AIM: To investigate the role of retinal pigment epithelium (RPE) in the growth of Müller cells using a co-culture system in vitro . METHODS: Müller cells were cocultured with RPE cells under both normoxic and hypoxic conditions in Transwell chamber culture system. Müller cell proliferation was evaluated by MTT assay. The number of cells which migrate through micropores and stay on the outer bottom side of insert systems were observed and counted. RESULTS: The activities of proliferation and migration of Müller cells when cocultured with RPE cells were significantly higher than those of the Müller cells when cultured alone at all time points under both normoxic and hypoxic conditions. However, for both the coculture and control groups, there is no significant difference between the measurements at 3 and 6 hours. CONCLUSION: Evidence suggests that RPE, when co-cultured with Müller cells, can stimulate migration and proliferation of Müller cells under both hypoxic and normoxic conditions in a time-dependent manner; how-ever, there is no evidence to support the synergetic interaction of RPE and Müller cells co-cultured under hypoxic conditions.

Background More and more evidences suggest that the interaction of retinal progression of retinal diseases.Objective Present study was to investigate the primarily cultured and digested using explant culture method and trypsas acidic protein(GFAP) staining,S-100 staining and cytokine-18 staining co-cultured under the normoxia and hypoxia using transwell chamber,and the proliferation and migration of cultured RPE cells were examined using MTT and compared with only RPE cells cultured group at 3, 6, 24 and 48 hours.Results Over 90% of primarily cultured RPE cells showed the positive response for S-100.The proliferation and migration of RPE cells were significantly increased in hypoxia condition compared with normoxia condition (P＜0.01).In hypoxia group,amount of proliferation and migration of RPE cells in co-culture group were higher than the only RPE cells cultured group(P＜0.01).Conclusion Hypoxia appear to aggravate the proliferation and migration of RPE cells under the hypoxia status.

Emerging evidence has indicated that circular RNAs (circRNAs) play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors.However,the clinical role of circRNAs in bladder cancer (BC) and the molecular mechanisms have yet to be fully understood.In this study,the clinical specimens were obtained and the expression level of a circRNA BCRC4 was detected by real-time PCR in both BC tissues and cell line.The circular RNA over-expression plasmid was constructed and transfected into BC cells and related cell line.The cell cycles and apoptosis were observed using inverted microscope and flow cytometry.Western blotting was used to compare the relative protein expression of groups with different treatments.It was found that circRNA BCRC4 expression was lower in BC tissues than in adjacent normal tissues.Furthermore,consequences of fomed-expression of BCRC4 promoted apoptosis and inhibited viability of T24T and UMUC3 cells,and up-regulated BCRC4-inereased miR-101 level,which suppressed EZH2 expression in both RNA and protein levels.In addition,gambogic acid (GA) is a promising natural anticancer compound for BC therapy,and GA treatment increased the BCRC4 expression in T24T and UMUC3 cells in a dose-dependent manner.Altogether,our findings suggest that BCRC4 functions as a tumor suppressor in BC,and mediates anticancer function,at least in part,by up-regulating the expression of miR-101.Targeting this newly identified circRNA may help us develop a novel strategy for treating human BC.

OBJECTIVETo observe the inhibiting effect of acupuncture on blood lipid, myocardial hypertrophy and fibrosis in mice with hyperlipemia, and explore its possible action mechanism.

METHODSTen inbred mice (C57) were applied. Forty ApoE(-/-) mice who removed gene of apolipoprotein E were randomly divided into a control group, a non-acupoint group, an acupoint group and a medication group. The points 0. 5 cm and 1 cm next to the end of mice tail were respectively punctured in the non-acupoint group; "Neiguan" (PC 6) and "Fenglong" (ST 40) were punctured in the acupoint group; intragastric administration of simvastatin was applied in the medication group. After 8 weeks of treatment, the changes of total cholesterol (TC) and ratio of heart to body mass in each group were measured; changes of cardiac muscle fiber and ventricular wall thickness were observed; enzyme linked immunosorbent assay (ELISA) was used to test the level of angiotensin II (Ang I ) in plasma, and western blotting method was used to test protein content of angiotensin II type 1 receptor (AT1R) and endothelin-1 type A receptor (ETAR) in the heart.

RESULTSAfter 8 weeks of intervention, compared with the control group, rising range of blood lipid was obviously decreased (P<0.01) in the acupoint group and medication group, ratio of P<0.01), myocardial heart to body mass was decreased (P<0.05), thickness of ventricular wall was reduced (P fibrosis was relieved, levels of Ang II and ET-1 in plasma were decreased (P<0. 05), content of NO was increased (P<0. 05), and protein content of AT1R and ETAR was decreased in the heart (P<0. 05).

CONCLUSION40) could inhibit the rising of blood lipid in ApoE(-/-) mice, lower the levels of Ang II and ET-1 in peripheral blood, increase the content of NO and inhibit the expression of AT1R and ETAR in heart tissue, which could relieve myocardial hypertrophy and fibrosis to play a protective role on heart.

Acupuncture Points ; Acupuncture Therapy ; Angiotensin II ; metabolism ; Animals ; Blood Pressure ; Disease Models, Animal ; Heart ; physiopathology ; Heart Diseases ; etiology ; metabolism ; physiopathology ; prevention & control ; Humans ; Hyperlipidemias ; complications ; physiopathology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Myocardium ; metabolism

Hernia, Inguinal ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Peritoneal Dialysis, Continuous Ambulatory ; Peritoneum ; diagnostic imaging ; Tomography, X-Ray Computed ; methods

Objective To study the changes of renal cortex angiotensin Ⅱ (AngⅡ) expression and podocyte autophagy in aging rats with normotension versus hypertension,to further investigate the possible mechanism of renal injury in hypertension during aging.Methods Normotensive Wistar-Kyoto-rats (WKYs) were allocated to three groups by age of 3-month-old group,13-month-old group,22-month-old group.Gender,age and number-matched spontaneously hypertensive rats (SHRs) served as controls.Blood pressure was monitored.Levels of urinary albumin,urinary creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),Ang Ⅱ in serum and in renal cortex were detected.The kidney ultrastructural changes and podocyte autophagosomes were observed under light and transmission electron microscopy.Expressions of nephrin,LC3B Ⅱ,Atg5 and p62 in glomeruli were analyzed by Western blotting.Results Blood pressure was significantly higher in SHRs than in WKYs (P＜0.05).During aging from 3-month-old to 13-month-old and to 22 monthold group,the urinary albumin/creatinine ratio was increased significantly in SHRs with hypertension and in WKYs with normotension [WKY:(0.12±0.01) g/mmol,(0.14±0.04) g/mmol vs.(0.34± 0.05) g/mmol;in SHR:(0.29±0.04) g/mmol,(0.31±0.05) g/mmol vs.(0.72±0.16) g/mmol,P＜0.05],but SCR and BUN levels had no significant difference in SHRs and WKYs during aging (both P＞0.05).And Ang Ⅱ levels in both serum and renal cortex had no significant change in normotension group during aging three time points (both P＞0.05).An age-dependent decrease of Ang Ⅱ level in renal cortex appeared during aging three time points in hypertensive rats [SHR:(0.02 ±0.00) μg/L,(0.02±0.00) μg/L vs.(0.01±0.00) μg/L,P＜0.05],but serum level of Ang Ⅱ had no significant difference during aging three time points (P＞ 0.05) in hypertension group.The structural changes,including glomerulosclerosis,tubular atrophy and interstitial fibrosis were observed during aging three time points both in normotensive and hypertensive rats,but these pathological changes were more serious in hypertensive rats.Autophagosomes relatively accumulated in aging normotensive rats.The protein expressions of LC3B Ⅱ,Atg5 and p62 were increased in normotensive rats during aging (all P＜0.05).While the protein expressions of nephrin,LC3B Ⅱ,Atg5 and p62 were decreased in aging hypertensive rats (all P ＜ 0.05).Conclusions Ang Ⅱ expression level in renal cortex is decreased during aging in hypertensive rats.Podocyte autophagic activity is relatively decreased during aging in normotensive rats,but is relatively increased during aging in hypertensive rats.Ang Ⅱ-induced podocyte autophagy may be involved in the pathogenesis of renal injury in hypertension during aging.

Objective To examine the effects of benazepril and losartan on glomerular podocyte autophagy in aged spontaneously hypertensive rats (SHRs) and investigate the underlying mechanisms of renal-protective effects of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin receptor blockers (ARB).Methods Wistar-Kyoto rats (WKYs) were used as the normal control (NORM) group (2 ml physiological saline per day).SHRs were randomly divided into 4 groups:the CTRL group (2 ml physiological saline per day),the ACEI group (10 mg · kg-1 · d-1),the ARB group (30 mg· kg-1 ·d-1) and the combined group (10 mg· kg-1 · d-1 benazepril and 30 mg· kg-1 · d-1),with six 18-month-old make rats in each group.The experiments were conducted during a 4-month period.Blood pressure was monitored regularly.At the end of the experiments,we measured the levels of urine protein,urine creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),and serum and renal cortex angiotensin Ⅱ (AngⅡ).Ultrastructural changes in the kidney were examined under light and transmission electron microscopy.The expressions of nephrin,LC3BⅡ,Atg5 and p62 in the glomerulus were analyzed by Western blot analysis.Results After treatment,the blood pressure and the urine albumin/creatinine ratio of the four SHR groups were still significantly higher than those of the NORM group,but the blood pressure and the urine albumin/creatinine ratio of the ARB group and the combined group were significantly lower than those of the CTRL group (all P＜ 0.05);There were no significant differences in SCR and BUN levels among these five groups (P＞ 0.05);The level of serum AngⅡ of the combined group was significantly higher than that of the CTRL group [CTRL (0.08±0.00) μg/L,Combined (0.12±0.01) μg/L,P＜0.05];The levels of cortex AngⅡ of the four SHR groups were significantly lower than those of the NORM group,while the level of cortex AngⅡ of the ARB group was significantly higher than that of the CTRL group (all P＜0.05);Renal ultrastructural examination revealed shrunken glomeruli,fused or effaced epithelial cell foot processes,and focal atrophy of renal tubules in the four SHR groups.These pathological changes were more serious in the CTRL group but less so in the combined group.There were significantly more autophagosomes in the NORM group and the combined group than in the CTRL group (P＜0.05).Compared with the NORM group,the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the CTRL group were suppressed significantly (P ＜ 0.05).The expressions of nephrin,LC3BⅡ and Atg5 in the ACEI group and the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the ARB group and the combined group were higher than in the CTRL group (P＜0.05).Conclusions ACEI/ARB can decrease the autophagic activity of glomerular podocytes.The renal-protective effects of ACEI/ARB may be mediated by glomerular podocyte autophagy,which is induced by AngⅡ.

OBJECTIVETo investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.

METHODSRat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.

RESULTSThe majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).

CONCLUSIONSalidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.

Animals ; Apoptosis ; drug effects ; physiology ; Caspase 3 ; metabolism ; Cell Hypoxia ; physiology ; Cells, Cultured ; Glucosides ; pharmacology ; Humans ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Penis ; cytology ; drug effects ; Phenols ; pharmacology ; Rats

OBJECTIVETo observe the clinical effect of low-dose once-daily tadalafil combined with Shuganyiyang Capsules in the treatment of mild-to-moderate erectile dysfunction (ED).

METHODSNinety patients with mild-to-moderate ED were equally randomized to groups A, B and C to receive Shuganyiyang Capsules, tadalafil, and tadalafil + Shuganyiyang Capsules, respectively. The scores of the patients on IIEF-5 and SF-PAIRS (15-Item Short Form of Psychological Interpersonal Relationship Scales) were recorded before and at 1 and 3 months after treatment.

RESULTSThe IIEF-5 scores of groups A, B and C were 10.13 +/- 1.55, 11.00 + 1.60 and 10.73 +/- 1.91 before treatment, and 13.77 +/- 2.11, 17.77 +/- 2.13 and 17.17 +/- 3.84 at 1 month after treatment, significantly higher in B and C than in A (P <0. 001) , but with no remarkable difference between B and C (P =0. 411). At 3 months after treatment, the IIEF-5 scores were 15.77 +/- 2.05, 18.07 +/- 2.24 and 19.37 +/- 3.76 in the three groups, dramatically higher in B and C than in A (P <0.001) as well as in C than in B (P<0.05). The scores on sexual self-confidence, sexual spontaneity and time concerns in SF-PAIRS were 3.90 +/-0.80, 8.67 +/- 1.94 and 14.43 +/- 1.92 before medication, 5.83 +/- 1.02, 9.90 +/- 1.75 and 11.17 +/- 1.68 at 1 month and 6.73 +/- 0.98, 11.07 +/- 2.08 and 10.67 +/-1.60 at 3 months after medication in group A; 4.17 +/- 0.87, 9.37 +/-1.43 and 14.47 +/-1.57 before medication, 6.47 +/-0.78, 10.83 +/- 2.18 and 10.20 +/-1.56 at 1 month and 6.83 +/-0.91, 11.30 +/- 1.88 and 9.47 +/- 1.57 at 3 months in group B; and 4.23 +/-0. 94, 9.50 +/- 1.89 and 14.67 +/- 2.91 before medication, 8.03 +/- 1.67, 13.43 +/-1.10 and 9.70 +/-1.21 at 1 month and 8.93 +/- 1.78, 14.70 +/- 1.26 and 8. 87 +/- 0. 97 at 3 months in group C. Compared with the baseline, the SF-PAIRS scores of the three groups were all significantly improved after treatment (P <0. 05) , and markedly higher in C than in the other two groups (P <0.05).

CONCLUSIONLow-dose once-daily tadalafil combined with Shuganyiyang Capsules is obviously effective in the treatment of mild-to-moderate ED, which not only improves the patients'erectile function, sexual self-confidence and sexual spontaneity, but also reduces their time concerns.

Adult ; Carbolines ; administration & dosage ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Tadalafil ; Treatment Outcome

Objective To construct a system for three dimensional face scanning and measurement. Methods The measurement system was based on the principle of triangulation and the combination of gray-code and phase-shift structured light projection. The system software was developed for Windows XP with the aid of tools such as Visual C++ and Hoops. Results A three dimensional measurement system based on structured light projection was developed. The system hardware was composed of fringe projection unit, image gathering unit, system control unit and mechanical appearance, and the system software was composed of point cloud display and editing module. The lamp house of the system was 12V, the working distance was 900 mm, the scanning time was 5.5 s and the scanning field was 500 mm×400 mm. Conclusion The three dimensional measurement system based on structured light projection is a refined machine with safe light to eyes, and the accuracy and scanning speed are suitable to face scanning.