Objective To preliminary investigate the cytoactive of HELA cell after PUMA gene promoting HELA cell apoptosis and whether or not the protein of cyt-c,AIF in the chondrosome have participated in the process of PUMA gene inducing HELA cell.Methods we Used AO/EB dyeing to detect the Morphologic change of cell induces by PUMA gene and Western Blot to detect whether or not the position of cyt-c,AIF in the chondrosome have transferred after PUMA gene induces HELA cell apoptosis.Result ①Fluorescence microscope and inverted phase contrast microscope display HELA cell have a series of apoptosis Morphologic change after transfecting PUMA into HELA cell,and this Morphologic change of cell is more obviously after transfecting 48h than after 24h.;②Western Blot display the protein cyt-c and AIF transferred toward to kytoplasm after HELE cell took place apoptosis,the mount of transfecting protein is more obviously after transfecting 48h than after 24h.Conclusion ① PUMA gene have the effect of promoting HELA cell apoptosis;②cyt-c and AIF take part in the process of promoting HELA cell apoptosis,but during this process if the membrane potential of chondrosome was changing need to do some more study to confirm it.

Objective: To express recombinant human angiostatin for further application in clinic. Methods: The complete encoding eDNA of human angiostatin was isolated from human embryo liver with RT-PCR and expressed in secretory Pichia expression system. Recombinant human angiostatin was purified with heparin sepharose chromatography and its activity was determined in chick embryo chorioallantoic membrane (CAM) and wound healing assays. Results: Expressed in large quantity (yield=5 mg/L) and purified with heparin sepharose, recombinant angiostatin was showed to have a molecular weight of 43 kD in SDS-PAGE and potently inhibit angiogenesis and wound healing. Conclusion: Recombinant human angiostatin was expressed efficiently in a biologically active form.

Abstract Objective:To construct bmcella melitensis Br.melitensis genomic DNA expression library and study its immune effect.Meth-ods: Hind Ⅲ digested Br.melitensis genomic DNA fragments were cloned into pSV-?-Gal plasmds.The recombinant plasmids were transformedinto JM109 host bacteria. After large-scale isolation, the recombinant plasmids were inoculated into the quadriceps muscle of mice. In the 9thday after third boost immunization .agglutination antibodies against Br. melitensis and lymphocyte mitogenic ability of mice were determined. Re-sults : Expression library nucleic acid vaccine can induce the production of agglutination antibodies against Br. melitensis and stimulate the lym-phocyte porliferation primed by ConA. Conclusion:The genomic DNA expression library may provide a new vaccine against Br. melitensis infec-tion.

Objective:To study the clinical application potential of aFGF and try to produce aFGF as gene engineering medicine.Methods:The complete encoding cDNA of human aFGF was isolated from human lung fibroblast with RT PCR.Recombinant human aFGF was expressed in secretory pichia expression system and purified with heparin sepharose chromatography.The activity of aFGF was detected in NIH3T3 proliferation assay,chick embryo chorioallantoic membrane assay(CAM) and wound healing assay.Results:The recombinant aFGF was expressed in large quantity(yield=12 mg/L) and was capable to promote proliferation of NIH3T3,angiogenesis and wound healing.Furthermore,those activities of aFGF could be agonized by recombinant FGFR extracellular domain.Conclusion:Recombinant human aFGF was expressed efficiently and possessed natural biological activities.

Objective To determine the effect of high dose albumin on permeability of blood brain barrier (BBB) in brain of rats after ischemic-reperfusion (IR) in order to explore its possible mechanism.Methods Establishment of brain ischemic reperfusion rat model by using middle cerebral artery occlusion (MCAO).Medicine treatment was given by caudal vein injection after 2 hours of MCAO.Thirty-six healthy male SD rats were then randomly (random number) divided into 6 groups (n =6 in each):6 h and 24 h sham-operation groups (Group Sham:operation without ischemia),6 h and 24 h normal saline groups (Group NS:NS injection 5 ml/kg) and 6 h and 24 h albumin group (Group Alb:25 ％ Alb injection 1.25 g/kg).Six hours and 24 hours after the end of reperfusion,rats were measured by Zea-Longa score (neural function deficit) separately.Serum concentration of S100B was examined by the ELISA kit and Evans blue in brain tissue was detected by spectrophotometer.The level of AQP4 was examined by Western blot and immunohistochemistry.All data were analyzed by one-way analysis of variance (ANOVA),The intergroup comparisons were analyzed by the least-significant-difference (LSD) test by using SPSS version 17.0 software.Differences were considered statistically significant if P ＜ 0.05.Results Zea-Longa score significantly increased in both group NS and group Alb at 6 h and 24 h (P =0.000).However,there was no significant difference in ZEA-LONGA score of 6 h and 24 h between group Alb and group NS (P =1.000).The serum concentration of S100B in group NS 6 h was significantly lower than that in group Alb at 6h (196.67±20.11 vs 160.04±14.00,P=0.000),and at24h (2.45±0.07 vs.2.23±0.07,P=0.000).Furthermore,concentration of Evans blue in brain tissue in group Alb was significantly higher than that in group NS at both 6 h (0.97 ± 0.08 vs.0.74 ± 0.06,P =0.000) and 24 h (2.45 ± 0.07 vs.2.23 ± 0.07,P =0.000).The expression of AQP4 in brain tissue was higher in group Alb than that in group NS at both 6 h (0.72 ±.0.11 vs.0.57 ± 0.06,P ＜ 0.01) and 24 h (0.80 ± 0.03 vs 0.61 ± 0.02,P ＜0.01).Conclusions High dose albumin contribute slightly in improvement of neural deficit in rats after IR.On the contrary,it can also aggravate the IR injury,which increases brain edema then increase the permeability of BBB.The mechanism may be associated with over-expression of AQP4 in brain tissue.

Objective To investigate regeneration and repair effect after ChABC,GDNF and Nogo-A Ab combination treatment for experimental spinal cord injury model.Methods Rat (T7-8 )complete spinal cord injury crosscutting animal model was established.The SD rats were randomly divided into 6 groups:normal group, sham operation group,simple transection group,A (ChABC)group,G (GDNF)group,N (Nogo-A antibody) group,and AGN (ChABC+GDNF+Nogo-A antibody)group.At 24 w after spinal cord injury,BDA tracer,NF-200,GAP-43,and GFAP immunohistochemistry were evaluated.Results BDA tracer of A group,G group and N group showed dye light,the proximal end of damaged zone showed the blue tracer particles,while damaged zone showed few blue regenerated nerve fibers.AGN group showed visible blue nerve fibers through the damaged zone and the distal segment in the damaged zone;the central zone of injury vacuolar degeneration showed the blue dyed fibers.NF-200 immunohistochemical staining showed NF-positive staining in A group,AGN was stronger than that in control group and simple transection group (P <0.05),AGN group than G group,N group significantly increased (P <0.05).GAP-43 positive staining in A group and AGN treatment group was stronger than in control group and simple transection group (P <0.05),AGN group significantly increased compared with A,G and N groups (P <0.05).GFAP positive staining in control group and simple transection group was stronger than in each treatment group (P <0.05),but A group,G group,N group and AGN group showed no significant differences (P > 0.05 ).SEP wave was detected in control group and AGN group,while the latency time was longer in AGN group than in control group.Conclusion ChABC,GDNF,and anti-Nogo-A antibody used alone or in combination can improve spinal cord injury and nerve cell function,and the joint application could improve regeneration after spinal cord injury than any monotherapy.

The relationship between the expression of resistin in polycystic ovary syndrome (PCOS) and insulin resistance was investigated. The plasma resistin concentrations in 35 patients with PCOS and 40 controls were measured by ELISA. Luteinizing hormone (LH), follicle-stimulating hormone (FSH), and fasting insulin (FIN) were tested by radioimmunoassay. Insulin resistance index (HOMA-IR) was calculated. Fasting plasma glucose (FPG) was determined by oxidase test. Western blot and reverse transcriptase PCR (RT-PCR) methods were used to detect the expression of resistin in adipose tissues. The levels of plasma resistin, LH, LH/FSH and FIN and HOMA-IR in patients with PCOS were significantly higher than those in control group (all P<0.05). Plasma resistin was correlated positively with FPG, FIN, HOMA-IR, LH and LH/FSH (r=0.56, 0.60, 0.65, 0.48, and 0.42 respectively). Resistin protein and mRNA expression levels in patients with PCOS were significantly higher than those in normal tissues (all P<0.01). It was concluded that resistin might be involved in the pathogenesis of insulin resistance of PCOS.

BACKGROUND:Neural stem cel transplantation provides an important way to treat sequela of traumatic brain injury, but the timing for treatment is inconclusive.
OBJECTIVE:To explore the clinical effect of neural stem cel transplantation in the treatment of sequela of traumatic brain injury and the choice of the best treatment time.
METHODS: Totaly 178 patients with sequela of traumatic brain injury who underwent neural stem cel transplantation were divided into three groups as per the timing for neural stem cel transplantation: group A (with 6 months after injury,n=60), group B (6-12 months after injury,n=59), and group C (over 12 months after injury,n=59). Improvement in clinical symptoms and scores on function independent measure (FIM) were recorded and compared in the three groups.
RESULTS AND CONCLUSION:The total effective rate of group A was significantly higher than that in groups B and C (P < 0.05). FIM scores were significantly improved in the three groups after cel transplantation (P < 0.05). At 3 months after the fourth transplantation, the FIM score in the group A was significantly higher than that in the other two groups, and the incidence of adverse reactions in the group A was significantly lower than that in the other two groups (P < 0.05). These findings indicate that neural stem cel transplantation at different timing can al harvest certain clinical effects, but the best timing for neural stem cel transplantation is within 6 months after injury.

Objective To investigate the usage of antiepileptic drugs (AEDs),epileptic seizures,pregnancy outcomes and infant feeding practices among pregnant women with epilepsy,and provide scientific evidence for eugenics.Methods The clinical characteristics of 350 pregnant women with epilepsy from Shaanxi Provincial Epilepsy & Pregnancy Register from October 2012 to July 2016 were analyzed retrospectively.Results A total of 350 patients with 376 pregnancies were included in the final analysis.Among 376 pregnancies,272 pregnancies (72.3％) were under the treatment of AEDs.Of them,246 pregnancies (90.4％) were on monotherapy,26 (9.6％) on polytherapy.Only patients during 179 pregnancies (47.6％) took folic acid in the first trimester.Seizure frequency increased in patients during 123 pregnancies (32.7％),of whom patients during 43 pregnancies (35.0％) did not use AEDs.Although other patients during 80 pregnancies (65.0％) took AEDs,regular pharmacokinetic monitoring was absent.A total of 170 pregnancies (45.2％) attained seizure-free.Seizure frequency decreased in 42 pregnancies (11.2％) and remained unchanged in 24 pregnancies (6.4％).The rates of cesarean section and natural delivery were 43.8％ (158/361) and 56.2％ (203/361),respectively.Totally 4.5％ (17/376) pregnancies had adverse pregnancy outcome,and 1.9％ (7/376) had fetal malformations.The most common congenital malformation was heart malformation,which was observed in two offspring.One offspring developed dysostosis,one developed chromosome abnormality,and another developed cerebellum deformity.In addition,one twin developed conjoined deformities.Breastfeeding rate was 52.6％ (190/361).Conclusions In Shaanxi province,AEDs compliance,pharmacokinetic monitoring and usage rate of folic acid still need to be improved in pregnant women with epilepsy.Clinical trial registration Chinese Clinical Trial Registry,ChiCTR-OOC-16009277

Foam Cells ; pathology ; Gastrointestinal Neoplasms ; pathology ; Humans