Objective To gain purified platelets for further research of the immune function of platelet.Methods Platelets were purified by positive(MS) and negative(LD) magnetic cell sorting(MACS) separation.The percentage of activated platelets was detected by flow cytometry and nucleated cell clearance was evaluated by white cell count.Results The percentage of activated platelets before separation was(2.39?1.10)%,and increased to(2.56?1.08)% and(16.76?4.04)%,respectively,after MS and LD MACS.The clearances of nucleated cells after MACS MS and LD were(98.44?0.24)% and(98.47?0.18)%,respectively.The recovery rate of purified platelet after MS and LD was(76.50?1.49)% and(79.20?2.61)%.Conclusion MACS LD method was more suitable for the platelet purification.

Objective To investigate cause,diagnosis,treatment and prevention of vertebral artery injury in cervical spine surgery.Methods Data of 7 patients with vertebral artery injury caused by cervical spine surgery from October 2002 to April 2012 were retrospectively analyzed.There were 6 males and 1 female,aged from 23 to 65 years (average,48.9 years).The reasons of cervical spine surgery were as follows:cervical spondylotic myelopathy (5 cases),traumatic subluxation of C4 and C5 (1 case),and cervical spinal stenosis due to skeletal fluorosis (1 case).All cases had unilateral vertebral artery injury,including 4 cases in the left side and 3 cases in the right side.The cause,treatment and prognosis of vertebral artery injury in 7 patients were analyzed.Results Four patients suffered from vertebral artery injury during anterior cervical spine surgery; 2 cases were caused by excessive lateral trepanation,1 by excessive lateral discectomy using a curette,and 1 by Kerrison punch during decompression in a patient with anomalous vertebral artery due to traumatic subluxation of C4 and C5.Three patients suffered from vertebral artery injury during posterior surgery; 2 cases were caused by a drill bit during drilling lateral mass of C4 and 1 by a rongeur during removing posterior C1 arch in a patient with cervical spinal stenosis due to skeletal fluorosis.After vertebral artery injury,all patients were immediately treated by hemostatic tamponade,and then the incisions were closed.However,1 patient occurred delayed cervical hemorrhage 4 weeks after operation.Then he underwent vertebral artery angiography,balloon embolization and evacuation of hematoma.All cases had no cerebral infarction.However,two of them had postoperative temporary dizziness.Conclusion Vertebral artery injury in cervical spine surgery is a rare but serious complication,which is associated with surgical mistakes and anatomical variations.Hemostatic tamponade and embolization are effective for this complication.

Objective:To study on significance of ABO genotyping.Methods:To use the methods of polymerase chain reaction sequence specific primers(PCR SSP) for genotyping of ABO blood group and observation ABO gene polymorphism as well as identification of doubt sample.Results:The reliability of the genotyping for ABO blood group was proved by testing DNA samples previously known genotypes.The results of genotyping for 104 healthy and unrelated HAN individuals were correspond with serological phenotypes.The method of ABO genotyping was applied for clinical pre transfusion ABO typing,pre delivery fetal ABO typing,parenting test and subgrouptyping.Conclusion:The method of ABO genotyping may correct typing for doubt sample of ABO serological typing.

Objective To analyze the change of phosphotidyl serine(PS) in apheresis platelets(plts) during storage process,and to evaluate the quality of the plts during the storage period.Methods The apoptosis ligand Fas(CD95) was used to induce plts as apoptosis model,and by which the method to detect plts PS by flow cytometry(FCM) was established.Subsequently the resting PS expressive rate and the induced PS expressive rate of 30 apheresis plts,which induced by CD95,were determined using the FCM method established in storage process.Results At the 1st,5th,7th,8th day of storage process,the PS expression of plts kept on increasing.The PS expressive rate of plts were 2.03%?0.91%,3.26%?1.86%,8.98%?4.60%,26.68%?21.28%,and 38.85%?26.43%,respectively,and there were significant differences among groups respectively(P0.05).Conclusion The PS expression of apheresis plts increase significantly with the prolongation of storage time;Fresh plts has the ability of anti-apoptosis;Apheresis plts could keep normal function until storage 7 d.

Objectives To investigate influencing factors and pattern of posterior shifting of the spinal cord after cervical laminoplasty.Methods Forty three patients with compressive cervical myelopathy,including 32 males and 11 females,aged from 33 to 78 years (average,57.9 years),treated with open-door laminoplasty from C3 to C7 between February 2008 and October 2010,were enrolled in this study.There were 30 cases of cervical spondylotic myelopathy with developmental cervical stenosis and 13 cases of ossification of the posterior longitudinal ligament.Twenty one cases underwent modified open-door laminoplasty using anchor method,and the other 22 cases underwent laminoplasty with preservation of the unilateral muscular ligament complex.According to midsagittal T2-weighted MRI in the neutral position,several parameters indicating shift distance of the spinal cord and dural sac were measured at each level before operation and at 3 months after operation:shift of anterior margin of the spinal cord (SAMSC),shift of posterior margin of the spinal cord (SPMSC),shift of anterior margin of the dural sac (SAMDS) and shift of posterior margin of the dural sac (SPMDS).The line connecting the top of dens and posterior inferior angle of T1 was defined as E line and the length was E.The length of perpendicular line from each base point to E line was Px (x =1-6).The parameter (100×Px/E) was used to represent local curvature at each level,and the curvature index (CI)was used to represent the global curvature of the cervical spine.Then correlation analyses of the parameters above were performed.Results There was no significant change in SAMDS at different level after surgery,however SAMSC,SPMSC and SPMDS changed significantly and synchronously.At the level of C5 and C6,SPMSC was maximum,but not correlated to CI.Actually,SPMSC was correlated to the local curvature and highly correlated to SPMDS at the same level.Conclusion After cervical laminoplasty,posterior shifting of the spinal cord was highly correlated to posterior shifting of the dural sac at the same level,weakly correlated to local curvature and not correlated to the global curvature of the cervical spine.

Objective To quantitatively compare the effect of preservation or removal of atlas posterior arch on cervical posterior decompression,so as to provide a basis for reasonable selection of upper cervical spine decompression range and determination of surgical indications for atlas posterior arch resection.Methods The data of 45 patients with posterior decompression of upper cervical spine were retrospectively analyzed.According to the decompression range,the patients were divided into C2-C7 group and C1-C7 group.There were 25 cases in the C2-C7 group,19 males and 6 females,with an average age of 56.3 years (40-71 years),4 cases of cervical spondylotic myelopathy and 21 cases of ossification of the posterior longitudinal ligament of cervical spine.All of the 25 patients underwent open-door laminoplasty:20 cases with hinge side anchoring procedure and 5 cases with preservation of the unilateral posterior muscular-ligament complex procedure (titanium cable procedure).There were 20 cases in C1-C7 group,12 males and 8 females,with an average age of 58A years (44-75 years).All of the 20 cases underwent atlas posterior arch resection as well as C2-C7 open-door laminoplasty,including anchoring procedure in 1 case,titanium miniplate procedure in 4 cases,and titanium cable procedure in 15 cases.Standardized vertebral-cord distance (SVCD) at each level from atlas to level C~ was measured on T2-weighted images of MR on the mid-sagittal plane in the neutral position pedormed 3-12 months postoperatively at each individual level.As the main outcomes,the SVCD values obtained at the same level of the two groups were compared between the two groups.Shapiro-Wilk normality test was performed on the SVCD values at C1.2 and C2 levels of two groups.The area under the normal distribution curve of SVCDs was used to calculate the corresponding residual compression rate with different magnitude of compression mass to further discover the difference of the decompression effect between the two groups.Results The SVCD obtained at the level of the anterior arch of atlas (C1),the junction of odontoid process and axis (C1,2) and the middle part of axis body (C2) in the C2-C7 group was 9.91±1.34 mm,8.35±1.27 mm,and 8.22 ±1.43 mm,respectively.The SVCD at the same levels was 11.02±1.60 mm,9.72±1.24 mm,and 9.12±1.11 mm,respectively.SVCDs differed significantly in the above range between the two groups.However,from level C2,3 to C6,7,there was no significant difference in SVCDs between the two groups.The JOA score of group C2-C7 was 11.8±2.7 preoperatively and increased significantly to 14.7±1.8 at 12 months postoperatively(t=-7.006,P＜0.001) with a recovery rate of 57.0％±32.2％.The JOA score of group C1-C7 was 11.7±2.8 preoperatively and increased significantly to 14.2±2.3 at 12 months postoperatively(t=-6.177,P＜0.001) with a recovery rate of 51.9％±32.1％.Conclusion Atlas posterior arch resection can significantly increase the decompression effect of posterior cervical surgery from the anterior arch of atlas to the middle part of axis body,but it would not increase the decompression effect at level C2.3 or below.When the magnitude of the ventral compression factor exceeds the decompression limit (8.5 mm) available with C2-C7 decompression in the range from atlas to the middle of the axis body,extending the decompression range by atlas posterior arch resection is an effective means to achieve adequate decompression.

Objective　To investigate the clinical efficacy of anterior cervical surgery by comparative analyzing hand dysfunction using brief Michigan hand questionnaire(Brief MHQ)in Hirayama disease patients.Methods　From Aug 2011 to Dec 2016,27 patients of hirayama disease who underwent surgery were enrolled in this study.The study group consisted of 27 men.The mean follow-up period was 41.1 months.The levels of surgery included 18 cases of C4-C7,6 cases of C3-C6,2 cases of C4-C6 and 1 case of C5-T1.Brief MHQ were evaluated for the 27 patients.According to the Wilcoxon analysis,the unchanged domains were analyzed with the multifactor Logistic regression analysis by preoperative duration of symptoms,age of onset,and number of affected extremities.Dynamic flexion-extension lateral X-rays were performed at baseline and at final follow-up.Results　No failure of internal fixation was detected on dynamic flexion-extension lateral X-rays.Five domains of preoperative Brief MHQ had lower scores,including Function,Satisfaction,Aesthetics,Activities of daily living,Work domain.With the exception of Aesthetics and Pain domain,all the other four domains showed significant improvement after surgery.The total score was 38.44±5.83 at base-line and 43.19±4.47 at follow-up.The score of Function was 5.19±1.36 at baseline 6.37±1.15 at follow-up;The score of Satisfaction was 5.56±1.22 at baseline 6.60±1.05 at follow-up;The score of Activities of daily living was 6.33±1.84 at baseline 7.60±1.47 at follow-up;The score of Work was 6.85±1.75 at baseline 7.67±1.33 at follow-up.The risk factors of postoperative outcomes reported in the literature included duration of disease at the time of surgery,age of onset,and extremity involvement.According to the Logistic regression,pre-operation duration was the risk factor for Aesthetics domain and the cut-off time was 1.75 years.Conclusion　Four domains of Brief MHQ score were improved significantly after anterior surgery for patients with hirayama disease.Brief MHQ was useful to evaluate the hand dysfunction and clinical efficacy in patients with hirayama disease.

【Objective】 To investigate the effect of ssDNA aptamer of RhD blood group antigen on erythrocyte toxicity. 【Methods】 Two full-length ssDNA aptamers(82 bp) of RhD blood group antigen were obtained by gene synthesis.Five samples of whole blood with EDTA anticoagulant were collected to prepare red blood cell suspensions (4×10⁷/mL), which were split into 10 tubes(100 μL/tube), corresponding to 5 experimental groups and 5 controls.Two monospecific full-length ssDNA sequences (100 pmol/μL, 5μL each) were added into the experimental group, while the same amount of normal saline into the control.After treatment, the experimental group and the control were incubated for 60 min at 37℃.After washing, they were suspended in LISS solution and stored at 4℃.The experimental group and the control were set according to different time point during storage (1 h, 1 d, 3 d, 10 d and 17 d), with 5 tubes in each group.For erythrocytes in LISS suspension at different storage time, Annexin V labeled with FITC was used as a probe to label the phosphatidylserine (PS) content and Fluo-4 to label Ca²⁺ .The eversion of PS and the change of Ca²⁺ concentration in red blood cells in LISS suspensions were determined by flow cytometry. 【Results】 After incubation, all groups were examined under the light microscope.No agglutination occurred in the experimental group, while agglutination occurred in the control.Flow cytometry showed that the number of Annexin V-FITC staining cells of suspended erythrocytes at the same storage time-point was similar between the experimental group and the control, with no significant differences.In the experimental group, apoptosis rate of Annexin V cells at 10-day storage(6.06±1.38) was significantly higher than that at 1-hour storage(P<0.05), so as at 17-day storage(7.77±1.23) than 1-hour, 1-day and 3-day storage(P<0.05). The apoptosis rate of Fluo-4 AM cell in suspended RBCs at the same storage time-point was similar between the two groups(P>0.05). In the experimental group, the apoptosis rate of Fluo-4 AM cell at the 3-day, 10-day and 17-day storage was 20.84±4.16, 22.35±3.37 and 27.06±2.81, respectively(P<0.05). 【Conclusion】 ssDNA aptamer was not found to have any cytotoxic effects on red blood cells, and RhD ssDNA aptamer may be used as a material for the detection and preparation of universal blood.

Drug-induced immune hemolytic anemia (DIIHA) is a rare cytopenia caused by damage to RBCs by drug-induced antibodies or non-immune protein adsorption (NIPA). The drugs associated with DIIHA and the mechanistic hypotheses that are thought to be involved have been controversial, with complex serological tests often required by specialized Immune Hematology laboratories for diagnosis. It is necessary to know the clinical manifestation and laboratory diagnosis of DIIHA in order to distinguish the immuno-hematological abnormality caused by drugs from other causes. How to improve the diagnostic ability of DIIHA and establish a scientific and reasonable idea of DIIHA serological examination is urgent to help clinical diagnosis and correct treatment.

Objective:To explore the safety, feasibility, and short-term outcome ofdistraction on the concave side in the treatment of patients with congenital cervical scoliosis.Methods:Between August 2015 and December 2019, 11 patients with congenital cervical scoliosis underwent distraction technique on concave side, among which 5 were males and 6 were females. Age was 9.9±3.1 years old (range 6-16 years old). The primary cervical spine deformity was hemi-vertebra with different degrees of vertebra fusion. 7 cases were in C 3, 3 cases were in C 4 and 1 case was in C 5. Anterior-posterior combined approach was used. Firstly, discectomy and soft tissue release on concave side were made through anterior approach, then distraction on concave side and fusion with internal fixation were made through posterior approach and at last fixation and fusion in anterior approach were made. In this study we measured structure Cobb angle, compensatory Cobb angle, mandibular incline, shoulder balance and the angle difference of trapezius muscle preoperation and post operation. Perioperative neurological,vascular and wound related complicationswere recorded. Results:All patients' surgeries were completed successfully. Eight patients received single site distraction and 3 patients received distraction in two sites. The duration of surgery was 466±141 min (range 150-659 min), the estimated blood losswas 387±191 ml (range 100-660 ml) and follow up time was 12.2±9.5 months (range 3-24 months). Structural Cobb angle was 28.9°±13.1° pre-operation and 7.4°(3.0°, 27.7°) post-operation at 3 months, which was corrected significantly ( Z=-2.934, P=0.003). The correction rate was 58.1±26.1% (range 18.8%-97.6%). Structural Cobb angle was 13.2°±12.3°at 1 year post operation and had no significant difference compared with 3 months post operation ( t=1.960, P=0.107). Compensatory Cobb angle was 18.3°±6.1° pre-operation and 9.4°±7.3° post-operation at 3 months, which was corrected significantly ( t= 5.071, P<0.001) and the correction rate was 51.3%±28.3% (range 2.4%-94.7%). Compensatory Cobb angle was 8.9°±7.7° at 1 year follow up and was corrected significantly ( t=5.253, P=0.003) compared to 3 months after surgery and the correction rate was 61.4%±26.9%. Two patients developed C 5 nerve root dysfunction and 1 patient developed numbness on the index and middle fingers after surgery. All of them occurred on the concave side and recovered by conservative treatment. Conclusion:The application of distraction on the concave side in the treatment of congenitalcervical scoliosis is with good feasibility and clinical safety. Short-term follow-up showed excellent resultswith a promising future.