Animals ; Antioxidants ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Fatigue ; prevention & control ; Male ; Mitochondria, Heart ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Panax ; chemistry ; Physical Exertion ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley

Animals ; Antioxidants ; pharmacology ; Female ; Hypoxia ; physiopathology ; Male ; Mice ; Panax ; chemistry ; Random Allocation ; Reperfusion Injury ; prevention & control ; Saponins ; isolation & purification ; pharmacology

Animals ; Antioxidants ; pharmacology ; Brain ; metabolism ; Brain Ischemia ; metabolism ; Male ; Malondialdehyde ; metabolism ; Panax ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism

Our previous study demonstrated that human KIAA0100 gene is a novel acute monocytic leukemia-associated antigen (MLAA) gene. But the functional characterization of human KIAA0100 gene has remained unknown to date. Here, firstly, bioinformatic prediction of human KIAA0100 gene was carried out using online software;Secondly, human KIAA0100 gene expression was downregulated by the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system in U937 cells. Cell proliferation and apoptosis were next evaluated in KIAA0100-knockdown U937 cells. The bioinformatic prediction showed that human KIAA0100 gene was located on 17q11.2, and human KIAA0100 protein was located in the secretory pathway. Besides, human KIAA0100 protein contained a signal peptide, a transmembrane region, three types of secondary structures (alpha helix, extended strand, and random coil) , and four domains from mitochondrial protein 27 (FMP27). The observation on functional characterization of human KIAA0100 gene revealed that its downregulation inhibited cell proliferation, and promoted cell apoptosis in U937 cells. To summarize, these results suggest human KIAA0100 gene possibly comes within mitochondrial genome; moreover, it is a novel anti-apoptotic factor related to carcinogenesis or progression in acute monocytic leukemia, and may be a potential target for immunotherapy against acute monocytic leukemia.

Monoclonal antibodies (MAbs) are important tools for the study of proteins′ function and structure. But there has been no report on the preparation of MAbs against human KIAA0100 protein up to date. Here, first, we generated the mouse MAb against human KIAA0100 protein using purified recombinant 6×Histidinc (6×His)- tagged human KIAA0100 protein segment (1557–2234) as an antigen; then, the mRNA expression of human KIAA0100 gene was detected in U937 cells using Northern blot analysis. The results showed that the mouse MAb against human KIAA0100 protein could sensitively recognize the human KIAA0100 protein using Western blot analysis and immunocytochemistry analysis. Besides, Western blot analysis revealed that human KIAA0100 gene possibly encoded two different protein products (254 kDa and < 250 kDa) in U937 cells. Moreover, Northern blot analysis confirmed that human KIAA0100 gene might produced two different mRNA products (6000–10000 bp and 5000–6000 bp) in U937 cells. The results provide a basis for large-scale production of the MAb against human KIAA0100 protein, which will be useful for the study of human KIAA0100 protein′s function/structure and MAb-targeted drugs in the future.

Bacterial Infections ; epidemiology ; microbiology ; China ; Drug Resistance, Bacterial ; Humans ; Microbial Sensitivity Tests

Animals ; Brain ; metabolism ; Brain Ischemia ; metabolism ; Excitatory Amino Acids ; metabolism ; Female ; Male ; Panax ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Saponins ; isolation & purification ; pharmacology

OBJECTIVE:To establish a method for the determination of dissolution of Xiaocaihu pill,and compare the differ-ence of preparation from different manufacturers. METHODS:Using 0.1 mol/L HCl as dissolution medium,rotating basket method was used to determine the dissolution of preparations. HPLC was adopted to determine the content of baicalin:column was TSKgel ODS C18 with mobile phase of methanol-water-phosphoric acid (65∶35∶0.7,V/V/V) at a flow rate of 1 ml/min,detection wave-length was 280 nm,column temperature was 30 ℃,and injection volume was 5 μl. RESULTS:The linear range of baicalin was 0.488-124.8 mg/L (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recovery was 100.14%-104.78%(RSD=1.58%,n=9). The average t50(50% dissolution time)of baicalin was 85.81 min. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and can be used for the dissolution determination of Xiaocaihu pill. Xiaocaihu pill from different manufacturers shows great differences,both preparation formulation and clinical use should attach importance to the dissolution of solid preparations.

Objective To evaluate the application value of 18 F‐FDG PET/CT in patients with fever of unknown origin (FUO) to improve the etiological diagnosis accuracy of PET/CT in patients with FUO and provide better service for clinical prac‐tice .Methods Clinical data and imaging results of 144 cases of FUO patients diagnosed through 18 F‐FDG PET/CT examination in Wuhan union hospital from December 2009 to December 2011 were retrospectively analyzed .The final diagnoses were based on sur‐gical pathology ,aspiration cytology ,or discharge diagnosis combined with more than 12 months of follow‐up results .All the PET/CT images were analyzed and the clinical application value of PET/CT in FUO patients were evaluated through the diagnose rate , diagnose efficiency and clinical staging of malignant tumor .Results Among the 144 patients ,there were 130 cases ultimately had clear diagnosis ,which including 41 cases of malignant tumor ,89 cases of benign lesions and 14 cases the cause of unknown .PET/CT detected abnormal lesions for 97 .6% (40/41) of patients with malignant lesions and 88 .7% (79/89) of patients with benign le‐sions ,and the diagnostic sensitivity ,specificity ,accuracy ,positive predictive value ,and negative predictive value were :97 .6% (40/41) ,86 .5% (77/89) ,90% (117/130) ,76 .9% (40/52) and 98 .7% (77/78) .PET/CT detected the primary lesions for the first time in 46 .3% (19/41) patients with malignant tumor ,and detected more lesions than the traditional imaging methods in 92 .7% (38/41) of these patients .Conclusion 18 F‐FDG PET/CT imaging can provided important reference value for the detection of primary lesions ,the qualitative diagnosis of benign and malignant lesions and clinical staging of malignant lesions .