Design, synthesis and biological evaluation of novel 2,4-diaminopyrimidine cinnamyl derivatives as inhibitors of FAK with potent anti-gastric cancer activities.

In this study, twenty-one novel 2,4-diaminopyrimidine cinnamyl derivatives as inhibitors targeting FAK were designed and synthesized based on the structure of TAE-226, and the inhibitory effects of these compounds on both the FAK enzyme and three cancer cell lines (MGC-803, HCT-116, and KYSE30) were investigated. Among them, compound 12s displayed potent inhibitory potency on FAK (IC50 = 47 nM), and demonstrated more significant antiproliferative activities in MGC-803, HCT-116 and KYSE30 cells (IC50 values were 0.24, 0.45 and 0.44 µM, respectively) compared to TAE-226. Furthermore, compound 12s significantly inhibited FAK activation leading to the negative regulation of FAK-related signaling pathways such as AKT/mTOR and MAPK signaling pathways. Molecular docking study suggested that compound 12s could well occupy the ATP-binding pocket site of FAK similar to TAE-226. In addition, compound 12s also efficiently inhibited the proliferation, induced apoptosis and cellular senescence in MGC-803 cells. In conclusion, compound 12s emerges a potent FAK inhibitor that could exert potent inhibitory activity against gastric cancer cells.

Design, synthesis and evaluation of novel bis-substituted aromatic amide dithiocarbamate derivatives as colchicine site tubulin polymerization inhibitors with potent anticancer activities.

As the continuation of our work on the development of tubulin inhibitors with potential anticancer activities, novel bis-substituted aromatic amide dithiocarbamate derivatives were designed by contacting bis-substituted aryl scaffolds (potential anti-tubulin fragments) with N-containing heterocycles (potential anti-tubulin fragments) in one hybrid using the anticancer dithioformate unit as the linker. The antiproliferative activity against three digestive tract tumor cells was evaluated and preliminary structure activity relationships were summarized. Among these compounds, compound 20q exhibited most potent antiproliferative activity against MGC-803, HCT-116, Kyse30 and Kyse450 cells with IC50 values of 0.084, 0.227, 0.069 and 0.078 µM, respectively. In further studies, compound 20q was identified as a novel tubulin inhibitor targeting the colchicine binding site. Compound 20q could inhibit the microtubule assembly and disrupt cytoskeleton in Kyse30 and Kyse450 cells. The results of molecular docking suggested that compound 20q could tightly bind into the colchicine binding site of tubulin by hydrogen bonds and hydrophobic interactions. Compound 20q dose-dependently inhibited the cell growth and colony formation, effectively arrested cells at the G2/M phase and induce mitochondrial apoptosis in Kyse30 and Kyse450 cells. In addition, Compound 20q could regulate the expression of G2/M phase and mitochondrial apoptosis related proteins. Collectively, compound 20q was here reported as a novel tubulin inhibitor with potential anticancer activities.

Islet transplantation ameliorates diabetes-induced testicular interstitial fibrosis and is associated with inhibition of TGF-ß1/Smad2 pathway in a rat model of type 1 diabetes.

Islet transplantation (IT) is considered the most effective endocrine replacement therapy for diabetes mellitus (DM). Studies have demonstrated that IT can repair testicular structural injury caused by inflammatory and oxidative stress in a diabetic rat model. However, highly effective exogenous antioxidant and anti-inflammatory drugs can achieve this effect. Testicular interstitial fibrosis caused by long-term hyperglycemia is however difficult to reverse or recover. Thus far, there are no effective drugs that prevent or relieve testicular interstitial fibrosis. Therefore, it is necessary to explore the potential benefit of IT on testicular interstitial fibrosis induced by DM and its underlying molecular mechanisms. In the present study, Wistar rats were used to establish a DM model by intraperitoneal injection of streptozotocin. The diabetic models then underwent IT or received insulin treatment after 12 weeks. IT was more effective than insulin treatment in ameliorating diabetic-induced testicular interstitial fibrosis, Leydig cells apoptosis, testosterone deficiency and poor sperm motility. IT and insulin treatment both significantly inhibited the upregulation of TGF-ß1 and phosphorylated Smad2 in DM, with IT being more effective than insulin. The present study's findings proved that IT effectively protects diabetic-induced testicular interstitial fibrosis probably by inhibiting the TGF-ß1/Smad2 signaling pathway, which offers hope in male patients with DM complicating with testicular interstitial fibrosis.

Target genes regulated by transcription factor E2F1 in small cell lung cancer.

Previously, we have reported that transcription factor E2F1 expression is up-regulated in approximately 95% of small cell lung cancer tissue samples and closely associated with invasion and metastasis, but few studies have investigated specific target genes regulated by E2F1 in this disease. The aim of this study was to clarify the target genes controlled by E2F1 in the small cell lung cancer cell line H1688. The results of chromatin immunoprecipitation sequencing (ChIP-seq) showed that total 5 326 potential target genes were identified, in which 4 700 were structural genes and 626 long non-coding RNAs (lncRNAs). Gene Ontology (GO) and enrichment map analysis results indicated that these target genes were associated with three main functions: (1) cell cycle regulation, (2) chromatin and histone modification, and (3) protein transport. MEME4.7.0 software was used to identify the E2F1 binding DNA motif, and six motifs were discovered for coding genes and lncRNAs. These results clarify the target genes of E2F1, and provide the experimental basis for further exploring the roles of E2F1 in tumorigenesis, development, invasion and metastasis, recurrence, and drug resistance in small cell lung cancer.

[Association of adiponectin gene polymorphisms +45T/G with gestational diabetes mellitus and neonate birth weight].

OBJECTIVE: To determine the association of single-nucleotide polymorphisms (SNPs)+45T/G of adiponectin gene with gestational diabetes mellitus (GDM) and neonate birth weight. METHODS: A total of 264 GDM and 272 pregnant women with normal glucose (NG) were enrolled. DNA was successfully extracted from peripheral blood leucocyte samples. And SNPs+45T/G of adiponectin gene were examined. We also analyzed differences in the genotypic distribution and allelic frequencies in SNP+45T/G of adiponectin gene among GDM group with different neonate birth weights. RESULTS: There was no significant difference between two groups with respects to SNP +45T/G polymorphisms of adiponectin gene (P > 0.05). But significant genotypic difference existed in SNP+45T/G among three GDM groups with different birth weights. Frequencies of T allele were significantly higher in the GDM group with higher birth weight, 68.75% in GDM group with macrosomia and 72.86% in GDM group with 3000-4000 g birth weight (both P < 0.01). And there was no significant difference in SNP +45T/G polymorphisms of adiponectin gene between GDM women and NG group with macrosomia. CONCLUSION: SNP +45T/G polymorphisms of adiponectin gene are not associated with GDM. However, SNP+45T/G polymorphisms of adiponectin gene of GDM women are closely associated with neonate birth weigh. The G/T polymorphisms of SNP+45 of adiponectin are not obviously associated with a higher occurrence of macrosomia in GDM women.

[Analysis of the perinatal outcomes and management of twin-twin transfusion syndrome].

OBJECTIVE: To investigate the perinatal outcomes of twin-twin transfusion syndrome (TTTS) and the management. METHODS: During Nov 1, 2002 to Sep 30, 2005, 24 cases of TTTS in Beijing Obstetrics and Gynecology Hospital were analyzed. The outcomes of them were compared with the pregnancy without TTTS in all twins and in monozygotic twins. The outcomes of the blood-supplying fetus and the blood-recepter were compared. RESULTS: 6.8% cases had TTTS in all twins. The group of TTTS had more maternal, fetal and neonatal complications than twins pregnancy without TTTS: polyhydramnios [37.5% (9/24) vs 2.1% (7/328), P < 0.01], gestational hypertension [20.8% (5/24) vs 7.0% (23/328), P = 0.043], premature labor [66.7% (16/24) vs 36.3% (119/328), P = 0.003], perinatal dead fetus in uterus [18.8% (6/32) vs 1.1% (7/640), P < 0.01], neonatal asphyxia [73.1% (19/26) vs 3.0% (19/632), P < 0.01], the proportion of NICU [88.5% (23/26) vs 23.4% (148/632), P < 0.01], neonatal death [15.4% (4/26) vs 1.7% (11/632), P = 0.002] and the rate of perinatal mortality [31.2% (0/32) vs 2.8% (18/632)]. Compared with the monozygotic twins without TTTS, in TTTS group there were more complications of the mother, the fetus and the neonates: gestational hypertension [20.8% (5/24) vs 9.9% (14/142), P = 0.224], premature labor [66.7% (16/24) vs 49.3% (70/142), P = 0.115], perinatal dead fetus in uterus [18.8% (6/32) vs 0.7% (2/282), P < 0.01], neonatal asphyxia [73.1% (19/26) vs 3.9% (11/280), P < 0.01], the proportion of NICU [88.5% (23/26) vs 29.3% (82/280), P < 0.01], neonatal death [15.4% (4/26) vs 2.1% (6/280), P = 0.006] and the rate of perinatal mortality [31.3% (10/32) vs 3.2% (8/282)]. The perinatal outcomes were better in those cases that the grades of TTTS were below 3 in the first diagnosis. CONCLUSIONS: We should try to diagnose and treat TTTS as early as possible because the outcome is poor.

[Proliferation and differentiation of MC 3T3-E1 cells cultured on nanohydroxyapatite/chitosan composite scaffolds].

with chitosan in situ using a chemical method and a porous structure obtained was then lyophilized. Preosteoblast MC 3T3-E1 the scaffolds was examined after staining it with Wright's stain. Their proliferation was assessed using MTZ assay. After being Abstract Nanohydroxyapatite/chitosan composite scaffolds were fabricated and the proliferation and differentiation of preosteoblast MC 3T3-E1 on them were examined for the assessment of their biocompatibility. Nanohydroxyapatite was combined with chitosan in situ using a chemical method and a porous structure obtained was then lyophilized. Preosteoblast MC 333-E1 cells were inoculated into the porous composite scaffolds and chitosan scaffolds, respectively. The morphology of cells cultured on the scaffolds was examined after staining it with Wright's stain. Their proliferation was assessed using MTT assay. After being cultured in conditioned medium for 30 days, the cells' alkaline phosphatase activities on the scaffolds were studied in situ to compare their differentiation levelabout. Moreover, the alkaline phosphatase activities were assessed with a kit. The expression level of characteristic osteogenic gene was evaluated using Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The results indicated that MC 3T3-E1 cells grown on the composite scaffolds showed a higher proliferation rate and spread better than that on chitosan scaffolds. The alkaline phosphatase stain results showed that the alkaline phosphatase activity of cells on composite scaffolds was significantly higher than that on the chitosan scaffolds. In addition, the quantitative examination of alkaline phosphatase activity indicated that the cells cultured on the composite scaffolds expressed an activity level about 8 times higher than that on chitosan scaffolds. Simultaneously, the osteogenic gene osteopontin (OPN) of cells cultured on composite scaffolds showed a higher expression level than that on chitosan scaffolds. Another osteogenic gene osteocalcin (OC) was expressed in cells cultured on composite scaffolds, whereas it was not detected in cells on chitosan scaffolds. The addition of nanohydroxyapatite in the scaffolds improved not only the proliferation but also the differentiation of preosteoblast cultured on them. The composite scaffolds showed good biocompatibility and bioactivity. These scaffolds would be promising in bone tissue engineering.