OBJECTIVETo investigate the expression of RKIP, p65 and pERK in hepatocellular carcinoma (HCC) and theIr correlation with invasion and metastasis of HCC.

METHODSReverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of RKIP mRNA. The expression levels of RKIP, p65 and pERK proteins in HCC tumor and peritumoral tissues were determined by immunohistochemistry and Western blot analysis. Statistical analysis was performed to determine the relationship between their expression and clinicopathological parameters.

RESULTSRKIP protein expression level (RKIP/actin) was 0.579 ± 0.380 in HCCs, 1.178 ± 0.659 in peritumoral tissues and 1.115 ± 0.442 in normal liver tissues. The pERK protein level was 1.023 ± 0.478, 0.605 ± 0.367 and 0.461 ± 0.293, p65 protein level was 0.83 ± 0.376, 0.63 ± 0.337 and 0.466 ± 0.345, respectively. Immunohistochemistry analysis showed that the RKIP positive rates in HCCs, peritumoral tissues and normal liver tissues, were 22.2%, 86.0%, and 93.8%, positive rates of p65 were 73.6%, 56.0% and 37.5%, positive rates of pERK were 65.3%, 38.0% and 31.3%, respectively. Statistical analysis revealed that there was a significant difference in RKIP protein expression levels (P < 0.05), but no significant difference in RKIP mRNA expression levels (P > 0.05) among HCC tumors, peritumoral tissues and normal liver tissues. The p65-positive and pERK-positive rates were higher in tumor tissues than that in peritumoral tissues and in normal liver tissues (P < 0.05), but RKIP-positive rates were lower in tumor tissues than that in paritumoral tissues and normal liver tissues (P < 0.05). RKIP protein expression levels were significantly lower in HCCs with intrahepatic or lymphatic metastasis than that in without. The RKIP positive rates in moderately and well differentiated HCCs were significantly higher than that in poorly differentiated HCCs. There was a relationship between RKIP and pERK expressions (P = 0.04), but RKIP expression was not correlated with p65 expression in HCCs (P = 0.143).

CONCLUSIONSOur findings indicate that the down-regulation of RKIP expression may serve as a predictive marker for HCC development, progression and metastasis, which may contribute to the elevated ERK activity. The inhibiting effect of RKIP on invasion and metastasis of liver cancer cells may be due to the down-regulation of pERK expression rather than p65 expression.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Down-Regulation ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Liver ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Phosphatidylethanolamine Binding Protein ; genetics ; metabolism ; Phosphorylation ; RNA, Messenger ; metabolism ; Transcription Factor RelA ; metabolism

OBJECTIVEFor clarifying the situation of the natural infection of rodents having hemorrhagic fever with renal syndrome (HFRS) virus and to type Hantavirus (HV) using molecular technique in Shenzhen city in 2005, and offering guidance for prevention and control of HFRS.

METHODSData on the host animals was collected from the city of Shenzhen. ELISA and indirect immunofluorscent antibody(IFA) test were applied to the specific antibodies against HV in the sera of captured rats. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues of the HV infected rats were inoculated into Meriones unguiculata to isolate HV. The whole viral RNA was extracted from the lung tissues of the HV infected rats and amplified the partial M fragments with RT-nested-PCR, using the HV genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis.

RESULTS472 rodents were captured from Shenzhen in 2005. Surveillance on rats demonstrated 9.96% rats carrying HV (with a density of 8.25%) and the main host was Rattus norvegicus. In the blood samples of rats, anti-HV IgG antibodies were detectable in 56 cases by IFA, and proved to be positive in 76 cases by ELISA. We successfully isolated a HV strain designated as SZ2083 from Rattus norvegicus for the first time in Shenzhen and was identified to SEO type by RT-nested-PCR. Compared with the coding region of the M gene of HV L99 virus strain, the homologies of nucleotide among them were 97%, but the homology was 76% of the SZ2083 with HTN 76-118 virus strain.

CONCLUSIONResults showed the existence of natural epidemic areas of HFRS in Shenzhen city. Based on the results of sequencing, it is possible that the Seoul strain of HV might be the predominant serotype of virus harbored.

Animals ; China ; epidemiology ; Cities ; Data Collection ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Genotype ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; epidemiology ; veterinary ; Rats ; virology ; Rodentia ; virology

OBJECTIVEIn order to investigate Hantavirus (HV) infection of captured rodents and to understand the genotypes and the molecular characteristic of Hantaviruses in Shenzhen.

METHODSThe captured rodents were classified and the density of distribution was calculated. A total of 472 animals were captured, among which Rattus norvegicus was the dominant group. The total viral RNA was extracted from the lung tissues positive with HV antigens by immunofluorescent assay and gene sequence of M fragment was amplified with RT-nested-PCR by using the Hantavirus genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis.

RESULTSThe results of genotype analysis showed that the Hantaviruses taken from twenty-one lung specimens in Rattus norvegicus in Shenzhen city belonged to the Hantavirus type II (SEOV). Results in homology analysis suggested that the homology among twenty-one samples should be rather high with 95.4% of nucleotide sequence identity and they belonged to the same subtype. Phylogenetic tree analysis showed that they were branched into at least six different lineages, and were highly homologized with SZ2083. We also found that these virus strains had not shown more highly homology of nucleotide sequence in nearest district, whereas revealed consistency in farther district.

CONCLUSIONThe major hosts of Hantaviruses in Shenzhen city were Rattus norvegicus and the epidemic strains were genotyped as SEO-type. Nucleotide sequence and deduced amino acid sequence from different rodents were highly homologous, while nucleotide mutation had also been observed. Further studies are required to explore the possible viruses' sequence mutation.

Animals ; China ; epidemiology ; DNA Primers ; DNA, Viral ; Genotype ; Hantavirus ; classification ; genetics ; Hantavirus Infections ; epidemiology ; veterinary ; virology ; Polymerase Chain Reaction ; RNA, Viral ; Rats ; Rodent Diseases ; epidemiology ; virology ; Sequence Homology

Objective Cell cycle-associated protein 1 (Caprin-1) is closely related to the development and progression of cancer. This study aimed to explore the expression of Caprin-1 in the clinical glioma specimen and its influence on the biological char-acteristics of the glioma cell line. Methods Brain tissue specimens were collected from 29 glioma patients and 2 normal humans that died of accidental trauma. A stably transfected U251 cell line with overex-pressed Caprin-1 was established,and the U251 cells were transfected with the pEGFP-C1 plasmid (the negative control group), or the pEGFP-C1-Caprin-1 plasmid (the experimental group), or left un-transfected (the blank control group). The expressions of Caprin-1 mRNA and protein in the cells were determined by RT-PCR and Western blot, and the proliferation and migration of the cells exam-ined by scratch test and Transwell assay,respectively. Results The expression of Caprin-1 was upregulated with the increased grade of glioma,145.9±22.0,444.4±110.0,and 1661.0±54.5 in WHO gradeⅡ,Ⅲ,andⅣglioma,respectively,significantly higher than in the normal brain tissue (P<0.05). Both the mRNA and protein expressions of Caprin-1 were remarkably higher in the experimental group (1.70±0.19 and 1.07±0.09) than in the blank control(0.89±0.10 and 0.52±0.04) and negative control(0.98±0.08 and 0.58± 0.03) (P<0.05).The A value was also markedly higher in the former group(2.55±0.14) than in the latter two(1.40±0.06 and 1.35± 0.04) (P<0.01),and so were the count of migrated cells(526.00±42.19 vs 289.00±29.24 and 279.00±32.48,P<0.01) and the ex-pression of CyclinD1 (0.60±0.05 vs 0.13±0.03 and 0.15±0.05, P<0.01). Conclusion The expression of Caprin-1 in the U251 cells was upregulated with the increased WHO grade of glioma,and the overexpression of Caprin-1 accelerated the proliferation and mi-gration of the U251 cells.

OBJECTIVE: To estimate the minimal clinically important difference (MCID) of the frequency of bowel movement for the patients with chronic severe functional constipation treated with acupuncture so as to provide the evidence for the clinical decision. METHODS: In this study, 813 patients with chronic severe functional constipation treated with acupuncture in two previous randomized controlled trials were included. Through the anchor-based method (anchored by the item 28 "satisfaction with previous treatment" of the patient assessment of constipation-quality of life [PAC-QOL]) and the distribution-based method, the MCID of the weekly frequency of complete spontaneous bowel movement (CSBM) and spontaneous bowel movement (SBM) was analyzed statistically in the patients. RESULTS: The MCID of the mean weekly frequency of CSBM and SBM was 1.3 times and 1.6 times in patients with chronic severe functional constipation treated with acupuncture, respectively. CONCLUSION The mean increase of the weekly CSMB is ≥ 1.3 times and that of SBM is ≥ 1.6 times after treatment when compared with the baseline respectively, suggesting the clinical significance.
Humans ; Quality of Life ; Minimal Clinically Important Difference ; Treatment Outcome ; Constipation/therapy* ; Acupuncture Therapy