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OBJECTIVE To assess the association of SLC12A3 and SCNN1B gene polymorphisms (rs11643718 and rs12447134) with essential hypertension among ethnic Koreans from Mudanjiang, China. METHODS For 204 patients with essential hypertension and 186 healthy controls, the genotypes of rs11643718 and rs12447134 loci were determined with an improved multiplex ligase detection reaction (iMLDR) method. RESULTS Allelic and genotypic frequencies of rs11643718 of SLC12A3 gene are associated with the onset of disease hypertension (P <0.05) as well as systolic blood pressure (P < 0.01, under a recessive model). No association was found between rs12447134 of SCNN1B gene with the onset of disease (P > 0.05) but diastolic blood pressure (P < 0.05, under a recessive model). CONCLUSION The polymorphisms of rs11643718 locus is associated with the susceptibility for essential hypertension among ethnic Koreans from Mudanjiang area and can be used as a predictor for the disease.
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Objective@#To investigate the interaction of Ca2+ protein TRPC1 and STIM1 in extracellular Ca2+ -sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO).@*Methods@#Human umbilical vein endothelial cells (HUVECs) were cultured and incubated with CaR agonist spermine (activating store-operates cation channels (SOC) and receptor-operated channels (ROC)), CaR negative allosteric modulator Calhex231 (blocking SOC, activating ROC) and ROC analogue TPA (activating ROC, blocking SOC), protein kinase C (PKC) inhibitor Ro31-8220, PKCs and PKCμ inhibitor Go6967(activate SOC, blocking ROC), respectively. The interaction of TRPC1 and STIM1 was determined using the immunofluorescence methods. The interaction between TRPC1 and STIM1 were examined by Co-immuno precipitation. The HUVECs were divided into: TRPC1 and STIM1 short hairpin RNA group (shTRPC1+ shSTIM1 group), vehicle-TRPC1+ vehicle-STIM1 group and control group. The cells were incubated with four different treatments under the action of above mentioned interventions, intracellular Ca2+ concentration ([Ca2+ ]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, the production of NO was determined by DAF-FM.@*Results@#(1) The expression of TRPC1 and STIM1 proteins levels in HUVECs: Under the confocal microscope, TRPC1 and STIM1 protein expression showed masculine gender, both located in cytoplasm in the normal control group. Post incubation with Calhex231+ TPA, Ro31-8220 and Go6967, TRPC1 and STIM1 positioned in cytoplasm was significantly reduced, and the combined TRPC1 and STIM1 was also significantly reduced. (2) The interaction of TRPC1 and STIM1 in HUVECs: The relative ratios of Calhex231+ TPA+ Spermine+ Ca2+ group, Ro31-8220+ Spermine+ Ca2+ group and Go6976+ Spermine+ Ca2+ group STIM1/TRPC1 and TRPC1/STIM1 were as follows: (25.98±2.17)% and (44.10±4.01)%, (20.85±1.01)% and (46.31±3.47)%, (23.88±2.05)% and (39.65±2.91)%, which were significantly lower than those in the control group (100.00±4.66)% and (100.00±6.40)% and in the Spermine+ Ca2+ group (106.04±2.45)% and (107.78±2.66)% (all P<0.05). (3) The influence of joint TRPC1 and STIM1 transfection to four different drugs treated HUVECs on [Ca2+ ]i and NO generation: The changes of two excitation fluorescence intensity ratio and NO net fluorescence intensity values were consistent, [Ca2+ ]i and NO net fluorescence intensity values were significantly lower in the experimental group than the control group and the vehicle group (all P<0.05), while which were similar between the vehicle group and control group (all P>0.05).@*Conclusions@#Our results indicate that TRPC1 and STIM1 jointly regulate CaR-mediated Ca2+ influx and nitric oxide generation in HUVECs in the form of binary complex.
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Objective To explore a surgical method for the treatment of lumbosacral spinal tuberculosis by combination of one-stage focus debridement with anterolateral incision, bone graft fusion with titanium mesh cage and internal fixation with double pedicle crew system.Methods From Sep.2009 to Dec.2012, a total of 8 patients with lumbosacral spinal tuberculosis which included 5 cases of male, 3 cases of female.The age ranged from 20 to 65 years, with a mean of 51.6 years.All patients presented with persistent back pain, 4 patients with radiating pain of unilateral lower limb, 3 with weakness and numbness and 5 with constitutional symptoms including low-grade fever and weight loss.All patients were not associated with active tuberculosis in oth er parts of the body.The patients were given regular anti-TB treatment for at least 4 weeks.By anterolateral incision, common iliac and iliac arteries and veins were dissociated extraperitoneally.The focus was completely debrided through the inferior part of vessels.Then the bone graft fusion was performed with the titanium mesh cage and the internal fixation with a double pedicle crew system was accomplished.After the surgery, patients were treated with continuous anti-TB drugs and with antibiotics to prevent infection.Patients were allowed to move with the protection of waist early and regular follow-up.Results Operation time was 180-360 min, with an average of 225 min.Operative blood loss was 624 ml and drainage volume was 150 ml on average.All cases were cured after surgery.No severe complications were observed during the surgeries.After follow-up of 8 to 30 months (averaged 12months), no recurrence of the tuberculosis was found.The lumbocrural pain improved in all the patients.Complications such as migration, loosening and breaking of the implants were not observed.The vertebral bodies were fused in all patients with an average time of 8.3 months.No case occurred angiemphraxis or internal bleeding.Conclusion The method debrids the focus of lumbosacral spinal tuberculosis thoroughly and implements titanium mesh cage and double pedicle crew system simultaneously.The pedicle screw system is implemented in anterior lumbosacral vertebrae through the inferior part of iliac arteries and veins, which will not lead to angiemphraxis or vascular injuries.The early term outcome is encouraging.This technique is safe and effective to treat severe lumbosacral spinal tuberculosis.
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Objective:To explore the regulation of prokaryotic ubiquitin-like protein ( Pup )-proteasome system on the persistence of Mycobacterium tuberculosis by inducing Mycobacterium tuberculosis to being persistence state under hypoxia conditions and then analyzing the difference on the expression levels of Pup,Dop,PafA and Mpa gene at various time and different conditions. Methods:The total mRNA of the international standard virulent strains of Mycobacterium tuberculosis(H37Rv),which were cultured under hypoxia and aerobic conditions, were extracted from each group at various time and purity of the mRNA were identified.The expression of Pup,Dop,PafA and Mpa genes of M.tuberculosis strains in each group were quantified by SYBR Green I qRT-PCR,which aimed at finding the difference among the expression of Pup,Dop,PafA and Mpa genes at various time and different conditions.Results:The expression levels of Pup, Dop, PafA and Mpa genes in Mycobacterium tuberculosis under hypoxia conditions were measured at various times.The expression levels of Pup,Dop,PafA and Mpa genes:compared with the 0 d,the expression of the Pup gene was up-regulated 1.66,2.43 and 2.76-fold at 4 d,7 d,10 d respectively(P<0.05);the expression of the Dop gene was up-regulated 1.38, 1.91,2.54 and 3.28-fold at 2 d,4 d,7 d,10 d respectively(P<0.05);the expression of the PafA gene was up-regulated 1.22,1.75, 2.37 and 2.67-fold at 2 d,4 d,7 d,10 d respectively( P<0.05);the expression of the Mpa gene was up-regulated 1.66,2.21 and 2.63-fold at 4 d,7 d,10 d respectively(P<0.05).Take the aerobic conditions as control,under hypoxic conditions with the same culture time,the expression of the Pup gene was up-regulated 1.85,2.81 and 2.93-fold in 4 d,7 d,10 d respectively(P<0.05);the ex-pression of the Dop gene was up-regulated 1.20,1.76,2.01 and 3.01-fold in 2 d,4 d,7 d,10 d,respectively( P<0.05);the expression of the PafA gene was up-regulated 1.22,1.57,2.29 and 2.29-fold in 2 d,4 d,7 d,10 d,respectively(P<0.05);the expression of the Mpa gene was up-regulated 1.16,1.58,2.16 and 2.69-fold in 2 d,4 d,7 d,10 d,respectively(P<0.05).Conclusion:Under hypoxic conditions,there were significant differences on the expressions of Pup, Dop, PafA and Mpa genes at various times;what′s more, significant differences on the expressions of Pup,Dop,PafA and Mpa genes exist between hypoxic and aerobic conditions at the same time,Prokaryotic ubiquitin-like protein( Pup)-proteasome system plays a regulatory role on M.tuberculosis′s persistence.
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Objective:To discuss the change of ferritin ( Fn) and ferroportin expression quantity and time-related feature in the alveolar macrophages of mice , infected with different virulence of Mycobacterium Tuberculosis infected .Methods:The prepared bacte-ria of H37Rv or BCG were injected intravenously into the mice tails .On the day 1, 3, 5, 7, 9, 11, 13 and 15, the lavage fluids were collected and the alveolar macrophages were obtained from each group of mice .The expression of FPN and Fn were detected with ELISA and /or Western blot analysis .Results:The expression of Fn in the group of either H 37Rv or BCG infected mice was decreased on the day 7, 9 and 11, and was lowest on the day 7, which showed significantly statistical difference compared to that on the other days (P<0.05).The expression of FNP in the infected mouse macrophage was decreased gradually , which was obvious on the day 5. The expression levels reached to the lowest on the day 7 and 9.The expression was much lower than that in the negative control group (P<0.05).Conclusion:The expression of Fn and FPN in macrophages isolated from lungs of mice infected with Mycobacterium tu -berculosis H37Rv or BCG become decreased , and there is no difference between these two infected mouse groups .
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Objective:To explore the correlation between Mycobacterium tuberculosis PhoPR two-component system and the pathogenicity of different virulent MTB by analysing the expression levels difference of PhoP gene and PhoR gene in BCG ,H37Ra, H37Rv and XJ-MTB respectively.Methods:Total RNA extracted from four different virulent MTB strains and the integrity of total RNA were identified by using agarose gel electrophoresis.The expression of PhoP gene and PhoR gene were quantified by using SYBR Green I FQ-PCR.The expression levels difference of these genes were compared in different virulent MTB strains .Results: The relative expression levels of PhoP gene in between four different virulent MTB strains from high to low were XJ -MTB(9.05),H37Rv(1.00), H37Ra(0.25),BCG(0.08) respectively ,and the expression levels difference of PhoP gene were statistically significant in different virulent MTB strains ( P＜0.05 );the relative expression levels of PhoR gene in four different virulent MTB strains from high to low were XJ-MTB(5.72),H37Rv(1.00),H37Ra(0.18),BCG(0.07) respectively,and the expression levels difference of PhoR gene were sta-tistically significant in different virulent MTB strains ( P＜0.05 ).The expression levels of PhoP gene and PhoR gene at XJ-MTB were statistically significant difference compared with BCG ,H37Rv,H37Ra respectively (P＜0.05);the expression levels of PhoP gene and PhoR gene at H37Rv were statistically significant difference compared with BCG ,H37Ra respectively (P＜0.05);the expression levels of PhoP and PhoR gene at BCG were not statistically significant difference compared with H 37Ra (P>0.05).Conclusion:Significant expression levels difference of PhoP gene and PhoR gene are confirmed in different virulent MTB strains .Therefore,the Mycobacterium tuberculosis PhoPR two-component system is correlated with the pathogenicity in different virulent MTB strains.
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Objective To explore the correlation between Mycobacterium tuberculosis ( MTB ) PhoPR two-component system and drug resistance of MTB clinical isolates widespread in Xinjiang region by analyzing the expression of PhoP gene and PhoR gene among different isolates .Methods Total RNA of MTB was extracted from drug-susceptible strains , the strains only resistant to a single first-line anti-TB drugs (INH, RFP, SM and EB) and multidrug-resistant (MDR) strains, respectively.The purity of total RNA was checked by agarose gel electrophoresis .The expressions of PhoP gene and PhoR gene were quantified by using SYBR Green I qRT-PCR and the differences of their gene expression in different isolates were ana-lyzed.Results Compared with the drug-susceptible strains of MTB, the expression of PhoP gene was up-regulated for about 1.48 times in MTB strains resistant to RFP (RFP-MTB) and 2.74 times in MDR strain (P<0.05).Compared with MDR strain, the expressions of PhoP gene in the isolates resistant to INH (IN-HMTB), RFP (RFP-MTB), SM (SM-MTB) and EB (EB-MTB) were down-regulated for 0.70, 0.50, 0.25 and 0.21 times respectively.The expressions of PhoR gene were down-regulated for 0.36, 0.54, 0.35 and 0.19 times, respectively (P<0.05).The expressions of PhoR gene in the isolates of INH-MTB, RFP-MTB, SM-MTB and EB-MTB were up-regulated for 6.33, 4.56, 2.34, 1.85 and 9.06 times, respectively as compared with the drug-susceptible strains (P<0.05).Conclusion Significant differences of PhoR gene and PhoP gene expressions were observed among drug-susceptible strains , INH-MTB, RFP-MTB, SM-MTB, EB-MTB and MDR strains.Therefore, the Mycobacterium tuberculosis PhoPR two-component system is asso-ciated with the drug resistance of MTB strains prevalent in Xinjiang region .