AIM: Using optical coherence tomography angiography (OCTA) to observe the changes and clinical significance of macular blood flow density in patients with diabetic retinopathy (DR).METHODS: Totally 47 eyes (28 patients) with diabetic retinopathy (DR) were enrolled in the DR group.According to the international clinical grading criteria of diabetic retinopathy, 30 eyes (19 patients) with non-proliferative diabetic retinopathy were classified as the NPDR group, and 17 eyes (11 patients) with proliferative diabetic retinopathy were classified as PDR group.A total of 46 (27 subjects) healthy eyes with matched age were enrolled in the control group.All the subjects underwent the 3mm×3mm scanning of macular retina by optical coherence tomography angiography (OCTA), obtaining 4 levels of macular blood flow density map.The macular blood flow density at 3 levels, including superficial retinal layer, deep retinal layer and choroidal capillaries layer, were measured.RESULTS: The macular blood flow density of superfical retinal layer, deep retinal layer and choroidal capillaries layer in DR group were 0.4963±0.0840, 0.4798±0.0801 and 0.5290±0.0528, respectively.Among them, the blood flow density of each layer were 0.5064±0.0843,0.4983±0.0766,0.5345±0.0529, respectively, for the NPDR group, and were 0.4786±0.0830, 0.4473±0.0778,0.5192±0.0526, respectively, for the PDR group.For the control group, the density of each layers were 0.5919±0.0704, 0.6301±0.0527, 0.5691±0.0169, respectively.The macular blood flow density was significantly different in the superficial retinal layer, deep retinal layer and choroidal capillary layer between the control group and the NPDR group, as well as the PDR group and the DR group (total P<0.001).Statistically significant difference was found between the NPDR group and the PDR group in the deep retina layer (P=0.029), but not in the superficial retina layer and choroid capillary layer (P=0.236, 0.268).CONCLUSION: Compared with the control group, the macular blood flow density of superficial retinal layer, deep retinal layer and choroidal capillary layer in the patients with diabetic retinopathy decreased significantly.It indicated that the macular ischemia existed in both retina and choroid.By quantitatively measurement of the macular blood flow, OCTA may be used for monitoring the progression of diabetes, and early detection of diabetic retinopathy.

Background Ginkgolide B (GB) has been proved to have neuroprotective and anti-apoptotic effects and can effectively inhibit apoptosis of retinal photoreceptor cells.But the high hydrophobic feature and low bioavailability of GB limit its clinical application.Self microemulsifying drug delivery system (SMEDDS) can effectively improve the infusibility drug dissolution and bioavailability in the retina.Objective This study was to investigate the pharmacokinetics and drug-time change of GB-loaded SMEDDS in retina.Methods Eighty SD rats were randomized into 2 groups,2.5％ GB(40 mg/kg) of SMEDDS or GB suspension(0.1％ DMSO dissolve) were gastrically given respectively in two groups.The rats were sacrificed and retinas were isolated 15,30,45 minutes and 1 hour,2,4,8,12 hours to prepare the retinal suspension.The content of GB in retina was assayed with high performance liquid chromatography-electrospray ionization-(1) (1)ss spectrum (HPLC-ESI-MS) and contrasted with standard curve.Practical drug dynamics program 3p87 was used to detect the pharmacokinetics parameters.The maximal content(Cmax,mg/g),time to peak (Tmax,h),clearance ratio (Ke/h),high-life period (t1/2) and area under the concentration-time curve(AUC0-∞,mg/(g · h)) of GB in various time points in retina after a single oral dose were calculated and compared between two groups.Results The standard curve was obtained over the concentration range of 1-32 mg/L with a linear regression equation,Y =0.0732X + 0.056 (r =0.992).A similar content-time curve was seen between GB suspension group and GB-SMEDDS group.The GB content was higher in GB-SMEDDS group than that in GB suspension group from 30 minutes through 12 hours after administration of drugs.The Cmax of GB-SMEDDS group and GB suspension group were(15.83±1.84) mg/g and(2.65±0.10) mg/g,the AUC0-∞ were(15.30±0.11)mg/(g· h)and(6.42±0.19)mg/(g · h).Conclusions HPLC-ESI-MS is proved to be a rapid,accurate,sensitive and suitable method for pharmocokinetic study of GB.SMEDDS can raise the concent of GB in retina,and it probably improve the bioavailability of GB.

Adult ; Carcinoma ; complications ; Female ; Humans ; Kidney Neoplasms ; complications ; Solitary Fibrous Tumors ; complications ; Thyroid Neoplasms ; complications

The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

The link between nonresolving inflammation and cancer is well documented. On the one hand, epidemiologic evidence supports that approximately 25% of all human cancer worldwide is caused by nonresolving inflammation. On the other hand, inflammatory cells are found in the microenvironment of most, if not all, tumors. In the tumor micro-environment, inflammatory cells and molecules influence almost every aspect of cancer. MicroRNAs (miRNAs) participate in the initiation and progression of nonresolving inflammation-related cancer by regulating the key genes and related signaling pathways. Further investigation into the molecular mechanisms by which miRNAs carry out their functions will be of great value in the prevention, early diagnosis, and treatment of tumors.
Chronic Disease ; Humans ; Inflammation ; complications ; genetics ; immunology ; Inflammation Mediators ; immunology ; MicroRNAs ; genetics ; Neoplasms ; etiology ; genetics ; Tumor Microenvironment

OBJECTIVETo observe the effects of Ganoderma lucidum spores on Cytochrome C (Cyt-C) and mitochondrial calcium in the testis of NIDDM rats.

METHODSFifty male Wistar rats were divided randomly into three groups: model, ganoderma and normal control, the first two groups injected with 2% STZ through vena caudalis, and the last one with half-and-half sodium citrate/citrate buffer solution. Two weeks after normal diet, glucose tolerance tests were performed and the rats with abnormal glucose tolerance from the model and ganoderma groups received high-fat and high-carbohydrate food, the ganoderma group given Ganoderma lucidum spores (250mg/[ kg x d] ) in addition, both for 10 weeks. Glucose tolerance tests were repeated 1 day before the end of the experiment and the rats were castrated and relevant indexes measured.

RESULTSThe NIDDM model was successfully constructed. In the model group, the levels of mitochondrial Cyt-C and mitochondrial calcium were significantly lower (P <0. 05) while that of the plasma Cyt-C was significantly higher than in the ganoderma and the control groups.

CONCLUSIONCyt-C and calcium ion are involved in the damage of the testis. Ganoderma lucidum spores can protect the testis of NIDDM rats.

Animals ; Calcium ; metabolism ; Cytochromes c ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Male ; Mitochondria ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Reishi ; Testis ; drug effects ; metabolism

OBJECTIVETo explore the effects of retinol acid (RA) and triiodothyronine (T3) on alleviating the impairment of cognitive function by sleep deprivation (SD).

METHODSMale Wistar rats were divided into 4 groups: control group (C group), sleep deprivation group (SD group), sleep deprivation + RA group (SD + RA group) and sleep deprivation + T3 group (SD + T3 group). Open field test (OFT) was used to observe the nervous behavior of the rats after SD and electrophysiological brain stereotactic method was used to test long-term potentiation (LTP) in dentate gyrus (DG) of the rats. Ng protein expression was determined by Western blot.

RESULTSCompared with the SD group, the number of crossing in OFT, the changes of amplitude of population spike (PS) and the expression of Ng protein in hippocampus were higher significantly in the SD + RA and SD + T3 groups. All of these had not significant difference comparing with the C group.

CONCLUSIONRA and T3 may alleviate the restrain state of neural system after SD by augmenting the expression of Ng protein in hippocampus.

Animals ; Cognition ; drug effects ; Dentate Gyrus ; metabolism ; Long-Term Potentiation ; Male ; Neurogranin ; metabolism ; Rats ; Rats, Wistar ; Sleep Deprivation ; metabolism ; psychology ; Triiodothyronine ; pharmacology ; Vitamin A ; pharmacology

To explore the new approach to prevent graft versus host disease (GVHD) by purging ex vivo T lymphocytes of bone marrow graft through Fas-FasL way, FasL-cDNA was transfected into BALB/c mouse bon e marrow cells by liposome ex vivo. The transfected cells were cultured together with BAC (BALB/c x C57BL/6) mouse bone marrow graft. The mixing bone marrow graft was infused into BALB/c mouse recipients after 60Co-gamma irradiation. The mortality, manifestation and pathologic change of GVHD in recipient mice were observed. The CFU-S and Y chromosome from donor mice were detected. The results showed that compared with control group, the mortality in 60 days of the recipients in the experimental group decreased (20% vs 70%, P < 0.01) and the morbidity of GVHD lowered (40% vs 100%, P < 0.01). The CFU-S counts for all groups were at normal level on 20 days after transplantation. The Y chromosome from donor mice was discovered in 70% bone marrow nucleated cells of recipient mice survived over 2 months in the experimental group. It is concluded that mFasL-cDNA transfected mouse bone marrow cells prevent GVHD after culturing together with bone marrow graft, and accelerate hematopoietic reconstitution in recipient mice.
Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Purging ; Bone Marrow Transplantation ; Fas Ligand Protein ; Female ; Genetic Therapy ; Graft vs Host Disease ; Male ; Membrane Glycoproteins ; genetics ; Mice ; Mice, Inbred BALB C ; Transfection

BACKGROUND: Recently, it is thought that endothelial function is a new independent risk factor of atherosclerotic disease. However, the differences in endothelial function between Tibetan and Han nationality populations have not been fully investigated.OBJ ECTIVE: To investigate the differences in endothelial function between Tibetan and Han nationality population.DESIGN: Controlled analysis.SETTING: Department of Cardiology, General Hospital; Department of Cardiology, Tibet General Hospital of Chinese PLA.PARTICIPANTS: Totally 272 Tibetan male subjects, aged (43±9) years, were enrolled in this study to stand for Tibetan nationality populations. All of them were native residents in Lhasa city. And 580 Qinghai-Tibetan railway constructers with Han nationality, aged (42±11) years, were enrolled in this study to stand for Han nationality populations. All of them were male subjects from Sichuan province and lived in Lhasa city for at least 1 year. All the participants received regular physical examination between February and May 2006 in the General Hospital of Tibet Military Area Command of Chinese PLA. All the subjects lived in the same high-altitude area (the altitude of Lhasa is 3 658 m). Informed consents were obtained from all the participants.METHODS: ①Height, body mass, waist circumference, hip circumference, systolic blood pressure(SBP) and diastolic blood pressure (DBP) were measured. Body mass index (BMI) was measured as body mass/height2. ② Measurement of brachial artery flow-mediated dilation (FMD): All the participants, who were in the fasting state, were examined in supine position following 20-minute rest. The room temperature was about 20 ℃. In the right arm, a sphygmomanometer cuff was positioned 5 cm below the antecubital fossa. A 10-MHz transducer (Vivid 7, GE Corporation, USA) was used to image the right brachial artery. After obtaining the baseline imaging, the blood pressure cuff was inflated 50 mm Hg (1 mm Hg=0.133 kPa) above the participant's SBP to occlude the brachial artery for 4 minutes. The brachial artery was then imaged during cuff inflation and 2 minutes after cuff release. After the cuff was released and reactive hyperaemia occurred, that was, flow in the brachial artery increased to accommodate the dilated resistance vessels in the forearm. In order to ensure the reliability of the data, the cuff placement and image record were performed by two designated performers. Computer-assisted analysis software was used to calculate brachial artery diameters. The absolute and relative changes of brachial artery FMD were automatically calculated out with the attached software of Vivid 7 ultrasonic diagnosis instrument. ③Biochemical study: The biochemical parameters were obtained after an overnight fasting for 12 hours. Venous blood was sampled for the measurement of total cholesterol, triglyceride (TG), high-density lipoprotein cholesterol and low-density lipoprotein cholesterol (LDL-C). ④ Analysis of variance was used to evaluate the measurement data. Chi-square statistic was used to compare enumeration data.MAIN OUTCOME MEASURES: Comparison of change in BMI, waist-hip ratio, blood pressure, blood lipid, baseline brachial diameter and brachial diameter between 2 groups.RESULTS: Totally 272 Tibetan nationality populations and 583 Han nationality populations participated in the final analysis. ① Brachial artery FMD: The baseline brachial artery diameter of Tibetan nationality populations was significantly larger than that of Han nationality population [(4.28±0.06) mm vs. (4.03±0.04) mm, t =71.915 6, P ＜0.01]; The absolute and relative changes of brachial artery of Tibetan nationality populations were significantly smaller than those of Han nationality populations, respectively [(0.124±0.005) mm vs. (0.141±0.006) mm; (2.934±0.204)% vs.(3.587±0.152)%, t = 40.582 0,52.173 2, P ＜ 0.01]. ②Physical study results: BMI and waist-hip ratio of Tibetan nationality populations were significantly larger than those of Han nationality populations [(30.1±2.5) kg/m2 vs. (26.5±3.4) kg/m2, 0.92±0.07 vs. 0.88±0.05, t =15.595 1, 9.525 4, P ＜ 0.01]. ③TG and LDL-C levels of Tibetan nationality population were (2.31±1.31) mmol/L and (3.49±0.91) mmol/L, respectively, which were significantly higler than those of Han nationality population [(1.97±1.44) mmol/L and (3.07±0.86) mmol/L, t =3.420 0, 6.522 3, P ＜ 0.01].CONCLUSION: ① Brachial artery FMD of Tibetan nationality population is poorer than that of Han nationality population,I.e. Poor vascular reactivity. ② Tibetan nationality populations have severe abdominal obesity and higher level of blood lipid as compared with Han nationality populations.

DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.
China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Pinus ; classification ; genetics ; Pollen ; classification ; genetics ; Quality Control ; Typhaceae ; classification ; genetics