A pan-KRAS degrader for the treatment of KRAS-mutant cancers.

KRAS mutations are highly prevalent in a wide range of lethal cancers, and these mutant forms of KRAS play a crucial role in driving cancer progression and conferring resistance to treatment. While there have been advancements in the development of small molecules to target specific KRAS mutants, the presence of undruggable mutants and the emergence of secondary mutations continue to pose challenges in the clinical treatment of KRAS-mutant cancers. In this study, we developed a novel molecular tool called tumor-targeting KRAS degrader (TKD) that effectively targets a wide range of KRAS mutants. TKD is composed of a KRAS-binding nanobody, a cell-penetrating peptide selectively targeting cancer cells, and a lysosome-binding motif. Our data revealed that TKD selectively binds to KRAS in cancer cells and effectively induces KRAS degradation via a lysosome-dependent process. Functionally, TKD suppresses tumor growth with no obvious side effects and enhances the antitumor effects of PD-1 antibody and cetuximab. This study not only provides a strategy for developing drugs targeting "undruggable" proteins but also reveals that TKD is a promising therapeutic for treating KRAS-mutant cancers.

Potent human monoclonal antibodies targeting Epstein-Barr virus gp42 reveal vulnerable sites for virus infection.

Epstein-Barr virus (EBV) is linked to various malignancies and autoimmune diseases, posing a significant global health challenge due to the lack of specific treatments or vaccines. Despite its crucial role in EBV infection in B cells, the mechanisms of the glycoprotein gp42 remain elusive. In this study, we construct an antibody phage library from 100 EBV-positive individuals, leading to the identification of two human monoclonal antibodies, 2B7 and 2C1. These antibodies effectively neutralize EBV infection in vitro and in vivo while preserving gp42's interaction with the human leukocyte antigen class II (HLA-II) receptor. Structural analysis unveils their distinct binding epitopes on gp42, different from the HLA-II binding site. Furthermore, both 2B7 and 2C1 demonstrate potent neutralization of EBV infection in HLA-II-positive epithelial cells, expanding our understanding of gp42's role. Overall, this study introduces two human anti-gp42 antibodies with potential implications for developing EBV vaccines targeting gp42 epitopes, addressing a critical gap in EBV research.

Prolactin receptor potentiates chemotherapy through miRNAs-induced G6PD/TKT inhibition in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive malignancies, with a notoriously dismal prognosis. As a competitive inhibitor of DNA synthesis, gemcitabine is the cornerstone drug for treating PDAC at all stages. The therapeutic effect of gemcitabine, however, is often hindered by drug resistance, and the underlying mechanisms remain largely unknown. It is unclear whether their response to chemotherapeutics is regulated by endocrine regulators, despite the association between PDAC risk and endocrine deregulation. Here, we show that prolactin receptor (PRLR) synergizes with gemcitabine in both in vitro and in vivo treatment of PDAC. Interestingly, PRLR promotes the expression of miR-4763-3p and miR-3663-5p, two novel miRNAs whose functions are unknown. Furthermore, the analysis of transcriptome sequencing data of tumors from lactating mouse models enriches the PPP pathway, a multifunctional metabolic pathway. In addition to providing energy, the PPP pathway mainly provides a variety of raw materials for anabolism. We demonstrate that two key enzymes of the pentose phosphate pathway (PPP), G6PD and TKT, are directly targeted by miR-4763-3p and miR-3663-5p. Notably, miR-4763-3p and miR-3663-5p diminish the nucleotide synthesis of the PPP pathway, thereby increasing gemcitabine sensitivity. As a result, PRLR harnesses these two miRNAs to suppress PPP and nucleotide synthesis, subsequently elevating the gemcitabine sensitivity of PDAC cells. Also, PDAC tissues and tumors from LSL-KrasG12D/+, LSL-Trp53R172H/+, and PDX1-cre (KPC) mice exhibit downregulation of PRLR. Bisulfite sequencing of PDAC tissues revealed that PRLR downregulation is due to epigenetic methylation. In this study, we show for the first time that the endocrine receptor PRLR improves the effects of gemcitabine by boosting two new miRNAs that block the PPP pathway and nucleotide synthesis by inhibiting two essential enzymes concurrently. The PRLR-miRNAs-PPP axis may serve as a possible therapeutic target to supplement chemotherapy advantages in PDAC.

Progesterone receptor potentiates macropinocytosis through CDC42 in pancreatic ductal adenocarcinoma.

Endocrine receptors play an essential role in tumor metabolic reprogramming and represent a promising therapeutic avenue in pancreatic ductal adenocarcinoma (PDAC). PDAC is characterized by a nutrient-deprived microenvironment. To meet their ascendant energy demands, cancer cells can internalize extracellular proteins via macropinocytosis. However, the roles of endocrine receptors in macropinocytosis are not clear. In this study, we found that progesterone receptor (PGR), a steroid-responsive nuclear receptor, is highly expressed in PDAC tissues obtained from both patients and transgenic LSL-KrasG12D/+; LSL-Trp53R172H/+; PDX1-cre (KPC) mice. Moreover, PGR knockdown restrained PDAC cell survival and tumor growth both in vitro and in vivo. Genetic and pharmacological PGR inhibition resulted in a marked attenuation of macropinocytosis in PDAC cells and subcutaneous tumor models, indicating the involvement of this receptor in macropinocytosis regulation. Mechanistically, PGR upregulated CDC42, a critical regulator in macropinocytosis, through PGR-mediated transcriptional activation. These data deepen the understanding of how the endocrine system influences tumor progression via a non-classical pathway and provide a novel therapeutic option for patients with PDAC.

[Effects and Mechanisms of Homoharringtonine on Expression of CEBPA Protein].

OBJECTIVE: To investigate the effect of homoharringtonine (HHT) on CEBPA protein and explore the mechanism of HHT in the treatment of acute myeloid leukemia (AML) with double CEBPA mutations. METHODS: The K562 cell line expressing CEBPA p30 (K562 CEBPA p30) was established. Western blot was used to determine the changes of the expression of CEBPA protein in K562 CEBPA p30, U937 and MOLM-13 cell lines before and after treatments with HHT, daunorubicin (DNR) or cytarabine (Ara-C). The effects of protease inhibitors and protein synthesis inhibitors on the expression of CEBPA protein were also determined. RNA-seq was used to analyze the difference of gene expressions and pathway enrichments between HHT group and DNR group. RESULTS: Both the endogenous CEBPA protein in U937 and MOLM-13 cell lines and the exogenous CEBPA protein in K562 CEBPA p30 were decreased by HHT (P<0.05) while were not by DNR or Ara-C. Proteasome inhibitors can increase the expression of CEBPA protein (P<0.05) while protein synthesis inhibitors can decrease the expression of CEBPA protein (P<0.05). The ribosome biogenesis related pathways in K562 CEBPA p30 were upregulated in HHT group while were not in DNR group. CONCLUSION: HHT can inhibit the synthesis of CEBPA and reduce the expression of CEBPA protein and this may be the mechanism of HHT in the treatment of CEBPA-double-mutant AML.

Meroterpenoids with a large conjugated system from Ganoderma lucidum and their inhibitory activities against renal fibrosis.

Baoslingzhines A-E (1-5), five new meroterpenoids were isolated from the fruiting bodies of Ganoderma lucidum. The structures including their absolute configurations were characterized by using spectroscopic and computational methods. Compound 1 is a novel trinormeroterpenoid featuring the presence of an unusual dihydronaphthalene representing an unprecedented meroterpenoid skeleton. Compounds 2-4 are mononormeroterpenoids characteristic of a large conjugated system. Among them, racemic 3 and 4 were separated by HPLC on chiral phase. Biological evaluation toward kidney fibrosis found that compounds 2 and (+)-3 could inhibit the expression of fibronectin and collagen I dose dependently in TGF-ß1-induced rat kidney proximal tubular cells (NRK-52e). Additionally, (+)-3 could also down regulate ɑ-SMA in a concentration dependent manner. Further investigation showed that 2 could inhibit Smad2 phosphorylation.

Characterization of Ictalurid herpesvirus 1 Glycoprotein ORF59 and Its Potential Role on Virus Entry into the Host Cells.

The channel catfish virus (CCV, Ictalurid herpesvirus 1) has caused sustained economic losses in the fish industry because of its strong infectivity and pathogenicity. Thus, it is necessary to determine the function of viral proteins in the CCV infection process. The present study aimed to characterize CCV glycoprotein ORF59 and explore its impact on virus infection in host cells. Firstly, its exclusive presence in the membrane fraction of the cell lysate and subcellular localization verified that CCV ORF59 is a viral membrane protein expressed at late-stage infection. A protein blocking assay using purified His6 tagged ORF59, expressed in sf9 insect cells using a baculovirus expression system, indicated a dose-dependent inhibitory effect of recombinant ORF59 protein on virus invasion. Knockdown of the ORF59 using a short hairpin (shRNA) showed that ORF59 silencing decreased the production of infectious virus particles in channel catfish ovary cells. The results of this study suggest that recombinant ORF59 protein might inhibit CCV entry into the host cells. These findings will promote future studies of the key functions of glycoprotein ORF59 during CCV infection.

[Research advances in species diversity of arbuscular mycorrhizal fungi in terrestrial agro-ecosystem]. / é™†åœ°å†œä¸šç”Ÿæ€ç³»ç»Ÿä¸›æžèŒæ ¹çœŸèŒç‰©ç§å¤šæ ·æ€§ç ”ç©¶è¿›å±•.

Arbuscular mycorrhizal fungi (AMF) are ancient and ubiquitous soil microorganisms, which can form mutually beneficial association with most terrestrial plants. Within the symbiotic relationship, AMF helps their host plants to absorb nutrients such as nitrogen and phosphorus while obtains carbon from the hosts. AMF plays an important role in agricultural ecosystem, including promoting plant growth, improving crop quality, increasing plant stress resistance, stabilizing soil structure, keeping ecological balance, and maintaining a sustainable agricultural development. We summarized the research advances of AMF in terrestrial agro-ecosystem in recent years, by focusing on AMF species diversity, spatial and temporal distribution, and influence factors of AMF biodiversity in terrestrial agro-ecosystem of China. Further research works were also prospected.

Characterization of a new endo-type polyM-specific alginate lyase from Pseudomonas sp.

An alginate lyase gene, algA, encoding a new poly ß-D-mannuronate (polyM)-specific alginate lyase AlgA, was cloned from Pseudomonas sp. E03. The recombinant AlgA with (His)6-tag, consisting of 364 amino acids (40.4 kDa),was purified using Ni-NTA Sepharose. The purified lyase had maximal activity (222 EU/mg) at pH 8 and 30 °C and also maintained activity between pH 7-9 and below 45 °C. It exclusively and endolytically depolymerized polyM by ß-elimination into oligosaccharides with degrees of polymerization (DP) of 2-5. Due to its high substrate specificity, AlgA could be a valuable tool for production of polyM oligosaccharides with low DP and for determining the fine structure of alginate.

[A matched nested case-control study on the risk factors of metabolic syndrome among male criminal policemen].

OBJECTIVE: To explore the risk factors and their differences of metabolic syndrome (MS) on male criminal police, thereby provide the scientific basis to make prevention and control strategies about the metabolic syndrome for the criminal police career. METHODS: Based on physical examination data of criminal police in 2010, 439 patients with MS (CDS) were randomly selected as cases. And as the 1:2 matched nested case-control study, 878 health controls were employed, which were matched with on the basis of sex and age (±1 year). An face-to-face epidemiological investigations on the past exposure status of several possible risk factors was conducted, such as the family history of hypertension and other social economic status, as well as body height and weight, waist circumference, blood pressure, serum lipid and plasma sugar. and the data were analyzed with logistic regression. RESULTS: 1317 cases were surveyed, through single factor logistic regression analysis found that 12 factors are related to exposure. Multivariate logistic regression analysis suggested that six factors, such as stress events (OR = 1.989, 95%CI: 1.467∼2.696), snoring (OR = 1.672, 95%CI: 1.218∼2.294), sweets (OR = 0.562, 95%CI: 0.412∼0.766), meat and products (OR = 1.494, 95%CI: 1.065∼2.094), siting after dinner for more than 3 h (OR = 1.399, 95%CI: 1.023∼1.915). CONCLUSIONS: MS has become a important public health problems among criminal police. For their professional special, a series of bad habits , unhealthy life style and psychological problems became important risk factors of MS on criminal police. Targeted prevention and control measures should be taken to reduce the incidence of MS.

Identification of a potential hydrophobic peptide binding site in the C-terminal arm of trigger factor.

Trigger factor (TF) is the first chaperone to interact with nascent chains and facilitate their folding in bacteria. Escherichia coli TF is 432 residues in length and contains three domains with distinct structural and functional properties. The N-terminal domain of TF is important for ribosome binding, and the M-domain carries the PPIase activity. However, the function of the C-terminal domain remains unclear, and the residues or regions directly involved in substrate binding have not yet been identified. Here, a hydrophobic probe, bis-ANS, was used to characterize potential substrate-binding regions. Results showed that bis-ANS binds TF with a 1:1 stoichiometry and a K(d) of 16 microM, and it can be covalently incorporated into TF by UV-light irradiation. A single bis-ANS-labeled peptide was obtained by tryptic digestion and identified by MALDI-TOF mass spectrometry as Asn391-Lys392. In silico docking analysis identified a single potential binding site for bis-ANS on the TF molecule, which is adjacent to this dipeptide and lies in the pocket formed by the C-terminal arms. The bis-ANS-labeled TF completely lost the ability to assist GAPDH or lysozyme refolding and showed increased protection toward cleavage by alpha-chymotrypsin, suggesting blocking of hydrophobic residues. The C-terminal truncation mutant TF389 also showed no chaperone activity and could not bind bis-ANS. These results suggest that bis-ANS binding may mimic binding of a substrate peptide and that the C-terminal region of TF plays an important role in hydrophobic binding and chaperone function.

[Role of 16 slices spiral CT angiography with 3-dimensional CT and CT virtual endoscopy in detecting blood supply of lung carcinoma].

BACKGROUND & OBJECTIVE: Angiography, a common method in evaluating blood supply of lung carcinoma, is invasive and complicated, with low success rate for bronchial artery, and could not assure to show all supply blood vessels at a time. This study was to explore clinical value of 16 slices spiral CT angiography with 3-dimensional CT (3DCT) and CT virtual endoscopy (CTVE) in diagnosing and evaluating supply blood vessels and blood supply of lung carcinoma, so as to find a non-invasive, safe, simple and effective method in diagnosing blood supply of lung carcinoma. METHODS: A total of 72 patients with pathologically proved lung carcinoma underwent 16 slices spiral CT angiography with 3DCT. Volume rendering (VR), maximum intensity projection (MIP), and surface shaded display (SSD) of supply blood vessels of lung carcinoma were used as 3DCT models. CTVE of bronchial artery was performed in 25 patients. Color VR of tumor lesion was performed in all patients. RESULTS: Supply blood vessels were showed in 68 patients, 59 of them showed only bronchial artery, 5 showed intercostals arteries, and 4 showed mixed types, including bronchial artery, intercostals arteries, or branch arteries of subclavian artery. The bronchial artery entered into enlarged mediastinal lymph nodes in 4 patients. CTVE well displayed the orifice and lumen of bronchial arteries in the 25 patients. The extent of red color of tumor lesion on VR color image were divided into 4 types: no color (n=11), light red (n=17), moderate red (n=32), and heavy red (n=12); the added CT values of tumor lesion after enhanced CT were (6.16+/-2.23) Hu, (15.71+/-3.13) Hu, (25.47+/-2.71) Hu, and (44.31+/-19.68) Hu, respectively. The corresponding rate between enhanced type and distributive type of red color on color VR was 86.1%. CONCLUSIONS: The 16 slices spiral CT angiography with 3DCT and CTVE could show clearly supply blood vessels and blood supply of lung carcinoma. It is a non-invasive, simple and effective method in evaluating and diagnosing blood supply of lung carcinoma.

[The value of three-phase pulmonary helical CT in diagnosing peripheral pulmonary cancer (diameter

OBJECTIVE: To evaluate the value of three-phase pulmonary helical CT in diagnosing peripheral pulmonary cancer (diameter

[Helical dual-phase CT scan in evaluating blood supply of primary heptocellular carcinoma after transcatheter hepatic artery chemoembolization with lipiodol].

OBJECTIVE: To evaluate the blood supply of low density viable area of primary heptocellular carcinoma after transcatheter hepatic artery chemoembolization using lipiodol (LP-TACE), by helical dual-phase CT scanning and three dimensional CT (3DCT). METHODS: Thirty-four patients with primary heptocellular carcinoma after LP-TACE were examined by hepatic helical dual-phase CT. 3DCT model of the maximum intensity projection (MIP), surface shaded display (SSD) reconstruction of the hepatic artery and portal vein were simultaneously done in 5 cases. RESULTS: Viable tumor areas of 34 cases of primary heptocellular carcinoma after LP-TACE were divided into four types: peripheral, lateral, central and diffused types. Enhanced tumor vessel or tissue in viable tumor area was found during hepatic dual-phase in 17 cases, during hepatic artery-phase only in 8 and hepatic portal vein-phase only in 3. The viable tumor areas were found to have blood supply from the hepatic vein in 2 cases. The viable tumor area unenhanced during hepatic dual-phase was found in 6 cases. In 5 cases, the relation between the viable tumor area and branches of hepatic artery and portal vein was showed by MIP and SSD of hepatic artery and portal vein. CONCLUSION: Hepatic helical dual-phase CT scan with 3DCT is effective in evaluating the blood supply of viable tumor areas and the therapeutic effect of primary heptocellular carcinoma after LP-TACE.