Acupuncture Therapy ; Adult ; Combined Modality Therapy ; Computers ; utilization ; Female ; Hand Injuries ; therapy ; Humans ; Male ; Middle Aged ; Ultrasonic Therapy

China ; epidemiology ; Humans ; Patient Acceptance of Health Care ; Tuberculosis, Pulmonary ; epidemiology ; psychology

OBJECTIVETo investigate the in vivo functional roles of the La autoantigen (La), the human homologue of the 33-kDa vesicle-associated membrane protein-associated protein (hVAP-33), and the subunit gamma of the human eukaryotic initiation factors 2B (eIF2Bgamma) as co-infection factors supporting chronic infection with hepatitis C virus (HCV).

METHODSSmall interfering (si)RNAs were designed against the HCV internal ribosome entry site (IRES) and transfected into Huh7 cells chronically infected with the HCV pseudovirus (designated as Huh7-HCV cells). The IRES siRNA producing the most effective silencing was selected for further analysis by fluorescence quantitative polymerase chain reaction (qPCR). siRNAs designed against La, hVAP-33, and eIF2Bgamma and the IRES-specific siRNA were then transfected, respectively or in various combinations, into the Huh7-HCV cell line for 48 h. The delta CT values were calculated and used to compare the HCV inhibitive efficacies of the siRNAs in isolation or in combination. Western blotting analysis was used to compare the quantity of core protein expression in each group.

RESULTSThe four gene-specific siRNAs, in isolation or in combination, caused inhibition of HCV replication and gene and protein expressions to varying degrees. The combination of La + IRES siRNAs produced the strongest inhibition of HCV core antigen expression. The combinations of hVAP-33 + IRES siRNAs and eIF2Bgamma + IRES siRNAs produced stronger inhibitions of HCV replication and gene and protein expressions than either hVAP-33 siRNA or eIF2Bgamma siRNA alone.

CONCLUSIONLa, hVAP-33, and eIF2Bgamma act as co-infection factors of HCV chronic infection in vivo. HCV replication and gene and protein expression can be inhibited significantly by RNA interference of these co-infection factors and/or HCV IRES.

Autoantigens ; genetics ; Cell Line ; Eukaryotic Initiation Factor-2B ; genetics ; Hepacivirus ; immunology ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Ribonucleoproteins ; genetics ; Vesicular Transport Proteins ; genetics ; Virus Replication

Objective　To explore the role of acute infection of Chlamydia pneumoniae (Cpn) in respiratory diseases. Methods　Microimmunofluorescence test was used to detect IgG antibodies for Cpn in serum obtained from 93 inpatients and PCR was used to test Cpn in detection of Cpn DNA in throat specimens from 55 of the 99 patients. Results　Acute Cpn infection was diagnosed in 35.5% of the respiratory diseases. Antibodies for Cpn (titer of ≥512) were present in 47.6% of the pneumonia group, which may suggest that during 1998 to 1999, Cpn caused an epidemic in Beijing. They were also present in 50% of asthma group, 50.0% of pulmonary heart disease group and 26.3% of lung cancer group. Only five patients (9.1%) were positive by PCR. There exists discrepancy between serological and PCR results. Conclusion　Detection of IgG antibodies for Cpn conduces to diagnosis of acute Cpn infection and give advice for appropriate therapy.

OBJECTIVETo determine the serotype and genotype of a sample with ABO blood group discrepancies.

METHODSSerotype was determined with serological method. Sequence specific primer polymerase chain reaction (SSP-PCR) was carried out based on the serotype. Sequences of exons 6 and 7 of ABO gene was analyzed by sequence-based testing (SBT).

RESULTSCompletely agglutinated A antigen, half agglutinated B antigen and weak agglutinated anti-B antibody were detected in both erythrocytes and serum, which suggested presence of a ABw serotype. An A/Bw12 genotype was revealed by B subgroup detection. Sequences of exons 6 and 7 were 278CT, 297GA and 467CT, 526CG, 657CT, 703GA, 796CA, 803GC, 930GA, respectively. The genotype fit with A102/B101 except for a nt278 C>T mutation. Blood group antigen gene mutation database (BGMUT) search has confirmed the mutant allele to be Bw12.

CONCLUSIONAn A102/Bw12 genotype has been found in the Chinese population.

ABO Blood-Group System ; genetics ; Base Sequence ; Blood Group Antigens ; genetics ; Blood Grouping and Crossmatching ; methods ; Female ; Genotype ; Humans ; Middle Aged ; Molecular Sequence Data ; Mutation

OBJECTIVETo investigate the expressions of 78-kDa glucose-regulated protein (GRP78) and Caspase-12 and their relationship with apoptosis in renal cortex of diabetic rats.

METHODSUninephrectomized Wistar rats were used to induce diabetes by intraperitoneal injection of Streptozotocin (STZ 65 mg/kg). After 8 weeks, the expression and distribution of GRP78, Caspase-12, proliferating cell nuclear antigen (PCNA) were examined by immunohistochemistry. Flow cytometry was used to detect the levels of protein of GRP78 and Caspase-12. Apoptosis was evaluated by means of terminal deoxynucleotidyl transferase-mediated d-UDP nick-end labeling (TUNEL) and Flow cytometry. Serum creatinine, blood urea nitrogen and 24-hour urine protein excretion were checked.

RESULTSCompared with those in normal control group, the numbers of apoptosis and the expression of GRP78, Caspase-12 in glomerular and tubular cells were much higher in the diabetic kidneys at 8 weeks. There was no significant difference between group A and group B.

CONCLUSIONActivation of endoplasmic reticulum stress may play an important role in the development of diabetic nephropathy.

Animals ; Apoptosis ; Caspase 12 ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; pathology ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Kidney Cortex ; metabolism ; pathology ; Male ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Wistar

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

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

Based on the complete genome sequence of pigeon-origin Newcastle disease virus strain JS/07/04/ Pi(genotype VIb), nine overlapped fragments covering its full-length genome were amplified by RT-PCR. The fragments were connected sequentially and then inserted into the transcription vector TVT7/R resulting in the TVT/071204 which contained the full genome of strain JS/07/04/Pi. The TVT/071204 was co-transfected with three helper plasmids pCI-NP, pCI-P and pCI-L into the BSR cells, and the transfected cells and culture supernatant were inoculated into 9-day-old SPF embryonated eggs 60 h post-transfection. The HA and HI tests were conducted following the death of embryonated eggs. The results showed that the allantoic fluids obtained were HA positive and the HA could be inhibited by anti-NDV serum which indicated that the strain JS/07/04/Pi was rescued successfully. The rescued virus rNDV/071204 showed similar growth kinetics to its parental virus in CEF. The successful recovery of this strain would contribute to the understanding of the host-specificity of pigeon-origin NDV and to the development of the novel vaccines against the NDV infection in pigeons.
Animals ; Base Sequence ; CHO Cells ; Chick Embryo ; Columbidae ; virology ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Fluorescent Antibody Technique, Indirect ; Molecular Sequence Data ; Newcastle disease virus ; genetics ; growth & development

OBJECTIVETo understand the current epidemiological characteristics as well as the trends of stomach cancer deaths in China.

METHODSThe data of stomach cancer mortalities in 2004 - 2005, from 158 sampling areas in the Third National Retrospective Sampling Survey of Death Causes in China, were analyzed and compared with the results from previous two national surveys.

RESULTSThe crude and age-standardized death rates of stomach cancer were 24.71/100 000 (35 250/142 660 482) and 16.16/100 000, respectively, accounted for 18.19% (35 250/193 841) and ranking third of cancer causes in the national sampling areas of China in 2004 - 2005. Those crude death rate increased by 42.01% while the age-standardized death rate decreased by 8.70% compared to the results in 1973 - 1975 (17.40/100 000 and 17.70/100 000), and both decreased 1.79% and 25.74% from 1990 - 1992 (25.16/100 000 and 21.76/100 000), respectively. For urban residents of the sampling areas, the crude and age-standardized death rates of stomach cancer were 22.98/100 000 (11 005/47 899 806) and 13.63/100 000, accounted for 15.03% (11 005/71 936) of cancer causes in 2004 - 2005, which increased by 18.21% and decreased by 31.16% from 1973 - 1975 (19.44/100 000 and 19.80/100 000), and increased by 18.21% and decreased by 11.15% from 1990 - 1992 (19.44/100 000 and 15.34/100 000), respectively. While for rural residents in the sampling areas, the crude and age-standardized death rates were 25.59/100 000 (24 245/94 760 676) and 17.64/100 000, accounted for 19.89% (24 245/121 905) of cancer causes, both increased by 53.97% and 3.76% from 1973 - 1975 (16.62/100 000 and 17.00/100 000), and both decreased by 5.78% and 27.59% from 1990 - 1992 (27.16/100 000 and 24.36/100 000), respectively.

CONCLUSIONThe current stomach cancer is still one of predominant cancers in China. The consistently substantial decreases in age-standardized death rates of stomach cancer might prompt the beneficial impact on reducing the risks for that cancer by the social economical development during recent decades in China.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Data Collection ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Retrospective Studies ; Sampling Studies ; Stomach Neoplasms ; epidemiology ; mortality ; Young Adult