Objective To investigate the changes of expression of neuronal nitric oxide synthase(nNOS) after sleep deprivation(SD) in rats,and to study the effects of Xingnaojing on them. Methods Rapid eye movement(REM) SD models(SD group,n=40) were established by the modified multiple platform methods.The rats in SD group were subjected to drug intervention group(n=20) and non-drug intervention group(n=20).Xingnaojing injection was administered to the drug intervention group.Rats in SD group were deprived of sleep for 1,3,5 and 7 d. The expression of nNOS in brain tissues was detected by immunohistochemistry.The rats without SD were severed as cage control group(n=10) and tank control group(n=10). Results The expression of positive cells of nNOS in brain tissues increased with the time of SD,and Xingnaojing decreased the number of positive cells of nNOS.The area density of positive reaction cells in SD group was significantly higher than those in cage control group and tank control group(P

Objective To explore the metabolic characteristic of prostate cancer (PCa) in central gland with magnetic resonance (MR) spectroscopic, and evaluate the value of MRS in the differential diagnosis of benign prostatic hyperplasia nodus. Methods MR images were performed in 38 cases with prostate disease by 3.0T MR, 11 cases with PCa in central gland and 27 cases with benign prostatic hyperplasia nodus. All the cases were scanned by routine, then by the combined MRS. MRS findings were reviewed in 27 cases with benign prostatic hyperplasia (BPH) and 11 cases with PCa (3 in central gland origin, 8 with large tumor invading both peripheral zone and central gland). (Cho+Cr)/Cit ratios of PCa and BPH were retrospectively measured, (Cho+Cr)/Cit of PCa voxels were compared with that of BPH voxels. Results Significantly higher choline levels and lower citrate levels were observed in central gland of PCa compared with BPH. In the glandular BPH region, the amount of Cit was high; while in the stromal region, the Cit and Cho level was much lower. The average (Cho+Cre)/Cit values of PCa and BHP were 1.94±1.43 and 0. 83±0. 28 respectively, the difference in ratio between both was statistically significant (P＜0. 01). Conclusions The combined use of MRI and MR spectroscopy is propitious for differentiating prostate cancer in central gland and benign prostatic hyperplasia nodus and for increasing the diagnostic accuracy of prostate cancer.

Objective To evaluate the clinical outcome of a method of renconstruction of Type Ⅱb defect of thumb both and reserving the length of the donor toe.Methods From March 2012 to Febrary 2014,there were 11 cases of thumb defect treated in our hospital.According to Gu Yudong's classification,all were type Ⅱ b.Three were open defect and others were closed defect.All were treated with combined transfer of big toe wrap-around flap and autogenous iliac graft for thumb reconstruction while a superficial circumflex iliac artery perforator flap was used to resurface the donor defect.The donor site of the belly was sutured directly.Results All of the reconstructed thumb survived.Among 11 flaps,arterial crisis occurrred in 1 case,venous crisis occurred in 2 cases,and all survived after operative treatment.The patients were followed-up from 3 mooths to 15 months,all the thumbs had a good appearance.The thumb opposition function was good and sensation recovery to S3 + and two-point discrimination from 6-8 mm.The donor big-toe was preserved.All the flaps had satisfactory color and texture,but looked a little plump.All the patients had no effect on walking and running.Four flaps had more beautiful appearance after flap plastic.There was only one linear scar on the belly donor without any discomfort.Conclusion It is an effective method of combined transfer of big toe wrap-around flap and autogenous iliac graft while a superficial circumflex iliac artery perforator flap was used to resurface the donor defect used to reconstrucution of Type Ⅱ b defect of thumb is very good.

Objective To explore the changes of intereferon-?(IFN-?)and interleukin-4(IL-4)in peripheral blood of children with mycoplasma pneumoniae(MP) encephalitis at the acute phase.Methods The peripheral blood concentrations of IFN-? and IL-4 were measured by enzyme-linked immunosorbent assay(ELISA) in 24 cases of children with MP encephalitis at the acute stage.The samples from 24 cases of healthy children were control group.Twenty-four children with MP encephalitis were intravenous drip with azithromycin,at the same time,10 cases received hexadecadrol and 15 cases received gamma globulin.Results The serum concentrations of IFN-? and IL-4 in the mycoplasmal encephalitis group were(98.56?12.93) and(45.55?17.58) ng/L,respectively,which were significantly higher than those in control group [(85.35?6.91) and(26.78?9.89) ng/L] respectively(Pa

Objective To explore the methods and curative effect of metallic self-expanding stent in inoperable malignant gas-troduodenal obstruction. Methods The data of 15 cases with gastroduodenal obstruction including 9 cases of carcinoma of head of pancreas and 6 cases of carcinoma of stomach were analyzed retrospectively. The operative procedures of the stent implanted and the tors accepted more radiation dose because the manipulation was under the fluoroscopy in a short distance and with a full field of view. sions, the postoperative eating habit and the development turnover of disease. The main death reasons were tumor transfer and sys-tem exhaustion. Conclusion To pay close attention to the details and main points of operative procedure is the key point to implant stent successfully for malignant gastroduodenal obstruction. The determinative factor to influence the curative effect is the develop-ment turnover of tumor.

Objective To identify a unique protein as a novel genetic marker for rapid molecular typing of Mycobacteriutn tuberculosis Beijing genotype strains by comparing the proteome of Beijing genotype strains with non-Beijing strains.Methods Fifty-six clinical isolates of Mycobacterium tuber- culosis were analyzed by spoligotyping to determine genotypes.The two-dimensional electrophoresis (2 DE)was used to compare the global protein patterns between Beijing genotype strains and non Bei jing strains.Differential expressed proteins were measured by matrix assisted laser desorption ioniza tion lime of flight mass spectrometry(MALDI-TOF-MS).The data obtained from peptide mass fingerprinting were compared in protein database.The genes encoding differential expressed proteins and their upstream were sequenced.Results Forty nine of the 56 isolates were Beijing genotype strains and 7 isolates were non-Beijing strains.A unique protein Rv0927c was identified,which is absent in Beijing genotype strains compared with 7 non Beijing strains and H37Rv.There were two characteristic mutations in Beijing genotype strains,a deletion of AGC at nucleotide position 421 of Rv0927c and a 127 G→A muta- tion in the upstream of Rv0927c.but not in non Beijing strains and H37Rv.Conclusion Characteris tic mutations of Rv0927c in Beijing genotype strains can be used as a novel genetic marker for rapid molecular typing of Mycobacteriuln tuberculosis Beijing genotype strains and non Beijing strains.

BACKGROUNDCardiomyocyte transplantation for the therapy of myocardial ischaemia is being paid close attention. However, how the microenvironment controls the differentiation of transplanted bone marrow stromal cells (BMSCs) is unknown. Endothelin-1 (ET-1), a cytokine, increases during myocardial infarction, but it is not known whether ET-1 is responsible for the fate of transplanted BMSCs. In the present study, we investigated the effects of ET-1 on differentiation and maturation of induced rabbit BMSCs, in vitro, to elucidate the cellular biological mechanisms.

METHODSThe proliferation of BMSCs isolated from femur of rabbits was induced by ET-1 only, by 5-azacytidine (5-aza) or ET-1 combined with 5-aza. After 4 weeks of induced culturing, the differentiation rate and the diameter of induced myocyte like cells were estimated and the expressions of GATA-4 protein and phosphorylation level were assayed by Western-blot, RT-PCR analysis of beta-myosin heavy chain (MHC). mRNA expression, levels of troponin-I by immunohistochemical staining and ultrastructure of induce-cultured BMSCs were also determined.

RESULTSBy induction with ET-1 and 5-aza, mean cell diameter of induced BMSCs was larger than induced with 5-aza [(6.26 +/- 0.22) microm cf (5.29 +/- 0.19) microm] (P < 0.001). There was no difference in rate of differentiation of myocyte like cells between the groups induced with 5-aza and ET-1 combined with 5-aza [(29.82 +/- 0.23)% cf (29.94 +/- 0.18)%] (P > 0.05). The expressions of GATA-4 protein and phosphorylation were enhanced significantly in groups induced with ET-1 combined with 5-aza (P < 0.05). In the group induced with ET-1 combined with 5-aza, expression of beta-MHC mRNA was higher than control [(0.122 +/- 0.008) cf (0.022 +/- 0.003)] (P < 0.01), and more troponin-I positive cells were also detected in this group. Differentiated BMSCs showed formations of myofilaments and primitive sarcomere, i.e., morphological characteristics of myocyte like cells.

CONCLUSIONSThis study suggests that induced culturing of BMSCs by ET-1 combined with 5-aza can express cardiomyocytic characteristics whereas ET-1 alone could not induce BMSCs to differentiate to myocyte like cells. ET-1 upregulates the expression of GATA-4 protein and phosphorylation level of induced BMSCs, and rapidly promotes the differentiation and maturation of myocyte like cells from BMSCs.

Animals ; Bone Marrow Cells ; cytology ; ultrastructure ; Cell Differentiation ; drug effects ; Cells, Cultured ; Endothelin-1 ; pharmacology ; GATA4 Transcription Factor ; analysis ; metabolism ; Myocytes, Cardiac ; cytology ; Myosin Heavy Chains ; genetics ; Phosphorylation ; RNA, Messenger ; analysis ; Rabbits ; Stromal Cells ; cytology ; ultrastructure

Background Human limbal allograft transplantation or limbal autograft transplantation are the primary approaches to the severe corneal-blindness,but their application in clinic were limited because of the defects of donor material.With the development of tissue engineering technology,transplantation of in vitro cultured limbal epithelial stem cells is being an advanced management.Objective The aim of this work was to expand human limbal epithelial stem cells ex vivo under the guidance of confocal microscope and to lay the foundation for fabricating ex vivo cultured cell sheets.Methods Ten eyes of ten patients were examined with the Heidelberg Retina Tomography Ⅲ Rostock Cornea Module(HRT3-RCM)to elucidate the structure of the human corneoscleral limbus and to correlate limbal epithelial dimensions.According to the analysis of the images of limbal epithelia,the limbal tissues provided by Eye Bank of Henan Eye Institute were cut into suitable explants.Then,this study was conducted to expand limbal epithelial stem cells ex vivo on denuded amniotic membrane.The phenotypes of primary cultured cells were evaluated by morphology and immunofluorescent staining with antibodies for limbal epithelial stem cell markers (p63,cytokeratinl9)and differentiation markers(keratin 3,involucrin).This experimental procedure was approved by the Ethic Committee of Henan Provincial People's Hospital.The written informed consent was obtained from subjects before initiation of any examination.Results The palisade morphology of human limbus was imaged clearly on the laser scanning in vivo confocal microscopy and many hyperreflective cells were observed in palisade basal cells.The cell-island phenomenon was seen in the basement membrane under the laser scanning in vivo confocal microscopy.The oblique sections of limbus showed many papilla-like epithelial columns below the superficial limbal epithelia.Throughout the experiment duration,the epithelial cells grew well with the migration rates from limbal tissue (68.62± 16.94)％ and the migration time(5.83 ±2.04)days,which depended on the tissue freshness.Compared with the second and forth batch of tissue,the migration rates of the third and sixth batch of tissues were significantly higher(P＜0.05),and the migration time was evidently longer in the forth and sixth batch of tissue compared with the first,second,third and fifth batch(P＜0.05).The positively expressing rates in the cultured corneal stem cells were 4.05％ and 36.52％ for p63,26.07％ and 40.55％ for CK19,57.88％ and 40.81％ for K3,64.66％ and 59.19％ for involucrin.Conclusion Human limbal epithelial stem cells can be successfully and purposefully obtained from the limbal tissue based on the guidance confocal miscroscope.The cultured corneal stem cells can grow well on the denuded amniotic membrane

OBJECTIVETo establish a new method for simultaneous determination of shanzhiside methyl ester, chlorogenic acid, 8-O-acetyl shanzhiside methylester, forsythiaside B, rutin, acteoside and galuteolin in Lamiophlomis rotata.

METHODSeparation was performed on a Welchrom-C18 chromatographic column with acetonitrile-0.1% orthophosphoric acid as mobile phasewith gradient elution. The flow rate was 1.0 mL x min(-1). The column temperature was 30 degrees C, and the detection wavelength was set at 238 nm, 330 nm and 350 nm.

RESULTThe seven compounds were well separated with good linear correlations. The mean recoveries of seven compounds were 96.47%-102.2% (RSD 0.70%-2.2%).

CONCLUSIONThere were good correlations among the seven compounds in the samples of aerial parts. The mean sum of shanzhiside methyl ester and 8-O-acetyl shanzhiside methylester in samples of aerial parts is 1.44%. The aerial parts have more kinds of composition and with higher content than that of underground parts in L. rotata, which was reasonable for the resonable use of the aerial part as medicinal part. The method was simple, repeatable and stable, which could be used for identification and quality evaluation of L. rotata.

Chlorogenic Acid ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Glycosides ; chemistry ; Lamiaceae ; chemistry ; Methyl Ethers ; chemistry ; Phenols ; chemistry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Pyrans ; chemistry ; Rutin ; chemistry

Antigens, CD20 ; metabolism ; CD5 Antigens ; metabolism ; Colonic Neoplasms ; complications ; metabolism ; pathology ; surgery ; Cyclin D1 ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Ileal Diseases ; complications ; pathology ; surgery ; Ileocecal Valve ; Intestinal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Intestinal Polyps ; complications ; metabolism ; pathology ; surgery ; Intussusception ; complications ; pathology ; surgery ; Leukemia, Lymphocytic, Chronic, B-Cell ; metabolism ; pathology ; Lymphoma, Mantle-Cell ; complications ; metabolism ; pathology ; surgery ; Middle Aged