OBJECTIVETo investigate the expressions of sphingosine-1-phosphate receptors 1-3 (S1P1- 3) in the corpus cavernosum of castrated male rats and its relationship with the NOS/NO/cGMP and RhoA/Rho kinase signaling pathways.

METHODSWe equally randomized 18 eight-week-old healthy male SD rats into a sham-operation control, a castration, and a testosterone replacement (TR) group and harvested the bilateral testes and epididymides from the rats in the latter two groups, followed by 4 weeks of subcutaneous injection of testosterone propionate at 3 mg per kilogram of the body weight per day for those in the TR group and that of plant oil for those in the control and castration groups. At the age of 12 weeks, we measured the serum testosterone (T) level and maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP) of the animals and determined the expressions of SlP1-3, eNOS, P-eNOS, ROCK1, and ROCK2 in the corpus cavernosum by Western blot and immunohistochemistry.

RESULTSThe serum T level was significantly decreased in the rats of the castration group as compared with those of the control and TR groups ([0.41 ± 0.04] vs [16.01 ± 1.02] and [15.84 ± 1.32] nmol/L, P < 0.01), with no statistically significant difference between the latter two groups. The ICPmax/MAP at 0 V, 3 V, and 5 V electric stimulation was remarkably lower in the rats of the castration group (0.088 ± 0.014, 0.323 ± 0.014, and 0.432 ± 0.012) than in those of the control group (0.155 ± 0.011, 0.711 ± 0. 010, and 0.819 ± 0.024) and TR group (0.153 ± 0.012, 0.696 ± 0.017, and 0.763 ± 0.027) (P < 0.01), with no significant difference between the latter two groups. With GAPDH as internal control, the animals of the castration group showed markedly reduced expressions of S1P1 ([49.99 ± 3.39]%), eNOS ([46.82 ± 3.81]%) , and P-eNOS ([45.42 ± 4.35]%) in comparison with those in the control group ([72.57 ± 3.06], [89.76 ± 3.98], and [82.53 ± 8.92] and TR group ([71.77 ± 4.43], [87.19 ± 4.23], and [79.82 ± 7.38]%) (P < 0.01) , while the expressions of S1P2, S1P3, ROCK1, and ROCK2 were significantly upregulated in the castration group ([82.35 ± 4.13], [61.03 ± 5.14], [74.50 ± 4.02], and [69.83 ± 5.75]%) as compared with those in the control group ([41.67 ± 1.68], [31.66 ± 2.67], [35.69 ± 5.56], and [39.85 ± 7.17]%) and TR group ([42.80 ± 3.87], [32.25 ± 4.22], 38.06 ± 5.21], and [42.36 ± 4.44]%) (P < 0.01).

CONCLUSIONAndrogen deficiency induces significant reduction of ICPmax/ MAP in male rats, which is possibly associated with the decline of S1P1 in the corpus cavernosum, inhibition of the eNOS/NO/cGMP signaling pathway, increased expressions of S1P2 and S1P3, and activation of the RhoA/Rho kinase signaling pathway.

Animals ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Orchiectomy ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Lysosphingolipid ; metabolism ; Testosterone ; blood ; pharmacology ; rho-Associated Kinases ; metabolism

AIM: To screen the lentiviral vector carrying siRNA with higher efficiency of suppressing the sphingosine-1-phosphate receptor 2 (S1P2) gene expression in the primarily cultured corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).METHODS: SHR and SD rats (n=5 each) were used for primarily culturing corpus cavernosum smooth muscle cells.The cells were randomly divided into 6 groups: SHR siRNA-1, SHR siRNA-2, SHR siRNA-3, SHR GFP, SHR control (SHR non-transfection group), and SD control (SD rat control group).Each group had 5 samples with 1.0×105 cells of each sample.At 72 h after transfection (MOI=60) with lentiviral vectors carrying S1P2 siRNA into the SHR corpus cavernosum smooth muscle cells, the expression of GFP was observed under fluorescence microscope.The protein expression of S1P2, ROCK1, ROCK2 and eNOS in the corpus cavernosum smooth muscle cells, and the mRNA expression of S1P2, ROCK1 and ROCK2 were determined by by Western blot and RT-PCR.RESULTS: The transfection efficiency of the corpus cavernosum smooth muscle cells in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SHR GFP groups were>80%.Compared with SHR control group, the mRNA levels and the protein expression of S1P2, ROCK1 and ROCK2 in SHR GFP group showed no remarkable changes, while those in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SD control groups were significantly lower than those in SHR control group (P<0.05).The protein expression of eNOS in SHR siRNA-1, SHR siRNA-2, SHR siRNA-3 and SHR GFP groups were not significantly changed as compared with SHR control group, but that in SD control group was significantly higher than that in SHR control group.CONCLUSION: Three groups of siRNA lentiviral vectors targeting S1P2 inhibit the expression of S1P2 in the corpus cavernosum smooth muscle cells of SHR, and by silencing the S1P2 expression, the expression of ROCK1 and ROCK2 is inhibited.Among them, siRNA-1 has the highest inhibitory efficiency.

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

This study examined the mRNA expression of NALP3 in the spleen of the mice with hypersplenism due to portal hypertension (PH). The mouse hypersplenism models were established by oral administration of tetrachloromethane (2 mL/kg/week for 12 weeks by oral gavage). All the mice were randomly divided into a control group and an experimental group. The blood routine test was conducted, spleen index was calculated and spleen was histologically examined. Portal vein sera were taken for detection of the level of uric acid. The mRNA expressions of NALP3 and IL-1beta in the spleen were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that the platelet count was significantly lower in the experimental group [(674+/-102)x10(9)/L] than in the control group [(1307+/-181)x10(9)/L] (P<0.05), while the spleen index was significantly higher [(9.83+/-1.36) mug/g] in the experimental group than in the control group [(4.11+/-0.47) mug/g] (P<0.05). The histopathological changes of spleen followed the pattern of congestive splenomegaly. No significant difference was found in the uric acid level in the portal vein between the control group and the experiment group. The mRNA expressions of NALP3 and IL-1beta were up-regulated significantly in the spleen in the experimental group as compared with those in the control group (P<0.05). It was concluded that NALP3 and IL-1beta may play important roles in the pathogenesis of hypersplenism.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Transfusion, Autologous ; methods ; Female ; Humans ; Male ; Middle Aged ; Orthopedics ; Postoperative Complications ; Young Adult

OBJECTIVETo study the status of teeth loss and their restorations of the middle-aged and elders in Liaoning province.

METHODSInvestigated the tooth losing and their restoration of middle-aged (35-44 years old) and elders (65-74 years old) according the method of the third national epidemiological investigation of oral health. The data was statistically analyzed by SPSS 11.0 software.

RESULTSOf all 1,584 the subjects, the sum of tooth loss were 6,776, mean tooth loss were 4.28. The rate of completion of dentition was 37.44%. There were 104 middle-aged and elders who happened deletion of dentition. There were 887 middle-aged and elders who happened defection of dentition. The tooth loss of maxillary and mandible was 3,550 and 3,226 respectively. The rate of prosthetic restoration was 52.77%.

CONCLUSIONThe oral health status of middle-aged and elders should be taken vey importance, especially those who lived in rural area. Dental caries and periodontal disease should be prevented so that to promote the status of middle-aged and elders oral health and to improve the quality of life.

Dental Caries ; Dentition ; Female ; Humans ; Male ; Middle Aged ; Oral Health ; Periodontal Diseases ; Quality of Life ; Tooth Loss

OBJECTIVE: To investigate the regulation of hepatitis C virus (HCV) core protein on the activity of signal transducers and activators of transcription 3 (stat3). METHODS: A cell line expressing stable HCV core protein-QSG7701-core was constructed by transfecting the pcDNA3. 1-core (expressing HCV core protein) into the human immortalized hepatocyte line QSG7701. The phosphorylation and DNA binding activity of stat3 were detected by immunocytochemistry, Western blot, and electrophoretic mobility shift assay (EMSA). RESULTS: The expression level of phosphorylated stat3 in QSG7701-core cells was significantly lower than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells, but there were no significant differences in the expression levels of total stat3 among the 3 groups. The positive signal of phosphorylated stat3 in nucleus of QSG7701-core cells was obviously weaker than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells. EMSA showed that DNA binding activity of stat3 in QSG7701-core cells significantly decreased. CONCLUSION The expressionof HCV core protein in human hepatocyte line may suppress the phosphorylation and DNA binding activity of stat3, which may be one of the causes for resistance against interferon.
Cell Line ; DNA-Binding Proteins ; metabolism ; Hepatocytes ; cytology ; metabolism ; Humans ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Transfection ; Viral Core Proteins ; biosynthesis ; genetics

OBJECTIVETo study the dental caries status of the 65-74 years old people in northeast of China, analyze the distribution of caries in the group of people, and provide scientific basis for establishing the oral health care policies.

METHODSThe representative samples were selected by multi-stage, stratified and random sampling from the three province of northeast of China. Dental caries of 2 364 people aged 65-74 were recorded according to WHO criteria. SPSS 12.0 software was used to analyze the data.

RESULTS1) Among 2,364 elderly people in northeast of China, the prevalence of dental caries was 67.5% and DFT was 2.68, the composition of filling was 10.82%. 2) The prevalence of coronal caries was 66.4% and DFT was 2.55. The composition of filling was 11.37%. 3) The prevalence of root surface caries was 48.4% and DFT was 1.64. The composition of filling was only 1.8%.

CONCLUSIONThe prevalence of dental caries among elderly people in northeast of China is still very high, and the filling rate is low. The community based oral health promotion should be strengthened and preventive-oriented oral health care systems are needed in northeast of China.

Aged ; China ; Dental Caries ; Humans ; Prevalence ; Risk Factors ; Root Caries

OBJECTIVETo survey the tooth decay of the first molars among the people in area of northeast China in order to provide the foundation for prevention and treatment strategies of caries.

METHODSAccording to the third national oral health investigation of epidemiology, the dental caries of first permanent molars were examined among 9394 people aged 5, 12, 35-44 and 65-74 in northeast China. The data were analyzed by SPSS 13.0 statistical software. Dental caries prevalence rate and DMFT (dmft) of first permanent molar were calculated.

RESULTSThe prevalence of crown and root dental caries in northeast China were 26.86% and 3.81% respectively, mean DMFT (dmft) of crown and root dental caries were 0.41 and 0.04 respectively. There were significant differences of dental caries among provinces in northeast China. Mandibular first molar suffered from dental caries easily. The prevalence of caries of female was higher than that of male (P<0.01), but there was not significant difference of prevalence of root caries between two sexes (P>0.05). Except Jilin province, there were significant differences of prevalence of crown caries between city and rural area in Liaoning and Heilongjiang provinces (P<0.01). It was no significant difference of prevalence and mean DMFT (dmft) of root caries between city and rural area in each province (P>0.05).

CONCLUSIONThe prevalence of dental caries of the first molar among the people in northeast China is high. The dental caries prevention and cure of first permanent molar should be emphasized in oral health field.

China ; Dental Caries ; Female ; Humans ; Male ; Molar ; Oral Health ; Prevalence ; Root Caries

OBJECTIVETo study cardiac troponin T (TNNT2) gene mutations in Chinese patients with hypertrophic cardiomyopathy (HCM) and analyze the correlation between the genotype and phenotype.

METHODSNinety-five unrelated Chinese patients with HCM and 120 control individuals were screened for TNNT2 gene mutations. Seven exons (8, 9, 10, 11, 14, 15, and 16) in the functional regions of TNNT2 gene were amplified using PCR and the products were sequenced. The patients with positive results underwent further family screening.

RESULTS AND CONCLUSIONThis study did not find any HCM-caused mutations in TNNT2 gene, a result different from the reported rates of TNNT2 gene mutation ranging from 10% to 20% in other nations, suggesting that TNNT2 gene is not a susceptible gene for HCM in Chinese population.

Asian Continental Ancestry Group ; genetics ; Cardiomyopathy, Hypertrophic ; genetics ; Case-Control Studies ; Humans ; Mutation ; Troponin T ; genetics