Mechanism of Liuwei Dihuang Pills in treatment of mice with diminished ovarian reserve based on proteomics / 中国中药杂志

This study aims to investigate the efficacy and possible mechanism of Liuwei Dihuang Pills in the treatment of diminished ovarian reserve(DOR) by using proteomic techniques. Firstly, cyclophosphamide(60 mg·kg~(-1)) combined with busulfan(6 mg·kg~(-1)) was injected intraperitoneally to establish the mouse model of DOR. After drug injection, the mice were continuously observed and the success of modeling was evaluated by the disturbance of the estrous cycle. After successful modeling, the mice were administrated with the suspension of Liuwei Dihuang Pills by gavage for 28 days. At the end of the gavage, four female mice were selected and caged together with males at a ratio of 2∶1 for the determination of the pregnancy rate. Blood and ovary samples were collected from the remaining mice on the next day after the end of gavage. Hematoxylin-eosin(HE) staining and transmission electron microscopy(TEM) were then employed to observe the morphological and ultrastructural changes in the ovaries. The serum levels of hormones and oxidation indicators were measured by enzyme-linked immunosorbent assay. Quantitative proteomics techniques were used to compare the ovarian protein expression before and after modeling and before and after the intervention with Liuwei Dihuang Pills. The results showed that Liuwei Dihuang Pills regulated the estrous cycle of DOR mice, elevated the serum levels of hormones and anti-oxidation indicators, promoted follicle development, protected the mitochondrial morphology of ovarian granulosa cells, and increased the litter size and survival of DOR mice. Furthermore, Liuwei Dihuang Pills negatively regulated the expression of 12 differentially expressed proteins associated with DOR, which were mainly involved in lipid catabolism, inflammatory response, immune regulation, and coenzyme biosynthesis. These differentially expressed proteins were significantly enriched in sphingolipid metabolism, arachidonic acid metabolism, ribosomes, ferroptosis, and cGMP-PKG signaling pathway. In summary, the occurrence of DOR and the treatment of DOR with Liuwei Dihuang Pills are associated with multiple biological pathways, mainly including oxidative stress response, inflammatory response, and immune regulation. "Mitochondria-oxidative stress-apoptosis" is the key to the treatment of DOR by Liuwei Dihuang Pills. YY1 and CYP4F3 may be the key upstream targets that trigger mitochondrial dysfunction and ROS accumulation, and the metabolism of arachidonic acid is the main signaling pathway of drug action.

Vitreous properties in patients with diabetes mellitus and its relationship with proliferative diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

When it comes to diabetic patients, persistent hyperglycemia and associated pathological conditions will not only cause diabetic retinopathy(DR)but also have an impact on the metabolism of vitreous, leading to diabetic vitreopathy. Owing to the adjacent anatomical position between the vitreous and retina, diabetic vitreopathy and DR are mutually promoted. Changes in vitreoretinal interface such as posterior vitreous detachment(PVD)and vitreoschisis, provide a scaffold for fibrovascular proliferative membrane and are closely associated with pars plana vitrectomy(PPV). This article sorts out the variation of diabetic patients' vitreous structure and biochemical components, along with the changes in the vitreous-retinal interface, particularly for the related research on its relationship with proliferative diabetic retinopathy(PDR), aiming at providing further cognition of diabetic vitreopathy as well as references for DR treatment and formulation of PPV.

Specific antitumor activity and mechanism of protonic bis-phenanthroline / 药学学报

Hepatocellular carcinoma (HCC) is a common malignant tumor worldwise. The incidence rate of HCC is high and is easy to metastasis and recurrence, which seriously affects human health. Traditional chemical drugs have some challenges such as toxicity, side effects, and multidrug resistance, thus it is urgent to find new drugs and effective targets. Here we synthesized a novel chemical, protonic bis-phenanthroline (H-BP), and the antitumor effect was investigated in the study. The results showed that H-BP could selectively inhibit the proliferation of tumor cells and cause HCC apoptosis. And also, in HCC tumor-bearing mice, H-BP could effectively prevent the growth of tumor mass, even completely eliminate the tumor at medium dose (5 mg·kg-1) and high dose (10 mg·kg-1), and meanwhile H-BP has no significant effect on the body weight of mice. The experimental protocol was approved by the Animal Ethics Committee of Southwest University, and the experimental operation was strictly carried out in accordance with the ethical principles of animal use and care. Mechanism studies showed that H-BP induced HCC apoptosis was related to down-regulation the expression of pleomorphic adenoma gene like-2 (PLAGL2), a oncogene transcription factor, resulting in the down-regulation of PLAGL2 downstream proteins hypoxia inducible factor and β-catenin. This study not only introduces the dimerization method to form novel compounds that will provide a new approach for drug design, but also suggests that PLAGL2 may be an effective target in tumor therapy.

The bioinformatics and metabolomics research on anti-hypoxic molecular mechanisms of Salidroside via regulating the PTEN mediated PI3K/Akt/NF-κB signaling pathway / 中国天然药物

Salidroside (SAL), a major bioactive compound of Rhodiola crenulata, has significant anti-hypoxia effect, however, its underlying molecular mechanism has not been elucidated. In order to explore the protective mechanism of SAL, the lactate dehydrogenase (LDH), reactive oxygen species (ROS), superoxide dismutase (SOD) and hypoxia-induced factor 1α (HIF-1α) were measured to establish the PC12 cell hypoxic model. Cell staining and cell viability analyses were performed to evaluate the protective effects of SAL. The metabolomics and bioinformatics methods were used to explore the protective effects of salidroside under hypoxia condition. The metabolite-protein interaction networks were further established and the protein expression level was examined by Western blotting. The results showed that 59 endogenous metabolites changed and the expression of the hub proteins of CK2, p-PTEN/PTEN, PI3K, p-Akt/Akt, NF-κB p65 and Bcl-2 were increased, suggesting that SAL could increase the expression of CK2, which induced the phosphorylation and inactivation of PTEN, reduced the inhibitory effect on PI3K signaling pathways and activated the PI3K/Akt/NF-κB survival signaling pathway. Our study provided an important insight to reveal the protective molecular mechanism of SAL as a novel drug candidate.

Selective and sensitive fluorescence imaging reveals microenvironment-dependent behavior of NO modulators in the endothelial system / 药物分析学报

Nitric oxide (NO) is a second messenger playing crucial roles in the signaling of a variety of cellular functions. Due to its pathophysiological significance, various NO modulators have been developed to explore NO pathways and some have been used as therapies. These modulators are often used directly to observe pharmacological effects in cell lines, but their actual effect on intracellular NO level is seldom analyzed. Herein, facilitated by a selective and sensitive fluorescence probe, we observed that some NO modulators displayed unexpected behaviors with both NO scavenger carboxy-PTIO and endothelial nitric oxide synthase (eNOS) inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME) failing to decrease intra-cellular free NO level in EA. hy926 cells while NO donor diethylamine-NONOate (DEA?NONOate) and eNOS activator calcimycin (A23187) failing to increase free NO level in human umbilical vein endothelial cell line (HUV-EC-C), although the reagents were confirmed to work normally in the primary human umbilical vein endothelial cells (primary HUVECs) and RAW 264.7 macrophage cells. Further research suggested that these unusual behaviors might be attributed to the cellular microenvironments including both the NO synthase (NOS) level and the endogenous glutathione (GSH) level. Genetically manipulating eNOS level in both cells restores the expected response, while decreasing GSH level restores the ability of DEA?NONOate to increase NO level in HUV-EC-C. These results reveal that the cellular microenvironment has a profound impact on pharmacological effect. Our study suggests GSH as a reservoir for NO in live cells and highlights the value of chemical probes as valuable tools to reveal microenvironment-dependent pharmacological effects.

Detection of Rutin in Sophorae Immaturus Flos with Electrochemical Sensor Assembled with Gold Nanoparticles/Nafion/Multi-wall Carbon Nanotube / 中国实验方剂学杂志

Objective::A new-type electrochemical sensor was developed by electrodepositing gold nanoparticle on the surface of glassy carbon electrode (GCE) modified by Nafion-multi-wall carbon nanotube (MWNTs) by the potentiostatic method, and used for the detection of rutin in practical samples. Method::The electrochemical properties of the sensor in potassium ferricyanide-potassium ferrocyanide system were studied by electrochemical impedance spectroscopy and cyclic voltammetric method, in order to explore the electrochemical performance of the sensor. The electrochemical behavior of rutin on the surface of the sensor was studied by the cyclic voltammetric method, and the factors affecting the performance of the sensor were optimized. The content of rutin was determined by differential pulse voltammetric methods. Result::The optimized conditions were -0.25 V of the deposition potential of gold nanoparticle, 20 s of the deposition time, pH 3.0, and 6 μL of 1 g·L-1 carbon nanotube. Under the conditions, a good linear relationship between reduction peak current and rutin concentration was obtained from 5.0 ×10-9 to 7.0 ×10-7 mol·L-1, and the detection limit was 3.6×10-9 mol·L-1. Conclusion::The developed sensor has a good electrical conductivity and stability, and is highly sensitive and simple for the detection of rutin, with a low detection limit. It has been successfully applied to the detection of rutin in Sophorae Immaturus Flos with the recoveries between 97.6%and 104.4%, which provides a new method for the determination of rutin and a new idea for the quality control of Chinese medicinal materials.

Mechanism of melanoma growth inhibition by exogenous mitochondria / 药学学报

Alterations of mitochondrial structure and function in tumor cells allow cell survival and proliferation under hypoxic and acidic microenvironment. The effect of normal mitochondria on tumor initiation and development remains unknown. In this study, mice were euthanized by rapid cervical dislocation for isolation of hepatic mitochondria, which were injected intravenously to melanoma-bearing mice. This animal experiment had been approved by Southwest University Experiment Animal Ethics Review Committee. The results showed that exogenous mitochondria can significantly inhibit the growth of melanoma. Mitochondria isolated from the liver of young mice had more potent anti-melanoma effect than those isolated from aging mice. The average volume of tumors decreased significantly from 1.35 cm3 to 0.34 cm3, and the average mass of tumors decreased significantly from 0.63 g to 0.22 g. This anti-tumor mechanism might be associated with induction of mitophagy and cell necrosis after the exogenous mitochondria entering the melanoma cells. As mitotherapy can clinically improve somatic cell survival for treatment of pediatric patients with myocardial ischemia, the observed anti-tumor effect of exogenous mitochondria provides a hope for selective tumor treatment.

Enhancement of exposure and reduction of elimination for paeoniflorin or albiflorin via co-administration with total peony glucosides and hypoxic pharmacokinetics comparison / 中国药理学与毒理学杂志

OBJECTIVE Paeoniflorin (PF) and albiflorin (AF) are the major active components of total peony glucosides(TPG)from Paeonia lactiflora Pal,which have many biological activities such as anti-inflammatory, antioxidation and anti-hypertension effects. The drug-drug pharmacokinetic interaction among PF,AF and TPG,the pharmacokinetic comparisons of AF between hypoxia and normoxia,the transport of AF cross the blood-brain barrier cell model and the transport of AF/PF/TPG cross Caco-2 cell model were investigated.METHODS A highly sensitive and rapid UPLC-MS method with multiple-reaction monitoring(MRM)scanning via electrospray ionization(ESI)source operating both in the positive and negative ionization mode was successfully developed and validated for simultaneous quantitation of PF and AF in rat plasma after an oral administration of PF,AF and TPG. RESULTS The validated and developed UPLC-MS/MS method was successfully applied to simultaneously determine the AF and PF concentration in rat plasma and investigate pharmacokinetic interactions after a single intragastrical ad-ministration of PF,AF,co-administration of PF with AF and TPG,respectively.The elimination of both PF co-administered with AF and PF in TPG were slower than those for PF alone and the distribution in the tissues was wider.The combination of PF with AF or TPG could significantly increase the values of the AUC, MRT and t1/2of the drug PF, and reduce the values of CL of PF. From a comparison of the main pharmacokinetic parameters among AF alone, AF combined with PF and AF in TPG, the values of the MRT and t1/2of AF in TPG were greater than that of AF alone,and there were statistically signifi-cant differences in these parameters(P<0.05,P<0.01).It was also noticed that AUC and Cmaxof PF in hypoxia rats were significantly decreased compared with that of normaxia rats, suggesting that there was a decreased exposure of PF in rats under hypoxia. The multiple active components in TPG may lead to DDIs between some P-gp substrates. CONCLUSION The clinical performance of total peony glucosides would be better than that of single constitute. The outcomes of the study are expected to serve as a basis for development of clinical guidelines on total peony glucosides usage.

Retinal blood supply changes after vitrectomy combined with panretinal photocoagulation in PDR patients with anterior segment neovascularization / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate hemodynamic alterations of retrobulbar vessels in proliferative diabetic retinopathy ( PDR) patients with anterior segment neovascularization, before and 3mo after vitrectomy combined with panretinal photocoagulation and to explore the clinical significance. ●METHODS: Color Doppler flow imaging ( CDFl ) was used for measurement of blood flow velocities and resistive indexes ( Rl ) of the ophthalmic artery ( OA ) , short posterior ciliary arteries ( sPCA ) and central retinal artery ( CRA ) in 21 eyes of 21 PDR patients with anterior segment neovascularization. CDFl parameters were obtained before and 3mo after vitrectomy combined with panretinal photocoagulation ( PRP) . ● RESULTS: Peak systolic velocity ( PSV ) and end diastolic velocity ( EVD ) of CRA were significantly increased after surgeries, Rl were decreased significantly (P0. 05). ●CONCLUSION: Vitrectomy combined with panretinal photocoagulation might increase the velocity of CRA, decrease Rl and improve ocular blood supply postoperatively. lt may delay or prevent the process of neovascular glaucoma.

Optimization of reaction conditions and methodological investigation on microtox-based fast testing system for traditional Chinese medicine injection / 中国中药杂志

Vibrio fischeri CS234 was used to establish and optimize microtox assay system, laying a foundation for the application of this method in comprehensive acute toxicity test of traditional Chinese medicine (TCM) injections. Firstly, the Plackett-Burman method was carried out to optimize the factors which would affect Vibrio fischeri CS234 luminescence. Secondly, ZnSO4•7H2O was chosen as reference substance to establish its reaction system with quality control samples. The optimal luminescence conditions were achieved as follows: ①At a temperature of (15±1) ℃, Vibrio fischeri CS234 lyophilized powders were balanced for 15 min, then, 1 mL resuscitation fluid was added and blended for 10 min. 100 μL bacteria suspension was taken to measure the initial luminescence intensity, and then 1 mL resuscitation fluid or test sample was immediately added; after reaction for 10 min, corresponding luminescence intensity was measured again. Resuscitation diluent, osmotic pressure regulator and ZnSO4•7H2O stock solution showed no interference to the determination of Vibrio fischeri CS234 luminescence intensity, so this method was of good specificity. The within-and between-batch precisions of quality controls and the lower limit of quantification (LLOQ) samples were <5% and <10% respectively, while the accuracy ranged between 85.8% and 103.2%. The standard curve equation of ZnSO4•7H2O ranged from 3.86 mg•L⁻¹ to 77.22 mg•L⁻¹ (final concentrations) was y=21.78lnx-15.14, R2=0.998; meanwhile, IC₅₀ of ZnSO4•7H2O to Vibrio fischeri CS234 was 19.90 mg•L⁻¹. ZnSO4•7H2O stock solution and its quality controls were continuously investigated for 120 h and 8 h respectively, and their RSD was lower than 2%, indicating stability at room temperature and 4 ℃ storage conditions. Between pH 4.5-8.0, luminescence intensity of Vibrio fischeri CS234 was controlled within ±10%, and such pH value range could meet the testing needs of the vast majority of traditional Chinese medicine injections. The Vibrio fischeri strain CS234 assay system was specific, stable, sensitive, accurate and adaptable after optimization, so it was suitable for the comprehensive acute toxicity assessment of TCM injections.

Protective effect of α-melanocyte stimulating hormone on glutamate-induced retinal excitotoxicity / 中华实验眼科杂志

Background The excitotoxicity to retinal neurons caused by abnormal elevation of glutamate in retina is a common pathology concomitant with major blind-causing eye diseases.However,an effective approach to protect retinal neurons from glutamate-induced excitotoxicity is still lack.Intraperitoneal administration of α-melanocyte stimulating hormone(α-MSH)has been shown to protect hippocampal neurons from glutamate-induced excitotoxicity.Objective This study was to investigate the protective effect of α-MSH on glutamate-induced excitotoxicity in a chicken embryonic retinal explant culture system.Methods The retinas were isolated from chick embryos at embryonic day 9(E9) and cultured as explants.The explants at 3,5 and 7 days in vitro and the retinas at corresponding embryonic day 12,14 and 16(E12,E14,E16)were collected.The morphology of explant cultures was examined by hematoxylin and eosin staining,and the expression of melanocortin receptors (MCRs)was analyzed by real-time PCR.In the experiment of glutamate-induced excitotoxicity,the retinal explants at 4 days in vitro were treated with glutamate for 48 hours,α-MSH was incubated with the explants 30 minutes before and during the glutamate treatment period.Then the apoptotic cells were detected by TUNEL staining and quantified.The glutamate alone treated-explants and those treated with culture media were included as controls.The expression of glial fibrillary acidic protein(GFAP) at 48 hours after treatment in all retinal explants was analyzed by real-time PCR.Results Hematoxylin and eosin staining showed that the retinal explants exhibited similar morphology to those observed in the retinas from chick embryos at the corresponding developmental stages.The real-time PCR analyses of chick retinas showed that MC1R mRNA level at E9,E12,E14 and E16 was significantly lower than that in post-hatch day 1 (all P=0.000) ;whereas the transcript level of MC5R was significantly increased from E9 to E12 and E14 (both P =0.000),and then gradually decreased from E14 to P1.The expression of these genes showed similar temporal patterns in the retinal explant cultures.TUNEL staining revealed that treatment of the retinal explant cultures with α-MSH substantially and significantly reduced number of apoptotic cells induced by glutamate (P =0.000),which was accompanied by significant suppression of glutamate-induced GFAP up-regulation (P =0.000).Conclusions Application of α-MSH dramatically ameliorated glutamate-induced cell death in retinal explant cultures.This protective effect may be due to α-MSH-mediated suppression of astrogliosis caused by abnormal elevation of glutamate.

Ocular inflammation and pathological characteristics of recurrent experimental autoimmune uveitis in rat / 中华实验眼科杂志

Background Most animal models of experimental autoimmune uveitis (EAU) are single attacked procedure,with a different feature from the natural course of human recurrent autoimmune uveitis.So establishing a recurrent EAU model is necessary for the clinical study on EAU.Objective This study was to establish the recurrent EAU model in rat and investigate the ocular inflammation and pathological manifestation and interleukin-17 (IL-17)expression in the eye.Methods T cells isolated from the spleen and draining lymph nodes of Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 peptide fragments (R16) 10 days earlier were re-stimulated with R16 in vitro and injected into naive syngeneic rats to establish the recurrent EAU models,and the normal Lewis rats were used as controls.The eyes of model rats were then examined daily for clinical signs of uveitis by slit-lamp biomicroscopy and scored Caspi's criteria.The rats were sacrificed 1 month,2,3months after injection respectively,and the retinal sections were prepared for the pathological examination by hemotoxylin & eosin staining.Immunohistochemistry was performed to detect the expression of IL-17 in the retina.Results Adoptive transfer of R16-specific T cells to Lewis rats induced recurrent uveitis.The inflammatory scores on the fourth day,the sixth day,and the inflammatory response disappeared on the tenth day after injection.However,the inflammatory reaction occurred repeatedly 4 or 5 times in the 2-month duration after that,and the right and left eyes of a single recipient showed a different pattern of relapse,and the clinical manifestations of EAU was similar to the natural course to those of human autoimmune uveitis.In the retinal specimens of 1-,2-and 3-month group,the number of inflammatory cells was gradually decreased as the time lapse.Compared with the normal group,the thicknesses of the entire retina,outer nuclear layer and inner nuclear layer decreased with a significant difference among the 4 groups (F=20.46,288.40,4.43,all P=0.00).The number of RGCs in the normal group,1-,2-and 3-month group was 231.27 ± 15.36,225.36 ± 17.79,132.18 ±9.39 and 67.45 ± 11.90,respectively,showing a significant difference among them (F=68.94,P=0.00).Immunohistochemistry showed that the scores of the IL-17 expression in the rat retina were 0.64 ± 0.17,1.92 ± 0.19,1.17 ± 0.23 and 0.83 ± 0.23,showing statistically significant difference (F=64.10,P=0.00).Conclusions The stimulation of R16-specific T cells can induce recurrent EAU in Lewis rat.Th17 is involved in the disease course.

Influence of intravitreal avastin on the expression of cell factors in retinal proliferative membrane in proliferative diabetic retinopathy eye / 中华实验眼科杂志

Background A main cause of visual impairment in proliferative diabetic retinopathy (PDR) is vitreous hemorrhage and retinal detachment due to contraction of fibrovascular membrane.To explore the pathogenic mechanism of fibrovascular membrane is a new target for the prevention and management of PDR.Objective This study was to determine the change in expression of vascular endothelial growth factor (VEGF),connective tissue growth factor(CTGF) and pigment epithelium derived factor(PEDF) in the proliferative membranes of patients with PDR after intravitreal injection of avastin,an anti-VEGF agent.Methods This study was approved by the Medical Ethic Committee of Tianjin Medical College,and written informed consent was obtained from each patient before enrollment.A prospective randomized-controlled study was designed.Twenty-six eyes of 24 patients with PDR scheduled for surgery were enrolled from January to June,2008 in Tianjin Medical College Eye Hospital.The patients were randomized into the simple vitrectomy group and avastin injection combined with vitrectomy group,with matched gender,age and disease duration.1.25 mg (0.05 ml) of avastin was intravitreally injected prior to surgery,and vitrectomy was performed 10 days after injection in the avastin injection combined with vitrectomy group,and only vitrectomy was given in the simple vitrectomy group.Preretinal membrane was collected during the surgery.Expression of VEGF,CTGF and PEDF in the preretinal membranes was assayed by immunochemistry.Results VEGF,CTGF and PEDF were expressed in the cytoplasm.The rate of VEGF expression in the preretinal membranes was 30.77％ in the avastin injection combined with vitrectomy group,showing a significant reduction in comparison with the simple vitrectomy group(100.00％)(U =4.000,P＜0.01).The rate of expression CTGF was remarkable elevated in the avastin injection combined with vitrectomy group compared with the simple vitrectomy group (92.31％ vs.62.54％)(U=7.500,P=0.048).However,no significant difference was found in the expression rate of PEDF between the two groups(100.00％ vs.92.31％) (U =65.500,P =0.299).Conclusions The results suggest that intravitreal injection of anti-VEGF drugs resulted in the decrease of VEGF expression and increased CTGF expression in proliferative membranes from patients with PDR.

Y chromosome microdeletions, chromosome karyotypes and reproductive hormones in patients with azoospermia and severe oligozoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the correlation of azoospermia and severe oligozoospermia with Y chromosome microdeletions, chromosome karyotype and reproductive hormones in male infertility patients.</p><p><b>METHODS</b>We collected semen samples from 63 patients with azoospermia, 49 with severe oligozoospermia and 60 men with normal semen parameters, and determined the incidence of Y chromosome microdeletions, chromosome karyotypes and the levels of reproductive hormones.</p><p><b>RESULTS</b>The incidence rate of Y chromosome microdeletions was 11.11% in the azoospermia and 8.16% in the severe oligozoospermia patients, as compared with 0 in the normal controls (P<0.05). The rate of chromosome abnormalities was 9.52% in the azoospermia group, with statistically significant differences from the severe oligozoospermia and normal control men (both 0) (P<0.05). The levels of FSH and LH were significantly higher in the azoospermia ([20.41 +/- 19.34] IU/L and [11.44 +/- 9.48] IU/L) and the severe oligozoospermia patients ([8.88 +/- 7.04] IU/L and [6.78 +/- 3.85] IU/L) than in the normal males ([3.88 +/- 2.21] IU/L and [4.63 +/- 1.51] IU/L) (P<0.05).</p><p><b>CONCLUSION</b>Examinations of genetics and reproductive hormones are necessary for infertile males with azoospermia and severe oligozoospermia, which may contribute to early diagnosis and treatment.</p>

Circulating microRNAs, novel biomarkers of acute myocardial infarction: a systemic review / 世界急诊医学杂志（英文）

BACKGROUND: MicroRNAs (MiRNA) are a novel class of non-coding RNAs involved in the regulation of gene expression post-transcriptionally by cleavage or translational repression of their specific target miRNAs. Numerous studies have demonstrated that circulating miRNAs are stable and abundant in blood and aberrantly expressed under pathological conditions, including cardiovascular diseases. The implications of circulating miRNAs in acute myocardial infarction have recently been recognized. This review will highlight the potential role of miRNA as a novel class of biomarkers in acute myocardial infarction. METHODS: This systemic review is based on our own work and other related reports. RESULTS: During diseases circulating miRNAs are derived from not only circulating blood cells but also other tissues affected by ongoing diseases. These disease-related miRNAs in the blood can serve as potential biomarkers. CONCLUSION: The circulating miRNAs can be used as novel biomarkers potentially offering more sensitive and specific tests than those currently available for diagnosis of acute myocardial infarction.

Changes of open probability of large conductance Ca(2+)-activated K(+) channels in diabetic coronary smooth muscle cells of rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the changes of open probability (Po) of large conductance Ca(2+)-activated K(+) channel (BK channel) in diabetic coronary smooth muscle cells and elucidate the underlying cellular electrophysiology mechanisms of coronary dysfunction.</p><p><b>METHODS</b>Rat coronary smooth muscle cells were isolated from control group and diabetic group. BK single channel currents were recorded by patch clamp technique in inside-out configuration. Open probabilities were calculated and compared between two groups. After exposure to DHS-1, a specific BK channel activator, Po at 0.2 and 1 µmol/L free Ca(2+) were compared between control and diabetic groups.</p><p><b>RESULTS</b>In the presence of 0.2 µmol/L free Ca(2+), the Po at baseline was significantly lower in diabetic rats than in control rats (0.0032 ± 0.0012 vs. 0.095 ± 0.036, P < 0.05). Cytoplasmic application of DSH-1 significantly increased the Po to 0.335 ± 0.096 (P < 0.05 vs. baseline) in control rats, whereas DSH-1 had no effect in diabetic rats (Po = 0.022 ± 0.018, P > 0.05 vs. baseline). In the presence of 1 µmol/L free Ca(2+), the Po at baseline was also significantly lower in diabetic rats than in control rats (0.210 ± 0.055 vs. 0.458 ± 0.077, P < 0.05). Cytoplasmic application of DHS-1 further robustly enhanced Po to 0.823 ± 0.019 (P < 0.05 vs. baseline) in control rats and to 0.446 ± 0.098 in diabetic rats (P < 0.05 vs. baseline of diabetic rats; P < 0.05 vs. control rats with DHS-1).</p><p><b>CONCLUSION</b>The decrease of Po of BK single channel in coronary smooth muscle cells may be a potential cause for coronary dysfunction in diabetic rats.</p>

Coverage on cataract surgery among adults aged 40 or above in a multiethnic rural district of high altitude area in Gongshan county, Yunnan province, China / 中华流行病学杂志

Objective To estimate the prevalence of cataract and its surgical coverage rate together with the burden related to bilateral cataract-blindness,among adults aged 40 or above in Gongshan county of Yunnan province and to evaluate the current cataract status and the efficacy of local cataract prevention program.Methods Cluster sampling was used.The protocol consisted of personal interview,pilot study,visual acuity checking,measuring the intraocular pressure; slit lamp microscopy and the fundus of the eye examination etc.Cataract was graded clinically using the Lens Opacity Classification System (LOCS) ]Ⅲ.Bilateral cataract-blindness burden,bilateral cataractblindness burden and cataract surgical coverage rate were calculated respectively,using two different criteria.Odds ratios (OR) were compared among different groups regarding age,gender,education,ethnic group and altitude of living area.Results Among the 1236 eligible residents,1116 (90.3％)were enrolled in the present study.The prevalence of cataract was 23.8％ among adults aged 40 or order.When the bilateral best refractive vision ＜3/60 was defined as the blindness criterion,the bilateral cataract-blindness burden showed as 1.3％,and cataract blindness surgical coverage rate was 50.0％.When the bilateral presenting vision ＜ 6/60 was defined as the blindness criterion,the bilateral cataract-blindness burden was 25.0％,and cataract blindness surgical coverage rate was 12.9％.The cataract surgical coverage rates were much lower and the bilateral cataract-blindness burden much higher in women,illiterates,living in high altitude areas and those who were aged 70 or above.Conclusion Cataract blindness was a serious public health problem in aged individuals and illiteracy in the residents of the studied areas.Poor prevention programs on cataract called for urgent action to be taken.

Molecular mechanisms of diabetic coronary dysfunction due to large conductance Ca2⁺-activated K⁺ channel impairment / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Diabetes mellitus is associated with coronary dysfunction, contributing to a 2- to 4-fold increase in the risk of coronary heart diseases. The mechanisms by which diabetes induces vasculopathy involve endothelial-dependent and -independent vascular dysfunction in both type 1 and type 2 diabetes mellitus. The purpose of this study is to determine the role of vascular large conductance Ca(2+)-activated K(+) (BK) channel activities in coronary dysfunction in streptozotocin-induced diabetic rats.</p><p><b>METHODS</b>Using videomicroscopy, immunoblotting, fluorescent assay and patch clamp techniques, we investigated the coronary BK channel activities and BK channel-mediated coronary vasoreactivity in streptozotocin-induced diabetic rats.</p><p><b>RESULTS</b>BK currents (defined as the iberiotoxin-sensitive K(+) component) contribute (65 ± 4)% of the total K(+) currents in freshly isolated coronary smooth muscle cells and > 50% of the contraction of the inner diameter of coronary arteries from normal rats. However, BK current density is remarkably reduced in coronary smooth muscle cells of streptozotocin-induced diabetic rats, leading to an increase in coronary artery tension. BK channel activity in response to free Ca(2+) is impaired in diabetic rats. Moreover, cytoplasmic application of DHS-1 (a specific BK channel b(1) subunit activator) robustly enhanced the open probability of BK channels in coronary smooth muscle cells of normal rats. In diabetic rats, the DHS-1 effect was diminished in the presence of 200 nmol/L Ca(2+) and was significantly attenuated in the presence of high free calcium concentration, i.e., 1 mmol/L Ca(2+). Immunoblotting experiments confirmed that there was a 2-fold decrease in BK-b(1) protein expression in diabetic vessels, without altering the BK channel α-subunit expression. Although the cytosolic Ca(2+) concentration of coronary arterial smooth muscle cells was increased from (103 ± 23) nmol/L (n = 5) of control rats to (193 ± 22) nmol/L (n = 6, P < 0.05) of STZ-induced diabetic rats, reduced BK-b(1) expression made these channels less sensitive to intracellular Ca(2+), which in turn led to enhanced smooth muscle contraction.</p><p><b>CONCLUSIONS</b>Our results indicated that BK channels are the key determinant of coronary arterial tone. Impaired BK channel function in diabetes mellitus is associated with down-regulation of BK-b(1) expression and reduction of the b(1)-mediated BK channel activation in diabetic vessels.</p>

Effect of mesenchymal stem cells on the expression of interleukin-17 in the retina of experimental autoimmune uveoretinitis / 中华实验眼科杂志

BackgroundStudies determined that Th17 cells are important inflammatory cell group in experimental autoimmune uveoretinitis ( EAU ).Interleukin-17 ( IL-17 ),as a marker of Th17,is involved in the occurrence and development of EAU.Mesenchymal stem cells (MSCs) play an immunomodulatory role,mainly by inhibiting the expression of Th17 in a variety of self-autoimmune disease.This is one of the current research focuses.ObjectiveThe present study was to investigate the therapeutic effect of MSCs in EAU and their impact on IL-17 expression in the retina.MethodsMSCs were isolated from the bone marrow of the femurs from 10 SPF Wistar rats and cultured and passaged.The third to fifth generations of cells were used in this experiment.EAU models were induced in 48 6-8 week-old SPF Lewis rats by subcutaneous injection of interphotoreceptor retinoid binding protein (IRBP) R16 peptide emulsified in adjuvant.EAU rats were randomly assigned to the model control group and the MSCs group.MSCs suspension (5×106) of 1 ml was injected via the rat tail vein once a day for 3 consecutive days after immunization,and the same amount of PBS was injected in the model control group in the same manner.Six matched normal Lewis rats were used as the normal control group.The inflammatory response was clinically examined under the slit lamp biomicroscope daily,and the histopathological changes of the retina were examined by hematoxylin and eosin staining on days 9,12,15 and 20.The clinical and histopathological scoring was performed according to the Caspi criteria.Expression of the IL-17 protein in the retina was detected by immunohistochemistry on 9,12,15 and 20days following molding.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by Tianjin Municipality Science and Technology Commission.Results MSCs showed the fusiform in shape and vortex-like growth.Flow cytometry verified that presented the positive expression for CD29 and CD44 but absent expression for CD45 and CD34.The scores of the anterior segment were significantly lower ( U=2.815,P =0.005 ; U =2.768,P =0.006 ; U =2.900,P =0.004 ; U =2.855,P =0.004 ),and the retinal inflammation scores were lower in the MSCs group than the model control group at various time points ( U =2.345,P =0.019 ; U =2.559,P =0.011 ; U =2.166,P =0.030 ; U =2.373,P =0.018 ).Im mnunochemistry showed that the expressions levels of the IL-17 protein (A value) in the rat retina were 26.47±5.68,77.78± 9.65,47.02±6.68 and 26.59±5.94 in the MSCs group on days 9,12,15 and 20,and those in the control group were 45.34±4.63,105.95± 10.74,64.11 ±9.76 and 37.02±6.51,showing a significant reduction ( t =6.305,P =0.000 ; t =4.799,P =0.001 ; t=3.540,P=0.005;t=2.900,P=0.016). ConclusionsMSCs can inhibit the aggravation of EAU and suppress the expression of IL-17 in ocular tissue.

Recognition of anti-VEGF therapy base on the mechanism of VEGF in wet age-related macular degeneration / 中华实验眼科杂志

Age-related macular degeneration (AMD) is the leading cause of visual impairment among older population worldwide,and wet AMD is more threatened to vision because of the choroidal neovascularization.Some physical therapies are thought to destroy the lesions but can not improve the visual acuity.Therefore,anti-VEGF drug therapy is becoming a new approach to the management of wet AMD.Vascular endothelial growth factor(VEGF) is thought to play an important role in the complicated pathogenesis,which can be addressed by disease reduction strategies.Among the anti-VEGF drug therapies,anti-VEGF monoclonal antibodies are proved to maintain and improve visual acuity.Other therapies have been or now being developed for the treatment of neovascular AMD with the goal of inhibiting VEGF.These inhibitors include VEGF receptor decoy aflibercept,small interfering RNA-based therapies (bevasiranib) and tyrosine kinase inhibitors (vatalanib),which could offer the potential for further advances.To completely realize the active mechanism of VEGF in wet AMD is helpful for the rational use of anti-VEGF drugs.