OBJECTIVEThis study aims to explore the effect of Notch signaling depression on the receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis of RAW264.7 cells.

METHODSMice RAW264.7 cells were cultured and differentiated into osteoclasts with the induction of RANKL. The expressions of Notch1, Notch2, Deltal, Jagged1, Hes1, tartrate-resistant acid phosphatase (TRAP), and Cathepsin K genes during osteoclastogenesis were analyzed using real-time polymerase chain reaction. Osteoclast formation was analyzed using TRAP assay with suppression of Notch receptors by a selective γ-secretase inhibitor (GSI).

RESULTSNotch1, Notch2, Delta1, Jagged1, and Hes1 expressions in RAW264.7 cells were upregulated following 50 ng · mL-RANKL stimulation for 3 d, concomitant with the expression of the osteoclast differentiation markers TRAP and Cathepsin K. Notch2 and Jagged1 had the most remarkable increase in the Notch family members. GSI inhibited RANKL-induced osteoclastogenesis of RAW264.7 cells and Hes1 expression dose-dependently.

CONCLUSIONNotch signaling activation may promote RANKL-induced osteoclastogenesis of RAW264.7 cells.

Adult ; Aged ; Asteraceae ; Drugs, Chinese Herbal ; adverse effects ; Female ; Hepatic Veno-Occlusive Disease ; chemically induced ; Humans ; Male ; Middle Aged ; Retrospective Studies

BACKGROUND: Techniques in the focal cerebral ischemia model in mice have been well developed in some foreign countries. However, rats are commonly used in cerebral ischemia studies due to resource availability in China.OBJECTIVE: To establish reliable and reproducible focal cerebral ischemia reperfusion model in mouse to explore the molecular mechanism of cerebral ischemia reperfusion process in genetic level.DESIGN: Randomized controlled study.SETTING: Fundamental Medical Research Institute of Chengde Medical College. MATERIALS: The experiment was carried out in Fundamental Medical Research Institute of Chengde Medical College from September 2002 to May 2003. Totally 30 Kunming mice (25 to 30 g), provided by Experimental Animal Center, Chengde Medical College, were randomly grouped as follows: control group, 6, 3-hour occlusion and 18-hour, 24-hour reperfusion, with 6 mice in each group respectively.INTERVENTIONS: Nylon thread (qb0.128 mm) was soaked in paraffin wax and inserted into internal carotid artery in mouse to reversibly occlude the middle cerebral at the right side after 3 hours, then was removed, and reperfusion was performed in 18, 24 hours, respectively.MAIN OUTCOME MEASURES: The infarct volume was observed and neurological deficit scored was analysis.of mice in operation group was shown as head turning to left, adduction of left forelimb, internal rotation of shoulder, decrease of muscular tension of left forelimb, unable to straighten completely during tail lifted. At independent activity, mice turned to left as the left hidlegs as the center of a circle; under the abdomen, left forelimb was crossed with right forelimb as scissor shape. Palsy of left limbs was observed obviously, especially forelimb.tion was shown clearly with TTC. The infarct volumes were (13±2.3) mm3 and (16±4.5) mm3 for 18 hours and 24 hours reperfusion and 3-hour ischemia, respectively. Pathological changes were observed under light microscope after ischemia. The infarct volume increased as the increasing of reperfusion time, including 3-hour blocking and 18-hour and 24-hour reperfusion.CONCLUSION: Using suture method to establish focal cerebral ischemia reperfusion model in mice, it is less invasion and trauma without requiring skull opening. Ischemia position is relatively fixed, thus, ischemia reperfusion time can be controlled precisely. It is also an ideal animal model to study pathological changes in cerebral vascular diseases and an efficient tool to study the therapeutic effect of medicine after surgery.

Objective To discuss early and standard treatment of open calcaneal fractures so as to lower incidence of amputation and osteomyelitis. Methods From October 2005 to October 2006,16 cases of 17 open calcaneal fractures were treated in our department.There were 12 males(13 fractures)and 4 females at a mean age of 31 years.All cases were treated with the sanle early treatment protocol,including emergent debridement,and lavaging with normal saline,H2O2 and iodide solutions.All cases were immobilized with plaster or multiple K-wires according to systemic condition and fracture type.With detumescence of the soft tissues.open reduction and internal fixation was done via lateral incision.The patients were followed up for mean six months and the incidence rate of complications evaluated bv American Orthopaedic Foot and Ankle Society(AOFAS)scale. Resuits No amputation was found.but there was one case with osteomyelitis.one with superficial infection and one with deep infection.Delayed skin union was found in one case and plantar skin necrosis in one. Conclusion Early and standard treatment of open calcaneal fractures can minimize the infection,prevent osteomyelitis and provide sound soft tissue for secondary surgical management.

Objective To investigate the effects of BKCa channel on electrophysiology excitatory regulation in MVN neuron following hypoxia and to reveal its molecular mechanism.Methods C57BL/6 mices were performed MVNs hypoxia mice model,and randomly allowed to normal oxgen group and hypoxia group.The hypoxia group, according to the application of NS1 6 1 9 ,was further divided into the no NS1 6 1 9 pretreatment group and NS1 6 1 9 pre-treatment group.Using the patch clamp experiment technology,we recorded the effects of the MVN abnormal neu-ronal firing and the change of the BKCa currents.Using immunohistochemical technique,the changes of BKCa in the hypoxic MVNs detected were.Results Acute hypoxia increased neuronal activities.NS1619 pretreatment de-creased hypoxia-induced firing rate,and increased and postponed the maximum increase by hypoxia(P<0.05),al-so alleviated 10-min-hypoxia-induced depolarization(P<0.05).Perfusion with hypoxic significantly reduced the BKCa positive neurons(P<0.05).Conclusion These findings suggest that acute hypoxia increases neuronal activi-ties.The decreased MVN BKCa channels contribute to hypoxia-induced abnormal neuronal activities.

Objective To explore the short-term clinical outcomes of ankle arthrodesis for the treatment of posttraumatic ankle arthritis.Methods Retrospectively study the data of 47 patients with posttraumatic ankle arthritis who were followed up after receiving ankle arthrodesis from January 2008 to May 2012.There were 28 males and 19 females,with an average age of 44 years (range,14-72 years).All of them wereclassified into stage-3 arthritis according to Morrey-Wiedeman,among whom 31 had obvious foot varus deformity or equines deformity,and 16 were almost normal in appearance.The ankle joint function of patients were assessed by Kofoed,American Orthopaedic Foot and Ankle Society (AOFAS) ankle and hindfoot score,Maryland foot score systems and visual analog scale (VAS) preoperatively and at the last follow-up.Results The patients were followed up from 5 to 52 months (average,25 months).All the ankles got fused at the last follow-up,with one receiving removal of internal fixation and debride because of infection.Forty-three patients were satisfied with or without reservations; the others thought it so.The satisfaction rate was 91.5％(43/47).The Kofoed,AOFAS,Maryland foot scores and VAS were significantly improved from 48.11 ±5.00,53.38±5.52,49.74±5.71,7.89±0.87 to 80.96±6.07,82.21±4.26,88.94±5.14,2.28±1.10 respectively at the last follow-up.The normotopia X-ray of ankle joint showed that there were 22 cases in which the ankle got fused at neutral position,19 cases in which the eversion was less than 5°,3 cases in which the eversion was between 5° to 10°,2 cases in which the varus was less than 5°,and 1 case in which the varus was between 5° to 10°.Lateral X-ray showed that there were 32 cases in which the ankle got fused in dorsiflexion neutral position,12 cases plantar flexion 5°,and 3 cases plantar flexion 5° to 10°.Conclusion Ankle arthrodesis is still the preferable surgical treatnent option in patients with end-stage ankle arthritis,which shows high reliability and effectively in correcting deformity,pain relieving and function improvement.

Objective We conducted a systematic review and meta-analysis of the published literature to evaluate the efficacy of prophylactic antibiotics in the prevention of incisional infection of inguinal tension-free hernioplasty.Methods Articles of randomized controlled trials about the efficacy of prophylactic antibiotics in the prevention of incisional infection in the inguinal tension-free hernioplasty published from January 1975 to October 2012 was retrieved and systematically reviewed.Results A total of 11 randomized controlled trials meeting the inclusion criteria were screened.Among the 4159 cases of patients,130 cases had wound infection,and the infection rate was 3.13％.In the prophylactic antibiotics group of 1845 cases,wound infection occurred in 45 cases; the infection rate was 1.94％.In the control group of 2314 cases,85 cases had incision infection; the infection rate was 3.67％.The preventive use of antimicrobial drugs reduced surgical site infection of inguinal tension-free hernioplasty (OR 0.55,95％ CI 0.38-0.80,P =0.002).Conclusions The use of prophylactic antibiotics can effectively reduce the incisional infection in the inguinal tension-free hernioplasty.

AIM:To investigate the changes of visual development produced by monocular atropinization in rats.
METHODS: Twenty normal SD rats were randomly divided into two groups: control group ( n = 10 ) and atropinization ( experimental) group ( n=10 ) . All the left eyes were selected as the experimental eyes, and the right eyes served as the normal eyes. The left eyes in atropinization group was produced by 1% atropine, 3 times a day and the right eyes in control group was treated with normal saline, 3 times a day. The flash visual evoked potentials ( F-VEP ) and retinoscopy refraction of the rats'both eyes were detected at five time points:0, 7, 14, 21, and 28d after atropinization, respectively. After 28d, six rats were randomly selected from both groups and each group had three rats. The expression of the c- fos mRNA was observed in both visual cortexes. Another six rats were chosen for the same test after 2d dark environment with 2h light later. The expression of c-fos mRNA was detected again.
RESULTS: After 14d anisometropia was observed in experimental group, the difference was 3. 9D ( P<
0.0 5 ) , F-VEP P1 wave of the rats left in experimental group was reached to 88. 9±1. 889ms at 21d, there was statistical difference compared with the right eye ( P<0.05). After 28d, c-fos mRNA expression in the left visual cortex of rats in the experimental group was higher than that of the right side, but there was no significant difference. But when underwent 2h light stimulation after in the darkroom 2d, the c-fos mRNA expression in in the left visual cortex of rats in the experimental group was 5 times higher than that of the right side, there was a statistically significant difference (P<0. 05).
CONCLUSION: In the critical period of visual development, monocular chronic atropine in rats can form anisometropia, may delay the transmission of the optic nerve, hinder the normal development of the visual cortex. Monocular atropinization in rats can be used as the model of anisometropia.

Objective To discuss the significance of the application of real-time quantitative PCR(RQ PCR)for diagnosis and guidance therapy of cytomegalovirus(CMV)infection after allo- hematopoietic stem cell transplantation(allo-HSCT).Methods Thirty-three patients were undergo- ing allo-HSCT.After hematopoietic reconstitution,patients'peripheral blood samples were detected for CMV DNA by RQ-PCR periodically.Anti-viral therapy was begun,attenuated and stopped ac- cording to the results of CMV DNA detection.The effects of anti-viral therapy and patients' clinical results were observed.Results CMV DNA was detected in blood samples from 13 of 33 patients. There were 21 episodes in these patients.Only 1 episode wasn't controlled hecause the patient gave up the therapy.CMV DNA copies were disappeared soon or decreased and then disappeared during anti-viral therapy in the others'.Patients which had symptoms and/or dysfunction of organs were cured too.Each course of anti-viral therapy was shorter than ordinary course.Conclusions CMV in- fection can he diagnosed as early as possible by RQ PCR.To begin,attenuate and stop anti-viral ther- apy according to the results of RQ-PCR is safe.The course is shortened and the side effects of anti vi- ral drugs were attenuated.

Objective: To establish a method to determine 4 active ingredients in Jindan tablets by HPLC.
Methods: Agilent Eclipse X DB C₁₈ (4.6 mm×250 mm, 5 μm) column was adoped using acetonitrile (phase A) -0.1% phophoric acid solution (phase B) as the mobile phase with gradient program at the flow rate of 1.0 mL·min^-1, the detection wavelength was 270 nm, Column temperature was set at 30 ℃.
Results: The linear ranges of gentiopicrin, polydatin, quercetin and emodin were 7.875-78.75 μg·mL^-1（r=0.999 9）, 6.75-67.50 μg·mL^-1（r=0.999 7）, 7.726-77.26 μg·mL^-1（r=0.999 4）, 3.809-38.09 μg·mL^-1（r=0.999 8), respectively, the peak area had a good linear relationship with the mass concentration of 4 components, the average recovery were 99.31%, 99.21%, 99.04%, 99.59%, (n=6) respectively, and RSDs were 1.86%, 1.24%, 1.37%, 1.15%, respectively.
Conclusion: This method is reliable, repeatable and stable, and has strong specificity. This method can provide a reference for the quality control and standard improvement of Jindan tablets.