BACKGROUND: Previous studies have found that polygonatum sibiricum polysaccharide (PSP) exhibits anti-osteoporosis effect, but its therapeutic effect in ovariectomized osteoporotic rats and the molecular mechanisms are poorly understood. OBJECTIVE: To investigate the effect of administration of PSP on the bone microstructure, bone mineral density as well as osteoblast- and osteoclast-related gene expression in rats. METHODS: Twenty-five infertile female Sprague-Dawley rats aged 3 months were randomly allotted into five groups (n=5 per group): sham operation (same volume normal saline), model, zoledronate (0.2 mg/kg?d), high-dose PSP (800 mg/kg?d) and medium-dose PSP (400 mg/kg?d) groups. All rats were subjected to ovariectomy except sham operation group. The administration was intragastrically given every 2 days beginning at 7 days after modeling and lasted 12 weeks. Then, the rats were sacrificed, and the uterus was weighed. The bilateral tibias were removed, one side for histomorphometric analysis by micro-CT, and the other one for RNA detection by qualified PCR. RESULTS AND CONCLUSION: Compared with the sham operation group, the rat body mass in the model group was significantly increased and the weight of uterus was significantly decreased (P < 0.05). Compared with the model group, zoledronate and high-dose PSP could significantly alleviate the excessive increase in body mass (P < 0.05). The bone mineral density in the model group was decreased by 63% compared with the sham operation group (P < 0.01), Compared with the model group, after 12-week high-dose PSP and zoledronate administration, the bone mineral density was increased by 44% and 38%, respectively (P < 0.01); the trabecular bone volume fraction and trabecular number rose significantly(P<0.05),while the trabecular separation decreased significantly(P<0.05).In vivo,PSP could significantly promote the expression levels of osteoblast-related genes (alkaline phosphatase, RUNX2, Col1a1 and osteocalcin), and significantly inhibit the expression levels of osteoblast-related genes (ACP5 and CTSK) (P < 0.05). These results imply that high-dose PSP can reduce bone loss and decrease of bone mineral density, improve the destruction of bone microstructure, as well as promote osteoblast-related genes but inhibit osteoclast-related gene mRNA expression in the ovariectomized rats.

Objective To explore the expression of protein arginine methyltransferases 6 (PRMT6) in chronic obstructive pulmonary disease (COPD) mouse model and its correlation with inflam mation gene interleukin 6 (IL-6) and cyclooxygenase 2 (COX-2).Methods Sixteen C57BL/8J mice were randomly divided into 2 groups:control group and cigarette smoke extract (CSE)-induced COPD group.Each group was injected intraperitoneally with phosphate-buffered saline solutions (PBS) or CSE at days 1,12,23 and measured lung function and collected lung tissue at day 29.The morphology change of the lung tissue was determined by hematoxylin and eosin (HE) stainning.The protein expression of PRMT6,H3R2me2a and H3K4me3 were detected in lung homogenates by Western-blotting.The mRNA expression of PRMT6,IL-6 and COX-2 were measured by quantitative real-time polymerase chain reaction (qRT-PCR).Results Comparing to control group,COPD group showed typical emphysema changes in the lung tissue,and significantly decreased lung function.The mRNA and protein expression of PRMT6 in the lung tissue of the mice with COPD were significantly decreased,following with the down-regulated signal level of H3R2me2a protein expression,while the increased level of IL-6 and COX-2 mRNA.Meanwhile,PRMT6 was negatively correlated with IL-6 and COX-2 mRNA expression.Conclusions PRMT6 was significantly reduced in CSE-induced COPD mouse model,following with decreased histone H3R2 dimethylation and increased H3K4 trimethylation,negatively correlating with inflammatory gene IL-6 and COX-2 transcription expression.PRMT6 downregulation may activate the transcriptional expression of inflammatory genes involved in the development of COPD,through the regulation of histone methylation level.

OBJECTIVETo evaluate the clinical efficacy of Jianpi Liqi Yiliu Formula (JLYF) combined with cytokine-induced killer (CIK) cells for treating patients with advanced hepatocellular carcinoma (HCC).

METHODSBetween January 2011 and January 2014, 60 advanced HCC patients were enrolled in this study, who were assigned to the treatment group and the control group according to their willingness for taking JLYF, 30 cases in each group. All patients received CIK cell treatment: 1 x 10⁹-3 x 10⁹ each time, by intravenous dripping from the 1st day to the 3rd day, once per day. Besides, patients in the treatment group took JLYF decoction, while those in the control group took Chinese medical decoction by syndrome typing. All patients received treatment of at least two cycles. The time to progression (TTP) , overall survival (OS), disease control rate (DCR), performance status scale (PS), Child-Pugh scale, and adverse reactions were observed, and subgroup analyzed.

RESULTSTo May 31, 2014, all patients reached the clinical endpoint. TTP was 3.5 months (95% Cl: 3.30-4.10) in the treatment group, better than that (2.5 months, 95% CI: 2.32-2.68) of the control group (P < 0.05). DCR was 36.7% in the treatment group and 30.0% in the control group (P > 0.05). OS was 5.2 months (95% CI: 4.53-5.87) in the treatment group and 4.6 months (95% CI: 4.06-5.14) in the control group (P > 0.05). The PS scale was 1.60 ± 0.10 after treatment, lower than that (1.80 ± 0.09) before treatment in the treatment group (P < 0.05). When the PS scale was 0-2 or Child-Pugh scale was class A, TTP was longer in the treatment group than in the control group (P < 0.05). No adverse reaction occurred in the two groups during the treatment course.

CONCLUSIONSThe combination of JLYF with ClK cell treatment could prolong advanced HCC patients' TTP, improve PS scale, as compared with syndrome typed Chinese medical decoction treatment group. Besides, when the PS scale was 0-2 or Child-Pugh scale was class A, it was a better treatment program for advanced HCC patients.

Carcinoma, Hepatocellular ; therapy ; Cell- and Tissue-Based Therapy ; Cytokine-Induced Killer Cells ; cytology ; Disease Progression ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Liver Neoplasms ; therapy

PURPOSE: The purpose of this study was to investigate the function of Zinc finger protein 488 (ZNF488) in nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: The endogenous expression of ZNF488 in NPC tissues, normal nasopharyngeal epithelium tissues and NPC cell lines were detected by quantitative reverse transcription polymerase chain reaction. ZNF488 over-expressing and knock-down NPC cell line models were established through retroviral vector pMSCV mediated over-expression and small interfering RNA (siRNA) mediated knock-down. The invasion and migration capacities were evaluated by wound healing and transwell invasion assays in ZNF488 over-expressing and control cell lines. Soft-agar colony formation and a xenograft experiment were performed to study tumorigenic ability in vitro and in vivo. Immunofluorescence and western blotting analysis were used to examine protein changes followed by ZNF488 over-expression. Microarray analysis was performed to explore gene expression profilings, while luciferase reporter assay to evaluate the transcriptive activity of Tcf/Lef. RESULTS: ZNF488 was over-expressed in NPC tissues compared with normal tissues, especially higher in 5-8F and S18, which are well-established high metastatic NPC clones. Functional studies indicate that over-expression of ZNF488 provokes invasion, whereas knock-down of ZNF488 alleviates invasive capability. Moreover, over-expression of ZNF488 promotes NPC tumor growth both in vitro and in vivo. Our data further show that over-expression of ZNF488 induces epithelial mesenchymal transition (EMT) by activating the WNT/beta-catenin signaling pathway. CONCLUSION: Our data strongly suggest that ZNF488 acts as an oncogene, promoting invasion and tumorigenesis by activating the Wnt/beta-catenin pathway to induce EMT in NPC.
Blotting, Western ; Carcinogenesis* ; Cell Line ; Clone Cells ; Epithelial-Mesenchymal Transition* ; Epithelium ; Fluorescent Antibody Technique ; Gene Expression Profiling ; Heterografts ; Luciferases ; Microarray Analysis ; Oncogenes ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Small Interfering ; Wnt Signaling Pathway* ; Wound Healing ; Zidovudine ; Zinc Fingers

Objective To explore the effects of dynamic axial compressive strain on the mRNA expression of bone formation related-genes in osteoblasts seeded in 3D silk fibroin scaffolds. Methods In the experimental group, MC3T3-E1 cells were seeded in 3D scaffolds and then subjected to dynamic axial compressive strain (at amplitude of 5% and frequency of 1 Hz, 30 min/day for 20 days), while in the control group, MC3T3-E1 cells were cultured statically without any mechanical stimulation. The gene expression of alkaline phosphatase (ALP), collagenⅠ(COL-Ⅰ), runt-related transcription factor 2 (Runx2), osterix (Osx), osteocalcin (OCN) was detected by real-time PCR. Results Under cyclic axial compressive strain, the Runx2, Osx and COLⅠmRNA levels increased by 280%, 68.9% and 79.6%, respectively, while the ALP and OC mRNA levels also up-regulated by 10.7% and 26.9%, respectively. There were significant differences in mRNA expression of osteogenesis-related genes between the experimental group and control group (P＜0.05). Conclusions Under the stimulation of cyclic axial compressive strain, the osteogenic differentiation of osteoblasts seeded in the silk fibroin scaffolds is promoted, with a significant up-regulation in the mRNA expression of COLⅠ, Runx2, Osx and OCN, which suggests that the stimulation of compressive stress at physiologic magnitude could be one of important mechanisms in promoting fracture healing. This research finding will be important for the clinic application of mechanical stimuli-mediated cell therapy for bone defection.