OBJECTIVETo investigate the role of advanced glycation end products (AGEs) and their receptors (RAGE) in the pathogenesis of diabetic mellitus erectile dysfunction (DMED) and the effects of AGEs and RAGE on the activity of endothelin-1 (ET-1) in rat cavernosum.

METHODSForty male Sprague-Dawley rats were taken at random to construct 2 groups of diabetes mellitus (DM) models of equal number, one given free access to water and the other administered aminoguanidine hydrochloride (DM + AG) in water at the dose of 1 g/L. Another 20 male SD rats were equally divided into a normal control and an AG control group. After 8 weeks, the cavernosum tissues were harvested from all groups of rats, part of the isolated penile tissues homogenated to detect the content of AGE-peptide (AGE-P) and the activity of ET-1, and the AGEs and RAGE in the rest of the penile tissues analyzed by immunohisto- chemical assay.

RESULTSCompared with the normal controls, the expressions of AGEs and RAGE, the content of AGE-P and the activity of ET-1 in the cavernosum tissues were significantly high in the DM group (P < 0.05), while the administration of AG to the DM rats reversed the above results. No significant difference was observed between the normal control and AG control groups in any of the data (P > 0.05).

CONCLUSIONIn DM conditions, the joint effect of AGEs and RAGE may elevate the activity of ET-1 in rat cavernosum and thus promote the development of DMED.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; prevention & control ; Endothelin-1 ; metabolism ; Enzyme Inhibitors ; administration & dosage ; Glycation End Products, Advanced ; antagonists & inhibitors ; metabolism ; Guanidines ; administration & dosage ; Male ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; metabolism

OBJECTIVETo explore the methodology as well as the features, quantificational index, and reference standard of 3.0 Tesla magnetic resonance (MR) diffusion-weighted imaging (DWI) on the normal rabbit's liver.

METHODSTwenty New Zealand white rabbits were enrolled and DWI was performed after anesthetics with multi-b values at 3.0 T MR scanner. Apparent diffusion coefficient (ADC) values as well as the difference between maximum and minimum ADC values, signal strength (SH), noise signal (SD), signal to noise ratio (SNR), and quality index (QI) were recorded and analyzed.

RESULTSWith b value increased, the ADC values decreased accordingly (P < 0.001). The difference between maximum and minimum ADC values with b = 1000 s/mm2 was the least (good stability), b = 600 s/mm2 was the second least, and b = 300 s/mm2 was greatest (bad stability). The SH decreased at the same time (P < 0.001), but the difference among DWI with b =600, 800, and 1000 s/mm2 was not statistically significant (P > 0.05). The SD decreased at the same time (P < 0.001), but the difference between DWI with b = 800 s/mm2 and b = 1000 s/mm2 was not statistically significant (P > 0.05). The SNR decreased at the same time (P < 0.001), but there were no significant differences between DWI with b = 600 s/mm2 and b = 800 s/mm2 or b = 600 s/mm2 and b = 1000 s/mm2 (P > 0.05). The SNR of DWI with b = 800 s/mm2 and b = 1000 s/mm2 was lower. The QI decreased at the same time (P < 0. 001) , but the difference between DWI with b = 800 s/mm2 and b = 1000 s/mm was not statistically significant (P > 0.05).

CONCLUSIONWhen 3.0 T MR DWI is applied for rabbit liver, it is better to use b = 600 s/mm2 for reducing scanning time and assuring better diffusion weights, quantity of images, and stability of ADC measurement.

Animals ; Diffusion Magnetic Resonance Imaging ; methods ; Liver ; Male ; Rabbits

OBJECTIVETo observe the protective effect of Shenfu Injection (SFI) pretreatment on brain of patients receiving aortic valve replacement (AVR) undergoing cardiopulmonary bypass (CPB).

METHODSThirty AVR patients undergoing CPB were randomly assigned to 2 groups, the control group and the experimental group, 15 cases in each group. SFI at 1.5 mL/kg (dissolved in 250 mL 5% glucose solution) was intravenously dripped to those in the experimental group 5 days before operation, once daily for 5 successive days. SFI at 1.5 mL/kg (dissolved in 250 mL 5% glucose solution) was intravenously dripped to those 30 min before anesthesia induction. Equal dose of normal saline was intravenously dripped to those in the control group, and the other procedures were the same as those for patients in the experimental group. The venous blood sample (2 mL) was drawn from the right internal carotid vein immediately after induction of anesthesia (T1),10 min after CPB (T2), 30 min after GPB (T3), 2 h after CPB (T4), 24 h after CPB (T5), and 48 h after CPB (T6), thus detecting the plasma levels of S100beta and neuron specific enolase (NSE). And patients' cognitive function was assessed with mini-mental state examination (MMSE) scale on the day before operation, the 2nd and the 7th day after operation.

RESULTSThere was no statistical difference in the levels of S1001 and NSE between the two groups at T1 (P > 0.05). There was statistical difference in the levels of S100beta and NSE between the two groups at T2, T3, T4, T5, T6, when compared with those at T1 (P <0.05). Besides, the levels of S100beta and NSE at T2, T3, T4, T5, T6 were lower in the experimental group than in the control group, showing statistical difference (P <0.05). The MMSE scores decreased on the 2nd day after operation in the two groups, showing statistical difference when compared with those on the day before operation (P <0.05). It was lowered more obviously in the control group. There was no statistical difference in the MMSE score between the 7th day post-operation and the day before operation (P >0.05).

CONCLUSIONSFI pretreatment had protective effect on brain in AVR patients undergoing CPB.

Brain ; drug effects ; metabolism ; Cardiopulmonary Bypass ; Cognition ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Intraoperative Period ; Ischemic Preconditioning ; methods ; Male ; Middle Aged ; Phosphopyruvate Hydratase ; metabolism ; S100 Calcium Binding Protein beta Subunit ; metabolism

Objective To explore the effects of stress and self-efficacy on the quality of life in patients with gallstones and the effect relationship of self-efficacy in the quality of life and stress, and provide guidance for the health intervention of patients with gallstones. Methods Totally 362 patients with gallstones after operative that already returned to the community for 12 months were investigated, and their stress condition, self-efficacy and quality status of life were measured by the life events scale ( LES ) , self-efficacy scale and gastrointestinal quality of life index (GIQLI). The effect relationship of self-efficacy in the quality of life and stress was analyzed.Results The self-efficacy played partial mediating effect between stress and quality of life. When the intermediary variables-self-efficacy sense was participated in, the regression coefficient between stress and quality of life was significantly reduced (β was reduced from -0. 695 to -0. 548 ) . Self efficacy played regulating effect between stress and quality of life, which means that self efficacy sense × stress partial regression coefficient in the regression which regard the quality of life as the dependent variable can reach to a significant level (β=-0.086, P<0.05) , and introduced interaction terms after new interpretation (△R2 ) also can reach to a significant level (△R2 = 0. 017, P<0. 05 ) . Conclusions Self efficacy played the mediating effect and regulating effect in the relationship between stress and quality of life. Nursing staff can reduce the negative effects of stress events in patients′quality of life through the interventions in patients′self-efficacy.

Objective To inverstigate the effects of follow-up management based on social goals model on social relation quality in hepatocellular carcinoma patients after transcatheter arterial chemoebolization (TACE).Methods By convenience sampling method, a total of 113 patients after TACE were divided into the control group ( n=56) and the intervention group ( n=57) according to the treatment time. In the control group, routine follow-up based on case-management was adopted, while in the intervention group, the follow-up based on social goals model was adopted. The intervention effect in two groups were tested by social relation quality scale.Results After intervention, the total score of social relation quality, score of family adaptability and score of family commitment were (53.60±8.86),(20.34±3.28) and (18.07±2.96), and they were all significantly higher than the scores in the control group [(49.25±9.75),(18.52±3.55),(16.22±3.21)](P<0.05). Conclusions The follow-up management based on social goals model can effectively improve the level of family adaptability and family commitment. The intervention could be one of the effective ways to implement social relational quality fin patients after TACE.

OBJECTIVETo investigate the binding mechanisms of FVIII Trp1707Ser mutation-associated inhibitor.

METHODSThe APPT, PT, TT, Fg and FVIII:C were detected to make phenotypic diagnosis of haemophilia A. Inhibitors titer were measured by Bethesda method. Long distance-PCR (LD-PCR) and sequence-specific PCR were adopted for screening the intron 22 and intron 1 inversions respectively. FVIII coding and boundary sequences were analyzed by direct DNA sequencing. Inhibitor was reacted with different segments of FVIII, including heavy chain and its components A1 and A2, light chain and its components A3, C1 and C2. Corrected test was used to measure the remaining F VIII:C (% ) by adding pooled normal plasmas. After labeling purified inhibitors with biotin, western blot was performed to further confirm the binding reactions between inhibitors and segments.

RESULTSThe haemophilia A patient had mild deficiency of FVIII:C (1.1%) and had high FVIII inhibitor titer of 18.4 BU. A mutation c.97223C>G in exon 14 of F8 gene resulted to p.Trp1707Ser was identified by DNA sequencing. Corrected test showed that the remaining F VIII:C was increased when inhibitors reacted with heavy chain and light chain, especially with heavy chain. The remaining FVIII:C was also increased in the A2 and C2 domain reactions. No significant differences were seen in the A1, A3 and C1 domain reactions. Antigen-antibody reaction bands were confirmed by western blots when degenerated B-domain deleted recombinant FVIII, A2 and C2 were used as antigens.

CONCLUSIONThe binding sites of FVIIITrp1707Ser mutation inhibitor were the A2 domain of heavy chain and C2 domain of light chain. The binding reaction with heavy chain was more intense.

Binding Sites ; genetics ; Exons ; Factor VIII ; antagonists & inhibitors ; genetics ; Hemophilia A ; genetics ; Humans ; Male ; Mutation ; Young Adult

BACKGROUNDThe primary reasons for local recurrence and therapeutic failure in the treatment of malignant gliomas are the invasion and interactions of tumor cells with surrounding normal brain cells. However, these tumor cells are hard to be visualized directly in histopathological preparations, or in experimental glioma models. Therefore, we developed an experimental human dual-color in vivo glioma model, which made tracking solitary invasive glioma cells possible, for the purpose of visualizing the interactions between red fluorescence labeled human glioma cells and host brain cells. This may offer references for further studying the roles of tumor microenvironment during glioma tissue remodeling.

METHODSTransgenic female C57BL/6 mice expressing enhanced green fluorescent protein (EGFP) were crossed with male Balb/c nude mice. Then sib mating was allowed to occur continuously in order to establish an inbred nude mice strain with 50% of their offspring that are EGFP positive. Human glioma cell lines U87-MG and SU3 were transfected with red fluorescent protein (RFP) gene, and a rat C6 glioma cell line was stained directly with CM-DiI, to establish three glioma cell lines emitting red fluorescence (SU3-RFP, U87-RFP, and C6-CM-DiI). Red fluorescence tumor cells were inoculated via intra-cerebral injection into caudate nucleus of the EGFP nude mice. Tumor-bearing mice were sacrificed when their clinical symptoms appeared, and the whole brain was harvested and snap frozen for further analysis. Confocal laser scanning microscopy was performed to monitor the mutual interactions between tumor cells and host brain cells.

RESULTSAlmost all the essential tissues of the established EGFP athymic Balb/c nude mice, except hair and erythrocytes, fluoresced green under excitation using a blue light-emitting flashlight with a central peak of 470 nm, approximately 50% of the offsprings were nu/nu EGFP+. SU3-RFP, U87-RFP, and C6-CM-DiI almost 100% expressed red fluorescence under the fluorescence microscope. Under fluorescence microscopic view, RFP+ cells were observed growing wherever they arrived at, locating in the brain parenchyma, ventricles, and para-vascular region. The interactions between the transplanted tumor cells and host adjacent cells could be classified into three types: (1) interweaving; (2) mergence; and (3) fusion. Interweaving was observed in the early stage of tumor remodeling, in which both transplantable tumor cells and host cells were observed scattered in the tumor invading and spreading area without organic connections. Mergence was defined as mutual interactions between tumor cells and host stroma during tumorigenesis. Direct cell fusion between transplantable tumor cells and host cells could be observed occasionally.

CONCLUSIONSThis study showed that self-established EGFP athymic nude mice offered the possibility of visualizing tumorigenesis of human xenograft tumor, and the dual-color xenograft glioma model was of considerable utility in studying the process of tumor remodeling. Based on this platform, mutual interactions between glioma cells and host tissues could be observed directly to further elucidate the development of tumor microenvironment.

Animals ; Cell Line, Tumor ; Female ; Glioma ; metabolism ; pathology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Nude ; Mice, Transgenic

OBJECTIVETo analyze the phenotype and genotype of two Chinese pedigrees with von Willebrand diseases, and to investigate the molecular pathogenesis.

METHODSBleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor-ristocetin cofactor (vWF:Rco), von Willebrand factor antigen (vWF:Ag), von Willebrand factor activity (vWF:A), von Willebrand factor collagen binding assay (vWF:CB) and multimer analysis were used for phenotype diagnosis. DNA was extracted. All of the 52 exons and exon-intron bounda ries of the VWF gene were amplified with polymerase chain reaction(PCR) and analyzed by direct sequencing.

RESULTSAPTT and BT were prolonged. Plasma RIPA, vWF:Rco, vWF:Ag, vWF:A and vWF:CB was significantly decreased. No VWF multimer can be found by plasma VWF multimer analysis. Homozygous insertional mutation g.82888_82889insCATG in exon 17 was found in proband A. Compound heterozygous mutations g.94865 G to A (Trp856stop) in exon 20 and g.110698_110699delinsG in exon 28 were found in proband B.

CONCLUSIONHomozygous insertional mutation g.82888_82889insCATG and compound heterozygous mutations g.94865G to A(Trp856X) and g.110698_110699delinsG probably have respectively induced type 3 von Willebrand diseases in the two probands.

Adolescent ; Female ; Genotype ; Humans ; Male ; Mutation ; Pedigree ; Phenotype ; von Willebrand Disease, Type 3 ; genetics ; von Willebrand Factor ; genetics

OBJECTIVETo investigate the function of abnormal fibrinogen in two inherited dysfibrinogenemia pedigrees.

METHODSRoutine coagulation tests were conducted in the probands and related family members. The antigen and activity levels of fibrinogen were detected by immunoturbidimetry assay and clauss assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes and antithrombin gene（AT3）were analyzed by PCR amplification and direct sequencing. Routine thrombelastography (TEG) test and functional fibrinogen TEG test were both used to make a comprehensive evaluation of coagulation status and functional fibrinogen level in patients. The molecular weights of the three peptides from fibrinogen were measured by Western blot. The function of abnormal fibrinogen was assessed by fibrinogen dynamic polymerization and fibrinolysis velocity.

RESULTSThe coagulation routine tests were normal in two probands except for prolonged thrombin time (TT) and reptilase time (RT), as well as reduced activity levels of 0.5 g/L and 0.6 g/L fibrinogen, respectively. The antigen levels of fibrinogen were 2.32 g/L and 2.66 g/L in two probands, which were in the normal reference range. The genotype analysis showed that Arg275His in fibrinogen γ chain (γ Arg275His) existed in both probands and patients in these two pedigrees. Meanwhile, proband B's grandfather and aunt also carried heterozygote g.5876T>C (Ser116Pro) mutation in AT3. The results of routine TEG test demonstrated that the α values of proband B and his father were close to and lower than the lower limit of reference range, respectively, while the MA values were normal in both of them. However, functional fibrinogen TEG test revealed obviously reduced MA value. All the probands and patients demonstrated prolonged lag-off time and reduced peak value in fibrinogen dynamic polymerization tests. Meanwhile, most of fibrin formed from the patients' plasma could not be dissolved completely by plasminogen (PLG) and urokinase-typeplasminogenactivator (u-PA) at a certain time.

CONCLUSIONWe first reported cases of inherited dysgibrinogenemia associated with inherited AT deficiency. γArg275His mutation caused the abnormal fibrinogen in terms of fibrin mono polymerization and possibly in fibrinolysis. Combined use of routine TEG test and functional fibrinogen TEG test with comprehensive analyses of the parameters in both tests could better evaluate the level of functional fibrinogen and predict the risk of hemorrhage and thrombosis in patients with inherited dysfibrinogenemia.

Adult ; Afibrinogenemia ; genetics ; Child ; Female ; Fibrinogen ; genetics ; Fibrinogens, Abnormal ; genetics ; physiology ; Genotype ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

OBJECTIVETo explore the mutations of coagulation factor VII (FVII) gene in one pedigree with hereditary FVII deficiency, and to investigate the molecular mechanisms of FVII deficiency.

METHODSFVII gene mutations were analysed in the pedigree by direct DNA sequencing. The mutated DNA fragments were cloned into pMD19-T simple TA vector, and sequenced to confirm their distribution on chromosome. The plasma activity of FVII of the probands and their family members was detected with coagulation assay. The antigen of FVII were identified with ELISA.

RESULTSThree gene mutations were detected in the pedigree: A/G to C at 15386 resulting in Arg353Pro/Gln353Pro, A to T at 15274 resulting in Lys316Stop, all three mutations were heterozygotes. Three kinds of polymorphisms were identified in his father: A to G transition at position 15386 resulting in Arg353Gln, heterozygotic deletion of 2050 - 2059 cctatatcct in promoter and G to A mutation in intron 1a, the same polymorphisms were found in his grandfather. The three polymorphisms were located in the same chromosome of his father.

CONCLUSIONTwo mutations were found in the pedigree with hereditary FVII deficiency. One is nonsense mutation (Lys316Stop), the other is missense one (Gln353Pro). Gln353Pro and Lys316Stop might be the molecular mechanisms of FVII deficiency. The two novel mutations were reported for the first time in the literature.

Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Heterozygote ; Humans ; Male ; Mutation, Missense ; Pedigree ; Polymorphism, Genetic ; Sequence Deletion