Recent researches indicate that ubiquitin-protea some pathway plays an important role in apoptosis regulation. Proteasome inhibitors induce apoptosis in many kinds of neoplastic cells, thus provide a great opportunity for exploring synergy of proteasome inhibitors and other apoptosis-inducing agents. In this study, the effect of the proteasome inhibitor Z-LLL-CHO combined with etoposide (VP16) on leukemic cell lines M-07e and TF-1 was investigated by MTT assay, trypan blue exclusion, flow cytometry and Western blot. The results showed that the combination of Z-LLL-CHO and VP16 was much more effective than either agents alone in promoting cytotoxicity in both cell lines evaluated. Accumulation of cells in S + G2/M phase of the cell cycle was observed in the cells treated with VP16 and Z-LLL-CHO alone, while apparent increase of sub-G0/G1 fraction was detected in cells treated with combination of the agents. The cleavage of Bcl-2 into a shortened 22 kD fragment was detected in M-07e cells exposed to either agents alone, and the fraction of 22 kD fragment was increased in the cells treated with combination of the agents. In conclusion, the combination of Z-LLL-CHO and VP16 enhanced their individual cytotoxic effect by inducing apoptosis, in which increase of S + G2/M fraction in cell cycle as well as the enhanced cleavage of Bcl-2 are the possible mechanism of the additive effect on leukemic cells by Z-LLL-CHO and VP16.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cysteine Endopeptidases ; Etoposide ; pharmacology ; Humans ; Leukemia ; pathology ; Multienzyme Complexes ; antagonists & inhibitors ; Oligopeptides ; pharmacology ; Proteasome Endopeptidase Complex ; Proto-Oncogene Proteins c-bcl-2 ; analysis

The ubiquitin-proteasome pathway is the principal mechanism for the degradation of short-lived proteins in eukaryotic cells. Recently, proteasome inhibitors have been shown to induce apoptosis in many kinds of human malignant cells. In this study, the mechanism of apoptosis induced by proteasome inhibitor in leukemic cells was examined. Evaluated by MTT assay, treatment of leukemic cells with Z-LLL-CHO, a reversible proteasome inhibitor, induced cell death in a dose-dependent manner. Appearance of the sub G(0)/G(1) fraction of cell cycle observed in flow cytometry assay suggested the induction of apoptosis, which was further proved by typical DNA ladder and morphological study. Western blot displayed the cleavage of bcl-2 into a shortened 22 kD fragment and the decrease in the levels of caspase-3 precursor. A highly sensitive colorimetric assay was employed and the elevation of caspase-3 activity was detected in both cell lines after treatment with Z-LLL-CHO. By comparison, these results showed that the leukemic cell line M-07e and KG-1a, which both express bcl-2 at a relative high level, had different susceptibility to undergo apoptosis induced by Z-LLL-CHO, which possibly due to their different levels of expression and activation of caspase-3 precursor, as well as their different degree of bcl-2 cleavage after treated by Z-LLL-CHO.

OBJECTIVETo sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR).

METHODSTo exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I. The virus were enriched through ultracentrifugation. Other nucleic acids, such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar.

RESULTSA 4834bp sequence of bocavirus I were assembled.

CONCLUSIONSISPA-PCR is an economical and efficient technique for sequence a virus complete genome.

Base Sequence ; Bocavirus ; genetics ; isolation & purification ; DNA Primers ; genetics ; Diarrhea ; virology ; Genome, Viral ; Humans ; Parvoviridae Infections ; virology ; Polymerase Chain Reaction

OBJECTIVETo identification and analysis Aichi virus from diarrhea and normal children in Lanzhou, and discuss the relationship between Aichi virus and Infant Diarrhea.

METHODSAccording to the literature published data, Using RT-PCR method to amplified Aichi virus 3CD fragment and the positive products were sequenced and determined, and made the alignment analysis between the nucleotide sequences of the amplified fragment with the known sequence of this virus.

RESULTSThere was 1 case detection of Aichi virus in the 46 hospitalized children with diarrhea and 299 children with diarrhea out-patients specifically, Overall detection rate was 0.06%, and there was no Aichi virus was detected in normal control children. 2 viral 3CD gene and the known reference strains of nucleotide sequences were 97%, while phylogenetic analysis showed that genotype of 2 viral belongs to the B.

CONCLUSIONSThere existed B Genotype of Aichi virus in China, and more research is needed to clarified the etiology and epidemiology of Aichi virus characteristics.

Child ; China ; Diarrhea ; virology ; Feces ; virology ; Humans ; Infant ; Kobuvirus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Picornaviridae Infections ; virology

OBJECTIVETo study the epidemiologic characteristics of virus-induced acute diarrhea in children under 5 years old in Taiyuan, Shanxi province.

METHODSStool specimens and clinical data were collected from 346 inpatients with acute diarrhea from children less than 5 years old. Rotavirus-positive specimens were identified by ELASA kit. Calicivirus and astrovirus were detected by reverse transcription-polymerase chain reaction (RT-PCR). Adenovirus was done by polymerase chain reaction (PCR).

RESULTSOf the 346 specimens, the percentage of samples with Rotavirus, Calicivirus, Astrovirus, and Adenovirus was 40.8%, 7.5%, 6.4% and 3.2%. Among 141 rotavirus positive samples, serotype G1 (42.6%) was the predominant strain. More than 95% of viral diarrhea patients under hospitalization occurred among children younger than 2 years.

CONCLUSIONRotavirus is the major pathogen contributing to the acute diarrhea. The disease generally peaks at autumn/winter. The predominant rotavirus strain circulated was G1P[8].

Age Distribution ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Female ; Humans ; Infant ; Male ; Virus Diseases ; epidemiology ; virology ; Viruses ; classification ; genetics ; isolation & purification

OBJECTIVETo analyze factors influencing the choice of atrial septal occluder (ASO) for transcatheter closure of patients with secundum atrial septal defect (ASD).

METHODSA total of 1114 ASD patients [388 males, aged from 2 to 75 years, mean age (26.3 +/- 17.0) years] were enrolled. Patients were divided to adult (> 14 years, mean 34.4 years, n = 779) and child (< or = 14 years, mean 7.3 years, n = 335) groups. ASD size in different ultrasound cross-sections was determined by transthoracic echocardiography (TTE). ASO size was chosen on the basis of the maximum diameter of the defect (MD). Defect-shapes and rim lengths of ASD, the difference choice of ASO in the two groups were compared.

RESULTSMD of the defects ranged from 5 to 40 mm [mean (19.7 +/- 7.8) mm]. ASD was successfully occluded in 1085 out of 1114 patients (97.4%). Occluder size ranged from 6 to 46 mm [mean (25.8 +/- 8.9) mm] and the difference between occluder size and MD ranged from 2 to 10 mm [mean (6.1 +/- 3.4) mm, ASO/MD ratio 1.3:1]. Though the diameter of the defect was similar between the 2 groups, the size of occluder was significantly larger in adult group than that in child group (ASO/MD ratio 1.1 - 1.6:1 vs. 1.2 - 1.8:1, P < 0.05). MD was significantly correlated with ASO in both groups (r = 0.911 and r = 0.944 in adults and child groups, respectively, all P < 0.01). The size and increment of the occluder used in patients with deficient anterior rims was significantly bigger than patients with sufficient anterior rims (P < 0.01).

CONCLUSIONThe maximum diameter of the defect was the major determinant for selecting occluder size and choice of occluder size was also influenced by patient age, defect-shape and defect rim for transcatheter closure of secundum ASD.

Adolescent ; Adult ; Age Factors ; Aged ; Cardiac Catheterization ; instrumentation ; Child ; Child, Preschool ; Echocardiography ; Female ; Heart Septal Defects, Atrial ; therapy ; Humans ; Male ; Middle Aged ; Physicians ; psychology ; Practice Patterns, Physicians' ; Prosthesis Design ; Septal Occluder Device ; Young Adult

Group A rotavirus are the most frequently detected viral agent associated with the acute diarrhea in calves. In order to investigate the situation of rotavirus strains circulating in diary farms, a total of 117 fecal specimens were collected from diarrhea calves under 4 weeks-age on Yinluo diary farm in Daqing region in China from 2008 to 2009. Ten specimens were detected to be positive by a Rotavirus Group A Diagnostic Kit, which confirmed that the rotavirus was important viral agent associated with diarrhea in this diary farm. Based on the new classification system, G10P[11] genotype was determined in rotavirus positive samples. Sequence and Phylogenetic analysis indicated DQ-75 strain was introduced into our country with imported bovine.
Animals ; Cattle ; China ; Genotype ; Phylogeny ; Rotavirus ; classification ; genetics ; Rotavirus Infections ; virology

From November 2008 to January 2009, a sharp increase of diarrhea in children in Guangdong province appeared, we randomly collected 53 stool specimens from out-patient children with dirrhea in 3 major hospitals (Guangzhou City Children's Hospital, Shenzhen Baoan District Maternal and Child Health Hospital, First Affiliated Hospital of Shantou University). Rotavirus and calicivirus were screened by ELISA and RT-PCR. We found 29 cases of rotavirus infection with diverse serotypes. Only four cases were identified as calicivirus infection. The result indicated that rotavirus was a major pathogen of this high incidence of diarrhea from November 2008 to January 2009 in Guangdong Province.
Age Distribution ; Child, Preschool ; China ; epidemiology ; Diarrhea ; diagnosis ; epidemiology ; virology ; Diarrhea, Infantile ; diagnosis ; epidemiology ; virology ; Feces ; virology ; Humans ; Incidence ; Infant ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; diagnosis ; epidemiology ; virology ; Serotyping

Coptidis Rhizoma-Evodia Fructus is a classic herb pair in traditional Chinese medicine prescriptions, the famous prescription is called Zuojinwan, which comes from Danxi Xinfa, is composed of Coptidis Rhizoma-Evodia Fructus (6∶1). In this formula, Coptidis Rhizoma has the effect of clearing heat and drying diuresis, purging fire to remove toxin and clearing heart. Evodia Fructus has the effects of expelling cold and alleviating pain, checking upward adverse flow of Qi tostop vomiting, and assisting yang to stop diarrhea. Coptidis Rhizoma has the properties of bitter and cold, and Evodia Fructus has the properties of pungent and calidus. Pungent drugs have divergent effects, and bitter drugs have sedimentation effect, when used in combination, they can clear the liver and purge fire, calm the adverse-rising energy and stop vomiting. On the basis of Zuojinwan, Coptidis Rhizoma-Evodia Fructus medicine has derived different compatibility ratios. Different ratios are different in terms of efficacy, usage, clinical application. Although with the application of modern analytical instruments and the development of molecular pharmacology theory, the chemical constituents and Pharmacological effects of Coptidis Rhizoma and Evodia Fructus have been fully studied, as to the principle of compatibility, and the study of pharmacological effects and chemical constituents after the compatibility of the two drugs in different proportions, there is still no comprehensive system summary. This article makes a systematic and comprehensive explanation of Coptidis Rhizoma-Evodia Fructus from the aspects of famous literature, chemical composition, pharmacological effects and clinical applicationthrough querying literature and ancient books. In order to make this herb pair more standardized, and provide reference materials for further research and development for this herb pair.

This study was to investigate the protective effect of paeoniflorin (PF) on hydrogen peroxide-induced injury. Firstly, "SMILES" of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets. Injury-related molecules were obtained from GeenCards database, and the predicted targets of PF for injury treatment were selected by Wayne diagram. For mechanism analysis, the protein-protein interactions were constructed by String, and the KEGG analysis was conducted in Webgestalt. Then, cell viability and cytotoxicity assay were established by CCK8 assay. Also, the experimental cells were allocated to control, model (200 μmol·L