The liquid-chromatography-mass technology was used in the metabolomic analysis of mouse's blood 1 hour after exhaustive exercise,in order to explore the potential mechanism of Rhodiola in fatigue elimination of exhaustive exercise mouse.The exhaustive mouse model was made by loaded-swimming.A total of 24 mice were randomly divided into theRhodiola + exercise group,exercise group andno-exercise group.The dose of Rhodiola was 0.4375 g·kg-1·d-1.The loaded-swimming was conducted after two successive weeks of medication.Blood was collected 1 h after swimming for the sample preparation.The enzyme assay and anthrone colorimetry were used to test blood lactate acid and glucose,respectively.UFLC-Q-TOF was used to detect metabolic profiles of each group.The principal component analysis (PCA),orthogonal partial least squares discriminant analysis (OPLS-DA) and heat map analysis were used to compare differences among groups with score chart and to obtain the characteristics biomarkers by load chart.The results showed that the blood lactate acid level of theRhodiola+ exercise group was significantly lower than that of theexercise group.And the glucose level of theRhodiola+ exercise group was significantly higher than that of theexercise group.The metabolomic analysis showed that there were no obvious changes on 1,25-(OH)2D3,diacylglycerol (DG) and inositol triphosphate (IP3).All three materials in theRhodiola + exercise group were significantly lower than those of theexercise group.They were much closer to theno-exercise group.And all three materials were related to the increasing of muscle tension.It was concluded that Rhodiola had the function of promoting fatigue eliminating.This effect may be related to cell membrane protection,regulation of 1,25-(OH)2-D3→IP3,DG pathway,and relieving of muscle tension after exercises.

Objective To evaluate the role of μ-opioid receptor (MOR) in attenuation of bone cancer pain by anti-nerve growth factor (anti-NGF) in rats. Methods Part Ⅰ Sixty female SD rats weighing 200-220 g were randomly divided into 4 groups (n = 15 each): sham operation group (group S), sham operation + anti-NGF group (group SN), bone cancer pain group (group P) and bone cancer pain+ anti-NGF group (group PN) . Bonecancer was induced by intra-tibial inoculation of 1 × 105 Walker 256 breast cancer cells in group P and PN. Group S and SN received injection of PBS 10 μl. APE 10 catheter was inserted at L2,3 interspace into the epidural space 13 days after cancer cell inoculation. Three days after the catheter was successfully placed, group SN and PN received intrachecal (IT) injection of anti-NGF 10 μg (in normal saline (NS) 10 μl) and group S and P IT injection of NS 10 μl twice a day for 5 consecutive days. The number of spontaneous flinches (NSF), paw withdrawal latency (PWL) and paw withdrawal threshold (PWT) were measured before and 13, 16, 18, 21 day after cancer cell inoculation. The animals were sacrificed at 21 day after cancer cell inoculation and the spinal cord dorsal horn and dorsal root ganglion were removed for determination of MOR and MOR mRNA expression. Part Ⅱ Thirty female SD rats weighing 200-220 g were randomly divided into 2 groups (n = 15 each): bone cancer pain + anti-NGF group (group PN) and bone cancer pain + naloxone + anti-NGF group (group PNN). Bone cancer was induced by intratibial inoculation of 1 × 105 Walker 256 breast cancer cells. APE 10 catheter was inserted at L2-3 interspace into the epidural space 13 days after cancer cell inoculation. Three days after the catheter was successfully placed,group PN received IT injection of anti-NGF 10 μg (in NS 10 μl) and group PNN IT injection of naloxone 10μg (in NS 25 μl) and 0.5 h later IT injection of anti-NGF 10 μg (in NS 25μl) twice a day for 5 consecutive days. NSF,PWL and PWT were measured before and 13, 16, 18, 21 days after cancer cell inoculation. Results Part ⅠCompared with group S, no significant change was found in NSF, PWL and PWT in group SN, and in MOR and MOR mRNA expression in group SN and PN (P ＞ 0.05), NSF was significantly increased, PWL shortened, PWT decreased at 13-21 days after inoculation in group P and PN, and MOR and MOR mRNA expression was down-regulated in group P (P ＜ 0.05 or 0.01). Compared with group P, NSF was significantly decreased, PWL prolonged, PWT increased, MOR and MOR mRNA expression was up-regulated in group PN at 18-21 days after inoculation (P ＜ 0.05 or 0.01). Part Ⅱ Compared with group PN, NSF was significantly increased, PWL shortened, PWT decreased at 18-21 days after inoculation in group PNN (P ＜ 0.05 or 0. 01). Conclusion The mechanism by which anti-NGF attenuates bone cancer pain in rats is related to the activation of MOR.

BACKGROUND: To search for an alloxenogeneic bone with good load bearing function and osteoblastic activity for treating bone defects is an important study issue. We have made a comparative study on its biome chanical characteristics and found that there was no significant difference in maximum load stress, maximum pressure as compared with fresh bone of the same size. Clinicians are concerned about the osteoblastic activity and whether the osteoblastic activity can be reserved after human allogenous a cellular bone (HAB) loaded with bone marrow stromal cells (BMSCs). OBJECTIVE: To investigate the experimental effect of HAB loaded with induced BMSCs, and observe the cellular adherence and growth as well as detect its osteoblastic activity. DESIGN: Single sample experiment. SETTING: Second Affiliated Hospital of Harbin Medical University. MATERIALS: This experiment was conducted at the Experimental Center of the Second Affiliated Hospital of Harbin Medical University between January 2003 and August 2004. HAB was obtained from fresh corpse iliac bones (donated voluntarily). METHODS: Connective tissues and cell compounds of the iliac bones were removed by processing with hydroperoxide andether solution and sterilized for preparing HAB. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium containing DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascor bic acid. Proliferation and differentiation of bone stromal cells were deter mined by detecting the level of alkaline phosphatase (ALP) and osteocalcin (OCN) in the culture medium. Induced bone stromal cells solution was condensed and implanted within HAB scaffold. Cellular osteoblastic activ ity was determined through morphological observation under the light mi croscope and electron microscope as well as biochemical index detection. MAIN OUTCOME MEASURES: ① Detection results of ALP and OCN of BMSCs/HAB composite. ② Histological observation results of BMSCs/ HAB composite. RESULTS: ① Iliac bone block cells were cleaned with good reservation of bone matrix. ② The level of ALP and OCN of MSCs was higher after in ducing for 8 days than that in control group [MSCs after induction: (181.54±40.01) nkat/L, (7.2±1.3) μg/L. There was no method to detect the level in control group, P ＜ 0.05]. ③ BMSCs were adhered and grew well in HAB scaffold. CONCLUSION: HAB loaded with induced BMSCs has an excellent os teogenic function in vitro and shows an effective potential as a good bone tissue engineering material.

Objective To evaluate the effects of dexmedetomidine (DEX) on cell apoptosis induced by endoplasmic reticulum stress and c-Jun N-terminal kinase (JNK) pathway during one-lung ventilation (OLV) in rats.Methods Sixty male Sprague-Dawley rats were randomly allocated into 6 groups (n =10 each):sham operation group (Sham group),OLV group,OLV + atipamezole (α2 receptor antagonist) group (AD group),OLV + atipamezole + DEX group (DEX+AD group),OLV + low-dose DEX group (DEX-L group) and OLV + high-dose DEX group (DEX-H group).The animals were anesthetized with 10％ chloral hydrate 4.5 ml/kg,tracheostomized and mechanically ventilated.Bilateral lungs were ventilated for 2.5 h in Sham group.The right lung was ventilated for 2.0 h followed by 0.5 h two-lung ventilation in OLV group.In DEX-L and DEX-H groups,DEX was infused intravenously for 1 h at a rate of 2.5 μg · kg-1 · h-1 and 5.0 μg · kg-1 · h-1,respectively,starting from 1 h prior to OLV.Atipamezole 250 μg/kg was injected intravenously at 1 h prior to OLV in AD group.Atipamezole 250 μg/kg was injected intravenously at the onset of DEX infusion (5.0 μg · kg-1 · h-1) in DEX+AD group.The rats were sacrificed and left lungs were removed for determination of weight to dry lung weight ratio (W/D),cell apoptosis in lung tissues (by TUNEL),and expression of glucose-regulated protein 78 (GRP78) mRNA and protein,JNK mRNA and phosphorylated JNK (p-JNK) protein (by RT-PCR and Western blot).Pathological changes of lungs were examined and the injured alveolus rate (IAR) was counted under light microscope.The changes in ultrastructure of lung tissues were observed under transmission electron microscope.Apoptosis index (AI) was calculated.Results W/D,AI and IAR were significantly higher in OLV,AD and DEX+AD group than in Sham group,while lower in DEX-L and DEX-H groups than in OLV,AD and DEX+AD groups.The pathological changes of the structure of lung tissues were observed in OLV,AD and DEX+AD groups,while the pathological changes were significantly alleviated in DEX-L and DEX-H groups.In OLV,AD and DEX + AD groups,there was apoptosis in lots of pulmonary vascular endothelial cells and alveolar epithelial cells,while cell apoptosis was significantly reduced after administration of DEX.The expression of GRP78 mRNA and protein,JNK mRNA and p-JNK protein was significantly higher in OLV,AD and DEX+AD groups than in Sham group,and lower in DEX-L and DEX-H groups than in OLV,AD and DEX +AD groups.Conclusion DEX pretreatment can protect lungs during OLV,and inhibited JNK signaling pathway and reduced cell apoptosis induced by endoplasmic reticulum stress may be involved in the mechanism.

Aim To explore the proteomics mechanism of the differentiation induction effect of 4-amino-2-trif-luoromethyl-phenyl retinate(ATPR)on human leukemi-a K562 cells. Methods Human leukemia K562 cells were incubated with the same concentration (1 × 10 - 6 mol·L - 1 ) of ATPR or ATRA for 48 hours. The total cell proteins were collected, purified and digested by trypsin, solid phase extraction, and the peptides were detected by ESI-LC-MS / MS. The difference of the pro-tein expression between the cells treated with ATPR and ATRA was compared by using the Discoverer Pro-teome 1. 2 software, and the molecular function, the biological process and other information of those pro-teins were analyzed based on the DAVID, KEGG, STRING databases. Results 120 specific proteins were identified only in the ATPR group, 143 only in the ATRA group, and 422 other proteins in both groups. Results of DAVID analysis showed that ATPR-induced specific proteins were mainly involved in 39 biological processes of proteins and macromolecules metabolism, protein transport and localization and so on. Results of KEGG analysis revealed that ATPR-in-duced proteins participated in signal pathways, mainly metabolic pathways, PI3K-Akt signal pathway, TGF-beta signal pathway and other pathways in cancer. String protein interaction network analysis displayed that ATPR-induced proteins, like EIF3A, EIF6, RPL3, RPL8, RPL13, RPL7A, RPL21, RPS3, RPS14, NACA, BTF3, NHP2L1, PPP2CA proteins had direct interactions with more than or equal to 10 associated proteins. Conclusion The differentiation induction effect of ATPR on K562 cells might be as-cribed to the ATPR-induced proteins interaction net-work and the specific central proteins it induced, which are involved in the regulation of cell prolifera-tion, differentiation and apoptosis.

Objective:To clone and express panallergen profilin from the pollen of coco(Cocos nucifera Linnaeus).Methods:RT-PCR and RACE methods were applied to clone the full-length panallergen genes from coco pollen and the sequence was analyzed.The specific primers were designed.The ORF of profilin of coco pollen was amplified with RT-PCR and cloned into the expression vector pET 28a.Expression of the recombinant coco pollen profilin was carried out in E.coli BL21(DE3) and the purification of the recombinant protein was performed via affinity chromatography with Ni2+ coupled to sepharose.IgE reactivity to recombinant coco pollen profilin was investigated by immunoblot.Results:The complete sequence of coco pollen profilin was cloned.The sequence was 608 bp and included an open reading frame(396 bp) coding for 131 amino acids.Sequence analysis showed that the deduced protein was an acidic protein with an estimated molecular mass of 14.19 kD and a pI of 4.61.The GeneBank accession number of the clones was EF173598.After overexpressed in E.coli BL21(DE3),the recombinant protein was purified through affinity chromatography with Ni2+ coupled to sepharose.Immunoassay showed that the recombinant allergen has good IgE binding capacity.Conclusion:The profilin of coco pollen is expressed successfully in BL21(DE3),which will be used as a base for further study on coco pollen related allergy.

OBJECTIVE:To investigate the efficacy of rosiglitazone in the treatment of patients with nonalcoholic fatty liver(NAFLD).METHODS:A total of 124 patients with NAFLD(excluding those with cirrhosis) were randomly divided into rosiglitaone(RSG) treatment group and placebo group(control) for a treatment of 24wk.Then the curative effects in two groups were compared.RESULTS:As compared with placebo,RSG showed a better efficacy in the improvement of the liver function,levels of blood glucose and blood lipid(P

Objective:To investigate the changes and significances of serum cystatin C and transforming growth factor-β1 levels for the neonatal asphyxia.Methods:Forty-six asphyxia newborns were chosen as the asphyxia group,and thirty healthy newborns were selected as the control group.The TGF-β1,CysC,BUN,Scr,and GFR levels of both groups were detected on the 1st,3rd,7th day after hospitalization.According to the renal injury,the 46 newborns were divided into normal group and asphyxia group,and the serum indexes were detected and analyzed.Results:On the 1st,3rd,7th day after hospitalization,the TGF-β1,GFR of asphyxia group was obviously increased and was lower than those of the control group (P＜0.05);the level of CysC,BUN,Scr in both groups were decreased,and the change degree in asphyxia group were higher than that of the control group (P＜0.05);the CysC,BUN,Scr in renal injured group were higher than those of normal group,and TGF-β1,GFR were much lower (P＜0.05).Additionally,TGF-β1 level of renal injured group was negatively correlated to the BUN and Scr,and positively correlated with the GFR (P＜0.05).The level of serum CysC in renal injured group was positively correlated to BUN and Scr and negatively correlated to GFR (P＜0.05).Conclusion:The serum TGF-β1,CysC in asphyxia newborns had significant changes compared with the healthy newborns and was correlated to the renal injured indexes,which had clinical directive significance on the early diagnosis,condition judgment,and prognosis of neonatal asphyxia with renal injury.

Objective:To investigate the correlation between low-density lipoprotein cholesterol (LDL-C) and hemorrhagic transformation (HT) after intravenous thrombolysis in patients with acute ischemic stroke (AIS).Methods:Patients with AIS received intravenous thrombolysis using standard dose alteplase in the First Hospital of Shanxi Medical University from January 2014 to December 2019 were enrolled retrospectively. Head CT scan was performed within 24 h after thrombolytic therapy to identify the occurrence of HT. The demographic and baseline clinical data were compared between the HT group and the non-HT group. Multivariate logistic regression analysis was used to determine the correlation between LDL-C and HT after thrombolysis. Results:A total of 323 patients with AIS who received intravenous thrombolytic therapy were enrolled, their age was 65±12 years (range, 54-78 years), and 219 were males (67.8%). The median baseline National Institutes of Health Stroke Scale (NIHSS) score was 4 (interquartile range, 3-9). Ninety one patients (28.17%) developed HT, of which 8 (2.48%) had symptomatic intracerebral hemorrhage. Univariate analysis showed that there were significant differences in LDL-C, age, baseline NIHSS score, baseline systolic and diastolic blood pressure, baseline fibrin degradation products, and the proportion of patients with atrial fibrillation and stroke etiology between the HT group and the non-HT group (all P<0.05). Multivariate logistic regression analysis showed that lower LDL-C (odds ratio [ OR] 0.531, 95% confidence interval [ CI] 0.358-0.788; P=0.002), higher baseline NIHSS score ( OR 1.063, 95% CI 1.010-1.120; P=0.020) and higher baseline systolic blood pressure ( OR 1.015, 95 CI 1.004-1.026; P=0.008) were the independent risk factors for HT after intravenous thrombolysis in patients with AIS. Conclusions:Low LDL-C is an independent risk factor for HT in patients with AIS after intravenous thrombolysis. The patients with lower LDL-C should be cautious in lipid-lowering therapy and be alert to the occurrence of HT.