OBJECTIVE:To investigate the paracrine effects of bone marrow mesenchymal stem cells on osteoblast biological function. METHODS:Bone marrow mesenchymal stem cells were isolated using standard Ficol-Paquedensity gradient centrifugation. Mesenchymal stem cellconditioned medium was prepared to cultivate osteoblasts, MG63. Proliferation of MG63 cells was analyzed by cellcounting kit-8. Migration of MG63 cells was analyzed by cellscratch method. Alkaline phosphatase activity of MG63 cells was analyzed by microplate test kit. Real-time PCR was performed to evaluate osteoblast differentiation markers, alkaline phosphatase, col agen type I and osteocalcin. Alizarin red staining was performed to evaluate osteoblast mineralization. RESULTS AND CONCLUSION:The cells were strongly positive for CD44, CD73 and CD90, but negative for CD34. MG63 cells cultured in the conditioned medium showed better proliferation and migration than those cultured in the Dulbecco’s modified Eagle’s medium. The activity and mRNA expression of alkaline phosphatase were much higher after induction of 4, 7 days (P<0.01). There was no significant difference in expression of col agen type I and osteocalcin after induction of 4 days, but they were significantly higher than those in the control group after induction of 7 days (P<0.05). Alizarin red staining showed that the number of calcium nodules was increased and the mineral apposition was enhanced after induction of 21 days with the conditioned medium. These findings suggest that the paracrine substance of bone marrow mesenchymal stem cells can significantly promote osteoblast proliferation, migration, differentiation and mineralization.

Objective To investigate the effect of electroacupuncture on hypothalamic and hippocampal expressions of ghrelin and ghrelin receptor (GHS-R) mRNA in functional dyspepsia (FD) rats. Methods Eighty SD rats were randomly allocated to normal control, model control, medication and electroacupuncture groups, 20 rats each. A rat model of FD was made by tail clamp modeling method. The medication group received an oral gavage of cisapride solution and the electroacupuncture group, electroacupuncture treatment. The expressions of ghrelin protein and GHS-R mRNA in the hypothalamus and hippocampus were examined by Western blot and real-time PCR, respectively, in all the four groups. Results Compared with the model control group of rats, the expression of ghrelin protein in the hypothalamus and the expression of GHS-R mRNA in the hypothalamus and hippocampus increased in the medication group (P<0.05) and the expressions of ghrelin protein and GHS-R mRNA in the hypothalamus and hippocampus were up-regulated in the electroacupuncture group (P<0.05). Compared with the medication group of rats, the expressions of ghrelin protein and GHS-R mRNA in the hippocampus were up-regulated in the electroacupuncture group (P<0.05). Conclusions Electroacupuncture and medication can improve gastrointestinal functions and correct an imbalance in the brain-gut axis through the hippocampus-hypothalamic pathway by regulating hypothalamic and hippocampal ghrelin protein and GHS-R mRNA contents and activating hypothalamic neurons and hippocampal excitatory synapses.

OBJECTIVETo explore the effect of electroacupuncture (EA) on the expression of Ghrelin and mRNA expression of its receptor in functional dyspepsia (FD) rats.

METHODSTotally 80 rats were divided into the normal group, the model group, the drug therapy group, and the EA group according to random digit table, 20 in each group. FD model was duplicated by clipping tail modeling. Drug containing cisapride [2 mL/100 g, 0.09 g/(kg x d)] was administered to rats in the drug therapy group from the 3rd day after successful modeling, once per day. EA at Zusanli (ST36) (0.3-0.5 cun) and Taichong (LR3) (0.1-0.2 cun) was performed in the EA group. The twirling of needle was performed to the subsidence of needle, and then the needle was connected to HANS-200A Acupoint Nerve Stimulating Device using disperse-dense wave at 2 Hz, 2 mA, 30 min each time, once per day. Six days consisted of one therapeutic course, two courses in total with an interval of one day. The intestinal propulsive rate of ink was observed. Ghrelin protein expression in gastric tissue was detected by Western blot. mRNA expression of growth hormone secretagogue receptor (GHS-R) in stomach, hypothalamus, and hippocampus was detected using Real-time PCR respectively.

RESULTSCompared with the normal group, the intestinal propulsive rate of ink, Ghrelin protein expression in gastric tissue, mRNA expression of GHS-R in stomach, hypothalamus, and hippocampus decreased in the model group (P < 0.05, P < 0.01). Compared with the model group, the intestinal propulsive rate of ink, Ghrelin protein expression in gastric tissue, mRNA expression of GHS-R in stomach, hypothalamus, and hippocampus increased in the EA group (P < 0.01); mRNA expression of GHS-R in stomach, hypothalamus, and hippocampus increased in the drug therapy group (P < 0.01). Compared with the drug therapy group, Ghrelin protein expression in gastric tissue, mRNA expression of GHS-R in hypothalamus increased in the EA group (P < 0.05, P < 0.01).

CONCLUSIONEA could regulate Ghrelin content and GHS-R mRNA expression of FD rat hypothalamus, hippocampus, and gastric tissue, and promote the intestinal propulsive rate of ink.

Acupuncture Points ; Animals ; Dyspepsia ; metabolism ; therapy ; Electroacupuncture ; Ghrelin ; metabolism ; Hippocampus ; metabolism ; Hypothalamus ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Receptors, Ghrelin ; metabolism ; Stomach ; metabolism

Coding region instability determinant (CRD) is one of the influence factors of oncogene c-myc.Coding region instability determinant-binding protein (CRD-BP) can connect with CRD in order to protect CRD from nuclease attack,prevent rapid degradation of c-myc mRNA,and increase c-myc protein content.Beta-transducin repeats-containing protein (β-TrCP) can affect cell growth,differentiation,apoptosis and oncogenesis by regulating multiple signaling pathways and cell cycle.The overexpression of CRD-BP can upregulate the expression of β-TrCP and both of them play important roles in the tumorigenesis,progression,metastasis and invasion of colorectal cancer.

Objective To investigate effect of polymorphism of apolipoprotein E (APOE) gene on early apoptosis of astrocytes after hypoxic injury.Methods Astrocytes separated from APOE wild mice and APOE transgenic mice (ε3,ε4) were primarily cultured,and then purified and identified.Models of astrocyte hypoxic injury were set up by hypoxia.Morphological changes of astrocytes and mitochondria were observed by transmission electron microscope.Early apoptosis rate and changes of mitochondrial membrane potential were detected by flow cytometry.Results Cell foot process tumidness and mitochondria with irregular outline,vacuoles and irregular cristae were observed in each group by electron microscopy at six hours after hypoxia.There were no significant differences of cellular form changes among groups.Early apoptosis rate and decreasing degree of mitochondrial membrane potential in APOFε4 group were significantly higher than those in APOEε3 group and APOE wild group (P ＜ 0.05).Conclusion Compared with astrocytes from APOEε3 group and APOE wild group,mitochondrial membrane potential in astrocytes from APOEε4 group at early period after hypoxia declines more significantly,as may be one of causes for more astrocyte apoptosis.

Objective To reach parameter on sterilization wrap of medical instrument. Methods The parameter of one hundred twenty sterilization wraps with size 30 cm?40 cm?60 cm and weight 20kg was tested by Belimed sterilizer. Results There are 155 masculine gender and 445 negative in 600 quick reading biological indicators of 120 sterilization wraps and 53 masculine in bacteria detection, 9 cases failure in 1243 indicator, 52 wet wraps. Conclusion The parameter was to take three times negative pressure pulse and five times pressure pulse, sterilization temperature 134℃, sterilization time 11min and dry time 11min, matching to sterilization request of "Technical Standard For disinfection".

Adult ; Antibodies, Monoclonal, Murine-Derived ; administration & dosage ; therapeutic use ; Female ; Hemophilia A ; drug therapy ; Humans ; Rituximab

Objective To observe the regulatory effects of the preventive acupuncture and moxibustion at Guanyuan (RN4) points in ovariectomized rats, and discuss the mechanism of preventive acupuncture and moxibustion on hyoathalamus-pituitary-ovary (HPO) axis. Methods Totally 48 female SD rats were divided into 5 groups, including normal group, sham operation group, ovariectomized rats group, preventive acupuncture group and preventive moxibustion group. Except normal group and sham operation group, rats were ovariectomized after preventive acupuncture and preventive moxibustion for 30 d in corresponding groups. ELISA was used to determine the levels of E2, ER, GnRH, FSH and LH in the tissues including hypophysis, hypothalamus and uterus. Results Compared with the normal group, E2, ER and GnRH level in the tissue of the model group was significantly lower (P<0.01), the levels of FSH and LH increased obviously (P<0.01). Compared with the model group, E2, ER and GnRH level of preventive acupuncture group and preventive moxibustion group increased in different degree (P<0.05). FSH and LH level reduced in different degree (P <0.05). There was no obvious difference between preventive acupuncture and preventive moxibustion group (P>0.05). Conclusions The preventive acupuncture and preventive moxibustion at Guanyuan (RN4) has benign modulation to disordered HPO axis of ovariectomized rats. The ability of preventive acupuncture and preventive moxibustion are almost the same.

OBJECTIVE: To develop an HPLC method for the determination of Gastrodin in Qianglitianmaduzhong capsule. METHODS: The samples were separated on Diamond C18 column (250 mm?4.6 mm,5?m).The mobile phase consisted of acetonitrile - 0.1% phosphoric acid(2∶98) at a flow rate of 1.0 mL?min-1.The UV detection wave length was set at 210 nm and the column was kept at room temperature. RESULTS: The linear range of Gastrodin was 0.126 4～3.160 0 ?g(r=0.999 8), and the average recovery was 98.30%(RSD=0.8%,n=6).CONCLUSION: The method is suitable for the determination of gastrodin in Qianglitianmaduzhong capsule.

Objective:To evaluate the impact of altered collagen Ⅱ(CⅡ) peptide(S268-270) on T cell activation.Methods:Altered CⅡ263-272 with G268,P269 and K270 consecutively substitution with A or G were synthesized using solid phase various concentrations were added to HLADR1 transfected APC.Expression of FITC stainning was analyzed by fluorescence-activated cell sorting,and the binding of altered CⅡ peptide to HLA-DR1 molecule on cell membrane was evaluated.Irridiated HLA-DR1 expressing APC was incubated with CⅡ263-272 or hemagglutinin(HA)306-318 and altered CⅡ peptides at various concentrations for 2 hours before T cells(3.19) were added.Supernatant was harvested and then added to IL-2 dependent CTLL cell.The cell proliferation was examined by methotetrazolium(MTT) method.Results:The altered CⅡ peptide and wild type of CⅡ263-272 were able to competitively bind to HLA-DR1 molecule expressed on cell membrane of transfected APC.CⅡ or HA induced T cell activation was significantly inhibited by the altered peptide S268-270.Conclusion:Altered CⅡ263-272 with G268,P269 and K270 consecutively substitutions with A or G inhibited CⅡ263-272 and HA306-318 induced T cell activation through competitively binding to HLA-DR1,suggesting a new approach in inhibition of T cell activation in RA.