BACKGROUND:Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cels (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs. OBJECTIVE:To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplificationin vitro. METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cel surface-associated antigens; cel counting kit-8 was used to detect cel proliferation; RT-PCR was used to determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cel-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α. RESULTS AND CONCLUSION:The cels were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed thatunder hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cel-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionaly, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200μmol/L. However, a higher concentration of CoCl2 (≥ 250μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.

Objective To investigate the effect of atorvastatin on the expressions of long non-coding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1)and inflammation factors in human umbilical vein endothelial cells (HUVECs) stimulated by high glucose. Methods The expression of MALAT1 in HUVECs incubated with high glucose(30 mmol/L) for different time periods were detected by real-time PCR. Under high glucose condition, the expressions of MALAT1, interleukin-6(IL-6), and interleukin-8 (IL-8) in HUVECs were detected after MALAT1 was silenced by siRNA or atorvastatin was added. Results (1) After HUVECs were incubated with high glucose for different time periods, the expressions of MALAT1 were increased to some extent(P<0.05), with the peak at 12h (P<0.01). The levels of IL-6 and IL-8 expression and secretion were increased after HUVECs were stimulated by high glucose for 12h (P<0.05). (2)The silence of MALAT1 markedly suppressed high glucose-stimulated expression and secretion of IL-6 and IL-8 (P<0.05). (3) Atorvastatin significantly inhibited high glucose-stimulated expressions of MALAT1, IL-6, and IL-8(all P<0.05). Conclusions High glucose induces the secretion of inflammatory factors by stimulating MALAT1 expression in endothelial cells. Atorvastatin significantly inhibits high glucose-stimulated MALAT1 expression and decreases inflammatory reaction.

BACKGROUNDVisfatin, a visceral fat-derived adipocytokine, plays a significant physiological function in lipid metabolism. However, the precise function of visfatin and its regulation by thyroid hormones are still unknown. This study observed the plasma visfatin concentrations in subjects with hyperthyroidism and hypothyroidism in vivo, and investigated the possible regulation mechanism between visfatin and tri-iodothyronine (T3) in vitro as a further interpretation.

METHODSThe experiment in vivo included clinical subjects (57 patients with thyroid dysfunction and 29 euthyroid healthy volunteers) and an animal model (24 Wistar rats). All subjects were divided into hyperthyroidism, hypothyroidism and euthyroidism groups, with plasma thyroid hormones, thyrotropin, visfatin and triglyceride concentrations determined. Visfatin mRNA expression in visceral fat and liver of rats was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). The experiment in vitro studied 3T3-L1 cells and visfatin mRNA expression under nine different T3 concentrations (0, 0.1, 0.25, 0.5, 1, 5, 10, 20, 100 nmol/L) using quantitative real-time RT-PCR.

RESULTSClinical subjects and animal models showed elevated plasma visfatin concentrations in the hyperthyroidism group (20.466 ng/ml (15.263, 26.795 ng/ml) and (1209.164±165.292) ng/L) and hypothyroidism group (12.457 ng/ml (11.115, 15.454 ng/ml) and (1205.425±109.200) ng/L) compared to euthyroidism group (6.891 ng/ml (5.888, 8.803 ng/ml) and (926.650±54.002) ng/L, P<0.001). For animal models, visfatin mRNA expression in visceral fat in the hyperthyroidism and hypothyroidism groups increased about 3.33-fold and 1.98-fold compared to the euthyroidism group (P<0.001), which was positively correlated with plasma visfatin concentrations (r=0.713, P<0.001). However, no significant group difference (P>0.05) and correlation (r=0.121, P=0.572) was found in the liver. T3 induced a remarkable increase of visfatin mRNA expression in 3T3-L1 cells at low concentrations (0-0.5 nmol/L T3) followed by a sharp decrease at higher concentrations (0.5-100 nmol/L T3), with an inflection point at 0.5 nmol/L T3.

CONCLUSIONElevated circulating visfatin levels in subjects with hyperthyroidism and hypothyroidism are possibly due to an increase of visfatin mRNA expression in visceral fat, and a nonlinear regulation mechanism on visfatin mRNA expression under various T3 concentrations might be involved.

3T3-L1 Cells ; Adult ; Animals ; Female ; Humans ; Hyperthyroidism ; blood ; genetics ; metabolism ; Hypothyroidism ; blood ; genetics ; metabolism ; Male ; Mice ; Middle Aged ; Nicotinamide Phosphoribosyltransferase ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Triiodothyronine ; blood

OBJECTIVETo study the changes of serum albumin contents after operation and investigate whether post-operational serum albumin contents are correlated with the disease severity in children with acute intussusception.

METHODSSerum albumin contents were measured using the automatic biochemistry analyzer in 32 children with mild acute intussusception and 21 children with severe acute intussusception 1 day after surgical operation. After 5 days combined treatment, serum albumin contents were re-examined. Thirty healthy children severed as the control group. The correlation between post-operational serum albumin contents and critical illness scores was evaluated.

RESULTSSerum albumin contents in the mild (34.2+/-6.5 g/L; P<0.05) and the severe intussusception groups (25.8+/-7.5 g/L; P<0.01) 1 day after operation were significantly lower than those in the control group (37.1+/-4.1 g/L). There were significant differences in serum albumin contents between the mild and the severe intussusception groups (P<0.05). Five days after operation, serum albumin contents in the mild intussusception group significantly increased (37.1+/-11.4 g/L; P<0.05), while serum albumin contents in the severe intussusception group did not differ from those 1 day after operation. There was a positive correlation between serum albumin contents on the 1st day after operation and the critical illness scores (r=0.879, P<0.01).

CONCLUSIONSSerum albumin contents decreased on the 1st day after operation and were correlated with the disease severity in children with acute intussusception. Hypoalbuminemia lasted for a longer period in severe cases. The post-operational measurement of serum albumin contents may be useful in the evaluation of the severity for children with acute intussusception.

Acute Disease ; Child, Preschool ; Female ; Humans ; Infant ; Intussusception ; blood ; complications ; surgery ; Male ; Serum Albumin ; analysis

ObjectiveTo investigate the expression and significance of hypoxia-inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) in liver tissues after ischemia-reperfusion injury (IRI) in rats.MethodsSixteen Sprague-Dawley (SD) rats were randomized into the IRI group and the Sham group according to the random number table, 8 rats in each group. The portal vein and branches of hepatic artery of the rats in the IRI group were clamped with the atraumatic vascular clamp. After 45 min of ischemia, reperfusion was performed for 6 h. The porta hepatis of rats in the Sham group was exposed and the hepatic blood flow was not occluded. The morphological changes of the liver tissues in two groups were observed. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were tested and the contents of HIF-1α and VEGF in liver tissues were determined. Data comparison between two groups was conducted usingt test.ResultsUnder light microscope, swelling, vacuolar degeneration of the liver cells, inflammatory cell infiltration, hepatic sinusoidal dilatation, disorganized hepatic cords and spotty necrosis were observed in the IRI group. The histomorphology of liver tissues in the Sham group was normal. The serum ALT and AST in the IRI group was respectively (1 007±130) and (1 307±72) U/L, which were significantly higher than (59±4) and (81±3) U/L in the Sham group (t=20.700, 48.448;P<0.05). The relative expression of HIF-1α and VEGF in liver tissues of IRI group was 1.77±0.10 and 1.86±0.05 respectively, which were significantly higher than 0.60±0.22 and 0.83±0.06 in the Sham group (t=13.623, 35.563;P<0.05).ConclusionThe expression of HIF-1αand VEGF in liver tissues increase significantly during early IRI, which may play a role in reducing the IRI of organism.

OBJECTIVETo explore the mechanisms of Chinese herbal medicine Sanqi Oral Liquid, composed of Astragalus membranaceus and Panpax notoginseng, in alleviating renal injury by observing its effect on the expressions of CD4(+), CD8(+) and CD68(+) cells in 5/6 nephrectomized rats with chronic renal failure.

METHODSA total of 102 SD rats were randomly divided into six groups: three treatment groups were administrated with high, medium and low dosage of Sanqi Oral Liquid respectively by gavage; a normal group, a 5/6 nephrectomized model group, and a group treated with coated aldehyde oxygenstarch were used as controls. Following oral administration of Sanqi Oral Liquid for 12 weeks, the general condition and renal pathological changes were observed, and the renal function, platelet count (PLT) and the expressions of CD4(+), CD8(+) and CD68(+) cells were determined for each group.

RESULTSThere were proliferation of mesangial matrix, renaltubularnecrosis and obvious tubulointerstitial fibrosis in the model group, and they were much milder in the treatment groups. Compared with the model group, the amounts of blood urea nitrogen (BUN), serum creatinine (Scr) and PLT in the treatment groups decreased (P<0.05 for all); and in the group administrated of medium dosage of Sanqi Oral Liquid, the expression of CD4(+) cells was up-regulated and those of CD8(+) and CD68(+) cells were down-regulated (P<0.05 for all), leading to an increased ratio of CD4(+)/CD8(+)(P<0.01).

CONCLUSIONSanqi Oral Liquid has a significant effect on regulating lymphocyte subsets, reducing the infiltration of macrophages in renal tissues and alleviating tubulointerstitial fibrosis, and this may be one of mechanisms of Sanqi Oral Liquid in delaying the progression of chronic kidney diseases.

Administration, Oral ; Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Astragalus membranaceus ; chemistry ; CD4-Positive T-Lymphocytes ; drug effects ; pathology ; physiology ; CD8-Positive T-Lymphocytes ; drug effects ; pathology ; physiology ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Kidney Failure, Chronic ; drug therapy ; immunology ; pathology ; surgery ; Lymphocyte Count ; Male ; Nephrectomy ; Panax notoginseng ; chemistry ; Rats ; Rats, Sprague-Dawley ; Solutions

OBJECTIVETo evaluate the process of smoking prevention and control among Chinese adolescent with a model on health promotion in junior high school.

METHODSA cluster-randomized, controlled and schooled-based trial on smoking prevention was conducted among 2343 students at four secondary schools in Huangpu District of Guangzhou. Students 'reports and investigators' daily records were used to evaluate the intervention measures.

RESULTSDuring the one-year period of intervention (Dec. 2004 through Dec. 2005), eighteen activities had been implemented among students, teachers, parents and cigarette retailers. Ranked by the rate of awareness, the top six activities were shown as follows: "the nicotine toxicity experiment" (90.7%), "agreement of building families free of smoking"(77.7%), "a letter to parents" (77.1%), "no-smoking signs" (76.5%), "Blackboard information about health and smoking" (75.0%), and "signature on the 18th World Day of No Smoking" (70.2%). Among all the activities, "the nicotine toxicity experiment" had the highest rate of participation (88.5%), followed by "a letter to parents" (73.6%), "agreement of building families free of smoking" (69.8%), "health education through experiments"(68.6%), "health education through multimedia" (65.7%) and "signature on the 18th World Day of No Smoking" (65.6%). The top seven activities in which students showed greatest interests were "the nicotine toxicity experiment" (64.5%), "signature on the 18th World Day of No Smoking" (33.0%), "health education through experiments" (31.2%), "health education through multimedia" (29.8%), "class meetings with a thesis of smoking" (26.8%), "health pamphlets" (26.6%), "specific textbooks" (25.9%). The extent of students' general satisfaction to the work of tobacco control in school during the last year was 52.4%. The biggest perceived shortcoming for the intervention plan was the low participation of students.

CONCLUSIONSome intervention measures had not been fully carried out among the students and only covered part of them. It is necessary to adjust the previous intervention measures through keeping the nicotine toxicity experiment, health education through multimedia and other measures with extensive participation of students and at the same time, to avoid literal materials, exhibition boards and traditional single-way health education program.

China ; Health Education ; Health Promotion ; Humans ; Smoking Prevention ; Students

The purpose of this study was to explore the expression characteristics of SDF-1 receptor, CXCR4, in mesenchymal stem cells (MSC) of different passages derived from human umbilical cord (hucMSC). The hucMSC were isolated from Wharton's jelly tissue of human umbilical cord by tissue culture. The expressions of specific marker in hucMSC were detected by flow cytometry. The adipogenic and osteogenic induction of hucMSC were detected by alizarin bordeaux and Oil red O staining. The expressions of CXCR4 protein in hucMSC of 2nd-5th passages were detected by flow cytometry, and cxcr4 mRNA levels in hucMSC of 2nd-5th passages were evaluated by real-time quantitative PCR. The results showed that the expression of CD44, CD13, CD71 were positive while CD38, CD117, HLA-DR were negative. After induced by osteogenic and adipogenic inductors, the lipid droplets and calcium nodals appeared in hucMSC, hucMSC stained with oil red O and alizarin red were shown to be positive. The cxcr4 was found in hucMSC of 2nd-5th passages, and their expressions were (89.82 ± 0.62)%, (86.87 ± 1.32)%, (80.50 ± 4.46)%, (70.10 ± 0.68)% respectively. The cxcr4 mRNA was found in hucMSC of 2nd-5th passages, and expression of cxcr4 of 3rd-5th passages were 0.5585 ± 00875, 0.6205 ± 0.1377, 0.4634 ± 0.0447 times of expression of 2nd passage respectively. It is concluded that the cxcr4 mRNA expresses in hucMSC of 2nd-5th passages, and declines when the number of passages increases. Compared with 2nd passage, cxcr4 mRNA levels in hucMSC of 3rd-5th passages decline, but the expression level of cxcr4 mRNA between hucMSC of 3rd-5th passages is stable.
Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Receptors, CXCR4 ; metabolism ; Umbilical Cord ; cytology ; metabolism

OBJECTIVETo investigate the interactive effect of job task and psychosocial factors on the outcomes of musculoskeletal disorders.

METHODS653 workers from different type of manufacturing industries and administration office recruited in a cross-sectional epidemiological survey. The Quick Exposure Check (QEC) was applied to assess the ergonomic load of job task, Job Content Questionnaire (JCQ) for identifying psychological characteristics, and Nordic Standardized Questionnaire for investigating outcomes of WMSDs.

RESULTSThe prevalence of WMSD in shoulder, upper back, lower back and hand/wrist were significantly different under a variety of combined job task and psychosocial characteristics (P < 0.05 or P < 0.01). The more physical and psychological loads, the higher prevalence of WMSDs were revealed. By using multivariate analyses, a potential interactive effect was found in terms of the WMSDs symptoms in hand/wrist, shoulder, upper back and lower back after adjusted by work year, age, and gender.

CONCLUSIONSHigher physical load and greater psychosocial risk are more frequent self-reported symptoms of WMSDs than those of lower exposures. Ergonomic intervention strategies aimed at reducing the incidence of WMSDs should not only be focused on control of physical work factors but also psychosocial risks of relevance.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Musculoskeletal Diseases ; etiology ; psychology ; Occupational Diseases ; etiology ; psychology ; Stress, Psychological ; Surveys and Questionnaires ; Task Performance and Analysis ; Young Adult

BACKGROUNDThe morphological and electrophysiological characteristics of cardiac cells in Koch triangle are still disputed. We studied the appearance and electrical properties of these diverse myocytes to elucidate their complex electrophysiological phenomena.

METHODSExperiments were conducted using cooled charge coupling device (CCD) system and whole cell, patch clamp technique to determine the morphology, action potential and sodium current density of single viable myocytes enzymatically isolated from the Koch triangle of rabbit hearts.

RESULTSMorphologically, cardiac cells in shape of spider, tiny spindle, slender spindle, rod and strip were observed in percentage of 3.0 +/- 0.3, 35.0 +/- 5.0, 15.0 +/- 2.0, 40.0 +/- 5.0 and 6.0 +/- 0.7 respectively. The cellular dimensions and capacitance gradually increased in the above order (all P < 0.05). Electrophysiologically, action potential configurations recorded from them were similar respectively to nodal (N), atrial nodal (AN), nodal Hisian (NH), atrial (A) and Hisian like potentials obtained from the intact atrioventricular nodal preparations. Diastolic depolarization appeared in all myocytes except for rod cells. Sodium current density increased in the order of tiny spindle, strip, rod, slender spindle cell (all P < 0.05), but could not be detected in spider-shaped cells. Linear regression analysis revealed that membrane capacitance was correlated negatively to the rate of diastolic depolarization r = -0.70, P < 0.001, but positively to maximum depolarization potential, amplitude of action potential, upstroke velocity and maximum peak value of sodium current density r = 0.84, 0.80, 0.87 and 0.75, respectively; all P < 0.001.

CONCLUSIONSThe results demonstrated that spider-shaped, spindle, rod and strip cells in Koch triangle might correspond to pacemaking, transitional, atrial and Purkinje like cells, respectively. Furthermore, tiny spindle and slender spindle cells were referred to transitional cell alpha (TCalpha) and beta (TCbeta) accordingly considering their distinctive electrical properties. Different myocytes with diverse electrical properties constituted the infrastructure of sophisticated electrophysiological phenomena in Koch triangle. In view of the prominent percentage and electrical properties, tiny spindle and slender spindle cells were presumed to play important roles.

Action Potentials ; Animals ; Atrioventricular Node ; cytology ; physiology ; Female ; Male ; Myocytes, Cardiac ; cytology ; physiology ; Rabbits ; Sodium Channels ; physiology