BackgroundSeveral cornea collagen crosslinking methods have been used to treat keratoconus.However,the safety of these methods is dissatisfactory.Glyceraldehyde is a very potent and highly reactive crosslinking agent,with little toxicity,but its effect on corneal biomechanical property is poorly clear.ObjectiveThe aim of this study was to evaluate the biomechanical effects of glyceraldehyde collagen crosslinking on rats cornea.Methods Fifteen clean SD rats were randomly divided into 0.005 mol/L glyceraldhyde group,0.050 mol/L glyceraldhyde group and blank control group.Glyceraldhyde drops was topically administered in the right ryes 2 times per day for consecutive 7 days in the 0.005 mol/L and 0.050 mol/L glyceraldhyde groups,and no any eye drops was used in the blank control group.Seven days later,the rats were sacrificed.Transparency of corneal buttons in these different groups was evaluated.The central corneal strips of 2 mm×6 mm with 2 mm scleral tiasue were obtained for the biomeehanical stress-strain measurement,including ultimate stress ( MPa),ultimate strain (％) and 6％ elastic modulus (MPa).Corneal collagen fibril density was assessed by histological examination under a light microscopy.The use of the animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.ResultsThe words could be clearly displayed transcorneally in all the three groups.When strain was 6％,the stress was (0.463±0.065 ) MPa in 0.005 mol/L glyceraldehyde group,(0.846±0.240) MPa in 0.050 mol/L glyceraldehyde group,both showing a significant increase in comparison with (0.195±0.103 ) MPa of the blank control group (P=0.029,0.000 ).Following the crosslinking treatment,the ultimate stress was significant elevated in 0.050 mol/L glyceraldehydes group compared with the blank control group ( ( 10.759 ± 3.337 ) MPa vs.(5.295± 1.313 ) MPa,P =0.007 ),but no significant change between the 0.005 mol/L glyceraldehydes group and the blank control group ( ( 6.043 ±2.084) M Pa vs.(5.295 ± 1.313 ) MPa,P =0.660 ).Corneal ultimate strain was lower in the 0.005 mol/L glyceraldehyde group and 0.050 mol/L glyceraldehyde group than the blank control group (36.57％ ±3.09％ vs.43.87％ ± 1.89％,P =0.009;28.53％ ±1.89％ vs.43.87％ ± 1.89％,P =0.000).However,significantly increased 6％ elastic modulus were seen in the 0.005 mol/L glyceraldehyde group and 0.050 moL/L glyceraldehyde group compared with the blank control group ( ( 7.718 ± 1.076 ) MPa,( 14.102 ± 4.011 ) MPa vs.( 3.252 ± 1.717 ) M Pa),with statistically significant differences ( P =0.029,0.000).Histological examination showed a increase of collagen fiber density in the 0.050 mol/L glyceraldehyde group.Conclusions Corneal collagen crosslinking induced by glyceraldehyde strengthens biomechanical intensity and increases the density of corneal collagen fiber.But the safety of glyceraldehyde crosslinking for keratoconus needs further study.

Background Arresting the overexpression of vascular endothelial growth factor (VEGF) will be a new approach to the inhibition of neovascularization.RNA interference (RNAi) can inhibit the expression of specific gene,and its application in eye has little interference to other gene expression.Objective This study was to investigate the effect of small interference RNA (siRNA) targeting VEGF on the expression of VEGF and retinal neovascularization in oxygen-induced retinopathy (OIR) model.Methods psi-HITM/enhanced green fluorescent protein (EGFP)/VEGF siRNA was designed and prepared in vitro.Mouse endothelioma (EOMA) were cultured in DMEM without antibiotic and divided into 5 groups.The cells were incubated in DMEM only in the blank control group;while 1 μl of LipofectamineTM 2000 + psi-HITM/EGFP,1 μl LipofectamineTM 2000 + 40,50 or 60 nmol/L of psi-HITM/EGFP/VEGF siRNA was added into DMEM in the negative control group and siRNA groups,respectively.The expression of VEGF mRNA and protein was detected by real time PCR (RT-PCR) and Western blot.The optimal effective concentration of VEGF siRNA was assessed.OIR models were established in 48 7-day-old C57BL/6J mice by raising them at an oxygen concentration of (75±2) ％ for 5 days and then to normal air.The mice were randomized into the model group,null vector group and VEGF siRNA group.1 μl of a mixture of psi-HITM/EGFP or VEGF siRNA (60 nmol/L) and LipofectamineTM 2000 was intravitreally injected,respectively,in the null vector group and VEGF siRNA group.The normal mice were used as the normal control group.Expression of VEGF mRNA and protein in the mouse retinas was detected by RT-PCR and Western blot,respectively,and FITC-dextran stretched retinal preparation was examined to evaluate the neovascularization,and retinal sections were examined to quantify the number of vascular endothelial cell nuclei extending beyond the internal limiting membrane (ILM).Results The in vitro transfection test showed that the expression of VEGF mRNA and protein in the EOMA cells was significantly different among the blank control group,negative control group and 40,50,60 nmol/L VEGF siRNA groups (F =148.890,P ＜ 0.001; F =306.960,P ＜ 0.001),and the expression of VEGF mRNA was lower in different concentrations of VEGF siRNA groups than that in the blank control group (t=73.950,119.890,156.480,all at P＜0.001).Also,the expression of VEGF protein was less in different concentrations of VEGF siRNA groups than that in the blank control group (t =15.452,23.344,42.119,all at P＜0.001).The optimal inhibitory concentration of VEGF siRNA was 60 nmol/L.In vivo study determined that compared to the model group and null vector group,the non-perfusion zones and neovascular net in the retina were decreased,and the number of vascular endothelial cell nuclei extending beyond the ILM was less in the VEGF siRNA group.The relative expression level of VEGF mRNA in the retinas was 1.23±0.18,4.02±0.16,3.98±0.19 and 1.98±0.12 in the normal control group,model group,null vector group and VEGF siRNA group,respectively,with a significant difference among them (F=369.780,P＜0.001),and the relative expression levels of VEGF mRNA in the model group and null vector group were higher than that in the normal control group (t=37.880,37.336,both P＜0.001),and the expression of VEGF mRNA in the VEGF siRNA group declined by 50.8％ (t=10.183,P＜0.001).The difference in the expression levels of VEGF protein also was assayed among the various groups (F=408.980,P＜0.001),and VEGF level in the retina was lowered by 68.0％ in the VEGF siRNA group compared to the model group (t =11.473,P＜0.001).However,VEGF level in the VEGF siRNA group remained at a high level in comparison with the normal control group (t =2.422,P＜0.001).Conclusions Intravitreal injection of VEGF siRNA can attenuate retinal neovascularization by effectively downregulate the expression VEGF mRNA and protein in the retina.

Objective To evaluate the changes of the waveform of the photopic negative response in flash-electroretinogram, visual acuity and central retinal thickness in the treatment of intravitreal injections of bevacizumub. Design Retrospective self-comparative case series. Partidpants 8 subjects (9 eyes) with exudative age-related macular degeneration and 3 subjects (3eyes) with proliferative diabetic retinopathy. Method Evaluation protocol included examinations of the Early Treatment Diabetic Retinopathy study visual acu-ity, visual field, intraocular pressure, fundus fluorescein angiography, optical coherence tomography and flash-electroretinogram. Intravit-real injections of bevacizumab, 1.25 mg (0.05ml), were given under an operating microscope and aseptic conditions. All the subjects were followed-up one month later. Main outcome Measure The amplitudes of PhNR, visual acuity and central retinal thickness. Re-sult At 1 months, the mean amplitudes of PhNR and mean visual acuity in all cases had no obvious change (n=12, P>0.05).The central retinal thickness reduced obviously (n=12, P<0.05), but it was neither significantly correlated with PhNR (r=0.294, P=0.145) nor with visual acuity(r=-0.358, P=0.073). Conclusion The single intravitreal injection of bevacizumab is showed promising in absorption of in-traretinal edema and subretinal fluid in patients with exudative age-related macular degeneration and proliferative diabetic retinopathy, but the changes of visual function (including PhNR) might need further investigation. (Ophthalmol CHN, 2009, 18: 243-246)

OBJECTIVETo investigate the effect of Si' an Capsule combined Enalapril on hypertensive trough/peak ratio (T/P) and circadian rhythm of blood pressure in patients with essential hypertension (EH).

METHODSSixty patients with EH of stage II, III were randomly divided into two groups, the 30 patients in the treated group were treated with Si' an capsule combined Enalapril, and the 30 patients in the control group were treated with Enalapril alone. All patients were given 24-hour ambulatory blood pressure monitoring before and after 4 weeks treatment. T/P of systolic and diastolic blood pressure (SBP & DBP) of each group were calculated. Circadian rhythm of blood pressure was observed at the same time.

RESULTST/P of SBP and DBP in the treated group was 62.4 +/- 7.0% and 66.3 +/- 8.1% respectively, which was obviously higher than that in the control group, 53.3 +/- 6.7% and 60.1 +/- 7.2%, respectively (P < 0.05). The blood pressure circadian rhythm recovering rate in the treated group was 73.3% and in the control group 50%, the difference was insignificant.

CONCLUSIONThe combination therapy of Si' an capsule and Enalapril could lower the blood pressure smoothly and restore the circadian rhythm of blood pressure in EH patients.

Aged ; Antihypertensive Agents ; therapeutic use ; Blood Pressure Monitoring, Ambulatory ; Capsules ; Circadian Rhythm ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Enalapril ; therapeutic use ; Female ; Humans ; Hypertension ; drug therapy ; Male ; Middle Aged ; Phytotherapy

Background Our previous study determined that expressions of matrix metalloproteinases (MMPs) and tissue matrix metalloproteinase inhibitors (TIMPs)change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10％ fetal bovine serum,20％,15％,10％,5％ of drug serum and 8 ml/L epidermal growth factor(EGF) was added into the medium respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 (A value)in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20％,15％,10％,5％ drug serum groups and EGF group(F=466.664,P＜0.05),and that in the 10％ ([9.92±0.14] mg/L) and 20％ ([11.87 ±0.11] mg/L) drug serum groups was significantly lowed in comparison with the control group([16.31±0.10] mg/L)(t=99.974,87.394,P＜0.05).The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different(F=158.168,P＜0.05),with a lower value in the 20％ drug serum group compared with the control group ([3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L) (t =21.991,P ＜ 0.05).Also,the significantly different expressing value of TIMP1 was seen among all the groups (F=183.508,P＜0.05),and expressing value of TIMP1 in the 15％ drug serum group was(1.88±0.06)mg/L,which was lower than(3.20±0.32) mg/L of the control group(t=10.353,P＜0.05).Furthermore,the expressing value of the TIMP3 in the cells was significantly different among the various groups(F=54.503,P＜0.05),and that of the 20％ drug serum group was (1.743±0.065)mg/L and it was significantly higher than (1.54 ± 0.05) mg/L of the control group (t =5.046,P =0.004).However,the expressing value of TIMP3of the 15％,10％ and 5％ drug serum groups was lower than that of the control group,respectively all at(P＜0.05).Conclusions Qijingmingmu soup drug serum at the concentration of 20％ can down-regulate the expressions of MMP1,MMP3,TIMP1 and up-regulate the expression of TIMP3 in human conjunctivochalasis bulbar conjunctival fibroblastsin vitro,which probably plays preventive and therapeutic effects on conjunctivochalasis.

BACKGROUND:Increasing evidence has shown that insulin-like growth factor-1 (IGF-1) plays a regulatory role in bone metabolism and increases bone formation,stimulates osteoblast number and activity,as well as reduces osteoclast differentiation and bone resorption.The mechanism of IGF-1 is an issue of concern.OBJECTIVE:To review the research advance in IGF-1 and the bone metabolism-related indexes in postmenopausal osteoporosis.METHODS:The first author searched the PubMed and CNKI databases between January 2012 and July 2016 using the keywords of "postmenopausal osteoporosis,insulin-like growth factor,bone metabolism,biochemical markers" in English and Chinese,respectively.The repetitive articles were excluded,and 36 eligible articles were enrolled for overview.RESULTS AND CONCLUSION:Bone resorption is increased in postmenopausal woman through the regulation of a variety of cytokines,in which IGF-1 in the blood is combined with IGF binding protein 3,making growth hormone play its biological role.In addition,the growth hormone in the blood directly acts on the adipose tissue,and muscle and bones.Growth hormone exerts a direct effect on bone tissue,promotes osteoblast maturation and differentiation,and enhances the expression of collagen and non-collagen through IGF-1-mediated indirect effect,thus promoting bone formation.The process of bone metabolism is able to reflect the activities of osteoblasts and osteoclasts and the changes of bone matrix and bone mineralization.In vitro experiments show that IGF-1 stimulates the proliferation of osteoblast precursors and differentiate into osteoblasts in a dose-dependent manner,promotes the expression of osteogenic markers such as alkaline phosphatase,type Ⅰ collagen,and osteocalcin,and also stimulates the activity and number of osteoclasts.However,there are few clinical reports and few observation indicators,resulting in a lack of reference range for the detection and treatment of osteoporosis,which needs further exploration.

DNA, Viral ; blood ; isolation & purification ; False Negative Reactions ; Hepatitis B virus ; genetics ; Humans ; Indicators and Reagents ; classification ; Polymerase Chain Reaction ; methods

In order to compare the differences of 35 Menthae Herba samples collected on the market and at producing areas, the contents of six total terpenoids, the essential oil and chromatographic fingerprints were analyzed, which provided evidences for drawing up the commodity specifications and grading criteria of Menthae Herba. GC-MS method was used to analyze the chemical constituents of 35 different samples. The chromatographic fingerprints obtained by using GC were then evaluated by similarity analysis, hierarchical clustering analysis and principal component analysis. The relativity between the content of six terpenoids and the essential oil were studied. In this study, the chemical profiles of 35 samples from different producing areas had significant disparity. All samples collected in the report could be categorized into four chemical types, L-menthol, pulegone, carvone and L-menthone, but the chemical profiles had no relationship with the areas. The chromatographic fingerprints of the samples from different types were dissimilar, while the different producing areas were difficult to be separated. It was indicated that the content of volatile oil was positively correlated with the content of L-menthol and the sum of six total terpenoids. The content of the essential oil, L-menthol and the sum of six total terpenoids of Menthae Herba were considered as one of the commercial specifications and grading criteria. These results in the research could be helpful to draw up the commercial specification and grading criteria of Menthae Herba from a view of chemical information.
Cluster Analysis ; Gas Chromatography-Mass Spectrometry ; Mentha ; chemistry ; Oils, Volatile ; analysis ; Principal Component Analysis ; Terpenes ; analysis

Activin Receptors, Type I ; analysis ; Animals ; Arcuate Nucleus of Hypothalamus ; chemistry ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Immunohistochemistry ; Liver Regeneration ; Protein-Serine-Threonine Kinases ; RNA, Messenger ; analysis ; Rats ; Receptors, Transforming Growth Factor beta ; analysis ; Sodium Glutamate ; Transforming Growth Factor alpha ; analysis ; genetics ; Transforming Growth Factor beta ; analysis ; genetics

AIMTo investigate the role of MAPK in the migration of human coronary artery smooth muscle cells(HCASMC ) into three-dimensional fibrin gels.

METHODSHCASMC were primarily cultured. HCASMC migration was measured with a phase-contrast microscope in the presence or absence of PD98059, SB203580, and SP600125, the inhibitors of ERK, p38, and JNK, respectively. Phosphorylation of ERK, p38 and JNK were analyzed by Western blotting in the presence or absence of PD98059, SB203580 or SP600125.

RESULTSHCASMC that migrated into the three-dimensional fibrin gel exhibited a characteristic elongated spindle-shaped appearance and formed vessel-like structure. The number of migrated HCASMC increased with incubation time and concentration of fibrinogen in the range between 0.8 g/L and 6.4 g/L. Western blot showed that fibrin induced phosphorylation of ERK, p38 and JNK time dependently and PD98059, SB203580 and SP600125 could inhibit their activation, respectively. Migration of HCASMC into the fibrin gels was inhibited by SP600125 20 micromol/L and SB203580 10 micromol/L, respectively. Furthermore, inhibition of SP600125 20 micromol/L had a more profound effect. PD98059 50 ,mol/L, however, failed to influence migration of HCASMC. Hence, migration of HCASMC into the fibrin gels is JNK- and p38-dependent, but not ERK-dependent.

CONCLUSIONFibrin gel induces HCASMC migration into itself by activation of JNK and p38, but not ERK, which may play an important role in pathogenesis of atherothrombosis and restenosis.

Anthracenes ; pharmacology ; Cell Movement ; Cells, Cultured ; Coronary Vessels ; cytology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibrin ; metabolism ; Flavonoids ; pharmacology ; Humans ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; MAP Kinase Signaling System ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Pyridines ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism