliability of cervical vertebral maturation (CVM) and to verify the possibility in the growth evaluation of female adolescent idiopathic scoliosis patients as a helpful supplementary to the Risser sign. METHODS: Coronal and lateral full-length spine X-ray film and left hand-wrist radiographs of 77 female adolescent patients with idiopathic scoliosis were selected from January 2010 to October 2010. The interval period between lateral length of the spine and left hand-wrist radiographs did not exceed 3 months. The CVM was assessed by a method developed by Baccetti and co-workers, whereas hand-wrist maturation was assessed by Fishman's method. The results were analyzed by Spearman correlation with patients Risser sign, chronological age, and menarche period. RESULTS: There were strong correlations between CVM and SMI or Risser sign (r = 0.862 and 0.762, P < 0.01). While in 26 patients whose Risser sign were 0-I, the correlation between CVM and SMI was more pronounced (r = 0.761, P < 0.01), compared with the correlation between Risser sign and SMI (r = 0.641, P < 0.01). CONCLUSIONS: CVM is a valid indicator of skeletal growth evaluation and can be used as a helpful supplementary to Risser sign.

Volume Navigation with Fusion of Real-Time Ultrasound and CT Images to Guide Posterolateral Transforaminal Puncture in Percutaneous Endoscopic Lumbar Discectomy.

BACKGROUND: The use of percutaneous endoscopic lumbar discectomy (PELD) is increasing in the treatment of lumbar disc herniations (LDH). Nerve and vessel injury may happen during the establishment of the transforaminal working channel. Also, there is usually exposure to intraop radiation when the C-arm is used to help establish the location of the transforaminal working channel. OBJECTIVE: To evaluate the accuracy and safety of the volume navigation technique for guiding transforaminal puncture in cadaver and clinical patient treatment. STUDY DESIGN: Cadaver experiment and comparative clinical research. SETTING: Changhai Hospital, the Second Military Medical University. METHOD: Volume navigation guided transforaminal puncture was performed in 15 cadavers. The registration error, time of overall puncture, ultrasound (US) observed distance between needle tip and target (DNT), and puncture error were recorded. Clinical research was performed in 63 patients who had undergone PELD. Comparative research was done between 2 groups: Those who had transforaminal puncture carried out under C-arm guidance (n = 30), and those patients whose transforaminal puncture was carried out under volume navigation guidance (n = 33). Puncture times and frequency of fluoroscopy were recorded. Both groups were evaluated with Oswestry Disability Index (ODI), and visual analog scale (VAS) before surgery at 1, 3, and 6 months, and 1 year post-surgery. RESULTS: In the cadaver experiment, mean registration error was 2.66 ± 1.10 mm; DNT 20.08 ± 1.32 mm; puncture error 2.91 ± 1.29 mm; overall time of puncture 22.10 ± 5.20 min. In the clinical patient research, puncture times and frequency of fluoroscopy were significantly lower in the volume navigation group compared with the C-arm group (P < 0.001). There were no significant differences between the 2 groups in ODI and VAS scores (P > 0.05) at different time points. LIMITATIONS: The correlation between the registration errors and the puncture errors requires further analysis. Also, due to the relatively small number of cases studied, additional cases need to be collected to obtain reliable results. CONCLUSION: The volume navigation technique can be used for PELD because it helps to guide percutaneous posterolateral transforaminal puncture accurately with reduced puncture times and intraop radiation. KEY WORDS: Lumbar disc herniation (LDH), ultrasound volume navigation (US VNav), percutaneous endoscopic lumbar discectomy (PELD), foramen, puncture.

Emodin enhances the demethylation by 5-Aza-CdR of pancreatic cancer cell tumor-suppressor genes P16, RASSF1A and ppENK.

5-Aza-2'-deoxycytidine (5-Aza-CdR) is currently acknowledged as a demethylation drug, and causes a certain degree of demethylation in a variety of cancer cells, including pancreatic cancer cells. Emodin, a traditional Chinese medicine (TCM), is an effective monomer extracted from rhubarb and has been reported to exhibit antitumor activity in different manners in pancreatic cancer. In the present study, we examined whether emodin caused demethylation and increased the demethylation of three tumor-suppressor genes P16, RASSF1A and ppENK with a high degree of methylation in pancreatic cancer when combined with 5-Aza-CdR. Our research showed that emodin inhibited the growth of pancreatic cancer Panc-1 cells in a dose- and time-dependent manner. Dot-blot results showed that emodin combined with 5-Aza-CdR significantly suppressed the expression of genome 5mC in PANC-1 cells. In order to verify the effect of methylation, methylation-specific PCR (MSP) and bisulfite genomic sequencing PCR (BSP) combined with TA were selected for the cloning and sequencing. Results of MSP and BSP confirmed that emodin caused faint demethylation, and 5-Aza-CdR had a certain degree of demethylation. When emodin was combined with 5-Aza-CdR, the demethylation was more significant. At the same time, fluorescent quantitative PCR and western blot analysis results confirmed that when emodin was combined with 5-Aza-CdR, the expression levels of P16, RASSF1A and ppENK were increased more significantly compared to either treatment alone. In contrast, the expression levels of DNA methyltransferase 1 (DNMT1) and DNMT3a were more significantly reduced with the combination treatment than the control or either agent alone, further proving that emodin in combination with 5-Aza-CdR enhanced the demethylation effect of 5-Aza-CdR by reducing the expression of methyltransferases. In conclusion, the present study confirmed that emodin in combination with 5-Aza-CdR enhanced the demethylation by 5-Aza-CdR of tumor-suppressor genes p16, RASSF1A and ppENK by reducing the expression of methyltransferases DNMT1 and DNMT3a.

[Effects of acrylonitrile in drinking water on monoamine neurotransmitters and its metabolites in male rat brains].

OBJECTIVES: To elucidate the possible involvement of monoamine neurotransmitters in the development of neurobehavioral damage produced by acrylonitrile in drinking water in male rat brains. METHODS: Totally 30 male SD rats were randomly divided into three groups, the control group (n = 10), low dosage group (n = 10), and high dosage group (n = 10), which were respectively administered 0 mg/L, 50 mg/L, 200 mg/L acrylonitrile (AN) in drinking water. The treatment was lasted for 12 weeks. Seven animals were randomly selected from each group for determination of monoamine neurotransmitters in striatum and cerebellum by high performance liquid chromatography with electrochemical detector and activities of monoamine oxidase in cortex. RESULTS: The contents of dopamine in the striatum of low and high dosage groups were decreased to (2.2 +/- 0.7) and (3.2 +/- 2.0) microg/g wet tissue, respectively, and compared with that of control group (9.0 +/- 4.2) microg/g wet tissue, the differences were statistically significant. There were no statistical differences among the contents of dopamine in the cerebellum of all rats, and the levels of 3,4-dihydroxyphenylacetic acid (DOPAC), the major metabolite of dopamine in the cerebellum were (186 +/- 41), (245 +/- 90) and (115 +/- 65) ng/g wet tissue in the control, low and high dosage groups, respectively and in low-dosage group they were significantly higher than those in other groups. There was dosage-dependently decreasing of the contents of serotonin of striatum in the control (249 +/- 34) ng/g wet tissue, low dosage (155 +/- 95) ng/g wet tissue and high dosage groups (128 +/- 101) ng/g wet tissue. CONCLUSION: This study underlines the importance of alterations in the monoamine neurotransmitters system as a possible causative mechanism behind the behavioural and functional changes produced by acrylonitrile.

[Construction of online medical parasites database].

We aim to construct a online medical parasites database from which information can be retrieved through key words, ASP program and SQL language. The connection between the web pages and the database was established to enable relevant web page searching and parasitological information retrieval on the basis of the database files. A method for online searching through the database is elaborated in this paper by introducing the construction of the medical parasites database.

Factors as predictors for thoracic and thoracolumbar/lumbar structural curves in adolescent idiopathic scoliosis.

BACKGROUND: Recent studies have demonstrated that the Lenke system is relatively efficient and consistent in classifying scoliosis curves. Basically, fusion should include the main curve and the structural minor curve. The criteria for defining the structural minor curve were established to help guide these decision-making process. The present study was designed to investigate predictors of the structural curve, and see whether it was possible to prevent the formation of the structural curve by interfering with influencing factors to decrease the fusion level. METHODS: Age, gender, Cobb angle, Perdriolle rotation, Risser sign and the number of vertebrae included in the curve, brace treatment, and curve location were recorded in 145 idiopathic scoliosis patients from July 2001 to January 2007. The patients were divided into two groups: structural and non-structural groups. Demographics and baseline characteristics were compared between the two groups as an initial screen. Logistic regression was used to analyze factors affecting the minor curve to become the structural curve. RESULTS: Compared with the non-structural group, the structural group had a higher Cobb angle ((51.34 ± 13.61)° vs. (34.20 ± 7.21)°, P < 0.001), bending angle ((33.94 ± 9.92)° vs. (8.46 ± 5.56)°, P < 0.001) and curve rotation ((23.25 ± 12.86)° vs. (14.21 ± 8.55)°, P < 0.001), and lower flexibility ((33.48 ± 12.53)% vs. (75.50 ± 15.52)%, P < 0.001). There was no significant difference in other parameters between the two groups. The results of the Logistic regression analysis showed that the Cobb angle (OR: 9.921, P < 0.001) and curve location (OR: 4.119, P = 0.016) were significant predictors of structural curve in adolescent idiopathic scoliosis. Every 10° change of Cobb angle increased the possibility of turning the minor curve into the structural curve by 10-fold. And thoracic curve showed, on the average, the possibility of becoming the structural curve about 4-fold more often than did the thoracolumbar/lumbar curve. CONCLUSIONS: Curve severity and curve location affect the minor curve's structural features in adolescent idiopathic scoliosis.

Using precisely controlled bidirectional orthopedic forces to assess flexibility in adolescent idiopathic scoliosis: comparisons between push-traction film, supine side bending, suspension, and fulcrum bending film.

STUDY DESIGN: A prospective study. OBJECTIVE: To validate the effectiveness of push-traction film (PTF) in assessment of curve flexibility in adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: There is no agreement among surgeons about the most advantageous method in flexibility evaluation of scoliosis. As all methods available provide the orthopedic force from one direction and use a single torque, it is difficult for them to achieve the postoperative correction; also they could not meet the needs for different types of curves. METHODS: Precisely controlled bidirectional (push and traction) orthopedic forces were applied for curve flexibility evaluation in 31 consecutive adolescent idiopathic scoliosis patients. The correction rate (CR) of postoperation, supine side-bending, suspension, and fulcrum bending radiographs were compared with PTF in instrumented main thoracic (MT) and thoracolumbar/lumbar curves. Correlation and linear regression analyses were also been done to find the best predictor among the four methods. RESULTS: In MT group, CR of PTF was significantly higher than that of side bending (P = 0.010) and suspension (P = 0.000) but not significantly different from that of fulcrum bending (P = 0.335). In TL/L group, CR of PTF was significantly higher than that of suspension (P = 0.000), but not significantly different from that of side bending (P = 0.681) and fulcrum bending (P = 0.382). There was no significant difference between CR of PTF and postoperation in both MT (P = 0.122) and TL/L (P = 0.068) groups. Correlation and linear regression analyses showed that PTF provided the highest correlation of the four methods, with the postoperative angle in both MT (r = 0.957) and MT/L group (r = 0.779). CONCLUSION: To our knowledge, this was the first report about using precisely controlled bidirectional correction forces for curve flexibility evaluation. Although it did not achieve the best CR among the four methods studied, correlation and regression analyses confirmed that PTF was a more stable and accurate method to predict flexibility. We believe that further exploration of a more rational push-traction force ratio would help to obtain a better flexibility.

Learning curve of computer-assisted navigation system in spine surgery.

BACKGROUND: Spine surgery using computer-assisted navigation (CAN) has been proven to result in low screw misplacement rates, low incidence of radiation exposure and excellent operative field viewing versus the conventional intraoperative image intensifier (CIII). However, as we know, few previous studies have described the learning curve of CAN in spine surgery. METHODS: We performed two consecutive case cohort studies on pedicel screw accuracy and operative time of two spine surgeons with different experience backgrounds, A and B, in one institution during the same period. Lumbar pedicel screw cortical perforation rate and operative time of the same kind of operation using CAN were analyzed and compared using CIII for the two surgeons at initial, 6 months and 12 months of CAN usage. RESULTS: CAN spine surgery had an overall lower cortical perforation rate and less mean operative time compared with CIII for both surgeon A and B cohorts when total cases of four years were included. It missed being statistically significant, with 3.3% versus 4.7% (P = 0.191) and 125.7 versus 132.3 minutes (P = 0.428) for surgeon A and 3.6% versus 6.4% (P = 0.058), and 183.2 versus 213.2 minutes (P = 0.070) for surgeon B. In an attempt to demonstrate the learning curve, the cases after 6 months of the CAN system in each surgeon's cohort were compared. The perforation rate decreased by 2.4% (P = 0.039) and 4.3% (P = 0.003) and the operative time was reduced by 31.8 minutes (P = 0.002) and 14.4 minutes (P = 0.026) for the CAN groups of surgeons A and B, respectively. When only the cases performed after 12 months using the CAN system were considered, the perforation rate decreased by 3.9% (P = 0.006) and 5.6% (P < 0.001) and the operative time was reduced by 20.9 minutes (P < 0.001) and 40.3 minutes (P < 0.001) for the CAN groups of surgeon A and B, respectively. CONCLUSIONS: In the long run, CAN spine surgery decreased the lumbar screw cortical perforation rate and operative time. The learning curve showed a sharp drop after 6 months of using CAN that plateaued after 12 months; which was demonstrated by both perforation rate and operative time data. Careful analysis of the data showed CAN is especially useful for less experienced surgeon to reduce perforation rate and intraoperative time, although further comparative studies are anticipated.

Preferential expression of chemokine receptor CXCR4 by highly malignant human gliomas and its association with poor patient survival.

OBJECTIVE: CXCR4 is implicated in the growth, metastasis, and angiogenesis of malignant tumors. We investigated the potential role of CXCR4 in human gliomas. METHODS: The expression of CXCR4 messenger ribonucleic acid and protein by human glioma cell lines was examined by reverse-transcriptase polymerase chain reaction and immunocytochemistry analysis. Tumor cell chemotaxis and production of vascular endothelial growth factor induced by the CXCR4 ligand SDF-1beta were measured. Xenograft models were used for evaluation of glioma cell tumorigenesis. CXCR4 expression by xenografted tumors and primary human glioma specimens were evaluated for CXCR4 protein expression. The relationship between CXCR4 expression and patient survival was analyzed. A synthetic lipoxygenase inhibitor, Nordy, was tested for its effects on glioma cell expression and function of CXCR4, as well as on glioma cell tumorigenicity. RESULTS: CXCR4 expression correlated directly with the degree of malignancy of the human glioma cell lines and primary tumors. Activation of CXCR4 induced tumor cell chemotaxis and increased production of vascular endothelial growth factor. Glioma cells expressing higher levels of CXCR4 formed more rapidly growing and lethal tumors in nude mice. Primary human glioma specimens expressing CXCR4 contained high-density microvessels. Patients with CXCR4-positive gliomas had poorer prognosis after surgery. The lipoxygenase inhibitor Nordy diminished CXCR4 expression by glioma cell lines in vitro and reduced their tumorigenicity in nude mice. CONCLUSION: The level of CXCR4 expression seems to correlate with the degree of malignancy of human gliomas and may contribute to their rapid growth.

Activation of chemokine receptor CXCR4 in malignant glioma cells promotes the production of vascular endothelial growth factor.

Numerous studies have showed that chemokine receptors, such as CXCR4, contribute to the growth and metastasis of a variety of malignant tumors. In this study, we investigated the role of CXCR4 in the production of angiogenic factor, vascular endothelial growth factor (VEGF), in various human glioma cells from astrocytic origin. The expression of CXCR4 mRNA and protein in three glioma cell lines, U87-MG, SHG-44, and CHG-5, was determined by RT-PCR and immunocytochemistry, respectively. The malignancies of three gliomas were evaluated by expression of glial fibrillary acidic protein and vimentin, the differentiation markers of astrocytic cells. The role of functional CXCR4 in tumor cell migration was studied with chemotaxis assay. Ca2+ mobilization and VEGF production were measured in the cells after stimulation with CXCR4 ligand, SDF1beta. The results showed that the levels of functional CXCR4 expression at both mRNA and protein levels by several human glioma cell lines were correlated with the degree of differentiation of the tumor cells. Activation of CXCR4 induced glioma cell chemotaxis and could trigger the increase of intracellular [Ca2+]i. Such an activation could result in the increased production of VEGF by the stimulated tumor cells. Our results suggest that CXCR4 may contribute to the high level of VEGF produced by malignant glioma cells and thus constitute a therapeutic target for antiangiogenesis strategy.