Flavonoid-Rich Extract of Oldenlandia diffusa (Willd.) Roxb. Inhibits Gastric Cancer by Activation of Caspase-Dependent Mitochondrial Apoptosis / 中国结合医学杂志

OBJECTIVE@#To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells, determine the action mechanisms in association with the mitochondrial dependent signal transduction pathway that controls production of reactive oxygen species (ROS), and evaluate the pharmacodynamics of a mouse xenotransplantation model to provide a reference for the use of flavonoids in prevention and treatment of gastric cancer.@*METHODS@#Flavonoids were extracted by an enzymatic-ultrasonic assisted method and purified with D-101 resin. Bioactive components were characterized by high-performance liquid chromatography. Cell lines MKN-45, AGS, and GES-1 were treated with different concentrations of flavonoids (64, 96, 128, 160 µg/mL). The effect of flavonoids on cell viability was evaluated by MTT method, and cell nuclear morphology was observed by Hoechst staining. The apoptosis rate and cell cycle phases were measured by flow cytometry, the production of ROS was detected by laser confocal microscope, the mitochondrial membrane potential (MMP) were observed by fluorescence microscope, and the expression of apoptotic proteins related to activation of mitochondrial pathway were measured by immunoblotting. MKN-45 cells were transplanted into BALB/c nude mice to establish a xenograft tumor model. Hematoxylin and eosin staining was used to reveal the subcutaneous tumor tissue. The tumor volume and tumor weight were measured, the expression levels of proliferation markers proliferating cell nuclear antigen (PCNA) and Ki-67 were detected by immunohistochemistry, and the expression levels of CA72-4 were measured by enzyme linked immunosorbent assay.@*RESULTS@#Oldenlandia diffusa flavonoids inhibited proliferation of MKN-45 and AGS human gastric cancer cells, arrested the cell cycle in G1/S phase, induced accumulation of ROS in the process of apoptosis, and altered MMP. In addition, flavonoids increased Apaf-1, Cleaved-Caspase-3, and Bax, and decreased Cyclin A, Cdk2, Bcl-2, Pro-Caspase-9, and Mitochondrial Cytochrome C (P<0.05). The MKN-45 cell mouse xenotransplantation model further clarified the growth inhibitory effect of flavonoids towards tumors. The expression levels of PCNA and Ki-67 decreased in each flavonoid dose group, the expression level of CA72-4 decreased (P<0.05).@*CONCLUSION@#Flavonoids derived from Oldenlandia diffusa can inhibit proliferation and induce apoptosis of human gastric cancer cells by activating the mitochondrial controlled signal transduction pathway.

Overexpression of CLEC5A inhibits cell proliferation and metastasis and reverses epithelial-mesenchymal transition in hepatocellular carcinoma / 南方医科大学学报

OBJECTIVE@#To evaluate the effects of CLEC5A expression level on cell proliferation, migration and invasion and epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma (HCC) and explore the role of CLEC5A in the tumorigenesis and progression of HCC.@*METHODS@#The expression level of CLEC5A was detected in 50 pairs of HCC and adjacent tissues using immunohistochemical staining, and its association with clinicopathological parameters of HCC patients was analyzed. Cultured HCC cell line SK-HEP-1 was transfected with a lentiviral vector overexpressing CLEC5A, and the transfection efficiency was verified using real-time fluorescence quantitative PCR and Western blotting. The changes in proliferation, migration and invasion abilities of the transfected cells were analyzed using CCK-8, 5-ethynyl-29-deoxyuridine (EdU) and Transwell assays, and EMT of the cells was determined using Western blotting.@*RESULTS@#The protein expression level of CLEC5A was significantly lower in HCC tissues than in the adjacent tissues (P < 0.001). The expression level of CLEC5A was significantly correlated with tumor size (P=0.008), tumor number (P=0.010), histological differentiation (P=0.016), microvascular invasion (P=0.024) and BCLC stage (P=0.040). In SK-HEP-1 cells, overexpression of CLEC5A obviously inhibited the cell proliferation, migration and invasion and reversed EMT phenotype of the cells.@*CONCLUSION@#CLEC5A is a potential HCC suppressor gene and may serve as a promising therapeutic target for HCC.

Cloning and sequence analysis of the DHBV genome of the brown ducks in Guilin region and establishment of the quantitative method for detecting DHBV / 病毒学报

Brown ducks carrying DHBV were widely used as hepatitis B animal model in the research of the activity and toxicity of anti-HBV dugs. Studies showed that the ratio of DHBV carriers in the brown ducks in Guilin region was relatively high. Nevertheless, the characters of the DHBV genome of Guilin brown duck remain unknown. Here we report the cloning of the genome of Guilin brown duck DHBV and the sequence analysis of the genome. The full length of the DHBV genome of Guilin brown duck was 3 027bp. Analysis using ORF finder found that there was an ORF for an unknown peptide other than S-ORF, PORF and C-ORF in the genome of the DHBV. Vector NTI 8. 0 analysis revealed that the unknown peptide contained a motif which binded to HLA * 0201. Aligning with the DHBV sequences from different countries and regions indicated that there were no obvious differences of regional distribution among the sequences. A fluorescence quantitative PCR for detecting DHBV was establishment based on the recombinant plasmid pGEM-DHBV-S constructed. This study laid the groundwork for using Guilin brown duck as a hepatitis B animal model.

Effect of percutaneous transhepatic cholangial drainage with bile reinfusion and enteral nutrition via the nasojejunal tube on visceral protein and hepatic function / 南方医科大学学报

<p><b>OBJECTIVE</b>To discuss the clinical value of percutaneous transhepatic cholangial drainage (PTCD) combined with nasojejunal tube for bile reinfusion and enteral nutrition for patients with malignant obstructive jaundice.</p><p><b>METHODS</b>Forty patients with malignant obstructive jaundice were randomly divided into bile reinfusion group (n=20) and exclusive external drainage group (control group, n=20), and the clinical data concerning the hepatic function, visceral protein and postoperative complications of the patients were collected.</p><p><b>RESULTS</b>In both of the two groups, the levels of ALT, AST, and TB-2 reduced significantly after the operation as compared with the preperative levels (P<0.05), and no significant difference was found in the postoperative hepatic function between the two groups (P>0.05). The postoperative levels of the visceral proteins such as ALB, TRF and PRE increased significantly after the operation (P<0.05), and the changes in ALB and PRE were comparable between the two groups (P>0.05). TRF was significantly higher in bile reinfusion group than in the control group.</p><p><b>CONCLUSION</b>PTCD combined with bile reinfusion and early enteral nutrition via the nasojejunal tube may facilitate the recovery of hepatic function and visceral proteins in patients with malignant obstructive jaundice.</p>

Study on SNP polymorphism of mitochondrial DNA D-loop region from Nu ethnic population in Yunnan of China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the mitochondrial single nucleotide polymorphism (SNP)of Chinese Nu ethnic population from Yunnan region of China and to provide basic database for ethnic origin investigation and forensic purpose.</p><p><b>METHODS</b>Genomic DNA from the whole blood of 87 unrelated individuals was extracted by standard chelex-100. The sequence polymorphism was analyzed by PCR-based assay and using ABI 3730 Analyzer to detect many number of relatively common point mutations.</p><p><b>RESULTS</b>Sixty-two SNP loci were observed among them with 492 point mutations and 59 haploids identified in mitochondrial DNA hypervariable region I (mtDNA HVSI). The gene diversity was estimated to be 0.9675,and the random match probability was calculated to be 0.0437.</p><p><b>CONCLUSION</b>The result suggests that mtDNA HVSISNP database of Nu ethnic population can be a useful tool for forensic identity and original research.</p>

Sequence-length polymorphism research of C-stretch in mitochondrial DNA hyper-variant region / 法医学杂志

OBJECTIVE@#To investigate the polymorphism of C-stretch in mitochondrial DNA hyper-variant region and the values in forensic science.@*METHODS@#The gene frequency were detected in such two loci of nt16180 and nt310 by using PCR-sequencing technique with two pair of primers according to the reference.@*RESULTS@#The length of PCR products were 807 bp and 962 bp respectively. Seven genotypes in each locus were detected. AAAACCCCCTCCCC allele amounted to 87.72% of all nt16180 genotype, while AAAACCCCCCCCCCCCC was firstly reported in Han population. CCCCCCCCTCCCCCC allele amounted to 60.53% of all nt310 genotype. 15 haplotypes were detected among 114 samples. The GD value was 0.6309. The AAAACCCCCTCCCC-CCCCCCCCTCCCCCC haplotype was the most high frequency and amounted to 57.89%.@*CONCLUSION@#The obtained basic genetic data in two loci of nt16180 and nt310 of mtDNA will be useful for forensic science.

Allele distribution of five X-chromosome short tandem repeat loci in Ewenke population of north China / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To study the allele genetic polymorphism of five short tandem repeat (STR) loci on X-chromosome in Ewenke population of north China and to provide basic data for forensic identification.</p><p><b>METHODS</b>Genomic DNA was extracted from EDTA-whole blood of Ewenke population by Chelex-100. The DNA samples were amplified by PCR and were analyzed by polyacrylamide gel electrophoresis and silver staining. The sequence length variations of DXS6799, DXS8378, DXS101, HPRTB, and DXS6789 loci on X-chromosome in 98 unrelated Ewenke individuals were investigated.</p><p><b>RESULTS</b>All five loci analyzed showed high polymorphism and genetic stability. The data of the five X-chromosome STR loci in Ewenke ethnic group of China was in accordance with Hardy-Weinberg equilibrium by Chi-square test.</p><p><b>CONCLUSION</b>Allele polymorphism of five X-chromosome STR loci can be used as a genetic marker for forensic identification and population genetic research.</p>