[Anti-gastric cancer effects of Z Ajoene and its molecular mechanisms].

Objective: To investigate the effects of Z Ajoene on gastric cancer cell MGC-803 and its molecular mechanisms. Methods: The gastric cancer cells MGC-803 were treated with 0, 1, 5, 25 and 125 µmol/L Z Ajoene for 24 h, 48 h and 72 h, each with 3 replicate wells. The proliferation activity of MGC-803 cells was analyzed by MTS method, mitochondrial membrane potential was analyzed after JC-1 staining, nuclear type was observed after Hoechst 33342 staining, cytotoxicity was detected by LDH release method, and the apoptosis level and cell cycle were analyzed with flow cytometry. RT-qPCR and Western blot methods were used to evaluate the expression levels of P53, Caspase-3, RAS, ERK, BCL-2, AKT, mTOR and PI3K genes. At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, 20 per group, and were subcutaneously inoculated with gastric cancer cell MGC-803 in the groin. Two days later, each group was injected with Z Ajoene at the doses of 0, 1, 5, 25 and 125 µmol/L, 0.1 ml/time, and was injected every other day. On the 20th day of the first injection of tumor cells, 10 mice in each group were killed, the tumor tissues were taken out and weighed. The survival period of the remaining mice was recorded and the effects of Z Ajoene on the growth and survival period of gastric cancer in tumor-bearing mice were observed. Results: After Z Ajoene treatment, the proliferation activity of MGC-803 cells was significantly inhibited and the apoptosis rate was significantly increased(P＜0.01). The transcription and expression levels of p53, Caspase-3 and BAX genes were significantly increased, while the transcription and expression levels of RAS, ERK1, BCL-2, AKT, mTOR and PI3K genes were decreased markedly(P＜0.01). The tumor inhibition experiments showed that the growth of the tumor could be inhibited and the survival time of the tumor-bearing animals could be greatly prolonged after Z Ajoene treatment(P＜0.01). Conclusion: Z Ajoene has therapeutic effects on gastric cancer, can inhibit the proliferation of gastric cancer cells and induce them apoptosis by regulating the expression of PI3K-AKT-mTOR and RAS-RAF-MEK-ERK signal pathways.

The construction and application of a cell line resistant to novel subgroup avian leukosis virus (ALV-K) infection.

A novel avian leukosis viruses (ALV) subgroup named ALV-K was recently isolated from Chinese indigenous chickens which is different from the subgroups (A to E and J) that have previously been reported to infect chickens. More and more ALV-K strains have recently been isolated from local breeds of Chinese chickens. However, there are no more effective diagnostic methods for ALV-K other than virus isolation followed by envelope gene sequencing and comparison. Viral infection can be blocked through expression of the viral receptor-binding protein. In this study, we have engineered a cell line, DF-1/K, that expresses ALV-K env protein and thereby confers resistance to ALV-K infection. DF-1/K can be used in combination with the ALV-K susceptible cell line DF-1 as a specific diagnostic tool for ALV-K and provides a good tool for further research into the molecular mechanisms of interaction between ALV-K env protein and the host cell receptor.

[Anti-hepatoma effects of Smac analogue Birinapant and its related molecular mechanism].

OBJECTIVE: To investigate the effects of Birinapant on hepatocellular carcinoma cells and its related molecular mechanisms. METHODS: Human hepatocellular carcinoma cells QGY-7701 were treated with 0, 1, 5, 25 and 125 nmol/L Birinapant for 24, 48 and 72 hours respectively, each experiment 3 wells.The proliferation activity of cells, the apoptosis levels, the cells nuclear type, the mitochondrial membrane potential, the transcription and expression levels of genes and the cytotoxicity of Birinapant were analyzed.At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, with 20 mice in each group.The mice were inguinal injected with QGY-7701 cells, and then subcutaneous injected with Birinapant (concentrations ranging from 0, 1, 5, 25, 125 µg/kg) in each group after two days, once every other day.On 18th day since first Birinapant injection, 10 mice were killed in each group to weigh tumor tissue and survival time was recorded from the remaining 10 mice.The effects of Birinapant on the growth of the tumor and the survival time of tumor-bearing mice were observed. RESULTS: Compared with the negative control (NC) group, the proliferation activity of QGY-7701 was inhibited significantly after Birinapant treatment and the apoptosis levels were increased significantly (P<0.01).The cell mitochondrial membrane potential was decreased and the karyotype was changed (P<0.01).At the same time, the transcription and expression levels of genes cellular inhibitor of apoptosis protein 1(cIAP-1), cellular inhibitor of apoptosis protein 2(cIAP-2), ras, raf, mek and erk were significantly decreased (P<0.01), while the expression levels of caspase-3 and caspase-9 genes were up-regulated (P<0.01).Compared with the model group (MG), the growth of the tumor was inhibited significantly and the survival time of the tumor-bearing mice was prolonged after Birinapant treatment (P<0.01). CONCLUSIONS: Birinapant can inhibit the expression of cIAP-1, cIAP-2 and the proteins of Ras-Raf-MEK-ERK signal pathways, so as to activate the mitochondria mediated endogenous apoptosis pathway.Birinapant shows a certain inhibitory effect on liver cancer.

MicroRNA­144 inhibits migration and proliferation in rectal cancer by downregulating ROCK­1.

Cancer of the colon and rectum are two distinct entities, which require different treatment strategies and separate treatment. MicroRNAs (miRNAs) act as critical regulators of genes involved in several biological processes. Aberrant alterations of miRNAs have been found in several types of cancer, including colon cancer and rectal cancer. Extensive catalogues of downregulated miRNAs have been identified for colon cancer, whereas only limited data are available for rectal cancer. An example of miRNA profiling in a previous study found that miRNA (miR)­144 showed aberrant expression and appeared to be rectal cancer­specific, its expression not being reported in colon cancer. In the present study, the role of miR­144 in rectal cancer was investigated. SW837 and SW1463 cell lines were selected as rectal cell carcinoma cells. Using reverse transcription-quantitative polymerase chain reaction, western blot, BrdU, cell migration and cell viability assays, it was found that the expression levels of miR­144 were significantly reduced in the SW837 and SW1463 cell lines, and the overexpression of miR­144 suppressed rectal cancer cell viability, migration and proliferation. In addition, Rho­associated coiled­coil containing protein kinase 1 (ROCK1) was identified as a target of miR­144 in the rectal cancer cells. The supplementation of ROCK1 markedly restored the cell migration and proliferation, which was inhibited by miR­144. Together, the data of the present study demonstrated that miR­144 acts as a tumor suppressor by targeting ROCK1, and indicates the potential of miR­144 as a novel biomarker and target in the treatment of rectal cancer.

[Community structure and distribution of riparian Bambusa rigida along lower Gongjiang River, China].

The community structure and distribution of secondary riparian Bambusa rigida in lower Gongjiang River were studied by the transect sampling method and reverse age-class addition. The species in tree and shrub layers in the riparian B. rigida community had the strong native trait. Along the river gradient, the associated species in tree and shrub layers were fragmented, and associated with shore highland plants, suggesting that their distribution did not meet the RCC theory of continuous riparian law. Plant species in herb layer was in accordance with the RCC law, and the species abundance in lower reach was the greatest with 29 families, 55 genera, and 70 species. B. rigida was absolutely dominant in the riparian communities and adapted to the regulation of tree density and physiological integration. The proliferation ratio of B. rigida rapidly decreased to become stabilized, and the degree of its clump dispersion pattern gradually increased. The average density of secondary riparian B. rigida was 114-141 bamboo trees per clump, and the community was in the mid- and late succession stage.

Prevalence and risk factors of diabetes in patients with Klinefelter syndrome: a longitudinal observational study.

OBJECTIVE: To evaluate the prevalence and risk factors of diabetes in patients with Klinefelter syndrome. DESIGN: Retrospective longitudinal study. SETTING: Medical college hospital. PATIENT(S): Klinefelter group (n = 39) and idiopathic hypogonadotropic hypogonadism (IHH) group (n = 40). INTERVENTION(S): Testosterone replacement therapy. MAIN OUTCOME MEASURE(S): The metabolic parameters, lipid profiles, and sex hormones were compared before and after T replacement therapy. The median duration of follow-up was 4 years in the Klinefelter group and 5.2 years in the IHH group. RESULT(S): The prevalence of diabetes was 20.5% (8 of 39) in the Klinefelter group and 5% in the IHH group. In the Klinefelter group, the incidence of diabetes was 12.5% in patients with 47,XXY karyotype and 57.1% in patients with other atypical karyotypes, such as 46XY/47XXY chimera. In the Klinefelter group, the average (± SD) age at diagnosis of diabetes was 27.1 ± 4.5 years. Four subjects had diabetes before T therapy, and their blood glucose did not improve after T replacement. One patient had a history of acute pancreatitis. Two other subjects had very high triglyceride levels. During the follow-up, body weight increased more in Klinefelter patients than in IHH patients. CONCLUSION(S): The prevalence of diabetes is higher in Klinefelter patients than in IHH patients, possibly owing to abnormal karyotypes. Other risk factors, such as low T level, high body weight, acute pancreatitis, and high triglyceride levels, may also contribute to the development of diabetes.

Selection of reference genes for gene expression studies in PBMC from Bama miniature pig under heat stress.

Heat stress decreases immune function and increases disease susceptibility in stressed animals, which are important factors for industry and public health. We investigated the molecular mechanisms underlying heat stress by profiling the expression of target genes involved in the cellular response in the blood of Bama miniature pigs (Sus scrofa domestica) over 21 days with the use of quantitative RT-PCR (qRT-PCR). Reliable standards were established for the normalization of qRT-PCR. Six potential reference genes were ranked by their stability using the geNorm and NormFinder programs. Ribosomal protein L4 (RPL4) and TATA-box-binding protein (TBP) ranked as the two most stably expressed genes, except on day 21 when beta-2-microglobulin (B2M) was the most stable. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ribosomal RNA 18S (18SRNA) were discarded as reference genes due to their unstable expression patterns. When B2M and TBP genes were selected as standards in combination, rather than GAPDH, a significant upregulation in Toll-like receptor 2 (TLR2) expression was observed after 21 consecutive days of heat stress. These findings suggest that selection of an ideal reference gene is a key step in comparison of transcription profiles in Bama miniature pigs.

A goose-sourced paramyxovirus isolated from southern China.

We report the isolation and characterization of a paramyxovirus from geese in South China during 1997. The isolate, designated as goose paramyxovirus/QingYuan 1997-1 (GPMV/QY97-1), showed pathogenicity to geese and could agglutinate chicken erythrocytes. Its hemagglutinating activity was inhibited by antiavian paramyxovirus serotype 1 (APMV-1) serum. The F gene of isolate was amplified by reverse transcription polymerase chain reaction, and sequence analysis proved that its sequence conformed to that reported in the literature, encoding an F0 protein of 553 amino acids with 13 cysteine residues and 6 potential glycosylation sites. It also contained multiple basic amino acids at the deduced cleavage site of the fusion protein, which is a typical feature of highly virulent APMV-1 strains. Sequences analysis of the F gene of GPMV/QY97-1 revealed a homology with other APMV-1 isolates, with its identity ranging from 84.1% to 99.9% on a nudeotide basis and from 88.8% to 99.6% on an amino acid basis. Phylogenetic analysis of the APMV-1 isolates showed that this isolate most closely resembled the reference APMV-1 strain GD/1/98/Go, which was originally isolated from geese in 1998.