Expression of Caspase-4 and Caspase-3 Gene Related Apoptosis in Pathological Changes of Sternocleidomastoid of Congenital Muscular Torticollis and Four Years Clinical Followed-up for Opereated Patients
|School||Zunyi Medical College,|
|Keywords||congenital muscular torticollis sternocleidomastoid (SCM) muscles Caspase-4 Caspase-3 Apoptosis immunohistochemistry follow up sternocleidomastoid muscle|
Part IExpression of Caspase-4 and Caspase-3 Gene Related Apoptosis in Sternocleidomastoid of Congenital Muscular Torticollis Shenzhen Children’s Hospital, Zunyi Medical College Wang Shuaiyin (Candidate of MD) Tang Shengping (Supervisor)Objeetive To detected protein expression of Caspase-4 and cleaved Caspase-3 in sternocleidomastoid of congenital muscular torticollis and to explore the possible mechanism of apoptosis in the pathogenesis of CMT.Material and Methods From 2005 to 2010, a total of 73 cases of CMT were operated in Department Pediatric Orthopedics, Shenzhen Children’s Hospital. All samples were harvested during operation and were processed for HE stain, Masson and immunohistochemical stain for cleaved Caspase-3 and Caspase-4. Pathological changes of sternomastoid muscle section stainmed by HE staining were observe. The ratio of muscle to collagen were determined by Masson collagen stainning method. Each slice were reviewed under the 10×10 light microscope by using color image analysis system (ImagePro-6.0 image analysis system) for caculating area of muscle fiber and collagen fiber. Patients were grouped by age and different ratia of collagen to mucle.. Immunohistochemical method was performed to investigate the expression of Caspase-4 and cleaved Caspase-3Results The age distribution were as follow:3 months-<6 months,18 cases;7-≤12months,30 cases; 1 year-≤3 years,16 cases,≥4year-≤11years,9 cases. By HE staining,the basic abnormality is endomysial fibrosis with deposition of collagen and migration of fibroblasts around individual muscle fibers that undergo atrophy.Light have shown replacement of the sternocleidomastoid muscle by dense fibrous tissue without evidence of hemorrage and infection.Histologic have demonstrated degeneration of muscle fibers.Statistic proce-dure:all cases were grouped by age and tumor conditions.By Masson collagen staining method, it was observed that the collagen fibers proliferated excessively.All specimens were found in a large sheet of fibrous tissue proliferation in varying degrees and some bundles of fibrous tissue can be seen in them. CSA of skeletal musele fibers in the difference age group as follow:3 months-<6 months, 0.79±0.10;7 months-≤12months,0.65±0.17; 1 year-≤3 years,0.69±0.16,3 years-≤11years, 0.67±0.15, has significant difference (P<0.05).3 months-<6 months is greater than 7 months-≤12 months and 1 year-≤3 years and 3 years-≤11 years, has significant difference(P<0.005). Result of immunohistochemistry:Expression of cleaved Caspase-3 in he difference age group as follow:3 months-≤6 months,11.46±1.75;7 months-≤12months,10.33±6.01; 1 year-≤3 years,8.75±2.84,>4years-≤11 years,5.13±3.07 (p<0.05).3 months-≤6 months and 7 months-≤12 months is greater than 3 years-≤11 years, has significant difference(P<0.005). Expression of cleaved Caspase-3 in different CSA of skeletal musele fibers groups as follow: 30%-<50%type group 6.46±3.18.50%-≤70% type group 6.46±3.18,>70% type group 11.10±4.11 (p<0.05).>70% type group is greater than 30%-<50%type group and 50%-≤70% type group, has significant difference(P<0.01).Expression of Caspase-4 in he difference age group as follow:3 months-≤6 months,8.97±5.02;7 months-≤12months, 14.34±5.29; 1 year-≤3 years,9.02±4.37,≥4years-≤11 years,7.94±2.71, has significant difference (P<0.05).7 months-≤12 months is greater than 3 months-≤6 months and 3 years-≤11 years and 1 year-≤3 years, has significant difference (P<0.005).Expression of Caspase-4 in different CSA of skeletal musele fibers groups as follow:30%-<50%type group 9.93±3.49,50%-≤70% type group 10.76±3.42,>70% type group11.84±4.65,has significant difference (P<0.05).>70% type group is greater than 30%-≤50% type group, has significant difference(P<0.01).Conclusions 1、The level of stromal hyperplasia, or fiber in sternocleidomastoid muscle of CMT may be reduced gradually with age in the children less than one year old, but it may gradually increased with increased age after one year old.2、The apoptosis do exists in the interstitium in SCM and could happen to interstitial cell, muscle cells and musclular satallisal cells. More apoptosis cells were found in interstitium.The level of apoptosis may be reduced gradually with age in interstitium and increased with the increasing of fibrosis. The muscles fibrosis are found in more than one year children with CMT may infer the reason why some CMT can not be treated conservatively. The muscle fibrosis are possiblely related to apoptosis of the muscle satellite cells. The more muscle fibrosis, the severe the apoptosis of satellite cells. 3、The apoptosis pathway of cells in CMT may be different. Caspase-dependent pathway may be related to apoptosis of the interstitium cells. Endoplasmic reticulum pathway may be related to apoptosis of muscle cells or muscle satellite cells.Part II Functional and cosmetic analysis after surgery in congenital muscular torticollisObjeetive The aim of this study was to assess status after surgery for congenital muscular torticollis.Material and methods We investigated a total of 73 patients who came from department of pediatric orthopedics,Shenzhen Children’s Hospital.ZunyiMedical College. Ineluding 73 cases of congenital muscular torticollis diagnosed clinieally and pathologieally.A detailed questionnaire was applied to 65 patients who were operated for congenital muscular torticollis by phone call and interview.Sixty-five children were evaluated once and the scoring system used included functional and cosmetic results. Neck movement, muscle strength and endurance were compared with the uninvolved side. Head tilt, elevation of the shoulder, craniofacial asymmetry, and lateral band and operative scar were evaluated by clinical observations and photographs.65 cases of them were followed up on the purpose of prognostic analysis.Finally the clinical,pathological features and prognosis of all the investigated patients were analyzed retrospectively.Results In the 73 cases of congenital muscular torticollis,there were 43 male and 22 female. The age of onset ranged from 3 to 11 years,with the median age of 11 months.13 patients underwent surgery before they were 6 months of age.29 between 7 and 12 months of age,17 between 1 and 3 years of age,6 between 4 and 11 years of age.They were followed up on between 1 and 5 years,with an average of 4 years.51 had excellent results(78.4%) and 6 had good results(9.2%),8 fair(12.3%),and 0 poor,0% required a second operation.①For the age>3 Months-≤12 Months children,38 had excellent results and 2 had good to excellent results,3 fair,and 0 poor,0% required a second operation.②For the age>l years～≤3 years children,12 had excellent results and 3 had good to excellent results,2 fair.and 0 poor,0% required a second operation.③The age>3 years～≤11 years children 2 had excellent results and 1 had good to excellent results,3 fair.and 0 poor,0% required a second operation.indicating the benefit of surgery even in the late cases.Conclusions Method of distal unipolar open release and partial excision of the clavicular and sternal heads of the sternocleidomastoid muscle and and bipolar radical excision and partial excision was effective to treat congenital muscular torticollis. Most children achieved an overall excellent or good result after surgery. indicating the benefit of surgery even in the late cases.The small age children who was poor efficacy results by conservative treatment,early surgical treatment can help to improve the prognosis of CMT.