Objective To investigate sexual function in male kidney transplant recipients and their offsprings health condition.Methods We studied 60 male kidney transplant recipients who had a normal sex life before kidney failure and had good renal graft function after transplant.The questionnaire was used to investigate the time of first spermatorrhea,viability of sex activities,frequency and sexual intercourse satisfaction of sex activities,procreation status,growth and intelligence status of the offspring.Semen routine tests before and after the transplantation were done.Results Hyposexuality or declined sexual function was found in all over 60 cases of recipients.And after surgery,sexual function of the recipients was improved obviously following the graft function gradually regained.Fifty-four patients had spermatorrhea in the first 1-3 months post-transplantation,and rest 6 cases had no spermatorrhea because of sexual life.After operation 56 male patients began to have sex life in 2-4 months with satisfaction rate of 86.7％ (52/60),except the other 4 cases who refused sex life due to worrying about graft damage by sex life.Sixty-eight children were born by 56 recipients after operation,and the growth and intelligence status had no significant difference from the children of the same age.No transplant graft failure or rejection was found after the recipients had children.Conclusion After kidney transplantation,the sexual function of male recipients could return to the normal status and the growth and intelligence status of the offspring is normal.

Objective To understand the feeling of family caregiver of children with chronic connective tissue disease (CTD).Methods A semi-structured interview was conducted with 30 family caregivers of children with CTD.Data were analyzed based on Colaizzi's phenomenological research method.Results The family caregiver's feelings were classified into four themes,including heavy psychological burden,diseases affecting family life,uncertainty in illness,demands for social support.Conclusions We should neglect the burden and related problems of caregivers when paying attentions to health problems of children,and supply effective intervention measures in order to increase the health level of children and their caregivers.

Objective To investigate the effects of transforming growth factorβ1(TGFβ1)and β3 (TGFβ3)gene transfer on MMP-2,MMP-9 and TIMP-1 expression in hepatic stellate cells(HSC-T6).Methods TGFβ1 and TGFβ3 expression plagmids were constructed.The recombinant expression plasmid pcDNA3.1(+)-=TGFβ1 and pcDNA3.1(+).TGFβ3 were transfected or cotransfected into HSC-T6.At 24,48 and 72 h after transfection,the expression of MMP-2,MMP-9 and TIMP-1 mRNA were detected by real-time quantitative PCR,and the expression of MMP-2,MMP-9 and TIMP-1 protein were detected by Western blot.The recombinant expression plasmid pcDNA3.1(+).TGFβ1 was transfected into HSC-T6,and positive clones were selected by G418.The positive clones were transfected by the recombinant expression plasmid pcDNA3.1(+).TGFβ1,and the expression of MMP-2,MMP-9 and TIMP-1 were detected at 48 h after transfection.Results After transfection with peDNA3.1-TGFβ1,MMP-2 and TIMP-1 increaged remarkably in HSC-T6 cells(P＜0.05),but MMP-9 remained at the sanle level;After transfection with pcDNA3.1-TGFβ3,expression levels of MMP-2,MMP-9 and TIMP-1 mRNA were not changed,but TIMP-1 protein increased remarkably(P＜0.05);in cotransfection group,the expression of MMP-2 was higher than that in the blank and the control groups(P＜0.05),but MMP-9 level was not changed and TIMP-1was decreased compared with that in the TGF-β1 transfection group(P＜0.05).After TGFβ3was transfected into positive clones,the change of MMP-2 wag not significant(P＞0.05).but MMP-9 increaged and TIMP-1 decreased significantly at 48 h after transfection(P＜0.05).Conclusions TGFB3 may inhibit liver fibrosis by increase the activity of MMP-2 and MMP-9,and decrease the activity of TIMP-1.

Objective To establish Beagle dogs’model of abducens nerves injury and to observe the clinical therapeu?tic effect of electroacupuncture treatment. Methods Twenty-four Beagle dogs were randomly divided into simple crush group (control group) and crush with electrical stimulation group (experimental group). Cisternal segment of the abducens nerve was given a crush injury, then electrodes were implanted to stimulate the abducens nerve and lateral rectus muscle. Distance between the center of the pupil to medial margin of extraocular adjoin was measured from 1 to 12 weeks after opera?tions. Results All procedures used in the study were well tolerated by Beagle dogs. Electrode implantation to stimulate the lateral rectus muscle and the abducens nerve behind of cavemous sinus was successful. There was no statistical significance of the distance between the two groups from 1 to 2 weeks after operations, and the distance was shorter in experimental group than that in control group from 4 to 12 weeks after operations (P<0.01). Conclusion The animal models established to study electroacupuncture treatment of the injured abducens nerves was successful. Electroacupuncture can promote the re?covery of the injured abducens nerves obviously.

Objective To investigate the effect of jejunal infusion of amino acids on secretion of gastrointestinal hormone in healthy dogs.Methods Six healthy adult dogs were treated with jejunal fistulas and femoral vein intubation.Twenty-four hours after the operation,solution of 8 different amino acid monomers (experimental group) or normal saline (control group) were infused into the jejunum of the dogs every 24hours.The levels of cholecystokinin (CCK),motilin,and gastrin in the peripheral plasma were measured using radioimmunoassay at the start of infusion (0 minute),and 30,60,90,and 120 minutes after infusion.Results Compared with the control group,the serum CCK level in the phenylalanine group was significantly higher 30 and 60 minutes after infusion [(1.25 ±0.19) ng/L vs.(0.66 ±0.14) ng/L,(1.23 ±0.12) ng/L vs.(0.80 ± 0.03) ng/L,both P ＜ 0.01],while that in the tryptophan group was significantly higher 30 minutes after infusion [(1.08 ±0.26) ng/L vs.(0.66 ±0.14) rig/L,P ＜0.01].The other measurement results showed no statistically significant differences.Conclusions Jejunal infusion of phenylalanine or tryptophan may stimulate the secretion of gastrointestinal hormone to some extent.Aromatic amino acids (phenylalanine and tryptophan) is more potent in triggering the release of CCK than aliphatic (leucine,isoleucine,and methionine) and charged amino acids (aspartic acid,arginine,and glutamate).The mechanism may be related to the properties of the amino acids.

Objective To evaluate the radiosensitization and mechanism of DNA-dependent protein kinase catalytic subunit inhibitor NU7026 on HCT116 colorectal cancer stem cells.Methods The flow cytometry was used to determine the sub-population of cancer stem cells with the markers CD133/ CD44.The cells were divided into four groups:control group,20 μmol/L NU7926 group,2 Gy irradiation group,and 20 μmol/L NU7026 combined with 2 Gy irradiation group.Cell proliferation and survival were evaluated by colony-formation experiment.Flow cytometry was used to analyze the cell cycle distribution and apoptosis induction.γ-H2AX foci were detected with immune-fluorescence staining analysis by a laser confocal microscopy for investiating the DNA double-strand break repair.Results The flow cytometric data of CD133 +/CD44 + positive cells indicated that the sub-population of cancer stem cell (CSC) took the ratio of (88.14 ± 0.47)％ of the cultured HCT116 cells.The colony-formation efficiency of HCT116 cells was (84.75 ± 1.35) ％ in serum-free mediums in vitro culture.Compared to 2 Gy irradiation alone group,the NU7026 combined with 2 Gy irradiation group had a lower cell colony-forming ratio (t =7.21,P ＜0.01) and a lower survival ratio (t =7.22,P ＜ 0.01).The proportion of CSCs sub-population increased at 48 h post-2 Gy irradiation,suggesting that HCT116 CSCs was more resistant to ionizing radiation.Importantly,NU7026 largely decreased the proportion of CSCs sub-population in 2 Gy-irradiated cells.The difference of CSC proportion between 2 Gy irradiation alone group and 2 Gy combined with NU7026 treatment group was statistically significant (t =9.55,P ＜ 0.01).In addition,the group of NU7026 combined with 2 Gy irradiation had a higher ratio of G2/M arrest 24 h post-irradiation (t =7.67,P ＜ 0.01) and an increased induction of cell early apoptosis (t =8.24,P ＜ 0.05).48 h post irradiation as compared to 2 Gy irradiation alone group.NU7026 treatment significantly inhibited the cellular capacity of repairing DNA double-strand breaks induced by γ-ray irradiation.The γ-H2AX foci of the combined treatments group were much higher than that of 2 Gy irradiation alone group at 2,4,8,24 h postirradiation (t=19.58,11.95,7.01,9.45,P＜0.01).Conclusions DNA-dependent protein kinase catalytic subunit inhibitor NU7026 can significantly sensitize the cancer stem cells of colorectal carcinoma HCT116 cells to γ-ray irradiation.Multiple mechanisms are involved in the radiosensitization effect of NU7026,including DNA repair inhibition,elongation of G2/M arrest,and increase of radiation-induced apoptosis.

Objective To evaluate the relationship between the mechanism underlying docosahexaenoic acid (DHA)-induced regulation of angiopoietin expression and Src-suppressed C kinase substrate (SSeCKS) in human brain vascular pericytes (HBVPs) subjected to oxygen-glucose deprivation and restoration (OGD/R).Methods HBVPs were seeded in 96-well or in 6-well plates at a density of 2× 105 cells/ml and divided into 5 groups (n =18 each) using a random number table:control group (group C),OGD/R group,DHA group (group D),SSeCKS gene silencing group (group S) and SSeCKS gene silencing plus DHA group (group SD).The model of OGD/R injury was established as follows:the cells were subjected to O2-glucose deprivation for 24 h in glucose-and serum-free culture medium aerated with 94％ N2-5％ CO2-1％ O2 followed by restoration of O2-glucose supply for 6 h in high-glucose DMEM culture medium in normal atmosphere.DHA was added at 1 h before hypoxia with the final concentration of 40 μmol/L in group D.Small interfering RNA induced SSeCKS gene silencing in S and SD groups.Subsequently,DHA with the final concentration of 40 μmol/L was added at 1 h before hypoxia in group SD.At 6 h of reoxygenation,the cell survival rate was determined by CCK-8 assay,the amount of LDH released was detected using ELISA,and the expression of SSeCKS,angiopoietin-1 (Ang-1) and Ang-2 was detected by Western blot.Results Compared with group C,the cell survival rate was significantly decreased,the amount of LDH released was increased,the expression of SSeCKS and Ang-1 was down-regulated,the expression of Ang-2 was up-regulated,and Ang-1/Ang-2 ratio was decreased in group OGD/R,and the expression of SSeCKS was down-regulated in group S (P＜0.05).Compared with group OGD/R,the cell survival rate was significantly increased,the amount of LDH released was decreased,the expression of SSeCKS and Ang-1 was up-regulated,the expression of Ang-2 was down-regulated,and Ang-1/Ang-2 ratio was increased in group D (P＜O.05),and no significant change was found in the parameters mentioned above in group SD (P＞0.05).Compared with group D,the cell survival rate was significantly decreased,the amount of LDH released was increased,the expression of SSeCKS and Ang-1 was down-regulated,the expression of Ang-2 was up-regulated,and Ang-1/Ang-2 ratio was decreased in group SD (P＜0.05).Conclusion The mechanism by which DHA increases the ratio of Ang-1/Ang-2 may be totally related to up-regulation of SSeCKS expression in HBVPs subjected to OGD/R.

Objective To study the physical and chemical properties of silibinin; To lay foundation for optimal formulation design.Methods The high performance liquid chromatography method was used to detect the equilibrium solubility of silibininin in various solutions. The partition coefficients of silibinnin in the n-octalution-water and n-octalution-buffer solution systems were determined by shaking flask method. The destructive tests were carried out on silibinin.Results The equilibrium solubility of silibininin at 25℃ was 1.352 mg/L in water and the largest in acetonitrile, and increased in basic buffer solutions and after adding surfactants, of which sodium dodecyl benzene sulfonate was the strongest in solubilizing ability. The apparent oil-water partition coefficient of silibinnin was 61.39. Silibinnin was unstable in the acidic, basic, oxidizing and reducing solutions.Conclusion The experiment results can provide references for designing new preparations of silibinin.

Objective To explore the features of the cytochrome c oxidase subunit Ⅰ (COXⅠ) of Asian black bear Sichuan subspecies. Methods Primers were designed according published gene sequences of other bear species. A DNA sequence was amplified by PCR from the bear Sichuan subspecies’s hair-follicle tissue total DNA, then the PCR product was sequenced. The results were analyzed and a forecast analysis was carried on. Results The amplified COXⅠ gene of Sichuan black bear was 1 605 bp and contained a 1545-nucleotide open reading frame encoding 514 amino acid residues. The overall base composition of COXⅠ gene is 26.9 %, 23.1%, 18.3 %, and 31.8 % respectively to A, C, G, and T. The start codon of the COXⅠ is ATG, while the termination codon TAA. The protein’s pI is 6.29 and weight is 57.2?10 3 . The similarity analysis and phylogenetic tree analysis indicated that Sichuan black bear has closer relationship to American black bear, but not to brown bear and polar bear. Among dogs, caws and rats, it is closer to the former one. Conclusion There are high similarities in the DNA sequence, protein amino acid sequence of Asian black bear Sichuan subspecies with other bear species, indicating a high conservativity of COXⅠ in these involved species.

Objective To investigate the sequence characteristic of black bear's mitochondrial NADH-ubiquinone oxidoreductase chain 4 gene(ND4).Methods Primers were designed according to certain species' NADH-ubiquinone oxidoreductase ND4 in which the DNA sequence had been reported.The DNA sequence was amplified by PCR from the Asian black bear Sichuan subspecies's(Ursus thibetanus mupinensis)hair-follicle tissue total DNA and were analyzed with its amino acid sequence.Results The ND4 gene sequence length was 1 402 bp and contained a 1 377-nucleotide open reading frame encoding 459 amino acid residues.The amino acid sequence's pI was 9.37 and weight was 51.5?10 3.The DNA sequence and corresponding amino acid sequence of black bear's ND4 gene had the high similarity with those of other mammalian animals' ND4 gene,and the similarity was 89% to 91% and 94% to 95%.The result of phylogenetic tree was basically consistent with the traditional species system phylogenetic relations.Topology prediction showed the Asian black bear Sichuan subspecies's ND4 proteins contained N-glycosylation site,kinase C phosphorylation sites,casein kinase Ⅱ phosphorylation site,N-myristoylation site and leucine-rich region.In addition,black bear's ND4 protein contained leucine zipper pattern and Leucine-rich region.Conclusion The mammalia animal's ND4 gene has the high conservative nature.The pretein coded by ND4 gene has high uniformity in the function.