Objective To investigate the relationship of replication factor C subunit 2(RFC2)with the prognosis of glioblastoma(GBM)and cell proliferation,as well as its underlying molecular pathway in GBM development.Methods Using bioinformatics methods,cell cycle genes were screened as independent prognostic factors for GBM.Combined with clinical indicators,a risk scoring model for GBM patients was established and validated.The target gene RFC2 was analyzed with GO,KEGG,and GSEA.U87 GBM cells at logarithmic growth stage were transfected with lentivirus and divided into different groups(control,ShRFC2 # 1,and shRFC2 # 2 groups).qRT-PCR,Western blotting,Edu staining,and cloning assay were used to detect mRNA expression,protein expression,and cell proliferation.Results The expression of RFC2 was upregulated in GBM and showed an obvious upregulation trend with the increase of pathological grade of glioma.The analyses of gene function and pathway indicated that RFC2 was involved in the processes of sister chromosome segregation,chromosome segregation,organelle fission,and mitosis by promoting the transition of G1 to S phase during cell cycle.qRT-PCR and Western blotting showed that compared with the control group,the amount of mRNA and translated protein in the knockdowned groups decreased(P＜0.000 1).The positive rate of Edu staining and the colony forming ability decreased(P＜0.000 1,P＜0.001).Conclusion RFC2 is highly expressed in glioblastoma and associated with pathological grade of glioma and poor prognosis of patients.It also promotes the cell proliferation function of glioblastoma.RFC2 may be a potential biomarker and therapeutic target for glioblastoma.

Pheochromocytomas and paragangliomas(PPGLs)cause symptoms by altering the circulation levels of catecholamines and peptide hormones.Currently,the diagnosis of PPGLs relies on diagnostic imaging and the detection of catecholamines.In this study,we used ultra-performance liquid chromatography(UPLC)/quadrupole time-of-flight mass spectrometry(Q-TOF MS)analysis to identify and measure the perioperative differential metabolites in the plasma of adrenal pheochromocytoma patients.We identified differentially expressed genes by comparing the transcriptomic data of pheochromocytoma with the normal adrenal medulla.Through conducting two steps of metabolomics analysis,we identified 111 differential metabolites between the healthy group and the patient group,among which 53 metabolites were validated.By integrating the information of differential metabolites and differentially expressed genes,we inferred that the cysteine-methionine,pyrimidine,and tyrosine metabolism pathways were the three main metabolic pathways altered by the neoplasm.The analysis of transcription levels revealed that the tyrosine and cysteine-methionine metabolism pathways were downregulated in pheochromocytoma,whereas the pyrimidine pathway showed no significant difference.Finally,we developed an optimized diagnostic model of two metabolites,L-dihydroorotic acid and vanylglycol.Our results for these metabolites suggest that they may serve as potential clinical biomarkers and can be used to supplement and improve the diagnosis of pheochromocytoma.

【Objective】 To explore the expressions of adipocyte enhancer binding protein 1 （AEBP1） gene and its isoforms in different types of gliomas， and the influence of AEBP1 gene on the prognosis of patients with different types of gliomas. 【Methods】 We used the GEPIA2 visual network analysis tool to analyze AEBP1 gene expression levels in the tumor tissues of glioblastomas （GBM， including classical， mesenchymal， neural， and proneural ones） and low-grade gliomas （LGG， including astrocytoma， oligoastrocytoma， oligodendroglioma） in the TCGA database and normal human tissue samples in the TCGA and GTEx databases by one-way ANOVA. The distribution trend of isoforms of AEBP1 gene in gliomas was analyzed using the violin plot. The Kaplan-Meier survival curve was drawn and the Logrank test was used to analyze the influence of AEBP1 gene expression in GBM and LGG tumor tissues on the prognosis of glioma patients. 【Results】 The expression of AEBP1 in the tumor tissues of overall GBM and the four types of GBM was higher than that in the normal control tissues （P<0.05）. The expression of AEBP1 in astrocytoma and oligodendrocyte astrocytoma tumor tissues was higher than that in normal control tissues （P<0.05）. There were nine isoforms of AEBP1 gene in GBM and LGG， and the expression level in GBM was higher. The overall survival （OS） of the AEBP1 low expression group of GBM patients and the proneuronal GBM patients was better than that of the high expression group （P<0.05）. The OS and progression-free survival of LGG patients and the AEBP1 low-expression group of astroglioma were better than those of the high-expression group （P<0.05）. 【Conclusion】 AEBP1 has an important clinical value in the pathogenesis and development of GBM and LGG， and thus can be used as a diagnostic marker and a candidate gene for targeted therapy.

Multiple plant lineages have independently evolved sex chromosomes and variable kary-otypes to maintain their sessile lifestyles through constant biological innovation.Morus notabilis,a dioecious mulberry species,has the fewest chromosomes among Morus spp.,but the genetic basis of sex determination and karyotype evolution in this species has not been identified.In this study,three high-quality genome assemblies were generated for Morus spp.[including dioecious M.notabilis(male and female)and Morus yunnanensis(female)]with genome sizes of 301-329 Mb and were grouped into six pseudochromosomes.Using a combination of genomic approaches,we found that the putative ancestral karyotype of Morus species was close to 14 protochromosomes,and that sev-eral chromosome fusion events resulted in descending dysploidy(2n=2x=12).We also charac-terized a～6.2-Mb sex-determining region on chromosome 3.Four potential male-specific genes,a partially duplicated DNA helicase gene(named MSDH)and three Ty3_Gypsy long terminal repeat retrotransposons(named MSTG1/2/3),were identified in the Y-linked area and considered to be strong candidate genes for sex determination or differentiation.Population genomic analysis showed that Guangdong accessions in China were genetically similar to Japanese accessions of mul-berry.In addition,genomic areas containing selective sweeps that distinguish domesticated mul-berry from wild populations in terms of flowering and disease resistance were identified.Our study provides an important genetic resource for sex identification research and molecular breeding in mulberry.

Objective The aim of this study was to investigate the expression of cell cycle checkpoint kinase 1(Chk1)gene in glioblastoma cells( GBM) and its correlation with GBM cell proliferation,tumorigenic activity and prognosis. Methods The ex-pression of Chk1 in GBM cells was selected and analyzed by TCGA database and brain tumor molecular database( Rembrandt),and the level of Chk1 expression in GBM cells was detected by molecular biology techniques such as Western blot and Real-Time PCR. The expression of Chk1 was silenced by siRNA to investigate its effect on proliferation and colony-forming ability of GBM cells. The prognosis survival of GBM patients accompanying with Chk1 expression was analyzed by immunohistochemical staining and Rembrandt database. Results The results of TCGA database and Rembrandt showed that Chk1 gene was highly expressed in GBM tissues. West-ern blot and Real-Time PCR also showed that Chk1 gene was highly expressed in GBM cells. Lentiviral transfection siRNA-specific silencing of Chk1 significantly inhibited proliferation and colony-forming ability of U87 cells( P<0. 01 and P<0. 05). Prognostic survival analysis showed that GBM patients with low expression of Chk1 gene had a significantly better clinical outcome than those of GBM patients with high expression of Chk1 gene(P<0. 001). Conclusion Chk1 gene is overexpressed in GBM cells,up-regula-tion of Chk1 gene expression can promote the growth and proliferation of GBM cells,and Chk1 gene is associated with poor prognosis in GBM patients.

Objective: To discuss the etiology, pathogenesis, clinical manifestation, diagnosis and therapy of sphenoid wing dysplasia(SWD) associated with neurofibromatosis type Ⅰ(NF-Ⅰ). Methods: We retrospectively reviewed its clinical manifestations, imaging, surgical treatment, complications and postoperative outcome of one NF-Ⅰ patient with SWD. Results: A 14 years-old girl presented with pulsating exophthalmos, loss of vision and café au lait spots. Radiological studies showed right-side orbital enlargement and complete absence of the greater wing of the sphenoid. Titanium mesh was tailored intraoperatively to close the defect as a barrier between the orbital cavity and the cranium and then covered by periosteum.The patient developed postoperative infectious which was controlled by after antibiotic treatment and proper drainage. Proptosis improved significantly after surgery within a month. Ocular pulsation subsided and clinical symptoms improved at 28-month follow-up. Conclusions Sphenoid greater wing dysplasia associated with neurofibromatosis type Ⅰ is a rare inherited autosomal dominant disorders. The treatment should be customized to each patient. Titanium mesh reconstruction is patients with symptomatic sphenoid dysplasia. It can correct the proptosis and pulsating exophthalmos without the risk of bone resorption and recurrence.However, high risk of infection is associated with the procedure.

Objective To investigate the clinical effect of cisplatin combined with cisplatin and radiotherapy in the treatment of glioblastomas patients receiving surgery.Methods 68 cases of glioblastomas patients with surgery were divided into control group (n=36) and treatment group (n=32).The control group was treated with temozolomide and radiotherapy;the treatment group was treated with cisplatin combined with temozolomide and radiotherapy.Short-term efficacy,adverse reactions,and survival time were documented during following up in both groups.Results The efficiency in the treatment group (87.5％) was significantly higher than that in the control group (66.67％),P＜0.05.There were no significant difference of adverse reaction including bone marrow inhibition,digestive tract symptoms,and psychiatric symptoms among the two groups,P＞0.05.The medium survival time of the treatment group (26.0 month) was significantly longer than that of the control group (17.5 month),P ＜0.05.There was no significant difference of 1-year survival rate between the treatment group (81.25％) with control group (63.89％),P＞0.05;however,2-and 3-year survival rate in treatment group (52.13％,31.25％) were significantly higher than that in the control group (27.78％,11.11％),P＜0.05.1-year progression free survival (PFS) rate of treatment group (68.75％) was significantly higher that of control group (41.67％),P＜0.05.Conclusion Cisplatin combined with temozolomide and radiotherapy can significantly improve the treatment efficiency without leading to higher rate of adverse reactions and significantly improve the 2-,3-year survival rate as well as 1-year progression free survival (PFS) rate in patients with glioblastomas,it is worth being promoted in clinical application.

ABSTRACT:Objective To investigate the effects of poly ADP-ribose polymerase (PARP ) inhibitor 3-aminobenzene (3-AB)on the delayed development cerebral vasospasm (DCVS)after subarachnoid hemorrhage (SAH)and on the inflammatory factors,namely monocyte chemotactic protein 1 (MCP-1 )and hypersensitive c-reactive protein (hsCRP),and to explore the relationship between these and the signaling pathway of NF-kappa B (NF-κB).Methods Eighty male Sprague-Dawley rats were randomly divided into four groups:normal group (n =8),sham-operation group (n =8),SAH model group (n =32)and 3-AB group (n =32).We established 64 SAH model animals by double injection of blood into the cisterna magna.Half of the SAH model animals were treated with 3-AB by intraperitoneal injection (30 mg/kg).These rats were killed to obtain specimens respectively at days 3, 5,7 and 14 after the second blood injection.The morphological changes of basilar arteries were observed under the light microscope.The contents of PARP,MCP-1 and hsCRP in brain tissues were detected with enzyme-linked immunosorbent assay (ELISA).The expression of NF-κB in basilar arteries was determined by immunohistochemistry.
Results Compared with those in the sham-operation group,the degree of basilar artery spasm reached the peak [(30.47±3.89)%]at day 5 after established SAH model;the thickness and diameter of basilar artery were (1 6.44 ±1.32)μm and (1 78.21 ± 1 1.13)μm,respectively.Cerebral blood flow was reduced by nearly 60% (P <0.01 ). The expression of NF-κB in the cytoplasm and nucleus and PARP content in brain tissue were both increased significantly (P < 0.01 ).MCP-1 [(365.29 ± 28.08 )pg/mL ] and hsCRP [(402.1 6 ± 48.99 )ng/mL ] were significantly enhanced (P <0.01).Compared with the SAH group,after 5 days’intervention with 3-AB,there was obvious alleviation in the spasm degree of basilar artery [(22.65±3.21)%],the thickness [(14.89±1.27)μm]and diameter [(1 98.56±10.91)μm],respectively (P <0.01).Cerebral blood flow was significantly enhanced,but the expression of NF-κB in the cytoplasm and nucleus was decreased and PARP in brain tissue was significantly decreased (P < 0.01 ).MCP-1 [(126.5 1 ± 18.67 )pg/mL]and hsCRP [(285.39 ± 39.07 )ng/mL]in brain tissue were significantly declined,respectively (P <0.01).Conclusion PARP inhibitor 3-AB can alleviate DCVS and inhibit the inflammatory response in brain tissue after SAH.The mechanism may be related to NF-κB signaling pathway.

The previous pharmacokinetic methods can be only limited to drug analysis in vitro, which provide less information on the distribution and metabolismof drugs, and limit the interpretation and assessment of pharmacokinetics, the determination of metabolic principles, and evaluation of treatment effect. The objective of the study was to investigate the pharmacokinetic characteristics of gene recombination angiogenesis inhibitor Kringle 5 in vivo. The SPECT/CT and specific 131I-Kringle 5 marked by Iodogen method were both applied to explore the pharmacokinetic characteristics of 131I-Kringle 5 in vivo, and to investigate the dynamic distributions of 131I-Kringle 5 in target organs. Labeling recombinant angio-genesis inhibitor Kringle 5 using 131I with longer half-life and imaging in vivo using SPECT instead of PET, could overcome the limitations of previous methods. When the doses of 131I-Kringle 5 were 10.0, 7.5 and 5.0 g/kg, respectively, the two-compartment open models can be determined within all the metabolic process in vivo. There were no significant differences in t1/2α, t1/2β, apparent volume of distribution and CL between those three levels. The ratio of AUC(0 ? 1) among three different groups of 10.0, 7.5 and 5.0 g/kg was 2.56:1.44:1.0, which was close to the ratio (2:1.5:1.0). It could be clear that in the range of 5.0–10.0 g/kg, Kringle 5 was characterized by the first-order pharmacokinetics. Approximately 30 min after 131I-Kringle 5 was injected, 131I-Kringle 5 could be observed to concentrate in the heart, kidneys, liver and other organs by means of planar imaging and tomography. After 1 h of being injected, more radionuclide retained in the bladder, but not in intestinal. It could be concluded that 131I-Kringle 5 is mainly excreted through the kidneys. About 2 h after the injection of 131I-Kringle 5, the radionuclide in the heart, kidneys, liver and other organs was gradually reduced, while more radionuclide was concentrated in the bladder. The radionuclide was completely metabolized within 24 h, and the distribution of radioactivity in rats was similar to normal levels. In our study, the specific marker 131I-Kringle 5 and SPECT/CT were suc-cessfully used to explore pharmacokinetic characteristics of Kringle 5 in rats. The study could provide a new evaluation platform of the specific, in vivo and real-time functional imaging and pharmacokinetics for the clinical application of 131I-Kringle 5.

Objective To investigate the effect of supernatant from carcinoembryonic antigen -related cell adhesion molecule 1(CEACAM1)gene RNAi glioma SHG44 cells on human umbilical vein endothelial cell proliferation and angiogenesis in vitro .Methods Three pairs of specific siRNA targeting CEACAM 1 were de-signed and synthesized , and then transiently transfected into SHG 44 cells via cathodolyte liposome transfection method.The supernatant from cultured glioma SHG 44 cells was collected as the conditional medium 48 h after transfection.RT-PCR was used to detect CEACAM1 and VEGF expression at mRNA level after CEACAM1 gene RNAi.The expression of VEGF in supernatant was measured by ELISA .The proliferation of HUVEC was assessed by MTT method after co-cultured with supernatant .The migration of HUVEC was examined by using a Transwell assay.The ability of angiogenesis was measured by capillary -like network formation assay .Results Forty -eight hours after the 3 pairs of specific CEACAM1 siRNA were transfected into SHG44 cells,RT-PCR results showed that the expression of CEACAM 1 mRNA was significantly inhibited compared with those in the normal control and the negative control groups (P<0.01).The most significant interference effect was CEACAM 1-siR-NA3.Expression of VEGF mRNA was remarkably reduced ,and expression of VEGF protein in the supernatant was also reduced after transfection of CEACAM 1-siRNA for 48 h.The proliferation of HUVEC was inhibited , HUVEC migration and tube capillary -like formation were decreased .Conclusion CEACAM1-siRNA can ef-fectively suppress the expression of CEACAM 1 mRNA in human glioma SHG44 cells,may inhibit HUVEC prolif-eration,migration and tube capillary -like formation via down -regulated the secretion of VEGF in vitro .