[Intestinal dysfunction of spinal cord injury treated with tongdu tiaoshen moxibustion: a randomized controlled trial].

OBJECTIVE: To compare the clinical effect on intestinal dysfunction of spinal cord injury (SCI) between the comprehensive therapy of tongdu tiaoshen moxibustion (moxibustion for opening the governor vessel and regulating the spirit) and rehabilitation training and the simple treatment with rehabilitation training. METHODS: A total of 60 patients with intestinal dysfunction of SCI were randomized into a comprehensive therapy group and a rehabilitation group, 30 cases in each one (3 cases were dropped out in each group). On the base of the routine western medicine treatment and rehabilitation training, the bowel training and rectal function training were provided, once a day in the rehabilitation group. In the comprehensive therapy group, on the base of the treatment as the rehabilitation group, the tongdu tiaoshen moxibustion was exerted at Yaoyangguan (GV 3), Mingmen (GV 4), Zhiyang (GV 9), Dazhui (GV 14) and Baihui (GV 20), etc, once a day, 30 min each time. In both groups, the treatment for 4 weeks was as one course and 3 courses of treatment were required. Separately, before treatment, after 4, 8 and 12 weeks of treatment, the scores of neurogenic bowel dysfunction (NBD) and World Health Organization quality of life scale (WHOQOL-BREF) were observed and the clinical effect was evaluated after 12 weeks of treatment. RESULTS: After treatment, the total effective rate was 88.9% (24/27) in the comprehensive therapy group, which was higher than 74.1% (20/27) in the rehabilitation group (P<0.05). After 4, 8 and 12 weeks of treatment, NBD scores were all reduced obviously as compared with those before treatment in the two groups (all P<0.01). After 8 and 12 weeks of treatment, NBD scores in the comprehensive therapy group were lower than the rehabilitation group (both P<0.05). After 4, 8 and 12 weeks of treatment, the scores of all of the domains (psychology, physiology, social relations and environment) in WHOQOL-BREF were higher than those before treatment in the two groups (all P<0.01). After 4 weeks of treatment, the scores in the psychology and physiology domains in the comprehensive therapy group were higher than the rehabilitation group (all P<0.05). After 8 and 12 weeks of treatment, the scores of all of the domains in the comprehensive therapy group were higher than the rehabilitation group (all P<0.05). CONCLUSION: The comprehensive treatment of tongdu tiaoshen moxibustion and rehabilitation training achieves the better effect on intestinal dysfunction of SCI than the simple rehabilitation training and greatly improves the quality of life in SCI patients.

[Potential Value of Cancer-Associated Fibroblasts in Hematological Malignancies--Review].

Cancer-associated fibroblasts (CAF), as one of the most important components of tumor microenvironment, which plays important role in tumorigenesis, development, infiltration and metastasis of cancers. In a variety of solid tumors, CAF can even determine the fate of tumor cells. In view of its pivotal role in promoting tumor progression, CAF has recently become a therapeutic target for a variety of tumors. However, there are a few studies on CAF in hematological malignancies. Recent studies have found that the resistance, relapse of AML, MM, CLL and myelofibrosis of MPN closely relate with CAF, so targeting CAF can effectively enhance the killing effect of chemotherapy drugs on tumor cells, thus improve the efficacy, CAF is expected to become a new target for the treatment of hematological malignancies. This review summarizes recent advances in cancer-associated fibroblasts in hematological malignancies.

Knockdown of AGR2 induces cell apoptosis and reduces chemotherapy resistance of pancreatic cancer cells with the involvement of ERK/AKT axis.

BACKGROUND: Pancreatic cancer (PC), an aggressive human malignancy, presents with a striking resistance to chemotherapy. Interesting, AGR2 has been found to be upregulated in various cancers and has been found to promote the dissemination of PC cells. Thereby, a series of in-vitro experiments were performed to investigate the relationship between AGR2 and the ERK/AKT axis, and to explore whether it affects PC cells. METHODS: Positive expression of AGR2 protein in the PC and paracancerous tissues collected from 138 patients with PC was detected using immunohistochemistry. After treatment with FGF2 (an ERK/AKT axis agonist), siRNA against AGR2 or their combination respectively, cell viability, chemotherapy resistance, radiotherapy resistance, migration, invasion and apoptosis in PC cells were detected using CCK8 assay, MTT assay, clone formation assay, wound healing assay, Transwell assay and flow cytometry, respectively. The expressions of AGR2 and ERK/AKT axis-related genes and proteins in tissues and cells were detected using reverse transcription quantitative polymerase chain reaction and Western blot assay. RESULTS: PC tissues exhibited highly-expressed AGR2 and abnormally activated ERK/AKT axis. FGF2 promoted the expression of AGR2, ERK/AKT axis activation, cell viability, chemotherapy resistance, migration and invasion, but decreased cell apoptosis in PC cells. However, knockdown of AGR2 resulted in inhibition of the ERK/AKT axis, reduced PC cell viability, chemotherapy resistance, migration and invasion but increased cell apoptosis in PC cells. CONCLUSION: The findings reveal that AGR2 silencing could promote cell apoptosis and inhibit cell migration, invasion and chemotherapy resistance of PC cell with the involvement of the ERK/AKT axis.

Lentiviral-mediated short hairpin RNA silencing of APE1 suppresses hepatocellular carcinoma proliferation and migration: A potential therapeutic target for hepatoma treatment.

Apurinic/apyrimidinic endonuclease-1 (APE1) is a protein involved in DNA repair and transcriptional regulation of gene expression. APE1 expression was reported to be correlated with poor prognosis in hepatocellular carcinoma (HCC) patients. Based on our previous study, we hypothesized that APE1 may be involved in the metastatic progression of HCC. Thus, the present study aimed to investigate the knockdown effect of APE1 using shRNA in HCC and demonstrate that silencing of APE1 in MHCC97-H cells can decrease the oncogenic transforming potential in vitro and reduce the growth of HCC tumor xenografts in vivo. Silencing of APE1 expression decreased the cell proliferation and survival, reduced the cell adhesion ability in Matrigel or fibronectin-coated plates and suppressed the cell migration and invasion in a Transwell assay of HCC cells. In the xenograft study, tumor growth was markedly inhibited in the APE1-silenced group. Silencing of APE1 in MHCC97-H cells decreased the oncogenic transforming potential in vitro and reduced the growth of HCC tumor xenografts in vivo. Inhibition of APE1 may present a novel therapeutic approach for the treatment of HCC.

Effect of serum choice on replicative senescence in mesenchymal stromal cells.

BACKGROUND AIMS: Multipotent mesenchymal stromal cells (MSCs) are promising candidates for innovative cell therapeutic applications. Before their use, however, they usually need to be expanded in vitro with serum-supplemented media. MSCs can undergo replicative senescence during in vitro expansion, but it is not yet clear how serum supplements influence this process. METHODS: In the present study, we compared how media supplemented with fetal bovine serum (FBS) or calf serum (CS) affected morphology, proliferation, differentiation, senescence and other functional characteristics of human umbilical cord-derived MSCs (UC-MSCs). RESULTS: UC-MSCs cultured in both FBS- and CS-containing media were able to differentiate along osteogenic and adipogenic lineages but ultimately reached proliferation arrest. However, senescence-associated characteristics, such as ß-galactosidase activity, reactive oxygen species levels, proliferation rate and gene expression, demonstrate that UC-MSCs grown with FBS have better proliferation potential and differentiation capacity. In contrast, UC-MSCs grown with CS have a higher proportion of apoptotic cells and senescent characteristics. Possible mechanisms for the observed phenotypes include changes in gene expression (Bax, p16, p21 and p53) and cytokine production (interleukin-6 and interleukin-8). CONCLUSIONS: This study demonstrates that FBS-supplemented media provides a better microenvironment for the expansion of UC-MSCs in vitro than CS-supplemented media. This work provides insight into MSCs generation practices for use in basic research and clinical therapies.

Rutaecarpine inhibits angiotensin II-induced proliferation in rat vascular smooth muscle cells.

OBJECTIVE: To evaluate the effects and possible mechanisms of rutaecarpine on angiotensin II (Ang II)-induced proliferation in cultured rat vascular smooth muscle cells (VSMCs). METHODS: VSMCs were isolated from Male Sprague-Dawley rat aorta, and cultured by enzymic dispersion method. Experiments were performed with cells from passages 3-8. The cultured VSMCs were randomly divided into control, model (Ang II 0.1 µmol/L), and rutaecarpine (0.3-3.0 µmol/L) groups. VMSC proliferation was induced by Ang II, and was evaluated by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay and cell counting. To examine the mechanisms involved in anti-proliferative effects of rutaecarpine, nitric oxide (NO) levels and NO synthetase (NOS) activity were determined. Expressions of VSMC proliferation-related genes including endothelial nitric oxide synthase (eNOS), and c-myc hypertension related gene-1 (HRG-1) were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Rutaecarpine (0.3-3.0 µmol/L) inhibited Ang II-induced VSMC proliferation and the best effects were achieved at 3.0 µmol/L. The Ang II-induced decreases in cellular NO contents and NOS activities were antagonized by rutaecarpine (P <0.05). Ang II administration suppressed the expressions of eNOS and HRG-1, while increased c-myc expression (P <0.05). All these effects were attenuated by 3.0 µmol/L rutaecarpine (P <0.05). CONCLUSION: Rutaecarpine is effective against Ang II-induced rat VSMC proliferation, and this effect is due, at least in part, to NO production and the modulation of VMSC proliferation-related gene expressions.

Evaluation of vaporized hydrogen peroxide fumigation as a method for the bio-decontamination of the high efficiency particulate air filter unit.

OBJECTIVE: To evaluate the performance of vaporized hydrogen peroxide (VHP) for the bio-decontamination of the high efficiency particulate air (HEPA) filter unit. METHODS: Self-made or commercially available bioindicators were placed at designated locations in the HEPA filter unit under VHP fumigation. The spores on coupons were then extracted by 0.5 h submergence in eluent followed by 200- time violent knocks. RESULTS: Due to the presence of HEPA filter in the box, spore recovery from coupons placed at the bottom of the filter downstream was significantly higher than that from coupons placed at the other locations. The gap of decontamination efficiency between the top and the bottom of the filter downstream became narrower with the exposure time extended. The decontamination efficiency of the bottom of the filter downstream only improved gently with the injection rate of H2O2 increased and the decontamination efficiency decreased instead when the injection rate exceeded 2.5 g/min. The commercially available bioindicators were competent to indicate the disinfection efficiency of VHP for the HEPA filter unit. CONCLUSION: This assay developed can detect all 16 ß-lactams demanded by the European Union (EU). The whole procedure takes only 45 min and can detect 42 samples and the standards with duplicate analysis.

Decontamination of Bacillus subtilis var. niger spores on selected surfaces by chlorine dioxide gas.

OBJECTIVE: Chlorine dioxide (CD) gas has been used as a fumigant in the disinfection of biosafety laboratories. In this study, some experiments were conducted to assess the inactivation of spores inoculated on six materials [stainless steel (SS), painted steel (PS), polyvinyl chlorid (PVC), polyurethane (PU), glass (GS), and cotton cloth (CC)] by CD gas. The main aims of the study were to determine the sporicidal efficacy of CD gas and the effect of prehumidification before decontamination on sporicidal efficacy. METHODS: Material coupons (1.2 cm diameter of SS, PS, and PU; 1.0 cm×1.0 cm for PVC, GS, and CC) were contaminated with 10 µl of Bacillus subtilis var. niger (ATCC 9372) spore suspension in mixed organic burden and then dried in a biosafety cabinet for 12 h. The spores were recovered by soaking the coupons in 5 ml of extraction liquid for 1 h and then vortexing the liquid for 1 min. RESULTS: The log reductions in spore numbers on inoculated test materials exposed to CD gas [0.080% (volume ratio, v/v) for 3 h] were in the range of from 1.80 to 6.64. Statistically significant differences were found in decontamination efficacies on test material coupons of SS, PS, PU, and CC between with and without a 1-h prehumidification treatment. With the extraction method, there were no statistically significant differences in the recovery ratios between the porous and non-porous materials. CONCLUSIONS: The results reported from this study could provide information for developing decontamination technology based on CD gas for targeting surface microbial contamination.

[The effects of mesenchymal stem cells on the aging kidneys in rats].

OBJECTIVE: To study the effect of mesenchymal stem cells on the aging rat kidney and to explore the underlying mechanism. METHODS: Rat models of senile kidney were built with hypodermic injection of D-galactose daily. Injections of MSCs of 3 x 10(6) were given to each rat through vena caudalis and CFSE was used as a tracing label to detect the distribution of MSCs in vivo. After 24 h, rats were dissected and their kidneys were frozen for section. MSCs were observed with Fluophot and quantitative analysis of the various parameters of kidney was performed under a light microscope with BI2000 image analysis system. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and kidney were measured wtih hydroxylamine and chromatometry. The expression of VEGF and P16 mRNA in kidney tissue was detected with real-time PCR and Western blotting. RESULTS: MSCs was found homing in the rat kidney, and the glomerular size, sclerosis-rate and the average cell count of glomerulus in the treated group were different from those of the model group (P < 0.05). In the treated group, the activity of SOD was significantly higher and the content of MDA was lower in serum and kidney than that in the model control group (P < 0.05). The expression of VEGF mRNA and protein in the kidneys of MSCs group increased significantly as compared with the model group (P < 0.05). The expression of P16 mRNA and protein in the kidney of MSCs group decreased significantly compared with the model group (P < 0.05). CONCLUSION: MSCs can increase the expression of VEGF while decrease the expression of P16, so as to play a key role in the anti-aging on rat kidney.

[In vitro tracing of rat bone marrow mesenchymal stem cells using the fluorescent dye CFSE].

OBJECTIVE: To determine the optimal condition for labeling rat mesenchymal stem cells (MSCs) using the fluorescent dye CFSE and the maximum time length allowed by CFSE staining for MSC tracing in vitro. METHODS: Rat MSCs were labeled with CFSE at different concentrations (2.5, 5.0, 10.0, 20.0 and 40.0 micromol/L) for 1, 5 or 10 min. The transfection efficiency and fluorescence intensity in the cells were measured by flow cytometry and fluorescence microscope to determine the optimal condition for MSC labeling. Under the optimal condition, the effect of CFSE on the growth of MSCs was evaluated by MTT assay, and flow cytometry and fluorescence microscope were used to determine the maximum time length following CFSE labeling to allow MSC tracing. RESULT AND CONCLUSION: Staining with CFSE at 20.0 micromol/L for 5 min was optimal for labeling rat MSCs in vitro, which allowed detection of the MSCs as long as 21 days after the labeling without obviously affecting the cell growth (P>0.05).

[Different cytotoxicity of NK cells against high and low expression of ATP-binding cassette transporter (ABCG(2)) of human multi-drug resistant nasopharyngeal carcinoma cells].

AIM: To investigate the different cytotoxicity sensitivity of high and low expression of ATP-binding cassette transporter (ABCG(2)) of human multi-drug resistant nasopharyngeal carcinoma CNE2/DDP cell line (ABCG(2)(High) cell and ABCG(2)(Low cell)) to NK cells. METHODS: ABCG(2)(High)CNE2/DDP cells, ABCG(2)(Low) CNE2/DDP cells and NK cells were isolated by magnetic activated cell sorting (MACS). The cytotoxic effects of NK cells against K562, ABCG(2)(High) and ABCG(2)(Low)CNE2/DDP cells were detected with LDH releasing assay. The apoptosis rate and expression of NKG2D-ligands on target cells were evaluated by FCM. RESULTS: The expression of ABCG(2) in ABCG(2)(High) CNE2/DDP and ABCG(2)(Low) CNE2/DDP cells was (91.40+/-2.32)% and (1.70+/-0.24)%, respectively. More than 90% of the isolated NK cells were CD3(-)CD16(+)CD56(+) cells. As for the cytotoxicity of NK cells against ABCG(2)(Low), ABCG(2)(High)CNE2/DDP and K562 cells at the E vs T ratio of 10 vs 1 and 20 vs 1, ABCG(2)(High) CNE2/DDP cells possessed highest cytotoxic sensitivity. The apoptosis rate of ABCG(2)(High) CNE2/DDP cells and ABCG(2)(Low)CNE2/DDP cells was (12.90+/-1.51)% and (6.13+/-0.85)%, respectively. The expression of five kinds of NKG2D ligand of ABCG(2);(High)CNE2/DDP cells is higher than that of ABCG(2)(Low)CNE2/DDP cells. CONCLUSION: ABCG(2)(High) CNE2/DDP cells are more sensitive to NK cells than ABCG(2)(Low) CNE2/DDP cells in cytotoxicity because the expression of NKG2D ligands of ABCG(2)(High)CNE2/DDP cells is higher than that of ABCG(2)(Low)CNE2/DDP cells, which results in the cytotoxic sensitivity of increased tumor cells to NK cells.

[Create mathematical model and analysis of correlation between Chinese medicinal characteristics and immunoregulatory activity based on literature informatics].

OBJECTIVE: Intends to create mathematical model and analysis of correlation between Chinese medicinal characteristics and immunoregulatory activity based on literature informatics. METHOD: The numbers of the Chinese medicines with immune effects were worked out within the framework of "The China Pharmacopeia" of 2005 edition, from the literature publicized since 1980. The correlation and mathematical model were figured out between Chinese medicinal characteristics including biological classification, different tastes, channel tropism as well as the parts used and immunoregulatory activity based on the statistical software SPSS. RESULT: The results showed that the immunoregulatory activity was related to the five tastes of Chinese medicines, and the pungent medicines had less immune effect. The Chinese medicines of underground parts had more immune effect compared with other parts of the medicine. Medicines acting upon heart and kidneys were more powerful as for the immune effects (P <0.05). The coincidence was 74.7% between mathematical computing and original classification. CONCLUSION: There are correlations,between Chinese medicinal characteristics and immunoregulatory activity. The mathematical model based on these results can be used for immunopharmacology.

[Application of fingerprint of active part on extraction techniques for Vernonia anthelmintica].

OBJECTIVE: To probe into feasibility of extraction techniques for Vernonia anthelmintica using HPLC fingerprint of active part as model. METHODS: The extraction techniques were studied by multi-index test experiment formula evaluation and orthogonal design, which took change of peak area in HPLC fingerprint of active part for Vernonia anthelmintica as indices. RESULTS: The optimum extracting procedure was as follows: using 10 times volume of water for each time, soaking one hour and then boiling for three times sustaining one hour each. CONCLUSION: It is scientific, simple and applicable.