Objective To establish the mouse bone marrow-derived dendritic cells expressing FasL protein and explore the mechanism of inducing allogeneic mouse spleen T lymphocyte apoptosis. MethodsMouse myeloid DCs were cultured in selective medium zontaining essential cytokines for DC growth in vitro. The mouse DCs were transfected with liposome-mediated FasL gene. The levels of FasL mRNA before and after transfection were assayed by real-time quantitative PCR. The expression levels of FasL protein were assayed by flow cytometry(FCM)and Western blot. Non-transfected DC，empty plasmid transfected DC and FasL transfected DC were infused intravenouslY into allogeneic mouse. After 7 days, the apoptosis in spleen T lymphocytes was evaluated by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling)method and FCM. ResultsCultured in vitro, the mature myeloid DCs from mouse could be obtained.The expressions of FasL mRNA and protein in FasL transfected DCs were significantly higher. Through the detection of spleen T lymphocyte apoptosis with TUNEL，the apoptosis index(AI)was higher in FasL transfected DC(11.67±1.53)，compared with non-transfected DC(2.67±0.58)and empty plasmid transfected DC(3.33±0.58)，P＜0.01. ConclusionA large quantity of myeloid DCs can be obtained through in vitro culture in selective medium. The liposome-mediated FasL gene transfected DCs could successfully express high levels of FasL protein. Intravenous infusion of FasL gene transfected DCs could induce apoptosis of allogeneic mouse spleen T lymphocytes.

Objective KIF18A is a protein that has close relation with mitotic regulation and tumor development.This study aimed to establish KIF18A protein expression system in baculovirus,which may help to realize high efficient synthesis of KIF1 8A protein in vitro.Methods Transfer vector was constructed with molecular cloning method,and recombinant baculovirus were obtained through gene transposition.KIF18A protein was expressed by transforming recombinant baculovirus into infected insect Sf-9 cells to realize the synthesis efficiency.Results It was confirmed by DNA sequencing,microscopic observation and Western Blot that the KIF18A protein expression system in baculovirus was successfully established.Conclusions Conditions for transfer vector and recombinant virus transfection are defined,and high efficient KIF18A protein baculovirus expression system are successfully constructed.

Objective To study the effects of flavone compound α-NF on CYP1B1 gene in transplanted nude mice tumor with human paclitaxel resistance ovarian cancer cell. Methods Using 24 nude models of orthotopic transplantation of human ovarian cancer, they were divided randomly into four groups: control group, α-NF group, paclitaxel group and combined group. RT-PCR and Western blotting method were used to detect CYP1B1 mRNA and protein expression. Results There was no significant difference in CYP1B1 mRNA expression between α-NF group, combined group and control group(P >0.05). Compared with control group, CYP1B1 protein expression was distinctly lower in α-NF and combined group (P<0.01). Conclusion α-NF has no effect on CYPIB1 gene in mRNA level, whereas α-NF down-regnlate, CYP1B1 expression of transplanted A2780TS nude mice tumor in protein level, thus α-NF has the effect on reversing paclitaxel resistance of A2780TS cell.

Humans ; Oral Medicine ; trends ; Osteogenesis, Distraction ; Prostheses and Implants ; Tissue Engineering

Objective To investigate the relationship between the total burden of cerebral small vessel disease (CSVD) and the outcomes in patients with large artery atherosclerotic (LAA) stroke.Methods From June 2016 to January 2018,patients with LAA stroke treated at the Department of Neurology,the Affiliated Hospital of Qingdao University were enrolled retrospectively.The overall burden of CSVD was evaluated according to MRI findings.The National Institute of Health Stroke Scale (NIHSS) was used to evaluate theseverity of stroke.The modified Rankin scale (mRS) was used to evaluate the outcomes at day 90 after the onset.The mRS score 0-2 was defined as good outcome,and ＞2 was defined as poor outcome.Results A total of 148 patients with LAA stroke were enrolled,including good outcome in 72 (48.65％) and poor outcome in 76 (51.35％).There were significant differences in the proportions of hypertension (69.44％ vs.85.52％;x2 =5.519,P =0.019),taking antihypertensive drugs before the onset (48.61％ vs.69.74％;x2 =6.845,P =0.009),white matter hyperintensity (18.06％ vs.39.47％;x2 =8.228,P =0.004),enlarged perivascular space (33.33％ vs.60.53％;x2 =10.968,P =0.001),as well as the baseline NIHSS scores (3.00 [2.00-4.00] vs.7.0 [5.0-10.0];Z =-8.159,P =0.001),baseline systolic blood pressure (149.40± 15.80mmHgvs.157.21± 14.05mmHg;t=3.180,P=0.002;1 mmHg=0.133 kPa),fasting glucose (5.91 ±2.06 mmol/L vs.6.92 ±2.65 mmol/L;t =2.595,P =0.010),and the proportions of total CSVD scores 0,1,2,3,and 4 (Z =-4.927,P =0.001) between the 2 groups.After adjustment for the confounding factors,such as hypertension and fasting glucose,multivariate regression analysis showed that the total CSVD score (odds ratio 4.457,95％ confidence interval 1.768-11.236;P =0.002) and baseline NIHSS score (odds ratio 2.070,95％ confidence interval 1.580-2.710;P ＜ 0.001)were the independent risk factors for the poor outcomes in patients with LAA stroke.Conclusions The total CSVD burden was closely associated with the outcomes in patients with LAA stroke.Higher CSVD total score and baseline NIHSS scores were independently associated with the poor outcome at 90 d in patients with LAA stroke.

AIM: To study effect of endogenous carbon monoxide on intracellular calcium concentration and explore the mechanism in brain protection of endogenous CO in focal cerebral ischemia in rats. METHODS: SD rats were divided into three groups randomly, which including hemin, ZnPP group and saline group as control. Respectively saline, hemin, ZnPP were injected intra-peritoneally twelve hours before middle cerebral artery was occluded. Twenty four hours after MCAO model was set up, the concentration of carbon monoxide in blood and intracellular calcium in neural cells was examined. RESULTS: Contrast to saline group, the concentration of CO in blood rose up while intracellular calcium in occluded side decreased in hemin group; the concentration of CO in blood went down while intracellular calcium in occluded side rose up in ZnPP group, there was significant difference among them (P0.05). CONCLUSIONS: It may be one of mechanisms on brain protection in ischemic cerebral tissue that carbon monoxide affected intracellular calcium concentration of neural cells by regulating Ca~(2+)-K~+ channel on cell membrane as a messenger gaseous molecular and neurotransmitter. [

OBJECTIVETo discuss arthroscopic technique of single Kirschner wire and suture avoiding epiphyseal line fixation for tibial intercondylar eminence fracture and its clinical results.

METHODSFrom May 2008 to December 2012, 21 patients (13 males, 8 females, ranging in age from 6 to 14 years old) with tibial intercondylar eminence fracture were treated arthroscopically with single Kirschner wire and suture avoiding epiphyseal line fixation technique. According to Meyers and McKeever classification, 7 patients were type II, 10 patients were type III, and 4 patients were type IV. Active rehabilitation began at one week after operation. The patients were followed up for 10 to 30 months. X-ray films were taken to evaluate fracture healing at 1, 3, 6 months after operation; range of motion, the anterior drawer test, the lachman test and the Lysholm knee score were used to evaluate clinical effects.

RESULTSAll fractures were healed without displacement at 6 weeks after operation. Anterior drawer test and the lachman test were both negative in all patients at 3 months after operation. Lysholm knee score was 95.5 ±2.5 at 6 months after operation, and postoperative X-ray film did not find epiphyseal line broadening or narrowing.

CONCLUSIONArthroscopic treatment for tibial eminence intercondylar fracture with single Kirschner wire and 8-shaped suture avoiding epiphyseal line fixation technique has many advantages, such as firm fixation,early mobilization, less invasive, less injury of physis and satisfactory effect.

Adolescent ; Arthroscopy ; Bone Wires ; Child ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Sutures ; Tibial Fractures ; surgery

Equipment Design ; Facial Injuries ; therapy ; First Aid ; instrumentation

Objective To assess the clinical significance of modular femoral component in restoring femoral offset after total hip arthroplasty.Methods Data of 75 patients,who had undergone THA with modular femoral component from June 2009 to June 2012,were retrospectively analyzed.There were 42 males and 33 females,aged from 44 to 83 years (average,64.7±12 years).There were 40 cases of left hip and 35 cases of right hip.The average Harris hip score was 54±6 (range,32 to 65) before operation.The Zimmer M/L Taper hip prosthesis with Kinectiv technology was used in all patients.Preoperative templating was utilized to accurately predict the prosthetic size and the femoral offset.During the surgery,the femoral offset was measured accurately,and the appropriate femoral neck prosthesis was selected in order to restore the femoral offset and reconstruct hip abductor moment arm.The femoral offset and leg length were measured according to X-rays after operation.The Harris hip score was used to assess the clinical results.Results The femoral offset was reconstructed in all patients,ranging from-3.0 to 3.0 mm (average,1.3±0.3 mm).The average leg length discrepancy was improved to 3.0±0.8 mm.All patients were followed up for 6 to 36 months (average,19.7 months).The average hip abduction angle was 40°±10° at 12 weeks after operation.The average Harris hip score was 86±13 (range,65-94) at 12 weeks after operation and 95±15 (range,78 to 98) at final follow-up.There was no hip dislocation and femoral acetabular impingement syndrome.Conclusion Modular femoral component can restore femoral offset without changing the leg length,which can result in good hip abductor strength,range of motion of the hip and stability,as well as less complications.

BACKGROUND:Due to the importance of pedicle adjacent structures, once the screw replacement appears a deviation, adjacent structures may be damaged, leading to extremely serious consequences. Although the security of screw placement in thoracic vertebral pedicle-rib complex is significantly greater than that of pedicle screws, the mechanics of the pedicle-rib complex at different cross-sectional areas of the screw are rarely reported.
OBJECTIVE:To observe mechanical property of different cross-sectional area screws in the middle and upper thoracic vertebral pedicle-rib complex.
METHODS:Five specimens of adult cadaveric thoracic spine (T 1-T 10 ) and adjacent rib segment (50-60 mm long) were used. The bone density of specimens was measured using difunctional bone density testing machine, and osteoporotic vertebral body was excluded. The position of the screws was detected with CT images. The maximal withdrawal force of the pedicle screw was measured with biomechanical force test machine.
RESULTS AND CONCLUSION:Thirty-eight specimens at normal bone density were implanted with 25 screws (5.5 mm), 25 screws (6.0 mm) and 26 screws (6.5 mm). Because the pedicle screws destroyed the pedicle-rib complex and perforated the vertebral body, we final y obtained the withdrawal force of 68 screws. The axial withdrawal force of pedicle screws at different diameters was (812.36±147.22) N, (868.64±160.48) N and (946.48±157.58) N, respectively. There were significant differences between the 5.5 mm screws and the 6.5 mm screws (P<0.05). Experimental findings indicate that, the pedicle screws (diameter>5.5 mm) are suitable in the middle and upper thoracic vertebral pedicle-rib complex due to strong internal fixation and clinical requirement.