Previous studies have shown that ginsenoside Rb1 (Rb1), one of active components in ginseng, can activate insulin signaling pathway and promote translocation of glucose transporters (GLUTs) to increase glucose uptake in adipocytes. However, the effect of Rb1 on the expressions of GLUTs remains unknown. In this study, the effects of Rb1 on GLUT1 and GLUT4 were observed in 3T3-L1 adipocytes and epididymal adipose tissue of db/db obese diabetic mice. Male db/db mice were treated with Rb1 by intraperitoneal injection at the dosage of 20 mg x kg(-1) for 14 d. Rb1 reduced HOMA-IR significantly (P < 0.05, n = 5), and FBG and FINS sowed declining trend after treatment with Rb1. Rb1 recovered the expressions of GLUT1 and GLUT4 and phosphorylation of AKT in adipose tissue of db/db mice. In vitro, glucose consumption in 3T3-L1 adipocytes treated with 10 micromol x L(-1) Rb1 for 24 h was elevated (P < 0.05, n=3), and mRNA of GLUT1 and GLUT4 were up-regulated (P < 0.05, n=3) and proteins of GLUT1 and GLUT4 were also increased. AKT was activated in adipocytes treated with Rb1 for 3 h. It can be concluded that ginsenoside Rb1 can up-regulate the expression of GLUTs in adipose tissue, in addition to activate insulin signalling pathway, which may partially account for its insulin sensitizing activity and regulating effect of glucose metabolism.
3T3 Cells ; Adipocytes ; drug effects ; Animals ; Cell Line ; Diabetes Mellitus, Experimental ; metabolism ; Ginsenosides ; pharmacology ; Glucose ; metabolism ; Glucose Transport Proteins, Facilitative ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred NOD ; Up-Regulation ; drug effects

OBJECTIVETo construct the acid-sensitive potassium hannel-3(TASK3) eukaryotic expression plasmid and to establish a stable SH-SY5Y cell line expressing enhanced green fluorescent protein (EGFP)-tagged TASK3.

METHODSTASK3 coding region was subcloned into pEGFP-N1 plasmid to construct a recombinant vector alled pEGFP-TASK3. The correct recombinant expressing plasmid was transfected with X-feet transfection reagent to SH-SY5Y cells. The cell line stably expressiing EGFP tagged-TASK3 gene was established by screening with antibiotic G418 and fluorescence microscope. The expression and localization of the EGFP tagged-TASK3 fusion protein was detected by Western blot and confocal microscope. Exposure of the SH-SY5Y cell line expressing stably TASK3-eGFP fusion proteins was exposed to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability was assessed with cell counting Kit-8 (CCK-8).

RESULTSAll the results of identification by PCR, digestion with restriction endonuclease and sequencing indicated that the recombinant eukaryotic expression plasmid pEGFP-TASK3 was constructed correctly. The stable SH-SY5Y cell line expressing EGFP tagged-TASK3 fusion protein was successfully established. Exposure of the wild type SH-SY5Y cells and the stable SH-SY5Y-GFP tag-TASK3 cell line to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability of two group cells significantly reduced with pH declining, and the difference was statistically significant (P < 0.05). Compared with wild type SH-SY5Y cells, the cell viability of stable SH-SYSY-GFP tag-TASK3 cell line increased significantly with the same pH media, and the difference was statistically significant (P < 0.05).

CONCLUSIONThe eukaryotic expression vector pEGFP-TASK3 is successfully constructed and the cell line stably expressing TASK3-eGFP fusion is established which is important for their fundamental research and potential applications.

Blotting, Western ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; Polymerase Chain Reaction ; Potassium Channels, Tandem Pore Domain ; genetics ; Transfection

Objective To compare the intubating conditions between dexmedetomidine and remifentanil when combined with sevoflurane-nitrous oxide (N2O) for anesthesia induction in the pediatric patients.Methods A total of 122 pediatric patients,aged 4-10 yr,of American Society of Anesthesiologists physical status Ⅰ,undergoing elective plastic surgery,were randomly divided into dexmedetomidine group (group D,n =61) and remifentanil group (group R,n=61).Eight percent sevoflurane and 60％ N2O were inhaled for induction of anesthesia,and the fresh gas flow was set at 6 L/min.After disappearance of eyelash reflex,dexmedetomidine 1 μg/kg and remifentanil 1 μg/kg were intravenously injected over 50-60 s in D and R groups,respectively,and 1 min later tracheal intubation was performed.The intubating conditions were graded,and the satisfactory intubating conditions and successful intubation were recorded.The development of adverse cardiovascular reactions and complications such as hyoxemia and laryngospasm before and after intubation and postoperative pharyngodynia was recorded.Results Compared with group D,no significant change was found in the success rate of intubation,rate of satisfactory intubation,intubating condition grade or incidence of postoperative pharyngodynia (P＞ 0.05),and the incidence of hypertension and sinus tachycardia after intubation was significantly increased in group R (P＜0.05).No pediatric patients developed hyoxemia,laryngospasn or sinus tachycardia in two groups.Conclusion When 8％ sevoflurane and 60％ N2O are inhaled for anesthesia induction,combing with dexmedetomidine 1 μg/kg produces better clinical efficacy than combing with remifentanil 1 μg/kg in improving the intubating conditions for pediatric patients.

Objective To establish a network laboratory for blood cell analysis and better calibrate haematology analyzers in local lab.Methods According to GB/T 15481《General requirements for the competence testing and calibration laboratories》(idt ISO/IEC 17025),we established a network laboratory providing traceability for blood cell analysis.Complete blood count was traced to Calibration Laboratory in NCCL;The secondary standard haematology analyzer with the same model and calibrator with same lot number were used for verification for a long period.Fresh blood from healthy people was used to calibrate haematology analyzers.Results Gradually we have improved our laboratory quality management system, precision as well as accuracy,which was satisfactory.The unified blood sample was adopted to calibrate different equipments in our hospital and showed consistence when compared with calibration analyzer.The correlation coefficient of all tests is more than 0.99.The relative deviation of WBC,RBC,HCT,HGB and PLT are within?7%,?3.5%,?4%,?3% and?15%,respectively.Conclusions Secondary standard systems provides good comparable results with calibration laboratory.Its tracing mode and quality control scheme could ensure the traceability and accuracy of completed blood count.Furthermore,using elective fresh blood from healthy people,the comparable results from different analyzers were achievable.

BACKGROUND: Peripheral nerve injury can lead to extensive changes in central nervous system, and exercise training can promote the recovery of locomotor function following central nervous system injury. OBJECTIVE: To observe the changes of locomotor function and the expression levels of vesicular glutamate transporter VGLUT1 in the spinal cord in a rat model of dorsal root ganglion resection after treadmill exercise and to explore the effect of treadmill training on the locomotor function after peripheral nerve injury. METHODS: Thirty-nine 10-week-old male Wistar rats were randomized into experimental (n=15), control (n=15) and sham operation (n=9) groups. The rats in the experimental and control groups received the dorsal root ganglion resection at L3and L4segments to establish the model of peripheral nerve injury under local anesthesia, while the rats in the sham operation group were only subjected to dorsal root ganglion exposure. The rats in the experimental group underwent 15 m/minute treadmill training at postoperative 7 days, while rats in the other two groups were in free movement. Gait analysis was performed at preoperative 3 days, postoperative 7, 14, 21, and 28 days, respectively, and the behavioral changes of rats were observed. The tissue sections were obtained from L3segment at postoperative 7, 14, and 28 days to detect the expression levels of VGLUT1 in the spinal cord by immunohistochemistry. RESULTS AND CONCLUSION: The peroneal nerve function index in the experimental and control groups was lower than that before surgery and that in the sham operation group at postoperative different time points (all P ＜ 0.05). The index in the experimental and control groups was the lowest on day 7 postoperatively (P ＜ 0.05), then the index gradually increased, but was still lower than the preoperative level (P ＜ 0.05). The index in the experimental group was significantly higher than that in the control group at postoperative 21 and 28 days (P ＜ 0.05). The expression levels of VGLUT1 in the lamina IX in the experimental and control groups were significantly lower than those in the sham operation group at different time points after surgery (P ＜ 0.01). The levels in the experimental group were significantly higher than those in the control group at postoperative 14 and 28 days (P ＜ 0.05). The levels in both groups on a decline after surgery, especially the control group (P ＜ 0.05). These results suggest that treadmill can promote the recovery of locomotor function post peripheral nerve injury.

OBJECTIVETo study the expression of gene associated with retinoid-interferon-induced mortality-19(GRIM-19) in preimplantation embryo of mice and explore its role in embryonic development.

METHODSThe protein and mRNA expressions of GRIM-19 in 2-cell, 4-cell, 8-cell, morula, and blastocyst phases of mice preimplantation embryo were detected by Western blot analysis and Real-time polymerase chain reaction (PCR).

RESULTSGRIM-19 was continuously expressed in every stage of preimplantation embryo of mice. Western blot analysis and Real-time PCR demonstrated a gradual increase of GRIM-19 expression from 2-cell, which reached a peak in 8-cell phase and then decreased progressively.

CONCLUSIONSThe expression of GRIM-19 in mouse preimplantation embryos changes as at different developmental phases. GRIM-19 may play an important role during embryonic development.

Animals ; Blastocyst ; drug effects ; metabolism ; Female ; Interferons ; pharmacology ; Mice ; NADH, NADPH Oxidoreductases ; genetics ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; Tretinoin ; pharmacology

PURPOSE: It aimed to explore the degree of difficulty of caries-detection-associated-topics perceived by dental students and lecturers as pedagogical step in the development of learning objects for e-learning. METHODS: A convenience sample comprising ninety-eight subjects from different academic levels (undergraduate/graduate students and pediatric dentistry lecturers) participated. Two spreadsheets (isolated/relative) were created considering key topics in the caries detection process. The isolated evaluation intended to explore each topic in an isolated way, while the relative intended to classify, comparatively, the participants' perceived difficulty per topic. Afterwards, data were analyzed. All values on spreadsheets were combined obtaining the subject's final perception. Associations between the subjects' degree of the perceived difficulty and academic level were estimated. ANOVA was used to determine differences regarding the perception among evaluated topics in distinct groups. RESULTS: Caries histopathology and detection of proximal carious lesions were the topics perceived as the most difficult in the process of caries detection by both students and lecturers. Differentiation between an extrinsic pigmentation and a brown-spot (caries lesion) as well as differential diagnosis between caries and enamel developmental defects or non-carious lesions were considered as more difficult by undergraduates in comparison to graduates/lecturers (regression-coefficient=14.54; Standard Error=3.34; P<0.001 and 8.40, 3.31, and 0.01 respectively). CONCLUSION: Topics as histopathology and detection of proximal caries lesions were identified as the most difficult despite the academic level. However, some topics are differently perceived according to the group. These results are useful for developing pedagogical material, based on the students real learning needs/expectations.
Brazil* ; Dental Enamel ; Diagnosis, Differential ; Education, Dental ; Humans ; Learning* ; Pediatric Dentistry ; Pigmentation ; Students, Dental

ObjectiveTo observe the thyroid status of pregnant women during the 1st and 2nd trimester of gestation,and its role in brain and neural development of their offspring's.MethodsFrom 2008 to 2009,pregnant women from nine townships of two counties in Wushi and Baicheng in Aksu prefecture of Xinjiang were selected as research subjects according to the survey standard.After informed consent signed,their urinary iodine,serum thyroid stimulating hormone(TSH) and free thyroxin(FT4) were analyzed.The value of thyroid hormone of normal pregnant women was used in diagnosis of subclinical hypothyroidism and hypothyroxinemia in pregnant women.From 2010 to 2011, The brain and neural development status among offspring born by those pregnant women were evaluated with DDST.In accordance with the results of Denver Development Screen Test (DDST) screening,pregnant women were classified into survey and control groups,and the survey group was the suspicious and abnormal of the result of DDST screening(delay),the control group was normal of the result.According to gestational age,pregnant women were divided into 4 gestation groups:G1(6 to 13 weeks),G2(14 to 18 weeks),G3 ( 19 to 23 weeks) and G4(24 to 28 weeks).ResultsA total of 396 cases of pregnant women during the 1st and 2nd trimester of gestation were investigated(survey group 102 cases,control group 294 cases).The median value of urinary iodine concentration among pregnant women in survey group was 152.4 μg/L The prevalence of subclinical hypothyroidism and hypothyroxinemia among pregnant women was 10.78％(11/102) and 3.93％(4/102),respectively.In control group,the median value of urinary iodine concentration was 180.0 μg/L The prevalence of subclinical hypothyroidism and hypothyroxinemia among pregnant women was 7.48％ (22/294) and 4.42％ (13/294),respectively.During the pregnant period from G1 to G3,the median serum TSH of pregnant women in DDST survey group (2.24,3.49,2.85 mU/L) was higher than that of DDST control group( 1.59,2.70,2.28 mU/L).Especially,the difference of TSH between the two groups during the period of G3 was statistically significant (t =4.906,P ＜ 0.05 ).ConclusionsHypothyroidism tendency of pregnant women during the period from gestation week 19th to 23rd may be an important factor in the development of brain abnormalities of their offsprings.

Objective To assess the effectiveness of nocturnal splinting on carpal tunnel syndrome (CTS) by clinical scores and nerve conduction studies (NCS), and explore their correlations.Methods Forty-one patients (64 wrists), chosen from 66 consecutive patients with CTS from April 2009 to January 2010 meeting the inclusion criteria, were enrolled. The enrolled subjects were clinically evaluated by symptom severity scale (SSS) and functional status scale (FSS), and electrophysiologically evaluated by conventional nerve conduction studies (NCS); distal motor latency (DML) of wrist-abductor pollicis brevis, sensory conduction velocity (SCV) of wrist-index finger and wrist-ring finger, and the differences of distal sensory latency between the median and ulnar nerves (△DSL) were measured. The patients were instructed to use each splint on dorsal and palmar surface of the hand, centered at the distal wrist crease, to fix the wrist in neutral posture at bedtime. SSS, FSS and NCS were evaluated before splinting and (3.03±1.16) months after splinting; the follow-up was completed in 29 patients (31 wrists).Results (1) The abnormality rates of DML, wrist-index finger SCV, wrist-ring finger SCV and△DSL were 85.9%, 78.1%, 81.3% and 96.9%, respectively. (2) The SSS scores (1.55±0.38), FSS scores (1.40±0.27) and△DSL (1.24±0.61) after splinting was significantly decreased as compared with those before splinting (1.77±0.38, 1.53±0.31, 0.97±0.60); and the DML [4.14±0.76 (ms)] after splinting was significantly shortened as compared with that before splinting [4.53±1.25 (ms)]. No improvement of clinical scores was noted in 9 patients (14 wrists, 45.8%) after splinting. (3) The SSS scores were less significantly correlated to DML (r=0.420, P=0.019), wrist-index finger SCV (r=-0.425, P=0.017),wrist-ring finger SCV (r=-0.519, P=0.003), and no correlation between SSS scores and△DSL was noted (r=0.189, P=0.309); no correlation between FSS scores and the parameters of NCS was found either (P＞0.05). Conclusions Splinting is effective at least in a short-term in more than halfpatients with CTS.Little correlation is noted between clinical scores and NCS, suggesting that utilization both approaches to assess the therapeutic effect is of more significance. △DSL is the most sensitive parameter in the electrodiagnosis of CTS.

OBJECTIVETo examine plasma adiponectin (ADPN) and tumor necrosis factor-alpha (TNF-alpha) levels and their correlation in children with obesity in order to investigate the roles of both in the development of childhood obesity.

METHODSOne hundred and forty-seven children with obesity and 118 normal children who were randomly sampled from five primary schools from the Kaifu District in Changsha were enrolled. Physical shape indexes, including height, weight, waist circumference, hip circumference, and waist to hip ratio (WHR) were measured. Body mass index (BMI) was calculated. Blood pressure was measured. Percentage of body fat (%BF) was measured with dual energy X-ray absorptiometry. Plasmal levels of ADPN and TNF-alpha were detected using ABC-ELISA. Blood concentrations of triglycerides (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were measured by automatic biochemistry analyzer. Fasting blood glucose level was measured by glucose oxidase method. Fasting blood insulin level was assayed by radioimmunity. Homeostasis model assessment for insulin resistance (HOMA-IR) was performed.

RESULTSPlasma ADPN levels in obese children significantly decreased compared with those in normal children (8.12+/-2.54 mg/L vs 12.22+/-4.68 mg/L; p<0.05), and had a negative correlation with plasma TNF-alpha levels, BMI, WHR and HOMA-IR (p<0.01), and with %BF, fasting insulin, systolic blood pressure and TG (p<0.05). Plasma TNF-alpha levels in obese children significantly increased compared to normal children (171.38+/-34.33 ng/L vs 91.07+/-21.60 ng/L; p<0.01) and positively correlated with BMI, WHR, %BF, fasting insulin, HOMA-IR, TG and systolic blood pressure (p<0.01), and negatively with HDL (p<0.05). Multiple stepwise regression analysis showed that ADPN, BMI and TNF-alpha were main influential factors for %BF (R2=0.926, p<0.01). There was a significant interaction between ADPN and TNF-alpha (p<0.05).

CONCLUSIONSPlasma ADPN levels decreased and plasma TNF-alpha levels increased in children with obesity and both were main influential factors for %BF in children. There was an interaction between ADPN and TNF-alpha, suggesting that they both participate in the development of childhood obesity.

Adiponectin ; blood ; Adolescent ; Blood Pressure ; Body Mass Index ; Child ; Cholesterol, HDL ; blood ; Female ; Humans ; Insulin Resistance ; Male ; Obesity ; blood ; etiology ; Regression Analysis ; Tumor Necrosis Factor-alpha ; blood