OBJECTIVETo investigate the role of endogenous hydrogen sulfide (H2S) in erectile dysfunction (ED) induced by androgen deficiency.

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into six groups: 2-week control (A), 4-week control (B), 2-week castration (C), 4-week castration (D), 2-week castration + androgen replacement (E), and 4-week castration + androgen replacement (F), those in groups E and F subcutaneously injected with testosterone propionate (TP) at the physiological dose of 3 mg/kg per day after castration, while those in the other groups with isodose oil instead. At 2 and 4 weeks after operation, we determined the level of serum testosterone (T) , intracavernous pressure (ICP) , mean carotid arterial pressure (MAP) of the rats, measured the concentration of H2S in the plasma and corpus cavernosum tissue, and detected the expressions of cystathionine-P3-synthase (CBS) and cystathionine-gamma-lyase (CSE) by immunohistochemistry and Western blot.

RESULTSThe serum T level was significantly lower in group C ([0.63 +/- 0.15] nmol/L) than in A ( [ 16.55 +/- 4.17] nmol/L) and E ( [ 18.99 +/- 4.62] nmol/L) (P <0.05), as well as in group D ([0.70 +/-0.22] nmol/L) than in B ([15.44 +/-5.18] nmol/L) and F ([20.99 +/-6.41] nmol/L) (P <0. 05) , and so were ICP/MAP after 5 and 7 V electrical stimulation of the pelvic ganglia (P <0. 05) , H2 S concentration (P <0.05), and the expressions of CBS and CSE (P <0.05). The expressions of CBS and CSE proteins were also significantly decreased in group C as compared with D (P <0.05).

CONCLUSIONThe reduced expressions of CBS and CSE may inhibit the H2 S signaling pathway, which might be one of the mechanisms underlying androgen deficiency-induced ED in rats.

Androgens ; deficiency ; Animals ; Cystathionine beta-Synthase ; metabolism ; Cystathionine gamma-Lyase ; metabolism ; Erectile Dysfunction ; metabolism ; Hydrogen Sulfide ; metabolism ; Male ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study incidence and death among previous paid blood-donated AIDS sufferers.

METHODSA retrospective cohort study was adopted to study incidence and death of 373 previous paid blood-donated HIV sufferers and its effect factors.

RESULTSPrevious paid blood-donated HIV infection was serious and the infection rate in blood-donated crowd was 35.87% (373/1040); the mean incubation period of AIDS was 8.87 years (95% CI: 8.76 - 8.99, Kaplan-Meier method); the cumulative incidence of AIDS (10 years) was 92.23% (344/373), and the incidence of total sufferers was 11.64/100 person-year; the cumulative probability of survival of one-year, three-year, five-year AIDS sufferers was separately 94.48% (325/344), 85.76% (295/344) and 83.14% (286/344), median survival time was over 5 years; the anti-virotic treatment days (960.29 +/- 486.38), infection age (33.39 +/- 9.08) disease age (41.98 +/- 8.88) had significant effects on AIDS sufferers' survival time/survival rate (chi(2) = 61.355, P = 0.000; chi(2) = 6.555, P = 0.010; chi(2) = 3.969, P = 0.046).

CONCLUSIONThe survival time of previous paid blood-donated HIV cases was longer, and their survival rate was higher, remarkably higher than the UNAIDS' research findings.

Acquired Immunodeficiency Syndrome ; epidemiology ; mortality ; Adult ; Age of Onset ; Blood Donors ; China ; epidemiology ; Cohort Studies ; Female ; Follow-Up Studies ; Humans ; Incidence ; Male ; Middle Aged ; Retrospective Studies ; Rural Population ; Surveys and Questionnaires ; Survival Rate

OBJECTIVETo construct the yeast two-hybrid system, and screen the proteins which interact with FasL, and investigate the relationship of FasL and hepatic metastasis of colorectal carcinoma.

METHODSWe have cloned the FasL gene into the pGBKT7 vector as the bait, then screened the fetal liver cDNA library, and have got a series of specific proteins that interact with FasL protein. Using the bioinformatics, we analyzed the interacting proteins in the mechanism of hepatic metastasis of colorectal carcinoma.

RESULTSWe have screened several proteins that interaction with FasL protein, including metallothionein 1K, 1G, 2A, cathepsin B, fatty acid synthase, interferon alpha-inducible protein 27, phospholipid scramblase, Ser/Thr-like kinase, anchor attachment protein, fibulin-5.

CONCLUSIONSWe have successfully constructed the yeast two-hybrid system, and preliminary identified that the interaction between FasL, metallothionein, cathepsin and anchor attachment protein is radically related to the hepatic metastasis of colorectal carcinoma.

Cathepsin B ; metabolism ; Cloning, Molecular ; Colorectal Neoplasms ; chemistry ; pathology ; Fas Ligand Protein ; Gene Library ; Humans ; In Vitro Techniques ; Liver Neoplasms ; chemistry ; secondary ; Membrane Glycoproteins ; genetics ; metabolism ; Metallothionein ; metabolism ; Protein Binding ; Tumor Necrosis Factors ; genetics ; metabolism ; Two-Hybrid System Techniques ; Yeasts ; genetics

OBJECTIVETo study the expression of metallothionein (MT) and FasL in colorectal cancer and their relation to lymph node and liver metastasis.

METHODSImmunohistochemistry and quantitative RT-PCR were used to detect expression of MT and FasL in protein and mRNA levels in 93 cases of colorectal cancer.

RESULTSThe rates of MT expression in primary foci, non-cancerous colon mucosa, lymph node metastasis and liver metastasis were 58.1%, 32.3%, 81.1%, 64.3% respectively. And the rates of FasL expression were 41.9%, 19.4%, 62.3%, and 92.9% respectively. The positive rates of MT and FasL in primary foci, liver and lymph node metastasis were higher than that in non-cancerous mucosa (chi(2) = 35.2421, 57.5152, P < 0.01). MT expression rate in lymph node metastasis was higher than that in primary foci (chi(2) = 8.0565, P < 0.01). In liver metastasis, FasL expression rate was higher than in lymph node metastasis and primary foci (chi(2) = 8.6674, 22.4455, P < 0.01). The positive rates of MT and FasL in Dukes stage C and D were higher than that in Dukes stage A and B (chi(2) = 18.8871, 25.1650, P < 0.01). And higher rates of MT and FasL expression were observed in low differentiation adenocarcinoma and mucus adenocarcinoma than in middle-high differentiation adenocarcinoma (chi(2) = 11.1546, 9.2239, P < 0.05). High MT mRNA level was found in lymph node metastasis and high FasL mRNA level in liver metastasis.

CONCLUSIONSEnhanced expression of MT and FasL was associated significantly with lymph node and liver metastasis of colorectal cancer. Assay of MT and FasL expression has prognostic values for colorectal cancer patients.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; metabolism ; pathology ; Fas Ligand Protein ; biosynthesis ; genetics ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; secondary ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Metallothionein ; biosynthesis ; genetics ; Middle Aged ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To establish an automatic synthesis method for 11C-carfentanil (CFN) as an novel opiate receptor radioligand and study its biodistribution in rats. Methods 11C-Triflate-CH3 was bubbled into 0.5 mg precursor desmethyl-CFN (which was dissolved in 0.15 ml DMSO) to generate 11C-CFN in a V-tube at room temperature. Sep-Pak C2 column was used for purification of 11C-CFN, which was eluted by 3ml binary system aqueous solution, 10 ml water thrice, and then I ml ethanol. The biodistribution (% ID/g) of 11C-CFN in SD rats was studied. SPSS 13.0 was used for statistical analysis. Non-normal distribution data were analyzed using nonparametric test. Results The synthesis time for 11C-CFN was 20 min (end of bombardment, EOB). The synthesis yield was (35.5 ± 2.2) % on average (n = 12, uncorrected)with the radiochemical purity over 98%. Biodistribution study in rats showed that the tracer had a high brain uptake, rapid blood clearance, and a metabolic pathway via liver and kidney. The highest tracer uptake was in thalamus (4.26 ± 0.89) % ID/g and striatum (4.05 ± 1.08) % ID/g at 5 min after injection, followed by cerebral cortex (2.63±0.89) %ID/g, pons (2.26 ±0.57) % ID/g, hippocampus (2. 17 ±0.55) %ID/g and cerebellum (2. 15 ±0.39) %ID/g. Conclusions The automatic synthesis of 11C-CFN is fast and reliable, and this radioligand can be used for opiate receptor imaging.

Objective To investigate the glucose metabolic pattern of brain functional loop in patients with refractory obsessive compulsive disorder (OCD) using 18 F-FDG PET.Methods Eight patients with refractory OCD and 8 age- and gender-matched healthy volunteers underwent 18F-FDG PET brain imaging.SPM software was used for image post-processing and quantitative analysis.Correlation analysis between 18F-FDG uptake and Yale-Brown obsessive compulsive scale(Y-BOCS) score was performed.Results Compared with the controls,the glucose metabolism of bilateral frontal cortices ( including the rectal gyrus,orbital gyrus and cingulate gyrus),left thalamus,right temporal lobe and bilateral cerebellum in refractory OCD patients increased significantly ( Zmax =3.45 - 5.80,all P ＜ 0.001 ).Bilateral motor cortices and bilateral parietal lobes (BA7),however,showed decreased glucose metabolism (Zmax =3.44 - 4.46,all P ＜0.001 ).Y-BOCS score was positively correlated with the glucose metabolism of the bilateral anterior cingulate cortex (Zmax =3.77,3.48 and 2.97,all P ＜ 0.01 ).Conclusions There is a characteristic metabolic pattern of increased glucose utilization in the fronto-striato-thalamic loop and decreased glucose utilization in bilateral motor cortices and parietal lobes in patients with OCD.The glucose metabolism in the anterior cingulate cortex might serve as a quantitative parameter for the assessment of the severity of OCD.

OBJECTIVETo investigate the expression of cathepsin B (CatB) in colorectal cancer tissues and serum levels of CatB in patients with colorectal carcinoma and to study the association of CatB expression with lymph node and li ver metastasis.

METHODSImmunohistochemistry was used to detect CatB expression in tissues, and enzyme linked immunosorbent assay was applied to test CatB levels in peripheral vein blood in 83 patients with colorectal cancer.

RESULTSThe expression rates of CatB in primary lesions, normal colon mucosa, lymph node metastases and hepatic metastases were 56.6%, 31.3%, 88.4%, 85.0% respectively. The positive rates of CatB in primary lesions, hepatic and lymph node metastases were higher than that in normal mucosa (chi (2)=45.6124, P< 0.01). The CatB expression rates in lymph node and hepatic metastases were higher than that in primary lesions chi (2)=11.5982, 4.3747, P< 0.05). The positive rate of CatB was higher in Dukes C and D tumors than that in Dukes A and B tumors (chi (2)=18.8871, 25.1650, P< 0.01), higher in poorly differentiated and mucous adenocarcinomas than that in well-moderately differentiated adenocarcinomas chi (2)=14.2338, P< 0.05). The mean serum level of CatB in 83 patients with colorectal cancer was (5.9+/- 2.9) ng/ml, higher than (2.3+/- 1.1) ng/ml in the controls of 30 healthy volunteers (t=6.6975, P< 0.01). The serum level of CatB in the patients with Dukes C, D stages were higher than that with Dukes A, B stages.

CONCLUSIONSEnhanced expression of CatB in colorectal cancer tissues is associated with tumor infiltration and metastasis. Monitoring serum CatB level in patients with colorectal cancer is important in the prediction and diagnosis of lymph node and hepatic metastasis,and valuable for evaluation of the therapeutic effect.

Adult ; Aged ; Cathepsin B ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging

OBJECTIVETo study the expression of human myofibrillogenesis regulator 1 (MR-1) gene in E. coli and obtain the MR-1 protein and its antibody for further investigation of its biological function.

METHODSExpression vectors pGEX-5X-1, pET30a (+), and pET24a (+), as well as host strain E. coli BL21 (DE3) and BL21-CodonPlus (DE3) -RIL were used for expression of MR-1. MR-1 N-terminal with GST or T7-tag or C-terminal with His-tag, separately, or N terminal with T7-tag and C terminal with His-tag, simultaneously, were fused in plasmids pGEX-5X-1, pET30a (+) , and pET24a (+). The expressed MR-1-T protein, separated and purified by preparative SDS-PAGE, was applied to immunize the rabbits. The titer of the antibody was assayed by ELISA and its immunogenicity was tested by Western blot with pcDNA3/MR-1 transfected human breast cancer cell MCF7.

RESULTSThe MR-1 protein was successfully expressed as inclusion body by fusing its N-terminal with T7-tag in E. coli BL21-CodonPlus (DE3) -RIL. MR-1 protein was purified by electro-elution from SDS-PAGE gel. Using this purified protein, polyclonal antibody in rabbit against MR-1 was essentially generated. ELISA and Western blot showed the titer of this antibody was about 1:10(5) with high immunogenicity.

CONCLUSIONSThe N-terminal fusion tag is the most important mechanism for MR-1 expression. The polyclonal antibody of the generated MR-1 protein in E. coli may be applied for its further biological function studies.

Animals ; Antibodies ; analysis ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Escherichia coli ; metabolism ; Female ; Genetic Vectors ; Humans ; Immunization ; Muscle Proteins ; biosynthesis ; genetics ; immunology ; Plasmids ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Transfection

Adult ; Female ; Follow-Up Studies ; Hepatitis B Surface Antigens ; blood ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Tissue Donors

The present study was to investigate the effect of Salvia miltiorrhiza Bunge. f. alba (SMA) pharmacological pretreatment on apoptosis of cultured hippocampal neurons from neonate rats under oxygen-glucose deprivation (OGD). Cultured hippocampal neurons were randomly divided into five groups (n = 6): normal plasma group, low dose SMA plasma (2.5%) group, middle dose SMA plasma (5%) group, high dose SMA plasma (10%) group and control group. The hippocampal neurons were cultured and treated with plasma from adult Wistar rats intragastrically administered with saline or aqueous extract of SMA. The apoptosis of neurons was induced by glucose-free Earle's solution containing 1 mmol/L Na2S2O4 and labeled by MTT and Annexin V/PI double staining. Moreover, protein expressions of Bcl-2 and Bax were detected by immunofluorescence. The results showed that few apoptotic cells were observed in control group, whereas the number of apoptotic cells was greatly increased in normal plasma group and low dose SMA plasma group. Both middle and high dose SMA plasma could protect cultured hippocampal neurons from apoptosis induced by OGD (P < 0.05). The protective effect of high dose SMA plasma was stronger than that of middle one (P < 0.05). Compared to control, normal plasma and low dose SMA plasma groups, middle and high dose SMA plasma groups both showed significantly higher levels of Bcl-2 (P < 0.05 or 0.01), whereas expressions of Bax was opposite. There were no significant differences of Bcl-2 and Bax expressions between middle and high dose SMA plasma groups. Number of Bcl-2- and Bax-positive cells had similar tendency. Bcl-2/Bax (number of positive cells) ratio was higher in high dose SMA plasma group than those of all the other groups (P < 0.05 or 0.01). These results suggest that pharmacological pretreatment of blood plasma containing middle and high dose SMA could raise viability and inhibit apoptosis of OGD-injured hippocampal neurons by up-regulating the expression of Bcl-2 and down-regulating the expression of Bax.
Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Glucose ; metabolism ; Hippocampus ; cytology ; Ischemic Preconditioning ; methods ; Male ; Neurons ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Salvia miltiorrhiza ; chemistry ; bcl-2-Associated X Protein ; metabolism