Expression and significance of NGX6 gene in human hepatocellular carcinoma / 中南大学学报(医学版)

OBJECTIVE@#To examine the expression of NGX6 gene and to investigate its association with the clinico-pathological characteristics in hepatocellular carcinoma(HCC).@*METHODS@#Samples from 45 patients were divided into the hepatocellular carcinoma tissue group and the matched paracancerous tissue group. Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of NGX6 gene in the hepatocellular carcinoma and the surrounding normal tissue specimens.@*RESULTS@#The positive rates of NGX6 in the hepatocellular carcinoma and the matched paracancerous tissues were 35.5%(16/45)and 77.8%(35/45), and the ratios of NGX6/G3PDH mRNA were 0.245+/-0.060 and 0.352+/-0.113.There was significant difference in the 2 groups (P<0.05). The expression of NGX6 gene was related to TNM staging (chi2=6.106,P=0.042)and lymph node metastasis(chi2=5.237,P=0.022)in hepatocellular carcinoma.@*CONCLUSION@#There is positive expression of NGX6 in the hepatocellular carcinoma and the matched paracancerous tissues. The low-expression or non-expression of NGX6 gene plays an important role in the gene transcription level in hepatocellular carcinoma.The expression of NGX6 gene is related with TNM staging and lymph node metastasis in hepatocellular carcinoma. Expression of NGX6 might be used as an early indicator of the occurrence and development of hepatocellular carcinoma.

Efficacy of 4 kinds of triple strategy for Helicobacter pylori eradication / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the efficacy and safety of 4 kinds of triple strategy of Helicobacter pylori (Hp) eradication.@*METHODS@#A total of 307 patients who suffered from Hp infection, confirmed by rapid urease test (RUT) and 14C-urea breath test (UBT),were randomly divided into 4 groups. Each group had 80, 76, 77, and 74 patients respectively. Group A was treated with rabeprazole, clarithromycin, and furazolidone (RCF); Group B with ranitidine bismuth citrate, clarithromycin, and furazolidone (BCF); Group C with rabeprazole, amoxicillin, and furazolidone (RAF); while Group D with ranitidine bismuth citrate, amoxicillin, and furazolidone (BAF). Hp was detected by RUT and UBT at 4 weeks after later treatment.@*RESULTS@#Hp eradication rates of group A,B,C, and D were 90.0%,67.1%,62.3%,and 45.9%,respectively. The difference between Group A and Group B, Group A and Group C was significant (P<0.05). Eradication rate of Group B and C was higher than that of Group D (P<0.05). There was no statistical difference between the eradication rate of Group B and C, and among the side effects of the 4 groups.@*CONCLUSION@#The strategy of RCF was the best among the 4 triple strategy of Hp eradication, which can be used clinically.

Helicobacter pylori infection and changes of cell gap junction of gastric epithelial cells in patients with gastric cancer and precancerous lesion / 中南大学学报(医学版)

OBJECTIVE@#To observe the changes of cell gap junction ultrastructure of gastric epithelial cells in patients with gastric cancer(GC) and precancerous lesion(PL),and to investigate the relation between these changes and H.pylori infection.@*METHODS@#Seventy patients with GC, 88 with PL, and 33 with chronic superfial gastritis (CSG) were studied. H.pylori was detected by rapid urease test,basic fuchsin stain and 14C-urea breath test. The CagA gene of H.pylori was determined by polymerase chain reaction(PCR).The cell gap junction ultrastructure was observed under transmission electronic microscope.@*RESULTS@#Length of junction/unit perimeter of gastric epithelial cells in patients with PL was smaller than that in CSG patients, and the smallest width of the intercellular space was bigger than that in CSG patients. The number of cell junction, the number of junction/unit perimeter, and the length of junction/unit perimeter in patients with GC were all smaller than those in patients with CSG or PL, and its smallest width of the intercellular space was bigger than that in patients with CSG. In patients with GC, the number of cell junction, the number of junction/unit perimeter and the length of junction/unit perimeter in CagA+ H.pylori group were smaller than those in CagA(-) H.pylori group, and its smallest width of the intercellular space was bigger than that in CagA(-) H.pylori group. In PL patients, the intercellular space decreased, and the length of cell junction of gastric epithelial cells became bigger after H.pylori eradication. The length of junction/unit perimeter in patients of H.pylori eradication was bigger than that in patients without eradication, and the smallest width of the intercellular space was smaller than that in patients without eradication.@*CONCLUSION@#The changes of cell gap junction of gastric epithelial cells in patients with GC and PL are associated with H.pylori infection especially CagA+ H.pylori infection. Eradication of H.pylori can promote the formation of cell junction.

Expression and SNP analysis of BRD7 gene in acute leukemia cells / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the expression level of BRD7 gene in bone marrow mononuclear cells (BMNCs) in patients with acute leukemia (AL) and to analyze BRD7 single nucleotide polymorphism(SNP).@*METHODS@#RT-PCR was used to detect BRD7 expression in patients with AL and normal bone marrow subjects. Single-strand conformation polymorphism and DNA sequence analysis were also used to identify BRD7 mutation or SNP to investigate the relation between BRD7 and AL.@*RESULTS@#BRD7 mRNA in BMNCs from 52 patients with AL and 30 control subjects was expressed. The mRNA relative expression of BRD7 in patients with AL was higher than that of the control group (P=0.001). Three SNPs (C657A,C495T and A737G) in BRD7 gene coding region (447 approximately 844 bp) were found, and A737G was coupled with C495T . The allele frequencies of SNP C657A were not significantly different between AL and the control group. The genotype and the allele frequencies of the 2 coupled SNPs were significantly different (P0.05).@*CONCLUSION@#Expression of BRD7 gene is up-regulated in AL cells. The 2 coupled SNPs (C495T and A737G ) in BRD7 gene coding region (447 approximately 844 bp) are correlated with AL, indicating that SNPs may be one of the genetic susceptibility factors of AL.

Effects of NGX6 on the transcriptional activation of beta-catenin/TCF/LEF in Wnt/beta-catenin signal pathway / 中南大学学报(医学版)

OBJECTIVE@#To explore the effects of NGX6 on the transcriptional activation of beta-catenin/TCF/LEF in Wnt/beta-catenin signal pathway, and to identify the role of NGX6 in Wnt signal pathway.@*METHODS@#The eukaryotic expression vector pcDNA3.1(+)-beta-catenin (WT) was constructed. pcDNA3.1(+)-beta-catenin (WT) and pCMV-myc-NGX6 were cotransfected to COS-7 and the transcriptional activity of TCF/LEF was detected by TCF-4 luciferase report system. Without extro-genous beta-catenin, pCMV-myc-NGX6 was transfected alone to COS-7 and colon cancer cell line SW620, and the transcriptional activity of TCF/LEF was detected by TCF-4 luciferase report system, and then the expression of nucleus beta-catenin and TCF-4 was detected by Western blot.@*RESULTS@#The eukaryotic expression vector pcDNA3.1(+)-beta-catenin (WT) was successfully constructed. The activation of TCF-4 luciferase report gene in the cotransfection group in COS-7 was less than that in NGX6 alone transfection group (P<0.05). The activation of TCF-4 luciferase report gene in NGX6 alone transfection group without extro-genous beta-catenin was less than that in pCMV-myc transfection group in COS-7 and SW620. The expression of beta-catenin and TCF-4 was decreased after the NGX6 transfection in COS-7 and SW620 cells.@*CONCLUSION@#NGX6 can inhibit the transcriptional activation of beta-catenin/TCF/LEF in Wnt signal pathway by its negative regulation in the nuclear translocation of beta-catenin.

Effect of CagA(+) helicobacter pylori strain on the expression of connexin 43 and cell proliferation in BGC-823 cells / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of CagA(+) Helicobacter pylori(H.pylori)strain and anti-H.pylori drugs on the expression of connexin 43(Cx43) and cell proliferation of BGC-823 cells in vitro,and to investigate the relation between the changes of Cx43 expression, cell proliferation of BGC-823 cells and CagA(+)H.pylori.@*METHODS@#BGC-823 cells were co-cultured with CagA(+) H.pylori strain(NCTC J99) or CagA(-) H.pylori strain(NCTC 12908)at bacteria/cells ratio of 20:1,100:1 and 500:1 for 24 hours and 48 hours respectively. anti-H.pylori drugs was given in the group co-cultured at bacteria/cells ratio of 100:1 after 16 hours. In the control group, BGC-823 cells were cultured for 24 hours and 48 hours respectively,but without H.pylori or antij H.pylori drugs. Immunocytochemical SABC method and the image analysis of the computer were applied to detect the changes of Cx43 expression in BGC-823 cells. The cell proliferation was examined by methyl tetrazolium (MTT) method.@*RESULTS@#(1)The expression of Cx43 in the control group after cultivation for 48 hours was higher than that for 24 hours (P0.05). Cx43 expression in the groups co-cultured with CagA(-) H.pylori strain at the ratio of 100:1 and 500:1 was lower than that in the control group, and Cx43 expression at the ratio of 500:1 was lower than that at the ratio of 20:1 for 24 hours and 48 hours. Cx43 expression increased after the intervention with anti-H.pylori drugs for 48 hours. (2) In the groups co-cultured with CagA(+)H.pylori strain, the optical density value of MTT indicated that the cell proliferation at the bacteria/cells ratio of 100:1 was higher than that in the control group, but no significant difference was found in other two groups co-cultured for 24 hours. After co-culturing for 48 hours, the cell proliferation at the bacteria/cells ratio of 20:1 and 100:1 was significantly accelerated, while the cell proliferation at 500:1 was inhibited. In the groups co-cultured with CagA(-) H.pylori strain,there was no change in the cell proliferation. Intervention with anti-H.pylori drugs could suppress the cell proliferation.@*CONCLUSION@#CagA(+) H.pylori can down-regulate the expression of Cx43 in BGC-823 cells,which is related to the reaction time and the density of H.pylori. Low density of CagA(+)H.pylori suspensions can accelerate the proliferation of BGC-823 cells, while high density can suppress the cell proliferation. The CagA(-) H.pylori has no effect on the cell proliferation. Intervention with anti-H.pylori drugs can up-regulate the expression of Cx43,and suppress the cell proliferation of BGC-823 cells.

Effect of various uses of propofol on the upper gastrointestinal endoscopy / 中南大学学报(医学版)

OBJECTIVE@#To investigate the efficacy and security of different uses of propofol on the sedation during the upper gastrointestinal endoscopic procedures.@*METHODS@#Four hundred patients who underwent gastroscopy received midazolam and propofol as sedation. Patients were divided to 4 groups with different intervals between midazolam and propofol: Group A and D with the interval of 30 seconds to 1 minute, Group B and C with 3 to 5 minute interval. All patients were premedicated with midazolam and propofol at 16 approximately 25 mg/10s (Group A and B) and 6 approximately 7 mg/10s (Group C and D).@*RESULTS@#The doses of propofol of Group A,B,C, and D were (111.90+/-22.43),(102.20+/-15.99),(73.05+/-13.08) and (80.90+/-17.36)mg respectively, with significant difference(P<0.01). The time of return to consciousness decreased markedly in Group C and D [(9+/-1), (10+/-2)min ], and that of Group A and B was [(14+/-5), (13+/-3)min ]. There was significant difference between Group C, D and Group A, B(P<0.01).@*CONCLUSION@#The dose of propofol and the time of return to consciousness depend on the rate of administration and the interval between midazolam and propofol. Appropriate rate and interval can produce safer and more effective sedation for the upper gastrointestinal endoscopic procedure.

Antibiotic resistance of Helicobacter pylori / 中南大学学报(医学版)

OBJECTIVE@#To examine the infection and bacteria resistance of Helicobacter pylori (H.pylori) to clarithromycin and furazolidone,to determine whether the antibiotic resistance is primary or secondary, and to decide if a new H.pylori infection plays a role in eradication failures.@*METHODS@#Twenty one H.pylori had been isolated from human biopsy specimens, and antimicrobial susceptibility testing was performed. DNA fingerprints were generated using random amplification polymorphic DNA (RAPD) to determine the identity of H.pylori before and after the eradication therapy.@*RESULTS@#Eight bacteria resisted against clarithromycin, and one against furazolidone, with the resistant rates 38.1% and 4.8% respectively. The number of primary antibiotic resistance, secondary resistance and new infection was 1 for each.@*CONCLUSION@#Resistance to clarithromycin is more common compared with that to furazolidone. Development of primary and secondary resistance to clarithromycin occurs as a rule in eradication failures. New H.pylori infection plays a role in eradication failures.

Intrasplenic tumor model of nude mice in the anti- metastasis roles of NGX6 gene against colon cancer / 中南大学学报(医学版)

OBJECTIVE@#To establish a liver metastasis model of nude mice in colon cancer so as to determine the function of NGX6.@*METHODS@#The cells of Group HT-29, pcDNA3.1(+)/HT-29, and pcDNA3.1(+)/NGX6/HT-29 were implanted into the spleen of nude mice, respectively. Everyday we measured the weight of the nude mice and observed their ingestion, movement and mental status. The nude mice were killed after 45 days, and the effect of NGX6 on the malignant behavior of HT-29 was assessed by this experiment.@*RESULTS@#In contrast to the other two groups, the metastasis in the liver and xenograft tumor in the spleen of pcDNA3.1(+)/NGX6/HT-29 group was significantly reduced (P<0.01).@*CONCLUSION@#The metastasis of HT-29 colon cancer cell line was significantly inhibited by NGX6 gene. This model of liver metastasis in the nude mice is a proper model to determine the anti-metastasis mechanism of NGX6 gene.

Contrast enhanced power Doppler in evaluating the angiogenesis in hepatocellular carcinoma / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the role of contrast enhanced power Doppler in evaluating tumor angiogenetic activity.@*METHODS@#Thirty-two patients with hepatocellular carcinoma were analysed. Flow signals of hepatocellular carcinoma were observed by power Doppler imaging after the injection of contrast agent, and then the relative perfusion rate and blood flow were assessed. The microvessel density (MVD) and vascular endothelial growth factor(VEGF) were assessed by immunohistochemical method. The relationship between the relative perfusion rate,blood flow, MVD,VEGF was studied.@*RESULTS@#The relative perfusion rate in the tissues with positive expression of VEGF was significantly higher than that in the tissues with negative expression of VEGF in hepatocellular carcinoma. There was correlation between the relative perfusion rate, blood flow grade and MVD(P<0.05). The expression of VEGF was positively related to the relative perfusion rate and blood flow grade(P<0.05).@*CONCLUSION@#Contrast enhanced power Doppler is useful in evaluating the tumor angiogenetic activity.

Effect of prostaglandin E1 on the expression of tissue inhibitor of metalloproteinase-1 in experimental liver fibrosis rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of prostaglandin E1 (PGE1) on the expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) in experimental liver fibrosis rats.@*METHODS@#The liver fibrosis model was established by carbon tetrachloride. Rats were divided into a control group and PGE1-treated group. The pathological changes of the liver tissue from the two groups, the semi-quantitative analysis of hepatitic activity in HE stain sections, the pathological image quantitative analysis of the fibrosis degree, TIMP-1 positive cells, and the content of collagen were synthetically analysed.@*RESULTS@#The mark changes of liver pathology in HE stain sections were that the degree of hepatitic activity in the PGE1-treated group was obviously lower than that in the control group (P < 0.05). The fibrosis degree, TIMP-1 positive cells and the collagenous fibers decreased in the PGE1-treated group (P <0.05).@*CONCLUSION@#PGE1 has an anti-hepatofibrosis effect in the experimental rats, the inflammation of liver is light, and the proliferation of collagenous fibers can be restrained, whose mechanism is probably associated with the suppression of TIMP-1 expression caused by PGE1.

Two-stage administration sedation method for gastroscopy in hypoxia predisposed patients / 中南大学学报(医学版)

OBJECTIVE@#To explore the safety and the efficacy of two-stage administration sedation method (with propofol plus midazolam) in the analgesia gastroscopy for hypoxia predisposed patients.@*METHODS@#Sixty-four hypoxia predisposed patients (with snore or expectoration) were divided into two groups and received sedation respectively with two-stage administration method and routine continuum administration method before gastroscopy. We analyzed the alteration of SBP, DBP, SpO2, sedation level, the degree of pharyngeal malaise, recovery time, dose of propofol,the lethe degree of upset, and procedure of gastroscopy to evaluate the safety and the efficacy of the two methods in the analgesia gastroscopy for hypoxia predisposed patients.@*RESULTS@#Compared with routine continuum administration sedation method, two-stage administration sedation method had little influence on SpO2, so it reduced the accident of respiratory arrest (P 0.05), and the lethe degree of gastroscopy procedure in two-stage administration method group was significantly lower than that of routine continuum adminis- tration method group (P <0.01).@*CONCLUSION@#The two-stage administration sedation method is a new effective and safe sedation method for gastroscopy in the hypoxia predisposed patients.

Studies on the relationship between D6S1581, a high frequency allele imbalance locus, and genetic susceptibility to nasopharyngeal carcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581.</p><p><b>METHODS</b>Genescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed.</p><p><b>RESULTS</b>D6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005).</p><p><b>CONCLUSION</b>These results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.</p>

Genetic polymorphism of 8 STR loci on short arm of chromosome 3 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area.</p><p><b>METHODS</b>Blood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer.</p><p><b>RESULTS</b>Ninety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black.</p><p><b>CONCLUSION</b>These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application.</p>

Polymorphism of two novel SNPs, which locate on chromosome 9p21-22, in Han Chinese of Hunan / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To search novel SNPs in exons and regulatory regions of CDKN2A and two novel putative tumor suppressor genes NGX6 and UBAP1, which all reside on chromosome 9p21-22.</p><p><b>METHODS</b>The exons and regulatory regions of those genes were amplified and sequenced in 96 subjects.</p><p><b>RESULTS</b>Two novel SNPs were found, one resides on the sixth exon of UBAP1 gene and the other on the fourth exon of CDKN2A gene. Two novel SNPs were submitted to the dbSNP database, and their access ID are rs3135929 and rs3088440. The polymorphic information contents of them are 0.102 and 0.213 respectively. There is linkage equilibrium between them, and the polymorphic information content of their haplotype is 0.302, higher than any of them individually.</p><p><b>CONCLUSION</b>The polymorphic information content can be improved by using haplotype analysis of several SNPs.</p>