A simple and effective method for the duplex PCR detection was developed by using sequences of exogenous gene(RDV MP-)and endogenous gene(Zein)as templates for PCR amplification.The results of routine PCR amplification for RDV MP-gene in transgenic maize suggested that RDV MP-gene can stably inheritate in transgenic plants and their progenies;The duplex PCR detection of all negative and part positive samples that obtained by routine PCR amplification confirmed that above negative results were exact,also showed that the quality of extracted DNA can meet the need of PCR amplification.The error ratio of negative samples was 1.4%.The method used in this study was simple and credible and can be used to detect transgenic plants and their products.

Objective To establish a novel method to determine the absolute content of quercetin by proton nuclear magnetic resonance (qNMR).Methods DMSO-d6 was employed as solvent,and maleic acid as an internal standard.Proton signal peaks at δ7.50-7.58 and δ6.26 of maleic acid were served as quantification peaks.The content of quercetin is determined with qNMR in comparison with the results obtained by mass balance method.Results Linear regression of quantitative peak areas ratio (As/Ar) of quercetin-maleic acid vs mass ratio (ms/mr) yielded a correlation coefficient of 0.999 3 and a regression equation ofy =2.963 x + 0.134 1.The contents of three batches quercetin were 85.20％,84.93％,and 85.27％,the average was 85.13％ and its RSD was 0.21％.The results were generally consistent with that of mass balance methods.Conclusion This method was easy and simple to handle,and the analysis results were accurate.It could be the complementary for the mass balance method.

Objective: To investigate the killing activity of cytokine-induced killer (CIK) cells after incubation with autologous tumor cell lysate-pulsed dendritic cells (Ag-DC) and to evaluate the immune functions of patients, the clinical efficacy and side effect of Ag-DC in combination with CIK on hepatocellular carcinoma. Methods: Peripheral blood mononuclear cells were isolated from 24 patients with hepatocellular carcinoma, and cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4 to produce dendritic cells (DCs). The DCs were pulsed with autologous tumor cell lysate. T lymphocytes from PBMC were cultured with interferon-Y (IFN-γ), IL-2, CD3-moAb, and IL-1α to prepare CIK. The killing effect of CIK on SMMC-7721 was investigated after CIK was incubated with Ag-DC. After immunotherapy with Ag-DC and CIK, immunolog-ic and clinical responses of the 24 patients were evaluated. Results: The killing effect of CIK was remarkably improved after CIK was incubated with Ag-DC. The immunotherapy with DC and CIK alleviated symptoms and improved the immune functions of the patients. Except for transient fever and chill, no remarkable adverse events were observed. Conclusion: Ag-DC in combination with CIK shows short-term efficacy on hepatocellular carcinoma through inducing specific anti-tumor immunity and can be an effective adjuvant therapy for hepatocellular carcinoma.

The deep fermentation technique of Tricholoma matsutake is systemically studied in this paper firstly. The best culture determined by orthogonal test is 3g/L of cornflour, 1g/L of glucose, 1g/L of bean cake flour, 1mL/L of corn steep liquid, 1g/L of KH 2PO 4. The best fermenting condition is: 25℃, rotating speed 160 r/min, pH5.0,inoculating amount 10%, 120mL culture medium per 500mL flask. Under these conditions, the mycelia reach 12.94g/L after fermenting 12d.

Objective To improve the understanding of cutaneous intravascular natural killer/T-cell lymphoma (CIVNKTC). Methods Clinical data on five cases of CIVNKTC were collected. The histopathological feature, treatment and prognosis of CIVNKTC were retrospectively analyzed and discussed. Results Of the 5 patients, 1 was male and 4 were female. The age of onset ranged from 38 to 83 years (average, 56.2 years). All the patients presented with multiple plaques and nodules as the primary symptoms. Histopathological examination revealed vasodilatation in the dermis and subcutaneous tissue, as well as atypical lymphoid cells with large hyperchromatic nuclei containing 1-2 small nucleoli in dilated veins. Immunohistochemical studies of tumor cells showed positive staining for CD3ε, cytotoxic proteins (including T cell-restricted intracellular antigen-1, granzyme B and perforin)and Epstein-Barr virus(EBV)-encoded microRNA, but negative staining for cytokeratin, CD20, CD79a, CD4 and CD8. Furthermore, the tumor cells stained positive for CD56 in two patients. Among the 5 patients, only 2 received chemotherapy and the remaining received no treatment. During a 24-month follow-up, 4 patients died, and only 1 survived with the tumor. Conclusion CIVNKTC is a rare extranodal Hodgkin′s lymphoma with distinct histologic manifestations and immunophenotypes, rapid and aggressive clinical course, and poor prognosis.

Adult ; Aged ; Combined Modality Therapy ; Electric Stimulation Therapy ; Electroacupuncture ; Essential Tremor ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective To investigate the inhibitory effect of carbon-coated iron nanoparticles carrying cisplatin on the growth of NCI-H446 lung cancer cells and expressions of Caspase 3 and Survivin mRNA.Methods NCI-H446 lung cancer cells were treated with iron-carbon nanoparticles and/or cisplatin.The cell viability was detected by MTT method,and the mRNA expressions of Caspase 3 and Survivin were measured with RT-PCR.Results Cisplatin could inhibit the growth of NCI-H446 lung cancer cells,and the inhibitory effect was stronger when it was combined with the iron-carbon nanoparticles.The cells had apoptosis.The mRNA expression of Caspase 3 of NCI-H446 lung cancer cells was remarkably enhanced after treatment with iron-carbon nanoparticles combined with cisplatin,while the mRNA expression of Survivin was notably weakened (P＜0.05).Conclusion Carbon-coated iron nanoparticles carrying cisplatin could significantly increase the chemotherapy sensitivity of cisplatin on NCI-H446 lung cancer cells and enhance the therapeutic efficacy of chemotherapy drugs.

Objective To compare the diagnostic yield of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for solid pancreatic masses performed with three different needle types through the cytological results.Methods All patients with solid pancreatic masses larger than 2cm from December 2010 to May 2011 were enrolled,and divided into two groups according to different access of EUS-FNA,trans-gastric approach with 19-,22-and 25-gauge needles (n =42) and trans-duodenal approach with 22-and 25-gauge needles (n =10).In both groups,EUS-FNA was performed with randomization of needle types.During the puncture,the suction,the number of movements,and the depth of insertion were fixed.At the end of the puncture,a liquid-based cytological (LBC) preparation was used to fix the specimen.One cytopathologists was assigned to make the diagnosis and comparison.Results Technical success was 100％ and no procedure related complications occurred.No statistically significant differences were observed in different needles in terms of all cytological parameters between two groups (P ＞ 0.05).However,the 25-gauge needle showed a trend towards a higher sensitivity,specificity,positive predictive value,negative predictive value and accuracy.Conclusion There is no significant difference in yield of cytological results between different needle types,although 25-gauge needle shows a relative superiority.

Fifteen cassaen-type diterpenes were isolated from the 95% ethanolic extract of the seeds of C. minax through various chromatographic techniques. Their structures were identified on the basis of spectroscopic data as pulcherralpin (1), caesalpinin ML (2), chamaetexane C (3), chamaetexane D (4), 6β, 18-diacetoxycassan-13, 15-diene (5), neocaesalpin K (6), neocaesalpin MP (7), neocaesalpin M (8), neocaesalpin Q (9), neocaesalpin P (10), neocaesalpin R (11), caesaldekarin D (12), caesaldekarin A (13), caesaldekarin b (14), 3β,6α-diacetoxyvouacapane (15). Among them, compounds 14, 9-11 were isolated from this plant for the first time.
Caesalpinia ; chemistry ; Diterpenes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Seeds ; chemistry ; Spectrometry, Mass, Electrospray Ionization

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.