OBJECTIVE: To investigate the development of the enteric nervous system(ENS) in the middle stage(the 4th approximate, equals 6th months) of human embryogenesis. METHODS: A histologic evaluation of the clonic enteric nervous system was done using NADPHd histochemistry, PAP immunohistochemistry, with anti-PGP 9.5 and anti-S-100 protein. RESULTS: During this stage of embryology three things were noted. (1)The nerves in the myenteric layer increased markedly. This was shown by the PGP 9.5 immunoreactive nerves spreading out and the S-100 protein immunoreactive nerves forming a "bamboo basket"pattern. (2)The whole myenteric colon showed nitrigeric nerves paralleling the growth of the myofibers in the circular muscle layer. Nitrigeric perikara were rarely found in the extrinsic submucosal layer.(3) In the whole-mounted preparations reactive nerves formed the complex nerve net in the myenteric layer. CONCLUSION: The middle stage of embryogensis is very important to the development of the colon ENS.

Candida shehatae xyl1 gene and Pichia stipitis xyl2 gene were amplified by PCR and the xyl1 and xyl2 were both placed under the promoter GAL of vector pYES2 to produce the recombinant expression vector pYES2-P12. Subsequently the pYES2-P12 vector was transformed into S. cerevisiae YS58 by LiAc to produce the recombinant yeast YSS8-12. It was indicate that the recombinant yeast YSS8-12 could converse xylose to ethanol with the xylose consumption rate of 81. 3%.

Objective To detect the impairment of response inhibition and working memory in patients with alcohol dependence.Methods A total of forty-eight alcohol dependent patients and fifty age,gender,IQ,education matched controls were recruited.Neuropsychological tests were applied to measure the differences of response inhibition and working memory between the two groups.Results In the response inhibition task,the patient group had more commission errors ((7.02± 3.48) vs (3.45± 1.52)) and longer reaction time ((605.45 ± 142.56)ms vs (435.72±51.18)ms) compared to the control group (t=6.534,P=0.000; t=7.781,P=0.000).In the spacial working memory task,the patient group also had more commission errors ((4.58± 3.45) vs (0.43± 0.88)) and longer reaction time((10566.16±2455.61) ms vs (9185.44±2677.52) ms) than control (t=8.085,P =0.000; t=2.657,P=0.009).Conclusion There are significant deficiencies in response inhibition and working memory in patients with alcohol dependence.

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

OBJECTIVETo investigate the role of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) pathway in erectile dysfunction (ED) caused by the absence of testosterone (T).

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into groups A (control) , B (castration), and C (castration + androgen replacement). The rats in groups B and C were castrated surgically, and those in C injected with T undecanoate (100 mg/kg) at 1 week after castration, while the others with 0.9% normal saline instead. At 1 month after treatment, we determined the serum T level, intracavernous pressure (ICP), and mean carotid arterial pressure (MAP) of the rats, and detected the expressions of ERK1/2 and endothelial nitric oxide synthase (eNOS) by Western blot.

RESULTSThe serum T level was significantly lower in group B ([1.27 ± 0.48] nmol/L) than in A ([17.14 ± 1.07] nmol/L) and C ([16.24 ± 1.90] nmol/L) (P < 0.05), and so were ICP and MAP (P < 0.05). The expression of ERK1/2 showed no statistically significant differences among the three groups (P > 0.05), that of phosphatase ERK1/2 was markedly higher while that of eNOS remarkably lower in group B than in A and C (both P < 0.05).

CONCLUSIONAndrogen replacement may improve the erectile function of castrated rats by regulating the ERK1/2 pathway.

Androgens ; therapeutic use ; Animals ; Blotting, Western ; Erectile Dysfunction ; drug therapy ; metabolism ; Hormone Replacement Therapy ; MAP Kinase Signaling System ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Orchiectomy ; Penile Erection ; Penis ; Rats ; Rats, Sprague-Dawley ; Testosterone ; analogs & derivatives ; therapeutic use

Objective To discuss the molecular mechanism of 18F-fluorodeoxyglucose (FDG) uptake in tumor and to assess its value to identify pathologic type and cancer staging in patients with earlystage nasopharyngeal carcinoma.Methods Forty patients with nasopharyngeal carcinoma of early-stage,including 12 cases with T1 stage and 28 cases with T2 stage, underwent FDG PET imaging.The maximum standardized uptake value ( SUVmax ) and mean standardized uptake value ( SUVmean ) of FDG uptake of each patient were measured and compared between T1 and T2 stage by t-test.The expression of glucose transport protein 1 ( Glut1 ) and hexokinase- Ⅱ ( HK- Ⅱ ) of each case was measured in paraffin sections by streptavidin-perosidase (SP) immunohistochemistry.The positive expression rate of Glut1 and HK- Ⅱ was calculated and compared between T1 and T2 by x2 test.Meanwhile, the correlation between the expression of Glut1 or HK-Ⅱ and the SUVmax was tested by Pearson analysis.Results The SUVmax and SUVmean in 40 patients were 9.45 ± 1.87 and 6.04 ± 1.09, respectively.The SUVmax of patients with T1 stage (8.95 ± 1.91 ) was significantly lower (t =4.46, P＜0.001 ) than that of patients with T2 stage (11.55 ± 1.70), and the SUVmean of patients with T1 stage (5.61 ± 1.08) was significantly lower ( t = 6.76, P ＜ 0.001 ) than that of patients with T2 stage (7.98 ± 1.10) too.Among 40 patients, all patients showed positive expression of Glut1 and HK-Ⅱ , and the positive expression rate of Glut1 and HK-Ⅱ was ( 45.2 ± 10.9 )% and ( 68.3 ±9.5)%, respectively.The positive expression rate of Glut1 was (38.4 ±8.1)% in T1 stage and (49.7 ±12.6)% in T2 stage, which displayed no difference (x2 =40.58, P＞0.05), but the HK-Ⅱ positive expression rate showed significant difference (x2 =58.71, P＜0.05) between T1 stage (60.1 ±11.1)% and T2 stage (77.9 ± 14.7 )%.The correlation analysis indicated that there was low-degree positive correlation (r =0.369, P=0.019) between the SUVmax and Glut1 expression, and there was medium-degree positive correlation (r = 0.549, P = 0.001 ) between the SUVmax and HK-Ⅱ expression.Conclusion Expression of Glut1 and HK-Ⅱ was positively correlated with FDG uptake in patients with early-stage nasopharyngeal carcinoma.

OBJECTIVETo optimize the culture conditions for clonal isolation of rat bone marrow-derived multipotential adult progenitor cells (rMAPC) and identify their surface markers and differentiation potentials.

METHODSBy using a low concentration of fetal bovine serum culture medium, rMAPCs were primarily isolated from bone marrow by attachment culture and clonal-like cells were selected by single cell limiting dilution. The surface antigens of the cloned rMAPC were analyzed by flow cytometry and immunocytochemistry. Multi-differentiation capacities were evaluated by lipoblasts and osteoblasts and neuroblasts differentiation induction. The expressions of Oct-4 and three embryonic germ layer markers were detected by RT-PCR.

RESULTSSingle cell-derived rMAPC could be expanded to passage 20 in vitro which still maintained active proliferation ability. The expanded rMAPCs expressed CD71, alpha-SMA and vimentin, but not CD34, CD44 and CD45. About 83% of the rMAPCs was in the resting phase(G0 + G1) of cell cycle and 17% in S + G2 + M phase. They could be induced to differentiate into adipogenic cells, osteogenic cells and neural like cells. RT-PCR demonstrated that there were expressions of oct-4 gene and three embryonic germ layer markers on the rMAPCs.

CONCLUSIONSCloned rMAPC can maintain the phenotypes of stem cell during in vitro culturing. It might be an potential adult stem cell source for therapeutic stem cell transplanting and tissue engineering.

Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Cell Line ; Culture Media, Conditioned ; Flow Cytometry ; Male ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the epidemiological changes of erectile dysfunction (ED) patients in the past five years.

METHODSIn 2003 and 2008, we conducted two questionnaire investigations on the epidemiological changes of ED outpatients in 11 Chinese cities in such aspects as age, disease course, ED severity, smoking and drinking habits, accompanying hypertension, diabetes and coronary heart disease (CHD), and sexual intercourse satisfaction.

RESULTSAccording to the valid copies of the questionnaire collected (808 in 2003 and 858 in 2008), the age pattern of the ED patients hardly changed in the past five years, over 60% aged 30 - 50 years. Compared with the results obtained in 2003, the second investigation showed obvious increases in the following numbers of the ED patients: by 13% in those with longer disease courses (5 - 10 yr), from 24.1 to 42.9% in those with moderate ED, from 20.4 to 29.9% in those with severe ED, by at least 10% in those with smoking and drinking habits, from 11.5 to 16.2% in those with hypertension, from 9.4 to 13.5% in those with diabetes, and from 57.6 to 73.3% in those without sexual satisfaction, while the number of those with CHD did not change significantly.

CONCLUSIONIncreased unhealthy living habits and erectile function impairing diseases have added to the incidence and severity of ED. There is still much work to be done in the prevention and early treatment of ED.

Adult ; China ; epidemiology ; Erectile Dysfunction ; epidemiology ; Humans ; Incidence ; Male ; Middle Aged ; Outpatients ; Penile Erection ; Surveys and Questionnaires ; Urban Population

OBJECTIVETo investigate the DNA damage of human lens epithelial cells (LECs) caused by acute exposure to low-power 217 Hz modulated 1.8 GHz microwave radiation and DNA repair.

METHODSCultured LECs were exposed to 217 Hz modulated 1.8 GHz microwave radiation at SAR (specific absorption rate) of 0, 1, 2, 3 and 4 W/kg for 2 hours in an sXc-1800 incubator and irradiate system. The DNA single strand breaks were detected with comet assay in sham-irradiated cells and irradiated cells incubated for varying periods: 0, 30, 60, 120 and 240 min after irradiation. Images of comets were digitized and analyzed using an Imagine-pro plus software, and the indexes used in this study were tail length (TL) and tail moment (TM).

RESULTSThe difference in DNA-breaks between the exposure and sham exposure groups induced by 1 and 2 W/kg irradiation was not significant at every detect time (P > 0.05). As for the dosage of 3 and 4 W/kg there was difference in both group immediately after irradiation (P < 0.01). At the time of 30 min after irradiation the difference went on at both group (P < 0.01). However, the difference disappeared after one hour's incubation in 3 W/kg group (P > 0.05), and existed in 4 W/kg group.

CONCLUSIONNo or repairable DNA damage was observed after 2 hour irradiation of 1.8 GHz microwave on LECs when SAR < or = 3 W/kg. The DNA damages caused by 4 W/kg irradiation were irreversible.

Cell Phone ; Cells, Cultured ; Comet Assay ; DNA Damage ; radiation effects ; DNA Repair ; Dose-Response Relationship, Radiation ; Epithelial Cells ; radiation effects ; Humans ; Lens, Crystalline ; cytology ; radiation effects ; Microwaves

OBJECTIVEA reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of HIV-1.

METHODSRT-LAMP primers were designed according to conservative sequences of HIV-1 gag gene, and their sensitivity and specificity were evaluated by the established RT-LAMP protocol with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The performance of RT-LAMP on clinical samples was compared with real-time reverse transcription PCR(qRT-PCR).

RESULTSThe RT-LAMP assay showed a high specificity, and its detection limit was 1000 copies RNA per tube. The sensitivity and specificity of this method using 43 clinical samples were 94.6% and 100%, respectively,in comparison with those of qRT-PCR.

CONCLUSIONRT-LAMP assay using hydroxynaphthol blue dye does not need expensive instruments, and offer an alternative for the rapid detection of HIV-1 with the potential to be applied in field diagnosis.

HIV-1 ; isolation & purification ; Naphthalenesulfonates ; Nucleic Acid Amplification Techniques ; methods ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Reverse Transcription ; Sensitivity and Specificity