Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P＜0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.

OBJECTIVETo dynamically assess drug targeting of Yougui Pill (YP) and Zuogui Pill (ZP) using infrared thermography.

METHODSIn this self-control experiment, five healthy volunteers were recruited. By using infrared thermography 10 to 11 thermal images of different body locations were taken from each participant after they took warm water, YP, ZP, and their dissembled prescriptions at 30, 70, 100, 130, and 160 min, respectively. The heat values in the lower quadrant abdomen, uterus, Du channel, and Shenque (CV8) were statistically analyzed after scanning for 125 times.

RESULTSAdministration of YP and its disassembled prescriptions enhanced the heat value of the locations of the Du channel and Shenque (CV8), but did no enhance the heat value of the lower quadrant abdomen at 30 min. Administration of ZP and its disassembled prescriptions reduced the heat value in the locations of the lower quadrant abdomen, uterus, Du channel, and Shenque (CV8) at each time point.

CONCLUSIONThe drug targeting of ZP and YP focused on the locations of the Du channel and Shenque (CV8), not on the locations of the lower quadrant abdomen or uterus.

Adult ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Infrared Rays ; Thermography ; methods ; Young Adult

Adolescent ; Adult ; Athletic Injuries ; Clinical Clerkship ; Female ; Humans ; Nurses ; Occupational Injuries ; Young Adult

Background The retinal degeneration 11 (rd11) mouse is a newly discovered naturally occurring recessive animal model with lysophosphatidylcholine acyltransferase 1 (Lpcatl) mutation.Previous studies showed that the photoreceptor cells are characterized by typical rod-cone degeneration pattern in rd1 1 mice,while cone degeneration pattern in rd11 mice is unclcar.Objective Using immunofluorescence staining techniques with retinal wholemount,we aim to clarify the degeneration patterns of cone-function related M-opsin or S-opsin in different ages of rd1 1 mice.Methods A total of thirty rd1 1 and C57BL/6J mice at postnatal (P) day 14,28,42 (five in each age group) were sacrificed and retinal wholemounts were prepared.Immunohistochemistry was performed to identify the expression of M-opsin or S-opsin in retinal wholemounts,which were photographed with a fluorescent microscope.Cone opsins were compared between rd1 1 retinas and age-matched normal C57BL/6J retinas by manually counting the opsin positive cone cells in different quadrants of the retinas.Results The number of M-opsin or S-opsin positive fluorescent dots in each quadrant was similar at all ages of normal C57BL/6J retina.M-opsin positive fluorescent dots in dorsal/temporal,ventral/temporal,dorsal/nasal and ventral/nasal quadrants of rdl 1 retina at P28 were (414±32),(300± 8),(324 ± 22) and (250± 20)/0.037 mm2,which were lower than the age-matched normal C57BL/6J mice (t =4.114,15.225,7.505,17.990,all at P＜0.05).At the same time the S-opsin positive fluorescent dots in P28 rd11 were (8 ±4),(175 ± 16),(74 ± 13) and (315 ±20)/0.037 mm2,with significant decrease in comparison with those in the age-matched normal C57BL/6J mice (t =8.555,17.076,21.637,13.498,all at P＜0.05).With the development of retinal degeneration in rd11 mice,the M-opsin degeneration spread from central to ventral,nasal and then to temporal and dorsal peripheral retina;and the S-opsin loss started from dorsal/temporal to ventral/nasal retina.Conclusions Most of the M-opsin and S-opsins,especially the S-opsins in rd11 mice,degenerate in 6 weeks.Retinal wholemount and cone opsin immunofluorescent staining provide a useful tool to show the cone degeneration pattern and to evaluate the therapeutic efficiency in ongoing gene therapy study.

Background Trans-corneally subretinal injection in rodent model is a useful method for genetic therapy,stem cell transplantation and the study on the ophthalmic research.Standarized operation process is critical for the successful treatment.However,there is no literature to report the detailed procedure and the influence of this technique on morphology and function of retina.Objective This sudy was to introduce a method of trans-corneally subretinal injection and evaluate its influence on the morphology and function of retina.Methods Trans-corneallly subretinal injection was performed on the left eyes of 2-month-old SPF C57BL/6J mice after dilation of pupils.A 301/2G disposable needle was used to puncture the cornea within the pupil area near limbus and avoid touching the lens and irises under eye surgery microscope.Then,a 33G blunt needle was used to insert into the vitreous and toward subretinal space via corneal puncture.Normal saline with 0.1％ fluorescein sodium of 1 μl was slowly injected into the space,and 2.5％ hydroxypropyl methylcellulose was dropped on ocular surface for the observation of the fundus clearly.According to the percentage of the retina filled with subretinally injected solution,the experimental eyes were divided into 80％-100％ area group,50％-70％ area group after injection,and the mice in the pseudo-injected group,in which injection procedure stopped just before the solution was pushed in to the subretinal space did not inject any solution after punctured.The right uninjected eyes of the mice served as normal control group.Four eyes were selected for each group.The structural changes were evaluated by optical coherance tomograpby (OCT) 1 day,2 days,3 days and 5 weeks after injection,and retinal function was assessed by the recored of electroretinography (ERG) 5 weeks after injection.The retinal sepcimens were prepared to examin the morphological changes by hematoxylin and esosin staning.The use of care followd the Regulations for the Administration of Affair Concerning Experimental Animals of Zhejiang Province.Results About 70％ of the injected eyes showed that retinal blebs filled with injected green fluorescein solution occupied 50％ or more retinal area with minimal damages.The focal detachment between neurosensory retinal layer and retinal pigment epithelium (RPE) was exhibited 1 day postinjection,and almost all the retinas retached 2 days after injection.In the fifth week after injection,the amplitudes of ERG b wave were (386.25±37.88),(357.50±41.03),(324.25±53.45) and (410.50±14.88) μV in the sham operation group,50％-70％ area group,80％-100％ area group and normal control group,respectively,showing a significant difference among the 4 groups (F=3.574,P=0.047),and the amplitudes of b wave in the normal control group were higher than those in the 80％-100％ area group (all at P ＜ 0.05).The detachment between retinal neuroepithelium layer and RPE layer,cell proliferation and transposition in the outer nuclear layer were dispalyed under the light microscope in the sham operation group,50％-70％ area group and 80％-100％ area group,and the disordered outer segment of photoreceptors at the injecting area was seen in the 50％-70％ area and 80％-100％ area groups at five weeks after injection.However,retinal sructure and morphology were normal at the non-injection area.Conclusions Trans-corneally subretinal injection is an effective and safe way for subretinal injection.

OBJECTIVETo determined hydrogen selenide in workplace air with atomic fluorescence.

METHODHydrogen selenide were sampled with 0.1 mol/L sodium hydroxide solution in multi-hole absorbing tubes. The sampled absorbing solution were digested with (9+1) nitric acid/perchloric acid. The selenide in sample were reduced by potassium borohydride in 5.0% hydrochloride solution and determined with atomic fluorescence.

RESULTSThere was a good linearity (r=0.9999) over the concentration of 0-150 microg/L, The precision of low, middle and high concentration were 3.1%, 7.4% and 6.7%, respectively. The sample collection rate can reach 99%.

CONCLUSIONThe method was accurate and sensitive to detect hydrogen selenide in workplace air.

Air Pollutants, Occupational ; analysis ; Selenium Compounds ; analysis ; Spectrometry, Fluorescence ; Spectrophotometry, Atomic ; Workplace

Objective To investigate the effects of mesenchymal stem cells (MSCs) transplantation combining with vascular endothelial growth factor (VEGF) gene therapy on myocardium rebuilding,angiogene- sis,and heart function improvement in rats with myocardial infarction.Methods SD rat MSCs were isola- ted,cultured in vitro,labeled with BrdU and transfected by Ad.VEGF gene.Four weeks after left anterior descending artery was ligated to created rat myocardial infarction,cardiac function was examined with echocar- diography,rats were randomly divided into four groups (n=10 in each group):GroupⅠ:MSCs/Ad.VEGF implantation;GroupⅡ:MSCs implantation;GroupⅢ:Ad.VEGF injection;GroupⅣ:Control.MSCs dif- ferentiation was observed 4 weeks after transplantation.Immunohistochemistry and angiogenesis were observed. Echocardiography was performed to detect the effects on heart function.Results MSCs labeled with BrdU could be identified in host hearts in groupⅠandⅡ,most of them positively stained with cTnT antibody. Echocardiography indicated that the improvement of the LVEF value in groupⅠwas more significant than that in the other three groups (P＜0.01,respectively).Some cells were incorporated into the coronary capillaries in the infarcted region.The capillary density in groupⅠwas higher than that in the other three groups (P＜0.01,respectively).Conclusion MSCs implantation combining with VEGF gene therapy can obviously re- pair damaged myocardium and enhance the angiogenesis in ischemic heart tissue.

The commodity specification and grade of Chinese medicinal materials is a measure of the quality of traditional Chinese medicines (TCMs), which directly impacts on the safety and effectiveness of clinical medicines. It is an urgent problem to establish a set of standards which can both interpret the scientific connotation of the commodity specification and grade of Chinese medicinal materials and play a significant role on clinical medicines as well as markets. This paper reviews the research methods of the commodity specification and grade of Chinese medicinal materials such as sensory evaluation, chemical assessment, biological evaluation, and cited the applications of various methods for the classification of TCMs. It provides technical support for establishing standards of the commodity specification and grade of Chinese medicinal materials, and also constructs scientific basis for clinical rational drug use.
Drugs, Chinese Herbal ; chemistry ; economics ; Humans ; Medicine, Chinese Traditional ; economics ; standards ; Plants, Medicinal ; chemistry ; Quality Control ; Research Design

Estrogen receptor alpha (ERalpha) has been a primary target of treatment as well as a prognostic indicator for breast cancer. The level of human X-box binding protein 1 (XBP-1) mRNA was related with that of ERalpha in breast tumors and was over-expressed in some breast tumors. These previous studies suggested that XBP-1 may interact with ERalpha. XBP-1 has two isoforms, XBP-1S and XBP-1U, as the result of unique splicing. GST pull-down assay showed that both XBP-1S and XBP-1U bound to ERalpha in vitro. The binding of XBP-1S to ERalpha was stronger than that of XBP-1U to ERalpha. Co-immunoprecipitation revealed that the binding was in a ligand-independent manner. XBP-1S and XBP-1U interacted with the region of ERalpha that contains a DNA-binding domain. The ERalpha-interacting regions on XBP-1S and XBP-1U have been mapped to two regions, the N-terminal basic region leucine zipper domain (bzip) and the C-terminal activation domain. These findings suggest that XBP-1S and XBP-1U may participate in ERalpha signaling pathway through the mediation of ERalpha.
Breast Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; DNA-Binding Proteins ; genetics ; metabolism ; Estrogen Receptor alpha ; genetics ; metabolism ; Female ; Humans ; Protein Interaction Domains and Motifs ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Regulatory Factor X Transcription Factors ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; X-Box Binding Protein 1

OBJECTIVETo construct a prokaryotic expression system to serially express Alloferon-1 and to determine the anti-tumor activity of its products in vitro.

METHODSAn artificial fusion gene containing 6 x His-EK-8 x Alloferon-1-EK-6 x His sequences was constructed by linking primer PCR. By using routine molecular biological methods, the artificial fusion gene was cloned and its prokaryotic expression system was then constructed. SDS-PAGE and BioRad Agarose Image Analysor was applied to measure the expression and output of the target recombinant products 8 x rAlloferon-1-EK. Ni-NTA affinity chromatography and EK digestion and Sephadex G-50 chromatography were performed to extract 8 x rAlloferon-1-EK and rAlloferon-1-EK, respectively. The proliferation of KB, SGC and HL-60 tumor cells was tested by using MTT method after treatment with directly synthesized Alloferon-1 (sAlloferon-1), Aloferon-1-EK (sAlloferon-1-EK) and rAlloferon-1-EK.

RESULTThe target artificial fusion gene and its prokaryotic expression system pET42a-8 x rAlloferon-1-EK-E. coliBLDE3 with the expected sequences were obtained. Under inducement of IPTG, the prokaryotic expression system expressed the target serial recombinant protein 8 x rAlloferon-1-EK and its output was approximate 30 % of the total bacterial proteins. 8 x rAlloferon-1-EK and rAlloferon-1-EK were obtained through Ni-NTA and Sephadex G-50 columns. sAlloferon-1, sAlloferon-1-EK and rAlloferon-1ìrAlloferon-EK showed similar remarkable effects of inhibiting the growth and proliferation of KB, SGC and HL-60 cells in vitro within 25 approximately 100 microg/ml concentration range (P<0.01), and there were no significant differences in the inhibiting effects among the three agents (P>0.05).

CONCLUSIONA prokaryotic expression system to serially express rAlloferon-1 has been successfully constructed. The product rAlloferon-1-EK has a similar anti-tumor activity compared to both the synthesized Alloferon-1 and Alloferon-1-EK in vitro.

Animals ; Anti-Bacterial Agents ; pharmacology ; Antineoplastic Agents ; metabolism ; pharmacology ; Bacterial Vaccines ; administration & dosage ; genetics ; metabolism ; Cell Line, Tumor ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Growth Inhibitors ; genetics ; metabolism ; pharmacology ; HL-60 Cells ; Helicobacter pylori ; genetics ; metabolism ; Humans ; KB Cells ; Peptides ; genetics ; immunology ; metabolism ; pharmacology ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology