The objective of this study was to explore the possible effects of leukocyte elimination by filteration before storage on the quality of red blood cell concentrations (RCC) that prepared through two procedures. Eight units of red blood cell concentrations derived from whole blood after plasma separated (RCC1) and eight units of red blood cell concentrations derived from whole blood after platelet-rich plasma separated (RCC2) were divided randomly into filtered group and control group respectively. The RCC of filtered group were filtered by leukocyte deplete filter before storage. The control group didn't have any other treatments. These two groups were stored for five weeks at 4 degrees C according to AABB standard. Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and plasma concentration of K(+) and lactate dehydrogenase (LDH), free hemoglobin (FHb), adenosine triphosphate (ATP) of red blood cell of all RCC were evaluated weekly, and bacteria contamination of all RCC was also detected after five weeks of storage. The results showed that there was no difference of MCV, MCH and MCHC and ATP level of red blood cell in all RCC of two groups, the ATP of red blood cell was lower than the control group on week 4 and 5. The average concentration of K(+) of the filtered group was less than the control group. The differences are significant except that of RCC1 stored till the third week. The plasma LDH concentration of filtered group was less than the control group, and the differences were exacerbate during the storing time prolonged. FHb release in the filtered group of RCC2 was significant less than that of control, but no significant difference was found between the two groups of RCC1. It was concluded that leukocyte elimination by filter before storage could be benefit to RCC preservation.
Adenosine Triphosphate ; metabolism ; Blood Component Removal ; Blood Preservation ; Erythrocytes ; physiology ; Filtration ; Humans ; L-Lactate Dehydrogenase ; blood ; Leukocytes ; Potassium ; blood

In order to investigate the effect of antioxidants on human blood, vitamin C was selected and added into plastic blood storage bags with CPD, and stored at 25 degrees C. During 6 days of storage, some indexes as ATP, SOD, MDA, K(+) concentration and superoxide radicals were detected and were compared with control group, The results showed that ATP and SOD activity in whole blood with vitamin C during 6 days of storage was higher then that in control group (P < 0.05, P < 0.01), the MDA and plasma K(+) concentrations in stored whole blood with vitamin C during 6 days of storage were lower than that in control group (P < 0.05, P < 0.01), the superoxide radical concentrations in stored whole blood with vitamin C decreased lower than that in control group (30%). The conclusion was made that vitamin C increases activities of ATP and SOD, decreases concentrations of MDA, plasma K(+) and superoxide radicals during blood preservation.
Adenosine Triphosphate ; blood ; Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Blood Preservation ; methods ; Erythrocytes ; cytology ; drug effects ; metabolism ; Humans ; Malondialdehyde ; blood ; Potassium ; blood ; Superoxide Dismutase ; blood ; Time Factors

Chronic stable diabetic patients (n = 6) were compared with healthy control subjects (n = 5) after acute oral intake of 50 mEq of potassium chloride (KCl) to investigate for possible derangements of homeostatic responses for acute term (3 hrs) to acute potassium load. Plasma renin activity (PRA), plasma aldosterone (PA), and transtubular potassium concentration gradient (TTKG) known as a useful semiquantative index of distal nephron potassium secretion were measured. All the baseline parameters were comparable between diabetic and non-diabetic subjects except for significantly reduced creatinine clearance in diabetics (mean +/- SEM, 105 +/- 4 vs. 85 +/- 5 ml/min, p < 0.05). Following acute oral KCl load, the peak increases of serum potassium changes from basal levels were noted at 2 hours in both groups, but were higher in diabetic subjects (mean +/- SEM, 0.42 +/- 0.06 vs. 0.62 +/- 0.09 mEq/L). Also, 4 out of 6 diabetic subjects but none of the control subjects at 2 hours after oral KCl load became hyperkalemic ( > 5.0 mEq/L). PRA did not show any significant changes, whereas PA was increased simultaneously with increments in serum potassium in both groups, with blunted increases in the diabetics. However, TTKG was increased prominently in control subjects (8.18 from 4.98), but only slightly in diabetic subjects (4.55 from 4.18), with statistical difference between the two groups (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Adult ; Aged ; Aldosterone/blood ; Diabetes Mellitus, Type 2/*metabolism ; *Homeostasis ; Humans ; Kidney Tubules/metabolism ; Male ; Middle Aged ; Potassium/*metabolism ; Renin/blood

OBJECTIVETo investigate the postburn changes in gastric mucosal blood flow (GMBF) and Na(+)-K(+)-ATPase activity and their effects on gastric transmucosal potential difference (GTPD) in severely scalded rats.

METHODSRats were inflicted with hot water scald on the back. Laser Doppler blood flow meter, electric physiological recorder meter and biochemical method were employed in the study to determine the changes in GMBF, GTPD and gastric mucosal Na(+)-K(+)-ATPase activity in the rats before injury and at 3, 6, 12, 24 and 48 PBHs. Normal rats served as controls, The above indices were also detected at the corresponding time points. All the data was collected and analyzed for their correlation.

RESULTSThe GMBF and Na(+)-K(+)-ATPase activity was at 3 - 24 PBHs were evidently lower in the scalded rats than those in controls (P < 0.05 - 0.01). GTPD was decreased significantly at 6 - 48 PBHs (P < 0.05 - 0.01). was above three indices reached the lowest values at 12 PBH. By correlation analysis the results indicated that the Na(+)-K(+)-ATPase activity The decreased in accordance with a decrease in GMBF (r = 0.527, P < 0.01). The GTPD decrease could both be induced by GMBF decrease and Na(+)-K(+)-ATPase activity decrease (r = 0.453 and 0.527, respectively, P < 0.01).

CONCLUSIONThe decrease in GMBF and Na(+)-K(+)-ATPase activity occurred in severely scalded rats, both changes might be the major cause of postburn gastric mucosal barrier injury.

Animals ; Burns ; metabolism ; physiopathology ; Gastric Mucosa ; blood supply ; metabolism ; physiopathology ; Male ; Membrane Potentials ; Rats ; Rats, Wistar ; Sodium-Potassium-Exchanging ATPase ; metabolism

Hypertension is one of the strongest risk factors for cardiovascular diseases, cerebral stroke, and kidney failure. Lifestyle and nutrition are important factors that modulate blood pressure. Hypertension can be controlled by increasing physical activity, decreasing alcohol and sodium intake, and stopping tobacco smoking. Chronic kidney disease patients often have increased blood pressure, which indicates that kidney is one of the major organs responsible for blood pressure homeostasis. The decrease of renal sodium reabsorption and increase of diuresis induced by high potassium intake is critical for the blood pressure reduction. The beneficial effect of a high potassium diet on hypertension could be explained by decreased salt reabsorption by sodium-chloride cotransporter (NCC) in the distal convoluted tubule (DCT). In DCT cells, NCC activity is controlled by with-no-lysine kinases (WNKs) and its down-stream target kinases, Ste20-related proline-alanine-rich kinase (SPAK) and oxidative stress-responsive 1 (OSR1). The kinase activity of WNKs is inhibited by intracellular chloride ([Cl^-]_i) and WNK4 is known to be the major WNK positively regulating NCC. Based on our previous studies, high potassium intake reduces the basolateral potassium conductance, decreases the negativity of DCT basolateral membrane (depolarization), and increases [Cl^-]_i. High [Cl^-]_i inhibits WNK4-SPAK/OSR1 pathway, and thereby decreases NCC phosphorylation. In this review, we discuss the role of DCT in the blood pressure regulation by dietary potassium intake, which is the mechanism that has been best dissected so far.
Blood Pressure ; Diet ; Humans ; Kidney/metabolism* ; Kidney Tubules, Distal/metabolism* ; Phosphorylation ; Potassium/pharmacology* ; Protein Serine-Threonine Kinases ; Solute Carrier Family 12, Member 3/metabolism*

BACKGROUNDPotassium (K+) channels are important in regulating cell membrane potential and excitability. Although bronchial myocytes from asthmatic rats show a significant reduction in voltage-dependent delayed rectifier potassium channel (Kv) current density and higher excitability, the activity and expression of Kv in human bronchial smooth muscle cells (HBSMCs) have never been studied. The objective of this study was to investigate the effect of passive sensitization by asthmatic serum on the activity of Kv and the expression of Kv isoform Kv1.5 in HBSMCs.

METHODSHBSMCs were randomly divided into two groups: control group (containing 10% serum from nonatopic individuals) and sensitized group (containing 10% asthmatic serum), then cultured for 24 hours. Whole-cell patch clamp, immunofluorescence staining, reverse transcription-polymerase chain reaction and Western blot techniques were used to study the effect of passive sensitization on the activity of Kv and the expression of Kv1.5 in HBSMCs.

RESULTSThe membrane potential in passively sensitized HBSMCs was significantly depolarized to -(26.7 +/- 5.2) mV compared with -(41.3 +/- 6.4) mV in the control group (P < 0.01). Passive sensitization caused a significant inhibition of Kv currents in HBSMCs, resulting in a downward shift in the current-voltage (I-V) relationship curve. At +50 mV, the peak Kv current density of passively sensitized HBSMCs was significantly decreased from (54.6 +/- 8.7) picoamperes per picofarad (pA/pF) to (32.1 +/- 7.1) pA/pF (P < 0.01). The expression level of Kv1.5 mRNA in passively sensitized HBSMCs was significantly lower than that in the control group (0.76 +/- 0.07 vs 1.04 +/- 0.13, P < 0.05). The expression of Kv1.5 protein of passively sensitized HBSMCs was also significantly reduced compared to that from the control group (984 +/- 168 vs 2200 +/- 380, P < 0.05).

CONCLUSIONSThe activity and expression of Kv were all decreased in HBSMCs passively sensitized by asthmatic serum compared with nonsensitized cells. These changes might be involved in the mechanisms of formation and development of asthma.

Asthma ; blood ; Bronchi ; metabolism ; Cells, Cultured ; Female ; Fluorescent Antibody Technique ; Humans ; Immunization ; Kv1.5 Potassium Channel ; Male ; Myocytes, Smooth Muscle ; metabolism ; Potassium Channels, Voltage-Gated ; analysis ; genetics ; RNA, Messenger ; analysis

OBJECTIVETo study the expression and mechanism of WNK4 gene regulated by aldosterone.

METHODSWistar rats were treated with aldosterone and potassium water. Serum aldosterone and ion as well as urine ion were measured. The expression of WNK4 gene in kidney tissues was detected by real-time PCR. Kidney-derived HEK293 cells were cultured, transfected with pGL3-WNK4, and then stimulated by aldosterone. After 24 h of transfection, luciferase activities of the plasmid were detected.

RESULTSCompared with those of the controls, serum aldosterone and urine K(+) of experimental rats were significantly elevated, whilst urine Na(+) was significantly decreased. And urine Cl(-) was significantly increased only in the group of high K(+). Serum K(+), Na(+) and Cl(-) showed no significant difference. Expression of WNK4 gene in kidney tissues was significantly decreased. The luciferase activity of pGL3-WNK4-484 plasmid has decreased after stimulated with aldosterone, while the activity of pGL3-WNK4-275 showed no change.

CONCLUSIONAldosterone can down-regulate the expression of WNK4 through binding with regulatory element in the upstream of the gene.

Aldosterone ; blood ; pharmacology ; Animals ; Cell Line ; Chlorides ; blood ; Gene Expression Regulation ; drug effects ; Humans ; Kidney ; metabolism ; Male ; Potassium ; blood ; Promoter Regions, Genetic ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Rats ; Sodium ; blood ; Transcriptional Activation ; drug effects

AIMTo investigate the changes of large-conductance calcium-activated potassium channels (BKCa, MaxiK) during aging and relations between the changes and blood pressure.

METHODSMale spontaneously hypertensive rats (SHR) aged 9, 15, 21, 27, 33 weeks (the number of each weeks SHR was 4) were selected as hypertension group rats, corresponding gender, weeks and number Wistar-Kyoto rats (WKY) as control group rats. Blood pressure of abdominalis aorta of each weeks SHR and WKY were measured by BL-420F experimental system of biological function. The arteria mesenteric minor (AMM) were isolated in blunt dissection method. The vascular smooth muscle cells (VSMCs) of AMM were isolated with prolease. The potassium current, the current after BKCa were blockaded by Tetraethylammonium (TEA) and the capacitance of membrane (Cm) of VSMCs of AMM were recorded with using whole cell patch clamp, and calculated the BKCa current and the BKCa current density. Probe the correlation of the changes of BKCa current density with MABP during aging.

RESULTSThe potassium current density and BKCa current density of VSMCs of AMM of SHR were decreasing during aging, however, the changes of WKY had no statistically significance (P > 0.05). The BKCa current density was extremely correlative with MABP in SH R (the values of r were -0.7174), in WKY, the BKCa current density was correlative with MAB P r = -0.4832.

CONCLUSIONBKCa current and current density attenuate with aging, the level of blood pressure is response of the attenuated degree. The BKCa current density is extremely correlative with the blood pressure.

Aging ; physiology ; Animals ; Blood Pressure ; physiology ; Cell Membrane ; physiology ; Hypertension ; metabolism ; physiopathology ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; physiology ; Male ; Membrane Potentials ; physiology ; Mesenteric Arteries ; cytology ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; physiology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

AIMTo observe the neuroprotective effect of limb ischemic post-conditioning (LIPC) on local brain ischemia and reperfusion injury in rat, and to investigate whether mitochondrial ATP sensitive potassium channel (mito K(ATP)) and kappa-opioid receptor were involved in the neuroprotection.

METHODSRats were randomly divided into 6 groups that were ischemia/reperfusion group, unilateral hindlimb ischemia group (uLIPC), bilateral hindlimbs ischemia group (bLIPC), bLIPC + antagonist of kappa-opioid receptor nor-binaltorphimine (nor-BNI) group, bLIPC + mito K(ATP) blocker 5-hydroxydecanoate(5-HD) group, bLIPC + extracorporeal circulation of bilateral hindlimbs via femoral arteries (EC) group. Cerebral ischemia was induced by middle cerebral artery occlusion (MCAO), neurological scores, plasma levels of dynorphin and enkephalin, the brain infarct areas were determined after reperfusion.

RESULTSUnilateral LIPC partially improved the neurological score after local brain ischemia and reperfusion injury in rat (P < 0.05), and decreased the infarct area compared with the untreated group undergoing brain ischemia and reperfusion (P < 0.01). Bilateral LIPC significantly improved the neurological score after local brain ischemia and reperfusion injury (P < 0.01), and decreased the infarct area (P < 0.01). The neurological scores of bilateral LIPC group were significant higher than those of unilateral LIPC (P < 0.05). The plasma level of dynorphin was significantly increased (P < 0.01) at 5, 15, 30 min, 1 and 2 h after bilateral LIPC, however, it deceased to the normal level at 12 h after bilateral LIPC. The plasma level of enkephalin showed no obvious change after bilateral LIPC (P > 0.05). nor-BNI (25 nmol/L) and 5-HD (10 mg/kg) abolished the effect of bilateral LIPC (P < 0.01).

CONCLUSIONLIPC protects rat from local brain ischemia and reperfusion injury. Mito K(ATP) may be involved in the neuroprotection, and kappa-opioid receptor may also participate in the protective effect.

Animals ; Brain Ischemia ; physiopathology ; Extremities ; blood supply ; Ischemic Postconditioning ; methods ; Male ; Potassium Channels ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, kappa ; metabolism ; Reperfusion Injury ; prevention & control

OBJECTIVEStudies have shown that potassium channel plays a pivotal role in T cell activation. The expression of potassium channel gene KCTD9 was evidenced being highly upregulated in patients with severe hepatitis B (SHB). To understand this phenomenon further, tissue and cellular expression profiles of KCTD9 were investigated in patients with SHB.

METHODSA rabbit peptide polyclonal antibody was prepared. Various samples including peripheral blood mononuclear cells (PBMCs); livers from patients with SHB or mild chronic hepatitis B, were examined for KCTD9 expression by quantitative real time PCR and immunohistochemistry staining (IHC). Confocal microscopy was used to illustrate the localizations of the expressions.

RESULTSIncreased expression of KCTD9 was observed in PBMC in over 35.7% of the patients with SHB when compared with that of patients with mild chronic hepatitis B. In all patients, the relative value of increased KCTD9 mRNA was positively correlated with alanine aminotransferase, aspartate aminotransferase, total bilirubin and direct bilirubin but negatively with serum albumin. The expression was mainly located in hepatocytes, bile duct epithelial cells, Kupffer cells and inflammatory cells, and in the cytoplasm of PBMCs from the healthy individuals and patients with mild chronic hepatitis B, whereas in both cytoplasm and nuclei in those from patients with SHB.

CONCLUSIONThe increased expression of potassium channel gene KCTD9 correlates with disease severity in patients with viral hepatitis B.

Adult ; Female ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Middle Aged ; Monocytes ; metabolism ; Potassium Channels ; genetics ; metabolism ; RNA, Messenger ; genetics ; Young Adult