BACKGROUNDCataracts is considered be formed because of an abnormal glucose metabolic pathway or oxidative stress. We explored the damaging role of ONOO- and antagonism of cholecystokinin octapeptide-8 (CCK-8) in diabetic cataractal rat lenses.

METHODSA diabetic cataractal animal model was established by peritoneal injection of streptozotocine (STZ). Thirty-six normal SD rats were taken as control group; seventy-two were given STZ (45 mg/kg) and then divided into STZ group and CCK-8 group (peritoneal injection CCK-8). STZ induced diabetic rats were treated with CCK-8 for 60 days. Lenses were examined with slit lamp at 20, 40 and 60 days. Immunofluorescent staining and Western blot analysis were used for determining nitrotyrosine (NT, a marker for ONOO-). PT-PCR and gene array analysis were used for determining the expression of inducible nitric oxide synthetase mRNA (iNOS mRNA) in lens epithelium (LEC).

RESULTSSTZ group rats developed lens opacity by 20 days that reached a high level by 60 days after STZ injection. CCK-8 group rats delayed the cataract formation. CCK-8 group rats delayed the cataract formation. There was no distinct expression of NT and iNOS mRNA in control group. In STZ group, there were distinct expression of NT and upregulation of iNOS mRNA; however, CCK-8 group showed weak expression of NT and downregulation of iNOS mRNA.

CONCLUSIONSNT, which may be a new form of oxidative stress, was expressed in diabetic rat LEC although CCK-8 could reverse NT damage in LEC. The results suggested that CCK-8 might be a useful therapeutic agent against diabetic cataract. The antagonizing mechanism of CCK-8 may be related to direct antagonism of ONOO- as well as its inhibition of the expression of iNOS mRNA for production of NO and therefore decrease in the formation of ONOO-.

Animals ; Blotting, Western ; Cataract ; etiology ; prevention & control ; Diabetes Mellitus, Experimental ; complications ; Fluorescent Antibody Technique ; Male ; Nitric Oxide Synthase Type II ; genetics ; Oxidation-Reduction ; Peroxynitrous Acid ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sincalide ; pharmacology ; Streptozocin ; Tyrosine ; analogs & derivatives ; genetics

The present study was designed to observe if puerarin decreases lens epithelium cell (LEC) apoptosis induced partly by peroxynitrite (ONOO(-)). One hundred and eight rats were randomly divided into control group (n=36), streptozotocin (STZ) group (n=36) and STZ + puerarin group (n=36). The rats in the control group intraperitoneally (i.p.) received 0.5 ml of saline. The rats in STZ group and STZ + puerarin group received intraperitoneal injection of STZ (45 mg/kg). Three days later, the rats in STZ + puerarin group were given puerarin (140 mg/kg per day, i.p.). On days 20, 40 and 60 of the experiment, morphologic changes of lenses were observed with slit lamp. Then the animals were sacrificed for further analysis. The amount and percentage of apoptotic LECs were determined by flow cytometry. Nitrotyrosine (NT, the foot print of ONOO(-)) was examined by immunohistochemistry. Apoptosis-related genes (iNOS, etc.) were analyzed by gene array. The results showed that in the control group, all the lenses were clear. In STZ group, gradually severe opacity of the lens was observed on days 20, 40 and 60. But in STZ + puerarin group, mild opacity of the lens was observed on day 20 and more severe on day 40, but markedly decreased on day 60. In the control group, mild apoptosis of LECs was observed. In STZ group, time-dependent increase in apoptosis of LECs was observed. In STZ + puerarin group, mild apoptosis of LECs was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. There was no expression of NT in the lens in the control group, but an increased expression of NT in STZ group. In STZ + puerarin group, mild expression of NT was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. There was no expression of iNOS in the lens in the control group, but continuous up-regulation of iNOS expression in STZ group. In STZ + puerarin group, mild expression of iNOS was observed on day 20, significantly increased on day 40, but markedly decreased on day 60. Except the changes of iNOS related to NO production, the other apoptosis-related genes, including BCL-2 and SOD were down-regulated, while NF-kappaB and TNFR1-FADD-caspase signal transduction way were up-regulated in STZ group. The results were opposite in STZ + puerarin group and the control group. These findings show that NT is expressed in diabetic rat lens, which proves that LEC apoptosis in diabetic lens is partly induced by ONOO(-) which may be a new oxidative damage way to form cataract. Puerarin partly decreases LEC apoptosis induced by ONOO(-) and is a potential medicine for therapy of diabetic cataract. The mechanism of puerarin dealing with diabetic cataract may be related to its direct inhibition of LEC apoptosis and antagonism of ONOO(-) in diabetic rats.
Animals ; Apoptosis ; Cataract ; chemically induced ; Diabetes Mellitus, Experimental ; Epithelial Cells ; drug effects ; Isoflavones ; pharmacology ; Lens, Crystalline ; cytology ; Nitric Oxide Synthase Type II ; metabolism ; Peroxynitrous Acid ; Rats ; Tyrosine ; analogs & derivatives ; metabolism

OBJECTIVETo evaluate the clinical outcome among three different laser in situ keratomileusis (LASIK) ablations: Q-factor customized ablation (aberration smart ablation, ASA), wave-front guided ablation (WASCA) and ablation under wave-front guiding plus iris recognition system (IR+WASCA).

METHODSThis prospective study comprised 96 eyes of 96 patients, and they were randomly divided into three groups: 30 patients in ASA group, 32 in WASCA group, and 34 in IR+WASCA group. There were no any statistical differences in spherical equivalent (SE), age, sex, pupil diameter, higher-order aberrations (HOA) among three groups preoperatively. Wave-front analysis was performed before and 1, 3 months after operation.

RESULTAll patients got an uncorrected visual acuity (UCVA) ≥ 0.8 1 and 3 months after operation. The residual SE was in ± 0.50D both at 1 and 3 months after surgery. There was no statistical difference in SE value, HOA, change of HOA, and coma aberration postoperatively among three groups. Horizontal coma (Z3¹) aberration took the majority of coma. HOA, total coma aberrations and spherical aberration increased postoperatively (P = 0.000), but without significant difference between 1 month and 3 months after surgery.

CONCLUSIONThat three customized LASIK are all effective, safe, accurate and stable; meanwhile WASCA may have better UCVA than the other two groups postoperatively.

Follow-Up Studies ; Humans ; Keratomileusis, Laser In Situ ; methods ; Myopia ; surgery ; Prospective Studies ; Treatment Outcome

OBJECTIVETo observe the effects of forearms devascularization on the volt-ampere (V-A) characteristics of Neiguan (PC 6) and Taiyuan (LU 9) so as to investigate the relationship among functions of acupoints, qi and blood in human body and V-A characteristics of acupoints.

METHODSEither forearms of 23 healthy volunteers were devascularized and the changes of V-A characteristics in ipsilateral Neiguan (PC 6) and Taiyuan (LU 9) in normal and devascularization statuses were detected by home-made intelligent V-A detecting system.

RESULTSCompared with the normal status, both range-increasing and range-decreasing V-A areas of Neiguan (PC 6) and Taiyuan (LU 9) were significantly increased in devascularization status (P < 0.05, P < 0.01), and the change rates of these two V-A areas of Neiguan (PC 6) were larger than those of Taiyuan (LU 9) (both P < 0.05). The inertia areas of Neiguan (PC 6) but not Taiyuan (LU 9) became significantly larger after devascularization (P < 0.01).

CONCLUSIONThe changes in V-A characteristics of Neiguan (PC 6) and Taiyuan (LU 9) can reflect the alteration of local blood supply, and Neiguan (PC 6) is more sensitive to changes of blood flow than Taiyuan (LU 9).

Acupuncture Points ; Adult ; Blood Circulation ; Electrophysiology ; Female ; Forearm ; blood supply ; physiology ; Humans ; Male ; Young Adult

Using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, macroporous adsorbent resin, and reversed-phase HPLC, 115 compounds including diterpenes, sesquiterpenes, treterpenes, coumarins, lignans, fatty acid derivatives, and simple aromatic derivatives were isolated from an ethanol extract of branch of Fraxinus sieboldiana (Oleaceaue), and their structures of the compounds were elucidated by spectroscopic methods including 1 D, 2D NMR and MS techniques. Among them, 41 compounds were new. In previous reports, we have been described the isolation, structure elucidation, and bioactivities of the 41 new compounds and 22 known orii including 8 coumarins, 4 phenolic and 12 phenylethanoidal glycosides. As a consequence, we herein reported the isolation and structure elucidation of the remaining 50 known compounds including 8- hydroxy-12-oxoabieta-9(11),13-dien-20-oic 8, 20-lactone(1), 6beta-hydroxyfcrruginol(2),(+)-pisiferic acid(3), (+)-pisiferal(4),(+)-7-dehydroabiet6none(5), 1-oxomiltirone(6), subdigitatone(7), linarionoside B(8), (9S)-linarionoside B(9), (3R,9R)-3-hydroxy-7,8-dihydro-beta-ionol 9-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside(10), ursolic acid(11), betulinic acid(12), euscaphic acid(13), (+)-syringaresinol(14), (+)-fraxiresinol(15), (+)-1-hydroxysyringaresinol(16), pinoresinol(17), medioresinol(18), 8-acetoxypinoresinol(19), epipinoresinol(20), (-)-olivil(21), (+)-cyclo-olivil(22), 3,3'-dimethoxy-4,4',9-trihydroxy-7,9'-epoxylignan-7'-one(23),(+)-1-hydroxypinoresinol 4'-O-beta-D-glucopyranoside (24), (+)-1-hydroxypinoresinol 4"-O-beta-D-glucopyranoside(25),(+)-syringaresinol O-beta-D-glucopyranoside (26), liriodendrin (27), ehletianol D(28), icariside E5(29) (-)-(7R, 8R)-threo-1-C-syringylglycerol(30),(-)-(7R, 8S)-erythro-guaiacylglycerol (31),(-)-(7R, 8R)-threo-guaiacylglycerol(32), 3-(4-beta-D-glucopyranosyloxy-3-methoxy)-phenyl-2E-propenol(33),2,3-dihydroxy-l-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(34), 2,3-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-1-propanone (35), 3-hydroxy-l-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(36), omega-hydroxypropioguaiacone(37), sinapyladehyde(38), trans-p-hydroxycinnamaldehyde(39), syringic acid(40), vanilic acid(41), vanillin(42), 4-hydroxy-benzaldehyde (43), (24R)-24-ethyl-5alpha-cholestane-3beta,5,6beta-triol(44), beta-sitosterol(45), daucosterol(46), 2,6-dimethoxy-I,4-benzoquinone(47), 2,6-dimethoxy-pyran-4-one(48), 1-(beta-D-ribofuranosyl)uracil(49), and mannitol(50). Compouds 1-7,12,18,28-37,44 and 48 were obtained from the genus Fraxinus for the first time.
Fraxinus ; chemistry ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Plant Extracts ; analysis

Acupuncture ; standards ; Acupuncture Therapy ; standards ; China ; Complementary Therapies ; standards ; Guideline Adherence ; Herpes Zoster ; diagnosis ; therapy ; Humans ; Male ; Practice Guidelines as Topic ; standards ; Risk Assessment ; Severity of Illness Index

OBJECTIVETo study the molecular genetic mechanism and genetic diagnosis of pyruvate dehydrogenase complex deficiency (PHD), and to provide a basis for genetic counseling and prenatal genetic diagnosis of PHD.

METHODSPolymerase chain reaction (PCR) was performed to amplify the 11 exons and exon junction of the PDHA1 gene from a child who was diagnosed with PHD based on clinical characteristics and laboratory examination results. The PCR products were sequenced to determine the mutation. An analysis of amino acid conservation and prediction of protein secondary and tertiary structure were performed using bioinformatic approaches to identify the pathogenicity of the novel mutation.

RESULTSOne novel duplication mutation, c.1111_1158dup48bp, was found in the exon 11 of the PDHA1 gene of the patient. No c.1111_1158dup48bp mutation was detected in the sequencing results from 50 normal controls. The results of protein secondary and tertiary structure prediction showed that the novel mutation c.1111 _1158dup48bp led to the duplication of 16 amino acids residues, serine371 to phenylalanine386, which induced a substantial change in protein secondary and tertiary structure. The conformational change was not detected in the normal controls.

CONCLUSIONSThe novel duplication mutation c.1111_1158dup48bp in the PDHA1 gene is not due to gene polymorphisms but a possible novel pathogenic mutation for PHD.

Amino Acid Sequence ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Protein Conformation ; Pyruvate Dehydrogenase (Lipoamide) ; chemistry ; genetics ; Pyruvate Dehydrogenase Complex Deficiency Disease ; genetics

BACKGROUNDIn vivo quantification of choroidal neovascularization (CNV) based on noninvasive optical coherence tomography (OCT) examination and in vitro choroidal flatmount immunohistochemistry stained of CNV currently were used to evaluate the process and severity of age-related macular degeneration (AMD) both in human and animal studies. This study aimed to investigate the correlation between these two methods in murine CNV models induced by subretinal injection.

METHODSCNV was developed in 20 C57BL6/j mice by subretinal injection of adeno-associated viral delivery of a short hairpin RNA targeting sFLT-1 (AAV.shRNA.sFLT-1), as reported previously. After 4 weeks, CNV was imaged by OCT and fluorescence angiography. The scaling factors for each dimension, x, y, and z (μm/pixel) were recorded, and the corneal curvature standard was adjusted from human (7.7) to mice (1.4). The volume of each OCT image stack was calculated and then normalized by multiplying the number of voxels by the scaling factors for each dimension in Seg3D software (University of Utah Scientific Computing and Imaging Institute, available at http://www.sci.utah.edu/cibc-software/seg3d.html). Eighteen mice were prepared for choroidal flatmounts and stained by CD31. The CNV volumes were calculated using scanning laser confocal microscopy after immunohistochemistry staining. Two mice were stained by Hematoxylin and Eosin for observing the CNV morphology.

RESULTSThe CNV volume calculated using OCT was, on average, 2.6 times larger than the volume calculated using the laser confocal microscopy. The correlation statistical analysis showed OCT measuring of CNV correlated significantly with the in vitro method (R 2 =0.448, P = 0.001, n = 18). The correlation coefficient for CNV quantification using OCT and confocal microscopy was 0.693 (n = 18, P = 0.001).

CONCLUSIONSThere is a fair linear correlation on CNV volumes between in vivo and in vitro methods in CNV models induced by subretinal injection. The result might provide a useful evaluation of CNV both for the studies using CNV models induced by subretinal injection and human AMD studies.

Animals ; Choroidal Neovascularization ; pathology ; physiopathology ; Disease Models, Animal ; Fluorescein Angiography ; Humans ; Mice ; Mice, Inbred C57BL ; Tomography, Optical Coherence

Adult ; Colitis, Ulcerative ; Colon ; Humans

Objective:To explore the anti-liver cancer potential of Fufang Biejia Ruangan Pian （FBRP） and its compatibility characteristics from a network perspective， so as to provide a theoretical basis for the clinical repositioning of FBRP. Method:Three self-pairs of cancer and para-cancerous tissue samples were collected from three patients with primary liver cancer， and the whole genome expression profiling chip was used to detect the differential genes related to the development and progression of liver cancer. After collecting the phenotype-related genes and the candidate targets of the corresponding prescriptions of FBRP from The Encyclopedia of Traditional Chinese Medicine （ETCM） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） V2.0， the "differentially expressed genes related to liver cancer development-candidate targets of FBRP efficacy substance group" interaction network was constructed according to the interaction information between the above-mentioned differentially expressed genes related to liver cancer and the candidate targets of the FBRP efficacy group， and then the major network nodes were screened. After that， the enrichment analysis of the pathway was performed in order to explore the biological basis of various pharmacological efficacy groups of FBRP， including Xiaozheng Sanjie group （Trionycis Carapax and Curcumae Rhizoma）， Buxue Huoxue group （Paeoniae Radix Rubra， Angelicae Sinensis Radix and Notoginseng Radix et Rhizoma）， Yiqi Jianpi group （Codonopsis Radix and Astragali Radix）， Yuyin Yanggan group （Placenta Hominis and Cordyceps） and Qingre Jiedu group （Isatidis Radix and Forsythiae Fructus）. Result:The major network targets of the five efficacy groups may be involved into several common pathways but also associated with some special pathological processes. Those common pathways mainly contained the regulation of nervous system， the balance of immune-inflammatory system， the regulation of energy metabolism of various substances and cancer-related pathways， while the point was also reflected by the follows：①The regulating effects of Xiaozheng Sanjie group and Yiqi Jianpi group were summarized as promoting Qi circulation and relieving depression and replenishing Qi-blood， benefiting spirit. Buxue Huoxue group may also participate in the regulation of promoting Qi circulation and relieving depression and Yuyin Yanggan group may participate in the regulation of replenishing Qi-blood and benefiting spirit. ②The regulatory effects of the Xiaozheng Sanjie group and the Yuyin Yanggan group were summarized as essence， Qi and blood supplement. Buxue Huoxue group focused on the improvement of the immune-circulatory system. Qingre Jiedu group mainly regulated the balance of immune-inflammatory system by acting on T cell receptor signaling pathway. ③Yiqi Jianpi group was demonstrated to show the effects on various material and energy metabolisms. Yuyin Yanggan group exerted effects on lipid metabolism， carbohydrate metabolism， protein metabolism and hormone metabolism. Qingre Jiedu group was also involved into metabolism of nucleotide and hormone. ④In the aspect of alleviating the pathological changes of cancer， the regulatory effects of the five efficacy groups on cell cycle and other functions could be summarized as dispelling pathogenic factors. ⑤The whole prescription focused on the anti-liver cancer potential of FBRP as a whole， while each efficacy group emphasized that each efficacy group had its own functional characteristics. The two network analysis models complemented and verified each other. Conclusion:FBRP has the anti-hepatoma potential. By revealing the biological connotation of its efficacy and the rationality of the compatibility， the regulation mechanism of FBRP to correct the imbalance network of inflammation and cancer in liver is clarified， which can provide the possibility and biological basis for FBRP to increase the clinical indications for the prevention and treatment of liver cancer.