AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.

OBJECTIVETo analyze the parameters of urodynamic tests for patients with type-III B prostatitis and evaluate the significance of the results of urodynamic tests in the choice of therapies for this disease.

METHODSUrodynamic tests were performed for 87 type-III B prostatitis patients aged 22-45 (30.7 ± 8.5) years, who had moderate or severe lower urinary tract symptoms (LUTS) and failed to respond to routine therapy. Different treatments were administered according to the results of urodynamic tests followed by observation of the therapeutic effects.

RESULTSUrodynamic abnormalities were found in 70 of the 87 patients, bladder outlet obstruction in 28 (32.2%), detrusor overactivity in 25 (28.7%), bladder hyperesthesia in 18 (20.7%), low compliance in 10 (11.5%), detrusor-external urethral sphincter dyssynergia in 1 (1.1%), and impaired detrusor contractile function in 1 (1.1%). Treatments achieved obvious effectiveness in 26 cases (29.9%), effectiveness in 51 (58.6%), and no effectiveness in 10 (11.5%).

CONCLUSIONUrodynamic tests contribute significantly to the choice of therapies for type-III B prostatitis patients with moderate or severe LUTS.

Adult ; Humans ; Lower Urinary Tract Symptoms ; physiopathology ; therapy ; Male ; Middle Aged ; Prostatitis ; physiopathology ; therapy ; Urethra ; physiopathology ; Urinary Bladder Neck Obstruction ; physiopathology ; Urinary Bladder, Overactive ; physiopathology ; Urodynamics

Objective To investigate the role of autophagy and synaptophysin (SYP) in cognitive impairment in de?pressed rats receiving electroconvulsive shock (ECS). Methods Clean and healthy adult male Sprague-Dawley rats were acclimatized to a standard laboratory environment for 7 days. The chronic unpredictable mild stress (CUMS) was used to establish the rat model of depression. Behavior tests were conducted before and after CUMS to evaluate the depression and cognition level of rats. After establishment of the model, 24 rats were randomly divided into ESC group (group E) and depression group (group D) with 12 rats in each group. The rats in group E were administered 80 mg/kg of propofol (10 mg/mL) by intraperitoneal injection, followed by ECS treatment. The rats in group D were administered propofol by intra?peritoneal injection, followed by sham-ECS treatments. The above interventions were conducted daily for 7 consecutive days. After the interventions, rats underwent behavior tests as before. Subsequently, rats were killed and specimens were collected for measurements. Immunohistochemistry was performed to examine autophagy markers such as Beclin 1 and LC3Ⅱand ELISA was used to detect SYP in the hippocampus. Results Group E after ECS significantly increased the percentage of sucrose preference (68.2%±8.7%), rearing times (7.0±1.9), total horizontal distance [(569.5±70.0) cm], es? cape latency [(21.9±5.3)s] and space exploration time [(20.5±3.9)s] compared with group D or group E before ECS. There was no significant difference in these index between groups before ECS or in group E between before and after ECS(P>0.05). Compared with group D, group E had upregulated protein expression levels of Beclin 1 and LC3Ⅱin CA1, CA3, DG as well as the area near the hippocampus and increased SYP contents (P<0.05). Conclusions Cognitive impairment in depression rats following ECS correlates with activated autophagy and increased SYP by ECS.

Antibiotic-associated diarrhea(AAD)in children is a type of diarrhea that occurs after the use of antibiotics in children,and its pathogenesis is closely related to the intestinal flora.The medication of antibiotics can affect the metabolic function of the intestinal flora and the immune function of the body,and then leads to the occurrence of AAD.In the view of Chinese medicine,AAD in children is mainly involved the spleen,and the etiology of the disease is due to the weakness of the spleen and stomach of the body constitution together with the attack of the pestilential pathogen and the accumulation of drug toxin.The pathogenesis of ADD in children is characterized by spleen deficiency with predominant dampness,deficiency of spleen qi,and insufficiency of spleen yang.Spleen deficiency is the root cause of pediatric AAD,and spleen and intestinal flora have commonality,so the treatment of pediatric AAD can be performed from the perspective of the spleen.The treatment of pediatric ADD from the spleen follows the principle of strengthening and activating the spleen,and the regulation of the spleen for achieving the purpose of treating the disease from the root can be achieved by the methods of strengthening spleen and draining dampness,strengthening spleen and replenishing qi,and strengthening spleen and warming yang separately with the fundamental prescriptions of Shenlin Baizhu Powder,Sijunzi Decoction,and Fuzi Lizhong Pills.

Objective To study the biological characteristics of platelet-derived growth factor A and human beta defensin 2 (hPDGF-A/hBD2) gene-modified rat bone marrow mesenchymal stem cells (BMSCs). Methods By using liposome transfection technique, recombinant adenovirus vector expressing hPDGF-A/hBD2 (Adv-hPDGF-A-IRES-hBD2) labeled with GFP was transfected into 293T cells for virus packaging and amplification. BMSCs were isolated, cultured and infected by adenovirus-containing supernatant. The exogenous gene-modified BMSCs were comprehensively studied on their biological features, in terms of morphology, cell growth curve, cell cycle, and adipogenic, osteogenic and myogenic differentiation ability. Results hPDGF-A-IRES-hBD2 gene-modified BMSCs did not show obvious changes in cell viability, proliferation, cell cycle distribution or cell differentiation. Conclusion BMSCs were not only good carriers for exogenous hPDGF-A and hBD2 genes but also seed cells for cell therapy even after hPDGF-A/hBD2 modification.

Objective To explore the feasibility of mobilization circulating MSCs by G-CSF and observe the repairing effect of G-CSF mobilization in severe mouse traumatic brain injury(TBI) model.Methods MSCs-derived bone marrow and peripheral blood(PB) were cultured and its CFU-F were counted after mobilization by G-CSF.At 2,24,48,96,120,144,192,264,336 h after severe TBI in mice was establish,the neurobehavior of mice was measured by neurological examination and motor functional test,and mortality rate and pathologic changes were analyzed.Results MSCs-derived PB were successfully cultured.The CFU-F of mobilization group increased significantly than that of control group(P

Objective To evaluate the clinical effect of TAPAS intravascular closed catheter-directed thrombolysis combined with endovascular intervention treatment for Cockett syndrome complicated with deep venous thrombous of lower limbs. Methods The clinical data of 31 patients of Cockett syndrome complicated with deep venous thrombosis of lower limbs were retrospectively analyzed, and the patients were treated with TAPAS intravascular closed catheter-directed thrombolysis combined with endovascular intervention.Results Thirty patients were placed self-expandable stent,and 1 patient was treated by balloon dilation.The pain disappeared and the swelling decreased postoperatively.Twenty-two cases was cured (70.97%, 22/31), 7 cases showed excellence (22.58%, 7/31), and 2 cases showed effective (6.45%, 2/31), and there were no invalid patients. There was no pulmonary embolism and bleeding event during treatment. Conclusions The TAPAS intravascular closed catheter-directed thrombolysis combined with endovascular intervention treatment for Cockett syndrome complicated with deep venous thrombous of lower limbs is an effective and safe technique with satisfactory short term result.However,the long term clinical effect need to be further researched.

Objective To evaluate the cardio protective effect of astragaloside in rats with acute myocardial infarction and investigate the potential molecular mechanisms.Methods Forty-five Wistar rats were randomly divided into sham operation group,model group and astragaloside group,with 15 rats in each group.The acute myocardial infarction model was prepared by permanently occluding the left anterior descending artery in the model group and the astragaloside group,while the rats in the sham operation group were threaded only.Twenty-four hours after operation,the rats in the astragaloside group received astragaloside 20 mg · kg-1 · d-1 by intraperitoneal injection,once a day for two weeks.The rats in sham operation group and model group received isodose carboxymethylcellulose sodium by intraperitoneal injection,once a day for two weeks.Electrocardiogram was recorded to observe the changes of ST segment before ligation,after ligation immediately,1,3,7,14 d after intervened by astragaloside.Echocardiography was used to detect the left ventricular internal diameter at end-diastole (LVDD),left ventricular internal diameter at end-systole (LVDS),left ventricular end diastohc volume (LVEDV),left ventricular end systolic volume (LVESV),left ventricular fractional shortening (LVFS),left ventricular ejection fraction (LVEF).The levels of tumor necrosis factor alpha (TNF-α),interleukin (IL)-1 β,IL-6,nuclear factor-κB (NF-κB) in serum were measured by enzyme linked immunosorbent assay and the level of nitric oxide(NO) in plasma was measured by reagent kid.The expressions of toll-like receptor(TLR) 2,TLR4,nucleotide binding oligomerization do main(NOD) 1,NOD2,endothelial nitric oxide synthase (eNOS) mRNA were detected by quantitative real-time polymerase chain reaction.Results During the operation,there was 0,4,2 dead rats in the sham operation group,model group and astragaloside group.The ST segment in the model group and astragaloside group was higher than that in the sham operation group after operation immediately,1,3,7,14 d after intervented by astragaloside(P ＜ 0.01).The ST segment in the astragaloside group was lower than that in the model operation group at the time of 1,3,7,14 d after intervented by astragaloside(P ＜ 0.05).Compared with the sham operation group,the LVDD,LVDS,LVEDV,LVESV increased (P ＜ 0.01),LVFS,LVEF decreased (P ＜ 0.01),the levels of TNF-α,IL-1β,IL-6 and the expression of TLR2,TLR4,NOD1,NOD2 mRNA in cardiac muscle tissue were higher in the model group and astragaloside group (P ＜ 0.01),the level of NO was lower (P ＜ 0.05).The level of serum NF-κB in the model group was higher than that in the sham operation group(P ＜0.01),the expression of the eNOS mRNA in cardiac muscle tissue was lower than that in the sham operation group(P ＜ 0.05).There was no significant difference in the level of serum NF-κB and the expression of the eNOS mRNA in cardiac muscle tissue between the astragaloside group and sham operation group (P ＜ 0.05).Compared with the model group,in the astragaloside group the LVDD,LVDS,LVEDV,LVESV decreased (P ＜ 0.05),but LVFS,LVEF increased (P ＜ 0.05),the levels of TNF-o,IL-1 β,IL-6,NF-κB and the expression of TLR2,TLR4,NOD 1,NOD2 mRNA in cardiac muscle tissue were lower(P ＜ 0.05),the level of NO and the expression of the eNOS mRNA in cardiac muscle tissue were higher(P ＜ 0.05).Conclusion Astragaloside has protective effect against to acute myocardial infarction induced injury by decreasing the expression of TLR signal path and activating eNOS signaling.

Objective MR microscopy technique was used to study the visualization of senile plaque deposition in brains of the Alzheimer disease(AD)transgenic mice.Methods Two transgenic mice and 2 wild type mice at the age of 17 months were scanned in vivo using T_2 weighted image.After MR imaging,the brains were cut serially and immunostained according to the orthogonal pilot images.MR T_2 weighted images and immunohistological images of the senile plaque were observed and matched.Results The MR images showed that some black spots were visible in the hippocampus and cerebral cortex of the AD transgenic mice and some spots were consistent with the senile plaques on immunohistological sections.There were no spots in the MR images and the immunohistological sections of the wild type mice.Conclusion It is possible that MR microscopy can be used to detect the deposition of the senile plaque and diagnose AD specifically.

The chemical constituents were isolated and purified by various chromatographic techniques indluding silica gel, reverse phase silica gel, sephadex LH-20 and pre-HPLC and identified by their physicochemical properties and spectral data. Sixteen phenolic compounds had been isolated and n-butanol extracts which were fractionated from the ethanol extract of Oplopanax horridus roots bark. Their structures were identified as below, including 7 phenylpropanoid compounds, ferulic acid (1), 3-acetylcaffeic acid (2), caffeic acid (3), homovanillyl alcohol 4-O-beta-D-glucopyranoside (4), 3-hydroxyphenethyl alcohol 4-O-beta-D-glucopyranoside (5), 3, 5-dimethoxycinnamyl alcohol 4-O-beta-D-glucopyranoside (6), and 3-dimethoxycinnamyl alcohol 4-O-beta-D-glucopyranoside (7). Three coumarins, scopoletin (8), esculetin (9) and 3'-angeloyl-4'-acetyl-cis-knellactone (10). And 6 lignan compounds, (+)-isolaricires-inol-9'-O-beta-D-glucopyranoside (11), 3, 3'-dimethoxy-4, 9, 9'-trihydroxy-4', 7-epoxy-5', 8-lignan-4, 9-bis-O-beta-D-glucopyranoside (12), (+)-5, 5'-dimethoxylariciresinol 4'-O-beta-D-glucopyranoside (13), (-)-5,5'-dimethoxylariciresinol 4'-O-beta-D-glucopyranoside (14), (-)-pinoresinol 4'-O-beta-D-glucopyranoside (15), and (+)-5, 5'-dimethoxylariciresinol 9'-O-beta-D-glucopyranoside (16). All compounds were isolated and identified for the first time from this plant All the constituents except compounds 4, 6, 12 and 13 were obtained for the first time from the genus Oplopanax.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Oplopanax ; chemistry ; Phenols ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization