Objective To explore the relationship between muscle fatigue and pain in lumbar muscle fatigue induced test. Methods From July to October, 21 healthy female volunteers were subjected to the fatigue induced test. Their root mean square (RMS) and median frequency (MF) of the surface electromyography were collected, their fatigue were assessed with Borg Fatigue Scale and Visual Analogue Scale (VAS), and their maximal voluntary contraction (MVC) and pressure pain threshold (PPT) were measured, before, during and after ex-periment. They were divided into pain or no-pain groups according to the symptom, and the differences before and after test were compared. Results There was significant difference in the scores of Borg Fatigue Scale and VAS, MVC, RMS, and MF after test (Z>6.064, P<0.001). There was significant difference between two groups in all the data before test, and there was significant difference between two groups in the difference before and after test of the VAS score (t=-4.112, P=0.001) and RMS (t>2.385, P<0.05) in some points. Receiver operating characteristic curve analysis showed that the optimal cut-off point of VAS score difference was 3.45 in predicting the pain onset, with sensi-tivity of 100％and specificity of 81.8％. Conclusion For the fatigue induced test, the fatigue VAS score difference may predict the induced pain, and RMS difference associate with the occurrence of pain. No significant is observed of the Borg Fatigue Scale and PPT.

Cholecystolithiasis with choledocholithiasis (CCL) is a common disease.The removal of common bile stone is a challenge for the surgery.This paper discussed the clinical application of three stone removal techniques including direct stone removal,irrigation and stone extraction by basket under cholangioscopy in order to take the stones effectively and safely,shorten the procedure time,avoid the injuries of common bile duct wall caused by the repetition of a single method such as biliary endoscopic stone extraction,reduce the difficulty of taking stone and enhance recovery of patients.

OBJECTIVETo explore a new method for identification Astragali Radix from its adulterants by using ITS sequence.

METHODThirteen samples of the different Astragali Radix materials and 6 samples of the adulterants of the roots of Hedysarum polybotrys, Medicago sativa and Althaea rosea were collected. ITS sequence was amplified by PCR and sequenced unidirectionally. The interspecific K-2-P distances of Astragali Radix and its adulterants were calculated, and NJ tree and UPGMA tree were constructed by MEGA 4.

RESULTITS sequences were obtained from 19 samples respectively, there were Astragali Radix 646-650 bp, H. polybotrys 664 bp, Medicago sativa 659 bp, Althaea rosea 728 bp, which were registered in the GenBank. Phylogeny trees reconstruction using NJ and UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Astragali Radix from adulterants.

CONCLUSIONITS sequence can be used to identify Astragali Radix from its adulterants successfully and is an efficient molecular marker for authentication of Astragali Radix and its adulterants.

Althaea ; classification ; genetics ; Astragalus membranaceus ; classification ; genetics ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Fabaceae ; classification ; genetics ; Medicago sativa ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Roots ; genetics ; RNA, Ribosomal ; genetics ; RNA, Ribosomal, 5.8S ; genetics ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo study the effect of endothelin-1 (ET-1) and its antagonists on the expression of endothelin and its receptors mRNA in HSC-T6 cells.

METHODSCultured HSC-T6 cells were randomly divided into 7 groups: Sham control group, ET-1 group (10 nmol/L ET-1), BQ-123 group [1 micromol/L BQ-123, a selective endothelin receptor A (ETRA) antagonist], BQ-788 group [1 micromol/L BQ-788, a selective endothelin receptor B (ETRB) antagonist], ET-1 + BQ123 group (10 nmol/L ET-1 + 1 micromol/L BQ-123), ET-1 + BQ-788 group (10 nmol/L ET-1 + 1 micromol/L BQ-788) and ET-1 + BQ-788 group (10 nmol/L ET-1 + 1 micromol/L BQ-123 + 1 micromol/L BQ-788). The expression of endothelin receptor mRNA of HSC-T6 cells was determined by reverse-transcription polymerase chain reaction (RT-PCR).

RESULTSThe expression of ETRA mRNA in ET-1 + BQ123 + BQ788 and ET-1 + BQ788 group was significantly lower than ET-1 group (0.329 +/- 0.044 and 0.292 +/- 0.023 vs. 0.440 +/- 0.030 P < 0.05). Compared with ET-1 group, the expression of ETRB mRNA in ET-1 + BQ788 group was down regulated obviously (0.499 +/- 0.136 vs. 0.153 +/- 0.071, P < 0.05). There was no significant difference in ET-1 + BQ123 group and ET-1 + BQ123 + BQ788 group when compared with ET-1 group (0.499 +/- 0.136 vs. 0.496 +/- 0.103 and 0.299 +/- 0.129, P > 0.05).

CONCLUSIONSET-1 has no obvious effect on the expression of ETRA mRNA in HSC-T6. ET-1 may up-regulate the expression of ETRB mRNA. Act on ETRA receptor, ET-1 can inhibit the expression of ETRB mRNA.

Animals ; Cells, Cultured ; Endothelin-1 ; pharmacology ; Gene Expression Regulation ; drug effects ; Hepatocytes ; drug effects ; metabolism ; Oligopeptides ; pharmacology ; Peptides, Cyclic ; pharmacology ; Piperidines ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptor, Endothelin A ; biosynthesis ; drug effects ; genetics ; Receptor, Endothelin B ; biosynthesis ; drug effects ; genetics

OBJECTIVETo investigate the possibility of transdifferentiation of adipose mesenchymal stem cells (AMSCs) into hepatocytes.

METHODSHuman omentum adipose tissue was dispersed with collagenase I. Cells collected were cultured in a DMEM-F12 medium containing 2% FBS supplemented with 20 ng/ml HGF, 10 ng/ml FGF4, 1xITS and 0.1 micromol/L dexasmison. The cells of the control group were also cultured in the same kind of medium but without any cytokines serving as a control. The expression of hepatic transcriptional factors such as GATA4 and HNF1 were checked by RT-PCR. At the end of the induction, hepatocyte markers were analysed by flow cytometry, and cytokeratin expressions were examined using cyto-immunofluorescence methods.

RESULTSAMSCs grew like fibroblasts and were passaged easily. Most of the third passaged AMSCs were positive against anti-CD29, anti-CD44 antibodies, but negative for the anti-CD34 and anti-CD45 ones. The hepatic transcriptional factor was expressed gradually to higher levels during the induction time. AFP and Alb positive cells were 30.0% and 17.8% of the total cultured cells, and the rate of cells positive to the two markers was 6.9%. The inducted cells were positive for CK18 and CK19 antibodies at the end of the induction. The cells in the control group were negative when checked by these methods.

CONCLUSIONSAMSCs could be directed to differentiate into hepatocytes in vitro by a cytokine cocktail with a low concentration FBS culture system.

Adipocytes ; cytology ; Cell Differentiation ; Cell Transdifferentiation ; Cells, Cultured ; Hepatocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells (ADSCs).

METHODSADSCs were isolated from human abdominal adipose tissue and cultured. The immunophenotype and adipose induced-differentiation were identified, and the third generation cells were collected. The collected cells were assigned to 1 x 10(-8), 1 x 10(-7), 1 x 10(-6) mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs' nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry.

RESULTSThe secretion amounts of VEGF and HGF of ADSCs in 1 x 10(-8) and 1 x 10(-7) mol/L insulin groups [(471 +/- 41, 762 +/- 66 ng/L), (643 +/- 64, 930 +/- 67 ng/L), respectively] were significantly higher as compared with those in control group (286 +/- 47, 577 +/- 84 ng/L) (P < 0.05 or P < 0.01). No change occurred in the secretion amount of VEGF and HGF of ADSCs in 1 x l0(-6) mol/L insulin group (P > 0.05). The supernatant fluid of ADSCs' nutrient medium of 1 x 10(-8), 1 x 10(-7) mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1 x 10(-7) mol/L group (P < 0.05 or P < 0.01).

CONCLUSIONSInsulin in the concentrations of 1 x 10(-8) and 1 x 10(-7) mol/L can notably promote ADSCs' function of secreting VEGF and HGF.

Adipocytes ; cytology ; drug effects ; secretion ; Cells, Cultured ; Fibroblasts ; cytology ; Hepatocyte Growth Factor ; metabolism ; Humans ; Insulin ; pharmacology ; Stem Cells ; cytology ; drug effects ; secretion ; Vascular Endothelial Growth Factor A ; metabolism

Objective To compare the safety and effectiveness of Propofol-Fentanyl and Propofol-Remifentanil total intravenous anesthesia for airway foreign body (FB) removal in children. Method 280 children aged 1 ~ 3 years underwent rigid bronchoscopy for FB removal were randomized into two groups. The Fentanyl group (Group F, n = 140) were given Propofol 2.00~3.00 mg/kg and Fentanyl 2.00 μg/kg for induction and Propofol 200.00 ~ 500.00 μg/(kg·min) for maintenance of anesthesia. The Remifentanil group (Group R, n = 140) were given Propofol 2.00 ~ 3.00 mg/kg and Remifentanil 1.00 ~ 1.50 μg/kg for induction of anesthesia, while anesthesia was maintained with Propofol 200.00 ~ 500.00 μg/(kg·min) and Remifentanil 0.10 ~ 0.20 μg/(kg·min). All the children during the procedure were with spontaneous respiration. SpO2 before inserting rigid bronchoscope (T1), 1 min (T2) and 3 min (T3) after insertion, 3 min (T4) and 10 min (T5) after extraction were recorded. PETCO2 after endoscopy (T6) was measured. Adverse events, including body movement, cough, breath-holding, and hypoxemia,were observed. The time of induction, surgery, recovery and the total dosage of the intravenous agents were recorded. Results SpO2 of the two groups were in normal range at T1 ~ 5, which was higher in group R than group F at T2 ~ 5 (P < 0.05). PETCO2 of group R was lower than group F at T6 (P < 0.05). The rate of body movement and cough were comparable between the two groups (P > 0.05), while breath-holding and hypoxemia were more frequent in group F (P < 0.05). The time of induction and recovery were shorter in group R (P < 0.05), while surgery time and the Propofol dosage were similar (P > 0.05). The total dose of Fentanyl was significantly higher than Remifentanil (P < 0.05). Conclusion Combination of Propofol with Fentanyl or Remifentanil both produce effective anesthesia in children undergoing FB removal. But Propofol-Remifentanil provides more stable oxygen saturation, faster induction and recurrence of anesthesia, as well as less intraoperative complications.

Animals ; Apoptosis ; Female ; Gene Transfer, Horizontal ; Genetic Therapy ; Hemorrhage ; pathology ; therapy ; Hepatocytes ; pathology ; Interleukin-10 ; genetics ; Male ; Pancreatitis, Acute Necrotizing ; pathology ; therapy ; Proto-Oncogene Proteins c-bcl-2 ; physiology ; Rats ; Rats, Wistar ; bcl-2-Associated X Protein

OBJECTIVETo investigate the molecular mechanism of atherosclerosis that related to age.

METHODSImmunohistochemistry staining and Western blot were adopted to determine the nuclear translocation of nuclear factor-kappa B (NF-kappaB) and expression of platelet-derived growth factor B (PDGF-B) in smooth muscle cells (SMCs) co-cultured with low density lipoprotein (LDL), oxidized LDL (ox-LDL), and ox-LDL+high density lipoprotein (HDL) originated from rats of 2 and 10 months old respectively. Fat stain was used to identify the lipid intake in SMCs.

RESULTSThe optimal stimulation time of ox-LDL to SMCs was 12 hours. NF-kappaB intensity increased in most nuclei of SMCs that originated from rats of either 2 or 10 months old co-cultured with ox-LDL. The intensity of NF-KB and the amount of intracellular lipid taken in SMCs were more obvious in cells from 10-month-old rats than from the younger ones. Change of PDGF-B expression in SMCs was not remarkable in each group of rats.

CONCLUSIONSThe 10-month-old rats are more susceptive to ox-LDL than 2-month-old rats in activating nuclear translocation of NF-kappaB. Maybe this is one of the important reasons contributing to the difference between the older and younger rats on the initiation and development of atherosclerosis lesion. Expression of PDGF-B is not associated with the activity of nuclear translocation of NF-kappaB.

Age Factors ; Animals ; Aorta ; cytology ; Cell Nucleus ; metabolism ; Cells, Cultured ; Culture Media ; Lipoproteins, HDL ; pharmacology ; Lipoproteins, LDL ; pharmacology ; Male ; Myocytes, Smooth Muscle ; metabolism ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-sis ; metabolism ; Rats ; Rats, Wistar

Objective To investigate the differences in the expression of vitamin D receptor (VDR) and serum vitamin D levels in subcutaneous adipose tissue between overweight/obese and normal-weight gravidas, and the relationship between these two indicators and gestational diabetes mellitus (GDM). Methods Women with full-term singleton pregnancies who underwent elective cesarean section in Changzhou Maternal and Child Health Care Hospital Affiliated to Nanjing Medical University from January 2015 to April 2017 were enrolled. Among them, there were 70 cases GDM women, including 35 normal-weight (NW-GDM group) and 35 overweight/obese women (OW-GDM group). During the same period, another 70 pregnant women with normal glucose tolerance who underwent scheduled cesarean delivery were selected as the control group, including 35 normal weight women (NW-control group) and 35 obese/overweight women (OW-control group). Fasting blood samples were collected before operation to determine the levels of different biomarkers, including vitamin D, lipid, fasting blood glucose, fasting insulin and adiponectin, and to calculate the homeostasis model assessment-insulin resistance (HOMA-IR). Two subcutaneous adipose tissue samples of the abdominal wall were taken during the operation to detect the expression and distribution of VDR protein with immunohistochemistry. Meanwhile, VDR mRNA transcription level was quantitatively analyzed using real-time fluorescence quantitative polymerase chain reaction. One-way analysis of variance, LSD, Kruskal-Wallis test, Mann-Whitney U test, Chi-square test and logistic regression analysis were used for statistical analysis. ResuLts (1) The body mass index (BMI) of the OW-control group and the OW-GDM group before pregnancy and delivery were all higher than that of the NW-control group and the NW-GDM group [BMI before pregnancy: (29.2±2.9), (29.4±3.8) vs (21.1±2.3) and (21.9±2.0) kg/m2, F=87.766; BMI before delivery: (35.2±3.4), (35.1±4.3) vs (27.9±2.8) and (28.8± 3.3) kg/m2, F=44.827; all P<0.001]. Newborn birth weight and the proportion of diabetic family history in the OW-GDM group were higher comparing to the NW- and OW- control group [(3 893±498) vs (3 501±402) and (3 625±332) g, F=4.751; 22.9%(8/35) vs 5.7%(2/35) and 5.7%(2/35), χ2=7.869; all P<0.05]. (2) In the OW-control group, the fasting insulin level and HOMA-IR were higher and the adiponectin and vitamin D concentration were lower than those in the NW-control group [13.3(12.3-14.5) vs 12.0(10.4-13.3) mmol/L, 2.7(2.4-3.0) vs 2.2(2.0-2.7), (61.8±20.4) vs (74.9±29.3) ng/ml, (21.6±7.2) vs (25.9±7.3) ng/ml; all P<0.05], and similar results were found between the OW-GDM group and the NW-GDM group [15.3(12.3-19.5) vs 12.0(10.1-15.8) mmol/L, 3.4(2.6-4.1) vs 2.6(2.1-3.2), (50.3±22.3) vs (62.1±23.2) ng/ml, (17.1±6.7) vs (20.6±7.9) ng/ml, all P<0.05]. Compared with the NW-control group, the NW-GDM group had higher fasting glucose and lower high density lipoprotein-cholesterol (HDL-C), adiponectin and vitamin D levels [4.6(4.3-5.1) vs 4.3(4.0-4.5) mmol/L, 1.7(1.6-1.9) vs 2.1(1.6~2.4) mmol/L, (62.1±23.2) vs (74.9±29.3) ng/ml, (20.6±7.9) vs (25.9±7.3) ng/ml; all P<0.05]. Compared with the OW-control group, fasting glucose, fasting insulin and HOMA-IR were higher and HDL-C, adiponectin and vitamin D levels were lower in the OW-GDM group [4.7(4.4-5.4) vs 4.5(4.2-4.7) mmol/L, 15.3(12.3-19.5) vs 13.3(12.3-14.5) mmol/L, 3.4(2.6-4.1) vs 2.7(2.4-3.0), 1.6(1.4-1.8) vs 1.9(1.7-2.2) mmol/L, (50.3±22.3) vs (61.8±20.4) ng/ml, (17.1±6.7) vs (21.6±7.2) ng/ml; all P<0.05]. (3)The overall vitamin D deficiency rate during the third trimester of the four groups was 78.6% (110/140), and the figure was 62.8% (22/35), 82.8% (29/35), 77.1% (27/35) and 91.4% (32/35) in the NW-control group, OW-control group, NW-GDM group and OW-GDM group (χ2=8.994, P=0.029), indicating a higher rate in the OW-GDM group than that in the NW-control group (χ2=8.102, P=0.004). (4) VDR was expressed in the nucleus of adipose tissue in all samples and statistic difference in protein expression was found among the four groups. VDR mRNA expression was higher in both GDM subgroups than that in the two control subgroups, and also higher in the two overweight/obese subgroups than in the corresponding normal-weight subgroups. (5)Serum vitamin D level was negatively correlated with fasting blood glucose and pre-pregnancy BMI, and positively correlated with adiponectin (P<0.05). The incidence of GDM was related to family history of diabetes, VDR mRNA, total cholesterol, HDL-C and HOMA-IR. ConcLusions GDM and overweight/obese patients had decreased serum vitamin D level and increased VDR in subcutaneous adipose tissue. These two factors are closely related to GDM.