Analysis Of The Mechanism How Intervertebral Disc Degeneration Delayed By Estrogen And Analysis Of The Classification And The Imaging In Patients With Scoliosis

Chapter 2(Section 1)Does Estrogen Protect Lumbar Intervertebral Disc from Degeneration through Autophagy in Ovariectomized Rats?Background.Intervertebral disc degeneration is the initiating factor of degenerative diseases of the spine,which can lead to degenerative scoliosis,disc herniation and other diseases,and has a great impact on the quality of life of the patients.The disc degeneration progresses more rapidly in postmenopausal women.Previous studies have shown that estrogen can regulate autophagy in other tissues such as the brain,heart,and liver.This study aimed to investigate whether estrogen slow intervertebral disc degeneration by regulating the level of autophagy in the intervertebral discMethods.Sixty 3-month-old female Sprague-Dawley rats were randomly divided into sham group(sham operation group),control group(OVX group),estrogen group(OVX+E group)and autophagy inhibition group(OVX+E+3-MA group)with 15 rats in each group.All the SD rats received ovariectomy except for group A.The rats were injected with different drugs.Group A and group B were injected subcutaneously with olive oil.Groups C and D were injected subcutaneously with 17β-estradiol.Groups A,B and C were injected intraperitoneally with normal saline,and group D was injected intraperitoneally with 3-MA.The rats were injected 5 times a week continuously for 16 weeks.Spinal CT scans were performed after anesthesia in rats.After the rats were sacrificed,L1/2,L2/3,and L3/4 disc nucleus pulposus tissues were obtained surgically.Intervertebral disc degeneration-related proteins and autophagy-related proteins were detected by Western Blot.L5/6 disc nucleus pulposus tissues were obtained to do electron microscope examination.The transmission electron microscope scans the autophagic bodies in the nucleus pulposus.The L4 and L5 vertebral bodies and L4/5 intervertebral discs were preserved together.After decalcification,they were embedded in paraffin,and sectioned to detect degeneration-related proteins and autophagy-related proteins by immunohistochemistryResults.There was no difference in disc height,vertebral height,and endplate integrity between the four groups.Western Blot results showed that the expression of Col2α in nucleus pulposus cells of OVX group was lower than that of Sham group.The expression of Col2α in OVX+E group was higher than that in OVX group.The expression of Col2α in OVX+E+3-MA group was lower than that in Sham group and OVX+E group.The expression of Beclin-1 in OVX group was lower than that in Sham group.The expression of Beclin-1 in OVX+E group was higher than that in OVX group.The expression of OVX+E+3-MA group was lower than that in Sham group and OVX+E group.H.E.staining showed that there were more nucleus pulposus cells in Sham group,and the number of nucleus pulposus cells in OVX group was less than that in Sham group.The number of nucleus pulposus cells in OVX+E group was more than that in OVX group,and the number of nucleus pulposus cells in OVX+E+3-MA group was less than Sham group and OVX group.The expression of Col la in Sham group was lower,and the expression of Col la in OVX group was higher than that in Sham group.The expression of Col la in OVX+E+3-MA group was higher than that in OVX+E group and Sham group.The expression of Beclin-1 and LC3 in Sham group was higher than that in OVX group.The expression in OVX+E group was higher than that in OVX group.The expression of Beclin-1 and LC3 in OVX+E+3-MA group was lower than that in Sham group and OVX+E group.Electron microscopy showed that there were more autophagosomes in the Sham group and the OVX+E group,while there were fewer autophagosomes in the OVX group and the OVX+E+3-MA groupCONCLUSION:After ovariectomy in rats,the number of nucleus pulposus cells in the disc decreases,the expression of Col la increases,and the expression of Col2αdecreases.This indicates that the intervertebral discs of rats have degenerated,and estrogen replacement can improve the degeneration of intervertebral discs.The expression of Beclin-1 and LC3 in the nucleus pulposus cells of the ovariectomized rats was reduced,and the autophagosomes were reduced.After estrogen supplementation,Beclin-1,LC3 and autophagosomes were increased again,indicating that estrogen can promote autophagy of the nucleus pulposus in rat intervertebral discs.In the ovariectomized rats supplemented with estrogen and injected with autophagy inhibitors,intervertebral disc degeneration aggravated,indicating that in rats,estrogen can slow the intervertebral disc degeneration by promoting autophagy.Chapter 3(Section 1)Could Pelvic Parameters Determine Optimal Postoperative Thoracic Kyphosis in Lenke Type 1 AIS patients?Background.A proper restoration of sagittal alignment is essential in AIS(adolescent idiopathic scoliosis)patients,but few studies provided a formula to predict an optimal surgical thoracic kyphosis(TK)gain in AIS patients.A formula was recently proposed(LL=(PI+TK)/2+10)to predict the optimal lumbar lordosis(LL)in adult spinal deformity patients,which has not been validated in adolescents.The aim of this study is to establish a formula with TK and pelvic parameters in normal adolescents and predict an optimal TK with this formula pre-and post-operatively in Lenke 1 AIS patientsMethods.A total of 60 asymptomatic adolescents were used to validate the proposed formula.The subject was considered to match with the formula,if the difference between the virtual TK and the theoretical TK was less than 10°.Then regression analysis was performed to establish a new formula to predict TK in adolescents.The predictive efficiency of the new formula was also validated in 40 Lenke 1 AIS patients.Results.Of the 60 asymptomatic adolescents,only 26(43.33%)asymptomatic adolescents matched with the adjusted formula:TK=2×(LL-10)-PI.The paired t test revealed a significantly different theoretical TK(tTK)compared to the virtual TK(41.23±18.29°vs.24.80±8.75°,P<0.001).Multiple linear regression showed that TK had a relationship with LL,SS and age(R2=0.331):TK=-0.785×LL-0.843×SS+0.858×age+3.754.There were 27(67.50%),32(80.00%)and 35(87.50%)Lenke 1 AIS patients matched this formula preoperatively,postoperatively and at the last follow-upConclusion.Our results revealed that the predictive formula for sagittal alignment for adults was not applicable in adolescents.This study established a new predictive formula for TK based on asymptomatic adolescents.In Lenke 1 AIS patients,post-op TK in 87.5%of patients matched the predictive value,indicating that the new formula can be considered as a reference when making a surgery strategyChapter 3(Section 2)Will Etiology Affect the HRQoL in ASD Patients with Different SRS-Schwab Classification Sagittal Modifiers?Background.Differentiating etiology is important for surgical decision,while SRS-Schwab classification did not encounter it.This study aims to investigate whether etiology is related with HRQoL(health related quality of life)in patients with different sagittal modifiers of SRS-Schwab Classification.Methods.79 ASD patients were divided into 2 groups based on etiologies:idiopathic scoliosis(18 females,11 males;mean age 33.7 years)and de novo scoliosis(29 females,21 males;mean age 62.1years).HRQoL data including SRS-22,SF-36 and ODI scores were obtained.ODI scores were adjusted using the offset of raw value from normative value of different age to eliminate the impact of age.ANOVA of each modifier grade was performed within etiology group and t test between etiology groups.Spearmen correlation was also performed.Results.IS group saw significantly better SF-36(59.9 vs.41.6,p<0.01)and ODI scores(20.67 vs.40.5,p<0.01)compared to DS group,and also in SF-36 PCS and MCS.ODI offset from normative value also supported worse HRQOL in DS patients(8.8 vs.18.0,p=0.03).Within IS group,there was a significant difference of SF-36 score found across different modifier levels(0,+,++)in PI-LL modifiers(62.31 vs.73.96 vs.24.04,p=0.03),which was not seen in DS group.IS patients often showed better HRQoL scores than DS patients with similar modifier grade(Table 1).ODI-offset was better in IS patients with grade 0 PI-LL(4.77 vs.18.01,p=0.065),grade 0 SVA(8.47 vs.18.32,p=0.05)and significantly better in IS patients with grade 0 PT(8.75 vs.24.64,p=0.01).DS patients had worse SF-36 scores in all 3 modifiers.Conclusion.Etiology may be unneglectable when considering SRS-Schwab classification.HRQoL of IS paitents is much better than DS patients with similar SRS-Schwab classification sagittal modifiers.This result also indicates the involvement of etiology in surgical decisionChapter 3(Section 3)Thorax asymmetry in AIS patients:Will it improve after brace treatment?Background.Adolescent idiopathic scoliosis(AIS)is a common spinal deformity Bracing is the only community-recognized conservative treatment for AIS patients.The aim of this study is to determinate if the asymmetry of thorax and breast would aggravate after brace treatment.Methods.Twenty Lenke type 1 AIS girls with brace treatment history and 20 matched patients without brace treatment were analyzed.The Cobb angle and the apical vertebra were matched between the two groups by design.Five morphological parameters were measured on topless photographs and 11 radiographic parameters were obtained on the preoperative CT scans.The RAml angle and the difference between the parameters of the concave side and the convex side was utilized to describe the extent of the thorax and the breast asymmetryResults.The average age was not significantly different between the brace and non-brace group(15.3 years.and 16.1 years.,P=0.169).The Cobb angle and the apical vertebra was well matched(P=1.000 and P=0.478).The difference of the sternal notch-nipple length(SNL)between the concave side and the convex side in the brace group was significantly smaller than that in the non-brace group(-2.5±8.7mm vs.-12±9.1mm,P=0.001).Other morphological and radiographic parameters showed no differenceConclusion.Our results reveal that the brace treatment won’t aggravate the thorax and breast asymmetry in AIS patients,and might has some improving effects on the breast instead.