Objective To clone the murine ?-fetoprotein gene,construct the DNA vaccine of mouse AFP and to observe the specific cellular immunologic responses and the anti-tumor responses in mice induced by this vaccine.Methods The murine AFP gene was amplified by RT-PCR from total RNA extracted from Hepa1-6 cells and cloned into the vector pcDNA3.1 to construct pmAFP.The DNA vaccine was identified by restriction enzyme analysis,sequencing and expression.EL-4(mAFP) was developed by stable transfection of EL-4 cells with pmAFP.Mice immunized with DNA vaccine by intramuscular injection were inoculated with EL-4(mAFP) cells at back to observe the inhibitory effect of the immunization on tumor.The frequency of cells producing IFN-? in splenocytes harvested from the immunized mice was measured by ELISPOT.Blood samples were collected from the immunized mice to check the functions of liver and kidney.Results The murine AFP gene was successfully cloned by RT-PCR.DNA vaccine was successfully constructed.The expression of mAFP mRNA in EL-4(mAFP) was confirmed by RT-PCR.The tumor volume in pmAFP vaccine group(1 042.42?123.71 mm~(3)) was significantly smaller than that of other groups(P

To clone the murine alpha-fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1-6 cells, and then the murine alpha-fetoprotein gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant of vector was identified by restriction enzyme analysis and sequencing. After transient transfection of CHO cells with the vector, Western blotting was used to detect the expression of AFP. It is concluded that the 1.8 kb murine alpha-fetoprotein gene was successfully cloned and its eukaryotic expression vector was successfully constructed.
CHO Cells ; Cloning, Molecular ; Cricetinae ; DNA, Complementary ; Eukaryotic Cells/metabolism ; Genetic Vectors ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; alpha-Fetoproteins/*biosynthesis ; alpha-Fetoproteins/genetics

Rhodiola capsules show the resistant effects on ischemia, anoxia, radiation, fatigue and virus and so on, and can im-prove the body immune function to relief the plateau symptoms such as fatigue and cerebral hypoxia. The influence mechanism of rhodi-ola capsules in the effect of anti-high altitude pulmonary edema was summarized in the paper.

The main feature of acute steroid-resistant rejection (ASRR) after liver transplantation is a lack of response to steroids.Although some effective immunosuppressants are available,there is no uniform treatment presently available.This paper will focus on exploring the mechanism of ASRR and formulating individually approached strategies.

BACKGROUND:There are more ways of internal fixation in treatment of elderly intertrochanteric fracture;however, how to choose a fixed way is the key issue of ensuring the efficacy. OBJECTIVE:To compare the therapeutic effect of proximal femoral anti-rotation intramedul ary nail and proximal femoral locking plate in the treatment of elderly femoral intertrochanteric fractures. METHODS:Total y 100 patients with femoral intertrochanteric fractures who received the treatment at Qionghai City People’s Hospital from December 2009 to December 2013 were randomly divided into locking plate and anti-rotation intramedul ary nail groups. Patients in the anti-rotation intramedul ary nail group underwent internal fixation using proximal femoral anti-rotation intramedul ary nail. Patients in the locking plate group underwent internal fixation using proximal femoral locking plate. RESULTS AND CONCLUSION:Compared with the locking plate group, the amount of bleeding, operation time, postoperative bed activity time and hospital stay in the anti-rotation intramedul ary nail group significantly reduced, the excel ent rate of Harris scores was significantly increased after 1 year of internal fixation and the incidence of complications and adverse reactions was significantly decreased. There was no host response to the material in these two groups.

Objective To explore the effect of beraprost sodium (BPS) on hypoxia-induced pulmonary artery hypertension (HPH) in rats and the expression of oxygen-sensitive Kv channels in pulmonary artery smooth muscle (PASM).Methods The HPH model of rats was established by exposing rats to low-pressure and low-oxygen cabin which was auto-modulated for 8h every day.Rats in the BPS group were given an intragastric administration of BPS [300 μg/(kg · d)],while those in the control group and HPH group were given an intragastric administration of 3 ml/kg of 0.9％ saline.After 4 weeks,the mean pulmonary artery pressure (mPAP) was measured and right heart ventricle hypertrophy index (RVHI) was calculated;pulmonary artery remodeling was evaluated by HE staining;the expressions of Kv 1.2,Kv 1.5 and Kv2.1 in the pulmonary artery were detected by Real-time PCR and Western blot.Results The HPH model was successfully established in rats exposed to chronic hypoxia for 4 weeks.Compared with those in HPH group,mPAP,RVHI and pulmonary artery remodeling were decreased in BPS group [mPAP:(13.48±2.18)mmHg vs.(23.87±2.23)mmHg vs.(17.09±1.20)mmHg;RVHI:0.28±0.02 vs.0.46±0.03 vs.0.36±0.04;％ area of medial smooth muscle:35.72±6.58 vs.68.52±5.64 vs.46.58±8.43;P＜0.05],and the mRNA and protein expressions of Kv 1.2,Kv 1.5 and Kv 2.1 were increased (relative protein expression level:Kv1.2,0.78±0.10 vs.0.15±0.03 vs.0.57±0.13;Kv1.5,0.61±0.10 vs.0.31±0.05 vs.0.59±0.13;Kv2.1,0.29±0.05 vs.0.10±0.02 vs.0.28±0.07;P＜0.05).Conclusion BPS can improve pulmonary arterial hypertension induced by hypoxia,and upregulate the decreased mRNA and protein expressions of Kv 1.2,Kv 1.5 and Kv 2.1 in pulmonary artery.

AIM:To investigate the expression of poly(ADP-ribose) polymerase-2 (PARP-2) during rat car-diac hypertrophy in vitro and in vivo, and to explore the effects of PARP-2 on the cardiac hypertrophy.METHODS:Ab-dominal aortic coarctation ( AAC) was performed to establish a model of pressure overload-induced cardiac hypertrophy in SD rats.The expression of PARP-2 at mRNA and protein levels in the myocardium was determined by real-time PCR and Western blot.The hypertrophy model of the cardiomyocytes was induced by treating the cells with angiotensinⅡ( AngⅡ) . PARP-2 was knocked down by siRNAs in neonatal rat cardiomyocytes and the cardiomyocyte hypertrophy was evaluated by measuring the mRNA levels of ANF, BNP, and β-MHC and the cellular surface area.RESULTS: The expression of PARP-2 at mRNA and protein levels was both increased in the AAC rats as compared with those in the sham animals.The expression of PARP-2 at mRNA and protein levels was also increased in a time-and concentration-dependent manner in AngⅡ-induced hypertrophy model of the cardiomyocytes.In the neonatal rat cardiomyocytes, knockdown of PARP-2 ex-pression by siRNA attenuated AngⅡ-induced cardiac hypertrophy of the cardiomyocytes, indicating that endogenous PARP-2 played a positive regulatory role in cardiac hypertrophy.CONCLUSION:The mRNA and protein levels of PARP-2 in-crease in the in vitro and in vivo models of cardiac hypertrophy.Knockdown of PARP-2 protects cardiomyocytes from hyper-trophy.

Female ; Head and Neck Neoplasms ; pathology ; Humans ; Infant ; Teratoma ; pathology

Objective To explore the neuroprotective effect of repeated preconditioning with cannabinoid receptor agonist WIN 55,212-2 on focal cerebral ischemia-reperfusion injury in rats.Methods Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO)for 120min.Fifty male SD rats were randomly assigned to five groups(10 each):rats in control group and dimethyl sulphoxide group(DMSO group)were intraperitoneally administered 0.3ml normal saline and 0.3ml DMSO once a day for 5 days.Rats in WIN 55,212-2 preconditioning groups(including WIN1,WIN3 and WIN5 group)received intraperitoneal injection of 1mg/kg WIN 55,212-2(dissolved with 0.3ml DMSO)once a day for 1d,3d and 5d,respectively.All animals underwent MCAO operation 24h after last pretreatment to reproduce temporal(120min)focal cerebral ischemia model.The neurological function score(NFS)was evaluated at 24,48 and 72h after reperfusion.Brain infarct was identified with 2% 2,3,5-triphenyltetrazolium chloride(TTC)staining 72h after reperfusion,and the brain infarct volume was expressed as percentage of normal cerebral hemisphere volume.Results The NFSs of rats in WIN 55,212-2 preconditioning groups(WIN1,WIN3 and WIN5 group)were significantly higher,and the infarct volumes were significantly smaller than that in control group and DMSO group at 24h,48h and 72h after reperfusion(P0.05).Conclusion The neuroprotective effect of repeated preconditioning with cannabinoid receptor agonist WIN 55,212-2 may be enhanced by increased time of pretreatment.

Objective To investigate the correlation of vitamin D with essential hypertension and its target organ damages in the elderly.Methods In the retrospective study,365 elderly participants aged ≥ 60 years hospitalized in our department during January 2013 to February 2014 were involved,including 247 patients with confirmed essential hypertension(HT)and 118 patients without HT.Serum 25-hydroxy vitamin D[25(OH)D]levels were measured and compared between HT and non-HT group,and binary regression analysis was used to discuss potential influencing factors of HT.The damage index of vitamin D target organ was compared between vitamin D deficiency group(n=198)and vitamin D non deficiency group(n=49).Results The median serum 25(OH)D level of participants is 35.3 nmol/L,77.5%(283 cases)were in the insufficient vitamin D status.The level of 25(OH)D was significantly lower in HT group than in non-HT group(34.0 vs.38.7 nmol/L,P<0.05).There were not statistically significant differences between vitamin D insufficient group and sufficient group(P>0.05)in albumin/creatinine ratio,serum creatinine,estimated glomerular filtration rate,ankle-brachial index,pulse wave velocity,intima media thickness of carotid artery,Sokolow-Lyon index,cornell index and left ventricular mass index.The serum 25(OH)D level was significantly lower in CHD plus HT group than non-CHD(P<0.05).The 25(OH)D level was negatively correlated with coronary heart disease(B=-0.021,P=0.019).The serum 25(OH)D level was significantly lower in groups with double-and triple-vessel coronary artery disease than in non-CHD group(P<0.01).Conclusions Vitamin D insufficiency is common in the elderly.The serum 25(OH)D level is significantly lower in old patients with HT especially with stage 3 HT than in non-HT ones.The 25(OH)D level is lower in HT patients with comorbid CHD,and more obvious in HT patients with multi-vessel coronary artery disease.An insufficient vitamin D status is not correlated with hypertensive target organ damages including kidney,peripheral vascular and left ventricular cardiac structure.