Objective To investigate the mechanism of endothelin (ET) in acute biliary pancretitis (ABP)with renal impairment, and examine the improvement of renal function by puerarin. Methods Thirty-two patients of ABP with renal impairment were selected and were randomly divided into group A and group B. Group A were given puerarin post-operatively, while group B were given common manegement postoperatively. Meanwhile, 16 ABP patients without renal impairment were selected as group C, and 16 nonABP patients who had common hepatic or biliary diseases were selected as group D. The levels of plsma ETand creatinine clearance rate(Ccr) of renal function in each group were observed pre-operation and 1 week post operation. Results In pre-operation the ET level in group A, B and C were higher than in group D(P ＜0.001), and the ET level in group A,B was higher than in group C(P＜0.001). Post operation after I week the ET level in group A was lower than in group B(P =0.014), while the Ccr level in group A was higher thanin group B ( P = 0.002). Conclusions The function of endothelin is an important mechanism in biliary pancretitis with renal impairment. Puerarin might reduce the ET level and increase the Ccr level in patients of ABP with renal impairment, improve the renal function of such patients.

BACKGROUND: Previous results of our study show that oyster extract has some protective effects on apoptosis of the neuroepithelium in neural tube defects induced by hyperthermia in vivo.It remains unclear whether the extraet also protects in vitro cultured neural stem cells.OBJECTIVE: To investigate the protective effect of oyster extract on apoptosis of cerebral neural stem cells induced by hyperthermia.METHODS: The cerebral neural stem cells of embryonic mice of 13 days were cultured in vitro.Nestin expression was detected by immunofluorescence method to identify neural stem cells.The neural stem cells of passage 3 were divided randomly into 4groups: hyperthermia control group and oyster treated Ⅰ,Ⅱ,Ⅲ groups(mass concentration 2.5,5,10 g/L oyster extract solution).In addition,culture solution control group(no cells),and culture solution+oyster extract control group(no cells)were designed.All oyster extract groups and control groups were treated by hyperthermia over 39 ℃.The survival rate and the vitality of neural stem cells were detected by trypan blue staining and MTT assay.Western-blotting was employed to explore the expression of p53 in cerebral neural stem cells of each group.RESULTS AND CONCLUSION: The survival rate and the value of MTT assay in oyster treated groups Ⅱ and Ⅲ were significantly greater than hyperthermia control group(P < 0.05),but the expression of p53 in oyster treated groups Ⅱ and Ⅲ were weaker than hyperthermia control group.Oyster extract plays an important protective role in the apoptosis of neural stem cells induced by hyperthermia.

Objective To observe the characteristics of cognitive impairment in patients with primary hypothyroidism.Methods A total of 90 primary hypothyroidism patients untreated with thyroid hormone were selected.All the 90 patients were divided into subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group,and 30 patients in each group.The other 30 healthy volunteers were selected as controls.Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used for the testing their orientation,immediate memory,attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power.One-way analysis of variance was performed to determine significant differences among four groups.Results The MMSE scores in subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group((27.53 ± 2.16),(26.90±1.88) and (24.80 ± 2.10) respectively) were lower than those of the control (28.23 ± 1.33).The MoCA scores of the above hypothyroidism groups ((23.57 ± 3.33),(2 1.60 ± 2.81) and (20.53 ± 3.03) respectively) were also lower than that of the control (26.63 ± 2.31) (P ＜ 0.05).Except for orientation and immediate memory,statistical significances of the other cognitive function were existed between hypothyroidism groups and the healthy controls (P＜ 0.05).With the increase in severity of hypothyroidism,the abnormality of attention,calculation,linguistic competence,visual space and executive ability,naming ability and abstracting power were appearing gradually in hypothyroidism groups (P ＜ 0.05),and the scores were low(P＜ 0.05).Conclusion Defects of attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power are existed in primary hypothyroidism patients.

Objective To investigate the changes and significances of peripheral blood CD34,KDR/CD34,CD133/CD34,and CD117/CD34 positive cell levels in patients with hemorrhagic stroke at the acute phase. Methods Thirty patients with acute hemorrhagic stroke admitted to the Department of Neurosurgery, Jiangsu Haimen People’s Hospital from September 2013 to April 2014 were enrolled retrospectively. At the same time,20 healthy subjects were selected as a control group. CD34 +,KDR+ /CD34 +,CD133 + /CD34 +, and CD117 + /CD34 +cell levels in peripheral blood were detected in patients with hemorrhagic stroke at day 1 to day 7 after cerebral hemorrhage and the day of physical examination of the control group using FACSCalibur flow cytometry. The data were obtained and analyzed using CELLQuest software. Results Peripheral blood CD34 + cells and KDR+ /CD34 +, CD133 + /CD34 +, CD117 + /CD34 + cells at day 1 to day 7 after ischemic stroke were all lower than control group (all P<0. 05). CD34 + and CD117 + /CD34 + cells decreased firstly at day 1 and 2, then increased gradually;KDR+ /CD34 + and CD133 + /CD34 + cells increased gradually at day 1 to day 5,they were all reached the peak at day 5,which were (14. 8±3. 5) í105 and (16. 7±3. 3) í105 respectively. Compared with that at day 1,CD34 + cells at day 5 and 6 were (27. 4 ±6. 3) í105 and (25. 4 ±5. 7) í105 respectively;KDR+ /CD34 + and CD133 + /CD34 + cells at day 4,5,and 6 were (10. 2±3. 1) í105,(14. 8±3. 5) í105,(12. 1±3. 4) í105 and (14. 3±3. 6) í105,(16. 7±3. 3) í105,and (13. 1±4. 0) í105,respectively;CD117 + /CD34 + cells at day 5 was (21. 3 ±4. 2) í105,which were all higher than the first day after cerebral hemorrhage (P<0.05). In the CD34 + cells,the proportion of KDR+ /CD34 + cells at day 1 to day 7 all decreased compared with the control group (all P<0. 05);the proportion of CD133 + /CD34 + cells at day 4 was (65±4)%,it was higher than the control group (P<0. 05);the proportion of KDR+ /CD34 + cells at day 5 was (55±6)% compared with day 1;the proportion of CD133 + /CD34 + at day 4 was (65 ±4)%;the proportions of CD117 + /CD34 + cells at day 4 and day 5 were (69 ±6)% and (72 ±6)% respectively,and they all increased (all P <0. 05). Conclusion Peripheral blood CD34 + cells and their cell subsets are change in patients with hemorrhagic stroke in acute phase. Speculating the different cell subsets may have different functions and potential. KDR+ /CD34 + and CD133 + /CD34 + may be the early sensitive indicators of acute hemorrhagic stroke,and CD117 + /CD34 + is largely mobilized in early stage.

The paper is aimed to study the difference in yield and quality at different harvest time and determine the optimum harvest of planting Gentiana in Ludian traditional harvest period. The authors analyzed the variation in fresh weight, dry weight, dry discount rate, length, diameter, volume and the content of gentiopicroside, loganin acid, alcohol-soluble extract and total ash and made a comprehensive appraisal of yield, appearance quality and intrinsic quality by gray relational distance ideal Comprehensive Evaluation method. The results showed that there is a big difference in yield and quality both 2-year-old and 3-year-old Gentiana harvested in traditional harvest period and the comprehensive evaluation more better when harvested more later. It can be seen, Gentiana harvested the later had a better yield and quality in Ludian traditional harvest period. The harvest of Gentiana can be appropriate delayed depending on the particular circumstances of production.
Agriculture ; methods ; China ; Gentiana ; anatomy & histology ; growth & development ; metabolism ; Iridoid Glucosides ; metabolism ; Organ Size ; Quality Control ; Time Factors

Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals ; Base Sequence ; Cloning, Organism ; Electrophoresis ; Endonucleases ; genetics ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; Gene Targeting ; methods ; Goats ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Zinc Fingers

Anemia, Aplastic ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans

BACKGROUND: The existence of minimal residual leukemia cells is the main cause for the recurrence of acute leukemia in children, and immunological biological therapy has attracted more and more attentions in the various methods from eliminating minimal residual disease. Previous studies have found that interferon α-2b can effectively inhibit the increase of tumor cells in vivo in children with neuroblastoma and malignant lymphoma, whether it can inhibit the increase of leukemia cells?OBJECTIVE: To investigate the effects of interferon α-2b in vitro on leukemia cells.DESIGN: A comparative observation taking human promyelocytic leukemia HL-60 cell line as the material.SETTING: Cell Culture Room; Immunological Laboratory; Cell Room, Institute of Pediatrics, Affiliated Hospital,Medical College of Qingdao University.MATERIALS: HL-60 cell line was provided by Shandong Institute of Basic Medical Sciences. Interferon α-2b was purchased from Megagene Company Fluorescein isothiocyanate (FTTC) rabbit-anti-rat Ig solution (CatEK001) and CD13 anti-human monoclonal antibody solution (Cat. DK013Y) were purchased from Union Stem Cell & Gene Engineering Co.,Ltd.METHODS: The experiments were carried out in the Institute of Pediatrics, Affiliated Hospital, Medical College of Qingdao University from March to September 2005. HL-60 cells culture system was established in vitro, and the oncentration was adjusted to 1×109 L-1. The cells were divided into control group and experimental group. In the experimental group, each well was added by interferon-α-2b with the terminal concentration of 5×105, 1×106, 2×106,5×106 and 1×107 U/L, respectively. In the control group, each well was added by saline of the same volume. The cells were cultured continuously for 48 hours. The morphological changes of HL-60 cells were observed using Wright's staining under light microscope; Cell apoptosis was observed using acridine orange/ethidium bromide double staining; Antigen expression and maturation and differentiation on cell membrane were observed by determining CD13 protein expression; Proliferation and activity of HL-60 were detected with methyl-thiazol-tetrazolium (MTT) assay.MAIN OUTCOME MEASURES: The occurrence of apoptosis was judged according to the uniformity and staining of HL-60 nuclear chromatin; HL-60 cell proliferation was judged according to the absorbance (A) value; The maturation of HL-60 cells was judged according to the number of positive CD13 cells.RESULTS: ① HL-60 cell apoptosis: The cells were cultured for 48 hours. When the concentration of interferon α-2b was 5×105 U/L, there were mainly early apoptotic HL-60 cells; When the concentration was 1×107 U/L, there were mainly late apoptotic cells, and the apoptotic rate was significantly higher than those in the control group (P ＜ 0.01 ).② HL-60 cell proliferation: The A values in the experimental groups treated with interferon α-2b of 2×106 U/L and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01). ③ Maturation of HL-60 cells: The percentages of positive CD13 cells in the experimental groups treated with interferon α-2b of 1 ×106 and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01).CONCLUSION: It is concluded that interferon α-2b can enhance the apoptosis, inhibit the proliferation and promote maturation and differentiation of HL-60 cells.

Objective:To study the effect of chemokines CCL20 and CCL22 combined with skin-induced Treg on survival time of grafted skin.Methods: Skin grafting mice were divided into four groups, three mice per group, namely Treg group, Treg+CCL20 group,Treg+CCL22 group and control group.C57BL/6 mice were used as donor and BALB/c as acceptor, and the Treg cells were isolated from the mice induced by skin allograft.After skin grafted,CCL20 and CCL22 were subcutaneous injection every day,which lasted for 10 day.Survival time of skin in each group were observed and recorded.The Treg colonzation experiments were performed as follows.We firstly isolated Treg with Magnetic cell sorting system( MACS) and then labled them with 99 Tcm.After that we intravenously injected them into the mice.3 hours later,the mice were sacrifced and the radioactivity of organs were detected by GC-2016γradioim-munoassay counter.Results:①After Treg treated the survival time of skin grafted in antigen-induced Treg group was signifiantly longer than control group,when treg were cooperated with CCL20 and CCL22,the skin grafted showed more longer survival time than Treg and control groups( P<0.001 ).②After injection of induced Treg, Treg in autologous and allogeneic skin grafts goups were mainly distributed in autologous and allogeneic skin,accounting for 60% and 98% respectively.When cooperated with CCL20 or CCL22,the Treg were mainly distributed in liver.Conclusion:Chemokines CCL20 and CCL22 synergistically improved the effects of skin antigen induced Treg on survival time of skin graft,which probably related with the Treg colonization into the liver.

Objective To analyze the expression of the placenta Grb10 from women conceived by transferred thawed blastocyst,and to evaluate the security of blastocysts vitrification.Methods A cross-sectional study was performed in the Department of Obstetrics and Gynecology of Fujian Provincial Maternity and Children's Hospital from January 2012 to May 2014,50 women conceived by transferring thawing blastocyst and 50 natural pregnancy control women were enrolled in this study.The expression of Grb10 protein was detected by immunohistochemistry and Western blot,and the expression of Grb10 mRNA was detected by Realtime PCR method.Results Comparison of two cases of gestational age,gestational age,fetal sex,fetal body weight,body length,head circumference,abdominal circumference,there were no significant differences(P＞0.05),comparison of placental area,placental weight,the difference was statistically significant(P＜0.05).Real-time PCR and Western blot results showed that,there was no significant difference in the expression of Grb10 mRNA and protein between the two groups(P＞0.05).Conclusion Blastocysts vitrification may increase the area and quality of delivery of placenta,however,there was no significant change in the expression of Grb10 in placenta.