BACKGROUND:Osteoprotegerin and nitrogen monoxidum play a key role in the prevention and treatment of osteoporosis. However, the correlation of the two in inhibiting the proliferation and differentiation of osteoclasts remains unclear.
OBJECTIVE:To observe the effect of different doses of osteoprotegerin on nitric oxide production and endothelial nitric oxide synthase activity in osteoclasts.
METHODS:Tartrate resistant acid phophatase staining was used to test whether the induced cells are osteoclasts. Osteoclasts were divided into six groups:blank control group (no reagent);negative control group (DMEM);four osteoprotegerin groups (10, 25, 50, 75μg/L of osteoprotegerin). Annexinv-FITC kit and flow cytometry were used to test the apoptosis rate of osteoclasts. Real-time PCR was used to detect the expression of osteoclasts marker gene, TRAP mRNA and protein kinase K mRNA. Nitric oxide production and nitric oxide synthase activity were determined using the corresponding kits. Four osteoprotegerin groups were added with L-NAME, a kind of nitric oxide synthase inhibitor, to test the changes of the apoptosis rates of osteoclasts and the changes of the expression of TRAP mRNA and protein kinase K mRNA of osteoclasts.
RESULTS AND CONCLUSION:Osteoprotegerin inhibited the differentiation of osteoclasts and induced the apoptosis. Osteoprotegerin concentration had a positive correlation with the apoptosis rate of osteoclasts, and a negative correlation with the numbers of osteoclasts and expression of marker gene TRAP mRNA and protein kinase K mRNA in osteoclasts. Osteoprotegerin boosted the nitric oxide production and endothelial nitric oxide synthase activity, and osteoprotegerin concentration was positively correlated to the nitric oxide production and endothelial nitric oxide synthase activity. After Raw264.7 cells were in vitro cultured, osteoprotegerin and nitric oxide play a synergic role in inhibiting osteoclasts production and promoting the apoptosis. We speculate that there is osteoprotegerin/endothelial nitric oxide synthase/nitric oxide signal pathway.

BACKGROUND:Stem cel transplantation has been shown to have many advantages in treatment of diseases. Its application can treat various diseases in many systems such as nervous system, immune system and endocrine system. Its advantage is incomparable. These are al credit to the special abilities, including easy sample colection, strong self proliferative ability, wide differentiation range, repair of injured tissue and immunoloregulation. OBJECTIVE:To explore the clinical effects of stem cel transplantation in treatment of nonunion. METHODS:A computer search was performed in CNKI database for articles concerning stem cel transplantation for nonunion published from 2005 to 2014. The key words were stem cels, nonunion in Chinese. 244 articles were analyzed, and typical literatures were further compared. RESULTS AND CONCLUSION:Stem cels have the abilities of self-renewal and differentiation. Their source is not limited, and the sample colection is convenient. Stem cels have smal injury to the donor, do not have immunogenicity, and have fewer complications. The cost is low. Stem cels are easily isolated and cultured, can proliferate infinitely, and provide a pathway for severe bone injury such as nonunion, bone defects and comminuted fracture. The deep study of stem cels creates a new direction for nonunion treatment, and has achieved certain results in clinical trials.

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 investigate the effect of Ginkgo biloba extract on serum divalent metal transporter1 ( DMT1 ) , glucose regulated protein 75(grp75) and nerve function in patients with Parkinson disease.Methods 38 cases of patients loith parkinson disease according to different drugs were divided into experimental group and control group, 19 cases in each group.Control group was treated with levodopa and Benserazide tablet, experimental group on the basis of control group, was given Ginkgo biloba extract tablets, treatment for 4 weeks.After treatment, DMT1, grp75 and cognitive function of all patients in substantia nigra were detected.ResuIts Compared with before treatment, two groups of patients with lower DMT1 level (P<0.05), compared with control group, experimental group of patients with lower DMT1 levels (P<0.05).Compared with pre-treatment, two groups of patients grp75 level was higher (P<0.05), compared with control group, experimental group after treatment grp75 level was higher (P<0.05).Compared with before treatment, the two groups of patients with MoCA scores were higher (P<0.05), HAMD scores were lower (P<0.05).Compared with control group, experimental group after treatment MoCA scores were higher (P<0.05), HAMD scores were lower (P<0.05).ConcIusion Ginkgo biloba extract can significantly reduce the level of DMT1 in the substantia nigra of Parkinson patients, increase the level of grp75, and improve the cognitive function.

Objective To establish a model fit for axonal regeneration research after its mechanical injury.Methods Cortical explants from mice were planted on culture dishes by microglass pipettes or routine glass pipettes.The cell body and dendrites in axonal area were detected by immunofluorescence and RT-PCR.Besides,purity of regenerated axons was also tested by immunofluorescence and RT-PCR after mechanical transection of axons under microscopy.Results Compared with explants planting by routine glass pipettes,in the outside 1/2 axons of explants planted by micro-glass pipettes,the immunofluorescence and RT-PCR showed negative nucleus and dendrites.In the regenerated axons following mechanical transection of explants planted by micro-glass pipette,the immunofluorescene and RT-PCR showed no regenerated axons nucleus mixed into the dendrites and nucleus.Conclusions Explants planted by micro-glass pipette obtains enough pure axons and regenerated axons.The establishment of models of axonal mechanical transection lays foundation for its molecular study after trauma.

ObjectiveTo establish a central nervous system axonal mechanical transection model in vitro and observe the axonal regeneration style and speed following the transection. MethodsThe cortical explants the mice were cultured in vitro,of which the axon and dendrite parts were marked using immunofluorescence.The model was built by mechanically transecting the axons and removing the severed axons.The axonal regeneration style was observed by tracing the undeveloped cellular bodies adhering to the residual axonal surface.The growth speed of the regenerated axons and normal axons were measured as well.Results ( 1 ) After mechauically transecting the axons of explants,the axonal regeneration was largely founded at the transected line.The undeveloped cellular bodies went over the transected line and spread to the distal area with axonai regeneration. (2) The axonal growth speed was (118 ± 32) μm/d and (72 ±41) μm/d in the transection group,but was (41 ± 17) μm/d and (32 ± 19) μm/d in the control group at 24 and 48 hours respectively. ConclusionThe regeneration style of the transected axons is the extension of the axonal stumps,rather than sprouting growth,and the growth speed is faster than that of the normal axons.

Objective To explore the feasibility of dual-source CT coronary angiography (CTCA) using 80 kV tube voltage setting in patients with low body mass index (BMI).Methods A total of 240 patients with suspected coronary artery disease (BMI range 18.6-21.5 kg/m2) underwent dual-source CTCA.They were randomly assigned to group A (120 kV tube voltage),B (100 kV tube voltage) and C (80 kV tube voltage) based on a random number table.The contrast media dose,volume CT dose index (CTDIvol),dose length product (DLP),and effective dose (ED) were evaluated for each patient.Image noise,CT value,contrast,signal-to-noise ratio (SNR),contrast-to-noise ratio (CNR) and figure of merit (FOM) of coronary artery were all evaluated.Tie image quality of coronary artery was assessed with a threepoint scale (3 points:excellent,2 points:general,1 point:poor).Scan length,CTDIvol,DLP,ED,and contrast media dose for patients,image noise,CT value,contrast,SNR,CNR,and FOM of coronary artery were all analyzed using one-way ANOV4 analysis for 3 groups.Time windows of ECG-pulsing protocol were analyzed using x2 test,and the image quality scores of coronary artery were analyzed using Kruskal-Wallis test.Results There were no differences in scan length and Time windows of ECG-pulsing protocol among 3 groups (scan length F =2.58,P ＞ 0.05,Time windows of ECG-pulsing protocol x2 =0.77,P ＞ 0.05).The average contrast media doses were (82.0 ± 6.4),(76.8 ± 6.1),and (59.1 ± 3.5) ml,the average CTDIvol were(27.5 ±6.2),(18.7 ±4.4),and(11.4 ±2.4)mGy,the average DLPs were (427.7 ±92.4),(295.4 ± 77.1),and (183.9 ± 41.3) mGy · cm,and the average EDs were (6.1 ± 1.3),(4.1 ± 1.1),and (2.6 ± 0.6) msy for group A,B and C,respectively.There were statistical differences in contrast media dose,CTDIvoI,DLP,and ED among 3 groups (the F values for contrast media dose,CTDIvol,DLP,and ED were 383.08,248.13,221.05,and 234.81,respectively,all P ＜0.01).Compared to group A and B,the contrast media dose in group C decreased 27.9％ and 23.0％,and the ED decreased 57.4％ and 36.6％,respectively.The average image noise was (22.6 ±2.2),(26.1 ±3.0),and(29.1 ±3.4)HU for group A,B and C,respectively,and there was statistical difference in image noise among 3 groups (F=101.32,P ＜0.01).The average CT value,contrast,SNR,CNR and FOM were (438.3 ± 66.3)HU,(517.3 ± 67.8)HU,19.5 ±2.8,23.0 ±3.0,and 92.9 ± 31.0 in proximal RCA and (440.2 ±59.9) HU,(509.5 ± 62.6) HU,19.6 ±2.6,22.7 ±2.9,and 90.1 ±29.7 in LMA for group A,(534.2 ± 68.8) HU,(628.9 ±70.0) HU,20.7 ± 3.2,24.3 ± 3.6,and 157.6 ± 59.8 in proximal RCA and (528.4 ± 61.2) HU,(607.9 ± 71.2) HU,20.4±3.0,23.5 ±3.4,and 147.6 ±57.6 in LMA for group B,and (602.1 ±78.8)HU,(696.8 ±83.3) HU,20.8 ± 2.9,24.1 ± 3.2,and 239.3 ± 74.8 in proximal RCA and (592.5 ± 72.3) HU,(671.8 ±82.5) HU,20.5 ± 2.5,23.2 ± 3.0,and 221.8 ± 65.7 in LMA for group C,respectively.The CT value,contrast,SNR,CNR and FOM of coronary artery for group C did not decrease (the F values for CT value,contrast,SNR,CNR and FOM were 106.06,119.90,4.69,3.70,and 127.50 in proximal RCA,and 111.79,101.57,2.68,1.39,and 123.00 in LMA,and the P values were ＜0.01,＜0.01,＜0.05,＜0.05,and ＜0.01,in proximal RCA,and ＜0.01,＜0.01,＞0.05,＞0.05,and ＜0.01,in LMA,respectively).Image quality was rated as excellent,general and bad for 631,330,and 37 segments in group A,640,323,and 41 segments in group B,and 615,348,and 45 segments in group C,respectively,there was no significant difference in image quality scores among 3 groups (x2 =1.90,P ＞ 0.05).Conclusion For the patient with BMI≤21.5 kg/m2,dual-source CTCA with 80 kV tube voltage setting can be used to diagnose coronary artery disease and the contrast media lose and radiation dosage can be reduced obviously.

The reverse problem of Electrical impedance tomography(EIT) is a highly ill-posed problem.It is concluded that spatial prior information could improve the final image quality.This paper proposes a new method for obtaining prior information.By this method,the inspected cross-section contour and internal structure for EIT can be achieved.

Objective To investigate the clinical effect of shuxuening treatment on organophosphorus pesti-cide poisoning with toxic myocarditis.Methods 60 patients with organophosphorus toxic myocarditis were selected in our hospital as the research subjects,and 60 cases were divided into two groups:observation group(n =30) and control group(n =30).Control group was given conventional treatment,treatment group was given shuxuening injec-tion 14 days on the basis of conventional treatment.After treatment,creatine kinase isoenzyme (CKMB),troponin (TNI),interleukin 6 (IL -6) and cholinesterase(ChE) were compared,and the changes of clinical symptoms were observed at the same time.Results There was no significant difference between the observation group and the control group(χ2 =0.630,P =0.730).The TNI,IL -6,CKMB could reflect the severity of myocardial injury in patients with different degrees of organic phosphorus poisoning,TNI,CK -MB,IL -6 increased with the degree of poisoning,the differences were statistically significant(F =11.863,4.512,3.774;P =0.000,0.015,0.029).After treatment for 4, 9,14 days,TnI,CK,CK -MB,levels of IL -6 in the two groups were recovered,but the recovery levels of TnI,CK -MB and IL -6 of the observation group significantly better than the control group,the differences were statistically sig-nificant(Fourth days,t =8.125,5.128,10.461;P =0.000,0.001,0.000;Ninth days,t =5.464,4.674,9.510;P =0.001,0.002,0.000,t =6.162,8.248,5.523;P =0.000,0.000,0.001).Conclusion Conventional treatment combined with shuxuening in the treatment of organophosphorus pesticide poisoning with toxic myocarditis has better therapeutic effect,it is worthy of promotion.

OBJECTIVE:To determine the content of Tectoridin in Rhizoma Iridis Tectori by HPLC. METHODS: The chromatographic separation was performed on Diamond ODS C18 column(150 mm?4.6 mm,5 ?m) with mobile phase consisted of MeOH-0.5% acetic acid(3∶7) at a flow rate of 0.8 mL?min-1. The detection wavelength was set at 265 nm.RESULTS: Good linear relationship was achieved over the concentration range of 0.079 2～0.396 0 ?g for tectoridin with an average recovery of 99.80%(RSD=1.09%,n=6). CONCLUSION: The method is sensitive, simple and accurate, and it can be used for the quality control of Rhizoma Iridis Tectori.