Adult ; Diverticulum ; complications ; Echocardiography ; methods ; Heart Diseases ; diagnosis ; Heart Ventricles ; pathology ; Humans ; Male ; Young Adult

Astragali Radix was firstly recorded in the "Shen Nong's Herbal Classic" as a top-grade and commonly used traditional Chinese medicine. Its frequently used slices include raw Astragali Radix and honey-processed products. In current studies, many reports were made on honey-processed Astragali Radix, whereas fewer study reports were made on the cutting process of Astragali Radix. Currently, because Astragali Radix is primarily cut by drug workers according to their operating experience, but with out specific cutting parameters, it is easy to cause the loss or mildew of active ingredients. As a result, the quality of Astragali Radix circulated in the market is not guaranteed, and the quality of their slices and preparations are hard to be controlled, which seriously impact the clinical efficacy. In response, this experiment was performed, in which the optimum cutting process of Astragali Radix was taken as the study objective, the Box-Benhnken central composite design in the response surface analysis was adopted, and the content and appearance character of astragaloside and calycosin-7-glucoside were regarded as the study indicators. Three factors, namely the softening time, the drying temperature and the drying time, were selected to optimize the cutting process of Astragali Radix and obtain the optimum cutting process parameters as follows: the softening time was 3 hours, the drying temperature was 50 degrees C, and the drying time was 4 hours. According to the verification test, the Astragali Radix cutting process is steady and feasible, which has certain significance for normalizing the cutting process of Astragali Radix.
Astragalus Plant ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Plant Roots ; chemistry

The study was aimed to investigate the application value of interphase fluorescence in situ hybridization (FISH) on cell smears in hematological diseases. Both interphase FISH on peripheral blood smears and bone marrow smears treated by methanol/acetic acid, and routine interphase FISH of bone marrow cells dropped on slides were done at the same time, in order to detect Ph chromosome by BCR/ABL dual color, dual fusion probe in 20 patients with chronic myelogenous leukemia or acute lymphoblastic leukemia which had been proven to display Ph chromosome positive. The results indicated that as compared with routine interphase FISH, the interphase FISH on cell smears could also offer reliable result. It is concluded that interphase FISH on cell smears is a kind of reliable and time-saving technique, which is also suitable for retrospective research and worthy to further apply in clinic.
Adult ; Aged ; Cytogenetic Analysis ; methods ; Female ; Hematologic Diseases ; diagnosis ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Interphase ; genetics ; Male ; Middle Aged ; Young Adult

10.3969/j.issn.1000-3606.2013.06.005

OBJECTIVETo determine the effect of butylphthalide on the expressions of protein disulfide isomerase (PDI) and P53 in the brain tissue of rats with Alzheimer's disease (AD).

METHODSSixty male adult rats were randomly divided into AD model group, butylphthalide group and control group (n = 20). AD models were established by injecting beta-amyloid protein 1-42 into the hippocampus of rats. Sixty days later, the learning and memory abilities of the rats were evaluated using Y-maze test, and the expressions of PDI and P53 in the brain tissue of the rats were measured by immunohistochemistry.

RESULTSCompared with the control group, the rats in AD model group exhibited significantly reduced learning and memory abilities, lowered expressions of PDI in the hippocampus and increased expression of P53 in the cortex (P > 0.01). In comparison with the model group, the rats in the butylphthalide group showed significantly increased PDI-positive cells in the hippocampus and decreased expression of P53 in the cortex (P < 0.01).

CONCLUSIONButylphthalide improves the learning and memory abilities of rats with experimental AD, the mechanism of which may involve inhibition of P53 expression and enhancement of PDI expression in the brain tissues.

Alzheimer Disease ; physiopathology ; Animals ; Apoptosis ; drug effects ; Benzofurans ; pharmacology ; Brain ; enzymology ; metabolism ; Disease Models, Animal ; Learning ; drug effects ; Male ; Memory ; drug effects ; Protein Disulfide-Isomerases ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Suppressor Protein p53 ; metabolism

The objective of this study was to investigate the prognosticating value of multiparameter flow cytometry in detection of minimal residual disease (MRD) and relapse risk of patients with acute myeloid leukemia (AML). Multiparameter flow cytometry (MPFC) analysis was used to detect the leukemia-associated aberrant immunophenotype (LAIP) of the pretreated patients with AML and to assess the levels of MRD after remission induction (Post-Ind MRD) and consolidation therapy (Post-Cons MRD). The results showed that the definite LAIP could be detected in 94.3% of the patients (115/122) with AML (except APL). Among 115 cases only one LAIP was identified in 15 cases (13.0%), but two or more LAIP were identified in other 100 cases (87.0%). The most frequent LAIP identified was cross-lineage antigen expression (40.9%). The percentages of asynchronous antigen expression, antigen over-expression and antigen lack expression were 20.9%, 27.0%and 34.8% respectively. MRD frequency was monitored in 41 AML patients with CR after remission induction chemotherapy and 2 or more cycles of consolidation chemotherapy. 24 patients were Post-Ind MRD(+) and 17 patients were Post-Ind MRD(-). The percentages of relapse in cases of Post-Ind MRD(+) and Post-Ind MRD(-) were 75.0% (18/24) and 29.4% (5/17) respectively after consolidation chemotherapy. The relapse free survival (RFS) times of the patients with Post-Ind MRD(+) and Post-Ind MRD(-) were 49.06 +/- 6.53 months and 11.92 +/- 1.64 months (p < 0.0001) respectively. 18 patients were Post-Cons MRD(+) and 23 patients were Post-Cons MRD(-). The percentages of relapse in cases of Post-Cons MRD(+) and Post-Cons MRD(-) patients were 100% (18/18) and 21.7% (5/23) respectively after consolidation chemotherapy. The RFS times of the patients with Post-Cons MRD(+) and Post-Cons MRD(-) were 41.74 +/- 5.52 months and 10.06 +/- 1.72 months (p < 0.0001) respectively. It is concluded that the levels of post-Ind MRD and post-Cons MRD identified in the patients with AML was highly associated with their RFS. The detection of MRD by MPFC provides prognostic information in AML patients.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Flow Cytometry ; methods ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; pathology ; Prognosis ; Recurrence ; Young Adult

OBJECTIVETo study effects of Yishen Kangxian Compound (YKC) and benazepril containing serums on HK-2 cells (human renal proximal tubule epithelial cells) in the process of renal tubular epithelial cells to mesenchymal myofibroblasts transdifferentiation (TEMT) by gene chip.

METHODSYKC and benazepril containing serums were prepared. Their inhibitory effects on HK-2 cells in the transforming growth factor-beta1 (TGF-beta1)-induced TEMT process were observed. HK-2 cells were randomly divided into four groups, i.e., the blank control group, the model group, the benazepril group, and the YKC group. The gross RNAs were extracted and purified by taking advantage of the HumanHT-12 v4 of IlluminaBeadChip. Differentially expressed genes were obtained after they were reversely transcribed to cDNA, incorporating biotin labeling probe, hybridized with GeneChip, picture signals of fluorescence in gene array scanned and compared with differential genes by computer analysis.

RESULTSDifferentially expressed genes were successfully identified by gene chip. Compared with the model group, there were 227 differentially expressed genes in the benazepril group, including 118 up-regulated genes and 109 downregulated genes. Compared with the model group, there were 97 differentially expressed genes in the YKC group, including 69 up-regulated genes and 28 down-regulated genes. The Gene Ontology (GO) analysis indicated that YKC was more actively involved in the regulatory process than benazepril in terms of cell damage, apoptosis, growth, NF-KB, protein kinase, neuron, and blood vessel growth.

CONCLUSIONSYKC and benazepril could inhibit the TEMT process of HK-2 cells. But YKC also had taken part in cell damage, apoptosis, growth,and more pathways of early stage TEMT.

Cell Line ; Cell Transdifferentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; Genomics ; Humans ; Kidney Tubules, Proximal ; cytology ; pathology

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Medicine, Tibetan Traditional ; Phyllanthus emblica ; chemistry ; classification ; Quality Control ; Tannins ; analysis ; Tibet

Objective To construct eukaryotic enhanced green fluorescent protein (EGFP) expressing recombinant plasmid, pEGFP-C1-hnRNP A1, which contains coding sequence of human hnRNP A1 (heterogeneous nuclear ribonucleo-protein A1), and to perform cellular localization analysis of EGFP tagged hnRNP A1 under stress. Methods Total RNA was isolated from HeLa cell used for synthesis of first-strand cDNAs using reverse primers that are specific for the 3′-un-translated region of hnRNP A1. hnRNP A1 gene fragments were then amplified by touch-down PCR from those cDNAs and inserted into pEGFP-C1 fluorescent bearing vector through EcoRⅠ/BamHⅡdouble enzyme digestion and T4 DNA Ligase connection. The recombinant pEGFP-C1-hnRNP A1 plasmid was transfected into HeLa cells and green fluorescent tagged fusion proteins was examined by Western blot and confocal fluorescence microscopy. Co-localization of EGFP-hnRNP A 1 with poly (A)+mRNA (the marker of the stress granules), or DCP1a (the marker of processome) were detected by RNA fluores-cence in situ hybridization and immunofluorescence. Results The pEGFP-C1-hnRNP A1 was sequenced and digested cor-rectly by restriction single/double enzyme. The green fluorescent fusion protein was also detected in transfected HeLa cell by Western blot and confocal fluorescence microscopy. EGFP-hnRNP A1 co-localizes with poly(A)+mRNA, but not DCP1a. Conclusion Recombinant eukaryotic plasmid of pEGFP-C1-hnRNP A1 was constructed successfully and expressed effec-tively. EGFP tagged hnRNP A1 takes part in forming stress granules.

Objective To purify human 14-3-3β (YWHAB) recombinant protein expressed in the E.coli, prepare its antiserum and construct the eukaryotic expression vector for transfecting mammalian cells. Methods The human 14-3-313 recombinant protein expression vector pET30a (+) /YWHAB constructed by the ORF of YWHAB gene and prokaryotic expression vector pET30a (+) was transformed into E.coli BL21 (DE3). The expression of the recombinant protein was induced by IPTG and the protein was purified by affinity chromatography on a Ni-NTA resin. BALB/c mice were immunized by the purified protein, and ELISA and Western blotting were employed to detect the titer and specificity of the antiserum. The open reading flame of YWHAB gene was obtained by PCR, the purified PCR product digested by BamH Ⅰ and EcoR Ⅰ was cloned into the eukaryotic expression vector pEGFP-N1, and the product digested by BamH Ⅰ and Hind Ⅲ was cloned into the eukaryotic expression vector pCDNA3.1 (+). The recombinant vectors were identified by PCR and enzyme digestion. Results The recombinant protein was expressed as a soluble protein with a relative molecular mass of about 32 kD, which was consistent with the expected value. The recombinant protein was purified using affinity chromatography to yield a purity up to 90%. The antiserum had high specificity and titer (1: 50000). The results of PCR and enzyme digestion verified successful construction of the eukaryotic recombinant expression vector pEGFP-N1/YWHAB and pCDNA3.1 (+)/YWHAB. Conclusion The recombinant human 14-3-3β protein, the antiserum and the eukaryotic expression vector obtained may facilitate further functional study in the human 14-3-3β protein.