Background Anti-VEGF drugs are generally applied in the treatment of ocular neovascular diseases.However,the therapy effect is unsatisfactory in some patients.Studing the effect of hypoxia-inducible factor-1 (HIF-1),a upstream regulatory gene of VEGF,and its limiting enzyme prolyl-4-hydroxylase domain proteins (PHDs) is of important clinical significance.Objective This study was to investigate the negtive regulation of exogeneous PHDs on HIF-1 pathway in human RPE cells.Methods pFLAG-PHD1,pFLAG-PHD2 and pFLAG-PHD3 plasmids were constructed by extracting RNA from Hela cell line and coloning PHD1,PHD2 and PHD3 using reverse transcription PCR with restriction enzyme.The plasmids were identified by gene sequencing.ARPE-19 cells were cultured at 21％ O2 (normoxia group),1％ O2 (hypoxia group),or in hypoxia-mimicking agents (CoCl2,anoxia group),respectively,and then were transfected with plasmids encoding FLAG-tagged PHD1,PHD2,PHD3 and pFLAGCMV2 transfected cells served as blank control.The expressional intensities of PHD1,PHD2 and PHD3 in the cells were detected and compared among different groups by using Western blot assay.The transcriptional activity of HIF-1 in the cells was evaluated with dual luciierase reporter assay.Results Western blot assay showed that PHD1,PHD2 and PHD3 all were expressed in ARPE-19 cells in the normoxia group,hypoxia group and anoxia group.The expression was strong in PHD2 protein and was weak in PHD3 protein,a statistically significant difference was found between PHD2 protein expression and PHD1 or PHD3 expressions (all at P＜0.05).Endogenous HIF-1 activity was elevated in pFLAG-CMX transfected cells in the hypoxia group and anoxia group than that in the normoxia group.Compared with pFLAG-CMX transfected cells,no obvious change was seen in the endogenous HIF-1 activity in the normoxia group,however,HIF-1 activity was declined in the hypoxia group and anoxia group after pFLAG-PHD1,pFLAG-PHD2 or pFLAG-PHD3 transfection.Under the same oxygen environment,HIF-1 activity was lower in the pFLAG-PHD2 transfected cells than that in the pFLAG-PHD1 or pFLAG-PHD3 transfected cells (both at P＜0.05).Conclusions PHDs play a negative regulation to HIF-1 activating pathway in human RPE cells,especially in hypoxia and anoxia cells.Among PHDs proteins,PHD2 presents the strongest inhibition on HIF-1 activating pathway.

OBJECTIVE: To evaluate the effect of Xiangdan Injection on mRNA expression of the endothelial vaso-active factors of patients with coronary heart disease and blood stasis. METHODS: Fifty-six patients were randomly divided into two groups:twenty-eight patients were treated according to the therapeutic guide for coronary heart disease as the control group and 28 were given the same treatment plus Xiangdan Injection as the treated group. The expressions of ET-1 and eNOS mRNA were examined with RT-PCR before experiment and ten days later. RESULTS: The positive rate of eNOS mRNA of the treated group increased, while the positive rate of ET-1 mRNA of the treated group decreased after ten day's treatment, with significant differences as compared with that before the experiment. Xiangdan Injection up-regulated the eNOS mRNA expression and suppressed the ET-1 mRNA expression. Changes of expression were not observed in the control group. CONCLUSION: Xiangdan Injection improves the endothelial function of patients with coronary heart disease and blood stasis by regulating the expressions of ET-1 and eNOS mRNA.

Objective To observe the effects of different traditional Chinese medicine therapeutic methods on heart function and red cell volume distribution width (RDW) in patients with chronic heart failure (CHF). Methods A randomized, blinded and controlled study was conducted. One hundred and ninety patients with CHF accompanied by yang deficiency syndrome in Department of Cardiology of Chengdu Municipal First People's Hospital were divided into five contrast groups according to randomized envelope method:namely control group, warming yang group, nourishing yin group, activating blood group and combined warming yang and nourishing yin therapeutic group (combined group), each group being 38 cases. The patients not consistent with the criteria to enroll into the study and those lost contact were excluded from the study, thus the final total patients were 183 in number, including control 37, warming yang 36, nourishing yin 36, activating blood 37 and combined group 37 cases. All the cases were treated with routine medical therapy in accord to the guideline for CHF. In addition, placebo oral liquor was given to the control group, Wenyangjianxinling oral liquor (main ingredients:Aconiti Lateralis Radix Preparata, Astragali Radix, Chuanxiong Rhizoma, Epimedii Folium, Periplocae Cortex, Descurainiae Semen, Ginseng Radix et Rhizoma Rubra) was given to the warming yang group, Ophiopogonis Radix and Anemarrhenae Rhizoma oral liquor (main ingredients:Ophiopogonis Radix,Anemarrhenae Rhizoma) was taken by the nourishing yin group, Salviae Miltiorrhizae Radix and Chuanxiong Rhizoma oral liquor (main ingredients: Salviae Miltiorrhizae Radix, Chuanxiong Rhizoma) was administered by the activating blood group, and the main ingredients of oral liquors given to the warming yang and nourishing yin groups were assigned to the combined group, each group 10 mL, three times a day for one year. According to the prognosis, the patients were divided into survival group (168 cases) and death group (15 cases). The re-hospitalization situations such as aggravation of heart failure, acute myocardial infarction, angina, stroke, etc. were observed. The changes of hemoglobin (Hb) and RDW in the survival and death groups were investigated and compared between them. Results Compared with the control group, the rate of re-hospitalization was significantly decreased in the activating blood group [27.03% (10/37) vs. 54.05%(20/37), P<0.05];there were no statistical significant differences among all the other groups (all P>0.05). Before treatment, the levels of left ventricular ejection fraction (LVEF), fractional shortening (FS), the ratio of early to late ventricular filling velocities (E/A ratio) and E deceleration time (DT) showed no significant differences among five groups (all P>0.05). Compared with the control group at the same period, the LVEF, FS, E/A and DT were significantly increased in activating blood group and combined group after the treatment [LVEF: 0.453±0.131, 0.448±0.104 vs. 0.394±0.112, FS:(27.9±9.8)%, (27.0±11.5)%vs. (22.2±13.3)%, E/A:0.88±0.16, 0.92±0.20 vs. 0.75±0.27, DT (ms): 265.4±30.3, 251.4±37.5 vs. 225.7±35.4, all P < 0.05]. FS and DT were significantly increased in warming yang group after treatment [FS: (26.4±10.3)% vs. (19.7±7.4)%, DT: 242.0±38.7 vs. 216.3±50.9, both P < 0.05]. In the activating blood group, the level of RDW was elevated after treatment compared with that before treatment, but in the comparison with that in the control and nourishing yin groups, it was still obviously lower [(12.98±2.97)%vs. (14.37±2.52)%, (13.05±2.36)%, both P < 0.05]. The comparisons of Hb and RDW among other groups had no statistical significant differences (all P > 0.05). The level of Hb in death and survival groups also had no statistical significant difference (P > 0.05). The RDW of dead group was markedly higher than that of the survival group [(14.39±2.17)%vs. (13.02±2.08)%, P<0.05]. Conclusion The level of RDW is related to the prognosis of CHF patients, the level of RDW in death group is higher than that in survival group, and the activating blood circulation and removing blood stasis therapy can decrease the level of RDW, reduce the rate of re-hospitalization and improve the heart function in CHF patients.

Abstract Myopia is a common refractive error in the process of development. With the high incidence and low age of myopia in children and adolescents, it has become a worldwide public health problem, seriously endangering physical and mental health. The prevention and control of myopia has a long way to go. Low concentration atropine, as an M type choline receptor inhibitor, has been proved by medical research to effectively delay the progress of myopia. This paper reviews the relevant studies at home and abroad in recent years. The optimal concentration of low concentration atropine, the best clinical regimen and possible potential side effects were discussed. From the point of view of school public health, this paper discusses the significance of low concentration atropine for the prevention and control of myopia among school age students, in order to provide a reliable basis and new ideas for the follow up clinical application of atropine and myopia prevention and control strategies for school students.

OBJECTIVETo prepare the solid dispersion of tanshinone II A (TS II A) by the combined application of nano-silica and poloxamer 188 (F68), in order to observe its dissolution and stability.

METHODTanshinone II A solid dispersion was prepared by the solvent method with nano-silica and poloxamer 188 as binary vectors. Its physical characteristics, in vitro dissolution and stability were further assessed.

RESULTThe tanshinone II solid dispersion was prepared with the weight ratio of nano-silica and poloxamer 188 of 1: 3. The differential scanning calorimetry (DSC) demonstrated that Tanshinone II A existed in vectors as amorphous state. The in vitro dissolution of tanshinone II A solid dispersion is up to 90% at 60 min. Accelerating experiment showed that content and in vitro dissolution of tanshinone II A solid dispersion did not change after storage over 3 months.

CONCLUSIONSolid dispersion of binary vector of tanshinone II A can obviously improve the dissolution and stability of tanshinone II in practice.

Calorimetry, Differential Scanning ; Diterpenes, Abietane ; chemistry ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Solubility

Objective:To investigate the effects of ginkgolide B on neurological function recovery and the Wnt/β-catenin pathway after ischemic stroke in mice.Methods:Fifty-five C57/BL6 mice were selected, of which 10 mice were kept as the sham group and the remaining 45 mice were constructed as the ischemic stroke model. There were 40 mice who finally completed the modeling, and then they were randomly divided into the blank control group (GB0w), short-course administration group (GB1w), long-term administration group (GB2w), and long-term administration+antagonist group (GB2w+PRI-724), with 10 mice in each group. There was no drug intervention after MCAO in GB0w. The mice in GB1w were given ginkgolide B (10 mg/kg) 0.1 ml within 1 week after MCAO; in GB2w were given ginkgolide B (10 mg/kg) 0.1 ml within 2 weeks after MCAO; and in GB2w+PRI-724 were nasally fed ginkgolide B (10 mg/kg) 0.1 ml within 2 weeks after MCAO; and selective antagonist PRI-724 was given 3 h before administration of ginkgolide B on days 8 to 14. Neurological function scores, walking on rotor bar test scores, expression of transforming growth factor-β1 (TGF-β1), fibroblast growth factor 4 (FGF4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), Wnt, β-catenin, and glycogen synthase kinase-3β (GSK-3β) were compared among the groups.Results:Compared with the sham group, the expressions of MDA, TNF-α, IL-6, FGF4, and GSK-3β in GB0w, GB1w, GB2w, and GB2w+ PRI-724 were increased, and the expressions of GSH-Px, SOD, TGF-β1, β-catenin, and Wnt were decreased (all P < 0.001). Compared with GB0w, the expressions of SOD, GSH-Px, TGF-β1, Wnt, and β-catenin were increased in GB1w, GB2w, and GB2w+PRI-724, and the expressions of MDA, TNF-α, IL-6, FGF4, and GSK-3β were decreased (all P < 0.001). Compared with GB1w, the expressions of GSH-Px, SOD, TGF-β 1, Wnt, and β-catenin were increased in GB2w and GB2w+PRI-724, and the expressions of IL-6, TNF-α, MDA, FGF4, and GSK-3β were decreased (all P < 0.001). Compared with GB2w, the neural function score, walking on the stick test score, and expressions of IL-6, TNF-α, FGF4, MDA, and GSK-3β were increased in GB2w+PRI-724, while the expressions of GSH-Px, TGF-β1, SOD, Wnt, and β-catenin were decreased (all P < 0.001). Conclusions:Ginkgolide B can effectively improve the neurological function of ischemic stroke mice and may be related to the Wnt/β-catenin pathway.

ObjectiveTo investigate the regulatory effects of Ginkgolide B on the biological characteristics of brain T cells and their interactions with glial cells during the recovery phase of ischemic stroke in mice. Methods36 adult C57BL/6 mice were randomly assigned to three groups: sham-operated group (Sham group), control group (PBS group), and Ginkgolide B treatment group (GB group). The Sham group underwent only sham surgeries, whereas the PBS and GB groups were subjected to a middle cerebral artery occlusion (MCAO) model using the filament method, followed by intranasal administration of an equivalent volume of either PBS or Ginkgolide B solution for 14 days post-injury. Neurological function changes were evaluated in all three groups using the rotarod test and a neurological scoring system. On day 15, single-cell sequencing was performed on fresh tissues from the brain injury areas, surrounding cortex, corpus callosum, and striatum of mice in the PBS and GB group to assess the biological characteristics of T cells and their subpopulations, and further explore the interactions and mechanisms among T cells, microglia, and oligodendrocytes. ResultsCompared with the Sham group, both PBS and GB group exhibited significant improvements in neurological scores and reduced pre-fall motor durations (P < 0.001). Compared with the PBS group, the GB group showed a downward trend in neurological scores and an upward trend in pre-fall motor durations on days 5, 10, and 15 post-ischemic brain injury, with a significant increase in pre-fall motor duration on day 15 (P < 0.05). Compared with the PBS group, the GB group exhibited a significant increase in T cell proliferative activity in the brain 15 days post brain injury (P < 0.05). The number of proliferative T cells and the levels of lipid metabolism were significantly elevated (P < 0.05), and there was a significant increase in extracellular matrix remodeling in all T cells (P < 0.05). Additionally, the interactions between T cells and both microglia and oligodendrocytes, as well as among the microglia themselves and between microglia and oligodendrocytes, were significantly enhanced in the GB group. This was primarily evident in the strengthened interactions between CD74 and macrophage migration inhibitory factor (MIF), as well as colony stimulating factor 1 receptor (CSF1R) and colony stimulating factor 1 (CSF1) (P < 0.05). However, the inflammatory levels of T cells showed no significant differences compared with the PBS group. ConclusionA mouse model of ischemic stroke can be successfully established by MCAO operation. Ginkgolide B may promote neurological recovery post-brain injury in mice by modulating the biological characteristics of T cells within the brain and their interactions with glial cells.