Objective To investigate the effect of human hepatocellular carcinoma HepG2 cell-derived Exosome on the differentiation of mesenchymal stem cells(MSC)into cancer-associated myofibroblasts(CAF)and the impacts of CAF on liver cancer cell proliferation,migration,and invasion. Methods The protein expression of HepG2 cell-derived Exosome was detected by Western blotting. MSCs were separated from human adipose tissue and cultured with HepG2 cell-derived Exosome(100 ng/nl)to initiate differentiation. The expressions of mesenchymal markers and several interleukins were also detected by Western blotting. HepG2 cells were co-cultured with the conditioned media(CM),in which HepG2 Exosome induced the differentiation of MSC into CAF. The expressions of epithelial and mesenchymal markers were detected by real-time polymerase chain reaction(PCR)and Western blotting. Cell proliferation was assessed using MTS assay. Transwell chambers were used in the in vitro migration and invasion assay. Results HepG2 cell-derived particles expressed CD63,70 kilodalton heat shock proteins,and 90 kilodalton heat shock proteins. With the treatment of HepG2 cell-derived Exosome,the expressions of mesenchymal marker α-smooth muscle actin,fibroblast activation protein α,interleukin(IL)-6,IL-8,and IL-1β were up-regulated,while vascular endothelial growth factor had no significant change. The conditioned media which HepG2 Exosome induced MSC differentiation CAF(CAF-CM)could significantly promote HepG2 cells proliferation(1.075±0.104),compared to BSA control(0.874±0.066,P=0.023)and MSC-CM(0.649±0.034,P=0.0005). CAF-CM could significantly enhance cell migration [(42.5±9.1) cells vs.(18.5±3.1) cells,P=0.001] and invasion [(29.0±3.5) cells vs.(13.1±3.7) cells,P=0.009] compared to its control group. Moreover the conditioned medium which HepG2 Exosome induced MSC to differentiate into CAF could also promote the expressions of mesenchyme-related genes Smad interacting protein 1(P=0.040),β-catenin(P=0.038),fibronectin(P=0.029),and Vimentin(P=0.013)and inhibit the expression of epithelial related genes zonula ocdudens-1(P=0.010).Conclusions Exosome extracted from HepG2 cells can induce human adipose-derived MSC to differentiate into cancer-associated myofibroblasts. CAF-like cells can promote the migration of the liver cancer cell line HepG2.

T_4 was coupled to human serum albumin(HSA)to make the conjugate T_4-HSA,a polyvalent T_4 analogue rather than monovalent T_4 per se alone.A new strategy for sandwich enzyme immunoassay model system of small molecular hapten hormones FT_4 was developed,such as intra-assay CV,inter-assay CV,average recovery, normal and abnormal values were fit for clinical application.The sensitivity was as much as 10 times higher than conventional competitive enzyme immunoassay.

Objective To explore the changes and significances of serum leptin and interleukin-1?(IL-1?)before and after treatment with risperidone in children with schizophrenics.Methods Thirty-one cases with first-episode children with schizophrenia were chosen in patient group.Their body weight and height were measured before and 8 weeks after treatment with risperidone to calculate the body mass index(BMI),and their fasting serum leptin and IL-1? were assayed radio-immunity method.Thirty-one healthy controls were measured at the same time.Results BMI and the level of serum leptin in patients of post-treatment increased significantly compared with those of pre-treatment(all P

Objective To analyze the clinical and radiological features of retroperitoneal fibrosis (RPF), and to deepen the understanding of this unusual disease and improve the diagnostic level at the early stage. Methods Fourteen cases (10 males and 4 females, mean age 45.8 years) of pathologically diagnosed RPF from January 1990 to June 2004 were summarized. The clinical and radiological performance of the cases were analyzed. All patients received non-contrast CT scanning, 10 of them underwent enhanced CT scanning as well. 8 patients received MRI, 10 patients received IVP examination, and 11 received B-ultrasound. Results (1)The very first symptoms usually included back pain, bellyache (10 cases), or urinary tract obstruction (3 cases), with increase of ESR, IgG, CRP value and abnormal renal function.(2)The result of radiological examination showed that 11 lesions of the 14 cases located at retroperitoneum. Ten cases were mass type and 4 cases were diffuse type. Non-contrast CT scanning revealed soft tissue mass at retroperitoneum with inhomogenous or homogenous density. After contrast medium injection the lesions were enhanced with different extent. MRI results showed that the lesions presented low signal in T_1WI, while in T_2WI the signals had no obvious coherence but were different from one case to another. Conclusion Radiological examination is one of the important methods for diagnosis of RPF. Based on the different characteristics of RPF in CT and MRI, together with the clinical findings, we will get valuable references for staging and follow-up of RPF.

Objective: To observe the effect of different moxibustion durations on rats with rheumatoid arthritis (RA) and to evaluate the relationship between moxibustion amount and moxibustion efficacy.Methods: Eight rats were randomly selected as a normal group from the 40 male Sprague-Dawley (SD) rats, and the other 32 rats were used to establish typeⅡ collagen-induced RA models. After successful modeling, the 32 rats were randomly divided into a model group, a moxibustion for 20 min group, a moxibustion for 40 min group and a moxibustion for 60 min group, with 8 rats in each group. Rats in the normal group did not receive modeling and moxibustion intervention; rats in the model group did not receive moxibustion after modeling; rats in the moxibustion for 20 min group, the moxibustion for 40 min group and the moxibustion for 60 min group were treated with moxibustion at Shenshu (BL 23) and Zusanli (ST 36) for 20 min, 40 min and 60 min, respectively. Six days were a course of treatment, with a total of 3-course treatments and a 1-day rest between the courses of treatment. After treatment, the serum levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α, arthritis index (AI) scores, toe volumes and pathological score of synovitis were evaluated in the rats.Results: Compared with the normal group, the serum IL-1β and TNF-α levels, and the toe volumes in the model group were increased, and the differences were statistically significant (P<0.01), before the treatment. Compared with the model group, the serum IL-1β and TNF-α levels, toe volumes and arthritis index (AI) scores were significantly decreased in the moxibustion for 20 min group, the moxibustion for 40 min group and the moxibustion for 60 min group (P<0.05 orP<0.01 ). Compared with the moxibustion for 20 min group and the moxibustion for 60 min group, serum IL-1β and TNF-α levels, toe volumes and AI scores were decreased more significantly in moxibustion for 40 min group, and the differences were statistically significant (P<0.05 orP<0.01). There were no significant differences in serum IL-1β and TNF-α levels, AI scores and toe volumes between the moxibustion for 20 min group and the moxibustion for 60 min group (allP>0.05). The synovial histopathological improvement was the most obvious in the moxibustion for 40 min group, when the synovial histopathological changes were compared among the moxibustion for 20 min group, moxibustion for 40 min group and moxibustion for 60 min group.Conclusion: The therapeutic efficacy of moxibustion for 40 min in RA rats was more significant than that of moxibustion for 20 min and moxibustion for 60 min, indicating that the duration of moxibustion is the main factor affecting its therapeutic efficacy.

OBJECTIVETo investigate the effects of different concentrations of lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), dexamethasone (Dex), and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro.

METHODSMCF7 breasr cancer cells were cultured and treated with different concentrations of LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L). Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot, respectively, after 6, 24, 48, and 72 hours of treatment.

RESULTSCompared with the blank con- trol, LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L) significantly increased the expressions of GPR54 mRNA (P < 0.05) and protein (P < 0.05).

CONCLUSIONLPS, TNFα, IL-6, Dex, and insulin evidently increase the expression of GPR54 in the MCF7 cell line, indicating their influence on the function of gonads by regulating the GPR54 level.

Blotting, Western ; Dexamethasone ; administration & dosage ; pharmacology ; Glucocorticoids ; administration & dosage ; pharmacology ; Gonads ; drug effects ; metabolism ; Humans ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Interleukin-6 ; administration & dosage ; pharmacology ; Lipopolysaccharides ; administration & dosage ; pharmacology ; MCF-7 Cells ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, G-Protein-Coupled ; drug effects ; genetics ; metabolism ; Receptors, Kisspeptin-1 ; Time Factors ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology

Objective To investigate the relation between lesions of medullary infarction and clinical symptoms and signs.Methods Eleven patients with medullary infarction confirmed by MRI were identified.Their clinical presentations and the relation between clinical presentation and location of lesion in MRI were assessed and analyzed.Results Among the 11 patients,5 cases suffered from lateral medullary infarction,6 medial medullary infarction(including two cases of bilateral infarction).Regarding clinical symptoms,common symptoms in lateral medullary infarction are blurred speech,dizziness,choking water,swallowing difficulties and facial numbness;in medial medullary infarction are limb weakness and blurred speech.The common signs of lateral medullary infarction are dysarthria,sensory disturbance,Horner sign and facial paralysis;in medial medullary infarction are paralysis of limbs and peripheral paralysis of the tongue.Conclusion Clinical manifestations are vailed depending on the sites of medullary infarction.MRI is helpful in diagnosing of medullary infarction.

OBJECTIVETo explore the impact and mechanism of electro-acupuncture (EA) on olfactory ensheathing cells (OECs) transplantation of spinal cord injury (SCI) axonal regeneration.

METHODSIn the experiment, 72 adult Sprague Dawley male rats weighted (220±20) g underwent contusion and transection method to cause the T9 model of spinal cord injury, were randomly divided into four groups involving model group, EA group,OECs group,and EA+OECs group. 5% fluorescein gold (FG) solution of 0.5 µl was injected into rats' spinal cord at 4 weeks and 8 weeks after SCI, a series of tests were performed including fluorescein gold(FG) retrograde tagging, BBB scores.

RESULTS(1)The BBB scores level among four groups had no differences from the 1st day to the 1st week after the SCI (P>0.05). From the 3rd week after the SCI, the BBB scores level in EA+ OECs group were obviously higher than that of other groups (P<0.05). (2)The result of the fluorescein gold (FG) retrograde tagging showed at 4 weeks and 8 weeks after treatment FG positive nerve fibers were observed in SCI region. In EA+OECs group the number of FG positive nerve fibers was more than other three groups, and the fibers were more regularly arranged than other three groups.

CONCLUSIONThe combination of electro-acupuncture and OECs transplantation can recover the pathway of nerve conduction and promote nerve fibers regeneration and hind limb function recovery for SCI rat, and can guide the trend of the axonal regeneration.

Animals ; Axons ; physiology ; Cell Transplantation ; Combined Modality Therapy ; Electroacupuncture ; Humans ; Male ; Nerve Regeneration ; Olfactory Nerve ; physiopathology ; transplantation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; physiopathology ; therapy

Objective To investigate the diagnostic significance of endoscopic findings in Henoch-Schonlein purpura(HSP),especially when abdominal pain preceded the cutaneous lesions.Methods The clinical data and gastroscopic findings in 37 cases of children with HSP were studied and analysed retrospectively in order to detect the pathological changes in the stomach and duodenum mucosa.The biopsy was taken in the pathological changeing place,and the relationship between clinical and endoscopic findings was analyzed.Results Detection rate of the pathological changes in the stomach and duodenum mucosa was 62.2%,31.3% of which experienced only cutaneous lesions,100% of which presented the acute abdominal pain.Three patients were not checked up the pathological changes.Of them,1 had arthritis,2 had Henoch-Schonlein nephritis.Characteristically endoscopic findings in the stomach and duodenum mucosa were found.The endoscopic findings included anabrosis,hyperemia,edema and hemorrhage.Conclusions Detection rate of the pathological changes in the stomach and duodenum mucosa is higher.Endoscopy is very helpful to the early diagnosis of HSP in children,especially abdominal pain presented firstly.

Objective To evaluate the effect of propofol on mitochondrial fission in a rat hippocampal neuron model of hypoxia/reoxygenation (H/R) injury.Methods Primarily cultured hippocampal neurons of neonatal Sprague-Dawley rats were randomly divided into 4 groups (n =42 each) using a random number table:control group (group C);vehicle group (group V);H/R group;H/R+propofol group (group H/R+P).In group V,H/R was not produced,the vehicle dimethyl sulfoxide with the final concentration of 0.01％ was added,and the cells were then incubated for 6 h.In group H/R,the hippocampal neurons were subjected to oxygen-glucose deprivation for 6 h followed by 20 h reoxygenation.In group H/R+P,propofol with the final concentration of 1 μmol/L was added at 6 h of hypoxia.At 20 h of reoxygenation,the cell apoptosis (using flow cytometry),Ca2+ concentrations in cytoplasm (with the laser scanning confocal microscope),calcineurin (CaN) activities (by enzyme-linked immunosorbent assay),and expression of mitochondrial fission proteins dynamin-related protein 1 (Drp1) and fission 1 (Fis1),and apoptosis-related proteins cytochrome c (Cyt c) and apoptosis-inducing factor (AIF) (by Western blot) were measured.The apoptotic rate was calculated.Results Compared with group C,the apoptotic rate,Ca2+concentrations,CaN activities,and expression of Drp1,Fis1,Cyt c and AIF were significantly increased in H/R and H/R+P groups (P＜0.05),and no significant changes were found in the parameters mentioned a2+.bove in group V (P＞0.05).Compared with group H/R,the apoptotic rate,Ca+ concentrations,CaN activities,and expression of Drp1,Fis1,Cyt c and AIF were significantly decreased in group H/R+P (P＜0.05).Conclusion Propofol can reduce the H/R injury to rat hippocampal neurons through inhibiting mitochondrial fission.