Objective To investigate the effect of ginsenoside Rg1 on the apoptosis of spermatogenic cell in rats with experimentally induced varicocele.Methods A total of 32 adolescent male Wistar rats were included and randomly divided into 4 groups, including sham operation, left varicocele induced, left varicocele induced and low-dose ginsenoside Rg1(5 mg/kg), left varicocele induced and high-dose ginsenoside Rg1 (10 mg/kg), 8 rats in each group.The experimentally induced varicocele was established by Turner method.4 weeks after modeling, the two treatment groups received ginsenoside Rg1 of different dosages by gavage and sham operation and left varicocele induced group received the same volume of NS by gavage, once daily.4 weeks after gavaging, the rats were sacrificed then tissue from left testicle were taken, and the spermatogenic cell apoptosis was detected by TUNEL and Bcl-2 and caspase-3 expressions were detected by immunohistochemistry.Results The protein level of Bcl-2 in ginsenoside Rg1 intervention group was significantly higher than varicocele group, and it was obviously higher in high-dose ginsenoside Rg1 group than low-dose ginsenoside Rg1 group (P<0.05).However, the expression of caspase-3 in ginsenoside Rg1 intervention group was significantly lower than varicocele group, but was higher than sham operation, the expression of caspase-3 in high dose ginsenoside Rg1 group was obviously lower than low dose ginsenoside Rg1 group (P<0.05).The apoptotic index was lowest in sham operation, and which was significantly lower in ginsenoside Rg1 intervention group than varicocele group, and the apoptotic index in high-dose ginsenoside Rg1 group was obviously lower than low-dose ginsenoside Rg1 group ( P<0.05 ) . Conclusion Ginsenoside Rg1 could significantly reduce the testicular sperm cells apoptosis in experimental varicocele rats, raise the Bcl-2 gene expression and inhibiting the expression of caspase-3.

Adrenal Cortex Hormones ; Antibodies, Monoclonal, Murine-Derived ; Humans ; Rituximab ; Thrombocytopenia ; gamma-Globulins

Hypericin, used as a medicinal heifo since ancient times, is a natural photosensitizer derived from the plant Hypericum perforantum. Hypericin would produce peroxides to induce apoptosis and inhibit the growth of oncocytes. Meanwhile, hypericin has a specific affinity with tumor pathology organization (underlying accumulation in pathology organization) which has been widely used in optical diagnosis. The present review gives a comprehensive summary of the anticancer effect based on photodynamic therapy, and photodynamic diagnosis of hypericin optical properties, with the purpose of making full use of the hypericin natural resources, and providing a basis for research and development of hypericin derivatives. © 2006 Editorial office of Foreign Medical Sciences.

Objective: To study the chemical constituents of solid fementation product of endophyte Myrothecium roridum LLY from Ephedra equisetina and their in vitro cytotoxic activity against cancer cells. Methods: Isolation and purification were performed through silica gel and Sephadex LH-20 gel filtration column chromatography as well as HPLC method, and structures of obtained compounds were elucidated by HR-MS and NMR comparised with the data of literatures. In vitro cytotoxicity was evaluated through MTT method and preliminary cytotoxicity mechanism was studied using flow cytometry and Western blotting. Results: Nine compounds were isolated from the solid fementation product of endophyte M. roridum, which were identified as 1-phenazinecarboxylic acid (1), 1-methoxyphenazine (2), N-methyl-1H-indole-2-carboxamaide (3), monomethylsulochrin (4), ditryptophenaline (5), lumichrome (6), deoxyleporin B (7), uracil (8), and thymine (9), respectively. In vitro cytotoxicity assay showed that compound 3 was cytotoxic to the human hepatoma cell line SMMC-7721 with an IC50 value of 38.0 μg/mL, while positive control cisplatin had an IC50 value of 11.5 μg/mL. Cell cycle analysis indicated that compound 3 could delay SMMC-7721 cells in S phase and Western blotting showed that compound 3 could up-regulate the expression of cell cycle negative regulation protein p27 and induce the cleavage of PARP. Conclusion: Compound 7 is a new natural product. Compound 3 could induce the apoptosis of SMMC-7721 cells and the anti-proliferation activity of compound 3 to SMMC-7721 cells is related to S phase arrested and regulated by protein p27.

The aim of this study was to construct the eukaryotic expression vector of pCMV-Myc-NOVA1 based on NOVA1 gene, and to screen the optimum expression condition after transfecting to PC12 cells, and further to explore the distribution of NOVA1 protein in PC12 cells using cell immunohistochemistry, and to identifyits anti-hypoxia activity. According to the NOVA1 gene sequence of NCBI database, we designed the upstream and downstream primers, and performed polymerase chain reaction (PCR) to amplify the full length cDNA coding sequence using pCR4-TOPO-NOVA1 as a template. The products were digested by restriction endonuclease SalⅠand XhoⅠ, and conjugated to the eukaryotic expression vector ofpCMV-Myc followed by validating by digestion and direct sequencing. Subsequently, the validated pCMV-Myc-NOVA1 was transfected to PC12 cells followed by optimizing of transfection ratio and transfection time, and identified by qPCR, Western blotting and cell immunohistochemistry respectively. After validation by digestion and direct sequencing, the eukaryotic expression vector of pCMV-Myc-NOVA1 was correctly constructed. The optimum transfection ratio of plasmid to Lipo 2000 was 1:2.5, and the optimum transfection time was 72 h. At the optimum transfection condition, the expression level of NOVA1 mRNA and protein significantly increased, and after transfection of pCMV-Myc-NOVA1, NOVA1 protein mainly distributed in cell nucleus and cytoplasm. After 6 h hypoxia, the cell proliferation activity was significantly increased compared to that of the control and pCMV-Myc group. Our findings provided a reference for exploring the mechanism of NOVA1, and also a technical support for potential drug development of NOVA1.
Animals ; Blotting, Western ; Cell Hypoxia ; Genetic Vectors ; Humans ; PC12 Cells ; Plasmids ; RNA, Messenger ; RNA-Binding Proteins ; genetics ; metabolism ; Rats ; Transfection

Autoimmune liver disease is a refractory disease clinically, and there is no particularly effective drug at present. Therefore, it is of important clinical value to develop new effective intervention drugs for the prevention and treatment of autoimmune liver disease. In order to investigate the potential protective effect of artesunate (Art) on concanavalin A (Con A)-induced autoimmune liver injury, different doses of Art (27, 54, 108 mg·kg⁻¹) were orally administered to mice for consecutive 7 days, respectively. Then the Con A was injected into mice via tail vein to induce liver injury models. 8 h after modeling, the mice were sacrificed. The serum and liver tissue were collected for detecting the level of alanine aminotransferase (ALT), and aspartate transaminase (AST), liver pathological histopathology, inflammatory cytokines and nuclear factor (NF-κB) key protein expression level. The results showed that 108 mg·kg⁻¹ Art remarkably reduced Con A-induced liver indexes and serum transaminase levels (ALT and AST) as compared with model group(<0.01). Meanwhile, the liver histopathological changes were obviously alleviated with a significant decrease of pro-inflammatory cytokine including tumor necrosis factor (TNF-α), interferon (IFN-γ), interleukin (IL-6), IL-17 and a higher increase of anti-inflammatory cytokine IL-10 (<0.01). Western blot results showed that 108 mg·kg⁻¹ Art markedly inhibited the expressions of p-p65 and p-IκBα proteins (<0.01). The specific inhibitor of NF-κB, pyrrolidinedithiocarbamate (PDTC) could also significantly inhibit the expressions of p-p65 and p-IκBα with and alleviate liver injuries. Therefore, our results indicated that Art may have a protective action against Con A-induced autoimmune liver injury mainly by suppressing NF-κB signal pathway in mice. The study provides scientific reference for artesunate usage in preventing autoimmune liver injury.

In this study, the mobilomes of nine teleost species were annotated by bioinformatics methods. Both of the mobilome size and constitute displayed a significant difference in 9 species of teleost fishes. The species of mobilome content ranking from high to low were zebrafish, medaka, tilapia, coelacanth, platyfish, cod, stickleback, tetradon and fugu. Mobilome content and genome size were positively correlated. The DNA transposons displayed higher diversity and larger variation in teleost (0.50% to 38.37%), was a major determinant of differences in teleost mobilomes, and hAT and Tc/Mariner superfamily were the major DNA transposons in teleost. RNA transposons also exhibited high diversity in teleost, LINE transposons accounted for 0.53% to 5.75% teleost genomic sequences, and 14 superfamilies were detected. L1, L2, RTE and Rex retrotransposons obtained significant amplification. While LTR displayed low amplification in most teleost with less than 2% of genome coverages, except in zebrafish and stickleback, where LTR reachs 5.58% and 2.51% of genome coverages respectively. And 6 LTR superfamilies (Copia, DIRS, ERV, Gypsy, Ngaro and Pao) were detected in the teleost, and Gypsy exhibits obvious amplication among them. While the SINE represents the weakest ampification types in teleost, only within zebrafish and coelacanth, it represents 3.28% and 5.64% of genome coverages, in the other 7 teleost, it occupies less than 1% of genomes, and tRNA, 5S and MIR families of SINE have a certain degree of amplification in some teleosts. This study shows that the teleost display high diversity and large variation of mobilome, there is a strong correlation with the size variations of genomes and mobilome contents in teleost, mobilome is an important factor in determining the teleost genome size.

Estrogen receptor (esr) mediates the effects of estrogen on the expression of related genes, thereby regulating the growth and reproduction of mammals. To investigate the effect of retrotransposon insertion polymorphism (RIP) of the porcine esr gene on porcine growth performance, retrotransposon insertion polymorphism of the esr gene were predicted by comparative genomics and bioinformatics, and PCR was used to verify the insertion polymorphisms in different porcine breeds. Finally, the correlation analysis between the genotypes and performance of Large White pigs was conducted. The results showed that four retrotransposon polymorphic sites were identified in the esr1 and esr2 genes, which are esr1-SINE- RIP1 located in intron 2 of the esr1 gene, esr1-LINE-RIP2 and RIP3-esr1- SINE located in intron 5 of the gene, and esr2-LINE-RIP located in intron 1 of the esr2 gene, respectively. Among them, insertion of a 287 bp of SINE into intron 2 of the esr1 gene significantly affected (P<0.05) the live back fat thickness and 100 kg body weight back fat thickness of Large White pigs. Moreover, the live back fat thickness and back fat thickness at 100 kg body weight of homozygous with insertion (SINE+/+) was significantly greater than that of heterozygous with insertion (SINE+/-) and homozygous without insertion (SINE-/-). Therefore, esr1-SINE-RIP1 could be used as a molecular marker to assist the selection of deposition traits in Large White pigs.
Animals ; Genotype ; Introns/genetics* ; Phenotype ; Polymorphism, Genetic/genetics* ; Retroelements/genetics* ; Swine/genetics*

Systemic lupus erythematosus (SLE) is an autoimmune disease with causes including activation of innate and adaptive immune systems. SLE patients are with a high risk of thrombosis, which may be due to disease activation, immune complexes, toxic antibodies and high level of inflammation. This article discusses neutrophil/NET factor, antibody factor, platelet factor and particle factor which is involved in coagulation pathways and thrombus formation mechanism under the state of immune disorders in SLE.
Blood Coagulation ; Blood Coagulation Factors ; Humans ; Lupus Erythematosus, Systemic ; immunology ; Thrombophilia ; immunology ; Thrombosis

Isolation and purification of chemical constituents of liquid culture of symbiotic Chaetomium globosum ML-4 of oyster was performed through silica gel column chromatography, gel filtration over Sephadex LH-20, preparative TLC and HPLC. Five compounds were obtained and their structures were determined as chaetoglobosin V(1), chaetoglobosin Vb(2), tyrosol(3), 5-methyluracil(4)and uracil(5), respectively, based on HR-MS and NMR data and comparison with literatures. In vitro cytotoxicity of compounds against human hepatocellular carcinoma cell line SMMC-7721 were measured byMTT method, and results showed that compound 1 could obviously inhibit the proliferation of SMMC-7721 cells with an IC₅₀ value of 60.5 mg•L⁻¹, while the IC₅₀ value of positive control cisplatin was 19.96 mg•L⁻¹. Further studies discovered that compound 1 could lead to G2 phase arrest in SMMC-7721 cells and induce SMMC-7721 cells apoptosis. The ratio of Bcl-2/Bax in SMMC-7721 cells was decreased. The expression of protein Caspases-3,-8,-9 was improved and the expression and phosphorylation level of Akt were reduced. Aforementioned results revealed that in vitro antitumor activity of compound 1 against SMMC-7721 cells were related to G2 phase cell cycle arrest and induced-apoptosis. The induced-apoptosis was involved in both the mitochondrial pathway and the death receptor pathway and connected with activity decline of PI3K/Akt signaling pathway.