Objective To investigate the possibility of differentiation of multipotent adult progenitor cells from human bone marrow(hMAPCs) into hepatocytes with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro to find a new cell source for liver tissue engineering. Methods (1) Harvest of the hMAPCs: The bone marrow mononuclear cells were obtained from bone marrow obtained from volunteers by gradient centrifuging. The adherent mononuclear cells were cultured. The mesenchymal stem cells(MSCs) were isolated and collected by magnetic activated cell sorting(MACS) through depletion selection by the use of CD45 and GlyA microbeads. (2) The hMAPCs were induced to differentiate with HGF(20ng/ml) +FGF-4(10ng/ml). L-02 human hepatocytes (cell lines) were used as the control, with undifferentiated hMAPCs serving as the contrast. (3)The expression of albumin (ALB), alpha fetoprotein (AFP), cytokeratin-18(CK-18), cytokeratin-19(CK-19) was detected with immunocytochemistry to identify the characters of the differentiated cells at different time, and to determine the ratio of positive cells. (4) ALB expression of the differentiated cells at different time was detected by Western blot on day 21 and 25. Results (1) The result of immunocytochemistry: The differentiated hMAPCs expressed the late markers (ALB, CK-18) of hepatocyte at all the time but did not express CK-19. The differentiated hMAPCs expressed AFP on day 7. (2) The result of Western blot assay: the differentiated cells expressed the late markers (ALB) of hepatocyte on day 21 and 35. Conclusions hMAPCs can differentiate into hepatocyte-like cells under specific inducing conditions.

Objective To observe the changes of type 2 diabetes mellitus patients in blood sugar,TC,TG,HDL C,plasma insulin,body mass index(BMI), microalbuminuria,etc before and after moderate intensity exercise training.Methods 28 patients with mean age of 59.0?7.6 years did moderate intensity exercise training for 1～1.5 hours every day for 12 month.At the same time they received the treatment of diet and oral antidiabetic drug com pared with 40 controls without exercise training. Before and after the observing period we measured the values of items above.Results Plasma glucose level , HbAlc,AUCGS(the area under the curve of glucose sugar) and BMI in fat diabetes with training team were obviously decreased compared with those before observation( P 0.05).Conclusion Exercise training can significantly descend blood sugar, insulin resistance and BMI of obese people ,improve lipidmetabolism and ascend insulin sensitivity. So the action of exercise can't be replaced by the diet and oral drug.

Accidents, Occupational ; Humans ; Hydrogen Sulfide ; poisoning

Adult ; Embolism, Amniotic Fluid ; etiology ; therapy ; Female ; Humans ; Pregnancy

Objective To evaluate the security of big animals with a new bioartificial liver system .Methods Six Tibet pigs re-spectively received treatment of hybrid artificial liver and simple bioartificial liver ,observed and recorded the vital signs ,venous pressure ,transmembrane pressure and slurry pot pressure each hour ,and collected blood to make endotoxin and bacterial culture test in the zero hour ,fourth and eighth hour .Results Compared with the zero hour ,venous pressure ,transmembrane pressure ,slur-ry pot pressure were much higher in the fifth hour (P< 0 .05) ,and there were no significant difference in the rest of other time points(P>0 .05) .The mean arterial pressure and respiratory rate in all time point showed no significant changes (P>0 .05) .Com-pared with the zero hour ,the heart rate was much lower in the second hour (P<0 .01) .The values of blood endotoxin were less than 0 .5 EU/mL in the zero hour ,fourth and eighth hour from beginning ,and the bacterial culture test showed no growth of bacteria . Conclusion The experiment of big animals with a new bioartificial liver system was safe ,the efficacy in the treatment of hepatic failure could be assessed further .

Background Heat shock proteins (HSPs) are highly conserved proteins that are induced in cells when confronted with a wide variety of proteotoxic stresses.HSP27 has a high degree of similarity with α-crystallin protein.The abnormality of HSP27 structure and expression are closely related to the formation of cataracts.Our previous study showed sodium salieylate has the protective effect on H2O2-induced lens damage.Objective This study was to investigate the roles of MAPK signal pathway in sodium salicylate-induced the expression of HSP27 in human lens epithelial cells (LECs) in vitro.Methods Human LECs were incubated in the fresh media containing sodium salicylate at different concentrations (0-55 mmol/L) for different times (1-5 hours) and allowed to be recovered in fresh medium without sodium salicylate for 1-24 hours with or without pretreatment with P38MAPK inhibitor (SB203580), ERK1/2 inhibitor (PD98059) and JNK/SAPK inhibitor ( SP600125). The expressions of P38MAPK, EBK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 were detected by Western blot. HSP27 mRNA was detected by RT-PCR. The expression of HSP27 was also detected by immunohistochemistry. Results There was only weak expression of HSP27 in normal human LECs.After stimulation of 35-55 mmol/L sodium salicylate was removed and human LECs were cultured again for 6 hours,the expression of HSP27 in LECs were significantly increased ( F= 509. 953,P<0. 01). HSP27 was absent expressed in human LECs in 55 mmol/L sodium salicylate stimulation for 1-5 hours groups, but LECs were re-cultured for 3,6 hours after removed the stimulation, the expression of HSP27 was elevated (F = 452. 534, P<0. 01). Activation of P38 M APK occurred after sodium salicylate stimulation 30 minutes and 1 hour ( F = 865.68, P<0. 01). However, ERK 1/2 was expressed after sodium salicylate was eliminated for 1-6 hours ( F = 388.84, P<0. 01). JNK/SAPK was inactived by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059. Conclusion Sodium salicylatc can induce the expression of HSP27 in human (LECs) . The effects are mediated,at least in part ,through the activation of P38MAPK and ERK1/2 signaling pathway .

Objective To retrospectively study the quantitative features and diagnostic value of 3-dimensional shear wave elastography (3D-SWE) in breast lesions.Methods A total of 198 consecutive women with 198 breast lesions (125 malignant,73 benign) were included,who underwent conventional ultrasound (US) and 3D-SWE before surgical excision.Quantitative parameters of transverse planes,sagittal planes and coronal planes were calculated,including maximum elasticity (Emax-w),mean elasticity (Emean-w),standard deviation (Esd) of the whole lesion and ratio between the stiffest elasticity (Emean-s)in the lesion and the fatty tissue (Eratio).Area under ROC curve(AUC) for combination of quantitative parameters and US were calculated.Results The AUC,sensitivity and specificity for US were 0.919,88.0％ and 78.1 ％,respectively.In the total 198 lesions,Emax-w,Emean-w,Esd,Eratio and Emean s were significantly lower in coronal planes than those in transverse and sagittal planes (all P ＜0.001).AUC for combination of each quantitative parameter and US were significantly higher than those of US (all P ＜0.05),except Emean-w of transverse plane,while there was no significant difference among the three orthogonal plane (P＞0.05).Sensitivity significantly increased by combining US with Emean-s,Eratio (transverse,sagittal and coronal planes),Emean-w (coronal and sagittal planes) or Esd (coronal and transverse planes) (all P＜0.05).Combination of US and Emean w of coronal plane yielded significantly higher sensitivity than those of transverse and sagittal planes.Conclusions Combination of quantitative features of 3D-SWE and US can significantly increase diagnostic accuracy and sensitivity in breast lesions.Emean-w of coronal plane yields the highest sensitivity.

Objective To investigate the expression of extraeellular signal-regulated protein kinase (ERK1/2)pathway in rat brains with fluorosis and the effects of fluoride on learning and memory of the rats,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of the ion.Methods Seventy-two SD rats were divided into 3 groups and 24 rats were in each group.Three groups were fed respectively with different concentrations of fluoride(NaF)for 6 months to establish rat models with fluorosis.Controls were fed with tap water (NaF＜0.5 mg/L):lower and higher concentration group were fed with water containing NaF(5,50 ms/L).Animals are sacrificed after 6 months of treatment with fluoride and the dissected brains were kept for analysis.The protein levels of ERK1/2 in rat brains were detected by Western-blotting and the mRNA level by RT-PCR. The spatial learning and memorizing ability was measured by Morris water maze test. Results The ERK1/2 protein in control group,lower and higher concentration group was 0.944±0.10,1.253±0.02,1.953±0.07,the differece being statistieally sighificant between any two groups (P ＜ 0.05). The phospho-ERKl/2 protein in control group,lower and higher concentration group was 0.73±0.08,0.77±0.07,1.28±0.11,the differece being statistieally sighificant between any two groups(P ＜ 0.05);the activation rate of phospho-ERK1/2 in lower and higher concentration group [(68.4± 3.8)%,(64.1±3.2)%] was decreased compared to control group[ (82.3±10.7)%],the differece being significant(P ＜ 0.05). In the navigation trial,longer escape latencies of lower concentration group on the second, the third,the fifth and the sixth day were observed[ (46.0±8.0),(24.0±2.7),(8.9±5.3),(7.4±4.1 )s] compared to the control[ (39.3±6.9),(19.1±9.1 ),(8.3±3.4),(4.8±2.7)s],the differece being significant (P ＜ 0.05 or ＜ 0.01 );the similar results were also observed in the higher concentration group[ (36.9±16.8),(37.7±12.9), (19.7±7.6),(12.2±5.7 )s],and the escape latencies of the higher concentration group on the third,the fifth and the sixth day were longer than that in lower concentration group. In the probe test,the rats took more time to reach the first cross in lower and higher concentration group[(1.17±0.75),(4.18±1.10)s] than control group[ (5.89± 0.56 ) s ],the differece being significant (P ＜ 0.05 or ＜ 0.01 ) ;stayed shorter [ ( 17.05±4.25 ),(18.20±4.57 ) s ] than control [(25.37±5.65 )s ] in platform area (P ＜ 0.01 );the activation rates of ERK1/2 were directly correlated with the time taken to reach the first cross platform located in the probe test(r = 0.364,P ＜ 0.05) and the activation rates were also directly correlated with the escape latencies on the sixth day(r = 0.497,P ＜ 0.05). Conclusion Long-term exposure of excessive fluoride induces the change of expression and activating rate of the ERK1/2 in rat brains,leading to the decreased capacity of learning and memory.

Objective To prepare a innovative VP3 gene‐loaded ultrasound microbubble decorated with TATp and SDF‐1 , having the extracellular accumulation and intracellular permeation function ,and characterize their property .Methods VP3 gene‐loaded ultrasound microbubbles were prepared with the method of W/O/W double emulsion .SDF‐1 and TATp were covalently con‐gjugated to the surface of poly‐lactic/acid‐glycolic acid(PLGA) microbubble though thioether bonds to prepare gene‐loaded targeted ultrasound mirobubbles .Their particle size ,distribution and surface potential were determined by Malvern measuring instrument . The conjugation status of TATp and SDF‐1 were evaluated by flow cytometry and confocal microscopy .Their DNA protection were identified by digestion reaction test .The vitro targeting capacity was preliminarily assessed by light microscopy and flow cytometry , and the vitro ultrasound imaging was investigated under high frequency imaging condition .Results The gene‐loaded targeted ultra‐sound mirobubbles showed regularly sphericity .The diameter was (536 .00 ± 16 .55)nm ,and showed a narrow distribution .The zeta potential was(-0 .08 ± 0 .08)mV .The average gene loading was 0 .62% ,with the average rate of 36 .13% gene encapsulation effi‐ciency .The DNA protective effect sustained 60 min without damage .Connection rate of TATp and SDF‐1 coupled with PLGA mi‐crobubbles surface was 69 .84% .The vitro targeting study showed that more targeted microbubbles stably clustered together in the tongue SCC‐15 cell membranes with the connection rate of 91 .44% ,while non‐targeted microbubbles combination rate was 12 .96% .Moreover ,the vitro ultrasound imaging was tiny dot ,even high echo .Conclusion TATp‐SDF‐1‐VP3‐PEG‐PLGA micro‐bubbles were prepared successfully ,which can efficiently target to tongue SCC‐15 cells ,and pass through the cell membranes at a short time in company with outstanding ultrasound imagings in vitro .

Objective To retrospectively study the anisotropy of shear wave elastography (SWE)quantitative parameters of breast lesions,and the correlation with histopathology.Methods A total of 281 consecutive women with 281 breast lesions (179 malignant,102 benign) were included,who underwent conventional ultrasound (US) and 2D SWE before surgical excision.Three acquisitions each for transverse and longitudinal planes were obtained,and maximum elasticity (Emax),mean elasticity (Emean),standard deviation (Esd) of the whole lesion and ratio between the elasticity in the mass and the fatty tissue (Eratio)were recorded.Anisotropic difference (AD) and anisotropy factors (AF) were calculated,and correlation with histopathology was analyzed.Results The average Emax,Emean and Esd of transverse planes were significantly higher than those of longitudinal planes.AF showed positive correlation with quantitative elasticity (Emax,Emean,Esd and Eratio) (P =0.000),and was significantly higher in malignant lesions than that in benign besions (P =0.000).AUC of AF was significantly higher than that of AD (P ＜0.001).AF was significantly higher in invasive ductal carcinoma than that in ductal carcinoma in situ.Higher AF was associated with higher histopathological grades of invasive ductal carcinoma (P =0.000),and correlated with ER/PR(+).Conclusions Anisotropy of SWE is an indicator of malignancy of breast lesions,and is of predictive value for prognosis in breast cancer.