OBJECTIVE:To establish a method for the content determination of rosin acid in Rheumatoid arthritis tablet. METH-ODS:HPLC was performed on the column of ZORBAX SB-C18 with mobile phase of acetonitrile-0.1% formic acid(82∶18,V/V) at flow rate of 1.0 ml/min ,detection wavelength was 241 nm ,column temperature was 30 ℃ and volume injection was 10 μl. MS/MS column was ZORBAX SB-C18 with mobile phase of acetonitrile-0.1% formic acid(80∶20,V/V)at flow rate of 0.2 ml/min;column temperature was 30 ℃;volume injection was 0.5 μl. Ionization mode was ESI+,atomization gas pressure was 25 psi,gas flow as 8.0 L/min,capillary voltage was 4 000 V,capillary outlet voltage was 120 V,precursor ion was 303 m/z,scan range was 50-500 m/z and the gas temperature was 350 ℃. RESULTS:The linear range of rosin acid was 2.5-100.0 μg/ml. RSDs of preci-sion,stability and reproducibility tests were lower than 2.0%,recoveries was 96.75%-98.11%(RSD=0.53%,n=6). CONCLU-SIONS:The method is simple,accurate and reproducible,and suitable for the content determination of rosin acid in Rheumatoid arthritis tablet.

OBJECTIVETo develop a screening program for spinal muscular atrophy (SMA) carriers, and to assess the carrier frequency and detection rate in Shanghai region.

METHODSQuantitative analysis of the SMN1 gene by real-time PCR was developed using specimens from 15 SMA patients and 76 SMA parents from 38 affected nuclear families. A pilot screening was carried out for 1741 asymptomatic pregnant women. Frequencies of SMN1 alleles were determined with the Hardy-Weinberg equilibrium.

RESULTSForty five out of the 1741 women were identified as SMA carriers by the presence of single copy of SMN1. The frequencies of no copy, 1 copy, 2 copy and 3 copy alleles were 1.37 U+00D7 10-2, 9.45 U+00D7 10-1, 2.80 U+00D7 10-2 and 1.27 U+00D7 10-2, respectively. The adjusted SMA carrier frequency was 1:35 with a detection rate of 94.49%. For those with a negative screening result, individuals with 3 copies carried a higher residual risk.

CONCLUSIONThe incidence of SMA carriers in Shanghai region is similar with that in Caucasian populations. Carrier screening has high detection efficiency. An effort should be made to further distinguish SMN1 gene copy numbers for those with more than 2 copies, since accurate determination of 2 and 3 copy allele frequencies is essential for post-screening genetic consulting.

Alleles ; Female ; Gene Dosage ; Gene Frequency ; Heterozygote ; Humans ; Male ; Muscular Atrophy, Spinal ; diagnosis ; genetics ; Pilot Projects ; Pregnancy ; Real-Time Polymerase Chain Reaction ; Survival of Motor Neuron 1 Protein ; genetics

Background The rapid diagnosis can win more treating opportunities for patients with fungal keratitis.Even though the fungal culture is the gold standard for the diagnosis of fungal keratitis,it is difficult in early diagnosis due to the long duration of cultivation and false-negative rate.Objective This trial was to explore the clinical value in the rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy.Methods Corneal scraping and laser scanning confocal microscopy were separately performed in 167 eyes of 167 patients with fungal keratitis.All the eyes were examined by the slit lamp,followed by laser scanning confocal microscope,and then the 10％ KOH corneal smear was examined under the optical microscope.Results The positive rate of diagnosis was 75％ (125/167) by corneal scraping,and that by laser scanning confocal microscopy was 91％ (152/167).The positive rate of examining outcome was significantly higher in laser scanning confocal microscopy than that of corneal scraping (x2 =14.88,P =0.00).The positive results were 114 cases and negative results were 4 cases by two methods,with the concordance rate 70.7％ (118/167).The hyphae or spore were seen in 32 cases by laser scanning confocal microscopy in 42 negative cases by corneal scraping,and in 15 negative cases by confocal laser scanning microscopy,11 positive outcomes were offered by corneal scraping.Conclusions The combined application of corneal scraping with confocal laser scanning microscopy can improve and speed up the diagnosis positive rate of fungal keratitis.

OBJECTIVETo evaluate the application of CXC chemokine receptor-4 (CXCR4) combined with alpha-methylacyl-CoA racemase (P504S) or P63 protein in the differential diagnosis of benign and malignant prostatic diseases.

METHODSThe EnVision immunohistochemical method was used to detect the expressions of CXCR4, P504S and P63 protein in 40 specimens of PCa not treated by any anticancer therapy and 30 specimens of BPH tissues. The correlation was analyzed between CXCR4 expression and the characteristics of PCa metastasis.

RESULTSOf the 40 cases of PCa, 33 (82.5%) were stained positive for CXCR4, 37 (92.5%) for P504S and 2 (5%) for P63 protein. Of the 30 cases of BPH, 5 (16.6%) exhibited positivity for CXCR4, 1 for P504S and all for P63. P504S + P63 showed a higher rate of correct diagnosis of PCa than either CXCR4 + P63 or P504S + CXCR4. There was a statistically significant correlation between CXCR4 expression and cancer metastasis (P < 0.05).

CONCLUSIONP504S, CXCR4 and P63 are useful tumor markers for the diagnosis and differentiation of benign and malignant prostatic diseases. CXCR4 gives a high rate of correct diagnosis when combined with P504S or P63, and has an important application value in the differential diagnosis of benign and malignant prostatic diseases.

Aged ; Aged, 80 and over ; Biomarkers, Tumor ; biosynthesis ; Diagnosis, Differential ; Humans ; Male ; Membrane Proteins ; biosynthesis ; Middle Aged ; Prostatic Hyperplasia ; diagnosis ; metabolism ; pathology ; Prostatic Neoplasms ; diagnosis ; metabolism ; pathology ; Racemases and Epimerases ; biosynthesis ; Receptors, CXCR4 ; biosynthesis

OBJECTIVETo observe the protective effect of Acanthopanax senticosus saponins (ASS) on myocardial ischemia-reperfusion injury in rats.

METHODThe myocardial ischemia-reperfusion model was induced by 30 min left anterior descending coronary occlusion and 120 min reperfusion in rats. The changes of myocardial infarct size (MIS), the serum creatine phosphokinase (CK) and lactate dehydrogenase (LDH) activity, the serum lipid peroxidation (LPO) content and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity and plasma endothelin (ET), angiotensin II (Ang II), prostacycline (PGI2) and thromboxane A2 (TXA2) levels and myocardial free fatty acid (FFA) content of infarct and noninfarct area were determined.

RESULTIn rats treated by ASS (in a dosage of 25, 50 and 100 mg x kg(-1) i.v. at 30 min after coronary occulusion), the MIS was significantly reduced, the serum CK and LDH activity, the plasma ET, Ang II and TXA2 level and myocardial FFA content declined, while plasma PGI2 level and PGI2/TXA2 was increased signficantly. In addition, serum LPO content declined, SOD and GSH-Px activity were increased markedly.

CONCLUSIONASS has protective effect on myocardial ischemia-reperfusion injury, which may be due to its function of improving free radicals and myocardial metabolism, decreasing plasma ET, Ang II and TXA2 levels and increasing plasma PGI2 level and PGI2/TXA2 ratio etc.

Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Eleutherococcus ; chemistry ; Female ; Male ; Myocardial Reperfusion Injury ; metabolism ; pathology ; Myocardium ; metabolism ; pathology ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology

BACKGROUND: The present study aimed to determine the short-term and long-term outcomes of critically ill patients with acute respiratory insufficiency who had received sedation or no sedation. METHODS: The data of 91 patients who had received mechanical ventilation in the first 24 hours between November 2008 and October 2009 were retrospectively analyzed. These patients were divided into two groups: a sedation group (n=28) and a non-sedation group (n=63). The patients were also grouped in two groups: deep sedation group and daily interruption and /or light sedation group. RESULTS: Overall, the 91 patients who had received ventilation ≥48 hours were analyzed. Multivariate analysis demonstrated two independent risk factors for in-hospital death: sequential organ failure assessment score (P=0.019, RR 1.355, 95％CI 1.051–1.747, B=0.304, SE=0.130, Wald=50483) and sedation (P=0.041, RR 5.015, 95％CI 1.072–23.459, B=1.612, SE=0.787, Wald=4.195). Compared with the patients who had received no sedation, those who had received sedation had a longer duration of ventilation, a longer stay in intensive care unit and hospital, and an increased in-hospital mortality rate. The Kaplan-Meier method showed that patients who had received sedation had a lower 60-month survival rate than those who had received no sedation (76.7％ vs. 88.9％, Log-rank test=3.630, P=0.057). Compared with the patients who had received deep sedation, those who had received daily interruption or light sedation showed a decreased in-hospital mortality rate (57.1％ vs. 9.5％, P=0.008). The 60-month survival of the patients who had received deep sedation was significantly lower than that of those who had daily interruption or light sedation (38.1％vs. 90.5％, Log-rank test=6.783, P=0.009). CONCLUSIONS: Sedation was associated with in-hospital death. The patients who had received sedation had a longer duration of ventilation, a longer stay in intensive care unit and in hospital, and an increased in-hospital mortality rate compared with the patients who did not receive sedation. Compared with daily interruption or light sedation, deep sedation increased the in-hospital mortality and decreased the 60-month survival for patients who had received sedation.

OBJECTIVETo analyse retrospectively the diagnosis and treatment of severe pneumonia in kidney transplantation recipients.

METHODSBetween January 1999 and December 2003, 172 adult patients underwent kidney transplantation at our department. In all severe pneumonia cases, empirical therapy was initiated with aztreonam, erythromycin and ganciclovir. And the therapy was switched to proper antibiotics according to the results of sensitivity testing. Responsible pathogen was detected by study of BAL (bronco-alveolar-lavage), sputum and blood specimen. Analyses included cell differential count, cytopathologic examination and cultures for bacteria, fungi and viruses. The immunosuppressive therapy was drastically reduced. Hypoxia was relieved by BiPAP (Bi-level Positive Airway Pressure) or mechanical ventilation if necessary.

RESULTSSeventeen cases (9.9%) of pneumonia were observed in the 172 recipients, only 11 (65%) patients experienced severe pneumonia, 1 (9%) of them died. Fever was the most common symptom on presentation (82%). On presentation 46% of the patients presented with classical clinical syndrome of fever accompanied by cough and dyspnea. Positive rate of BAL and blood culture were 100% and 46% respectively. BiPAP and mechanical ventilation were required in 6 and 2 cases respectively.

CONCLUSIONBAL is preferred for early detection of responsible pathogen. A combination of drastic reduction of the immunosuppressive regimen, implementation of appropriate empirical antibiotics, proper BiPAP or mechanical ventilation are important.

Adult ; Combined Modality Therapy ; Female ; Humans ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Pneumonia ; diagnosis ; etiology ; therapy ; Respiration, Artificial ; Retrospective Studies ; Transplantation, Homologous ; adverse effects

BACKGROUNDMuch research has been focused on ischemia/reperfusion injury (IRI) to the transplanted organs. As a free radical, nitric oxide (NO) plays an important role in IRI. In this study, the production of NO and its functions during IRI were monitored in rat models after allotransplantation of kidney grafts.

METHODSOf 75 male LEW rats, 30 served as donors, and the remaining 45 rats were divided into three groups (15 rats in each group): controls (group 1), kidney allotransplantation followed by bilateral nephrectomy during reperfusion (group 2), 2 hours before operation, donors and recipients were treated with N(G)-nitro L-arginine methyl ester (L-NAME), a NO synthase inhibitor, at a dose of 30 mg/kg (group 3). Bilateral nephrectomies were performed while kidney grafts were reperfused. The kidney grafts were hypothemically stored for 24 hours. The production of NO before and after reperfusion was measured by electron paramagnetic resonance (EPR). The creatinine level, the glomerular filtration rate (GFR) and the protein carbonyl content in tissue samples were recorded on the first and the fifth day after operation. The data were evaluated by one-way analysis of variance. Differences were considered to be statistically significant when a P value was less than 0.05.

RESULTSAfter reperfusion for 15 minutes, the production of NO increased remarkably and kept increasing till 120 minutes, after which the level returned to normal. In group 3, which was pretreated with L-NAME, creatinine levels were higher than those in group 2 at the 24th hour (4.10 +/- 0.50 mg/dl vs. 3.77 +/- 0.42 mg/dl, P < 0.05) and the 120th hour (3.19 +/- 0.79 mg/dl vs. 2.22 +/- 0.53 mg/dl, P < 0.05). GFR levels in group 3 were lower than those in group 2 at the 24th hour (0.50 +/- 0.12 ml/min vs. 0.71 +/- 0.19 ml/min, P < 0.05) and the 120th hour (0.59 +/- 0.38 ml/min vs. 1.27 +/- 0.23 ml/min, P < 0.01). The content of protein carbonyl in tissue samples of group 3 was lower than that in group 2 at the 24th hour (29.01 +/- 7.02 nmol/mg protein vs. 49.39 +/- 13.13 nmol/mg protein, P < 0.05), but was higher than that at the 120th hour (75.71 +/- 16.74 nmol/mg protein vs. 57.93 +/- 15.32 nmol/mg protein, P < 0.05).

CONCLUSIONSAfter transplantation of hypothemically stored kidney grafts, the increased NO production in the early stage has protective effects on the transplanted kidney. Application of L-NAME to inhibit NO production is harmful to the recovery of the renal functions of kidney grafts.

Animals ; Creatinine ; blood ; Electron Spin Resonance Spectroscopy ; Glomerular Filtration Rate ; Kidney Transplantation ; Male ; Nitric Oxide ; biosynthesis ; Oxidation-Reduction ; Proteins ; metabolism ; Rats ; Rats, Inbred Lew ; Reperfusion Injury ; metabolism

OBJECTIVETo study the effect of vitamin B on treatment of hyperhomocysteinemia and endothelial dysfunction in renal-transplant recipients.

METHODSThirty-six stable hyperhomocysteinemic renal-transplant recipients were randomly assigned to vitamin treatment (group A, n = 18, folic acid 5 mg/d, vitamin B(6) 50 mg/d, B(12) 1000 microg/d) or controlled group (group B, n = 18) for 6 months. All subjects underwent assessment of levels for creatinine, creatinine clearance, average pressure, total cholesterol, triglyceride and fasting homocysteine. Endothelial function was evaluated using high-resolution vascular ultrasound.

RESULTSThe levels of homocysteine markedly decreased in group A [(13 +/- 4) micromol/L vs (20 +/- 5) micromol/L, t = 5.3, P < 0.01] after treatment, whereas no significant changes were observed in group B. In group A, endothelium dependent [(12 +/- 5)% vs (9 +/- 5)%, t = 2.9, P < 0.01] and independent [(18 +/- 4)% vs (12 +/- 5)%, t = 3.4, P < 0.01] vasodilatation responses significantly increased after treatment, no significant changes were observed in group B. Endothelium dependent [(9 +/- 6)%, t = 2.8, P < 0.01] and independent [(12 +/- 5)%, t = 3.5, P < 0.01] vasodilatation responses of group A were significantly lower than that of group B after treatment.

CONCLUSIONSVitamin B supplementation can reduce the levels of homocysteine and improve the endothelial function in hyperhomocysteinemic renal-transplant recipients.

Adult ; Drug Therapy, Combination ; Endothelium, Vascular ; drug effects ; physiopathology ; Female ; Folic Acid ; administration & dosage ; Humans ; Hyperhomocysteinemia ; drug therapy ; physiopathology ; Kidney Transplantation ; Male ; Middle Aged ; Treatment Outcome ; Vitamin B 12 ; administration & dosage ; Vitamin B 6 ; administration & dosage

Amino Acid Sequence ; Base Sequence ; Genes, Bacterial ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Staphylococcus epidermidis ; enzymology ; genetics ; beta-Lactamases ; chemistry ; genetics