Background Wearing of soft contact lens,as one of the important methods to correct ametropia,has been widely applied at present.The clinically deleterious effects of soft contact lens have been reported,but researches about influence of apoptosis of corneal epithelial cells are scant.Objective This study was to detect and compare the ratio of apoptosis of corneal epithelial cells between the patients with contact lens wearing and without contact lens wearing,and explore the influence of soft contact lens wearing for long term on growth of corneal epithelial cells after injury.Methods A retrospective nest case-controlled study was designed.Forty eyes of 20 myopic patients with wearing of soft contact lens for ≥5 years and 40 eyes of 20 myopic patients without wearing of soft contact lens were included in Affiliated Hospital of Guiyang Medical College.All the patients received the off-flap epipolis laser in situ keratomileusis (Epi-LASIK) from 2010 October to 2011 June.The corneal flaps were collected during the surgery.The apoptosis of corneal epithelial cells was detected by flow cytometry(FCM),and the difference of apoptotic proportion between the two groups was compared using independent sample t test.Results The FCM scatter plot showed that compared to the without contact lens wearing group,the early stage of apoptotic cells with high staining annexin V and low staining proaium iodide (PI) were much more,and survival cells with low staining annexin V and PI were less,and necrotic and late stage of apoptotic cells with high staining of annexin V/PI were more in the with contact lens wearing group.The early stage of apoptosis proportion of corneal epithelial cells was (11.23 ± 5.31)％ in the with contact lens wearing group,and that in the without contact lens wearing group was (7.31± 5.43)％,showing a significant difference between them (t=2.754,P=0.008).Conclusions A long term wearing of contact lens induce more apoptosis of corneal epithelial cells,which can result in a disorder of structure and function of ocular surface during the wound healing stage.

Objective To explore the measures to prevent recurrent laryngeal nerve (RLN)injury during thyroid surgery.Methods The clinical data of 223 patients undergone thyroid surgery were retrospectively analyzed.Among the 223 surgeries,69 sides were undergone regional protection act of RLN and 191 sides were performed RLN exposure.Results There were 2 cases of RLN injury from the regional protection operation of RLN,including 1 case of temporary nerve injury which could be resulted from surgery clamp and 1 case of permanent nerve injury which might be caused by mistaking ligation during surgery.There was only 1 case of temporary nerve injury in RLN exposure procedure which was probably caused by the postoperative nerve edema and was recovered 2 months after the operation.The total RLN injury rate was 1.35％.Conclusion For benign thyroid lesions and non-dorsal lesions or during partial excision of the gland,the regional protection of RLN is helpful to prevent RLN injury.In cases with dorsal lesions of thyroid or contralateral RLN injury,or during lobe subtotal resection,lobe resection and reoperation,exposing RLN to prevent injury is necessary.Taking different approaches based on the profiles of lesions and surgical procedures to prevent RLN injury can significantly reduce the risk of RLN injury.

AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.

Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1％ R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P＜ 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P＜0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P＜ 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.

Health Personnel ; psychology ; Humans ; Logistic Models ; Multivariate Analysis ; Professional Competence ; statistics & numerical data ; Stress, Psychological ; epidemiology

To investigate the protective effect of preconditioning with hyperoside ( Hyp) against myocardial ischemia-reperfusion injury (MIRI) in rats and the role of PI3K/Akt signaling pathway. MIRI was established by ligation of left anterior descending coronary artery for 30 min followed by reperfusion for 120 min in rats. Male SD rats were randomly divided into five groups: sham group,model group (MIRI),Hyp preconditioning group(Hyp), Hyp preconditioning + LY294002 (a PI3K/Akt signaling pathway inhibitor) group (Hyp + LY), and LY294002 group (LY). At the end of reperfusion, hemodynamic parameters were recorded as left ventricular systolic pressure (LVSP) , left ventricular end-diastolic pressure ( LVEDP) and maximal rate of increase and decrease of left ventricular pressure (± dP/dt(max)). Myocardial infaret size, the oxidative stress markers, myocardial enzymes indicators and inflammatory factors were also analyzed. The expressions of Akt, p-Akt, Bax and Bcl-2 proteins was detected by using Western blot method. The results showed that Hyp preconditioning remarkably improved cardiac constriction and relaxation function, reduced myocardial infarct size and enhanced the activities of oxidative stress markers about correlated to MIRI, such as superoxide dismutase (SOD), catalase (CAT) and glutathione-peroxidase (GSH-Px) and decreased the contents of malondialdehyde (MDA) as compared with MIRI group. Simultaneouly, the levels of myocardial enzymes, i. e. creatine kinase ( CK) and creatine kinase MB isoenzyme (CK-MB), and inflammatory factors, for instance tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were decreased. Hyp pretreatment apparently restrained myocardial apoptosis as evidenced by decreasing the level of Bax expression, increasing the levels of phosphorylation of Akt and Bcl-2 expression. These effects were inhibited by LY294002, a blocker of PI3K/Akt signaling pathway. These findings indicated that the cardioprotection of Hyp preconditioning against MIRI may be related to activating PI3K/Akt signaling pathway, upregulating the expression of BCL-2 protein and down-regulating the expression of Bax protein.
Animals ; Creatine Kinase ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Heart ; drug effects ; Humans ; Interleukin-6 ; genetics ; metabolism ; Ischemic Preconditioning, Myocardial ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; drug therapy ; enzymology ; genetics ; prevention & control ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Quercetin ; administration & dosage ; analogs & derivatives ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; genetics ; metabolism

To investigate the effects and potential mechanisms of hyperoside (Hyp) on the vascular endothelium function in middle cerebral artery (MCA) ex vivo in rats. Isolated arterial segments from MCAs of rats were used for surveying vasomotoricity in a pressurized chamber. Transmembrane potential was recorded by using glass microelectrodes to evaluate hyperpolarization. Hyp (1 x 10(-6)-1 x 10(-4) mol . L-1) was utilized to observe the effect on 1 x 10(-7) mol . L-1 U46619-preconstricted MCA in rats. The results showed that 1 x 10(-6)-1 x 10(-4) mol . L-1 Hyp significantly induced concentration-dependent vasodilatation and hyperpolarization, leading to the maximal diastolic ratio of (73. 2 ± 6. 1)% and maximal changes in membrane potentials of (-13. 2 ± 2. 2) mV. Hyp still elicited vasorelaxation and hyperpolarization by removal of endothelium in MCA of rat, which was notably attenuated as compared with vascular endothelium-intact group (P <0. 01). In the MCAs preconstricted by U46619 (1 x 10(-7) mol . L-1), Hyp (1 x 10(-6)-1 x 10(-4) mol . L-1) produced concentration-dependent vasorelaxation and hyperpolarizition that were partially attenuated by 3 x 10(-5) mol . L-1 L-NAME(a NOS inhibitor) plus 1 x 10(-5) mol . L-1 PGI2 ,(a synthetase inhibitor). The residual effects were further decreased by 1 x 10(-3) mol . L-1 TEA (an inhibitor of Ca2+-activated potassium channel) or 1 x 10(-5) mol . L-1 PPG (a blocker of endogenous H2S synthese-CSE). Similarly, 1 x 10(-5)-1 x 10(-3) mol . L-1 NaHS (a donor of exogenous H2S) or 1 x 10(-5)-1 x 10(-3) mol . L-1 L-Cys (the substrate of endogenous H2S synthesis) obviously evoked dose-dependent vasodilatation and hyperpolarization of MCA in rats. These findings indicated that Hyp may induce endothelium-dependent and endothelium-independent responses. And the endothelium-dependent vasodilatation may be related to the increases of endogenous H2S that has been promoted Hyp in the endotheliocyte of MCAs, and activated Kca and opening of Kca channels, resulting in the hyperpolarization of vascular smooth muscle cell membrane and subsequent reduction of Ca2+ influx and vasodilation.
Animals ; Endothelium, Vascular ; drug effects ; Enzyme Inhibitors ; pharmacology ; Female ; Male ; Middle Cerebral Artery ; Muscle, Smooth, Vascular ; drug effects ; Nitric Oxide ; metabolism ; Potassium Channels ; metabolism ; Quercetin ; analogs & derivatives ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfides ; metabolism ; Vasodilation ; drug effects ; Vasodilator Agents ; pharmacology

Uridine diphosphate (UDP)-GalNAc : polypeptide N-acetylgalactosaminyltransfemse (ppGalNAcT) catalyzes the initial step in mucin type O-glycosylation in the Golgi apparatus. Here generation and characterization of a polyclonal antibody to human ppGalNAcT2 were described. The subcellular location of ppGalNAeT2 in SGC7901 cell line was investigated using Western blot analysis of fractionated cell extracts and confocal microscopy with this antibody and two Golgi markers: Golgi SNARE (soluble N-ethylmalemide-sensifive factor attachment protein receptor) of 28 ku (GS28) and trans-Golgi network (TGN) 38, markers for the c/s- and trans-Golgi apparatus, respectively. Morphometric analyses indicated that ～60% of the ppGalNAcT2 signal colocalized with the GS28, while～36% of the c/s-Golgi marker colocalized with the ppGalNAeT2. Approximately 34% of the ppGalNAcT2 signal colocalized with the TGN38, whereas 38% of the trans-Golgi marker colocalized with the ppGalNAcT2. The results provide unequivocal evidence for the location ofppGalNAcT2 within the Golgi apparatus, and further highlight the importance of this organelle in the initiation of O-linked glycosylation.

OBJECTIVETo investigate the triterpenoids from root of Achyranthes bidentata in Henan.

METHODSephadex, normal-and reversed-phase column chromatographies were applied for the isolation and purification. The structure determinations were performed by means of physiochemical properties, MS and NMR data analyses.

RESULTSeven compounds were isolated from the water soluble fraction in root of A. bidentata, and determined as achyranthoside A (1), achyranthoside E (2), momordin Ib (3), chikusetsusaponin IVa (4), chikusetsusaponin IVa methyl ester (5), chikusetsusaponin V (6), chikusetsusaponin V methyl ester (7).

CONCLUSIONCompounds 1 and 2 were isolated from the natural resources for the first time.

Achyranthes ; chemistry ; Molecular Conformation ; Molecular Structure ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective To examine the association between internet addiction and suicidal behaviors in adolescences and to explore whether the association could be partly or fully accounted by depression or/and behaviors related to the impulsity. Methods A total of 3507 urban adolescent students in Hefei were administered to complete the questionnaire concerning interact addiction disorder(IAD), suicidal behaviors during the 12 months preceding the survey, behaviors related to the impulsity, Center for Epidemiologic Studies Depression Scale and early childhood adversities,respectively. Hierarchical logistic regressions were used to evaluate the associations and possible roles of depression and behaviors related to the impulsity between internet addition and suicidal behaviors after adjustment for confounding factors. Results Of the 3507 participants, 5.2% were diagnosed as IAD, 27.4% reported suicidal ideation during the 12 months preceding the survey, with another 9.5% had a plan and 2.6% had an attempt. Internet addition was associated with suicidal ideation(OR=1.79, 95%CI: 1.32-2.44), plan(OR=2.32, 95%CI: 1.57-3.42), and attempt(OR=2.08, 95%CI:1.03-4.22). After adjustment for potential confounding factors, the association between internet addition and attempt was insignificant. When depression and behaviors related to the impulsity were entered into Hierarchical regression respectively, the associations between internet addition, suicidal ideation and plan were substantially reduced. While internet addition was enter into Hierarchical regression, the associations between depression, behaviors related to the impulsity, suicidal ideation and plan were not reduced. Conclusion Adolescent intemet addition seems to be associated with suicidal ideation and plan, and the association is possibly mediated by depression and behaviors related to the impulsity.