Genes, Reporter ; Genetic Vectors ; Luciferases ; genetics ; Transcription, Genetic ; bcl-2-Associated X Protein ; genetics

Aim To explore the effects of salvianolic acid B(SAB) and tanshinone ⅡA(TA) alone or the compatibility of these two effective components on the proliferation of rat vascular adventitial fibroblasts, and to observe the effects of the compatibility of the two on cell proliferation with the stimuation of angiotensin Ⅱ(Ang Ⅱ).Methods The effects of SAB and TA alone or the compatibility of the two on cell proliferation were detected by methyl thiazolyl tetrazolium(MTT) method, and flow cytometry was adopted to detect cell cycle with or without the induction of Ang Ⅱ.Results It was shown that SAB and TA alone could inhibit fibroblast proliferation in different degree.Through a series of concentration screening, three effective concentrations were obtained respectively and then the inhibition of cell growth was detected by Pairwise compatibilities.The results showed that the compatibility of TA(10-8 mol·L-1) and SAB(10-5 mol·L-1) had the most significant inhibitory effect(P<0.01), and they could inhibit cell proliferation, further flow cytometry was adopted to detect drug effects on cell cycle, the results indicated that the compatibility of SAB and TA mainly blocked the cells in G0/G1 phase.Induced by Ang Ⅱ, the compatibility of SAB and TA group, compared with Ang Ⅱ group, blocked thee cells in G0/G1 phase;and compared with combination of SAB and TA group, it mainly blocked cell cycle in S phase.Conclusion SAB and TA have certain inhibitory effect on fibroblast proliferation, also they could inhibit the proliferation induced by Ang Ⅱ, mainly by blocking the cells in G0/G1 phase.

BACKGROUND: Incidence rate of cervical osteoarthritis in the middleaged and elderly is high. Some researches on risk factor causing cervical osteoarthritis have been performed abroad, but most of the factors are being discussed.OBJECTIVE: To investigate the occurring cervical osteoarthritis risk factors in the middle-aged and elderly from different regions of China and provide evidences for prevention and intervention of cervical osteoarthritis in community.DESIGN: Cross-sectional study.SETTING: Beijing Hospital, Ministry of Health, together with Second Hospital of Xi'an Jiantong University, Institute of Integrated Traditional and Western Medicine of Hebei Medical University, Shanghai Huadong Hospital, Nanfang Hospital, Second Affiliated Hospital of Harbin Medical University, West China Hospital of Sichuan University.PARTICIPANTS: The investigation was conducted from July to August2005. On the basis of stratified multi-stage cluster sampling method, 6 218formal registered permanent residents of over 40 years old from Xi'an,Shijiazhuang, Shanghai, Guangzhou, Harbin and Chengdu were enrolled.They all agreed to join the investigation voluntarily. There were 2 916males of 40-94 years and 3 302 females of 40-86 years.METHODS: Questionnaire investigation of epidemiology of cervical osteoarthritis was performed in the testees, and radiograph was used in the persons with clinical symptom. The basic sample unit was neighborhood committee (city) and village committee (countryside). Sampling method:Taking each city as a whole, composed of two levels, namely city and countryside, in the first phase the persons were extracted from district (county),in the second phase from sub-district (countryside), in the third phase from neighborhood committee (village eommittee). Diagnosis standard of cervical osteoarthritis was positive clinical symptom and 2 grade or above of radiograph Kellgren & Lawrence grading. The content of questionnaire contained 6 aspects: general condition, history of present illness, history of past illness, physical check-up, radiographs and disease diagnosis, totally94 questions and 141 variation indexes. Influential factors of prevalence rate of cervical osteoarthritis were analyzed using multifactor non-conditional Logistic regression analysis. Odds ratio (OR) was used for expressing index of strength of relationship between disease and exposures. If OR ＞ 1,it was indicated that there was positive correlation between disease occurrence and exposures. If OR ＜ 1, it was suggested that there was negative correlation between disease occurrence and exposures.MAIN OUTCOME MEASURES: Prevalence rate of cervical osteoarthritis in each city and OR.RESULTS: Totally 6 218 investigational subjects were included in the result analysis, without drop out. ①Total prevalence rate of cervical osteoarthritis in population of 40 years or above from 6 domestic cities was23.6%. There was significnat difference of prevalence rate in each city (P＜0.01). ②Result of Logistic regression analysis: Age (OR=1.010-1.058),defecation with squat ting pot (OR =1.024-1.997) and history of hypertension (OR =1.815-3.078) were common risk factor in most areas. In northern area the common risk factor compos ed of daily stair climbing or grade climbing (OR =1.018-1.020), while drinking colored wine (OR=3.451, Xi'an), history of osteoarthri tis of father (OR =2.491, Xi'an), history of diabetes (OR =5.013, Shijiazhuang), history of osteoarthritis of mother (OR =2.045, Shanghai), smoking (OR =6.857, Guangzhou), age of starting drinking (OR =3.044, Guangzhou) and full-time athletic sports (OR=9.020, Harbin), etc. emerged in different areas.CONCLUSION: The onset of cervical osteoarthritis has the same risk factor in 6 domestic areas, and main risk factor in different areas has certain differences, which can provide reference data for the prevention and cure of cervical osteoarthritis for the future and reduce waster of medical resources.

OBJECTIVETo investigate the anti-tumor immune response of dendritic cells (DCs) acquiring antigens from apoptotic cholangiocarcinoma cells and their therapeutic effects on cholangiocarcinoma cells.

METHODSDCs from human peripheral blood monocytes which acquired antigen capturing and processing capacity, characteristics of maturation, were established in vitro using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then cholangiocarcinoma cells were induced to apoptosis with mitomycin. The three groups included (1) coculture of DCs, apoptotic cancer cells and T cells, (2) coculture of DCs, necrotic cancer cells and T cells, (3) coculture of DCs, cultured cancer cells and T cells. After 7 days, DCs and T cells were riched separately to perform anti-tumor cells test and immune response test.

RESULTSthese cells had typical dendritic cell morphology, expressed high levels of CD1a and B7, acquired antigen from apoptotic cells caused by mitomycin and could stimulate T cells to inhibit, even kill cholangiocarcinoma cells.

CONCLUSIONSThe DCs from peripheral blood monocytes induced by GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by mitomycin, and stimulate T cells activity obviously. It maybe become an effective therapy for tumor.

Antigens, Neoplasm ; immunology ; Apoptosis ; drug effects ; Bile Duct Neoplasms ; immunology ; pathology ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; immunology ; pathology ; Coculture Techniques ; Dendritic Cells ; drug effects ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; Mitomycin ; pharmacology ; T-Lymphocytes ; immunology ; Tumor Cells, Cultured

OBJECTIVETo investigate the sustaining effects of gene-transferring hepatic stellate cell strain CFSC/HGF on the development of hepatocytes.

METHODSA CFSC/HGF strain, expressing HGF steadily and effectively was established by recombined retroviral vector pMSCV-HGF infection. Morphology and ultra structure of hepatocytes, albumin and urea production, as well as ICG uptake and excretion were studied continuously following the hepatocytes cultured on the CFSC/HGF feeder layers. Parallel group of collagen-dependent hepatocytes culturing and hepatocytes culturing on CFSC were also conducted. Semi-quantitative RT-PCR analysis was made to evaluate the expression of HGF receptor c-Met.

RESULTSThe hepatocytes cocultured on CFSC/HGF feeder layers had a higher survival rate, and the functions of albumin secretion and urea syntheses and ICG uptake and excretion, were superior to the other two culture methods. The result of RT-PCR indicated that the c-Met expressed on the CFSC/HGF coculturing hepatocytes was up-regulated 2.23 times.

CONCLUSIONGene-transferring hepatic stellate cell strain CFSC/HGF exhibited a remarkable sustaining effect on the hepatocytes development. The up-regulation of c-Met expressed on the surfaces of the hepatocytes induced by CFSC/HGF might play some part in this function.

Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Colony-Forming Units Assay ; Gene Transfer Techniques ; Hepatocyte Growth Factor ; metabolism ; Hepatocytes ; cytology ; Humans ; Liver ; cytology ; Proto-Oncogene Proteins c-met ; metabolism ; Transfection

OBJECTIVETo study the effect of Aidi Injection (艾迪注射液,ADI) applied in the bronchial artery, applied in the bronchial artery infused (BAI) neo-adjuvant chemotherapy for stage III A non-small cell lung cancer (NSCLC) before surgical operation.

METHODSThe 60 patients with NSCLC stage III A underwent two courses BAI chemotherapy before tumor incision were assigned to two groups, the treatment and the control groups, using a random number table, 30 in each group. ADI (100 mL) was given to the patients in the treatment group by adding into 500 mL of 5% glucose injection for intravenous dripping once daily, starting from 3 days before each course of chemotherapy, and it lasted for 14 successive days, so a total of 28 days of administration was completed. The therapeutic effectiveness and the adverse reaction that occurred were observed, and the levels of T-lymphocyte subsets, natural killer cell activity, and interleukin-2 in peripheral blood were measured before and after the treatment.

RESULTSThe effective rate in the treatment group was higher than that in the control group (70.0% vs. 56.7%, P<0.05). Moreover, as compared with the control group, the adverse reaction that occurred in the treatment group was less and mild, especially in terms of bone marrow suppression and liver function damage (P<0.05). Cellular immune function was suppressed in NSCLC patients, but after treatment, it ameliorated significantly in the treatment group, showing significant difference as compared with that in the control group (P<0.05).

CONCLUSIONADI was an ideal auxiliary drug for the patients in stage III A NSCLC received BAI neo-chemotherapy before surgical operation; it could enhance the effectiveness of chemotherapy, ameliorate the adverse reaction and elevate patients' cellular immune function; therefore, it is worthy for spreading in clinical practice.

Adult ; Aged ; Antineoplastic Agents ; adverse effects ; pharmacology ; therapeutic use ; Bronchial Arteries ; drug effects ; pathology ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; immunology ; surgery ; Chemotherapy, Adjuvant ; Drugs, Chinese Herbal ; adverse effects ; pharmacology ; therapeutic use ; Female ; Humans ; Infusions, Intra-Arterial ; Injections ; Interleukin-2 ; blood ; Killer Cells, Natural ; drug effects ; immunology ; Lung Neoplasms ; blood ; drug therapy ; immunology ; surgery ; Lymphocyte Subsets ; drug effects ; immunology ; Male ; Middle Aged ; Neoplasm Staging ; Time Factors ; Treatment Outcome

OBJECTIVETo induce hepatic differentiation of human adipose-derived stem cells (hADSCs) in vitro.

METHODShADSCs were isolated from human adipose tissue and treated with improved hepatic medium containing HGF, bFGF and FGF4. After 7 days of culture, OSM was added to the culture media. Cell growth during hepatic differentiation was evaluated by CCK8 assay. Morphology of differentiation was examined under light microscope. Liver specific genes and proteins were detected by RT-PCR analysis and immunohistochemical staining, respectively. And functional characteristics of hepatocytes were also examined.

RESULTSThe number of hADSCs cultured in the improved hepatic media was increased significantly in comparison to hADSCs cultured in control media from 5 days to 21 days (t=6.59, 8.69, 15.94 and 24.64, respectively, P<0.05). The hADSCs-derived hepatocyte-like cells exhibited hepatocyte morphology, expressed hepatocyte markers, possessed hepatocyte-specific activities, such as uptake and excretion of indocyanine green, glycogen storage and albumin production.

CONCLUSIONhADSCs can be induced into hepatocyte-like cells in this differentiation system. And this differentiation system promoted the growth of hADSCs.

Adipose Tissue ; cytology ; Albumins ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cell Separation ; Cells, Cultured ; Culture Media ; Fibroblast Growth Factor 2 ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; Reverse Transcriptase Polymerase Chain Reaction ; alpha-Fetoproteins ; metabolism

OBJECTIVETo explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells (BMSCs) in vivo.

METHODSMelanocytes were isolated from human foreskin. BMSCs were isolated from human bone marrow. Both of them were co-cultured at a ratio of 1:10, and then were implanted into the collagen membrane to construct the tissue-engineered skin, which was applied for wound repair in nude mice. The effectiveness of wound repair and the distribution of melanocytes were evaluated by morphological observation, in vivo 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.

RESULTSThe wounds were satisfactorily repaired among the nude mice. The melanocytes were distributed in the skin with normal structure, as confirmed by DAPI fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy.

CONCLUSIONMelanocytes and BMSCs, after proper in vitro culture at an appropriate ratio, can construct the tissue-engineered skin with I type collagen membrane.

Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Coculture Techniques ; Collagen Type I ; Humans ; Melanocytes ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Nude ; Skin ; injuries ; Skin, Artificial ; Tissue Engineering

OBJECTIVETo investigate the effects of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD(34)(+) hematopoietic stem/progenitor cells in cord blood.

METHODSRetroviral vectors containing Tpo or IL-11 gene were constructed and used to transfect the stromal cell line HFCL. Tpo and/or IL-11 mRNA was assayed by Northern blot. Non-modified stromal cells were used, CD(34)(+) hematopoietic stem/progenitor cells from cord blood were expanded on gene-modified stromal cells for 7 days. The phenotype of CD(34)(+)CD(38)(-) primitive progenitors was detected by flow cytometry.

RESULTSHFCL expressed Tpo and/or IL-11 mRNA after transfected by the retroviral vectors. The percentages of CD(34)(+)CD(38)(-) primitive progenitors in the cultures of Tpo, IL-11 and Tpo + IL-11 modified HFCL were (1.8 +/- 0.24)%, (1.62 +/- 0.23)%, and (2.45 +/- 0.28)%, respectively, which were higher than that in the control [(0.8 +/- 0.23)%].

CONCLUSIONThe stromal cells modified by Tpo and/or IL-11 gene were able to enhance ex vivo expansion of CD(34)(+) and CD(34)(+)CD(38)(-) hematopoietic stem/progenitor cells from cord blood.

ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Humans ; Infant, Newborn ; Interleukin-11 ; genetics ; Membrane Glycoproteins ; Stromal Cells ; physiology ; Thrombopoietin ; genetics

OBJECTIVETo identify genes that differentially expressed in Lin(-)CD(34)(-) and Lin(-)CD(34)(+) cells.

METHODSWith Lin(-)CD(34)(-) cells as tester and Lin(-)CD(34)(+) cells as driver, cDNA subtractive library for Lin(-)CD(34)(-) cells was constructed using suppression subtractive hybridization technique. Part of clones in the library were sequenced and the homologue analysis was conducted against the DNA database in GenBank.

RESULTS593 clones containing an average of 300 - 500 bp insert were identified. Of them, 53 randomly selected ESTs were sequenced. Homologue analysis revealed that 37 ESTs represented 10 known genes, and the other 16 ESTs represented 4 novel sequences.

CONCLUSIONPart of specifically expressed genes in Lin(-)CD(34)(-) cells were identified, which maybe related to Lin(-)CD(34)(-) cells' specific characteristics.

Antigens, CD34 ; metabolism ; Cloning, Molecular ; Gene Expression Profiling ; Gene Library ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; In Vitro Techniques ; Nucleic Acid Hybridization