OBJECTIVETo analyze the clinical value of 24-hour urinary sodium determination in children with postural tachycardia syndrome (POTS).

METHODFifty-eight POTS children and 10 healthy children (control group) from Peking University First Hospital during June 2012 to May 2014 were enrolled. Their 24-hour urinary sodium and plasma sodium levels were compared. Correlation analysis was done between 24-hour urinary sodium and symptom scores in children with POTS. All patients were treated with oral rehydration salts. The POTS patients were divided into hyponatriuria group (urinary sodium < 124 mmol/24 h) and hypernatriuria group (urinary sodium ≥ 124 mmol/24 h). Kaplan-Meier curve was used to analyze the effects of different 24-hour urinary sodium levels in children with POTS receiving rehydration salts therapy.

RESULTThe 24-hour urinary sodium levels of children with POTS were significantly lower than that of control group ((110. 0 ± 45. 8) vs. (221. 3 ± 103. 6) mmol/24 h, t =3. 339, P = 0. 008), while no statistical significance was found in plasma sodium between the two groups ((139. 7 ± 2. 1) vs. (139. 7 ± 2. 3) mmol/L, t = 0. 082, P = 0. 935). Pearson correlation analysis showed that 24-hour urinary sodium and severity of symptoms in children patients were negatively correlated (r = - 0. 654, P < 0. 001) . Urinary sodium < 124 mmol/24 h was used as the cut-off value, there were 43 cases in hyponatriuria group and 15 cases in hypernatriuria group. The symptom scores were significantly higher in hyponatriuria group (10. 2 ± 3. 7 vs. 5. 0 ± 1. 8, P < 0. 001), there was no significant difference in other basic information and hemodynamic data between groups (P > 0. 05). Logistic regression analysis revealed that urine sodium < 124 mmol/24 h was independent risk factor for effectiveness of rehydration salts in POTS patients (OR = 0. 043, 95% CI:0. 004 - 0. 499, P = 0. 012). Kaplan-Meier survival analysis showed the long-term effect of patients receiving oral rehydration salts in hyponatriuria group was significantly better than that in hypernatriuria group (86. 0 % vs. 60. 0%, χ2 = 8. 471, P = 0. 004).

CONCLUSIONTwenty-four hours urinary sodium is a good indicaor for guiding children with POTS receiving rehydration salts therapy.

Case-Control Studies ; Child ; Fluid Therapy ; Hemodynamics ; Humans ; Postural Orthostatic Tachycardia Syndrome ; urine ; Rehydration Solutions ; Salts ; Sodium ; urine

AIM:To investigate the effects of seipin gene deficiency on renal injury and the possible mecha-nisms in seipin-/-mice.METHODS:Six-month-old male seipin knockout ( seipin-/-) and wild-type ( WT) mice ( n=8) were used to study 24 h urinary albumin excretion ( UAE) , renal functions, pathological changes, and plasma leptin and adiponectin levels.Seipin mRNA expression in different tissues and each part of the kidney was also measured in WT mice.RESULTS:Real-time PCR analysis showed seipin mRNA expression in WT mice was higher in adipose tissue and testicles, and was also found in the kidney, which was mainly in glomeruli.Compared with control group, seipin-/-group showed increased kidney weight/tibia length (P<0.01), 24 h UAE (P<0.01), creatinine clearance (P<0.01), and glomerular and mesangial surface area (P<0.05).Both plasma leptin (P<0.01) and adiponectin (P<0.05) levels were significantly decreased in seipin-/-mice.CONCLUSION:Seipin gene deficiency in mice leads to renal injury prob-ably by decreasing plasma leptin and adiponectin levels due to lack of adipose tissue.

Objective To construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.

OBJECTIVEThe imbalance between extracellular matrix (ECM) synthesis and degradation induces the excessive ECM deposition and thus renal fibrosis. The decoction (A&A) which is a combination of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has been shown to alleviate ECM production in animal models of chronic kidney diseases. In this paper, the effect of A&A on ECM degradation was investigated with interstitial fibrosis in rats.

METHODMale Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After administration of A&A (14 g x kg(-1) x d(-1)) by gavage for 3, 7 and 10 days, morphological changes were evaluated by HE, PAS and Sirius red staining technique. The expression of plasminogen activator inhibitor-1 (PAI-1) and tissue-type plasminogen activator (t-PA), the activity of PAI-1 and t-PA were determined by ELISA. The activity of matrix metalloproteinases (MMP-9, MMP-2), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) were evaluated by gelatin zymography or reverse gelatin zymography, respectively.

RESULTMorphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which was reduced by A&A administration (P < 0.05, UAA vs UUO group). Compared with the sham group, the expression of PAI-1 was significantly increased in UUO group by 63%, 91% and 112% at day 3, 7 and 10 respectively; and there were a remarkable decrease in UAA group by 44%, 43% and 52% at day 3, 7 and 10. The expression of active PAI-1 was strikingly increased in UUO group at day 3 [(30.5 +/- 23.8) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], day 7 [(36.5 +/- 11.2) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], and day 10 [(54.5 +/- 14.2) ng x g(-1) vs. (0.5 +/- 0.5) ng x g(-1), P < 0.05)]. The active PAI-1 was decreased in UAA group at day 7 [(14.9 +/- 0.5) ng x g(-1) vs. (36.5 +/- 11.2) ng x g(-1), P < 0.05] and day 10 [(15.4 +/- 4.8) ng x g(-1) vs. (54.5 +/- 14.2) ng x g(-1), P < 0.05]. The expression of t-PA was increased in UUO group only at day 3 [(58.1 +/- 16.5) microg x g(-1) vs. (30.1 +/- 17.3) microg x g(-1)], P < 0.05), meanwhile decreased in UAA group [(26.3 +/- 8.7) microg x g(-1) vs. (58.1 +/- 16.5) microg x g(-1), P < 0.05)]. But the expression of active t-PA was shown no significantly difference among the three groups. For MMP-2 and MMP-9 activity, they were significantly higher compared with the sham group in UUO group, but no significantly change after A&A treatment. The TIMP-1 activity was significantly increased in UUO group by 28% and 63% at day 7 and 10 respectively, significantly decreased in UAA group by 40% and 39% at the same time point.

CONCLUSIONThe anti-fibrosis effects of A&A might be associated with modulating the imbalance of PAs/PAIs system as well as MMPs/TIMPs system, thereby alleviate ECM accumulation and interstitial fibrosis.

Angelica sinensis ; chemistry ; Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Extracellular Matrix ; metabolism ; Fibrosis ; Humans ; Kidney ; enzymology ; metabolism ; pathology ; Kidney Diseases ; drug therapy ; enzymology ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism ; Rats ; Rats, Wistar ; Tissue Plasminogen Activator ; metabolism

Objective To investigate the potential dosimetric advantages of half jaw volumetric modulated arc therapy ( H-VMAT) applied to the Oropharyngeal Cancer, comparing with full jaw VMAT (F-VMAT) and intensity modulated radiotherapy ( IMRT ). Methods Planning CT images of 10 oropharyngeal cancer patients were retrospectively chosen and transferred to Eclipse treatment planning system v. 11. 0 (Varian Medical Systems, Pala Alto, USA), based on which H-VMAT, W-VMAT, and IMRT plans were created. Two full arcs (360°) were adopted for VMAT planning, and the 7 beams were equally distributed for IMRT planning. The optimization constraints remained the same for the three kinds of plans. The dosimetric parameters such as D2 , D98 , D50 , HI, and CI were evaluated for PGTV, PCTV1, PCTV2, PGTVln, and PCTVln. In addition, the maximum dose (Dmax) and D1 cc(minimum dose received by 1cc) of the brainstem and spinal cord were analyzed respectively. The mean dose ( Dmean ) to the parotids, oral cave, larynx, and cervical normal tissues were also reviewed. The monitor units ( MU) for all treatment plans were recorded. Results Comparisons of the three planning techniques showed that H-VAMT improved the HI and CI of the targets (except PCTV2) significantly (HI: F =3. 959, 6. 764, 10. 581, 6. 770, 13. 040, P<0. 05;CI:F=6. 594, 4. 138, 0. 842, 4. 031, 5. 388, P<0. 05);reduced Dmax(F=4. 509, 20. 331, P<0. 05) and D1 cc for brainstem and spinal cord (F=27. 432, 26. 314, P<0. 05) significantly;reduced Dmean(F=4. 279, 29. 498, 19. 295, P<0. 05) to the normal tissues of the mouth, throat and neck significantly. The V50 of the mouth and throat were slightly lower in IMRT plans (F=8. 140, P<0. 05). IMRT was slightly better than W-VMAT in sparing oral cavity and larynx, but the dose distribution was the worst. The H-VMAT plans showed the best dose distribution in the cervical normal tissues, especially for the lower and posterior parts, where IMRT plans displayed high dose curves. Conclusions H-VMAT is dosimetrically superior than W-VMAT and IMRT for oropharyngeal cancer, which could be considered for clinical applications.

OBJECTIVETo study the promoter methylation status of SFRP genes and the effect of 5- aza- 2'- deoxycytidine (5- Aza- CdR)induced apoptosis via Wnt/β- catenin pathway by demethylation in Jurkat cells.

METHODSJurkat cells were treated with different concentrations of 5- Aza- CdR. The cell proliferation level of Jurkat cells was detected by MTT assay. Apoptosis was evaluated by flow cytometry. Methylation- spcific PCR (MSP) was used to determine the methylation status of SFRP genes. The expressions of SFRP genes were detected by real time fluorescence quantitative PCR. The mRNA expression levels of survivin, c- myc and cyclin- D1 were analyzed by RT- PCR. Western blot was used to detect the levels of β-catenin protein.

RESULTSCompared with control group, the different concentrations of 5-Aza-CdR could significantly inhibit the proliferation of Jurkat cells in a time-dose dependent manner (P<0.05). After being treated by 5- Aza- CdR for 48 hours, the cell early apoptosis rate in experiment group was significantly higher than that in control group (P<0.05). The promoters of SFRP1, SFRP2, SFRP4, SFRP5 genes were hypermethylation state in the control group, after being treated by 5-Aza-CdR for 72 hours, the brightness of SFRP1, SFRP2, SFRP4, SFRP5 genes' methylation strips weakened in a dose- dependent manner. SFRP mRNA expression increased (P<0.05) when 5- Aza- CdR concentration increased, and the level of β- catenin protein was dampened in a dose- dependent manner (P<0.05). As compared to the control group, the mRNA expressions of associated apoptosis genes survivin, c-myc and cyclin- D1, respectively were obviously down- regulated in a dose- dependent manner (P<0.05).

CONCLUSIONThe effect of demethylation could up- regulate SFRP genes expressions by reversing its hypermethylation and induced apoptosis by down-regulation of β-catenin and associated apoptosis genes.

Apoptosis ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Proliferation ; DNA Methylation ; Down-Regulation ; Gene Expression ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jurkat Cells ; Membrane Proteins ; genetics ; Promoter Regions, Genetic ; Wnt Signaling Pathway ; beta Catenin ; metabolism

Objective To explore the characters of lower urinary tract symptoms (LUTS) in patients with Parkinson's Disease (PD).Methods From Oct 2013 to Jun 2016,after evaluating of movement disorder by modified Hoehn-Yahr(H-Y) scale and LUTS by international prostate symptom score (IPSS),urodynamic study was performed in PD patients with LUTS.The incidence of every symptom of LUTS,the relationships between the IPSS categories and urodynamic study were analyzed.Results 64 patients (containing 26 male and 38 female) with 40-80 (62.7 ± 10.2) years old were included.2,4,30,19,12 and 6 patients were belonged to modified H-Y scale 1-4,respectively.Frequency (50 patients,78.1％) was the most common LUTS,while frequency all day (20 patients,31.3％) was the most common symptom for the most severe LUTS.IPSS was 1 7.5 ± 7.8 (4-35) and quality of life was 5.1 ± 0.6 (4-6) for the patients.There was no significant correlation between modified H-Y scale and IPSS (P ＞ 0.05).According to the criteria of IPSS,28 patients (43.8％) only had irritative symptoms,3 patients (4.7％) only had obstructive symptoms,while 26 patients (40.6％) had mixed symptoms and 7 patients (10.9％) belonged to no one.Urodynamic study showed 11 patients (39.3％) with only irritative symptoms had detrusor overactivity(DO),6 patients (23.1％) with mixed symptoms had DO + bladder outlet obstruction (BOO) or DO + detrusor underactivity,however,there was no one with BOO in the three patients with only obstructive symptoms.Conclusion Frequency was the most common LUTS,while frequency all day was the most common symptom for the most severe LUTS in PD patients.Irritative and mixed symptoms were common in PD patients with LUTS,but the urodynamic results were not consistent with the category of LUTS in most of the patients.LUTS severity was not correlated with movement disorders in PD patients.

Objective To investigate the influences of herpes simplex virus 1 and 2 ( HSV1 and HSV2) infection on the expression of signaling molecules associated with innate immune response in respira-tory and vaginal epithelial cells for bettering understanding of HSV infection and pathological characteristics in the primary infection site, namely mucosal epithelial tissues. Methods KMB17 and VK2 cells were in-fected with HSV. Changes in cell morphology and inner structure after HSV infection were observed under optical microscope and scanning electron microscope, respectively. Viral proliferation in KMB17 and VK2 cells was detected by plaque assay, microcytopathic assay and real-time quantitative PCR. Expression of sig-naling molecules associated with innate immune response in virus-infected KMB17 and VK2 cells were ana-lyzed by real-time quantitative PCR. Results Both HSV1 and HSV2 could infect KMB17 and VK2 cells, and cause damage to cell morphology and inner structure after 12 hours. Both of the two viruses formed simi-lar plaque on the single layer of KMB17 and VK2 cells, although HSV2 proliferated slower than HSV1. There were differences in the expression of signaling molecules associated with innate immune response in-duced by the two viruses in KMB17 and VK2 cells. Conclusion Both HSV1 and HSV2 could infect and proliferate in epithelial cells ( KMB17 and VK2 cells) . Although there were slight differences in viral prolif-eration between them, significant differences in the expression of signaling molecules associated with innate immune response induced by the two viruses were observed.

Candida albicans is the most abundant fungal species in oral cavity. As a smart opportunistic pathogen, it increases the virulence by switching its forms from yeasts to hyphae and becomes the major pathogenic agent for oral candidiasis. However, the overuse of current clinical antifungals and lack of new types of drugs highlight the challenges in the antifungal treatments because of the drug resistance and side effects. Anti-virulence strategy is proved as a practical way to develop new types of anti-infective drugs. Here, seven artemisinins, including artemisinin, dihydroartemisinin, artemisinic acid, dihydroartemisinic acid, artesunate, artemether and arteether, were employed to target at the hyphal development, the most important virulence factor of C. albicans. Artemisinins failed to affect the growth, but significantly inhibited the hyphal development of C. albicans, including the clinical azole resistant isolates, and reduced their damage to oral epithelial cells, while arteether showed the strongest activities. The transcriptome suggested that arteether could affect the energy metabolism of C. albicans. Seven artemisinins were then proved to significantly inhibit the productions of ATP and cAMP, while reduced the hyphal inhibition on RAS1 overexpression strain indicating that artemisinins regulated the Ras1-cAMP-Efg1 pathway to inhibit the hyphal development. Importantly, arteether significantly inhibited the fungal burden and infections with no systemic toxicity in the murine oropharyngeal candidiasis models in vivo caused by both fluconazole sensitive and resistant strains. Our results for the first time indicated that artemisinins can be potential antifungal compounds against C. albicans infections by targeting at its hyphal development.
Animals ; Mice ; Candida albicans ; Candidiasis, Oral/drug therapy* ; Antifungal Agents/pharmacology* ; Hyphae ; Artemisinins/pharmacology*