What drug dosage range is appropriate for treatment? What drug dosage range can maximally reduce the incidence of adverse drug reaction (ADR)? The gold zone method as a new method of evidence-based medical research was proposed to study those two blind areas of drug dosage in this article. Studying the dose-effect relationship, taking gold zone as the middle range and dividing empirical range into 3 sections were the key to study design. The evidence-based survey with extremely large sample showed a U-shaped rule existing between the antibiotics’ dosage and the incidence of ADR; and the dosage in gold zone appeared at the bottom of U-shaped curve. The gold zone method for determining dosage is a special breakthrough currently for solving those two blind areas of drug dosage
Objective To investigate the dose-dependent relationship of bone marrow mesenchymal stem cells(MSCs) transplantation in improving ischemic myocardial dysfunction? in a rat ischemic heart model. Methods Myocardial infarction was induced in 32 inbred F344 rats by acute ligation of the left anterior descending(LAD) coronary artery. One week after ligation, the ratswere randomized? into four equal groups, with eight rats in each group. Equal volume Iscove’s modified Dulbecco’s medium was injected in the control group, 1×103(group 1), 1×105(group 2), and 1×107(group 3) 5-bromodeoxyuridine (BrdU) labeled bone marrow MSCs were injected into the infarcted myocardium. Cardiac function was evaluated by ultrasound before the ligation of the LAD, before the transplantation and the 4th week after transplantation. The expressions of BrdU,Connexin43,Myosin heavy chain β(MHC), and smooth muscle actin α(α-SMA) were detected by immunofluorescence and immunohistochemistry at the 4th week after transplantation. The amount of functional vessels stained by α-SMA was counted simultaneously. Results At the 4th week? after transplantation, the ejection fraction(EF) in goup 2 was more significantly improved than that in group1(0.54±0.20 vs. 0.34±0.16, P＝0.004) and EF in group 3 was more significantly improved than that in group 2(0.71±0.24 vs. 0.54±0.20,P＝0.018), whereas no significant difference between group 1 and control group was detected (0.34±0.16 vs. 0.36±0.15,Pgt;0.05). The BrdU labeled MSCs could be found in host myocardium. The number of cells in group 2 by double staining both for BrdU and for MHC observed in ischemic myocardium were significantly more than that in group 1? (323.20±91.62 n/HP vs. 51.75±27.58 n/HP,P＝0.049) and the same was true between group 3 and group 2(409.75±106.65 n/HP vs. 323.20±91.62 n/HP,Plt;0.001), whereas the result of control group was negative.The majority of transplanted cells were found positive staining both for MHC and for Connexin43 in all groups. There were lots of positive staining of α-SMA whose form were partly irregular in ischemic myocardium indicating that there was neovascularization in group1 and control group. More neovascularization in group2 was found than that in group 1 (28.38±12.79 n/HP vs. 22.75±9.07 n/HP, P＝0015) and more neovascularization in group 3 was found? than that in group 2 (35.63±13.27 n/HP vs. 28.38±12.79 n/HP, P＝0.002) . Conclusion Transplanted into infarcted myocardium, bone marrow MSCs may have significant and dose-dependent potential for cardiomyogenesis with functional recovery from myocardial ischemia.
Objective To compare the analgesic effects of fentanyl, tramadol and flurbiprofen axetil during vitrectomy under local anesthesia. Methods One hundred and twenty patients who underwent vitrectomy were randomly divided into four groups, 30 patients in each group. Control group (Group C): normal saline were given; Fentanyl group （group F): fentanyl 1 mu;g/kg; Tramadol group （group T): tramadol 1 mg/kg; Flurbiprofen group （group K): flurbiprofen axetil 1 mg/kg. Mean arterial pressure (MAP), heart rate (HR), oxygen saturation (SpO2), sedation classification (OAA / S) and pain score (NRS) were recorded prior to drug administration (T0) and the beginning of surgery (T1), 5 min (T2), 15 min(T3), 30 min (T4) and the end of surgery (T5) . The incidence of analgesic remedy and adverse reactions were also recorded after surgery. Results In group F, MAP at T1 and T2 were significantly lower than T0 and that of the other three groups at the same time point (F=5.367,5.967;P＜0.05). MAP at each time point of the other three groups had no significant changes (P＞0.05). In Group C, HR decreased significantly at T3and T4compared to T0 (F=7.900, 6.767;P＜0.05). In Group F, HR decreased significantly at T2 compared to T0 (F=3.117，P＜0.05). HR at each time point of group T and group K had no significant changes (P＞0.05). In group F, SpO2at T1 was significantly lower than T0 and that of the other three groups at the same time point (F=7.352, P＜0.05). SpO2of group F, group T and group K had no significant changes within groups (P＞0.05). In Group F, the median of OAA / S classification at T1 were grade four, which were lower than that at T0 and that of the other three groups at the same time point (chi;2=12.935, P＜0.05). There was no significant changes of the median of OAA / S classification at each time point in the other three groups (P＞0.05). In group C, the median of NRS score was three at T1 and was two at T2 respectively, which were higher than that at T0 and that of Group F and group T at the same time point (chi;2=13.748,11.616; P＜0.05). There were no significant changes of the median of NRS score in group F, group T and group K within groups (P＞0.05). Analgesic remedy percentages in group C, group F, group T and group K were 16.7%, 3.3%, 3.3%, 6.7%, respectively. The incidence of adverse reactions in group C, group F, group T and group K were 30.0%、23.3%、3.3%、16.7%, respectively.Conclusion Tramadol had efficient analgesic effects and low rate of adverse reactions during vitrectomy under local anesthesia.
Objective To observe the effects of intravenous thrombolysis with urokinase for central retinal artery occlusion (CRAO). Methods A total of 115 CRAO patients diagnosed by fluorescence fundus angiography (FFA) were enrolled in this study. The patients included 61 males and 54 females, with a mean age of (56.7plusmn;15.2) years (from 41 to 75 years). The duration ranged from 1 to 30 days. All the patients were affected unilaterally. All the patients were received the treatment of intravenous thrombolysis with urokinase (3000 U/kg, two times per day, continuous treatment for six to seven days) and retrobulbar injection of dexamethasone 2.5 mg (one time per day, continuous treatment for 14 days). Following that, 1.2 mg/kg brain protein hydrolysate (nerve nutrition) and 360 mg troxerutin (vasodilator) were given by intravenous drip (one time per day, continuous treatment for 14 days). Effectiveness of the thrombolytic and subsequent treatments including the recovery of vision and retinal arterial filling time before and after treatment were observed. Comparing the visual acuity of post-treatment and pre-treatment, improving three lines or more is considered as effective markedly, improving two lines as effective, no change or a decline as no effect. With FFA as the retinal circulation recovery index, the arm-retinal circulation time (A-Rct ) le; 15s and all branches of central retinal artery were filled with fluorescence within 2s filling (normal) as effective markedly; A-Rct improved but was in 15 - 20s range, all branches of central retinal artery were filled with fluorescence within 3～8s as effective; A-Rct improved but was still ge; 21s, all branches of central retinal artery were filled with fluorescence within ge;9s as no effect. The relationship between age, gender, the disease course, subsequent treat time and curative effectiveness were analyzed. Results There were 79 patients were examined for FFA again after thrombolysis treatment which including 11 patients with complete obstruction and 68 patients with incomplete obstruction. In 11 patients with complete obstruction, eight patients showed that optic disc vascular retrograde filling disappeared, A-Rct was 28-54s, and the filling time from retinal artery to tip was 18 - 55s; three patients showed persistent optic disc vascular retrograde filling within 3 - 4 minutes of FFA. In 68 patients with incomplete obstruction, A-Rct returned to normal in 35 patients (51.4%), effective in 18 patients (26.5%) and no effect in 15 patients (22.1%). Retinal circulation time was shorter than that before thrombolysis treatment (chi;2=11.4, Plt;0.05). Comparison of distribution of visual acuity before and after thrombolysis treatment, the difference was statistically significant (chi;2=12.1, Plt;0.05). Comparison of distribution of final visual acuity after subsequent treatment with that of after thrombolysis treatment, 48 eyes improved two lines or more, the efficiency was 41.7%, the difference was statistically significant (chi;2=14.6, Plt;0.05). Comparison to that of before treatment, vision changes showed effect markedly in 58 patients (50.4%), effective in 35 patients (30.4%), no effect in 22 patients (19.2%), the difference was statistically significant (chi;2=44.5, Plt;0.05). Comparison the average age to that of effective, valid and invalid patients, the difference was not statistically significant (t=0.98, 1.17, 0.55; Pgt;0.05). There was no relationship between effectiveness and gender (chi;2=2.6, Pgt;0.05). In 76 patients with duration within seven days, 43 patients were effective markedly and 22 patients were effective, the efficiency was 85.5%. In 25 patients with duration of 8 - 15 days, 11 patients were effective markedly and eight patients were effective, the efficiency was 76.0%. In 34 patients who received subsequent treatment 8 - 14 days, 18 patients were effective markedly and nine patients were effective, the efficiency was 79.4%. In 51 patients who received subsequent treatment 15-21 days, 27 patients were effective markedly and 18 patients were effective, the efficiency was 88.2%. Conclusion Intravenous thrombolysis with urokinase was effective in the treatment of CRAO.
Objective To observe the influence of cisplan on the expression of B7-H1 in retinoblastoma (RB) cells，and to investigate its mechanism. Methods Human RB cell line HXO-Rb44 cells were treated by 6 different concentrations of cisplan (0.000, 0.375, 0.750, 1.500, 3.000, 6.000 mu;g/ml), and their B7-H1 mRNA expression was determined by the reversetranscription polymerase chain reaction (RT-PCR) and fluorescence quantitative PCR (FQ-PCR); the B7-H1 protein expression was determined by immunofluorescence and flow cytometry. HXO-Rb44 cells were treated by 1.5 mu;g/ml cisplan for 0, 15, 30, 60, 120 min, then the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) was detected by Western blot.Results The expression of B7-H1 mRNA and protein in the 0.375, 0.750, 1.500, 3.000, 6.000 mu;g/ml group were significantly higher than that of the blank control group (F=395.478，112.03; P=0.000）. Western blot showed that cisplan (1.5 mu;g/ml) could activate ERK1/2 by increasing its phosphorylation in HXO-Rb44 cells. After cisplan treatment, the phosphorylation of ERK1/2 increased gradually and reached its peak at 30 min, and then went down gradually.Conclusion Cisplan can promote the expression of B7-H1 and activate ERK1/2 in RB cells.
Objective To observe the influence of the indomethacin on the proliferative and invasive activity of OCM-1 human choroidal melanoma cells. Methods OCM-1 cells were cultured with different concentrations of indomethacin (25, 50, 100, 200, 400 mu;mol/L ), and their proliferation were assessed by methyl thiazolyl tetrazolium（MTT）, invasive behaviors were examined by cell invasion assays, expression of survivin and VEGF were evaluated by reverse transcriptase polymerase chain reaction（RT-PCR）, immunofluorescence staining, ELISA and western blot analysis. Result All concentrations of indomethacin in this study can inhibit the proliferation and invasion of OCM-1 cells in a time and dosage-dependant manner（MTT/24 h：F=19.642，P＜0.01；MTT/48 h：F=136.597，P＜0.01；MTT/72 h：F=582.543，P＜0.01；invasion assays：F=54.225，P＜0.01）. Immunofluorescence staining indicated that survivin and VEGF mainly expressed in the cytoplasm of OCM-1 cells. Survivin mRNA in OCM-1 cells was inhibited by 100, 200, 400 mu;mol/L indomethacin（F=16.679，P＜0.01）. The concentrations of survivin were (787.3plusmn;47.37), (257.0plusmn;26.21), (123.3plusmn;8.02) pg/ml in control group and 100, 400 mu;mol/L indomethacin groups, respectively. Survivin expression was also significantly down-regulated in indomethacin-treated cells by Western blot analysis.Indomethacin had no effects on VEGF expression in OCM-1 cells.Conclusions Indomethacin can inhibit proliferation and invasion of OCM-1 cells in vitro,down-regulated expression of survivin may be the mechanism.
Objective To observe the inhibition effect of selective cyclooxygenase2 inhibitor（celecoxib）on the experimental choroidal neovascularization（CNV）. Methods Thirty 8-10 weeks old healthy male Brown-Norway（BN）rats were randomly divided into the control, laser and celecoxib group,with 10 rats in each group. At the dosage of 50 mg/kg, celecoxib was gavaged twice per day. After 7 days, experimental CNV was induced by Krypon laser on laser group and celecoxib group. Fundus fluorescein angiography (FFA) was performed on days 3, 7,14,21,30 after laser photocoagulation.On days 21 after photocoagulation, 5 rats in each group were sacrificed and the relative thickness of CNV membranes, the expression of COX-2, vascular endothelial growth factor(VEGF) and matrix metalloproteinase-2(MMP-2) were studied by histopathologic or immunohistochemistry examination.Results On days 21 after photocoagulation, the incidence of CNV in the celecoxib group is significantly lower than that in the laser group (chi;2=7.1068,P=0.0077); the relative thickness of the CNV membranes in the celecoxib group is reduced 41.38% compared to the laser group, the difference is statistically significant (t=16.760 0，P=0.0000).COX-2,VEGF and MMP-2 expression in the CNV membrane of celecoxib group were significantly lower than in control group (t=5.710 0,5.840 0, 8.020 0; P=0.000 0); the COX-2, VEGF and MMP-2 expressions in choroid and retina of control group were weak. Conclusion Prophylactic celecoxib can reduce the expression of VEGF and MMP-2 by inhibiting COX-2, and prevent the CNV induced by laser photocoagulation.
Objective To evaluate the safety repeated intravitreal injection of bevacizumab (Avastin) with different dosage in rabbitsprime;eyes. Methods Fourteen chinchilla rabbits were randomly divided into 3 groups, including both eyes of 2 rabbits in the control group,the right eyes of the other 12 rabbits in the experimental group,and the left eyes of the 12 rabbits in the experimental control group. The eyes in the experimental group underwent intravitreal injection of bevacizumab with the dosage of 2.5 mg/0.1 ml and 5.0 mg/0.2 ml of bevacizumab; the eyes in the experimental control group underwent intravitreal injection of normal saline with the same dosage as in the experimental group. Injections were performed every two weeks and lasted six weeks. Clinical observation and retinal function tests were performed before and two days after every injection. The eyes were sacrificed 1 week and 4 weeks after last intravitreal injection respectively.Electron and optical microscope and TUNEL were performed.Results After intravitreal injection,no obvious anterior chamber flare, abnormal change of the ocular fundus, or vitreous opacity and hemorrhage was observed in all of the eyes.No change was found by indirect ophthalmoscope,Bultrasonic inspection, ultrasound biomicroscopy and optical coherence tomography. The number of anterior chamber flare before and after the injection with the dosage of 2.5 and 5.0 mg, the difference among the 3 groups didnprime;t differ much from each other (Pgt;0.05).Amplitude and pattern of ERG responses and flash VEP were similar between the control and experimental groups (Pgt;0.05). Some inflammatory cells were found in the some bevacizumabinjected eyes 1 week after injection, and vanished 3 weeks later. The histological configuration of the retina didnprime;t change in both experimental control and the control group. Electron microscopy showed that plasma cells were presented and vacuolelike change was observed in part of the photoreceptor cells in 5.0 mg experimental group 1 week after injections.Cellular apoptosis was observed in the photoreceptor cell layer. The number of apoptotic cells was more in 5.0 mg experimental group than that in the control and experimental control group 1 week after injections (Plt;0.01). Conclusion Multiintravitreal injection with 5.0 mg bevacizumab may have mild toxicity to the retina in the rabbits.
Objective To observe the inhibition effect of curcumin on the proliferation of rabbit retinal pigment epithelial (RPE) cells and investigate its mechanism. Methods The 4th generation of RPE cells were selected and divided into curcumin group and blank control group. The concentration of curcumin included 10, 15, and 20 mu;g/ml. The MTT assay was used to evaluate the inhibition effect on the proliferation of RPE cells at the 24th, 48th, 72nd and 96th hour after cultured with curcumin (10, 15, and 20 mu;g/ml). The IC50 value of curcumin at different time points were calculated by Linear Regression. Flow cytometry was used to detect the effect on the cell cycle at the 72nd hour after cultured with curcumin (15 mu;g/ml); the expression and apoptosis of proliferating cell nuclear antigen (PCNA) were also determined at the 24th,48th, and 72nd hour after cultured with curcumin (15 mu;g/ml) respectively. The configuration of RPE cells were observed by transmission electron microscope. Results The IC50 value of curcumin at the 24th,48th, 72nd and 96th hour was 29.31, 17.50, 13.24, and 10.99 mu;g/ml respectively. Cell cycel analysis indicated that curcumin blocked cells in G0/G1 phase. At the 24th, 48th, and 72nd hour after cultured with curcumin (15 mu;g/ml), the expression of PCNA of RPE cells were 565.04plusmn;23.60, 473.61plusmn;36.88, and 396.15plusmn;32.45; the apoptosisrate were （12.83plusmn;0.13）%，（32.27plusmn;4.51）%，（56.81plusmn;8.67）%, respectively. The differeces of curcumin groups compared with the control group were significant (P＜0.05). Apoptosis of RPE cells was observed under transmission electron microscope. Conclusions Curcumin can inhibite the proliferation of RPE cells by inhibit the synthesization of PCNA and inducing the apoptosis of RPE cells. Curcumin may become a potential drug to prevent and treat PVR.
Objective To observe the effect of medicineinduced posterior vitreous detachment (PVD) on proliferative vitreoretinopathy (PVR). Methods PVR was induced in the left eyes of 24 pigmented rabbits by intravitreal injection with platelet rich plasma. The rabbits were randomly divided into two experimental groups (group A and B) and one control group with 8 eyes in each group. Three hours later, the eyes in group A and B and the control group underwent intravireal injection with 1 U plasmin 0.05 ml+20 U hyaluronidase 0.05 ml, plasmin 0.1 ml, and balance salt solution 0.1 ml, respectively. The grade of PVR was recorded 1, 7, and 28 days after the intravitreal injection, and the eyes were examined by flash electroretinogram (FERG), B-scan, and retinal histopathological examination. Results The PVR models of rabbit eyes were induced successfully. On the 7th day after injection, complete and partial PVD was found in 5 and 3 eyes respectively in group A; partial PVD in 5 eyes and no complete PVD was observed in group B; there was no PVD in the other 3 eyes in group B and also in the eyes in the control group. On the 28th day after intravitreal injection, PVR grade of group A and B were both obviously lower than that of the control group（D=75.6, 98.9；P=0.003，P=0.011）; On the 7th and 28th day after injection, the b-wave amplitude in group A and B was significantly higher than that in the control group; PVR grade of the PVD eyes was lower than that of nonPVD eyes; PVR grade of the complete PVD eyes was only 0～1. Conclusions Three hours after the PVR models of rabbit eyes were induced, complete PVD induced by intravitreal injection of plasmin combined with hyaluronidase could prevent the development of PVR of rabbit eyes in some degree; partial PVD induced by plasmin alone or combined with hyaluronidase could relieve the development of PVR.