Background and purpose:The molecular regulation mechanism of the LCRG1 gnen is unclear;The study was designed to clarify the regulatory elements of LCRG1 gene in its 5'flanking region.Methods:Bioinformatics approaches were adopted and a putative promoter region was found in 5'flank upstream fragment of LCRG1 gene,5'flank upstream regulation fragment 1.776 kb(from-1 191 bp to +585 bp)of LCRG1 gene was amplified by PCR using genomic DNA as template,construct was obtained by cloning DNA fragments into pGL3 reporter vector.The construct was then introduced into COS7 cells by Lipofectemin method for transient expression of reporter gene,and luciferase activities was measured by luciferase assay.Results:The sequence of 5'flank upstream regulation fragment 1.776 kb(from-1 191 bp to +585 bp)of LCRG1 gene was successfully cloned and proved to be correct by DNA sequencing,the activity of pGL3-1776 was about 0.16-fold higher than that of pGL3-control cotransfection with PRL-TK and 35-fold higher than that of pGL3-basic cotransfection with PRL-TK.Conclusions:5'flank upstream regulation region 1.776 kb(from-1 191 bp to +585 bp)of LCRG1 was cloned successfully,the fragment presented promoter activity.

Objective To analyze the operation time,radiation exposure time and the screw placement accuracy of a newly-developed guiding chunnel-assisted percutaneous pedicle screw placement technique for thoracolumbar vertebral fractures not accompanied by nerve injury.Methods The clinical data of 35 patients with thoracolumbar vertebral fractures not accompanied by nerve injury,who were treated with newly-developed guiding chunnel-assisted percutaneous pedicle screw placement technique during the period from July 2010 to October 2012,were retrospectively analyzed.A total of 178 procedures of pedicle screw placement were performed in the 35 patients by the one and the same surgeon.The operation time and radiation exposure time of each pedicle screw placement procedure were recorded,and based on the findings of postoperative consecutive two CT scans of the operated vertebrae the screw placement accuracy was graded and evaluated.Results The technical success rate of screw placement was 100％.The mean time used for a single pedicle screw placement was (11.35±2.82) minutes,the average radiation exposure time was (8.06± 2.15) seconds.Screw placement accuracy of grade A was obtained in 156 screws (87.64％),grade B in 20 screws (11.24％),grade C in one screw (0.56％),and grade D in one screw (0.56％).Conclusion The newly-developed guiding chunnel-assisted percutaneous pedicle screw placement technique is very helpful in localizing the puncture point,in improving the screw placement accuracy,and in reducing both operation time and radiation exposure time.

Objective To study the effect of the continuous pulsing water ball in preventing infection in the multi-level posterior lumbar interbody fusion(PLIF).Methods From June 2016 to December 2016,thirty patients who were treated with multi-level posterior lumbar interbody fusion in our hospital were randomly divided into two groups.The 15 patients in the observation group were given continuous pulsing water ball,and the other 15 patients in the control group were given conventional dumping.The items including the time of intraoperative washing,the C-reactive protein(CRP),the time of suture clearing,postoperative fever cases and postoperative incision healing were oberved and compared between the two groups.Results The time of intraoperative washing was (2.13±0.15)min in the observation group and (5.59±1.24)min in the conrtrol group,and the difference was statistically significant (P<0.05).The postoperative CRP of 1st,3rd, and 5th day in the observation group was declining,whileas the control group was increasing.The time of suture clearing in the oberation group was 11.57 d, which was significantly less than 14.29 d the control group.The temperature of 3rd and 5th day after opreation in the obervation group was (37.1±2.26)℃ and (37.0±0.12)℃ respectively, which were significantly lower than (38.2±3.34)℃ and (37.5±0.25)℃ respectively the control group,and the difference was statistically significant(P<0.05).Conclusion Compared to the the conventional dumping,the continuous pulsing water ball has the some advantages in the multi-level posterior lumbar interbody fusion (PLIF),such as decreasing the washing time and reducing the local inflammatory reponse.

Objective:The regulatory mechanisms of LCRG1 gene expression is undear,this study was to investigate the transcription regulation of the promoter of human LCRG1 gene by transcription factors Sp1 and Egr-1.Methods:Analysis of putative binding sites of transcription factors in the promoter of human LCRG1 gene by online program MatInspector,Co-transfection with the expression plasmids of the known transcription factors such as Sp1、wtEgr-1、mtEgr-1with the LCRG1 reporter construct analyze potent transcription factor for enhancement of the promoter activity.Results:SP1 and Egr-1 transcription factor binding site sites were predicted in the region by bioinformatics analysis,mtEgr-1 can enhance the promoter activity.Conclusion:mtEgr-1 may be involved in the regulatory mechanisms of LCRG1 gene expression

BACKGROUND:Platelet-derived growth factor has the ability of wound repair, and relevant studies mainly focus on bone tissue repair. However, there are less studies about the effect of platelet-derived growth factors in skin wound healing. OBJECTIVE:To investigate the role of platelet-derived growth factor to promote wound healing by the regulation of bone marrow mesenchymal stem cels to the wound. METHODS:Bone marrow mesenchymal stem cels from rats were cultured. Immunofluorescence method was conducted to detect cel surface markers of CD34 and CD44 in bone marrow mesenchymal stem cels. Thirty healthy male SD rats were divided into five groups at random. Bone marrow mesenchymal stem cels labeled with PKH-26 were injected into the rat caudal vein in each group. The rats were anesthetized 1 week after injection. On the center of rat back, a 3-cm incision was made to establish a wound healing model. Different concentrations of platelet-derived growth factor were injectedvia multi-points on the skin wound after modeling, and the control group was treated with the same volume of normal saline. Skin wound tissues were taken for relevant parameter measurement at 14 days after injection. RESULTS AND CONCLUSION: Under the fluorescence microscope, platelet-derived growth factor could induce the migration and accumulation of bone marrow mesenchymal stem cels to the trauma in a dose-dependent manner and promote the wound healing. Masson staining showed that, with the concentration increase, platelet-derived growth factors could reduce inflammatory cel infiltration and increase the number of colagen fibers. Results from western blot assay showed that platelet-derived growth factor could inhibit the expression of matrix metaloproteinase-1, promote the expression of tissue inhibitor of matrix metaloproteinase 1 in the wound, and inhibit the colagen degradation, thereby promoting skin wound healing indirectly.

Purpose To explore the effect of EMS1-siRNA on the growth,invasion and migration of human gastric cancer cell line MGC803.Methods It used the Colony formation assay to determine the abilities of proliferation,and flow cytometry analysis to asses cell cycle distribution and apoptosis,transwell invasion and migration experiment to determine the ability of cell invasion and migration after knockdown the expression of EMS1 in MGC803.Results These results suggested that EMS1 gene down-regulated have no affect on cell cycle and cell apoptosis,but the ability of colony formation depressed and migration lowerd obviously (P ＜ 0.05).Conclusion The results shows EMS1 gene is related to proliferation and migration of tumor.

Laryngeal carcinoma related gene 1(LCRG1) is a candidate tumor suppressor gene of Laryngeal carcinoma. To further investigate its transcriptional regulation, the transcriptional start sites for LCRG1 gene have been identified by 5′ RACE (rapid amplification of cDNA ends) based on the bioinformation analysis of LCRG1. Then eleven luciferase expression vectors which contained potential human LCRG1 gene promoter were constructed. Luciferase reporter assay indicated that LCRG1 promoter region was mainly located in -169～+127 region nearby the major transcriptional start site. These results suggested that the region (-169～+127) includes an essential promoter for human LCRG1 gene transcription.

BACKGROUND:How to control the orderly formation of colage in skin repair and scarring process is worthy of attention. OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF) combined with insulin-like growth factor 1 (IGF-1) on the proliferation and colagen synthesis of rat bone marrow mesenchymal stem celsin vitro. METHODS:Rat bone marrow mesenchymal stem cels were isolated and cultured to induce adipogenic differentiation assessed by oil red O staining and osteogenic differentiation identified by alizarin red stainingin vitro. Passage 3 cels were cultured in the medium containing bFGF, IGF-1, combination of them or the control fluid, respectively. MTT assay was used to detect cel proliferation at 12, 24, 48, 72 and 96 hours of culture. The expression of type I colagen and type III colagen were detected by RT-PCR and western blot after 10 days of incubation. RESULTS AND CONCLUSION:Compared with the control group, bFGF or IGF-1 alone significantly promoted the proliferation of bone marrow mesenchymal stem cels, and inhibited the expression of type I colagen and type III colagen. After combined use of bFGF and IGF-1, the proliferation of bone marrow mesenchymal stem cels was improved more significantly, and the expression of type I colagen and type III colagen returned to normal levels. These findings indicate that the combination of IGF-1 and bFGF can promote proliferation of bone marrow mesenchymal stem cels and restrain the expression of type I colagen and type III colagen, which may be helpful for control and repair of scar formation during wound healing.

LCRG1(laryngeal carcinoma related gene1,LCRG1),a new candidate tumor suppressor gene of laryngeal carcinoma.However,it is known little about the possible regulatory mechanisms of LCRG1 gene expression.Restriction endonuclease digestion was used to obtain a set of the 5',or 3'deletion mutants from the region(-169～+127) of the LCRG1 gene.It has been found that the minimal promoter of the LCRG1 gene is mapped at the region from-169～-57.Linker scanning mutational analysis in the region(-169～+127) of the LCRG1 gene was used to identify the crucial cis-elements within the promoter region,The key cis-elements are within the region from-137～-122.SP1,E2F1/DP1,EKLF and ZF9 transcription factor binding site sites were predicted in the region by bioinformatics analysis.Co-transfection with each of a panel of the expression plasmids of the known transcription factors with the relevant reporter construct indicates Sp1 is potent transcription factor for enhancement of the promoter activity,SP1 can also up-regulate the endogenous expression of LCRG1 gene.Electrophoretic mobility shift assay(EMSA) was applied to verify that the key cis-elements of LCRG1 gene exist sequence of Sp1 binding sites.The findings,which showed that the key cis-elements within the region from 137～-122 play an important role in expression of the LCRG1 gene,provide a novel evidence for further study of the function of LCRG1 gene.

Objective Currently there is little study on the K-Rod pedicle fixation system in the treatment of far lateral lum-ber disc hernaition with multiple segment degeneration.The study was to compare the short-term efficay of K-Rod pedicle dynamic fixa-tion system and lumbar fusion for far lateral lumber disc herniation with multiple segment degeneration. Methods Retrospective a-nalysis were made on 56 patients with lumbar intervertebral disc herniation with multiple segment degeneration who received operation in General Hospital of Shenyang Military Region from January 2010 to October 2012.They were divided into K-Rod group (combined treatment of lumbar fusion and adjacent degenerative segment unfusion) and traditional fusion group according to different treatments. 28 patients in K-Rod group were treated with K-Rod pedicle dynamic fixation system and 28 cases in traditional fusion group were trea-ted with traditional lumbar fusion and internal fixation.Comparative analysis were made on the indexes before and after operation be-tween two groups, including Oswestry disability index, visual analogous scale (VAS) ,improvement rate of low back pain, postopera-tive lumbar spine X-ray, operation time, intraoperative blood loss and postoperative average length of stay. Results 12 to 24 months follow-up were achieved on 56 cases.The Oswestry function score and visual analog scale for both groups were improved significantly at 12 months after operation, and the difference was of statistical significance (P<0.05).Oswestry function scores and vis-ual analog scales between two groups were of no significant difference either before operation or after operation (P>0.05), while the average operation time ([119.6 ±14.2] min vs [155.5 ±17.5]min), the average blood loss ([235.5 ±32.5] mL vs [367.5 ±29. 5] mL), the average length of stay (ALOS) ([10.5 ±2.2] vs [14.2 ±2.6]d) between K-Rod group and traditional fusion group were of significant difference(P<0.05). Conclusion In the treatment of far lateral lumber disc hernaition with multiple segment degeneration, the K-Rod pedicle dynamic fixation system atrributes to less intraoperative blood loss, shorter operation time and little postoperative effect on lumbar spine mobility.