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

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. [

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.

Animals ; Fatigue ; prevention & control ; Female ; Gardenia ; chemistry ; Hypoxia ; prevention & control ; Male ; Mice ; Mice, Inbred Strains ; Plant Extracts ; pharmacology ; Thuja ; chemistry ; Ziziphus ; chemistry

Objective To investigate the association of body mass index (BMI),sex and age with blood pressure in non-physical labor population from Beijing area.Methods 14 237 subjects (67.06% male) ranged from 18.0-93.0 years (mean (SD):49.8(10.2)) wero recruited in the health examination in Beijing hospital. Statistical analysis wag performed using SPSS 12.0 software.Results The average BMI wag 22.30 ks/m~2 (SD: 3.08).Systolic blood pressure and diastolic blood pressure increaged with the increasing of BMI.The prevalence of hypertension in males <60.0 years wag considerably higher than that of female (24.28% and 13.77% for male and females, respectively, P=0.000).However,there were no significant statistical differences for the pmvalence between male and female older than 60.0 years (P=0.268).Multiple logistic regression showed that in males,using the group with BMI<18.0 kg/m~2 as reference group,in the groups of BMI 18.0-23.9,24.0-27.9,≥28.0 kg/m~2, OR wag 1.709 (95% CI:0.920-3.173),3.154(1.703-5.839),5.125 (95%CI:2.805-9.696),respeetively.In females.OR for hypertension in the groups of BMI 18.0-23.9,24.0-27.9,≥28.0 kg/m~2,OR was 1.988(95%CI:1.033-3.824),5.703(2.962-10.982),14.358(95%CI:7.334-28.106),respectively.Conclusions BMI is associated with hypertension.Prevention and eontrol of the risk factom of hypertension e.g.overweight and obesity is an important public health issue in China.

Objective To track the effects of hyperbaric oxygen (HBO) therapy on neonatal rats with hypoxic-ischernic brain damage (HIBD).MethodsA total of 126 healthy,seven-day-old SD rats were used to model the HIBD and then randomly divided into seven groups of 18:a sham group,a hypoxic-ischemic (HI) group,a 6 h HBO group,a 24 h HBO group,a 48 h HBO group,a 72 h HBO group and a 1 week HBO group.One hour of HBO treatment at 2 atmospheres absolute pressure was administered once daily for a week to the rats in the latter 5groups,starting at 6,24,48,72 hours and 1 week post the HIBD modeling,while no HBO was administered to the sham and HI groups.The effects were assessed in terms of histological changes ( the neuron density and the percentage of neuron apoptosis in the CA1 region of the hippocampus) and nitric oxide (NO),malondialdehyde (MDA) and superoxygen dismustase (SOD) concentrations after 15 days.ResultsIn the 6 h HBO,24 h HBO,48 h HBO and 72 h HBO groups,average neuron density in the CA1 region was significantly higher than in the HI group.But in the 1 week HBO group the average density was not significantly different than in the HI group.In the 6 h HBO,24 h HBO,48 h HBO and 72 h HBO groups the average percentage of neuron cell apoptosis in the CA1 area of the hippocampus was significantly lower than in the HI group,but the 1 week HBO group again showed no significant difference.There were significant differences in average NO,MDA or SOD levels among the 6 h HBO,24 h HBO,48 h HBO,72 h HBO and HI groups,but the 1 week HBO group showed no significantly higher average levels than the HI group.ConclusionsThe optimal therapeutic window for using HBO to protect against HIBD is within the first week.The best effect can be obtained in the first 6 hours,but after 1 week HBO no longer has a significant effect.The earlier the HBO is administered,the better the effect obtained.

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