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.

Objective:To study Clinical Effect of Anterolateral Approach ofor the Patients with Hip Arthroplasty and the effect on the serum CRP,IL-6,D-Dimer in levels.Methods:102 Patients with Hip Arthroplasty who received therapy from February 2013 to January 2015 in our hospital,Patients were asked to be divided into observation group and control group.Among them,the control group of 54 cases,through the conventional approach to hip arthroplasty;48 cases of observation group.Serum CRP,IL-6 and D-dimer levels were compared between the two groups before and after surgery,and the efficacy of the two groups was compared by follow-up.Results:After treatment,the total effective rate of the observation group was significantly higher than that of the control group (P＜0.05),Serum CRP,IL-6 and D-dimers increased in both groups at 12 h after surgery and decreased gradually at 7 and 14 days.The levels of serum CRP,IL-6 and D-dimer in the observation group were significantly lower than those in the control group at 12 h,7 d and 14d(P＜0.05).Conclusion:The clinical effect of the anterolateral approach on hip arthroplasty is more significant than that of conventional approach,and can effectively reduce the levels of serum CRP,IL-6 and D-dimer,and is more favorable for postoperative recovery.

Objective To explore the change and the spectra of opportunistic infections in AIDS patients after antiretroviral treatment in He′nan province.Methods Inpatients of post treatment with disease in accordance with the criteria were selected from the Sixth People′s Hospital of Zhengzhou.A retrospective study was carried out, including the content such as gender,age,infection route,treatment time,the date of complication,and CD4+T lympho-cyte counts when the patient with complications.Disease were divided into four group:opportunistic infections,adverse drug reactions,tumor,cardio-cerebrovascular disease and diabetes mellitus.Statistic case number and incidence of every kinds of the disease in the different stages of HAART.Results (1)1 001 cases of patients were involved in the result analysis.People of 41-60 years old,the peasants and peasant workers,accounted for 57.4%and 78.9%.The infection route was mainly through blood transmission and blood collection, accounting for 83.45%.Most of the patients come from Zhumadian, Xuchang, Zhoukou, Nanyang, which were high -incidence area in He′nan.( 2 ) In 1 001 patients of post treatment, the rate of opportunistic infections was 58.4%, while adverse drug reactions was 30.5%.Bacterial pneumonia,extrapulmonary tuberculosis,oral cavity fungal infection and tuberculosis were common in opportunistic infections of patients of post treatment.The main adverse reactions were allergic dermatitis,liver injury and myelosuppression.The liver cancer case was the highest ( 8/29 ) .( 3 ) 1 001 cases were divided into 6 groups according to different count of treatment time,during 3 months of treatment,the incidence rate of opportunistic infec-tions and adverse drug reactions was the highest,accounted for 75.2% and 41.6%.After 5 years of treatment,the incidence rate of cardio-cerebrovascular disease and diabetes mellitus was 6.39%.With the prolonging of HAART time,incidence of opportunistic infections and adverse drug reactions remarkably reduced,but the occurence rate car-dio-cerebrovascular disease and diabetes mellitus increased.Conclusion In AIDS patients after HAART in He′nan at present,opportunistic infections and adverse drug reactions are main diseases and used to occur during three months after HAART.Attention will be focused on tumor, cardio -cerebrovascular disease and diabetes mellitus in AIDS patients of long-term survival.

Objective To study the solute clearance effect of the new concentrated anticoagulation hemodiaiysate of citrate for hemodialysis in patients with high risk of bleeding. Methods Forty-two kidney failure patients with high risk of bleeding were divided into two groups (Group A and Group B) according to their hemodialysis manners. Patients in Group A were hemodialyzed with bicarbonate hemodialysate with low-molecular-weight heparin (dalteparin) anticoognlation and those in Group B with the new citrate antieoagnlation hemodialysate prepared in our hospital without any other anticoagulant. Blood urea nitrogen (BUN) and creatinine (Cr) concentrations were measured before and after dialysis, and Kt/V and urea reduction rate (URR) were calculated. In addition, activated clotting time (ACT) and ionized calcium (iCa2+) concentration were also measured at the arterial and venous ends. Results ACT was extended and iCa2+ concentration decreased significantly at the venous end compared with those at the arterial end in Group B (P<0.01). BUN and Cr concentrations were markedly decreased after dialysis compared with those before dialysis in both groups (P<0.01), and no significant difference in solute clearance effect, as indicated by Kt/V and URR, was observed between Group A and Group B (P>0.05). Conclusion The solute clearance effect of the new concentrated anticoagnlation hemodialysate of citrate is excellent during hemodiaiysis in kidney failure patients with high risk of bleeding.

Objective To analyze the frail status and influencing factors of inpatients in geriatrics department.Methods A total of 170 patients aged ≥ 65 years in Geriatrics Department in our hospital were selected.Frailty was defined according to the Clinical Frailty Scale (CFS),and patients were divided into non-frail group and frail group.The differences in comorbidity,physical function,nutrition,cognitive,psychological,geriatrics syndromes,medication,social support and other aspects were analyzed between the two groups by using Comprehensive Geriatrics Assessment(CGA),and correlationof the factors with frailty was evaluated.Results There were 83 (49.0％)patients in frail group and 87 (51.0％)in non-frail group.In frail group versus non-frail group,Charlson comorbidity index was(2.7±2.0)vs.(1.9±1.1),and cumulative illness rating scale was(10.8±3.7) vs.(8.0±2.6) (all P＜0.05);mini-nutrition assessment was(11.2±2.3)vs.(13.0±1.3),and simple cognitive score was(2.7±1.3)vs.(4.1±0.9)(all P＜0.05).Grip,five chair rising,timed get-up and go test,walking speed,SPPB showed statistically significant differences (all P＜ 0.05) between frail group and non-frail group:(20.7±6.6)kg vs.(27.96.7)kg,(30.4±17.5)vs.(12.9±4.1),(23.7± 9.9)vs.(11.7± 3.3),(0.5 ± 0.2)m/s vs.(0.9± 0.2)m/s,(5.6 ± 2.3) vs.(10.3±1.8)respectively.The incidences of incontinence,visual impairment,hearing impairment,sleeping disorder,oral problems,chronic pain,anxiety or depression and fall history were higher in frail patients than in non-frail patients.Living alone,the widowed,nursing staff employment proportion were higher in frail group than in non-frail group.Level of education was lower in frail group than non-frail group.Logistic regression analysis showed that BMI,the walking speed and the number of geriatrics syndromes had a significant impact on frailty(OR=0.789,0.000,2.745,all P＜0.05).Conclusions Incidence rate of frailty in elderly hospitalized patients is high.To identify frailty of hospitalized patients,comprehensive geriatric assessment can be used for understanding frail characteristics and its influencing factors for hospitalized old patients,so as to provide evidence for the reasonable treatment programs.

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.

Objective To investigate the effect of MAPK activation on autophagy in the hippocampus of rats with subarachnoid hemorrhage.Methods A total of 100 male SD rats were divided into 5 groups randomly:sham operated group,SAH group,inhibitor U0126 group,inhibitor SB203580 group,SP600125 group.The animal model was established by injecting the autologous blood into cisterna magna twice.The morphological changes of hippocampus nerve cells of rat brain were detected with HE.The mRNA levels of ERK1/2,p38MAPK,JNK and LC3 in hippocampus were detected with quantitative real time PCR and the expression of phosphorylated ERK1/2,phosphorylated p38MAPK,phosphorylated JNK and LC3-Ⅱ in hippocampus of rat brain were detected by immunohistochemistry.Results Compared with the Sham group,the survival rate of neurons in SAH group decreased (6 h:(84.982 ± 5.723) ％,24 h:(74.383± 9.860) ％,48 h:(62.860± 10.820) ％,72 h:(52.260± 10.960) ％) (all P＜0.05).The levels of ERK1/2 mRNA,p38MAPK mRNA,JNK mRNA and LC3 mRNA in hippocampus increased (all P＜ 0.05) and the expression of p-ERK1/2,p-p38MAPK,p-JNK,LC3-Ⅱ proteins increased(all P＜0.05).Compared with the SAH group,the survival rate of neurons in U0126 group was decreased (6 h:(71.620±6.542) ％,24 h:(66.221±7.742)％,48 h:(55.208±8.802) ％,72 h:(46.242±7.782) ％),and the ERK1/2 and LC3 in hippocampus decreased both in mRNA level and in protein level(all P＜0.05).Compared with the SAH group,the survival rate of neurons in SB203580 groups was increased (6 h:(89.082±6.602)％,24 h:(85.840±9.726) ％,48 h:(74.96± 10.916) ％,72 h:(69.211 ± 10.745) ％),and the p38MAPK,LC3-Ⅱ in hippocampus decreased at both mRNA and protein levels (all P＜0.05).Compared with the SAH group,the survival rate of neurons in SP600125 groups was increased (6 h:(91.620± 7.542) ％,24 h:(86.221 ± 10.742) ％,48 h:(75.208±11.802) ％,72 h:(70.242± 11.782) ％).The expression of JNK was decreased while the LC3-Ⅱ was increased in hippocampus (P＜0.05).Conclusion MAPK activation is involved in the autophagy of hippocampal neurons after SAH,in which ERK1/2 activation plays a positive role in the regulation of autophagy in hippocampal neurons after SAH,while p38MAPK and JNK activation plays a negative role in autophagy.

Adult ; Bone Cysts, Aneurysmal ; diagnosis ; surgery ; Bone Neoplasms ; diagnosis ; surgery ; Chondroblastoma ; diagnosis ; surgery ; Female ; Humans ; Male ; Talus ; surgery ; Young Adult

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

The stems and branches of Hypericum petiolulatum were extracted by alcohol and liquid-liquid extraction. Seven furofuran lignans were isolated from the ethyl acetate fraction of ethanol extract of H. petiolulatum by using silica gelchromatography, Sephadex LH-20 chromatography, medium-pressure liquid chromatography and preparative HPLC. Their structures were identified by the spectroscopic methods as pinoresinol (1), medioresinol (2), 8-acetoxypinoresinol (3), epipinoresinol (4), (+)-syringaresinol (5), (+)-1-hydroxysyringaresinol (6) and erythro-buddlenolE (7). All the isolates were firstly found in H. petiolulatum. In the bioassay, compound 7 showed remarkable antioxidative activity inhibiting Fe(+2)-cystine induced rat liver microsomal lipid peroxidation with inhibitory rate 38% at a concentration of 1 x 10(-6) mol · L(-1) (positive control Vit E with the inhibitory rate of 35% at the same concentration).
Animals ; Antioxidants ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Hypericum ; chemistry ; Lipid Peroxidation ; drug effects ; Microsomes, Liver ; drug effects ; metabolism ; Molecular Structure ; Oxidative Stress ; drug effects ; Plant Stems ; chemistry ; Rats