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 analyze the key factors influencing the number of diseases or conditions in which clinical pathways ( CPs) were implemented at public hospitals of China. Methods Based on the questionnaire survey of 51 public hospitals that had implemented CPs in Shanghai, Hubei province, and Gansu province, a multivariate logistic model was used to analyze the factors that influenced the number of CPs implemented in hospitals. Results In terms of the 14 issues and difficulties found in CPs′implementation, the overall acceptance rate was 38. 8% on average among surveyed hospitals, yet with a great variation(from 7. 1% to 100. 0%). A multivariate logistic model showed that the recognition of public hospitals on the issues and difficulties in CPs′implementation did not affect the number of implemented CPs, and tertiary public hospitals had significantly higher number of CPs implemented than secondary hospitals. Conclusions The resources and management capabilities of public hospitals in China are the determinants of CPs′promotion.

OBJECTIVETo explore the clinical efficacy and action mechanism of acupuncture for premature ovarian failure (POF).

METHODSAccording to prospective cohort study design, 30 cases were included. Based on theory of regulating Chong and Conception Vessels as well as soothing the liver and calming the nerves, acupuncture at Guanyuan (CV 4), Dahe (KI 12), Taixi (KI 3), Taichong (LR 3), Baihui (GV 20) was applied, three times per week and 3 months were considered as one session. Totally two sessions were performed. The menstruation condition, estradiol (E2), follicle-stimulating hormone (FSH) and scores of the clinical perimenopausal symptoms were taken as treatment outcomes.

RESULTSThe total effective rate was 86.7% (26/30) and the cured rate was 16.7% (5/30). The recovery rate of menstruation was 16.7% (5/30) and the regain rate of menstruation was 56.7% (17/30). After the treatment, the level of E2 was significantly increased from (45.41 +/- 18. 09) pmol/L to (59.07 +/- 24.21) pmol/L (P < 0.05), and the score of perimenopausal symptoms was obviously reduced from 14.28 +/- 8. 30 to 4.04 +/- 3.28 (P < 0.01). However, no statistical differences could be seen in FSH before and after treatment (P > 0.05).

CONCLUSIONAcupuncture has certain effect on improving menstruation and perimenopausal symptoms in POF patients, which is likely to he related with increasing the level of E2.

Acupuncture Therapy ; Adolescent ; Estradiol ; metabolism ; Female ; Humans ; Luteinizing Hormone ; metabolism ; Primary Ovarian Insufficiency ; metabolism ; therapy ; Prospective Studies ; Young Adult

The quality of guidelines of clinical practice in acupuncture and moxibustion was evaluated with AGREE II tool. After retrieval of officially-issued guidelines of clinical practice in acupuncture and moxibustion from 2004 to 2013, totally 5 types of evidence-based guidelines of clinical practice in acupuncture and moxibustion were acquired, including depression, herpes zoster, pseudobulbar palsy after stroke, migraine and Bell's palsy. With AGREE II tool, independent score in six areas, including guideline's scope and purpose, participants and personnel, preciseness, clarity and readability, applicability and editorial independence were analyzed to perform a comprehensive evaluation in the end. The assessment results indicated that the score of editorial independence in evidence-based guidelines of clinical practice in acupuncture and moxibustion was the highest, averagely 97.9%, which was followed by clarity and readability (83.3%), participants and personnel (78.1%), preciseness (75.6%), the scope and purpose (68.1%) and the applicability (62.5%). The total evaluation score was all 6 points in the five guidelines with recommended as comprehensive evaluation advice. This study results indicate that although the guideline of clinical practice in acupuncture and moxibustion is low in number, the total quality is considerable.
Acupuncture Therapy ; standards ; Evidence-Based Medicine ; Humans ; Moxibustion ; standards ; Practice Guidelines as Topic ; Quality Assurance, Health Care

Objective: To understand the survival status and influencing factors of HIV/AIDS patients with highly active antiretroviral therapy ( HAART ) among drug users in Yili Prefecture, Xinjiang from 2005 to 2019, so as to provide references for reducing AIDS mortality. Methods : The demographic information, clinical stage, baseline CD4+T lymphocyte ( CD4 ) level and treatment status of HIV/AIDS patients with HAART in Yili Prefecture from 2005 to 2019 were collected through AIDS Antiretroviral Therapy Information System. The survival rate was calculated by the life table method. The influencing factors for survival time were analyzed by Cox proportional hazard regression model. Results: Totally 1 935 patients were recruited, the median age receiving HAART was 37 years old and the median CD4 counts was 293/μL. The cumulative survival rates at 1, 5, 7 and 10 years were 97%, 78%, 73%, and 66%, respectively. The multivariate Cox proportional hazards regression analysis showed that the patients with body mass index of 18.5-<28.0 kg/m2 ( HR: 0.391-0.656, 95%CI: 0.234-0.958 ), baseline CD4>200/μL ( HR: 0.354-0.667, 95%CI: 0.232-0.841 ) , or missed medication in the last 7 days ( HR=0.009, 95%CI: 0.001-0.061 ) had lower risk of death; the patients with WHO clinical stage of Ⅱ-Ⅳ ( HR: 1.479-2.311, 95%CI: 1.004-3.288 ) or treatment delay ≥1 years ( HR: 1.287-1.388, 95%CI: 1.029-1.826 ) had higher risk of death. Conclusions The 5-year cumulative survival rate of HIV/AIDS patients with HAART in Yili Prefecture is 78%. Body mass index, baseline CD4 level, WHO clinical stage, treatment delay and missed medication in last 7 days were the influencing factors for survival time.

Objective:To analyze the clinical prognostic factors of liposarcoma on the extremities and trunk, as well as to retrospectively analyze the effect of adjuvant radiotherapy and chemotherapy on liposarcoma of the extremities. Methods:Patients with liposarcoma of the extremities treated in our hospital from July 1, 2007 to December 31, 2012 were followed up. The relationship of clinical prognostic factors with gender, age, location, depth, and size of the tumors, as well as the histological grade and admission status, were statistically analyzed. The effects of adjuvant radiotherapy or chemotherapy on overall survival (OS) and disease free survival (DFS) were evaluated. Results:A total of 82 patients with extremity liposarcoma received surgery-based comprehensive treatment in our hospital. Of the total patients, 73 received a 24-month to 88-month satisfied follow-up;the median follow-up time was 47 months. The OS rate was 83.6%(61/73), and the DFS rate was 68.5%(50/73). Multivariate Cox regression analysis showed that the tumor location, histological grade, and admission status were the independent correlative factors influencing DFS, and the age and pathologic grading were the independent correlative factors influencing the OS. Kaplan-Meier survival analysis showed that radiation therapy can significantly improve the DFS and OS of the G2 and G3-grade liposarcoma (DFS:59.1 months vs. 28.4 months, P<0.01;OS:70.8 months vs. 55.1 months, P<0.05). Significant difference was not found in the effect of chemotherapy on OS and DFS. Conclusion:The prognosis of liposarcoma was significantly associated with the pathologic grades and subtypes. Auxiliary radiotherapy could improve the survival and prognosis of G2 and G3 liposarcoma of the extremities, but the role of chemotherapy in treating liposarcoma remained unclear.

Objective To optimize the design of the field psychological emergency rescue chest.Methods The deficiencies of the chest were summarized during the drilling and actual application of the psychological rescue unit,and some optimization countermeasures were put forward accordingly.Results The chest was optimized from the aspects of selection of chest body, internal and external layout as well as equipped devices.Conclusion The optimized chest can better meet the needs of the disaster emergency rescue.

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

The Notch signaling pathway has been implicated in the regulation of cell-fate decisions such as differentiation of embryo stem cells and neural stem cells into neurons. We cultured human mesenchymal stem cells (hMSCs) in vitro and induced hMSCs to differentiate into neural cells by beta-mercaptoethanol (beta-ME), DMSO and 3-tert-butyl-4-hydroxyanisole (BHA). Immunocytochemistry was utilized to detect neuron-specific enolase (NSE) and Nissl body, and flow cytometry was used to determine cell growth phases. The expressions of signal molecules involved in the Notch pathway such as Notch1, Jagged 1 (JAG1), presenilin 1 (PS1) and hairy and enhancer of split 1(HES1) were observed by RT-PCR and immunofluorescent techniques. The results were as follows: (1) Before induction, the percentage of hMSCs at G(0)/G(1) was 58.5%, and the percentage at S+G(2)/M was 41.5%. After induction, the percentage of hMSCs at G(0)/G(1) increased to 73.1%, 76.2% and 78.1%, respectively on days 2, 4 and 6, and the percentage at S+G(2)/M decreased to 26.8%, 24.8% and 21.9%, respectively; The percentage of NSE-positive cells reached (77+/-0.35) %; Nisslos staining was positive in cytoplasm. (2) Notch1 and JAG1 were both expressed in hMSCs before and after induction, but the mRNA expressions of both Notch1 and JAG1, detected by RT-PCR, decreased obviously after induction(P<0.05). Notch1 mRNA/beta-actin was 1.157, 0.815, 0.756 and 0.570, and JAG1 mRNA/beta-actin was 0.437, 0.350, 0.314 and 0.362, respectively, on days 0, 2, 4 and 6 after induction. The Notch pathway activation participant PS1 mRNA and Notch pathway target gene HES1 mRNA also decreased apparently after induction (P<0.05), and their mRNA/beta-actin was 0.990, 0.449, 0.441, 0.454 and 0.370, 0.256, 0.266, 0.240 on days 0, 2, 4 and 6, respectively. These observations indicate that the expressions of Notch signal molecules were suppressed when hMSCs were induced to differentiate into neural cells. Based on these findings, we propose that low level of Notch signaling activation may contribute to neural cell differentiation.
Basic Helix-Loop-Helix Transcription Factors ; genetics ; Calcium-Binding Proteins ; genetics ; Cell Cycle ; Cell Differentiation ; Flow Cytometry ; Homeodomain Proteins ; genetics ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Jagged-1 Protein ; Membrane Proteins ; genetics ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Receptor, Notch1 ; genetics ; Receptors, Notch ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Serrate-Jagged Proteins ; Signal Transduction ; Transcription Factor HES-1

BACKGROUNDChronic heart failure (CHF) had been characterized as an activated sympathetic system leading to the alteration of adrenergic receptor (AR) levels in the heart. Thus far, not much research has been done with regard to traditional Chinese medical treatment for CHF. We investigated the effect of Shexiangbaoxin pills (SXBXP) on the function of the heart and the expression of a(1)-AR and b-AR subtypes in the messenger RNA (mRNA) levels and protein levels of non-infarction left ventricular tissue from rats with CHF induced by myocardial infarction.

METHODSModels of CHF were established by left anterior descending coronary artery ligature. Fifty-four Wistar rats were randomly divided into five groups: normal control group (group A), sham operation group (group B), CHF model group (group C), positive medicine control group (group D), and small-dose SXBXP group (group E) and large-dose SXBXP group (group F), deployed intragastrically. Cardiac function was examined by echocardiography before and after therapy; mRNA expressed levels were measured by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) for b(1)-AR, b(2)-AR, b(3)-AR, a(1A)-AR, a(1B)-AR, and a(1D)-AR; protein levels were measured by Western blotting analysis for b(1)-AR, b(2)-AR, a(1A)-AR, a(1B)-AR, and a(1D)-AR in non-infarction left ventricular tissue.

RESULTSThere was no significant difference in the left ventricular ejection fraction (LVEF) between groups A and B. Compared to group B, LVEF of groups C, D, E, and F were significantly decreased (P < 0.01) before therapy. After therapy, compared to group C, LVEF of group F was significantly improved (P < 0.05). Compared to group B, b(1)-AR and a(1B)-AR expressed levels were markedly decreased (P < 0.05), a(1A)-AR and b(3)-AR were significantly increased (P < 0.01) in group C, and in both mRNA and protein expressed levels b(2)-AR had no significant difference between groups B and C (P > 0.05). a(1D)-AR mRNA levels were unchanged in each group (P > 0.05), but a(1D)-AR protein level was significantly decreased in group C (P < 0.05). After treatment, compared to group C, mRNA levels of b(1)-AR and a(1B)-AR were significantly increased (P < 0.05 and P < 0.01), and a(1A)-AR was markedly decreased in groups D, E, and F (P < 0.05). b(3)-AR level significantly declined in both groups D and F (P < 0.01), but b(2)-AR and a(1D)-AR expressed levels remained unchanged in each group (P > 0.05). Protein levels, compared to group C, b(1)-AR was significantly increased (P < 0.01, P < 0.05, and P < 0.01) and a(1A)-AR was markedly decreased in groups D, E, and F (P < 0.05, P < 0.01, and P < 0.01). b(2)-AR expressed level was significantly increased in group F (P < 0.05). a(1B)-AR expressed level was significantly increased in both groups E and F (P < 0.05), and a(1D)-AR was remarkably increased in both groups D and F (P < 0.05).

CONCLUSIONSAfter SXBXP treatment, LVEF was increased and cardiac function was significantly ameliorated in rats with CHF. The therapeutic effect of SXBXP may be related to better blood supply for myocardium and up-regulation of b(1)-AR and a(1B)-AR, and down-regulation of a(1A)-AR and b(3)-AR. The results show that SXBXP can be used in treatment of CHF and the therapeutic effect of large-dose SXBXP is superior to small-dose SXBXP.

Animals ; Blotting, Western ; Drugs, Chinese Herbal ; therapeutic use ; Echocardiography ; Heart Failure ; diagnostic imaging ; drug therapy ; Male ; Myocardial Infarction ; diagnostic imaging ; drug therapy ; Radiography ; Rats ; Rats, Wistar ; Receptors, Adrenergic, alpha-1 ; genetics ; metabolism ; Receptors, Adrenergic, beta ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction