Objective To investigate the effect of mycophenolate mofetil (MMF) and sildenafil on the systolic pulmonary arterial pressure (SPAP),right ventricular hypertrophy index (RVHI),pulmonary arterial and heart structural of pulmonary arterial hypertension (PAH) rat models.Methods The rat models of monocrotaline (MCT)-PAH were developed.Sprague-Dawley male rats were randomly assigned into the control group,the MCT group,the MMF (40 mg·kg-1·d-1) group,the sildenafil (20 mg·kg-1·d-1) group,and the MMF (40 mg·kg-1·d-1) + sildenafil (20 mg·kg-1·d-1) group.The SPAP and RVHI were measured,and the pulmonary arterial and heart structural changes were observed for all rats.Statistical analysis were performed by one-way ANOVA and rank-sum test.Results ① SPAP of the MMF group,the sildenafil group and the MMF + sildenafil group were (31±8),(37±8),(29±6) mmHg,while that of the MCT group was (53±7) mmHg,the difference was statistically significant (P＜0.01).The RVHIs in the MMF group,the sildenafil group and the MMF+sildenafil group were reduced [(0.365±0.038),(0.407±0.047),(0.325 ±0.459) respectively] when compared with the MCT group (0.543±0.080),the difference was statistically significant (P＜0.01).The SPAP between the MMF+sildenafil group and the sildenafil group was statistically significantly different (P＜0.05),and the RVHI difference between the MMF+ sildenafil group and the sildenafil group was statistically significant (P＜0.05).② The wall thic-kness/tubes diameter of the MMF group (0.355±0.074) and the MMF+sildenafil group (0.289±0.017) were reduced when compared with that of the MCT group [(0.466±0.006)],the difference was statistically significant (P＜0.05).The wall thickness/tubes diameter of the MMF group (0.355±0.074) were reduced compared with the sildenafil group (0.455±0.006),and the difference was statistically significant (P＜O.05).In addition,the wall thickness/tubes diameter of the MMF+ sildenafil group (0.289±0.017) was reduced when compared with that of the sildenafil group (0.455±0.006),and the difference was statistically significant (P＜ 0.05).Conclusion Both sildenafil and MMF can reduce the SPAP and RVHI of PAH rat models induced by MCT.MMF and sildenafil can reduce wall thickness as well.

AIM: To investigate whether carnosine can inhibit cataract formation and protect Na+-K+ATPase against inactivation induced by a glucocorticoid.METHODS: Two hundred and twenty clear lenses cultured in vitro were randomly divided into five groups: control group (DMEM), steroid group (DMEM+Dexamethason 10μmol/L),lower concentration carnosine-treated group (DMEM+Dexamethason 10μ mol/L+Carnosine 2mmol/L), higher concentration carnosine-treated group (DMEM+Dexamethason 10μmol /L+Carnosine 5mmol/L) and carnosine group (DMEM + Carnosine 5mmol/L). Progression of cataract formation was evaluated daily using a dissecting microscope. On 1, 3, 5 and 7d, 10lenses of every group were homogenized and the activity of Na+-K+ATPase was measured by using spectrophotometer.RESULTS: During the incubation, mistlike opacity was observed in the lenses of the control group and carnosine group,but in the steroid group appeared dense nuclear opacity, while both two carnosine-treated groups came out visible demarcation between nuclear and cortical regions on 7d. A decrease in the activity of Na+-K+ATPase was found in the lens of the steroid group. On 3, 5, 7d, Na+-K+ATPase activity decreased 22.34% (P=0.002),47.98% (P＜0.001),75.37% (P＜0.001) compared with that at 1d, respectively. In the carnosine group,the activity of Na+-K+ATPase remained at the level of the control throughout the 7-d incubation, indicating that carnosine itself did not interfere with the original lens enzyme activity. In the lower concentration carnosine-treated group, on 3, 5, 7d,the activity of Na+-K+ATPase increased 10.8% (P＜0.05),44.6% (P＜0.01), 57.4% (P＜0.01) of control activity, respectively. In the higher concentration carnosine-treated group, on 3, 5, 7d, the activity of Na+-K+ATPase increased 11.3% (P＜0.05), 45.7% (P＜0.01), 57.6% (P＜0.01) of control activity,respectively. The activity of Na+-K+ATPase in both two carnosine-treated groups were only 6.7% and 6.5% lower than that of the control group after 7-d incubation. After the 7-d incubation, the Na+-K+ATPase activity of the lenses in the steroid group decreased significantly compared with carnosine-treated groups (P＜0.01).CONCLUSION: Carnosine prevents the cataract formation induced by a glucocorticoid, and significantly inhibits the inactivation of Na+-K+ATPase induced by a glucocorticoid.

Aspirin ; pharmacology ; Drug Resistance ; Humans

Background How to harvest the purified corneal endothelial seed cells is very important for the corneal tissue engineering technology. Herein,to establish a good culture method and effective identification method of corneal endothelial cells ( CECs) is the key. Objective Present study wag to establish the cultivating and identifying approach of the rabbit CECs and detect the biological characteristics of passaged cells. Methods Rabbits CECs were isolated from Descemet's membrane peeled off completely in 30 New Zealand white rabbits and then digested with 0. 25% trypsin-0. 02% EDTA and primarily cultured in CECs medium containing 15% fetal bovine serum. The growth situate and cellular morphology of rabbit CECs were observed under the inverted phase-contrast microscope following the alizarin red staining. Rabbit CECs were identified by cellular morphology as well as in gene and protein level, including the detection of expressions of collagen type IV α2(COL4A2) ,vascular endothelial growth factor receptor 2 ( FLK1 ) , Na+-K+ ATPase alpha 1subunit ( ATP1 A1 ) , aquaporin 1( AQP1 ) , voltage-dependent anion channels ( VDACs) by reverse transcription-polymerase chain reaction ( RT-PCR ). The expression and distribution of neurone specific enolase ( NSE) , Na+-K+ ATPase, zonula occludens-1 ( ZO-1 ) were also detected by immunocytochemistry under the fluorescence microscope. The proliferation activity of the passage cells was dynamically observed by MTT assay,and Na+-K+ ATPase activity of different generations cells was detected by ATPase kit. Results Majority of the primarily cultured cells adhered in 24 hours, infused in 2-3 days with the hexagon shape in appearance. The morphology of the cells was very varied with passage. Alizarin red staining showed a well- defined and well-lined cellular morphology similar to the corneal cells in vivo. Target genes of C0L4A2, FLK1, ATPIAI ,AQPI and VDACs were positively expressed in the cells. However,the expression of CK12 was absent in the cells. NSE, Na+-K+ ATPase, ZO-1 positive cells were respectively observed under the laser confocal scanning microscopy. MTT results showed a gradually low growth curve with the passage. Quantitative results also revealed that the Na+-K+ ATPase activity was gradually declined in different generations of cells ( F = 77. 174, P = 0. 000 ). Conclusion The enzyme digestion is a better approach of isolating and culturing comeal endothelial cells. Cultured cells can be identified by cells staining, gene expression and protein level. Earlier generation of CECs are ideal seed cells for cornea tissue engineering.

AIM: To study the effect of the subretinal fluid (SRF) on proliferation of retinal pigment epithelium (RPE) cells and retinal glial (RG) cells and associated activation and translocation of protein kinase C (PKC) as well as the application of PKC inhibitor.MTEHODS: RPE and RG cells were disintegrated to obtain PKC activity of cytoplasm and cellular membrane after being treated by the subretinal fluid (SRF) from the different stages of PVR patients (grade B and C) or being treated with PKC specific activator [phorbol-12-myris-tate-13-acetate (PMA)] or normal vitreous or DMEM culture medium. PKC activity in cytoplasm and cellular membrane was measured using radioactive isotope 32P labeling in a specific reaction of phosphorylation on PKC substrate. In addition, the PKC inhibitor, dequalinium chloride, was used to pretreat the RPE and RG cells before the cells exposed to SRF or PMA or normal vitreous. 3H-TdR (tritiated thymidine) was used to measure the levels of proliferation of RPE and RG cells with or without the activation and translocation.RESULTS: SRF and PMA promoted the proliferation of RPE and RG cells. SRF and PMA activated PKC in the cytoplasm of RPE and RG cells and the activated cytoplasm PKC translocated to the cellular membrane of RPE or RG cells. The cell proliferation or PKC activation or translocation were not equally active in RPE as in RG cells. However, PKC inhibitor which attenuated the cell proliferation did not show significant difference on inhibition of RPE and RG cell proliferation. (P ＞0.05).CONCLUSION: SRF can lead to the activation and translocation of PKC in RPE and RG cells, which promote the proliferation of RPE and RG cells. Dequalinium chloride can inhibit PKC activation and translocation hence slow down the cells proliferation.

Combined Modality Therapy ; Digestive System Surgical Procedures ; methods ; Gastrointestinal Neoplasms ; surgery ; Humans ; Treatment Outcome

Germany ; Hazardous Substances ; analysis ; Workplace

[Objective] To evaluate the therapeutic effect of Oleum Curcumae Aromaticae (OCA) on leukemia cell strain HL-60 and K562 by observing its sensitivity in inhibiting HL-60 and K562. [Methods] Subculture of HL-60 and K562 cells was performed on the 96-hole culture plate and OCA 100?L at the final concentration of 400, 200, 100 and 50mg/L respectively was added. The blank control was added with RPMI-1640 culture fluid. After 24, 48 and 72 hours of treatment, MTF 10?L (5mg/mL) was added and the inhibitory rate on the cells was assayed by detecting the absorption value at 570nm wavelength. [Results] The inhibitory effect of OCA on HL-60 and K562 increased with the treatment time. The difference of inhibitory rates 24, 48 and 72 hours after treatment were significant (P

AIM: To study the curative effect for patients with age related cataract and shallow anterior chamber after phacoemulsification.METHODS: Totally 38 patients (38 eyes) with age related cataract and shallow anterior chamber were selected and divided into two groups according to the depth of the anterior chamber, as mild shallow anterior chamber group (2-2.5mm) 23 eyes, high risk shallow anterior chamber group (<2.0mm) 15 eyes.Thirty-eight patients (38 eyes) with age related cataract with normal anterior chamber were as control group at the same period.All the patients received the operations by the same doctor and were followed up for 3mo.The observed items included visual acuity before and after operations, intraocular pressure, anterior chamber depth, corneal endothelial cell density and complications.RESULTS: There were no significant difference on visual acuity, intraocular pressure and corneal endothelial cell density between the two groups before operations (P>0.05).The visual acuity improved significantly after operation in both groups (P<0.05).Intraocular pressure after operation decreased significantly in both groups (P<0.05).Anterior chamber depth increased significantly after operation in both groups (P<0.05).Corneal endothelial cell density decreased significantly in both groups (P<0.05).There were no significant difference on anterior chamber depth, intraocular pressure and corneal endothelial cell density between the two groups at different time point after operations (P>0.05).Posterior capsular rupture occurred in shallow anterior chamber group in 1 eye, suspensory ligament rupture in 1 eye.Posterior capsular rupture and suspensory ligament rupture occurred none in normal anterior chamber group.Postoperative corneal edema occurred in 10 eyes (26%) in shallow anterior chamber group, which occurred in 3 eyes (8%) in normal anterior chamber group.The difference on the incidence was significant (P <0.05).CONCLUSION: Phacoemulsification should be taken timely for patients with age related cataract and shallow anterior chamber.The postoperative visual acuity can be improved and the anterior chamber depth can increase.The operation is safe and effective for those patients.

AlM: To evaluate the clinical efficacy of canalicular intubation combined with external dacryocystorhinostomy ( ext-DCR ) for treatment of chronic dacryocystitis and upper lacrimal duct stenosis.
METHODS:Thirty-three patients (33 eyes) with chronic dacryocystitis and upper lacrimal duct stenosis who underwent canalicular intubation combined ext-DCR were retrospective analyzed. The silicon tube was indwelt for 6mo. All cases were re-examined 1wk;1, 3, 6, 9mo post-operation, flow of tears, pus excretion and lacrimal duct clearance were observed.
RESULTS: Clear lacrimal duct ratio was 100% in all cases during 1wk~6mo post-operation; Silicon tube was removed 6mo post-operation, 3mo after tube removal, rechecking reported 32 cases with clear lacrimal duct (97%) and 1 case with resistant duct (3%). Two cases ( 6%) with minor lacerations, no other complications were observed.
CONCLUSlON: Canalicular intubation combined ext-DCR is an effective treatment for chronic dacryocystitis and upper lacrimal duct stenosis.