Hypertonic Group Research Articles

Comparison of Mean Hospital Stay after Nebulization with 3% Hypertonic Saline vs Salbutamol in Treatment of Bronchiolitis

Abstract: Background: With an estimated prevalence of 33% in children age >2years and owing to 1 in 10th cause of hospitalization in children, Bronchiolitis is a major factor for morbidity and mortality. Varying nebulization doses have been shown to produce different clinical outcomes in patients with bronchiolitis. Objective: To compare mean hospital stay after nebulization with 3% hypertonic saline compared to the salbutamol among patients of bronchiolitis Materials and Methods: This was a randomized control study done at pediatric medicine ward, Services Hospital, Lahore. The study was done during 8th May 2018 to 8th November 2018. Data was collected from 68 children of age >3 months to < 2 years of both genders i.e. (34 in each group) fulfilling inclusion criteria. SPSS software was used for data entry and analysis. For quantitative variables age, mean and standard deviation were used whereas for qualitative variables like gender, frequency and percentages were used. Independent sample t-test was used for comparing mean hospital stay among the two groups taking p-value<0.05 as significant. Result: Average age in Salbutamol group was 1.06 ± 0.50 years and in Hypertonic solution mean age was 1.24 ± 0.55 years. There was significant difference in mean hospital stay with relatively lesser in hypertonic solution vs the Salbutamol group [3.18 ± 1.11 vs 4.44 ± 1.08 ; p-value <0.001]. Upon post-stratification analysis, the mean hospital stay was found to be significantly lower in the Hypertonic group vs. Salbutamol group with respect to age, gender and the duration of the disease (p-values<0.05 for all). Conclusion: This study concludes that the mean hospital stay was statistically lower in the hypertonic solution group vs the Salbutamol group. Moreover, the clinical efficacy of the hypertonic saline can be utilized in future to gain early recovery of Bronchiolitis and hence it can reduce the hospital stay and related cost. Keywords: Bronchiolitis, Salbutamol, Hypertonic saline, Hospital stay, Severity, Duration, Treatment.

Astragaloside IV Inhibited Podocyte Pyroptosis in Diabetic Kidney Disease by Regulating SIRT6/HIF-1α Axis.

To investigate the effect of astragaloside IV (AS) on podocytes pyroptosis in diabetic kidney disease (DKD). Forty male Sprague-Dawley rats were randomly divided into normal group (n = 10) and model group (n = 30). Rats in model group were intraperitoneally injected streptozotocin (60 mg/kg) for 3 days to induce DKD. Then rats were divided into DKD group, AS group, and UBCS group. The AS group was given 40 mg/kg/d of AS by gavage, and UBCS group was given 50 mg/kg/d of UBCS039 by gavage, and normal group and DKD group were given the same amount saline for 8 weeks, once a day. Hematoxylin-eosin and masson staining were used to observe pathology of kidney. Rat podocytes were divided into normal group, mannitol hypertonic group, high-glucose group, UBCS group, OSS group, and AS group. Western blotting, quantitative real-time polymerase chain reaction, immunofluorescence, and flow cytometry were used to analyze pyroptosis-related markers and reactive oxygen species (ROS) levels. Results showed that AS inhibited ROS and alleviated podocytes pyroptosis in rats by increasing expression of sirtuin 6 (SIRT6) and decreasing expression of hypoxia inducible factor 1 subunit alpha (HIF-1α). UBCS039 and AS enhanced SIRT6 level, decreased HIF-1α level, and finally improved pyroptosis of podocytes in vitro, whereas OSS-128167 showed the opposite effect for podocytes pyroptosis. AS improved podocytes pyroptosis in DKD by regulating SIRT6/HIF-1α pathway, thereby alleviating injury of DKD.

Comparison of Hypertonic Saline vs Salbutamol in Normal Saline Nebulization in Children with Bronchiolitis

Background: Main stay of treatment of acute Bronchiolitis is general supportive care, such as oxygenation, aspiration of respiratory tract secretions, and hydration therapy. Recent research indicates that breathed hypertonic saline is a potential treatment. Objective: To compare the mean length of hospital stay and clinical bronchiolitis score of salbutamol nebulized in normal saline versus hypertonic saline in children with acute bronchiolitis. Material and methods Study Design: Randomized controlled design Setting: Department of pediatrics, Fatima Memorial Hospital, Lahore. Duration of Study: 6 months after approval of the study i.e. from 12th April 2017 to 12th October 2017. Data Collection Procedure: Relevant parameters were identified, and the CBSS was computed. Using random tables, cases were divided among two groups and given salbutamol or hypertonic saline nebs. The length of stay was determined. The Clinical Bronchiolitis Severity Score (CBSS) was recorded based on various variables. Version 17 of the statistical package for social sciences was used for statistical analysis. Result: Patients in the salbutamol group had a mean age of 13.56.66 months, while those in the hypertonic saline group had a mean age of 12.46.58 months. In the group receiving salbutamol, there were 29 males and 31 females. There were 32 men and 28 women in the hypertonic saline group. After 24 hours, the mean CBSS in the salbutamol group was 6.132.47, whereas in the hypertonic saline group, it was 4.881.75. After 24 hours of treatment, the difference in CBSS was significant between the two groups (P0.05). The mean duration of hospitalization was 3.920.81 days with salbutamol and 3.050.77 days with the hypertonic group. In both groups, there was a marked difference in duration of hospital stay (P0.05). Conclusion: Thus, hypertonic saline was more effective than salbutamol in reducing the CBSS score and duration of hospital stay. Keywords: Hospital Stay, Clinical Bronchiolitis Score, Nebulized Salbutamol, Normal Saline, Hypertonic Saline, Children, Acute Bronchiolitis.

POS-352 MiR-26a-5p PROTECTS PODOCYTE FROM APOPTOSIS VIA DOWNREGULATION OF TRPC6 IN TYPE 2 DIABETIC NEPHROPATHY

Podocyte injury is a major etiological factor in diabetic nephropathy (DN), while transient receptor potential cationic channel 6 protein (TRPC6) channels play a key role in podocyte injury. Our study is designed to investigate the regulatory effect of microRNA-26a-5p (miR-26a-5p) on DN podocyte injury induced by TRPC6 and the underlying mechanisms. 20 SPF, 8-week male Sprague-Dawley (SD) rats were used for establishing the type 2 diabetic mellitus (T2DM) group by feeding them with high fat and high sugar food for 8 weeks and intraperitoneal injection of low-dose streptozotocin (STZ, 30mg/kg). Another 10 normal male SD rats were selected as normal control group (NC). The 24h urinary albumin excretion rate (UAE), urinary creatinine (Ucr), creatinine clearance rate (CCr) and other major biochemical parameters including the fasting insulin level (FINS), serum creatinine (Scr), fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL) were measured in both groups after establishing the DM models. Light and electron microscope were used to observe the pathology of kidneys and podocyte ultrastructures, respectively. Immunohistochemical method was used to show the distribution of nephrin and TRPC6 protein. To determine the expression of nephrin, TRPC6, apoptotic-associated proteins (cleaved-caspase-3 and bax/bcl-2), western blot was applied. qRT-PCR was used to detect miR-26a-5p and mRNAs of nephrin, TRPC6, cleaved-caspase-3 and bax/bcl-2. Podocyte apoptotic rate was measured using TUNEL method. Cultured immortalized mouse podocytes (MPC5) were divided into three groups: normal glucose group (NG), hypertonic group (HM), high glucose group (HG). Before and after miR-26a-5p mimics and inhibitor transfecting, the expression and mRNAs of nephrin, TRPC6, cleaved-caspase-3 and bax/bcl-2 were detected again using the same methods. Flow cytometry (FCM) was uesd to detect podocyte apoptosis. 1. Compared with NC group, rats in DM group exhibited disrupted biochemical conditions, for uAER, CCr, FINS, FBG, TC, TG and LDL were all significantly increased (all P <0.05) while HDL were decreased (P <0.05), along with significant insulin resistance. 2. Increased glomeruli volume, mesangial cell proliferation, thickened basement membrane and foot process fusion could be observed by light and electron microscope. 3. Immunochemistry results showed a increased expression of TRPC6 and a decreased of nephrin in DM rats than those in NC group (both P <0.05). 4. Both qRT-PCR and Western blot results in vivo and in vitro showed a notable increase in the expression of TRPC6, cleaved-caspase-3 and bax/bcl-2 and decrease in the expression of nephrin and miR-26a-5p (P <0.05). TUNEL and flow cytometry indicated higher podocyte apoptosis in DM rats and HG group. 5. The expression of TRPC6, cleaved-caspase-3 and bax/bcl-2 were down-regulated and nephrin expression was restored after miR-26a-5p mimics transfection (all P <0.05), while miR-26a-5p inhibitor intervention could aggravate all above changes as the effects of HG on podocytes (all P <0.05). High-glucose condition significantly decreased miR-26a-5p expression and increased TRPC6 expression and podocyte apoptosis. In DN, miR-26a-5p can protect podocyte from apoptosis via down-regulating the expression of TRPC6.

Identification of miR-145 as a regulator of the cardiomyocyte inflammatory response and oxidative stress under hyperglycemia.

The current study aimed to explore the effects of microRNA (miR)-145 on the inflammatory response and oxidative stress (OS) in high glucose (HG)-induced cardiomyocytes, as well as the specific mechanism underlying this action. H9c2 cells were treated with 33 mmol/l glucose (HG group) or cotreated with 24.5 mmol/l mannitol and 5.5 mmol/l glucose (hypertonic group), and the expression levels of miR-145 and ADP ribosylation factor 6 (ARF6) were detected. The cells were transfected with pcDNA3.1-ARF6, miR-145 mimics or corresponding negative controls prior to the assessment of cell survival rate. Levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS) and malondialdehyde (MDA), as well as the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), and the levels of IL-6, TNF-α and monocyte chemoattractant protein-1 (MCP-1) were subsequently determined. The apoptotic rate of H9c2 cells was examined by flow cytometry. The interaction between miR-145-ARF6 was predicted and confirmed by luciferase reporter assays. In the HG group, miR-145 expression was significantly decreased and ARF6 expression significantly increased compared with controls. Furthermore, the levels of inflammatory factors (IL-6, TNF-α and MCP-1), LDH, ROS and MDA were significantly elevated in the HG group compared with controls. Significantly decreased SOD, CAT and GPx activities and significantly increased numbers of apoptotic cells were observed in the HG group compared with controls. The cells transfected with miR-145 mimics exhibited significantly decreased LDH, ROS and MDA levels, significantly increased antioxidant enzyme activities and significantly decreased apoptotic rates compared with controls, while the opposite results were observed in cells transfected with pcDNA3.1-ARF6. Moreover, co-transfection with miR-145 mimics and pcDNA3.1-ARF6 exacerbated the inflammatory response and OS injury in HG-induced cardiomyocytes compared with cells transfected with miR-145 mimics alone. Furthermore, miR-145 negatively targeted ARF6. miR-145 attenuated the HG-induced inflammatory response and OS injury in cardiomyocytes by negatively regulating ARF6, which may contribute to providing a theoretical basis for the treatment of diabetic cardiomyopathy.

Percutaneous Endoscopic Gastrostomy and Nutritional Interventions by the Pediatric Nutritional Support Team Improve the Nutritional Status of Neurologically Impaired Children.

Aim: To evaluate the long-term effects of nutritional improvement following percutaneous endoscopic gastrostomy (PEG) tube feeding stratified by previous feeding method and to assess the impact of underlying muscle tone on the outcomes of the nutritional intervention. Methods: Neurologically impaired children who underwent PEG tube insertion and nutritional intervention provided by a pediatric nutritional support team were enrolled. We measured anthropometric variables, laboratory parameters, and nutritional intake at baseline, 6 months after PEG insertion, and the last visit. We evaluated the percent ideal body weight (PIBW), body mass index (BMI)-for-age z-score, and percentiles and calculated the ratios of calorie intake compared to required requirement (CIR) and protein intake compared to recommended requirement (PIR). Results: The PIBW and BMI-for-age z-score improved during the first 6 months (p = 0.003 and p = 0.005, respectively). The CIR (p = 0.015) and PIR (p = 0.004) increased during the study period. The baseline BMI and PIBW of the previous nasogastric tube feeding group were better than those of the oral feeding group (p = 0.02 and p = 0.03, respectively). The BMI-for-age z-score, PIBW, CIR, and PIR improved in the hypertonic group (p = 0.03, 0.02, 0.03, and 0.01, respectively). Conclusion: PEG tube feeding and active nutritional intervention improved the nutritional status of neurologically impaired children immediately after PEG insertion. The nutritional requirements might vary by the muscle tonicity.

COMPARATIVE STUDY TO ASSESS THE EFFECTIVENESS OF NEBULIZED 3% HYPERTONIC SALINE AND NEBULIZED LEVOSALBUTAMOL IN THE MANAGEMENT OF BRONCHIOLITIS

Objective: The objective of the study was to compare the effects of nebulized 3% hypertonic saline and nebulized levosalbutamol in the management of bronchiolitis.&#x0D; Methods: Seventy children of age 1–24 months admitted into the hospital with the diagnosis of bronchiolitis. Participants were divided into two groups of 35 each. Group A was given nebulized 3% hypertonic saline and Group B was given nebulized levosalbutamol. Modified respiratory distress assessment instrument (RDAI) is used at admission, at 48 h after admission, and at the time of discharge to identify the reduction in scores between two groups.&#x0D; Results: The mean age of patients in the study population was 10.1±6.4 months. The mean birth weight of patients in Group A and Group B was 3.00±0.61 and 3.12±0.75. The percentage of male patients was 57.1% and the percentage of female patients was 42.8%. The modified RDAI score in Group A and Group B at admission, 48 h of admission, and at the time of discharge was 4.34±0.87, 2.4±1.03, and 0.67±0.05 (p=0.04) and 4.11±0.58, 3.60±1.00, and 2.51±0.96 (p=0.12). The hospital stay was observed to be lowered in Group A (3.77±0.88) compared to Group B (5.43±0.92; p=0.04).&#x0D; Conclusion: From the findings of our study, we conclude that nebulized 3% hypertonic saline, as it acts by hindering the pathophysiologic mechanism of bronchiolitis, is more effective in reducing the clinical severity score and length of hospital stay. Further 3% hypertonic saline also have the additional benefit of decreasing the economic burden of disease as it is safe, inexpensive, reduces the inpatient hospital charges by reducing the length of stay.

Hyperosmolar salt solution in treatment of cerebral edema after severe craniocerebral injury

Clinical data of 93 patients with severe craniocerebral injury admitted in the Emergency Intensive Care Unit (EICU) of the First Affiliated Hospital of Zhengzhou University from September 2016 to September 2018 were retrospectively analyzed. Forty six patients received 10% hypertonic salt solution 60 ml (hypertonic salt group) and 47 patients received 20% mannitol 125 ml (mannitol group) for relieving early postoperation cerebral edema. The changes of intracranial pressure, central venous pressure, heart rate, mean arterial pressure (MAP), urine volume and serum sodium level at 2, 4 and 6 h after dehydrating agents were compared between two groups. There were no significant differences in the intracranial pressure, central venous pressure, heart rate and urine volume between two groups at 2, 4 and 6 h after the first dehydration treatment (all P>0.05). The MAP values of the two groups were (88±11) and (80±10), (85±10) and (78±9), (79±12) and (73±13) mmHg (1 mmHg=0.133 kPa) at 2, 4 and 6 h after the first dehydration treatment; and the serum sodium levels were (145±5) and (136±4), (144±6) and (133±5), (140±5) and (135±4) mmol/L, respectively. There were significant differences between two groups (all P<0.05). It is suggested that hypertonic salt can reduce intracranial pressure and increase cerebral perfusion better than mannitol in severe craniocerebral injury. Key words: Craniocerebral trauma; Intracranial hypertension; Saline solution, hypertonic; Comparative effectiveness research

DJ-1/PARK7 inhibits high glucose-induced oxidative stress to prevent retinal pericyte apoptosis via the PI3K/AKT/mTOR signaling pathway

Reactive oxygen species (ROS) act through multiple pathways to induce apoptosis of retinal capillary pericytes, which is an early marker and the primary cause of the progression of diabetic retinopathy. However, the specific molecular mechanisms behind ROS-induced retinal capillary pericyte loss in diabetic retinopathy remains elusive. In this study, we investigated the molecular regulation and effects of DJ-1/PARK7 on oxidative stress and injury of rat retinal pericytes (RRPs). To perform the research, RRPs were isolated from rat retina and cultured in medium with for 2 days: control group (5.6 mM glucose), high glucose group (30 mM glucose), hypertonic group (5.6 mM glucose + 24.4 mM mannitol). We found decreased expression of DJ-1 and increased apoptosis of RRPs in high glucose group. To further study the role of DJ-1, four groups were divided as follows: normal control group (5.6 mM glucose), high glucose (30 mM glucose), empty vector control group (pcDNA3.1,30 mM glucose), DJ-1 overexpression group (pcDNA3.1-myc-DJ-1,30 mM glucose). DJ-1, P53, p-P53, cleaved caspase-3, manganese superoxide dismutase (MnSOD), catalase (CAT) and PI3K/Akt/mTOR signaling pathway in each group was detected by Western Blot. RRPs apoptosis was detected by Terminal-deoxynucleoitidyl Transferase mediated Nick End Labeling (TUNEL) and 4′6- diamidino-2-phenylindole (DAPI). Mitochondrial function was detected by jc-1 and fluorescent probes DCFH-DA was used to determine reactive oxygen species (ROS). We found that high glucose (30 mM) lasting two days can induce significant apoptosis of RRPs, increase ROS production and expressions of p-p53 and active caspase-3, impair mitochondrial function, decrease the activities of MnSOD and CAT, and decrease expression of DJ-1, p-AKT and p-mTOR. In contrast, DJ-1/PARK7 overexpression significantly increases expression of DJ-1, p-AKT and p-mTOR, increases expression and activities of MnSOD and CAT, improves mitochondrial function, decreases expression of apoptotic gene protein p-p53 and active caspase-3, reduces ROS production and reduces the apoptotic rate of RRPs induced by high glucose. These results suggest that DJ-1 may play a role in protecting RRPs from high glucose induced-oxidative injury. DJ-1 might improve mitochondrial function, inhibit ROS production and enhance antioxidant capacity to reduce apoptosis of retinal pericytes through the PI3K/AKT/mTOR signaling pathway which may be related to early pathogenesis of diabetic retinopathy.

Bolus therapy with 3% hypertonic saline or 0.9% saline in emergency department patients with suspected sepsis: A pilot randomised controlled trial

Objective and designHypertonic saline administered during fluid resuscitation may mitigate endothelial glycocalyx (EG) shedding and inflammation. The objective of this pilot randomised controlled trial was to measure the effect of hypertonic saline, compared to isotonic saline, on biomarkers of EG shedding and inflammation in emergency department patients with suspected sepsis. MethodsPatients received either 5 mL/kg of 3% saline (hypertonic group, n = 34) or 10 mL/kg of 0.9% saline (isotonic group, n = 31). Change in serum biomarker concentrations of syndecan-1, hyaluronan, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, interleukin-6, -8, -10, interferon-γ, neutrophil gelatinase-associated lipocalin and resistin were compared from baseline (T0) to after fluid (T1), 3 h (T3) and 12–24 h (T24) later, as was serum osmolality, using linear mixed effects models. ResultsThe hypertonic group had significantly increased mean serum osmolality compared to the isotonic group at T1 (P < .001) and T3 (P = .004). Minor differences were found in some biomarker outcomes, including a decreased fold-change in syndecan-1 at T1 (P = .012) and in interleukin-10 at T24 (P = .006) in the isotonic group, compared to the hypertonic group. ConclusionsAlthough a single bolus of hypertonic saline increased serum osmolality, it did not reduce biomarkers of EG shedding or inflammation, compared to patients that received isotonic saline. Trial registrationANZCTR.org.au, ACTRN12611001021965, Registered on 23rd September 2011.

Spinal Hyper-Excitability and Altered Muscle Structure Contribute to Muscle Hypertonia in Newborns After Antenatal Hypoxia-Ischemia in a Rabbit Cerebral Palsy Model.

Rabbit kits after global antenatal hypoxic-ischemic injury exhibit motor deficits similar to humans with cerebral palsy. We tested several mechanisms previously implicated in spinal hyper-excitability after perinatal brain injury that may explain muscle hypertonia in newborns. Stiffness of hind limb muscles during passive stretch, electromyogram, and spinal excitability by Hoffman reflex, were assessed in rabbit kits with muscle hypertonia after global hypoxic-ischemic brain injury and naïve controls. Affected muscle architecture, motoneuron morphology, primary afferents density, gliosis, and KCC2 expression transporter in the spinal cord were also examined. Decrease knee stiffness after anesthetic administration was larger, but residual stiffness was higher in hypertonic kits compared to controls. Hypertonic kits exhibited muscle shortening and atrophy, in both agonists and antagonists. Sarcomere length was longer in tibialis anterior in hypertonic kits than in controls. Hypertonic kits had decreased rate dependent depression and increased Hmax/Mmax in H-reflex. Motor neuron soma sizes, primary afferent density were not different between controls and hypertonic kits. Length of dendritic tree and ramification index were lower in hypertonic group. Gene expression of KCC2 was lower in hypertonic kits, but protein content was not different between the groups. In conclusion, while we found evidence of decreased supraspinal inhibitory control and increased excitability by H-reflex that may contribute to neuronal component in hypertonia, increased joint resistance to stretch was explained predominantly by changes in passive properties of muscles and joints. We did not find structural evidence of increased sensory afferent input or morphological changes in motoneurons that might explain increased excitability. Gliosis, observed in spinal gray matter, may contribute to muscle hypertonia.

Effect of transforaminal hypertonic saline on the treatment of lumbar herniated discs: A prospective, randomized, controlled study

Aim: The aim of the study is to evaluate the effect of adding hypertonic saline to conventional transforaminal epidural steroid injections (TFESIs) in the management of patients with lumbar disc herniation (LDH) with radicular pain. Material and Methods: The study included 57 patients with L4–L5 LDH treated with TFESI. The patients were divided into two groups: the hypertonic (n = 29) and conventional groups (n = 28). The conventional group was administered with triamcinolone, bupivacaine, and isotonic saline. Subsequently, the hypertonic group was administered with triamcinolone, bupivacaine, and 10% hypertonic saline. The patients were followed up using the numeric rating scale (NRS) and Oswestry disability index (ODI). Substantial and moderate responder ratios were achieved using NRS. Outcome measurements were performed at baseline, one, three, six, and nine months. Complications were recorded in both groups. Results: TFESI with or without adjuvant hypertonic saline was found to be effective by achieving a significant reduction in NRS and ODI scores during the overall follow-up period. Reduction of NRS scores at six- and nine-month follow-up and ODI scores at nine-month follow-up in the hypertonic group were significantly higher than those in the conventional group. Moreover, substantial response in the hypertonic group was significantly higher than that in the conventional group. Both groups had no complications. Conclusion: The administration of adjuvant 10% hypertonic saline to transforaminal injections enhances the treatment efficacy by increasing pain reduction and improving the quality of life in the late follow-up period.

Vitamin D3 Ameliorates High Glucose-Induced Podocyte Apoptosis

Objective: Diabetic kidney disease (DKD) is a severe kidney disease characterized by podocyte apoptosis, injury, and accumulation of extracellular matrices that ultimately lead to end-stage renal disease. Vitamin D3 was reported to provide renal protection in DKD through the Vitamin D receptor (VDR), however there was limited data on whether it has a protective effect on podocytes of DKD induced by high glucose. Methods: The experimental subjects were conditionally immortalized mouse podocytes (MPC-5), which were divided into 4 groups: NG (normal glucose) group, HM (Hypertonic) group, HG (high glucose) group, VD3 (HG +1,25-dihydroxyvitamin D3 (1,25-D3)) group. Western-blot analysis was performed to detect the expression of Bcl-2, Bax and Caspase-3 in podocytes. The mRNA Expression of Bcl-2, Bax and Caspase-3 in the three groups were detected by RT-PCR method. Results: Compared with the NG group, the expression of apoptosis proteins Caspase-3 and Bax in the podocytes of HG group increased, and the expression of Bcl-2 decreased. Pretreatment with vitamin D3 attenuated these abnormalities in podocytes in HG group. Conclusion: Vitamin D can alleviate the apoptosis of podocytes induced by high glucose in vitro.

High salt intake increases plasma trimethylamine N-oxide (TMAO) concentration and produces gut dysbiosis in rats.

A high-salt diet is considered a cardiovascular risk factor; however, the mechanisms are not clear. Research suggests that gut bacteria-derived metabolites such as trimethylamine N-oxide (TMAO) are markers of cardiovascular diseases. We evaluated the effect of high salt intake on gut bacteria and their metabolites plasma level. Sprague Dawley rats ages 12-14 wk were maintained on either water (controls) or 0.9% or 2% sodium chloride (NaCl) water solution (isotonic and hypertonic groups, respectively) for 2 wk. Blood plasma, urine, and stool samples were analyzed for concentrations of trimethylamine (TMA; a TMAO precursor), TMAO, and indoxyl sulfate (indole metabolite). The gut-blood barrier permeability to TMA and TMA liver clearance were assessed at baseline and after TMA intracolonic challenge test. Gut bacterial flora was analyzed with a 16S ribosomal ribonucleic acid (rRNA) gene sequence analysis. The isotonic and hypertonic groups showed a significantly higher plasma TMAO and significantly lower 24-hr TMAO urine excretion than the controls. However, the TMA stool level was similar between the groups. There was no significant difference between the groups in gut-blood barrier permeability and TMA liver clearance. Plasma indoxyl concentration and 24-hr urine indoxyl excretion were similar between the groups. There was a significant difference between the groups in gut bacteria composition. High salt intake increases plasma TMAO concentration, which is associated with decreased TMAO urine excretion. Furthermore, high salt intake alters gut bacteria composition. These findings suggest that salt intake affects an interplay between gut bacteria and their host homeostasis.

The osmolyte type affects cartilage associated pathologic marker expression during in vitro mesenchymal stem cell chondrogenesis under hypertonic conditions.

Stem cells' fate during in vitro differentiation is influenced by biophysicochemical cues. Osmotic stress has proved to enhance chondrocyte marker expression, however its potent negative impacts had never been surveyed. We questioned whether specific osmotic conditions, regarding the osmolyte agent, could benefit chondrogenesis while dampening undesired concomitant hypertrophy and inflammatory responses. To examine the potential side effects of hypertonicity, we assessed cell proliferation as well as chondrogenic and hypertrophic marker expression of human Adipose Derived-MSC after a two week induction in chondrogenic media with either NaCl or Sorbitol, as the osmolyte agent to reach a +100 mOsm hypertonic condition. Calcium deposition and TNF-α secretion as markers associated with hypertrophy and inflammation were then assayed. While both hyperosmotic conditions upregulated chondrogenic markers, sorbitol had a nearly three times higher chondro-promotive effect and a lesser hypertrophic effect compared to NaCl. Also, a significantly lesser calcium deposition was observed in sorbitol hypertonic group. NaCl showed an anti-proinflammatory effect while sorbitol had no effect on inflammatory markers. The ossification potential and cartilage associated pathologic markers were affected differentially by the type of the osmolyte. Thus, a vigilant application of the osmotic agent is inevitable in order to avoid or reduce undesired hypertrophic and inflammatory phenotype acquisition by MSC during chondrogenic differentiation. Our findings are a step towards developing a more reliable chondrogenic regimen using external hypertonic cues for MSC chondrogenesis with potential applications in chondral lesions cell therapy.

Transient High-Glucose Stimulation Induces Persistent Inflammatory Factor Secretion from Rat Glomerular Mesangial Cells via an Epigenetic Mechanism

Background/Aims: Diabetic nephropathy is the one of the most serious microvascular complications of diabetes mellitus, and “metabolic memory” plays a vital role in the development of diabetic complications. To investigate the effect of epigenetics on metabolic memory, we analyzed the impact of transient high-glucose stimulation on the secretion of inflammatory factors from rat glomerular mesangial cells. Methods: Rat glomerular mesangial cells (HBZY-1) were divided into three groups: high-glucose group (25 mM glucose), hypertonic group (5.5 mM glucose+19.5 mM mannitol), and normal-glucose control group (5.5 mM glucose). Mesangial cells were cultured in high-glucose, hypertonic, and normal-glucose media for 24 h and transitioned to normal-glucose culture for 24, 48, and 72 h. Then, protein, mRNA, and supernatants were harvested. The expression of monomethylated H3K4 was determined by western blot analysis, and the expression of the NF-κB subunit p65 and histone methyltransferase set7/9 was determined by quantitative real-time PCR. The expression of monocyte chemoattractant protein 1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) was detected by an enzyme-linked immunosorbent assay. Results: Compared with the control group, H3K4me1 expression was upregulated after transient high-glucose stimulation, gradually downregulated in the following 48 h (P < 0.05), and reached the level of the control group at 72 h (P > 0.05). The expression of set7/9 was increased after 24 h of high-glucose stimulation and the following 24 h and 48 h (P < 0.05); it then returned to the level of the control group at 72 h. Compared with the control group, the increased expression of p65, VCAM-1, and MCP-1 was sustained for at least 72 h in the high-glucose group. Conclusion: Transient high-glucose stimulation can induce the persistent secretion of inflammatory factors from rat glomerular mesangial cells via histone modification.

Comparison of fluid balance and hemodynamic and metabolic effects of sodium lactate versus sodium bicarbonate versus 0.9% NaCl in porcine endotoxic shock: a randomized, open-label, controlled study

BackgroundSodium lactate has been shown to improve hemodynamics and avoid fluid overload. The objective of this study was to confirm a beneficial effect on fluid balance with sodium lactate infusion and to specify whether the advantage of lactate is related to a negative chloride balance, its particular metabolism, or simply its energy load.MethodsThis was an interventional, randomized, open-label, controlled experimental study. Fifteen female “large white” pigs (2 months old) were challenged with intravenous infusion of Escherichia coli endotoxin. Three groups of five animals were randomly assigned to receive different fluids: a treatment group received sodium lactate 11.2% (SL group); an isotonic control group received 0.9% NaCl (NC group); and a hypertonic control group, with the same amount of osmoles and sodium as the SL group, received sodium bicarbonate 8.4% (SB group). In order to provide the same energy load in the three groups, control groups were perfused with an equivalent energy supply. Statistical analysis was performed with non-parametric tests and the Dunn correction for multiple comparisons at p < 0.05.ResultsFluid and chloride balance, hemodynamics, oxygenation markers, and microcirculatory parameters were measured over a 5-h period. Cumulative fluid balance was significantly lower in the SL group (550 (415–800) mL; median (interquartile range)) compared to the NC group (1100 (920–1640) mL, p = 0.01) and the SB group (935 (790–1220) mL, p = 0.03). Hemodynamics, cardiac efficiency, and microcirculation were significantly enhanced in the SL group, resulting in a significant improvement in oxygen delivery (SL group 417 (305–565) mL/min/m2 at 300 min versus the NC (207 (119–272) mL/min/m2, p = 0.01) and the SB (278, (211–315) mL/min/m2, p = 0.03) groups). Oxygenation markers (arterial oxygen partial pressure (PaO2)/inspired oxygen fraction (FiO2), mixed venous oxygen saturation (SvO2), and venoarterial carbon dioxide tension difference (Pv-aCO2) were enhanced with sodium lactate infusion. Chloride balance was equivalent in both hypertonic groups and significantly reduced compared to the NC group.ConclusionSodium lactate infusion improves fluid balance and hemodynamics. The advantage of lactate does not seem to be explained by its energy load or by the induced negative chloride balance with subsequent water movements.

Hyperoxia and hypertonic saline in patients with septic shock (HYPERS2S): a two-by-two factorial, multicentre, randomised, clinical trial

There is insufficient research into the use of mechanical ventilation with increased inspiratory oxygen concentration (FiO2) and fluid resuscitation with hypertonic saline solution in patients with septic shock. We tested whether these interventions are associated with reduced mortality. This two-by-two factorial, multicentre, randomised, clinical trial (HYPERS2S) recruited patients aged 18 years and older with septic shock who were on mechanical ventilation from 22 centres in France. Patients were randomly assigned 1:1:1:1 to four groups by a computer generated randomisation list stratified by site and presence or absence of acute respiratory distress syndrome by use of permuted blocks of random sizes. Patients received, in an open-labelled manner, mechanical ventilation either with FiO2 at 1·0 (hyperoxia) or FiO2 set to target an arterial haemoglobin oxygen saturation of 88-95% (normoxia) during the first 24 h; patients also received, in a double-blind manner, either 280 mL boluses of 3·0% (hypertonic) saline or 0·9% (isotonic) saline for fluid resuscitation during the first 72 h. The primary endpoint was mortality at day 28 after randomisation in the intention-to-treat population. This study was registered with ClinicalTrials.gov, number NCT01722422. Between Nov 3, 2012, and June 13, 2014, 442 patients were recruited and assigned to a treatment group (normoxia [n=223] or hyperoxia [n=219]; isotonic [n=224] or hypertonic [n=218]). The trial was stopped prematurely for safety reasons. 28 day mortality was recorded for 434 patients; 93 (43%) of 217 patients had died in the hyperoxia group versus 77 (35%) of 217 patients in the normoxia group (hazard ratio [HR] 1·27, 95% CI 0·94-1·72; p=0·12). 89 (42%) of 214 patients had died in the hypertonic group versus 81 (37%) of 220 patients in the isotonic group (HR 1·19, 0·88-1·61; p=0·25). We found a significant difference in the overall incidence of serious adverse events between the hyperoxia (185 [85%]) and normoxia groups (165 [76%]; p=0·02), with a clinically relevant doubling in the hyperoxia group of the number of patients with intensive care unit-acquired weakness (24 [11%] vs 13 [6%]; p=0·06) and atelectasis (26 [12%] vs 13 [6%]; p=0·04) compared with the normoxia group. We found no statistical difference for serious adverse events between the two saline groups (p=0·23). In patients with septic shock, setting FiO2 to 1·0 to induce arterial hyperoxia might increase the risk of mortality. Hypertonic (3%) saline did not improve survival. The French Ministry of Health.

Efficacy of Adjuvant 10% Hypertonic Saline in Transforaminal Epidural Steroid Injection: A Retrospective Analysis

Chronic lower back pain with or without radiculopathy represents an important medical, social, and economic problem. Many treatment modalities and techniques, including surgery and epidural administration of steroids, have been used to manage this pain. Hypertonic saline, which has been used as an adjunct to percutaneous epidural adhesiolysis, can also be injected via a transforaminal approach in expectation of longer-lasting effects. This study aimed to determine the effect of adding hypertonic saline to conventional transforaminal epidural steroid injections (TFEI) to provide pain relief for chronic radiculopathy patients. A retrospective study. Pain clinic of a university hospital. Between January 2010 and December 2013, the medical records of 246 patients (94 in the hypertonic group, 153 in the control group) who received transforaminal epidural block were reviewed and analyzed. The hypertonic group received 10% sodium chloride solution added to lidocaine, triamcinolone, and hyaluronidase. Outcomes on pain reduction were measured using a numerical rating scale (NRS) and the responder rate at baseline, one, 3, and 6 months after procedure. The estimated difference in NRS scores from baseline throughout a 6-month follow-up period in the hypertonic group were significantly higher (P = 0.0003). The proportion of substantial responders (41.9% vs. 34.6% at one month, 40.9% vs. 26.8% at 3 months, and 33.3% vs. 14.4% at 6 months, respectively, P = 0.0058) and substantial/moderate responders (71.0% vs. 58.8% at one month, 65.6% vs. 40.4% at 3 months, and 48.4% vs. 20.3% at 6 months, respectively, P < 0.0001) were significantly higher in the hypertonic group. The Oswestry disability index (ODI) was not different between the groups (P = 0.2697). Retrospective design without a control group. Hypertonic saline provides more superior and longer lasting pain relieving effects when added to TFEIs.Key words: Back pain, epidural injections, epidural steroids, hypertonic saline, lumbar, radiculopathy, transforaminal.

Effect of ALDH2 on High Glucose-Induced Cardiac Fibroblast Oxidative Stress, Apoptosis, and Fibrosis.

Our study aimed firstly to observe whether ALDH2 was expressed in neonate rat cardiac fibroblasts, then to investigate the effect of activation of ALDH2 on oxidative stress, apoptosis, and fibrosis when cardiac fibroblasts were subjected to high glucose intervention. Cultured cardiac fibroblasts were randomly divided into normal (NG), NG + Alda-1, high glucose (HG), HG + Alda-1, HG + Alda-1 + daidzin, HG + daidzin, and hypertonic groups. Double-label immunofluorescence staining, RT-PCR, and Western blot revealed ALDH2 was expressed in cardiac fibroblasts. Compared with NG, ALDH2 activity and protein expression were reduced, and cardiac fibroblast proliferation, ROS releasing, 4-HNE protein expression, collagen type I and III at mRNA levels, and the apoptosis rate were increased in HG group. While in HG + Alda-1 group, with the increases of ALDH2 activity and protein expression, the cardiac fibroblast proliferation and ROS releasing were decreased, and 4-HNE protein expression, collagen type I and III at mRNA levels, and apoptosis rate were reduced compared with HG group. When treated with daidzin in HG + Alda-1 group, the protective effects were inhibited. Our findings suggested that ALDH2 is expressed in neonate rat cardiac fibroblasts; activation of ALDH2 decreases the HG-induced apoptosis and fibrosis through inhibition of oxidative stress.