Screening and Identification of ESR1 as a Target of Icaritin in Hepatocellular Carcinoma: Evidence from Bibliometrics and Bioinformatic Analysis.

BACKGROUND: In 2022, icaritin a Traditional Chinese Medicine with estrogen-like activities was recommended by the CSCO guidelines as a systematic treatment for patients with advanced HCC due to its clinical safety and efficacy. However the mechanism and targets of icaritin are unclear. In this study we aimed to reveal the target of icaritin in HCC. METHODS: First literature related to icaritin was downloaded from the Web of Science. The software programs "Rstudio" "VOSviewer" and "Mendeley Desktop" were used to analyze the distribution of icaritin publications and research hotspots. Meanwhile icaritin-related genes were obtained by combining them with the PubChem database. Second transcriptome data of HCC patients were obtained from the TCGA database. The proteinprotein interaction (PPI) analysis of icaritin-related genes was performed using the String data platform and the visualization and network topology analysis were performed using Cytoscape. Cox regression analyses were combined to screen the hub target and verified it through cell experiments. RESULTS: A total of 239 icaritin-related articles were obtained HCC is a new hotspot in the icaritin field. 292 icaritin-related genes were obtained and a core module containing 34 genes was obtained by module division. Among them ESR1 was an independent prognostic factor. Molecular docking showed that ESR1 and icaritin had a high affinity. Functional studies revealed that ESR1 inhibits HCC cell malignant proliferation and improves the sensitivity of HCC cells to icaritin. CONCLUSION: We propose that ESR1 as a target of icaritin may be conducive to improving icaritin therapy.

[Autologous CIK cell infusion promotes amplification ability of CD3⁺ CD56⁺ cells when re-preparation from patients with malignant tumors].

OBJECTIVE: To investigate the effects of the autologous cytokine-induced killer (CIK) cell infusion on the subpopulation distribution and activity of the CIK cells from the patients with malignant tumors when prepared in the same liquid culture system again. METHODS: A total of 201 patients who gave a written consent and received 2 courses of therapeutic infusion of CIK cells were divided into ≤ 90-day group in which the secondary preparation of CIK cells was performed in less than 90 days after the primary infusion and >90-day group in which the secondary preparation of CIK cells was performed in more than 90 days after the primary infusion. The proliferation and subtypes, including CD3⁺ cells, CD3⁺ CD4⁺ cells, CD3⁺ CD8⁺ cells, and CD3⁺ CD56⁺ cells, of CIK cells were analyzed by hemocytometer with trypan blue exclusion and flow cytometry, respectively. The expression of NKG2D receptor was also detected using flow cytometry. The cytotoxicity against K562 cells was analyzed using lactate dehydrogenase (LDH) release. RESULTS: The percentage of CD3⁺ CD56⁺ cell subpopulation in the secondary preparation of CIK cells in ≤ 90-day group [(16.7 ± 9.1)%] was higher than that in the primary preparation of CIK cells [(13.5 ± 8.6)%] (P<0.01). Furthermore, the percentage of NKG2D in the secondary CIK cell preparation [(84.1 ± 10.8)%] was significantly higher than that in the primary CIK cell preparation [(81.1 ± 14.8)%] in ≤ 90-day group (P<0.05). In contrast, the percentage of CD3⁺ CD4⁺ cells in the secondary CIK cell preparation [(15.2 ± 9.7)%] was significantly lower than that in the primary CIK cell preparation [(17.6 ± 12.5)%] (P<0.01). However, no significant differences in CD3⁺ CD56⁺ cell subpopulation and expression of NKG2D was detected between the primary and secondary CIK cell preparation in >90 d group, although the percentage of CD3⁺ CD4⁺ cells in the secondary CIK cell preparation [(14.5 ± 9.4)%] was significantly lower than that in the primary CIK cell preparation [(18.2 ± 12.9)%] (P<0.01). In addition, no significant differences in total cell number and cytotoxic activity against K562 cells between the primary and secondary CIK cell preparation was detected either in >90-day group or in ≤ 90-day group. CONCLUSION: CIK cell infusion can facilitate and enhance the proliferation and differentiation of the precursor cells of the CD3⁺ CD56⁺ subpopulation in the same CIK cell culture system and this effect does not last more than 90 days, suggesting that the secondary CIK cell infusion should be performed within 90 days in order to obtain the better therapeutic efficacy.

Characterization of an organic solvent-tolerant lipase from Idiomarina sp. W33 and its application for biodiesel production using Jatropha oil.

A halophilic strain W33 showing lipolytic activity was isolated from the saline soil of Yuncheng Salt Lake, China. Biochemical and physiological characterization along with 16S rRNA gene sequence analysis placed the isolate in the genus Idiomarina. The extracellular lipase was purified to homogeneity by 75% ammonium sulphate precipitation, DEAE-Sepharose anion exchange and Sephacryl S-200 gel filtration chromatography. The molecular mass of the purified lipase was estimated to be 67 kDa by SDS-PAGE. Substrate specificity test indicated that it preferred long-chain p-nitrophenyl esters. Optimal lipase activity was found to be at 60 °C, pH 7.0-9.0 and 10% NaCl, and it was highly active and stable over broad temperature (30-90 °C), pH (7.0-11.0) and NaCl concentration (0-25%) ranges, showing excellent thermostable, alkali-stable and halotolerant properties. Significant inhibition by diethyl pyrocarbonate and phenylarsine oxide was observed, implying histidine and cysteine residues were essential for enzyme catalysis. In addition, the lipase displayed high stability and activity in the presence of hydrophobic organic solvents with log P(ow) ≥ 2.13. The free and immobilized lipases produced by Idiomarina sp. W33 were applied for biodiesel production using Jatropha oil, and about 84 and 91% of yields were achieved, respectively. This study formed the basic trials conducted to test the feasibility of using lipases from halophile for biodiesel production.

[Comparison of effect of formulas clearing away heat and promoting blood circulation on prevention and treatment of liver fibrosis in CCl4 mice].

OBJECTIVE: To observe the effect of traditional Chinese medicine formulas clearing away heat and promoting blood circulation-Biejiayinzi (BJYZ), Gexiazhuyu Tang (GXZYT) and Fugan Wan (FGW) on liver fibrosis in CCl4 mice by screening and analyzing formula-syndrome database of kidney and liver fibrosis based on the principle of formula-syndrome, compared with pivot-harmonizing decoction. METHOD: Ten-week-old male C57BL/6 mice, with the weight of (20 +/- 3) g, were randomly divided into 6 groups: the normal group, the model group, the BJYZ group, the GXZY group, the FGW group and the XST group. Except the normal group, other groups were abdominally injected with 10% CCl4 olive oil solution a dose of 2 mL x kg(-1) body weight for four weeks, three times each week. Meanwhile, the latter four groups were administered with extracts of BJYZ, GXZYT, FGW and XST, respectively, once every day, concomitantly continued CCl4 administration. The normal and the model groups were given the same volume of deionized water. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (gamma-GT), Alb and TBil were detected by chemiluminescence. The hydroxyproline (HYP) content was detected by acid hydrolysis method. The hepatic collagen deposition was evaluated with Sirius red staining. RESULT: Compared with the normal group, the model group recorded notable decrease in weight and increase in the ratio of liver weight and body weight and the ratio of spleen weight and body weight, with obvious fatty degeneration and inflammatory necrosis in liver cells. Collagen fiber deposition was so notable to form fibrous septums and pseudolobules. The levels of serum ALT, AST, TBil, gamma-GT, the HYP content in liver tissue and the deposition of hepatic collagen were significantly increased in the model group. Compared with model group, Serum AST were significantly decreased in BJYZ group as gamma-GT decreased in the GXZYT group, without notable decrease in degeneration and inflammatory necrosis in liver cells and collagen deposition. The GXZYT group showed significant decrease in gamma-GT, with slight improvement in degeneration and inflammatory necrosis in liver cells and reduction in collagen deposition. The ratio of liver weight and body weight, AST, gamma-GT and HYP content were significantly decreased in the FGW group, with slight improvement in degeneration and inflammatory necrosis in liver cells and reduction in collagen deposition. The XST group showed decrease in the ratio of liver weight and body weight, with no obvious change in inflammation and fibrosis of hepatic tissue. CONCLUSION: FGW shows the best effect of prevention and treatment of liver fibrosis in CCl4 mice.