The mechanism of anti-hepatocellular carcinoma cell line HepG2 by Chinese medicine Astragalus protein / 药学学报

To detect the inhibitory effect of Astragalus protein on the proliferation of hepatocellular carcinoma cell line HepG2, transcriptomics was used to explore the anti-tumor mechanism of Astragalus protein. The dried roots of Astragalus was precipitated by ammonium sulfate to obtain Huang Qi protein (HQP) with different molecular weights. The effect of HQP on HepG2 and its toxic effect were detected by hemocytometry. Cell necrosis was detected by flow cytometry and Hoechst/propidium iodide (PI) double staining. The necrotic marker protein receptor interacting serine/threonine kinase 1 (RIP1) was determined by Western blot. Transcriptome sequencing was performed on the control group and dosing group RNA, and differential expression genes were analyzed for RNA-seq results. qRT-PCR was used to verified the relative mRNA expression levels of candidate genes. The results showed that the inhibition of HepG2 proliferation was more obvious with the increase of HQP concentration. When the concentration of HQP was 100 μg·mL-1, the necrosis rate increased to 18.78%, and the number of red necrotic cells stained with PI was observed under the microscope. The Western blot results showed an increase in RIP1 protein levels. The results of RNA-seq analysis showed that 26 000 related genes were regulated by HQP, and 979 genes were more regulated. KEGG analysis found that some differentially expressed genes were associated with p53 signaling pathway, and qRT-PCR further verified that the sequencing results were reliable. HQP may cause programmed necrosis of HepG2 cells and may be involved in the p53 signaling pathway.

Research on autophagy induced by two xanthone compounds in HepG2 cells / 中国中药杂志

To investigate the inhibitory effects of two xanthone compounds, 1-hydroxy-2,3,4,8-4 methoxy xanthone(here in after referred to as Fr15) and 1-hydroxy-2,3,4,6-4 methoxy xanthone(here in after referred to as Fr17), on the proliferation of hepatocellular carcinoma cells HepG2, and to further investigate their mechanism in combination with transcriptomics. Cell counting was used to detect the effects of two kinds of xanthone compounds Fr15 and Fr17(0, 0.03, 0.15, 0.3 mmoL·L~(-1)) on the proliferation of HepG2 cells; the effects of the two compounds Fr15 and Fr17 on HepG2 cell cycle were detected by flow cytometry; the changes of autophagosomes count in cells were observed under fluorescence microscope; the expression of autophagy marker proteins autophagy marker proteins SQSTM 1(p62) and microtubule associated protein 1 light chain 3 Ⅰ/Ⅱ(LC3 Ⅰ/Ⅱ) in the cells was detected by Western blot; the differentially expressed genes between the control group and the experimental group were analyzed by RNA-seq transcriptome sequencing; qRT-PCR was used to verify the differentially expressed genes in sequencing. The results showed that compounds Fr15 and Fr17 inhibited the proliferation of HepG2 cells with the increase of drug concentration and time. Flow cytometry showed that compounds Fr15 and Fr17 had little effect on HepG2 cell cycle. Fluorescence microscopy results showed that the number of autophagosomes in cells increased with the increase of drug concentration. Western blot showed that the expression of p62 protein was decreased and the expression of LC3-Ⅱ protein was significantly increased after drug addition. The results of RNA sequencing showed that 26 102 and 52 351 differentially expressed genes were obtained in Fr15 and Fr17 respectively. Analysis of KEGG showed that drug treatment had a great effect on autophagy pathway. qRT-PCR verified that 6 up-regulated genes were related to autophagy, and their trend was consis-tent with sequencing results, where all 6 genes showed an up-regulated trend. Two xanthone compounds Fr15 and Fr17 may inhibit proliferation of HepG2 cells by inducing autophagy.

Testicular artery-sparing microscopic varicocelectomy combined with Qilin Pills for bilateral varicocele-induced oligoasthenospermia / 中华男科学杂志

<p><b>Objective</b>To explore the clinical effect of testicular artery-sparing microscopic varicocelectomy (MV) in combination with Qilin Pills (QL) in the treatment of bilateral varicocele-induced oligoasthenospermia.</p><p><b>METHODS</b>Sixty patients with bilateral varicocele-induced oligoasthenospermia were randomly assigned to receive MV (n=30) or MV+QL (n=30), those in the latter group treated with oral QL for 12 weeks postoperatively. At 4, 8, and 12 weeks after operation, we compared the semen volume, sperm concentration, sperm motility, the levels of serum Inh B, luteinizing hormone (LH) and total testosterone (TT), and the testosterone secretion index (TSI) between the two groups.</p><p><b>RESULTS</b>After surgery, all the patients showed disappearance of varicocele symptoms, remarkably improved semen volume, sperm concentration, sperm motility, serum Inh B and TT levels, TSI, decreased LH and FSH (P<0.01). At 12 weeks after treatment, statistically significant differences were found between the MV and MV+QL groups in Inh B (138.96±22.26 vs 129.54±22.23) ng/L, LH (3.17±0.12 vs 3.59±0.11) IU/L, TT (13.98±3.02 vs 12.68±3.12) nmol/L, and TSI (4.41±0.53 vs 3.53±0.51) nmol/ IU (P<0.05). The pregnancy rate was significantly higher in the MV+QL than in the MV group (73.4% vs 36.6%, P<0.05).</p><p><b>CONCLUSIONS</b>Testicular artery-sparing microscopic varicocelectomy combined with Qilin Pills is an effective strategy for the treatment of bilateral varicocele-induced oligoasthenospermia by significantly improving the semen quality of the patient.</p>

Effects of glial cell-derived neurotrophic factor on SCF protein and antioxidant enzyme activity in the testis of unilateral cryptorchidism rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of glial cell-derived neurotrophic factor (GDNF) on the stem cell factor (SCF), superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in the undescended testis tissue of rats.</p><p><b>METHODS</b>Models of left cryptorchidism were made in 48 healthy male rats weighing (200 +/- 20) g and randomly divided into four groups: model control, GDNF 7 d, GDNF 14 d and GDNF 21 d. The rats in the latter three groups were killed at 7, 14 and 21 days after intravenous injection of GDNF, their left testes harvested for measurement of the activities of SOD and CAT and the content of MDA. The apoptosis index of spermatogenic cells was detected by TUNEL, the histological changes of the testis tissue observed under the light microscope, and the gene and protein expressions of SCF determined by real-time quantitative PCR and Western blotting, respectively.</p><p><b>RESULTS</b>The apoptosis indexes of spermatogenic cells were obviously decreased in the GDNF 7 d, GDNF 14 d and GDNF 21 d groups (8.42 +/- 0.16, 4.45 +/- 0.34 and 7.32 +/- 0.09) as compared with that of the model control (13.5 +/- 0.64), with statistically significant difference between the GDNF 14 d and control groups (P < 0.01). The SCF expression and SOD activity were remarkably increased while the MDA content markedly reduced in the GDNF groups in comparison with those in the model control (P < 0.01).</p><p><b>CONCLUSION</b>GDNF had a protective effect on the spermatogenic function of rat testes with unilateral cryptorchidism. It could enhance the antioxidant enzyme activity of the antioxidant enzyme system, elevate the expression of SCF and thus improve the testicular reproductivity, which were best indicated in the GDNF 14 d group.</p>

The genetic screening of a dominant zebrafish mutant in long-term memory / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To screen the learning and memory mutant from N-ethyl-N-nitrosourea (ENU) mutagenic zebrafish F1, and to get the new model animal to study the mechanism of learning and memory.</p><p><b>METHODS</b>Zebrafish mutant was screened by inhibitory avoidance behavioral test and identified by the expression of gene c-fos with qRT-PCR.</p><p><b>RESULTS</b>We isolated a zebrafish mutant related to learning and memory, fgt. In this fgt zebrafish mutant long-term memory was much lower than that in wild-type when tested at 24 h after training. The 24 h long-term memory in about half of fgt mutant F2 (13/30) were significantly lower than those in wild-type, and the others relatively normal. Compared with the expression in wild-type fishes, the expression of immediate-early genes (IEGs) c-fos in half of fgt mutant F2 (13/30) after exploring in a novel environment increased distinctly from the basal control levels statistically, and the others relatively normal, which were in accordance with the behavioral results.</p><p><b>CONCLUSION</b>The zebrafish mutant fgt is a dominant mutant with defect in long-term memory.</p>

Effects of microwave radiation on extraction of epimedin B from Epimedii Folium / 中草药

Objective: To investigate the influence of microwave radiation on the extraction of epimedin B from Epimedii Folium. Methods: The effects of extraction solvent, ratio of material to liquor, and duration of microwave radiation on the extraction yield of epimedin B were studied by single factor tests. The influence of microwave radiation on the stability of epimedin B was assessed by UV-Vis spectrophotometeric analysis. The impacts of microwave radiation on leaf sample were observed using paraffin section method. Results: The contributions of ethanol concentration and duration of microwave radiation for the extraction yield of epimedin B are significant, whereas ratio of material to liquor is not a significant factor. Microwave radiation could result in the disruptions of leaf tissues and other parts, but it did not affect the stability of epimedin B. Conclusion: Microwave radiation could facilitate the extraction of epimedin B, which might result from the enhancement of the mass transfer of ethanol solution into leaf tissues and dissolution of epimdin B from the matrix.