Dynamic Expression Profiles of Marker Genes in Osteogenic Differentiation of Human Bone Marrow-derived Mesenchymal Stem Cells

Objective To observe the expression profiles of osteoblast-related genes in human mesenchymal stem cells(MSCs) derived from bone marrow during osteogenic differentiation. Methods MSCs were induced to differentiate with MSC osteogenic differentiation medium for 7, 14, 21 and 28 days respectively. Alizarin Red staining was used to detect matrix mineralization. Expression of osteoblast-related genes, including osteocalcin, osteopontin, Runt-related transcription factor 2(Runx2), alkaline phosphatase and collagen type Ⅰ, was assessed with quantitative reverse transcription-polymerase chain reaction. Results On day 14 after induction of differentiation, cells were stained positively with Alizarin Red. The expression levels of these genes exhibited an upward trend as induction time was prolonged. Exposure to osteogenic differentiation medium less than 21 days did not significantly induce osteocalcin expression; osteocalcin expression levels in the differentiated cells induced for 21 and 28 days were 1.63 and 2.46 times as high as the undifferentiated cells respectively(all P<0.05). Stimulation with MSC osteogenic differentiation medium over 14 days significantly enhanced bone marrow-derived MSCs to express osteopontin and Runx2 genes(all P<0.05). Osteogenic differentiation medium could significantly induce the expressions of alkaline phosphatase and collagen type Ⅰgenes(all P<0.05). Their expressions reached the peak levels on day 21, which were increased more than 4- and 3-fold respectively. Conclusion Human bone marrow-derived MSCs could exhibit the sequential expression pattern of osteoblast marker genes during osteogenic differentiation in vitro.

Reliability of a Novel Cobb Protractor for Measuring the Cobb Angle of Radiograph in Scoliosis

Objective To introduce a novel Cobb protractor and assess its reliability and rapidity for measuring Cobb angle in scoliosis patients. Methods The novel Cobb protractor had two endplate markers. A measurement was performed just to align the two markers to each endplate of the curve. The Cobb angle on the posteroanterior radiographs of 24 patients clinically diagnosed with adolescent idiopathic scoliosis was measured by three orthopedic surgeons with both standard Cobb method and the new technique, and the time of measurement was recorded. Intraclass correlation coefficients(ICCs) were calculated to assess the reliability of the new method. Results The time for a measurement with the new tool was approximately 10 seconds less than the time that used to finish a measurement with the standard method(P<0.05). The overall mean Cobb angle for the major curve of the 24 patients was 47.8°. The mean overall intraobserver and interobserver ICC was 0.971 and 0.971 for the Cobb method group, while the overall intraobserver ICC and the interobserver was 0.985 and 0.979 for the new tool group. Conclusions The novel Cobb protractor could perform quick measurement and measure almost all forms of radiographs. The Cobb protractor might be an ideal instrument to measure the Cobb angle.

Phacoemulsification versus combined phacotrabeculectomy in the treatment of primary angle-closure glaucoma with cataract: a Meta-analysis

AIM： To compare the efficacy and safety of phacoemulsification（Phaco） against combined phacotrabeculectomy（Phacotrabe） in primary angle-closure glaucoma（PACG） with coexisting cataract.·METHODS： By searching electronically the Pub Med,EMBASE, Scientific Citation Index and Cochrane Library published up from inception to January 2014, all randomized controlled trials that matched the predefined criteria were included. The quality of included trials was evaluated according to the guidelines developed by the cochrane collaboration. And the outcomes estimating efficacy and safety of two different surgical treatments were measured and synthesised by Rev Man 5.0.· RESULTS： Five randomized controlled trials were selected and included in Meta-analysis with a total of468 patients（468 eyes） with both PACG and cataract. We found that Phacotrabe had a greater intraocular pressure（IOP） lowing effect [preoperative IOP： weighted mean difference（WMD）=0.58, 95% confidence intervals（95% CI,-0.53 to 1.69）, P =0.31; postoperative IOP： WMD =1.37,95% CI（0.45 to 2.28）, P =0.003], a lower number of antiglaucoma medications [risk ratio（RR）=0.05, 95% CI（0.02 to 0.18）, P 〈0.00001] needed postoperatively and less serious damage of optic nerve [RR =0.48, 95% CI（0.21 to 1.07）, P =0.07], but a higher risk of complications[odds ratio（OR）=0.04, 95% CI（0.01 to 0.16）, P 〈0.00001]compared with Phaco. The rest studies indicated that there had no significantly difference between the two surgical methods for postoperative best-corrected visual acuity（BCVA） [WMD =-0.05, 95% CI（-0.14 to 0.05）, P =0.32] and loss of visual field [RR=1.06, 95% CI（0.61 to1.83）, P =0.83].·CONCLUSION： Phaco alone compared with Phacotrabe had a better effect in IOP reduction, whereas the securitydecline. Considering the number of sample size, our results remains to be further studied.

Plasma levels of receptor interacting protein kinase-3 correlated with coronary artery disease

Background: Necroptosis plays an important role in human atherosclerosis and atheroma development. Since receptor interacting protein kinase-3 (RIP3) acts as a key mediator of necroptosis, this study aimed to explore its relationship between plasma RIP3 levels and coronary artery disease (CAD) and discover a potential new biomarker for screening CAD subtypes and severity. Methods: A total of 318 patients with CAD who had coronary angiography and 166 controls in Peking Union Medical College Hospital from September 2017 to January 2018 were enrolled in this study. Patients with CAD were divided into three subgroups: patients with stable coronary artery disease (SCAD), patients with unstable angina (UA), and patients with myocardial infarction (MI). The severity of atherosclerosis was determined by Gensini score (GSS). Logistic regression was used to determine the relationship between plasma RIP3 levels and CAD. The correlation between plasma RIP3 and GSS was calculated using multiple linear regression models. Results: Overall, plasma RIP3 levels were significantly higher than serum RIP3 levels. Plasma RIP3 levels in patients with CAD were significantly higher than those in controls. Plasma RIP3 levels were strongly associated with CAD (odds ratio: 6.00, 95% confidence interval 3.04–11.81;P < 0.001). Plasma RIP3 levels increased linearly from controls to patients with SCAD, then patients with UA, and finally to patients with MI. We found a significantly positive correlation between proportion of cases of acute coronary syndrome in subjects and their plasma RIP3 level quartile. Plasma RIP3 levels were also associated with GSS (B 0.027;standard error 0.012;P < 0.05). Conclusions: Plasma RIP3 levels were independently associated with CAD. Plasma RIP3 levels could potentially supplement clinical assessment to screen CAD and determine CAD severity.

Recent Advances in Technique and Clinical Outcomes of Minimally Invasive Spine Surgery in Adult Scoliosis

Objective： Conventional open spinal surgery of adult scoliosis can be performed from anterior, posterior, or combined approach. Minimally invasive spine surgery （MISS） was developed for the purpose of reducing the undesirable effects and complications. This review aimed to make a brief summary of recent studies of the approach and clinical outcomes of MISS in adult scoliosis. Data Sources： We conducted a systematic search from PubMed, Medline, EMBASE, and other literature databases to collect reports of surgical methods and clinical outcomes of MISS in treatment of adult scoliosis. Those reports were published up to March 2017 with the following key terms： ＆quot;minimally invasive,＆quot; ＆quot;spine,＆quot; ＆quot;surgery,＆quot; and ＆quot;scoliosis.＆quot;Study Selection： The inclusion criteria of the articles were as followings： diagnosed with adult degenerative scoliosis （DS） or adult idiopathic scoliosis; underwent MISS or open surgery; with follow-up data. The articles involving patients with congenital scoliosis or unknown type were excluded and those without any follow-up data were also excluded from the study. The initial search yielded 233 articles. After title and abstract extraction, 29 English articles were selected for full-text review. Of those, 20 studies with 831 patients diagnosed with adult DS or adult idiopathic scoliosis were reviewed. Seventeen were retrospective studies, and three were prospective studies. Results： The surgical technique reported in these articles was direct or extreme lateral interbody fusion, axial lumbar interbody fusion, and transforaminal lumbar interbody fusion. Among the clinical outcomes of these studies, the operated levels was 3–7, operative time was 2.3–8.5 h. Both the Cobb angle of coronal major curve and evaluation of Oswestry Disability Index and Visual Analog Scale decreased after surgery. There were 323 complications reported in the 831 （38.9%） patients, including 150 （18.1%） motor or sensory deficits, and 111 （13.4%） implant-related complications. Conclusions： MISS can provide good radiological and self-evaluation improvement in treatment of adult scoliosis. More prospective studies will be needed before it is widely used.

Effect of Resveratrol on Preventing Steroid-induced Osteonecrosis in a Rabbit Model

Background： Prevention of osteonecrosis （ON） has seldom been addressed. The purpose of this study was to evaluate the effect of resveratrol on preventing steroid-induced ON in rabbits. Methods： Seventy-two rabbits were divided into four groups： （l） NEC （ON） group： thirty rabbits were treated with lipopolysaccharide （LPS） once, then with methylprednisolone （MPS） daily for 3 days; （2） PRE （prevention） group： thirty rabbits were given one dose of LPS, then MPS daily for 3 days, and resveratrol on day 0 and daily for 2 weeks： （3） RES （resveratrol） group： six rabbits were given resveratrol for 2 weeks but without LPS/MPS; （4） CON （control） group： six rabbits were given alcohol for 2 weeks but without LPS/MPS. Levels of plasma tissue-type plasminogen activator （t-PAL plasminogen activator inhibitor 1 （PAl-1）, thrombomodulin （TM）, vascular endothelial growth factor （VEGF）, maximum enhancement （ME） by magnetic resonance imaging, and ON incidence were evaluated. Results： The PRE group had a lower ON incidence than the NEC group, but with no significant differences at 2 weeks and 12 weeks. Tile RES and CON groups did not develop ON. TM and VEGF were significantly higher in the NEC group compared with the PRE group at weeks 1, 2, and 4 （TM： 1 week, P= 0.029; 2 weeks, P= 0.005; and 4 weeks, P = 0.047; VEGF： 1 week, P= 0.039; 2 weeks, P= 0.021; 4 weeks. P = 0.014） but the difference disappeared at 12 weeks. The levels of t-PA and PAl-I were not significantly different between the NEC and PRE groups. The TM, t-PA, PAl-l, and VEGF concentrations in the RES and CON groups did not change over time. Compared to the baseline, ME in the NEC group decreased significantly （P = 0.025） at week 1, increased significantly （P = 0.021） at week 2, and was decreased at week 12. The variance was insignificant in the PRE group. Conclusions： Resveratrol may improve blood supply to bone in a rabbit model of ON of the femoral head via anti-inflammatory effects to protect the vascular endothelium and reduce thrombosis.

Lumbar Scoliosis Induction in Juvenile Dogs by Three？dimensional Modulation of Spinal Growth Using Nickel？Titanium Coil Springs

Background： Current treatments for scoliosis have some defects and complications. To study spinal deformities and test novel scoliosis treatments, many animal models of scoliosis have been developed. These models applied a single load to the spine and could not precisely modulate the spinal growth in different dimensions. In this study, we applied posterior tethering in various directions with the application of nickel-titanium （NT） coil springs in dog＇s spine to modulate spinal growth in the coronal, sagittal, and transverse planes and create a scoliosis model possess curves that mimic adolescent idiopathic scoliosis （AIS） three dimensionally. Methods： Scoliosis was surgically induced in eight 8-week-old female dogs （weight： 1.95–2.30 kg） using bone screws and NT coil springs. The deformity was induced through the placement of posterior NT coil springs that tethered the spine by bone screw fixation. All dogs were monitored with serial radiographs to document changes in deformities. Results： All experimental animals developed scoliotic curves convex to the left in the lumbar segment. The mean coronal Cobb angle was 18.0° immediately postoperatively and 54.5° at 22 weeks. The mean lordosis increased from 6.2° postoperatively to 35.0° at final follow-up. Apical axial rotation increased from 4.5° postoperatively to 31.2° at 22 weeks. Conclusions： With the application of NT springs in dogs that allowed posterior tethering in various directions, lumbar spinal deformity was achieved in three planes： coronal, sagittal, and transverse planes. Notably, the lumbar spine in surgically treated dogs developed lordoscoliosis with obvious rotation and the curves mimic AIS three dimensionally well. This method allows lumbar scoliosis to develop without deep dissection of muscle and maintains the essential anatomical elements along the spinal curve. Moreover, the spinal growth modulation technique could yield information that would provide a basis for developing novel early-stage treatments for children with scoliosis.