OBJECTIVETo observe the clinical significance of nitric oxide (NO) and 8-isoprostane (8-isoPG) changes in exhaled breath condensate ( EBC) of acute respiratory distress syndrome (ARDS) patients after treated by Qingfei Decoction (QD).

METHODSTotally 48 ARDS patients receiving mechanical ventilation were equally assigned to the QD treatment group and the control group by random digit table. EBC specimens were collected by modified Ecoscreen breath condensate collector (German JAEGER Company) on the first day and the fifth day after confirmed diagnosis of ARDS. Concentrations of NO and 8-isoPG in EBC were measured by ELISA. The oxygenation index and APACHE II scores were recorded at the same time.

RESULTS(1) The fatality rate in the QD treatment group was lower than that in the control group (8.3% vs 37.5%, P < 0.05). (2) After treatment NO and 8-isoPG concentrations in EBC were lower in the QD treatment group (34.49 ± 5.67 µmol/L, 30.09 ± 7.89 ng/L) than in the control group (39.78 ± 9.27 µmol/L, 35.65 ± 8.90 ng/L; P < 0.05). (3) After treatment improved oxygenation index value was higher in the QD treatment group than in the control group (120.88 ± 35.16 vs 101.50 ± 37.70, P < 0.05). After treatment APACHEII scores was lower in the QD treatment group than in the control group (6.21 ± 3.51 vs 10. 26 ± 4.33, P < 0.05).

CONCLUSIONTreatment of ARDS patients by QD was favorable in controlling inflammation, alleviating lung injury, and improving clinical efficacy.

Breath Tests ; Dinoprost ; analogs & derivatives ; analysis ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Inflammation ; Nitric Oxide ; analysis ; Respiration, Artificial ; Respiratory Distress Syndrome, Adult ; drug therapy

To detect the in vitro probe microdialysis recoveries based on an HPLC-DAD method for simultaneous quantification of nine active ingredients (ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and glycyrrhizic acid) in Mahuang decoction, which provides reference for in vivo pharmacokinetic study. The concentrations of nine active ingredients in dialysate were detected by HPLC-DAD, to investigate the effect of flow rates (incremental method and subtraction method) and intraday stability of the probe recoveries and medium concentrations on the recoveries. Nine active ingredients could be well separated in 52 min. At the perfusion rate of 1.0 μL x min(-1), the relative recoveries of ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamyl alcohol, cinnamic acid, cinnamaldehyde and glycyrrhizic acid were (50.95 ± 0.82)%, (52.74 ± 1.13)%, (51.29 ± 0.51)%, (32.56 ± 0.84)%, (45.36 ± 0.83)%, (70.94 ± 0.99)%, (69.98 ± 2.30)%, (71.68 ± 0.63)%, and (22.14 ± 0.48)%, respectively. And the probe kept steady in 7 hours. At the same medium concentration, the probe recoveries decreased exponentially with the increase in flow rates. The recoveries of seven ingredients detected by these two methods were similar at certain flow rates, except for amygdalin and cinnamaldehyde. At the same flow rate, the relative recoveries of cinnamyl alcohol, cinnamic acid and cinnamaldehyde changed greatly (9.55%-16.2%) and the others six ingredients had less change (3.27%-5.71%) with the changes in medium concentrations. Microdialysis method could be used to detect the in vitro recoveries of nine ingredients in Mahuang decoction. Reverse dialysis method could be used for the in vivo probe recovery calibration of ephedrine, pseudoephedrine, methylephedrine, liquiritin, cinnamyl alcohol and cinnamic acid at the flow rate of 2.0 μL x min(-1).
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ephedra sinica ; chemistry ; Microdialysis ; methods ; Molecular Structure

BackgroundHuman retinal gliocytes play an important role in proliferative diseases,which are the basis of in vitro studies.Researchers have cultured human retinal gliocytes in the past.In our study,we found that the cells we cultured presented a unique shape different from those by other researchers.ObjectiveThis study was to design to produce a new culture and purification method for retinal gliocyte in vitro.Methods Retinal tissue was isolated from human eyeballs and digested using the two-step digestion method (2％ pancreatin and 0.133％collagenase Ⅵ) to harvest the retinal glio cytes.The cells were collected and cultured in endothelial cell-targeted nutrient culture containing 10％ fetal calf serum and supplemented with β-endothelial cell growth factor (ECGF) and sodium heparin,and the culture dishes were coated with fibronectin(FN) to promote the attachment of retinal gliocyte.During the culturing process,the gliocytes were identified by the observation of morphological characteristic and regular histological examination.The identification of the cells also was performed by immunochemistry targeting glial fibrillary acidic protein (GFAP),Vimentin,neuron specific enolase ( NSE ),S-100,CD34,and Ⅷ factor.Results Retinal gliocytes were isolated successfully from the human retina by the two-step digestion method.Primary cultured cells attached after 72 hours and achieved confluency between day 9 and 10 that were aligned petaliform in shape.Regular histological examination after H&E staining showed blue cell nuclei and light red cytoplasm.The target cells presented with strong responses for GFAP and Vimentin and no response for NSE,S-100,CD34 and Ⅷ factor.ConclusionsLarge amount of purified human retinal gliocytes can be obtained by two-step digestion and cultured in endothelial cells-targeted culture medium supplemented with β-ECGF and sodium heparin in plates coated with FN.The cultured cells expressed markers for retinal gliocytes.However,specific features of these cells remain to be further elucidated.

OBJECTIVETo explore the feasibility and practical value of fluorescence in situ hybridization (FISH) detection of TERC gene amplification in cervical intraepithelial lesions (CIN) and squamous cell carcinoma (SCC).

METHODSTissue microarray was constructed to cover 150 cases of various cervical conditions, including 24 cases of normal cervical mucosa, 78 cases of CINs (CINI, 25 cases; CINII, 21 cases and CINIII, 32 cases) and 48 cases of SCC. FISH was used to detect TERC gene amplification.

RESULTSTERC gene amplification was detected in 8% (2/25) CINI, 47.6% (10/21) CINII, 71.9% (23/32) CINIII and 87.5% (42/48) SCC. There were significant differences among these groups (P < 0.05). The amplification rates of TERC gene in SCC, CINIII and CINII were significantly higher than those of normal cervical epithelium and CINI (P < 0.05). Significant differences were also observed among CINI and CINII, CINIII and SCC (P < 0.05), and between CINII and SCC (P < 0.05). There were no significant differences between normal cervical epithelium and CINI, CINII and CIN III, and between CINIII and SCC (P > 0.05). FISH detection of amplification of TERC gene in CINI and CINII-III demonstrated the following statistics: sensitivity of 62.3%, specificity of 92.0%, accuracy of 71.8%, positive and negative predictive values of 94.3% and 53.5%, respectively.

CONCLUSIONSFISH detection is a reliable method in detecting TERC gene amplification using paraffin tissue sections. When histological evaluation becomes difficult, TERC amplification detectable by FISH may offer a diagnostic distinction of CINI from CINII. Moreover, TERC amplification may be used as a biomarker in predicting CIN progression to invasive cancer.

Adenoma ; diagnosis ; genetics ; Adult ; Aged ; Biomarkers, Tumor ; analysis ; Carcinoma, Squamous Cell ; diagnosis ; genetics ; Cervical Intraepithelial Neoplasia ; diagnosis ; genetics ; Disease Progression ; Female ; Gene Amplification ; Humans ; In Situ Hybridization, Fluorescence ; Middle Aged ; RNA ; genetics ; Sensitivity and Specificity ; Telomerase ; genetics ; Uterine Cervical Neoplasms ; diagnosis ; genetics ; Young Adult

Objective · To investigate the impacts of hepatitis B virus (HBV) infection on the metabolomic phenotype of HepG2 human hepatoma cells.Methods · With gas chromatography-mass spectrometry (GC-MS), metabolite composition of HepG2 and HepG2.2.15 cells (derived from HepG2 cells transfected with a plasmid containing HBV) were analysed. Results · GC-MS analysis mainly found 34 metabolites in both HepG2 and HepG2.2.15 cells,including glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), serine (Ser), threonine (Thr), methionine (Met), cysteine (Cys), cystine, aspartic acid (Asp), glutamic acid (Glu), pyroglutamic acid, phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), hypoxanthine, uracil,myo-inositol, lactic acid, succinic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, urea, cholesterol, etc. These metabolites were involved in multiple metabolic pathways including glycolysis and metabolism of fatty acids, amino acids, purines and pyrimidines. Compared with HepG2 cells,HepG2.2.15 cells had significantly higher levels in lactic acid, linolenic acid, Ala and Cys, but lower levels in Leu, Ile, Val, Phe, Met, Trp, Pro, Tyr, myoinositol and uracil. Conclusion · HBV infection dysregulates the metabolism of amino acids and fatty acids in hepatocytes. GC-MS analysis provides complimentary information about HBV-induced metabolic changes of host cells.

AIM AND METHODSTo observe the effect of myocardial mitochondrial L-arginine (L-Arg)/nitric oxide (NO) system on mitochondrial Ca2+ transport by using purified rat mitochondria and incubation of them in vitro.

RESULTSCompared with control group, incubation of mitochondria with L-Arg (10(-4) mol/L, NO substrate) or sodium nitroprusside (5 x 10(-7) mol/L, the donor of exogenous NO, SNP) increased significantly mitochondrial NO2- (66% and 89%, P < 0.01), respectively, and decreased the Ca2+ content (40% and 54%, P < 0.01). After L-Arg or SNP treatment, mitochondrial Ca2+ uptake were decreased by 67% and 85%, respectively (P < 0.01), vs control. The rate of mitochondrial Ca2+ release decreased by 11% and 8%, respectively (P < 0.01). When L-NAME (NO synthase inhibitor) was incubated with mitochondria and the L-Arg together, it inhibited the effects of L-Arg, NO2 on the mitochondrial NO2 formation, Ca2+ content descending, and decrease of Ca2+ uptake and release.

CONCLUSIONThe data suggest that myocardial mitochondrial L-Arg /NO systems take part in the regulation of cardiomyocytes Ca2+ transportation.

Animals ; Arginine ; metabolism ; Biological Transport ; Calcium ; metabolism ; Female ; Male ; Mitochondria, Heart ; metabolism ; Myocytes, Cardiac ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo identify the risk factors which will indicate the Pneumocystis carinii (Pc) infection in children with systemic lupus erythematosus (SLE) and investigate the clinical features and to elevate the level to find out the high-risk patients and make early diagnosis and treatment.

METHODThe characteristics, clinical features, laboratory examinations, treatment and prognosis of Pneumocystis carinii pneumonia (PCP) in children with SLE under 18 years of age treated in our hospital between January 2000 and January 2013 were prospectively reviewed. A comparison was made with the 26 cases of SLE children without PCP who were matched for gender, age and course, and a literature review was made.

RESULTS(1) Five cases were enrolled, 3 were male and 2 female. Their age range was 13-17 (14.0 ± 1.6) years. All the children had kidney involvement. The courses were from 3 months to 4.5 years. All patients were receiving daily glucocorticoid therapy and immunosuppressive drugs before the diagnosis of PCP.Four patients were in the inactive phase of SLE (SLEDAI 2-4 points), and the fifth case was in active phase (SLEDAI 8, low complement 2 points, anti-dsDNA antibody positive 2 points, urine-protein 4 points). (2) Besides the clinical manifestations of SLE, most patients had progressive dyspnea, fever and dry cough at onset of PCP. Two children accepted mechanical ventilation because of respiratory failure. The mean duration of the symptoms to diagnosis was 10-30 (17.6 ± 7.8) days. Lactose dehydrogenase (LDH) was elevated more or less, median was (700 ± 263) U/L. Lymphocyte count were (0.3-1.4)×10(9)/L (median 0.5×10(9)/L), and three children had CD4 T lymphocyte count <0.3×10(9)/L. Arterial blood gas analyses showed severe hypoxemia. Chest radiographs showed in all cases diffuse interstitial infiltration. Pc was positive in the sputum. All patients were treated with trimethoprim-sulfamethoxazole and corticosteroids.

CONCLUSIONWhen SLE children are treated with corticosteroids and immunosuppressive drugs, low lymphocyte count is the risk factor for Pc infection.It is essential to monitor lymphocyte count.We should pay more attention to fever, dry cough and hypoxemia. Chest radiologic examination may help diagnose the PCP in SLE children.It may be helpful for SLE children whose CD4T lymphocyte was below 0.3×10(9)/L to take trimethoprim-sulfamethoxazole for PCP prophylaxis.

Adolescent ; Anti-Infective Agents ; adverse effects ; therapeutic use ; Case-Control Studies ; Child ; Female ; Glucocorticoids ; adverse effects ; therapeutic use ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Kidney Diseases ; etiology ; Lung ; pathology ; Lupus Erythematosus, Systemic ; complications ; drug therapy ; Lymphocyte Count ; Male ; Opportunistic Infections ; drug therapy ; epidemiology ; Pneumonia, Pneumocystis ; drug therapy ; epidemiology ; Prognosis ; Retrospective Studies ; Risk Factors ; Trimethoprim, Sulfamethoxazole Drug Combination ; therapeutic use

Background The incidence of retinal vascular diseases increase annually,such as diabetic retinopathy,retinopathy of prematurity and age-related macular degeneration.The key of treatment for these diseases is how to evaluate retinal vascular change effectively and objectively.Retro-orbital injection of fluorescein isothiocyanatedextran (FITC-dextran) is a simple and effective method for observing C57BL/6J mouse retinal vessels.But,whether it is suitable for other mice and rats is seldom reported.Objective This experiment was to assess the feasibility of the observation of retinal vessels by retro-orbital injection of FITC-dextran in different genus of mouse and offer the reference for relevant study.Methods Twelve animals of C57BL/6J mice,Kunming mice,SD rats and Wistar rats were selected,respectively and divided into the experimental group and control group at average.The right eyes of the animals of the experimental group received the retro-orbital injection of 9 ml/kg FITC-dextran,and the right eyes of animals of the control group received PBS solution at the same volume and way.All the animals were sacrificed 10 seconds after injection and both eyes of each animal were obtained for retinal stretched preparation.The retrobulbar tissue and whole-mount retina were viewed under a fluorescence microscope.The use of the animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Retinal blood vessels labeled by FITC-dextran could be observed in both eyes of C57BL/6J mice and Kunming mice to present with a green fluorescence in experimental group under a fluorescence microscope,but no any fluorescence-labeled retinal blood vessel was exhibited in the control mice.The retinal blood vessel could not be observed in all eyes of SD rats and Wistar rats after the injection of FITC-dextran both in the experimental group and the control group under a fluorescence microscope.The surrounding tissues of the right eyes of mice and rats dyed with green fluorescence of FITC-dextran in the experimental group,however,green fluorescence could not be seen in the surrounding tissues of the left eyes of mice and rats.Conclusions Retro-orbital injection of FITC-dextran is a suitable method of observing the retinal vessels of mouse but not rat.

In this study, we observed the levels of adrenomedullin (ADM) and proadrenomedullin N-terminal 20 peptide (PAMP) in myocardium and aorta of spontaneously hypertensive rats (SHRs) in comparison with Wistar-kyoto (WKY) rats. Contents of ADM and PAMP were measured by radioimmunoassay (RIA) in plasma, myocardium and aorta. The amount of Pro-ADM mRNA of myocardium and aorta was determined by competitive quantitative reverse transcription polymerase chain reaction (RT-PCR). In SHRs the amounts of Pro-ADM mRNA of myocardium and aorta were 66.7% (P<0.01) and 73% (P<0.01) higher than those in WKY rat, respectively. In SHRs, the levels of ADM in plasma, myocardium and aorta were 29%, 76.7% and 79% (all P<0.01) higher than those in WKY rats, respectively. The level of PAMP in SHRs was increased by 42.5% in plasma (P<0.01), 47.2% in myocardium (P<0.0.1) and 27.3% in aorta (P<0.05) compared to WKY rats, respectively. In addition, the ratio of ADM content to PAMP content in SHRs group was increased compared with that in WKY group (2.0+/-0.25 vs 1.64+/-0.3 and 2.2+/-0.18 vs 1.56+/-0.28, in myocardium and aorta, respectively, P<0.01). These results suggest that ProADM gene expression is up-regulated and the increase in ADM and PAMP is different in SHRs. The significance of inconsistency of increase in ADM and PAMP in SHRs needs to be further investigated.
Adrenomedullin ; genetics ; metabolism ; Animals ; Aorta ; metabolism ; Female ; Male ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Up-Regulation

OBJECTIVE: To investigate the effect of prostaglandin E1 (PGE1) on the expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) in experimental liver fibrosis rats. METHODS: The liver fibrosis model was established by carbon tetrachloride. Rats were divided into a control group and PGE1-treated group. The pathological changes of the liver tissue from the two groups, the semi-quantitative analysis of hepatitic activity in HE stain sections, the pathological image quantitative analysis of the fibrosis degree, TIMP-1 positive cells, and the content of collagen were synthetically analysed. RESULTS: The mark changes of liver pathology in HE stain sections were that the degree of hepatitic activity in the PGE1-treated group was obviously lower than that in the control group (P < 0.05). The fibrosis degree, TIMP-1 positive cells and the collagenous fibers decreased in the PGE1-treated group (P <0.05). CONCLUSION PGE1 has an anti-hepatofibrosis effect in the experimental rats, the inflammation of liver is light, and the proliferation of collagenous fibers can be restrained, whose mechanism is probably associated with the suppression of TIMP-1 expression caused by PGE1.
Alprostadil ; pharmacology ; Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Female ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics