Objective: To construct the prokaryotic expression vector bearing fusion gene NT4-ADNF-9 and lay foundation for further study on genetic therapy of neurosensory deafness. Methods: By means of asymmetrical primer/ template, double stranded cDNA of activity dependent neurotrophic factor-9 (ADNF-9) was obtained, which included restriction enzymes sites on the two extremities. ADNF-9 cDNA was ligated to the signal and leader peptides of neurotrophin 4 (NT4), and the fusion gene was named NT4-ADNF-9. Then it was subcloned into prokaryotic expression vector pBV220, and called pBV220/ NT4-ADNF-9. Results: Evidences of DNA sequence analysis and restriction enzymes digestion showed that we recombined ADNF-9 cDNA to the 3′terminal of the signal and leader peptides of NT4, and the fusion gene was subcloned into pBV220 successfully. Bioactivity of the products was proved that it could support the cell survival and neurite growth in the primary cultures of dorsal root ganglia (DRG) of embryonic day-8 chicken neurons as compared to the control. Conclusion: Prokaryotic expression vector pBV220/NT4-ADNF-9 can be constructed successfully and the bioactivity is satisfactory.

Objective: To construct the prokaryotic expression vector bearing fusion gene NT4-ADNF-9 and lay foundation for further study on genetic therapy of neurosensory deafness. Methods: By means of asymmetrical primer/ template, double stranded cDNA of activity dependent neurotrophic factor-9 (ADNF-9) was obtained, which included restriction enzymes sites on the two extremities. ADNF-9 cDNA was ligated to the signal and leader peptides of neurotrophin 4 (NT4), and the fusion gene was named NT4-ADNF-9. Then it was subcloned into prokaryotic expression vector pBV220, and called pBV220/ NT4-ADNF-9. Results: Evidences of DNA sequence analysis and restriction enzymes digestion showed that we recombined ADNF-9 cDNA to the 3′terminal of the signal and leader peptides of NT4, and the fusion gene was subcloned into pBV220 successfully. Bioactivity of the products was proved that it could support the cell survival and neurite growth in the primary cultures of dorsal root ganglia (DRG) of embryonic day-8 chicken neurons as compared to the control. Conclusion: Prokaryotic expression vector pBV220/NT4-ADNF-9 can be constructed successfully and the bioactivity is satisfactory.

Objective To evaluate the preoperative nutritional status of patients with gastric carcinoma by using the European Nutritional Risk Screening 2002(NRS 2002)and its prediction for postoperative nutrition-related complications.Methods We prospectively evaluated the nutritional risk of 314 gastric cancer patients admitted in one center from 2004 to 2007 with NRS 2002 with China's normal body mass index(BMI),in terms of postoperative complications,mortality and hospital stay.Results NRS 2002 scoring system was applicable in 93.1% cases.Preoperatively 125 patients were of score≥3,accounting for 39.8% of this group.The postoperative complication rate(26.2%)was higher than 13.8% in those with normal preoperative nutritional scores(NRS 2002 score＜3)(P＜0.05);The odds ratio to develop a complication was 0.642 in patients with preoperative nutritional risk score(P＜0.05),and 1.596 in patients with clinicopathological stage of gastric cancer(P＜0.01).The correlation between length of hospital stay and nutritional risk was also assessed by Pearson correlation analysis.The Pearson coefficient was 0.177(P=0.002).Conclusion Preoperative nutrition score(NRS 2002)≥3 predicts higher postoperative complications and longer hospital stay.Preoperative nutritional support is necessary in patients with preoperative nutrition score(NRS 2002)≥3.

Objective To explore the cosmetic effect of oncoplastic volume displacement in breastconserving surgery.Methods 43 cases of breast cancer in stages I and Ⅱ with pathological diagnosis by core needle biopsy were operated by extended resection of tumor combined with axillary lymph nodes dissection.Among the patients,wound cavity was repaired by direct skin suture without suture of mammary gland deletion in 22 cases,and another 21 cases were repaired by oncoplastic volume displacement technique in which skin suture followed by freeing remaining breast tissue flap and translocation to repair the mammary defect.All of them were treated with three-dimension conformal radiotherapy,standard chemotherapy and endocrine therapy after operation.3 years after partial removal of breast,a retrospective historical control study had been fulfilled to analyze breast cosmetic results between two groups.Results In direct suture group,the cosmetology score was 4.3 points and fine rate was 40.9% (9/22).In breast reconstruction group.the score was 4.8 points and fine rate was 71.4% (15/21).The breast cosmetic effect in breast reconstruction group was significantly better than that in direct suture group (P<0.01).Conclusion It will get better breast cosmetic results after operation to utilize oncoplastic volume displacement suture technique in breast-conserving surgery.

C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

Objective To analyze the correlation among serum total IgE(TIgE),blood eosinophil(EOS) counts,the degree of sensitization for skin prick test(SPT),and fractional exhaled nitric oxide(FeNO)in chil-dren with asthma and allergic rhinitis. Methods We collected 121 children with asthma and allergic rhinitis andtheir serum TIgE,blood EOS,FeNO,and SPT for 14 common allergens. Results There were 75.2%patients sensitizing to at least two allergens.TIgE levels in poly-sensitized subjects were higher than that of mono-sensitized subjects(P < 0.05)The number of SPT sensitized allergens was positively correlated with the TIgE levels(P <0.05)FeNO levels were correlated with TIgE levels and EOS percentage(P < 0.05). TIgE levels in subjects with class 2 or 3 in SPT for house dust mite increased significantly ,compared with class 0 or 1. Conclusion There were some correlations among the four detections ,providing important reference for clinical diagnosis and treatment of respiratory allergic disease

Objective To explore the importance effect of circulating endothelial cells(CEC)in the pathogenesis of Henoch-Schonlein purpura(HSP)by determining the level of CEC in children with HSP,and to explore the relationship between CEC and therapeutic effect of HSP and the relationship between CEC and Henoch-Schonlein purpura nephritis(HSPN).Methods Sixty-six inpatients with HSP and 30 children as healthy control group were selected in the First Affiliated Hospital of Zhengzhou University from May to Dec.2008.The blood of all children,including the children in healthy control group,were sampled 1 mL in the early morning.And the CEC of all children were measured by flow cytometry method.The variation of CEC was analyzed in patients with HSP at acute stage and in healthy control group,in patients with HSP and in patients with HSPN,and in patients with HSP without renal damage at recovery stage and in patients with HSPN at recovery stage.SPSS 13.0 statistical software was used to analyze the data.Results CEC in patients with HSP at acute stage[(47.934 5?16.902 6)%]was significantly higher than that in healthy control group[(37.436 7?10.200 7)%](t=2.900 P0.05).CEC in patients with HSPN at recovery stage[(50.258 8?12.824 2)%]was significantly higher than that in patients with HSP without renal damage at recovery stage [(40.864 0?8.165 2)%](t=2.906 P

This research aimed at studying the effects of irrigation and rhizome length on the survival of ratio, yield and quality of Glycyrrhiza uralensis in wild tending condition. Employed the split-block design to carry out the field experiment, sampled with the quadrat method to measured the relative growth indexes and to estimate the yield, used the HPLC (high performance liquid chromatog- raphy ) method to measure the glycyrrhizin in the rhizome and adventitious root of the G. uralensis in this study. The quantity of the adventitious roots and the survival ratio were increased significantly as the length of the rhizome increased (P < 0.01), but the length of the rhizome had no remarkable effect on the content of glycyrrhizin. The average content of the glycyrrhizin in the adventitious root and rhizome could reach 3.03% and 2.12% after 3-year wild tending, respectively, and this results indicated that the quality of the glycyrrhiza using this method was much better than that from cultured glycyrrhiza with the reproducing method of seeding. so using the rhizome as reproductive material to produce the glycyrrhiza under the wild tending condition could get the high quality glycyrrhiza quick- ly and steadily, this phenomenon could be explained by the Hypothesis of synthetic inertia of the medicinal components from the wild material of G. uralensis. But the maximum yield with this method was just more than 945 kg x hm(-2) in this study. So the further work of how to increase the yield in the practical application with the method found in this study need to be done in the next research.
Agricultural Irrigation ; Culture Techniques ; methods ; Glycyrrhiza uralensis ; growth & development ; metabolism ; Glycyrrhizic Acid ; metabolism ; Rhizome ; growth & development ; Survival Analysis

OBJECTIVETo investigate the expression of ghrelin and its receptor, growth hormone secretagogue receptor (GHS-R), in the hypothalamus and gastrointestinal tract in rats with chronic renal failure (CRF) and explore their relationship with the disorder of gastrointestinal tract motility.

METHODSSD rats were randomly divided into sham-operated group (n=8) and CRF group (n=16), and in the latter group, the rats were subjected to 5/6 nephrectomy to induce CRF. Real-time PCR and immunohistochemical staining were used to detect the distribution of mRNA and protein of ghrelin and GHS-R in the gastric fundus, duodenum, and hypothalamus.

RESULTSThe rats in the CRF group showed a significantly higher expression of ghrelin mRNA and protein in the gastric fundus but a lower expression in the hypothalamus than those in the sham-operated group (P<0.01), but the expression in the duodenum was similar between the two groups (P>0.05). The expression of GHS-R mRNA and protein in the gastric fundus was significantly higher in the CRF group than in the sham-operated group (P<0.01), while in the hypothalamus and duodenum, the expression was significantly lower in the CRF group (P<0.01).

CONCLUSIONThe different distribution patterns of ghrelin and GHS-R in the tissues may be an important pathological basis of gastrointestinal motility disorder in CRF.

Animals ; Gastrointestinal Tract ; metabolism ; Ghrelin ; genetics ; metabolism ; Hypothalamus ; metabolism ; Kidney Failure, Chronic ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Ghrelin ; genetics ; metabolism

OBJECTIVETo investigate the underlying mechanism of the protective effects of resveratrol on oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes.

METHODSH9c2 cells, a permanent cell line derived from embryonic rat cardiac tissue, and then randomly divided into control group [PBS, cells exposed to H2O2 (600 µmol/L) for 20 min to induce mitochondrial oxidant damage], resveratrol group (0.01, 0.1, 1, 5, 10 and 20 µmol/L for 20 min at 20 min before exposing to H2O2), resveratrol plus inhibitor group (1 µmol/L KT5823 for 10 min at 10 min before 5 µmol/L resveratrol treatment) and inhibitor group (1 µmol/L KT5823 for 10 min). Mitochondrial membrane potential (ΔΨm) was measured by staining cells with tetramethylrhodamine ethyl ester (TMRE) and the mitochondrial permeability transition pore (mPTP) opening was evaluated by measuring the decrease of TMRE fluorescence intensity. Immunofluorescence assay was used to observe GSK-3β phosphorylation. The phosphorylation of GSK-3β and VASP were determined by Western blot. To detect intracellular NO, cells were loaded with DAF-FM DA (specific fluorescent dye of NO) and imaged with confocal microscopy.

RESULTSCompared to the control group, resveratrol (0.01-5 µmol/L) attenuated H2O2-induced mitochondrial damage reflected by attenuating the H2O2-induced TMRE fluorescence intensity decrease in a dose-dependent manner and the efficacy of 10 and 20 µmol/L resveratrol was significantly lower than that of 5 µmol/L resveratrol. Resveratrol also significantly upregulated the protein expression of VASP and increased GSK-3β Ser(9) phosphorylation, which could lead the inactivation of GSK-3β. These effects of resveratrol could be significantly abolished by protein kinase G inhibitor KT5823, while KT5823 alone did not affect GSK-3β and VASP phosphorylation. Confocal microscopy showed that DAF-FM (specific NO indicator) was similar between resveratrol and control group, suggesting that resveratrol did not produce NO.

CONCLUSIONSResveratrol could attenuate oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes by inactivating GSK-3β via cGMP/PKG signaling pathway independent of NO-related mechanism.

Animals ; Carbazoles ; pharmacology ; Cell Line ; Cyclic GMP ; metabolism ; Cyclic GMP-Dependent Protein Kinases ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Hydrogen Peroxide ; metabolism ; Mitochondria, Heart ; drug effects ; metabolism ; Myocytes, Cardiac ; cytology ; drug effects ; Oxidants ; metabolism ; Rats ; Signal Transduction ; drug effects ; Stilbenes ; pharmacology