ObjectiveTo evaluate the effect of interventional chemotherapy (ICT)on the expression of vascular endothelial growth factor(VEGF) and microvessel density (MVD) in rectal carcinoma.MethodsThe expression of VEGF and MVD in 34 patients of rectal cancer were determined before the ICT and 4～5 weeks after the ICT. ResultsBefore the ICT and 4～5 week after the ICT, the expressions of VEGF in the tissue of rectal cancer was 61.8%(21/34) and 32.3%(11/34) respectively(P

Cullin-RING E3 ligases (CRLs) are the major components of ubiquitin-proteasome system, responsible for ubiquitylation and subsequent degradation of thousands of cellular proteins. CRLs play vital roles in the regulation of multiple cellular processes, including cell cycle, cell apoptosis, DNA replication, signalling transduction among the others, and are frequently dysregulated in many human cancers. The discovery of specific neddylation inhibitors, represented by MLN4924, has validated CRLs as promising targets for anti-cancer therapies with a growing market. Recent studies have focused on the discovery of the CRLs inhibitors by a variety of approaches, including high through-put screen, virtual screen or structure-based drug design. The field is, however, still facing the major challenging, since CRLs are a large multi-unit protein family without typical active pockets to facilitate the drug design, and enzymatic activity is mainly dependent on undruggable protein-protein interactions and dynamic conformation changes. Up to now, most reported CRLs inhibitors are aiming at targeting the F-box family proteins (e.g., SKP2, β-TrCP and FBXW7), the substrate recognition subunit of SCF E3 ligases. Other studies reported few small molecule inhibitors targeting the UBE2M-DCN1 interaction, which specifically inhibits CRL3/CRL1 by blocking the cullin neddylation. On the other hand, several CRL activators have been reported, such as plant auxin and immunomodulatory imide drugs, thalidomide. Finally, proteolysis-targeting chimeras (PROTACs) has emerged as a new technology in the field of drug discovery, specifically targeting the undruggable protein-protein interaction. The technique connects the small molecule that selectively binds to a target protein to a CRL E3 via a chemical linker to trigger the degradation of target protein. The PROTAC has become a hotspot in the field of E3-ligase-based anti-cancer drug discovery.
Antineoplastic Agents ; pharmacology ; therapeutic use ; Drug Design ; Drug Discovery ; Enzyme Inhibitors ; pharmacology ; therapeutic use ; Humans ; Neoplasms ; enzymology ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitination ; drug effects

Objective : To study the effect of leucine rich repeat kinase 2 ( LRRK2) on pain sensitivity in neuro- pathic pain (NP) rats and explore its possible mechanism． Methods : 48 SD rats were randomly divided into four groups : sham surgery (Sham) ，model，LRRK2 inhibitor(MLi-2) ，and LRRK2 inhibitor + p38 mitogen activated pro- tein kinase (MAPK) agonist (MLi-2 + Anisomycin) ，with 12 rats in each group．The NP rat model was induced by chronic constriction injury ( CCI) of the sciatic nerve．Intrathecal injection of MLi-2 ( 1 mg / kg，10 μl) or Anisomy- cin (20 μmol / L，10 μl) was started from the 8 th day after surgery，once a day for 7 consecutive days．Pain sensi- tivity tests were conducted before surgery (day 0) and on postoperative days 7 and 14，respectively．The changes in mechanical withdrawal threshold (MWT) and paw withdraw thermal latency (PWTL) were analyzed in each group of rats．ELISA was used to detect the levels of interleukin-1 β (IL-1 β) ，IL-6 and tumor necrosis factor-α (TNF-α) in the dorsal horn of the rat spinal cord．Nissl staining was used to observe the pathological changes of neurons in rat spinal cord tissue．Immunofluorescence staining was used to observe the expression levels of ionized calcium-binding adapter molecule-1 (Iba-1) ，a marker of microglia in the spinal cord of rats ．Western blot was used to detect theprotein expression levels of LRRK2，p-p38 mitogen activated protein kinase (MAPK) ，p38 MAPK，and Iba-1 in the dorsal horn of the rat spinal cord. Results : Compared with the sham group，the model group showed a significant decrease in MWT and PWTL in the right hind limb of rats (P＜0. 01) ．The levels of IL-1，IL-6，and TNF in the spi- nal dorsal horn tissue，as well as the expression levels of LRRK2，Iba-1 proteins and p-p38 MAPK / p38 MAPK pro- tein ratio significantly increased (P＜0. 01) ．The proportion of Iba-1 positive cells in the spinal cord tissue signifi- cantly increased (P＜0. 01) ，while Nissl bodies were significantly reduced (P＜0. 01) ．Compared with the model group，the MLi-2 group showed a significant increase in MWT and PWTL in the right hind limb of rats (P＜0. 01) ， a significant increase in Nissl bodies (P＜0. 01) ，a significant decrease in the proportion of Iba-1 positive cells in the spinal cord tissue (P＜0. 01) ，and a significant decrease in the levels of IL-1，IL-6，and TNF and the expression levels of LRRK2，Iba-1 proteins and p-p38 MAPK / p38 MAPK protein ratio (P＜0. 01) ．However，Anisomychin in- tervention could activate the p38 MAPK signaling pathway and partially reverse the beneficial effects of MLi-2 on pain sensitivity and neuroinflammation in rats with neuropathic pain． Conclusion Inhibiting the expression of LRRK2 can alleviate pain sensitivity in NP rats induced by microglia activation mediated neuroinflammation，and its mechanism of action may be related to regulating the p38 MAPK signaling pathway.

Objective To investigate the role and mechanism of silent information regulator 1 (SIRT1)-NOD-like receptor protein 3 (NLRP3) axis in the effect of remifentanil against ischemia-reperfusion injury (IRI) in rat livers. Methods SD rats were randomly divided into sham operation group (sham group), IRI group, IRI+remifentanil pretreatment group (IRI+RPC group), IRI+SIRT1 inhibitor EX-527 group (IRI+EX-527 group) and IRI+RPC+EX-527 group, with 8 rats in each group. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), interleukin (IL)-1β and IL-18 of rats in each group were detected. The liver tissue pathology was observed. The apoptosis rate of hepatocytes in rats was detected. The expressions of SIRT1, NLRP3, cleaved cysteinyl aspartate specific proteinase-1 (Cleaved Caspase-1) and Gasdermin D (GSDMD) proteins in rat liver tissue were detected. Results Compared with the sham group, the liver tissue pathological score and hepatocyte apoptosis rate of rats in the IRI group were increased, the serum ALT, AST, LDH, IL-1β, and IL-18 levels were increased, the relative expression of SIRT1 protein in liver tissue was decreased, and the relative expression of NLRP3, Cleaved Caspase-1, and GSDMD proteins were increased (all P<0.05). Compared with the IRI group, the liver tissue pathological score and hepatocyte apoptosis rate of rats in the IRI+RPC group were decreased, the serum ALT, AST, LDH, IL-1β, and IL-18 levels were decreased, the relative expression of SIRT1 protein in liver tissue was increased, and the relative expression of NLRP3, Cleaved Caspase-1, and GSDMD proteins were decreased; the liver tissue pathological score and hepatocyte apoptosis rate of rats in the IRI+EX-527 group were increased, the ALT, AST, LDH, IL-1β, and IL-18 levels were increased, the relative expression of SIRT1 protein in liver tissue was decreased, and the relative expression of NLRP3, Cleaved Caspase-1, and GSDMD proteins were increased (all P<0.05). Compared with the IRI+RPC group, the liver tissue pathological score and hepatocyte apoptosis rate in the IRI+RPC+EX-527 group were increased, the levels of ALT, AST, LDH, IL-1β, and IL-18 were increased, the relative expression of SIRT1 protein in liver tissue was decreased, and the relative expression of NLRP3, Cleaved Caspase-1, and GSDMD proteins were increased (all P<0.05). Conclusions SIRT1 may participate in the regulation of remifentanil against rat liver IRI by inhibiting NLRP3 mediated cell pyroptosis.

Protein neddylation is catalyzed by a three-enzyme cascade, namely an E1 NEDD8-activating enzyme (NAE), one of two E2 NEDD8 conjugation enzymes and one of several E3 NEDD8 ligases. The physiological substrates of neddylation are the family members of cullin, the scaffold component of cullin RING ligases (CRLs). Currently, a potent E1 inhibitor, MLN4924, also known as pevonedistat, is in several clinical trials for anti-cancer therapy. Here we report the discovery, through virtual screening and structural modifications, of a small molecule compound HA-1141 that directly binds to NAE in both