[The efficacy and side effects of rigosertib combined with chemotherapy in KRAS mutant colorectal cancer mice].

Objective: To investigate the effect of rigosertib (RGS) combined with classic chemotherapy drugs including 5-fluorouracil, oxaliplatin, and irinotecan in colorectal cancer. Methods: Explore the synergy effects of RGS and 5-fluorouracil (5-FU), oxaliplatin (OXA), and irinotecan (IRI) on colorectal cancer by subcutaneously transplanted tumor models of mice. The mice were randomly divided into control group, RGS group, 5-FU group, OXA group, IRI group, 5-FU+ RGS group, OXA+ RGS group and IRI+ RGS group. The synergy effects of RGS and OXA on KRAS mutant colorectal cancer cell lines in vitro was detected by CCK-8. Ki-67 immunohistochemistry and TdT-mediated dUTP nick-end labeling (TUNEL) staining were performed on the mouse tumor tissue sections, and the extracted tumor tissue was analyzed by western blot. The blood samples of mice after chemotherapy and RGS treatment were collected, blood routine and liver and kidney function analysis were conducted, and H&E staining on liver sections was performed to observe the side effects of chemotherapy and RGS. Results: The subcutaneously transplanted tumor models were established successfully in all groups. 55 days after administration, the fold change of tumor size of OXA+ RGS group was 37.019±8.634, which is significantly smaller than 77.571±15.387 of RGS group (P=0.029) and 92.500±13.279 of OXA group (P=0.008). Immunohistochemical staining showed that the Ki-67 index of tumor tissue in control group, OXA group, RGS group and OXA+ RGS group were (100.0±16.8)%, (35.6±11.3)%, (54.5±18.1)% and (15.4±3.9)%, respectively. The Ki-67 index of OXA+ RGS group was significantly lower than that in control group (P=0.014), but there was no significant difference compared to OXA group and RGS group (OXA: P=0.549; RGS: P=0.218). TUNEL fluorescence staining showed that the apoptotic level of OXA+ RGS group was 3.878±0.547, which was significantly higher than 1.515±0.442 of OXA group (P=0.005) and 1.966±0.261 of RGS group (P=0.008). Western blot showed that the expressions of apoptosis related proteins such as cleaved-PARP, cleaved-caspase 3 and cleaved-caspase 8 in the tumor tissues of mice in the OXA+ RGS group were higher than those in control group, OXA group and RGS group. After the mice received RGS combined with chemotherapy drugs, there was no significant effect on liver and kidney function indexes, but the combined use of oxaliplatin and RGS significantly reduced the white blood cells [(0.385±0.215)×10(9)/L vs (5.598±0.605)×10(9)/L, P<0.001] and hemoglobin[(56.000±24.000)g/L vs (153.333±2.231)g/L, P=0.001] of the mice. RGS, chemotherapy combined with RGS and chemotherapy alone did not significantly increase the damage to liver cells. Conclusions: The combination of RGS and oxaliplatin has a stronger anti-tumor effect on KRAS mutant colorectal cancer. RGS single agent will not cause significant bone marrow suppression and hepatorenal injury in mice, but its side effects may increase correspondingly after combined with chemotherapy.

[Diagnosis and treatment experience of 161 cases of pancreatic serous cystic neoplasm in single center and analysis of cause of preoperation misjudgement].

Objective: To analyze the causes of preoperative miscarriage of pancreatic serous cystadenoma (SCN) and find the ways to improve it. Methods: Clinical data of 425 pancreatic cystic neoplasm patients who underwent surgical resection from January 2006 to December 2016 in Department of Pancreatic Surgery in Huashan Hospital were retrospectively analyzed.Excel database was created which covered 128 fields of 7 fields: general information of patients, preoperative blood biochemical indexes, tumor markers, surgical related data, postoperative complications, imaging findings and pathology.One hundred and sixty-one cases of SCN were analyzed in depth, mainly in three aspects: surgical benefit, preoperative imaging diagnostic value and interference factors in preoperative judgement.The classification data were analyzed by χ(2) test and the quantitative data were analyzed by t test.The Logistic regression model was used for multiple factor analysis. Results: Of the 425 PCN cases surgically removed, 161 cases (37.9%) were SCN, the incidence of operative complications was 40.4%(65/161), the hospitalization days was (20.7±12.1)days and the medical cost was (75 267±37 866) yuan.Only 3 of 161 cases of SCN were accurately diagnosed by preoperative imaging methods, 61 cases were diagnosed as "cystic lesions of pancreas" (37.9%) and 52 cases were diagnosed as "pancreatic cystadenoma" (32.3%). SCN was misdiagnosed as MCN(32.3%) and IPMN(28%) before operation.25.5% of them were diagnosed as SCN before operation, but still underwent radical operation.The rate of preoperative imaging diagnosis for identifying SCN was 62.8%.The lack of preoperative endoscopy and the lack of understanding of the image characteristics and biological behavior of SCN were the most important factors affecting the accuracy of preoperative judgment.Statistics found that gender, age, CA125 and tumor location can be used as independent factors contribute to the clinical identification(χ(2)=8.995, P=0.003; χ(2)=10.019, P=0.007; t=3.157, P=0.002; χ(2)=6.790, P=0.009). Logistic analysis showed that women, older than 60 years old, the tumors located in the pancreatic body and tail were the independent factors of SCN classification and diagnosis (OR=0.481, 0.376, 0.577, 0.666, 95% CI: 0.305-0.759, 0.199-0.710, 0.361-0.924, 0.433-1.024, P=0.002, 0.003, 0.022, 0.064). Conclusions: SCN has more benign biological behavior.Although surgical excision is acceptable for clinical safety, the corresponding benefit is very limited.It is possible to improve the rationality of SCN clinical operation decisions to some extent by performing endoscopic examination, imaging doctors to improve the SCN feature recognition and surgeons to enhance the awareness of SCN.

Genetic diversity analysis of sweet kernel apricot in China based on SSR and ISSR markers.

Simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers were used to evaluate genetic diversity among 22 sweet kernel apricot accessions and 12 cultivars in China to provide information on how to improve the utilization of kernel apricot germplasms. The results showed that 10 pairs of SSR primers screened from 40 primer pairs amplified 43 allelic variants, all of which were polymorphic (100%), and 9 ISSR primers selected from 100 primers amplified 67 allelic variants with 50 polymorphic bands (74.63%). There was a relatively distant genetic relationship between the 34 samples, where their genetic similarity coefficient was between 0.62 and 0.99. The UPGMA dendrogram constructed using combined data of the two marker systems separated the genotypes into three main clusters.

Sequence organization and evolutionary dynamics of Brachypodium-specific centromere retrotransposons.

Brachypodium distachyon is a wild annual grass belonging to the Pooideae, more closely related to wheat, barley, and forage grasses than rice and maize. As an experimental model, the completed genome sequence of B. distachyon provides a unique opportunity to study centromere evolution during the speciation of grasses. Centromeric satellite sequences have been identified in B. distachyon, but little is known about centromeric retrotransposons in this species. In the present study, bacterial artificial chromosome (BAC)-fluorescence in situ hybridization was conducted in maize, rice, barley, wheat, and rye using B. distachyon (Bd) centromere-specific BAC clones. Eight Bd centromeric BAC clones gave no detectable fluorescence in situ hybridization (FISH) signals on the chromosomes of rice and maize, and three of them also did not yield any FISH signals in barley, wheat, and rye. In addition, four of five Triticeae centromeric BAC clones did not hybridize to the B. distachyon centromeres, implying certain unique features of Brachypodium centromeres. Analysis of Brachypodium centromeric BAC sequences identified a long terminal repeat (LTR)-centromere retrotransposon of B. distachyon (CRBd1). This element was found in high copy number accounting for 1.6 % of the B. distachyon genome, and is enriched in Brachypodium centromeric regions. CRBd1 accumulated in active centromeres, but was lost from inactive ones. The LTR of CRBd1 appears to be specific to B. distachyon centromeres. These results reveal different evolutionary events of this retrotransposon family across grass species.

Genetic variation of 5 SNPs of MC1R gene in Chinese indigenous sheep breeds.

The purpose of this study was to assess genetic diversity, genetic differentiation.relationship and population structure among 10 Chinese sheep populations using 5 single nucleotide polymorphisms (SNPs). in MC1R gene. The genetic diversity indices suggested that the intra-population variation levels of Chinese Merino and Large-tailed Han,breeds were lowest than Kazakh Fat-Rumped. Chinese sheep breeds have maintained a high intra-population variation levels (95.23%). The genetic differentiation patterns and genetic relationships among Chinese sheep breeds displayed a high consistency with the traditional classification. The cluster trees were constructed by UPMGA method. The results showed that Chinese indigenous sheep populations have distinct genetic differentiation. The inter-population variation levels in Chinese sheep populations indicated three geographically independent domestication events have occurred. The Bayesian cluster analyses also showed a reliable clustering pattern, which revealed three major clusters in.Chinese indigenous sheep populations (Mongolian group, Kazakh group and Tibetan group), except for Duolang and Minxian Black-fur. There were probably caused by different breeding history, geography isolation and different levels of inbreeding. The findings supported the related records in literature, ten sheep populations originated on different time stage from the primogenitor population and communicated genetically with each other in the process of natural and artificial selection, and in different ecological environment. It is concluded that Chinese indigenous sheep have higher genetic variation and diversity, genetic differentiation exist between Chinese sheep populations. The majority breeds are consistent with the geographical distribution and breed characteristic.

MicroRNA-338 in MSCs-derived exosomes inhibits cardiomyocyte apoptosis in myocardial infarction.

OBJECTIVE: Myocardial infarction (MI) is caused by myocardial ischemia and hypoxia, which causes irreversible damage to the myocardium and seriously endangers human health. Exosomes are small, monolayer-structured extracellular vesicles that transport proteins, lipids, mRNAs, and miRNAs between cells. Mesenchymal stem cells (MSCs) can secrete a large number of exosomes and play a role in many pathophysiological processes. The purpose of this paper was to investigate the role of exosomal microRNA-338 (miR-338) in MI and its underlying mechanism of action. MATERIALS AND METHODS: We transfected rat bone marrow-derived MSCs with miR-338 mimic or negative control and extracted exosomes secreted by MSCs. Expression of miR-338 in MSCs, exosomes, and H9c2 cells co-cultured with exosomes was detected by PCR. Then, we treated H9c2 cells with H2O2. We transfected miR-338 inhibitor into H9c2 cells co-cultured with exosomes to further study the function of miR-338. Apoptosis of H9c2 cells were observed by Western blot, flow cytometry, and cell staining. We also established a MI rat model to study the function in vivo and injected exosomes in the myocardium. Seven days later, we used echocardiography to detect the heart function of rats. RESULTS: MiR-338 was upregulated in MSCs transfected with miR-338 mimic, exosomes, and H9c2 cells co-cultured with exosomes. When H9c2 cells were co-cultured with exosomes overexpressing miR-338, the expression of Bax was decreased while the expression of Bcl-2 was increased, and the apoptosis rate was also decreased as shown in flow cytometry, and the amount of caspase3 fluorescence was also decreased. Cardiac function was markedly improved after intramyocardial injection of exosomes overexpressing miR-338 in rats. It was demonstrated using computational tools, Western blot, and Luciferase reporter gene experiments that miR-338 could regulate JNK pathway via targeting MAP3K2. CONCLUSIONS: Exosomal miR-338 can inhibit cardiomyocyte apoptosis and improve cardiac function in rats with myocardial infarction by regulating MAP3K2/JNK signaling pathway.

The correlation between the expression of ADAM17, EGFR and Ki-67 in malignant gliomas.

OBJECTIVE: The aim of this study was to analyze the depolymerization in metalloproteinase (ADAM17), epidermal growth factor receptor (EGFR), the expression of Ki-67 of glioma patients and the correlations with malignancy. PATIENTS AND METHODS: 53 brain glioma samples resected from patients who had surgery from April 2015 to May 2016 at Chinese People's Armed Police Force General Hospital were selected. According to the degree of malignancy: 22 patients were divided into a deterioration group (stage I to II); 31 patients in highly deteriorated group (stage III to IV); 14 brain tissue samples of traumatic decompression from the hospital as control group. The immunohistochemical method was used to detect the expression of ADAM17, EGFR, and Ki-67 in three groups, and the correlation between the expression of ADAM17, EGFR, and Ki-67. Thus, the stages of malignancy were analyzed. RESULTS: ADAM17, EGFR, and Ki-67 had no expression or weak expression in the control group, and increased in the low stage of deterioration group; the differences were statistically significant (p < 0.05). The positive expression rates of ADAM17, EGFR, and KI-67 were significantly higher in the high deterioration group than that in the control group (p < 0.05). Moreover, the analysis showed that the expression of ADAM17, EGFR, and Ki-67 were positively correlated with the stage of malignancy (R = 0.823, p = 0.000; R = 0.804, p = 0.000; R = 0.811, p = 0.000). CONCLUSIONS: The results suggested that there was a significant positive correlation between ADAM17, EGFR, and Ki-67 with the stage of malignancy.

The effect of rapamycin on TGFß1 and MMP1 expression in a rabbit model of urethral stricture.

PURPOSE: To investigate the effect of rapamycin on TGFß1 and MMP1 expression in a rabbit model of urethral stricture. METHODS: Twenty-four adult New Zealand male rabbits underwent an electrocoagulation of the bulbar urethra with a 13Fr pediatric resectoscope. Then rabbits were randomly divided into three groups: (1) normal control group: normal saline (NS), (2) the vehicle control group: dimethyl sulfoxide (DMSO), and (3) the treatment group: effective-dose rapamycin in DMSO (Ra), with 12, 6, and 6 rabbits in each group, respectively. Drugs were given by urethral irrigation daily for 4 weeks. Urethral tissue was harvested for histological and molecular analyses. TGFß1 and MMP1 expression levels were evaluated by real-time quantitative PCR and immunohistochemistry. RESULTS: Ten, six, and six rabbits were evaluated finally in Ra, DMSO, and NS group, respectively. Histological examination revealed the distribution of fibrosis and the degree of collagen deposition in the Ra group were smaller and slighter than the two control groups. Collagen content was significantly less in the Ra group than in the DMSO group (P < 0.001) and the NS group (P < 0.001). qRT-PCR analysis showed a higher expression of MMP1 mRNA in the Ra group than in the DMSO group (P < 0.001) and the NS group (P < 0.001). Immunohistochemistry showed the protein levels of MMP1 in the Ra group were significantly increased when compared with the DMSO group (P < 0.01) and the NS group (P < 0.01). On the other hand, no statistical difference could be found between every two groups in both mRNA and protein levels of TGFß1. CONCLUSIONS: Rapamycin enhances the expression of MMP1 in a rabbit model of urethral stricture, but has no direct effect on the expression of TGFß1.

Benefit from synchronous portal-superior mesenteric vein resection during pancreaticoduodenectomy for cancer: a meta-analysis.

BACKGROUND: Pancreaticoduodenectomy combined with portal-superior mesenteric vein synchronous resection for cancer remains a hot debate topic. The present study used meta-analytical technique to provide update information and an evidence-based evaluation on both the perioperative benefit and long-term survival. METHODS: A meta-analysis was performed to evaluate studies comparing venous resection (VR) versus without venous resection (WVR) groups. 22 retrospective studies including 2890 patients were eligible for an analysis of perioperative morbidity, mortality, and long-term survival. Furthermore, subgroup analysis was made according to histopathology and resection margin status respectively for the purpose of survival assessment. RESULTS: There was no difference in perioperative morbidity, mortality and 1-year, 3-year survival between two groups, but showed differences in median tumor size (P < 0.001), R0 resection rate (P < 0.001), lymph node metastasis (P = 0.03), pancreatic fistula (P = 0.01), and 5-year survival (P = 0.03). In subgroup analysis, patients in venous resection group received R0 resection had a significantly better survival comparing with who received R1 resection both at 2-year (P < 0.001) and 5-year (P = 0.00002). In histopathology subgroup, patients in venous resection groups who had true tumor infiltration had a significantly bad survival comparing with whom only with inflammation pathology. CONCLUSION: Pancreaticoduodenectomy combined with venous resection can achieve equal perioperative morbidity and mortality as standard resection. However, in order to obtain an optimal survival outcome, surgeons should make an R0 resection as far as possible, especially in cases need synchronous venous resection.