The effect of Glut1 and c-myc on prognosis in esophageal squamous cell carcinoma of Kazakh and Han patients.

AIM: Glucose transporter type 1 (Glut1) plays a crucial role in cancer-specific metabolism. We explored the expression of Glut1 and c-myc, the relationship between them and the effect of Glut1, c-myc on prognosis in esophageal squamous cell carcinoma. MATERIALS & METHODS: Immunohistochemistry was used to examine the expression of Glut1 and c-myc. χ2 test analyzes the relationship between c-myc, Glut1 and pathological parameters. Spearman correlation analyzes the relationship between c-myc and Glut1. Survival analysis was used to investigate the effect of Glut1 and c-myc on prognosis. RESULTS: Glut1 positivity was associated with tumor size (p < 0.01), depth of invasion (p = 0.021), Tumor, Node, Metastasis stage (IA+IB,II+IIB,IIIA+IIIB,IVA+IVB; p = 0.004), lymph node metastasis (p = 0.002) and nerve invasion (p = 0.050). C-myc positivity was associated with tumor location (p = 0.015), depth of invasion (p = 0.022) and lymph node metastasis (p = 0.035). There was a positive correlation between c-myc and Glut1 (r = 0.321). Patients with Glut1 c-myc co-expression had poorer prognosis. CONCLUSION: Inhibiting Glut1 c-myc co-expression may improve the prognosis of esophageal squamous cell carcinoma.

Genetic variations in the KCNJ5 gene in primary aldosteronism patients from Xinjiang, China.

BACKGROUND: Primary aldosteronism (PA) is the most common endocrine form of secondary hypertension, and one of the most common subtypes of sporadic PA is aldosterone-producing adenoma (APA). Recently, two somatic mutations of the KCNJ5 gene were implicated in APA, and two germline mutations were associated with familial hyperaldosteronism III. OBJECTIVES: This case-control study was designed to investigate the relationship between genetic variations in the KCNJ5 gene and sporadic PA patients in Xinjiang, China. METHODS: Five common single nucleotide polymorphisms (SNPs) of the KCNJ5 gene (rs6590357, rs4937391, rs3740835, rs2604204, and rs11221497) were detected in patients with sporadic PA (n = 235) and essential hypertension (EH; n=913) by the TaqMan polymerase chain reaction method. RESULTS: The EH group and the PA group showed significant differences in the distributions of genotypes and alleles of rs4937391 and rs2604204 in total and male subjects (P<0.05), as well as rs3740835 in male subjects (P<0.05). However, only the association between the rs2604204 genotype and male sporadic PA remained significant after Bonferroni's correction (P<0.01). Furthermore, logistic regression analysis demonstrated that the CC genotype of rs2604204 was a risk factor for male patients with sporadic PA, after adjusting for age and body mass index (odds ratio=2.228, 95% CI: 1.300-3.819, P=0.004). CONCLUSION: The genetic variant rs2604204 of KCNJ5 is associated with sporadic PA in Chinese males, suggesting that KCNJ5 may be involved in the pathogenesis of sporadic PA in these particular patients.

[Etiology analysis for hospitalized hypertensive patients: 10 years report from the department of hypertension (1999 - 2008)].

OBJECTIVE: To analyze etiology of hospitalized hypertensive patients in the department of hypertension from 1999 to 2008. METHODS: This retrospective study was performed to analyze the etiology of hospitalized hypertensive patients in department of hypertension and to show the distribution change of hypertension from 1999 to 2008. RESULTS: (1) There were 5867 (75.1%) patients with essential hypertension and 1942 (24.9%) patients with secondary hypertension (SH). (2) The prevalence rate of SH increased significantly during the 10 years (χ(2) = 387.621, P < 0.001) and was higher in 2008 than in 1999 (39.3% vs. 9.5%, P < 0.05). The prevalence of obstructive sleep apnea syndrome (OSAS) and primary aldosteronism (PA) in 2008 increased 38.3 and 1.8 times respectively than in 1999 (χ(2) = 304.025, P < 0.001; χ(2) = 42.845, P < 0.001) and other SH remained unchanged. (3) The prevalence of PA complicated with OSAS increased significantly in recent five years (χ(2) = 26.376, P < 0.001). Incidence of OSAS was 23.9% in PA patients and incidence of PA was 6.7% in OSAS patients. CONCLUSIONS: With the insights gained on hypertension mechanism and the development of new diagnostic technology, percent of diagnosed SH increased remarkably in recent years in hospitalized hypertensive patients in our department of hypertension. OSAS and PA are the leading causes of SH.

[Etiology analysis of 628 patients with refractory hypertension].

OBJECTIVE: To analyze the etiology of 628 patients with refractory hypertension and to observe the disease distribution with respect to gender and different age groups. METHODS: In this study, clinical data of 628 refractory hypertensives who hospitalized in our hospital from September 1997 to December 2005 were retrospectively analyzed. RESULTS: (1) There were 80.1% (503/628) patients with essential hypertension, 18.9% (119/628) with secondary hypertension (SH) while diagnosis was not clear in 1.0% (6/628) patients. Renovascular hypertension (33.6%) and obstructive sleep apnea syndrome (23.5%) were the major causes of SH. The highest prevalence rate of endocrine hypertension was primary aldosteronism (13.5%). (2) There were significantly more male patients than female patients with essential hypertension, SH, renal hypertension, obstructive sleep apnea syndrome, primary aldosteronism while the incidence of pheochromocytoma in female was significantly higher than that in male patients (all P < 0.05). The incidence of renovascular hypertension was similar between male and female patients. (3) SH occurred more often in young patients (33.1%) than in aged patients (13.8%, P < 0.05). CONCLUSION: Our data from this patient cohort showed that SH, especially renovascular hypertension and obstructive sleep apnea syndrome are major causes for refractory hypertension in young patients and primary aldosteronism was the commonest reason of endocrine hypertension in youth and middle-aged patients with refractory hypertension.

[The relationship between dyslipemia and the polymorphism of angiotensin II type 1 receptor gene in hypertensive Kazakans of Xinjiang.].

OBJECTIVE: To investigate the relationship between dyslipidemia and the polymorphism of angiotensin II type 1 receptor (AT(1)R) gene A1166C in hypertensive Kazakans of Xinjiang area. METHODS: Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) methods were used to detect the A1166C polymorphism of AT(1)R gene of 296 hypertensive and 198 normotensive Kazakans. Biochemical parameters were measured by autochemical emalyzer. RESULTS: (1) The TC and LDL-C levels are significantly higher in hypertension group than those in normotensive controls [TC: (4.91 +/- 1.19) mmol/L vs. (4.43 +/- 1.20) mmol/L; LDL-C: (3.36 +/- 1.01) mmol/L vs. (2.94 +/- 1.30) mmol/L, P < 0.001). (2) In hypertension group, TC and LDL-C are related to A1166C polymorphism of AT(1)R gene and TC and LDL-C of AC carriers are significantly higher than AA carriers (P < 0.05). CONCLUSION: The dyslipidemia is related to A1166C polymorphism of AT(1)R gene in hypertensive Kazakans.

[Analysis of etiology of the patients with hypertension from the People's Hospital of Xinjiang Uygur Autonomous Region].

OBJECTIVE: To analyze the etiology of the patients with hypertension from the People's Hospital of Xinjiang Uigur Autonomous Region, and to investigate the distribution of hypertension in gender and different ages. METHODS: From September 1997 to December 2005, the data of 4642 patients with hypertension was retrospective studied. RESULTS: (1) Of all the patients, 85.24% were essential hypertension (EH) and 14.76% were secondary hypertension (SH). Higher prevalence of sleep apnea syndrome (42.92%) and anxiety (15.04%) was found in secondary hypertension. The highest prevalence of primary aldosteronism (12.12%) was found in endocrine hypertension. (2) The male patients with hypertension were more than the female ones, and the incidence of EH, sleep apnea syndrome (SAS) and primary aldosteronism was higher in male patients than female ones, and the following was less than female: anxiety, pheochromocytoma and renovascular hypertension. (3) Among the patients with SH, 21.9% were found in youth, and 9.85% in aged. CONCLUSION: For the young, SH should be excluded, especially SAS and anxiety should be screened and differentiated. The highest prevalence of endocrine hypertension is primary aldosteronism in young and middle-aged male. The prevalence of pheochromocytoma in female is higher than that of male.

PKA activation bypasses the requirement for UNC-31 in the docking of dense core vesicles from C. elegans neurons.

The nematode C. elegans provides a powerful model system for exploring the molecular basis of synaptogenesis and neurotransmission. However, the lack of direct functional assays of release processes has largely prevented an in depth understanding of the mechanism of vesicular exocytosis and endocytosis in C. elegans. We address this technical limitation by developing direct electrophysiological assays, including membrane capacitance and amperometry measurements, in primary cultured C. elegans neurons. In addition, we have succeeded in monitoring the docking and fusion of single dense core vesicles (DCVs) employing total internal reflection fluorescence microscopy. With these approaches and mutant perturbation analysis, we provide direct evidence that UNC-31 is required for the docking of DCVs at the plasma membrane. Interestingly, the defect in DCV docking caused by UNC-31 mutation can be fully rescued by PKA activation. We also demonstrate that UNC-31 is required for UNC-13-mediated augmentation of DCV exocytosis.

Silence of synaptotagmin I in INS-1 cells inhibits fast exocytosis and fast endocytosis.

Synaptotagmin I (Syt I) is a Ca(2+) sensor for triggering fast synchronized release of neurotransmitters. However, controversy remains whether Syt I is also obligatory for the exocytosis and endocytosis of larger dense core vesicles (LDCVs) in endocrine cells. In this study, we used a short hairpin RNA (shRNA) to silence the expression of Syt I and investigated the roles of Syt I on exocytosis and endocytosis in INS-1 cells. Our results demonstrated that expression of Syt I is remarkably reduced by the Syt I gene targeting shRNA. Using high-time resolution capacitance measurement, we found that the silence of Syt I decreased the calcium sensitivity of fusion of insulin granules and therefore reduced the exocytotic burst triggered by step-like [Ca(2+)](i) elevation. In addition, the occurrence frequency and amplitude of fast endocytosis were remarkably reduced in the silenced cells. We conclude that Syt I not only participates in the Ca(2+)-sensing of LDCV fusion with plasmalemma, but also plays a crucial role in fast endocytosis in INS-1 cells.

Heterogeneity of the Ca2+ sensitivity of secretion in a pituitary gonadotrope cell line and its modulation by protein kinase C and Ca2+.

Modulation of the Ca2+ sensitivity and cooperativity of secretion is an important means of regulating neurotransmission and hormone secretion. Employing high-time resolution measurement of membrane capacitance (Cm) stimulated by step-like or ramp [Ca2+]i elevation, we have identified the co-existence of both a high and low Ca2+-sensitive exocytosis in an immortal pituitary gonadotrope cell line, LbetaT2. Ramp [Ca2+]i generated by slow uncaging elicited a biphasic C(m) response. The first phase of response, which represents a highly Ca2+-sensitive pool (HCSP) of vesicles, began to secrete at low [Ca2+]i concentration (<1 microM) with low Ca2+ cooperativity. In contrast, the second phase, which represents a lowly Ca2+-sensitive pool (LCSP) of vesicles, only exocytozed at higher [Ca2+]i (>5 microM) and displayed a steep Ca2+ cooperativity. The co-existence of vesicle populations with different Ca2+ sensitivities was further confirmed by flash photolysis stimuli. The size of the HCSP was approximately 30 fF under resting conditions, but was dramatically increased (approximately threefold) by application of phorbol-12-myristate-13-acetate (PMA, an activator of protein kinase C). Forskolin (an activator of protein kinase A), however, exerted no significant effect on the size of both HCSP and LCSP. GnRH (gonadotropin releasing hormone) augmented the size of both pools to a larger extent (5- and 1.7-fold increase for HCSP and LCSP, respectively). The heterogeneity of Ca2+ sensitivity from different pools of vesicles and its differential modulation by intracellular signals suggests that LbetaT2 cells are an ideal model to further unravel the mechanism underlying the modulation of Ca2+-sensing machineries for exocytosis.

[The relationship between the beta-2-adrenergic receptor gene +46 Arg 16/Gly variant and serum level of low density lipoprotein-cholesterol in Kazakans of Xinjiang].

OBJECTIVE: To investigate the distribution of beta-2-AR +46 A-->G variant in Kazakans of Xinjiang and the relationship of the variant with low density lipoprotein-cholesterol (LDL-C) level in this population. METHODS: The genotypes of beta-2-AR gene Arg16/Gly variant were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in 506 Kazakans with age from 30 to 69, and its distribution and relationship to LDL-C level were investigated. RESULTS: (1) The frequencies of genotypes AA, AG, GG and alleles A, G of beta-2-AR +46 variant in this population were 0.310, 0.455, 0.235 and 0.538, 0.462 respectively, which were accorded with Hardy-Weinberg equilibrium. (2) The Gly16/Gly genotype had highest LDL-C level in the three genotypes, and were significantly higher than Arg16/Gly genotype (P < 0.05). (3) Comparing the effect of beta-2-AR gene +46 variant on serum lipid in males with females, we found that females with Gly16/Gly genotype had the highest level of serum LDL-C. CONCLUSIONS: These data show that Gly16/Gly genotype of beta-2-AR gene +46 A-->G variant is associated with higher level of serum LDL-C in this population, especially in female.

[Construction of replication-deficient recombinant adenovirus expressing gag-polDelta and gp140TM genes of human immunodeficiency virus in mice].

BACKGROUND: Construction of replication-deficient recombinant adenovirus expressing gag-pol and env genes of human immunodeficiency virus (HIV) in mice. METHODS: gag-polDelta and gp140TM genes were cloned into shuttle vector pAdTrack-CMV respectively, and then the plasmids containing gag-polDelta or gp140TM gene were cotransformed with the backbone of adenovirus into E.coli BJ5183. Transfections of the recombinants were performed to obtain recombinant adenoviruses. Its immunogenicity was evaluated by testing antibody levels of mice primed with DNA vaccines and boosted with recombinant adenoviruses. RESULTS: The replication-deficient recombinant adenovirus could express Gp140TM, Gag P55 and P24 proteins correctly. The mice primed with DNA vaccines and boosted with recombinant adenoviruses elicited high titer of HIV-1-specific antibody compared with that inoculated with DNA vaccines only. CONCLUSION: Replication-deficient recombinant adenovirus expressing gag-polDelta and gp140TM can elicit high titer HIV-1-specific antibodies.

[The relationship between the variants in 5' upstream core promoter A(-6)G and A(-20)C of angiotensinogen gene and essential hypertension in Kazakans of Xinjiang].

OBJECTIVE: To investigate whether the variants A(-6)G and A(-20)C of angiotensinogen (AGT) gene are involved in the pathogenesis of essential hypertension in Kazakans. METHODS: T his case control study recruited 125 subjects with hypertension and 74 normotensive subjects from Kazakans of Xinjiang. Genomic DNA from leukocytes was analyzed for genetic variants A(-6)G and A(-20)C in 5' upstream core promoter of AGT gene by polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), restriction fragment length polymorphism (RFLP) and automatic sequencing. RESULTS: (1)There were only A(-6)G and A(-20)C variants in the -164 to +73 region of Kazakans' AGT gene. (2) The distributions of genotypes AA, AG, GG at locus -6 of AGT gene showed significant difference between the hypertensive group (0.39, 0.45, 0.16) and the normotensive group (0.49, 0.49, 0.02; Chi2=8.56, P=0.014). There were evident differences in the frequencies of the -6A and the -6G allele of the two groups (0.62, 0.38 and 0.73, 0.27; Chi2=5.35, P=0.021). (3) No significant difference was observed in the distribution of genotypes AA, AC, CC at locus -20 of AGT gene between the hypertensive group (0.69, 0.26, 0.05) and the normotensive group (0.65, 0.32, 0.03; Chi2=2.42, P=0.30). There was no distinct difference in the frequencies of the -20A allele and the -20C allele of the two groups (0.82, 0.18 and 0.82, 0.18; Chi2=0, P=0.99). (4) No significant difference was found at the levels of systolic and diastolic blood pressure between the groups corresponding to genotypes at the loci -6 and -20 of AGT gene. CONCLUSION: The results suggest that the polymorphism of A(-6)G in 5' upstream core promoter of the AGT gene may be involved in the pathogenesis of essential hypertension in Kazakans, while the A(-20)C variant may not play an important role in the etiology of essential hypertension in Kazakans.