Objective To investigate the wake-promoting effect of vagus nerve stimulation (VNS) on coma rats after traumatic brain in-jury (TBI), and the related mechanism. Methods A total of 168 healthy Sprague-Dawley rats were randomly divided into blank group, TBI group, antagonist group and VNS group, 42 rats in each group. The latter three groups were established TBI model with impact, and the rats in coma at least 30 minutes were included. VNS group accepted VNS, the antagonist group were injected intralateroventricularly Orexin A receptor 1 (OXR1) antagonist SB334867, and TBI group accepted sham VNS. Their behaviors were observed to determine the level of con-sciousness six, twelve and 24 hours after intervention, while the expression ofγ-aminobutyric acid b1 receptor (GABAb1R) in prefrontal cortex was detected with immunohistochemistry and Western blotting. Results There were 42 rats in the blank group, 11 rats in TBI group, 13 rats in the antagonist group, and 28 rats in VNS group awakened finally. The expression of GABAb1R in prefrontal cortex ranged as TBI group, antagonist group, blank group and VNS group from more to less twelve and 24 hours after intervention under Western blotting (F>60.412, P<0.001), and it ranged as TBI group, antagonist group, VNS group and blank group under immunohistochemistry (H=15.121, P=0.002), with no significant difference among time points (H=3.028, P=0.220). Conclusion VNS can promote waking from coma in rats after TBI, which may relate with the decrease of GABAb1R in prefrontal cortex that induced by Orexin A.

Objective To investigate the feasibility of labeling mice spleen lymphocytes with superparamagnetic iron oxide(SPIO)and in vitro MR imaging of the labeled cells.Methods Spleen lymphocytes of 5 mice were isolated and then labeled with SPIO of 100,50,25,15,10,5 μg/ml,which was previously prepared with PLL.Prussian blue staining was performed to show the intracellular iron.Cell viability was compared among fresh,labeled and unlabeled cells.Different concentrations of mice spleen lymphocytes were screened using 3.0 T MR on T2WI,T2 * WI and SWI sequences in vitro.Cell viability was compared using independent-sample t test between groups.The MRI values among different groups were compared using one-way ANOVA.Results SPIO prepared with PLL could successfully label mice spleen lymphocytes,the optimum concentration of SPIO was 5 μg/ml.The Prussian blue staining showed intracellular blue spots and a labeling efficiency of(93.6 ± 2.1)％.Three groups of fresh,labeled and unlabeled cells showed a Trypan blue staining result of(94.8 ± 3.1)％,(88.7 ± 2.7)％,and(88.9 ±3.2)％,respectively; no statistically significant difference was found in cell viability between labeled and unlabeled lymphocytes(t =0.281,P ＞ 0.05); however,the cell viability of fresh cells were statistically significant higher than the labeled and unlabeled lymphocytes(t =8.125 and 7.253 respectively,P ＜0.05for all).Among the T2 WI,T2 * WI and SWI sequences under the same concentrations of cells,the SWI sequence was the most sensitive.Conclusions The mice spleen lymphocytes can be effectively labeled with SPIO with no impact on cell viability,and MR can be used to track these labeled cells in vitro.The SWI sequence is the most sensitive.

ObjectiveTo investigate the molecular pathogenesis of a pedigree of X-linked spondyloepiphyseal dysplasia atarda (SEDL) and to establish methods of gene diagnosis. Methods Clinical diagnosis was made based on height measurement, radiological examination and pedigree analysis. Peripheral blood samples of relevant family members were collected. After genomic DNA extraction, single strand conformation polymorphism (SSCP) followed with DNA sequencing was used to detect SEDL gene exons 36. Microsatellite marker DXS16 was selected for linkage analysis. Results The abnormal electrophoretic bands were detected in exon 4 of probands by PCR-SSCP. A c. 218C ＞ T mutation in exon 4 of SEDL gene was found in three probands, which resulted in a change in amino acid sequence S37L. The heterozygous exon 4 mutation was identified in three carriers, but not in healthy individuals, and no mutations were detect in exon 3, 5 and 6 of probands. Three unmarried young females (Ⅲ10, Ⅳ6 and Ⅳ7) were found to harbor the mutation by DNA sequencing analysis. ConclusionsA c. 218C ＞ T missense mutation in exon 4 of SEDL gene is the cause of molecular pathogenesis of the pedigree. SSCP and DNA sequencing can be used for prenatal gene diagnosis.

Objective To explore the relationships between the quantitative parameters for contrastenhanced ultrasonography(CEUS) in hepatocellular carcinoma (HCC) and arteries in neoangiogenesis.Methods One hundred and fifteen HCCs in CEUS were analyzed off-line with dynamic vascular curve of ImageArena and immunohistochemical stained in the tissue slices.Parameters including maximum of intensity(IMAX),rise time(RT),time to peak(TTP),mean trasit time corresponding to the center of gravity(mTT) ,rise slope(RS) and washout time(WT) were analyzed by statistics.Unpaired arteries(UAs)of 82 HCC cases were succesfully stained.Results The number of UAs had moderate correlation with RT (r = - 0.446),TTP (r = - 0.432) and RS (r = 0.431) (P ＜ 0.05),and it had mild correlation with IMAX (r = 0.303) ,WT (r = 0.285) (P ＜0.05).Conclusions Neoangiogenesis is a functional adaptation to altered vascular dynamics.Quantitative parameters of CEUS,reflecting hemodynamics of tumors,are correlated with UAs.

In order to improve the interfacial bonding strength of hydroxyapatite/polyurethane implanted material and dispersion of hydroxyapatite in the polyurethane matrix, we in the present study synthesized nano-hydroxyapatite/polyurethane composites by in situ polymerization. We then characterized and analyzed the fracture morphology, thermal stability, glass transition temperature and mechanical properties. We seeded MG63 cells on composites to evaluate the cytocompatibility of the composites. In situ polymerization could improve the interfacial bonding strength, ameliorate dispersion of hydroxyapatite in the properties of the composites. After adding 20 wt% hydroxyapatite into the polyurethane, the thermal stability was improved and the glass transition temperatures were increased. The tensile strength and maximum elongation were 6.83 MPa and 861.17%, respectively. Compared with those of pure polyurethane the tensile strength and maximum elongation increased by 236.45% and 143.30%, respectively. The composites were helpful for cell adhesion and proliferation in cultivation.
Biocompatible Materials ; chemistry ; Cell Adhesion ; Cell Line ; Durapatite ; chemistry ; Humans ; Polymerization ; Polyurethanes ; Tensile Strength ; Transition Temperature

OBJECTIVE To evaluate the anti-tumor activities of WX-127-07,a new microtubule-tar-geting agent invitroand probe its molecular mechanism. METHODS The well-known microtubule-targe-ting anti-tumor drugs taxol,vincristine and anti-gout drug colchicine were used as positive controls. The anti-proliferation activity was examined in five different cell lines after treatment with WX-127-07(0.3 -300 nmol·L-1 )for 72 h by SRB assay. The cell cycle arrest profile was assayed by flow cytometry. The multiparameters of cytotoxicity,cell morphology,apoptosis and different signaling pathways related to tumorigenesis and inflammation were analyzed using the high content analysis platform. Tubulin tryptic digestion and competition inhibition assay for colchicine or vinblastine site were used to confirm the bind-ing site in microtubules at a molecular level. RESULTS All the tested compounds obviously inhibited the growth of A549,HepG2,HeLa,HLF and HUVEC cells. The lC50 values of WX-127-07 were 4.47±0.05, 5.18±0.08,4.90±0.19,4.10±0.16 and(5.04±0.08)nmol·L-1 respectively,lower than those of colchicine〔the lC50 values were 21. 17 ± 1. 22,14. 19 ± 0. 53,43. 80 ± 1. 64,145. 89 ± 10. 97 and( 27. 67 ± 1.79)nmol·L-1 ,respectively〕and those of vincristine〔the lC50 values were 16.51±0.36,16.76±0.33, 27.80±2.75,43.80±1.48 and(9.15±0.78)nmol·L-1 ,respectively〕,but were similar to or lower than those of taxol〔the lC50 values were 10. 68 ± 0. 61,12. 86 ± 0. 25,4. 81 ± 0. 61,102. 07 ± 15. 17 and( 3. 04 ± 0.12)nmol·L-1 ,respectively〕. High content multi-parameter analysis revealed that WX-127-07 induced a concentration-dependent microtubular depolymerization(P=0.0075)with the same pattern as colchicine and vincristine,but at a lower concentration. Both WX-127-07 and positive drugs could induce cell cycle arrest in A549 cells,increase nuclear membrane permeability and early signs of apoptosis in HepG2 cells,but neither cancer related pathways nor inflammation related pathways were affected. Microtubular competition inhibition assay showed that WX-127-07 inhibited the binding of colchicine with tubulin(P =0.0259). Tryptic digestion of tubulin-WX-127-07 premixture showed a similar electrophoretic band to that of tubulin-colchicine premixture. CONCLUSION WX-127-07 is a novel microtubule-depolymerizing agent with anti-proliferation activity and acting on the colchicine binding site.

BACKGROUND:Early-onset scoliosis is a kind of disease that seriously affects the growth of children’s spine and development of cardiorespiratory function. The treatment of the disease has always been the focus of many clinical researchers.OBJECTIVE:To analyze the therapy for early-onset scoliosis and explore the spinal fusion, spinal non-fusion, conventional growth rod technology and magnetic control ed growth rod technology of early-onset scoliosis. METHODS:We retrieved PubMed, CENTRAL, EMbase, the ISI Web of Knowledge Databases, VIP, CNKI, CBM and Wanfang Database for related studies published from inception of the database to March 2016. The key words were“scoliosis, growing rod, complications”. The included 54 studies were analyzed and discussed. RESULTS AND CONCLUSION:For these children of early-onset scoliosis, we should not only maintain the correction of spine deformities, but also protect the ability of spine growth, keeping the normal cardiopulmonary function. In addition to conventional (non-surgical) treatment, there are surgical treatment (such as spinal fusion and growing rod technique) and magnetical y control ed growing rod, a new technology for the treatment of early-onset scoliosis. A comprehensive understanding of the effect of surgical treatment on the spine growth and cardiopulmonary function of children with early-onset scoliosis wil help to prevent the occurrence of related complications, so as to obtain a better therapeutic effect.

Objective To investigate the clinical application of pedicle screw placement technique for pelvic external fixation.Methods A retrospective review was made on 63 patients (37 males and 26 females;23-67 years of age) managed with anterior or combined anterior and posterior external fixation of pelvic fracture between February 2011 and May 2014.According the ways of screw insertion,the study was divided into two groups:observation group (screw was inserted using the pedicle screw placement technique,n =36) and control group (screw was inserted directly,n =27).Tile fracture classification was 11 patients with type B1,7 type B2,7 type B3,5 type C1,3 type C2 and 3 type C3 in observation group,and 9 patients with type B1,7 type B2,4 type B3,4 type C1,2 type C2 and 1 type C3 in control group.Operation time,rate of the penetration,X-ray fluoroscopy times,screw loosening rate,and postoperative complication rate were documented.Radiologic evaluation of the pelvis was detected with the Matta' s criteria.At the final follow-up,functional evaluation of the pelvis was evaluated with the score proposed by Cole et al.Results Period of follow-up was (14.7 ± 3.2)months in observation group and was (13.8 ± 3.1) months in control group.A total of 190 screws were placed in observation group and 138 screws in control group.Better results were found in observation group than in control group with respect to operation time [(18.8 ± 4.1) min vs.(22.6 ± 5.4) min],rate of the penetration (1.6％ vs.8.7％),X-ray fluoroscopy times (1.6 ± 0.8 vs.2.2 ± 0.9),and screw loosening rate (1.6％ vs.6.5％).There were no significant difference between observation and control groups in percentage of good to excellent reduction(89％ vs.85％) and function score [(35.6 ±3.0) points vs.(34.8 ± 3.9) points] (P ＞ 0.05).After operation,3 patients with lateral femoral cutaneous nerve paralysis and none with wouud infection were found in observation group;3 patients with lateral femoral cutaneous nerve paralysis and one patient with wound infection were found in control group.Conclusion Either the pedicle screw placement technique or conventional technique achieves satisfactory clinical effect,but the former owns better results in operation time,screw insertion accuracy,fluoroscopy frequency and rate of screw loosening,suggesting a fast,safe,and effective screw placement method in pelvic external fixation that should be widely applied.

Treatment of calcaneal fractures has always been the focus of many clinical researchers.The goals of traditional surgical treatment are not only to restore the integrity of calcaneal articular surface but also to reconstruct the anatomy of the calcaneus.More importantly,we need to reduce postoperative soft tissue swelling and incidence of postoperative complications.In recent years,scholars have reported satisfactory clinical efficacy and prognosis resulting from a sinus tarsal approach for treatment of calcaneal fractures.This paper reviews the latest research progress concerning the sinus tarsal approach for treatment of calcaneal fractures at home and abroad,intending to provide helpful information for the clinical surgeons.

OBJECTIVE: To analyze the clinical phenotype and genetic variant in a Chinese pedigree affected with benign familial neonatal convulsion (BFNC). METHODS: Clinical data and peripheral blood samples of the pedigree were obtained with informed consent. Whole exome sequencing (WES) was carried out for the proband. Candidate variants were verified by Sanger sequencing. RESULTS: The pedigree comprised 9 individuals, among whom 4 were affected, including 3 males and 1 female. All patients had developed seizures during the neonatal period, which had ceased in 4 to 6 months. One patient had recurrence in between 1 and 2 years old. Genetic testing has identified a novel nonsense c.810G>A (p.W270X) variant in exon 5 of the KCNQ2 gene, which has co-separated with the BFNC phenotype in the pedigree. CONCLUSION The patients from this pedigree have conformed to the diagnosis of BFNC with good prognosis, which was in keeping with previously reported cases. The heterozygous c.810G>A (p.W270X) nonsense variant of the KCNQ2 gene probably underlay the pathogenesis of BFNC in this pedigree, which has expanded the mutational spectrum of the disease.
Asians/genetics* ; Child, Preschool ; China ; Epilepsy, Benign Neonatal/genetics* ; Female ; Genetic Testing ; Humans ; Infant ; Male ; Mutation ; Pedigree