[Cistanches tubulosa water extract protects the testis against cyclophosphamide-induced injury: Effect and mechanism].

OBJECTIVE: To investigate the protective effect of Cistanche tubulosa water extract (CTWE) against cyclophosphamide (CTX)-induced testis injury (TI) in mice and its action mechanism. METHODS: Thirty male mice were equally randomized into a normal control, a CTX-induced TI model control and a CTWE treatment group. After 7 days of adaptive feeding, the mice in the CTWE treatment group were treated intragastrically with CTWE at 10 g/kg/d, those in the normal control and TI model control groups with the same volume of normal saline qd all for 35 successive days, and those in the TI model control and CTWE treatment groups by intraperitoneal injection of cyclophosphamide at 80 mg/kg/d at 7, 14, 21, 28 and 35 days. Then all the animals were weighed, blood samples collected, and their testes and epididymides harvested for detection of the serum T content, examination of semen quality, measurement of testis weight, observation of histopathological changes in the testis, and determination of the levels of super oxide dismutase (SOD) and malondialdehyde (MDA) in the testis tissue and the mRNA expressions of the genes related to the nuclear factor erythroid-2-related factor (Nrf2) signaling pathway. RESULTS: The mice in the TI model control group, compared with the normal controls, showed significant decreases in the body weight (ï¼»34.13 ± 1.56ï¼½ vs ï¼»47.08 ± 1.98ï¼½ g, P < 0.05), testis weight (ï¼»81.82 ± 10.61ï¼½ vs ï¼»148.50 ± 14.82ï¼½ mg, P < 0.05), sperm concentration (ï¼»32.60 ± 5.29ï¼½ vs ï¼»78.90 ± 7.95ï¼½ ×106/ml, P < 0.05), sperm motility (ï¼»45.20 ± 7.09ï¼½% vs ï¼»86.70 ± 5.64ï¼½%, P < 0.05), serum T content (ï¼»7.49 ± 1.03ï¼½ vs ï¼»15.93 ± 1.36ï¼½ ng/L, P < 0.05), and SOD level (ï¼»152.22 ± 10.66ï¼½ vs ï¼»356.10 ± 30.95ï¼½ U/mg prot, P < 0.05), but remarkable increases in the percentage of morphologically abnormal sperm (MAS) (ï¼»39.30 ± 7.36ï¼½% vs ï¼»14.40 ± 3.53ï¼½ %, P < 0.05) and MDA level (ï¼»54.91 ± 5.12ï¼½ vs ï¼»31.71 ± 3.57ï¼½ nmol/mg prot, P < 0.05). The animals treated with CTWE, in comparison with the TI model controls, exhibited markedly increased body weight (ï¼»40.67 ± 2.13ï¼½ vs ï¼»34.13 ± 1.56ï¼½ g, P<0.05), testis weight (ï¼»121.21 ± 17.38ï¼½ vs ï¼»81.82 ± 10.61ï¼½ mg, P<0.05), sperm concentration (ï¼»58.40 ± 9.94ï¼½ vs ï¼»32.60 ± 5.29ï¼½ ×106/ml, P < 0.05), sperm motility (ï¼»72.30 ± 7.51ï¼½ % vs ï¼»45.20 ± 7.09ï¼½ %, P < 0.05), serum T content (ï¼»10.89 ± 1.07ï¼½ vs ï¼»7.49 ± 1.03ï¼½ ng/L, P < 0.05) and SOD level (ï¼»217.69 ± 24.59ï¼½ vs ï¼»152.22 ± 10.66ï¼½ U/mg prot, P < 0.05), but decreased percentage of MAS (ï¼»22.20 ± 6.07ï¼½% vs ï¼»39.30 ± 7.36ï¼½%, P < 0.05) and MDA level (ï¼»36.41 ± 4.27ï¼½ vs ï¼»54.91 ± 5.12ï¼½ nmol/mg prot, P < 0.05). The mRNA expressions of Nrf2, HO-1 and NQO-1 in the testis tissue were significantly down-regulated in the TI model controls compared with those in the normal controls (P < 0.05), and remarkably up-regulated in the CTWE treatment group in comparison with those in the TI model group (P < 0.05), while that of Caspase3 markedly increased in the TI model controls (P< 0.05) and decreased in the CTWE treatment group (P < 0.05). Histopathologically, the testis tissue of the TI model controls showed indistinct outlines from the base of the seminiferous tubule to the lumen surface, with disarranged and reduced layers of spermatogenic cells and decreased number of sperm in the seminiferous tubules, while the structure of the spermatogenic tubules recovered almost to normal in the CTWE treatment group. CONCLUSION: Cistanches tubulosa water extract can effectively inhibit cyclophosphamide-induced testis injury by enhancing the activity of antioxidant enzyme and regulating the expressions of the Nrf2 signaling pathway-related genes.

Identification of Prognostic Related Hub Genes in Clear-Cell Renal Cell Carcinoma via Bioinformatical Analysis.

Objective To identify new genes that correlate with prognosis of clear-cell renal cell carcinoma (ccRCC) via bioinformatics analysis. Methods The gene expression profiles of 62 ccRCC and 54 normal kidney tissues were available from the Gene Expression Omnibus database: GSE12606, GSE36895 and GSE66272. The differentially expressed genes were screened with GEO2R and J Venn online tools. Functional annotation including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) was applied to identify the possible function of the hub genes involved in prognosis of ccRCC. In protein protein interaction network (PPI network), the STRING online tool was used to visualize the network of the differentially expressed genes, and the core gene was selected by MCODE App in Cytoscape software. Finally, GEPIA Survival Plot was performed to assess genes associated with worse survival. Results We totally found 648 differentially expressed genes, including 222 up-regulated genes and 426 down-regulated genes. PPI network showed that in 28 up-regulated genes 7 (CCNE2, CDK1, CDC6, CCNB2, BUB1, TTK and PTTG1) enriched in cell cycle and 4 genes (CCNE2, CDK1, CCNB2 and RRM2) enriched in p53 signaling pathway. GEPIA Survival Plot assay revealed that ccRCC patients carrying CDK1, CCNB2, RRM2, BUB1, and PTTG1 had a worse survival. GEPIA Box Plot showed that BUB1, CCNB2, PTTG1, and RRM2 were over expressed in the ccRCC tissues in contrast to the normal tissues (P<0.05). Conclusion ccRCC patients with the four up-regulated differentially expressed genes including BUB1,CCNB2,PTTG1, and RRM2might manifest a poor prognosis.

[Gene expression profiling of testicular seminoma via bioinformatical analysis].

OBJECTIVE: To investigate the differentially expressed genes related to the survival of testicular seminoma patients and the pathogenesis of the malignancy. METHODS: We obtained the gene expression profiling of testicular seminoma data set GSE8607 from the GEO database and differentially expressed genes in testicular seminoma using the GEO2R online tool. We performed GO and KEGG pathway enrichment via the DAVID website, screened testicular seminoma-related core genes using the STRING and Cytoscape software, and analyzed the survival rate and expression level associated with the core genes through the Kaplan Meier plotter and GEPIA websites. RESULTS: Totally, 591 differentially expressed genes were identified, of which 216 were up-regulated and mainly enriched in 23 biological processes, 8 cell components, 5 biological functions and 7 KEGG pathways, and the other 375 down-regulated and enriched only in the biological process, involving the formation and development of spermatozoa. Thirty-two core genes were obtained via PPI network analysis, including C1QB, CCL2, CCR2, GPR183, ELL, LAPTM5, NPY5R, CD28 and MMP9, which were closely related to the survival rate (P < 0.05), C1QB, CCL2, CCR2, SELL, LAPTM5, CD28 and MMP9, which were highly expressed in the testicular seminoma tissue (P < 0.05), and NPY5R, which was lowly expressed in the testicular seminoma tissue (P < 0.05). CONCLUSIONS: The 8 differentially expressed genes identified in the testicular seminoma tissue contribute to effective prediction of the survival rate of the patients as well as to the diagnosis, prevention and treatment of the disease and development of new anti-cancer drugs.

[Regulatory effect of Jinkui Shenqi Pills on the gene expression of the Nrf2 signaling pathway in cyclophosphamide-induced testis injury in mice].

OBJECTIVE: To investigate the protective effect of Jinkui Shenqi Pills (JSP) against cyclophosphamide-induced testis injury (TI) and its anti-oxidation mechanism in mice. METHODS: Thirty male mice were equally divided into a blank control, a TI model control and a JSP treatment group. The mice in the JSP treatment group were treated intragastrically with JSP and the blank controls with normal saline at 1.2 g/kg qd for 7 days, and then the animals in both the TI model control and JSP treatment groups were injected intraperitoneally with cyclophosphamide at 50 mg/kg, once a week, for 35 days, to induce testis injury. After modeling, all the mice were weighed and sacrificed, followed by detection of the serum T content, measurement of the testis weight, examination of semen parameters in the caudad epididymis, and determination of the levels of super oxide dismutase (SOD) and malondialdehyde (MDA) in the testis tissue and the expressions of relevant genes by qRT-PCR. RESULTS: The mice of the TI model control group, compared with the blank controls, showed significant decreases in the body weight (ï¼»34.63 ± 1.92ï¼½ vs ï¼»48.32 ± 1.64ï¼½ g, P<0.05), testis weight (ï¼»80.00 ± 3.90ï¼½ vs ï¼»140.00 ± 6.10ï¼½ mg, P<0.05), testicular organ coefficient (ï¼»0.22 ± 0.01ï¼½ vs ï¼»0.31 ±0.03ï¼½%, P<0.05), sperm motility (ï¼»48.66 ± 8.08ï¼½% vs ï¼»89.33 ± 4.04ï¼½%, P<0.05), sperm concentration (ï¼»28.42 ± 5.26ï¼½ vs ï¼»77.67 ± 8.73ï¼½ ×106/ml, P<0.05), and levels of serum T (ï¼»8.75 ± 0.96ï¼½ vs ï¼»21.75 ± 1.71ï¼½ pg/ml, P<0.05) and SOD (ï¼»140.82 ± 10.08ï¼½ vs ï¼»358.52 ± 40.41ï¼½ U/mg prot, P<0.05), but remarkable increases in the sperm deformity rate (ï¼»37.33 ± 2.08ï¼½ vs ï¼»15.33±1.53ï¼½%, P<0.05) and MDA level (ï¼»54.89±6.09ï¼½ vs ï¼»30.21±2.17ï¼½ nmol/ng prot, P<0.05). The mice of the JSP treatment group, in comparison with the TI model controls, exhibited markedly increased body weight (ï¼»39.80±2.89ï¼½ vs ï¼»34.63±1.92ï¼½g, P<0.05), testis weight (ï¼»130.00 ± 11.00ï¼½ vs ï¼»80.00 ± 3.90ï¼½ mg, P<0.05), testicular organ coefficient (ï¼»0.28 ± 0.01ï¼½ vs ï¼»0.22 ± 0.01ï¼½%, P<0.05), sperm motility (ï¼»76.00 ± 5.29ï¼½% vs ï¼»48.66 ± 8.08ï¼½%, P<0.05), sperm concentration (ï¼»56.08 ± 4.29ï¼½ vs ï¼»28.42 ± 5.26ï¼½ ×106/ml, P<0.05), and levels of serum T (ï¼»15.50 ± 1.29ï¼½ vs ï¼»8.75 ± 0.96ï¼½ pg/ml, P<0.05) and SOD (ï¼»206.59 ± 16.38ï¼½ vs ï¼»140.82 ± 10.08ï¼½ U/mg prot, P<0.05), but decreased sperm deformity rate (ï¼»25.01 ± 2.99ï¼½% vs ï¼»37.33 ± 2.08ï¼½%, P<0.05) and MDA level (ï¼»35.84 ± 3.61ï¼½ vs ï¼»54.89 ± 6.09ï¼½ nmol/ng prot, P<0.05). The mRNA expressions of NOQ-1, Nrf2 and HO-1 in the testis tissue were significantly lower and that of Caspase-3 remarkably higher in the TI model control than in the blank control group (P<0.05), while those of Nrf2 and HO-1 significantly higher and that of Caspase-3 markedly lower in the JSP treatment group than in the TI model controls (P<0.05). Histopathological images displayed reduced layers of spermatogenic cells in the seminiferous tubules, complete exfoliation of the spermatogenic cells in some of the tubules and decreased number of sperm cells in the TI model controls, which were all found normal in the JSP treatment group. CONCLUSIONS: Jinkui Shenqi Pills can effectively inhibit cyclophosphamide-induced testis injury, which may be related to its effect of regulating the gene expression of the Nrf2 signaling pathway and enhancing the activity of antioxidant enzymes.

An endophytic sanguinarine-producing fungus from Macleaya cordata, Fusarium proliferatum BLH51.

Fermentation processes using sanguinarine-producing fungi other than Macleaya cordata may be an alternative way to produce sanguinarine (SA), which is a quaternary benzo[c]phenanthridine alkaloid possessing antibacterial, anthelmintic, and anti-inflammatory properties. In this study, a SA-producing endophytic fungus strain BLH51 was isolated from the leaves of M. cordata grown in the Dabie Mountain, China. Strain BLH51 produced SA when grown in potato dextrose liquid medium. The amount of SA produced by this endophytic fungus was quantified to be 178 µg/L by HPLC, substantially lower than that produced by the host tissue. The fungal SA--which was analyzed by thin layer chromatography and high-performance liquid chromatography--was shown to be identical to authentic SA. Strain BLH51 was identified as Fusarium proliferatum based on the morphological characteristics and nuclear ribosomal DNA ITS sequence analysis. To the best of our knowledge, this is the first report concerning the isolation and identification of endophytic SA-producing fungi from the host plant, which further proved that endophytic fungi are valuable reservoirs of bioactive compounds.