Objective The objective of this study was to investigate the relationship between the expression of RAD18 and radiation resistance in glioblastoma multiforme(GBM)and to provide a new therapeutic target for improving the radiation resistance of GBM.Methods Human glioma A172 cells were transfected into blank and RAD18-containing plasmid vector.The cell proliferation of two groups after the same dose radiation was detected by cloning assay.The mRNA expression of RAD18 in primary and recurrent GBM samples after close proximity treatment were detected by qRT-PCR.All data were analyzed statistically.Results The proliferation of GBM cells transfected with RAD18 plasmid was higher than that of cells transfected with blank plasmid after radiation therapy(P<0.001).The expression level of RAD18 mRNA in recurrent GBM was higher than that in the untreated radioactive granules primary GBM(P<0.01).Conclusion The resistance of recurrent GBM to radiotherapy may be associated with the overexpression of RAD18 protein.

Objective:Whether extralevator abdominoperineal excision (ELAPE) improves survival and safety remains controversial.Systematic review of all comparative studies to define the superiority of ELAPE to conventional abdominoperineal excision (APE).Methods:Corresponding data,with case-control studies or cohorts regarding intraoperative perforation rate,the local recurrence rate and postoperative complications in the ELAPE group and the APE group,were retrieved from PubMed,Embase,the Cochrane Library,Chinese Biomedical Literature (CMB),VIP,China National Knowledge Infrastructure (CNKI),and Wanfang Database.Meta-analysis was performed by using RenMan 5.2.Results:A total of 10 articles were included.Intraperative perforation rate (MD=0.54,95％ CI 0.31 to 1.39,P=0.03),local recurrence rate (MD=0.30,95％ CI 0.21 to 0.42,P＜0.001) in the ELAPE group was significantly lower than that in the APE group.The difference in positive margin rate between the 2 groups was not statistically significant (P=0.07).Conclusion:Through gap repair of episiotomy and individualized therapy can improve ELAPE postoperative quality of life.ELAPE shows certain advantages in treating lower rectal cancer comparing to APE,but it should pay attention to individualized treatment.More studies through large sample multi-center,medium and long term randomized design are necessary to determine the effect of surgery on tumor.

Objective To investigate the expression of excision repair cross-complementation group 1 (ERCC1) and ribonucleotide reductase subunit M1 (RRM1) in patients with esophageal squamous cell carcinoma,and the relationship between the expression of ERCC1 and RRM1 genes and chemotherapy sensitivity.Methods A total of 77 patients with esophageal squamous cell carcinoma admitted in hospital from Jan.2008 to Dec.2012 were recruited.ERCC1 and RRM1 mRNA levels in the cancerous tissue were detected by real-time fluorescent quantitative RT PCR.The relationship between short-term effects of chemotherapy and ERCC1 and RRM1 mRNA levels was analyzed.Results Levels of mRNA in stages Ⅰ a,Ⅰ b,Ⅱ a,and Ⅱ b were (0.578±0.081),(0.560±0.084),(0.521±0.080),(0.464±0.091) for ERCC1 and(0.511±0.089),(0.539± 0.086),(0.584±0.092),(0.637±0.101)for RRM1,respectively.As the clinical stage advanced,the ERCC1 mRNA level declined and the RRM1 mRNA level increased (t=2.679 and 2.952,P =0.034 and 0.025,respectively).Levels of mRNA in patients with complete remission,partial remission,stable disease and progressive disease were (0.487 ± 0.097,0.511 ± 0.095,0.552 ± 0.086,0.568 ± 0.088) for ERCC1 and(0.504±0.091,0.544±0.095,0.595±0.093,0.616±0.097) for RRM1,respectively.The clinical effect of chemotherapy was negatively correlated with mRNA expression of ERCC1 and RRM1 (r=0.567,P=0.032).Conclusions Levels of ERCC1 and RRM1 mRNA expression are correlated with the staging of esophageal squamous cell carcinoma and chemotherapy sensitivity,and can be used as a predictive parameter for chemotherapy sensitivity in patients with esophageal squamous cell carcinoma.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Diseases ; diagnosis ; Bile Duct Neoplasms ; diagnosis ; Cholangiopancreatography, Endoscopic Retrograde ; Cytodiagnosis ; Cytological Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Diseases ; diagnosis ; Pancreatic Neoplasms ; diagnosis

In this study, the rescue effect of receptor activator for nuclear factor-kappaB ligand (RANKL) on zoledronate acid (ZOL) induced inhibition of osteoclastogenesis and gene expression of NF-kappaB p50 and c-Jun was investigated. Mice calvarial osteoblasts (OBs) were harvested and co-cultured with RAW264.7 cells and the cells were divided into 4 groups and received treatment with ZOL and RANKL, either single or combined. The formation of multi-nucleated osteoclast (OC) was examined and gene expression of NF-kappaB p50 and c-Jun was detected. Group B (ZOL) showed least multi-nucleated OC and resorption lacunae among the 4 groups (P < 0.05 or P < 0.01) and it was followed by group C (ZOL+RANKL). Group D (RANKL) showed highest OC and resorption lacunae while it was similar to Group A (control) (P > 0.05). Gene expression of NF-kappaB p50 and c-Jun was the lowest in group B (P < 0.05 or P < 0.01) among the four groups and was significantly increased in group C when compared with group B (P < 0.05). Group A and D showed highest gene expression and they were similar to each other (P > 0.05). This study suggest that RANKL might partly rescue ZOL induced inhibition of osteoclastogenesis, and the effect of RANKL and ZOL on osteoclastogenesis may be mediated by NF-kappaB p50 and c-Jun.
Animals ; Bone Resorption ; drug therapy ; Cell Line ; Diphosphonates ; pharmacology ; Gene Expression ; Imidazoles ; pharmacology ; Mice ; NF-kappa B p50 Subunit ; metabolism ; Osteoblasts ; drug effects ; Osteoclasts ; drug effects ; Proto-Oncogene Proteins c-jun ; metabolism ; RANK Ligand ; pharmacology

Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

Objective: To determine total phenylethanoid glycoside and acteoside in Plantago Herba to provide reference for evaluating the quality of medicinal materials.Methods: With acteoside as the control sample, a UV visible spectrophotometric method was used to determine total phenylethanoid glycosides in Plantago Herba.An HPLC method was applied to determine acteoside in Plantago Herba , and the conditions were as follows: an ODS2 C 18 (150 mm× 4.6 mm ,5 μm) chromatographic column was used with acetonitrile-0.1% formic acid (13∶87) as the mobile phase at a flow rate of 1.0 ml·min-1 , the detection wavelength was 332nm, the column temperature was 30℃, and the sample volume was 10 μl.Results: The reference solution and the sample solution had the maximum absorption at 332 nm, and the linear relationship was good within the range of 0.003 1-0.155 0 mg·ml-1 (r=0.999 5).The content of total benzene alcohol glycosides in 3 batches of samples was 2.73% , 2.61% and 2.84% , respectively;acteoside over the range of 0.000 6-0.155 0 mg·ml-1 (r=0.999 1) showed a good linear relationship with peak area,the sample recovery was 98.5% and the RSD was 1.6% (n =6), and the acteoside content in 3 batches of samples respectively was 0.54% , 0.51% and 0.56%.Conclusion: The method is simple, accurate and reproducible, and can be used for the determination of total phenylethanoid glycosides and acteoside in Plantago Herba.

Abstract: Objective　To investigate the prevalence and genetic characterization of Blastocystis in primary and middle school students in Baisha Li Autonomous County, Hainan Province, in order to understand the infection status of Blastocystis and its subtype distribution characteristics in this area. Methods　From March to November 2021, fecal samples were collected from two primary and middle schools in Baisha Li Autonomous County. Nested PCR targeting the SSU rDNA was employed in this study, sequence analysis were performed to determine the prevalence and subtype. A neighbor-joining tree was built using Mega 7. Meanwhile, the risk factors of the Blastocystis infection among different grades and genders were evaluated. Results　The infection rate of Blastocystis was 4.1% (13/314), there was no statistical difference in infection rates among genders and grades (P>0.05). Sequence analysis revealed that three Blastocystis subtypes were identified, namely ST3 (n=7), ST7 (n=4) and ST1 (n=2), all of which have zoonotic potential. Conclusions　This is the first report of the identification of Blastocystis in humans in Hainan at the subtype level, and provide the basic data for the prevention and control of Blastocystis in this area. The zoonotic subtypes identified in this area indicated more studies should be taken in humans and various animals, to better evaluate the transmission of Blastocystis and provide scientific support for the prevention and control of Blastocystis.

Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method. Use the Sevage method to remove the protein in the crude polysaccharide. The structural characteristics of the polysaccharide were determined by FT-IR spectra.The RM-1 and HeLa cells were divided into control group and different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )of ginseng polysaccharide groups. The survival rates of the cells in various groups were detected by MTT method. The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )on the cancer cells were determined by CTL test. Results The extraction rate of ginseng polysaccharide was 8.7 6% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between different concentrations of ginseng polysaccharide groups after treated for 24 h compared with control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly (P<0.05)compared with control group;with the increase of ginseng polysaccharide concentration, the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg·L-1 ,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.

Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.