Objective To evaluate the feasibilty of modified osteogenic culturing of goat adipose derived stromal cells(ADSCs).Methods From March,2013 to September,2014,platelet-rich plasma(PRP) was made from goat autogenous vein blood,and abodimoneal fat was aspirated,following aspetic procedure,primary and series passage of ADSCs was established.Osteoinduced ADSCs was carried out according to following group:combinative osteoinduction group(PRP+rhBMP-2 +ADSCs),growth factor group(rhBMP-2+ADSCs),conventional inductive group(dexamathasone+ascorbic acid + ADSCs) and noninductive group(blank group).Converted microscope was used to observe cellular pattern,cell activity,osteocalcin and collagen type Ⅰ level were detected at 1,3,5,7,9,13,17,21 days.Red Alazarin and Von Kossa staining were also assayed at different interval.Results Under observation of converted microscopy,remarkable cell proliferation with abundant ECM was noticed in combinative osteoinduction group,compared with other groups,level of cell activity,osteocalcin,collagen type Ⅰ [(0.82 ± 2.19)AU,(79.82 ± 1.36)U/L at,(78.51 ± 4.32)μ g/ml at]were significantly higher than other groups (P ＜0.05),and remarkable ALP expression and calcifed nodules were also seen.Conclusion PRP can enhance the inductive effect of rhBMP-2,and combinative osteoinduction procedure acts as a satisfactory culturing method.

Objective To evaluate cardiac biomarkers as biological risk factors for cardiovascular and all-cause motality in HD patients.In addition,a multimarker approach including inflammatory index was performed to improve the cardiovascular and all-canse risk assessment of these patients.Methods The author measured Troponin-T (TnT),N-terminal pro brain natriuretic peptide (NT-proBNP) and high-sensitivity C-reactive protein (HsCRP),collected the clinical data at baseline (January 2012) in 229 HD patients in three hemodialysis centers in Haidian District of Beijing,recorded time and cause of death in the next 1000 days.Kaplan-Meier was used to calculate survival rate and impact factors of prognosis.Cox proportional hazard model was used to estimate significance of =TnT,NT-proBNP and HsCRP and adjusted hazard ratios (HRs) of death.Results During the followup,37 patients died,mainly from cardiac cause (54.05％,20/37).Univariate analysis found old age,diabetes,cardiovascular disease,low serum albumin,CRP≥3 mg/L,TnT≥0.1 mg/L,NT-proBNP≥ 4381 ng/L were associated with prognosis.Elevated cTnT,NT-proBNP or HsCRP were all associated with increased cardiovascular and all-cause motality.Moreover,the combination of all parameters (NTproBNP≥4381 ng/L and TnT≥0.1 mg/L and HsCRP≥3 mg/L) were dramatically associated with increased cardiovascular cause mortality (HR=25.25,P ＜ 0.01) and all-cause mortality (HR=27.33,P ＜ 0.01).The association were significant even after full adjustment for cardiovascular (HR=14.33,P ＜ 0.01) and all-cause mortality (HR=1 1.54,P ＜ 0.01) respectively.Conclusions A combined index of cardiovascular risk factors could provide supplementary risk stratification in HD patients for cardiovascular mortality and all-cause mortality,strongly supporting the annual routine determination of these biomarkers.

Aims To observe the effect of saccharide extracts of Yiqijianpi herb Codonopsis and Glycyrrhizae on polyamine-dependent activation of K+ channels sig-nal pathway during cell migration and to investigate their mechanism of promoting restoration in gastrointes-tinal mucosal injuries. Method The study was based on IEC-6 cell migration model. While in a normal polyamine level or polyamine was inhibited by DFMO, the effect of Codonopsis saccharide extracts and Glycyr-rhizae saccharide extracts on polyamine-dependent acti-vation of K+ channels signal pathway during cell mi-gration was observed. (1) K+ channel protein Kv1. 1 was determined by Western blot. (2)Membrane poten-tial was measured by Flow Cytometer. (3) Laser scan-ning confocal microscope was used for measuring [ Ca2+] cyt. ( 4 ) The expression of RhoA, which is Ca2+ downstream protein, was determined by Western blot. Results During cell migration, Codonopsis and Glycyrrhizae saccharide extracts could: ( 1 ) improve the expression of Kv1 . 1 protein and ameliorate the de-crease of kv1. 1 protein expression by DFMO;(2) in-crease membrane hyperpolarization and reverse mem-brane depolarization resulted by DFMO; ( 3 ) improve intracellular [ Ca2+] cyt, while Codonopsis could re-verse the decrease of [ Ca2+] cyt caused by DFMO;(4) improve the expression of RhoA protein, reversing its decline caused by DFMO. Conclusion Codonop-sis and Glycyrrhizae saccharide extracts can promote cell migration in IEC-6 cell, which is correlated with their effect on polyamine-dependent activation of K+channels signal pathway.

OBJECTIVE To observe the effect of Codonopsis and Glycyrrhiza glycoconjugates on migration and membrane potential of small intestinal epithelial cells(IEC-6),and to explore the promoting effects of Yiqi jianpi herb Codonopsis and Glycyrrhiza on gastrointestinal mucosal injury repair and the underlying mechanisms. METHODS Under normal conditions or loaded with the inhibitor of potassium channel 4-aminopyridine(4-AP),IEC-6 cells were treated with Codonopsis and Glycyrrhiza glycoconjugates (25-200 mg · L-1) for 24 h,respectively. IEC-6 cell migration was observed by the phase contrast microscope and cell membrane potential was detected by flow cytometry. RESULTS Codonopsis and Glycyrrhiza glycoconjugates (50 and 100 mg · L-1) increased the number of migrated IEC-6 cell compared with normal control group(P<0.01,P<0.05). Compared with normal control group,4-AP reduced the number of migrated IEC-6 cell(P<0.01). Codonopsis and Glycyrrhiza glycoconjugates (50-200 mg · L-1)reversed cell migration inhibited by 4-AP significantly when compared with 4-AP model group(P<0.01). The results of flow cyometry analysis showed that the cell membrane poten?tial was increased after treatment with Codonopsis and Glycyrrhiza glycoconjugates(50 mg · L-1)compared with normal control group and resulted in an increase in cell membrane hyperpolarization(P<0.01). Compared with normal control group,4-AP decreased the cell membrane potential(P<0.01)and resulted in cell membrane depolarization. Also,compared with 4-AP model group,cell membrane depolarization induced by 4-AP was reversed by treatment with Codonopsis and Glycyrrhiza glycoconjugates(100 and 200 mg·L-1). CONCLUSION Codonopsis and Glycyrrhiza glycoconjugates may promote gastrointestinal mucosal injury repair and the mechanisms may involve the activation of signaling pathways by affecting polyamine-dependent intestinal epithelial cell migration voltage-gated K+channels.

Objective To analyze the effect of interventional treatment in patients with swelling hand syndrome.Methods 100 patients with maintained hemodialysis were enrolled in this study (49 male,51 female).Upper limb swelling involved the dialysis was examined with a course of 1 week to 36 months.Before intervention treatment,stenosis or occlusion of the brachiocephalic vein or subclavian vein were found in 95 patients diagnosed by CTA.Angiography was performed through the affected side of the upper limb.Then a balloon pre-dilatation of the stenosis or occlusion was performed,followed by balloon dilatation or stent molding treatment through the femoral vein.Results Percutaneous transluminal angioplasty or stent angioplasty was performed in 87 patients without severe complications,while treatment was failed in 13 cases.Swelling was gone in 85 patients 2-5 days after surgery,and persistent symptom was observed in 2 cases.All the 87 patients were followed up from 2 to 46 months after interventional treatment,one patient died of cancer after 14 months.Relapsed symptom of swelling hands was found in 20 patients and interventional therapy was performed again.Conclusion Swelling hand syndrome is common in maintained hemodialysis patients due to the stenosis or occlusion of central vein involved the dialysis side.Interventional treatment is a safe and effective procedure.

Abstract Dry eye is a multifactorial ocular surface disease characterized by tear film homeostasis imbalance, which usually causes eye discomfort and visual impairment. With the change of environment and lifestyle, the incidence of dry eye in children and adolescents has risen steadily. Epidemiological studies have confirmed that the incidence of dry eye is closely related to unhealthy lifestyle. Prevention should be given great priority to in avoiding lifestyle related dry eye. The review summarizes the definition, epidemiology, risk factors and preventive measures of lifestyle related dry eye, so as to provide a theoretical basis for improving the diagnosis, treatment and prevention of lifestyle related dry eye in children and adolescents.

Primary bone lymphoma(PBL)is rare and has no specific clinical features.A mouth-eaten pattern of bone destruction in metaphysic or subchondral bone and a large soft-tissue mass with little cortical destruction are important imaging features of primary bone lymphoma. The final diagnosis mainly depends on the histopathological features and the expression of specific proteins showing by immunohistochemical staining. It bears a good outcome after radiation and chemical therapy. The operation is not a routine therapy method.So, it is important to make accurate diagnosis before radiation and chemical therapy. In this article, we reviewed the clinical diagnosis, treatment, and prognosis of primary bone lymphoma.

Corpus Callosum infarction is relatively rare in clinical practice, of which the pathogenesis is still unclear. The artical reports a patient with bilateral corpus callosum infarction associated with Moyamoya syndrome, discussing the etiology from the perspective of vascular anatomy and risk factors. With the review of relevant clinical cases home and abroad nearly a decade, we analyzed and summarized the vascular features of corpus callosum infarction and the main etiology, in order to contribute to clinical diagnosis and treatment.

Objective: To evaluate the role of neuroligin 1 (NL-1) in trafficking of GluR1-containing AMPA receptor to cell membrane in spinal cord dorsal horns during remifentanil-induced hyperalgesia in mice with incisional pain. Methods: Forty SPF healthy male C57BL/6J mice, aged 8-10 weeks, weighing 18-22 g, were divided into 5 groups (n=8 each) using a random number table method: control group (group C), NL1-shRNA plasmid group (group NL), incisional pain plus remifentanil group (group I+ R), incisional pain plus remifentanil plus blank vector group (group I+ R+ B), and incisional pain plus remifentanil plus NL-1-shRNA plasmid group (group I+ R+ NL). Negative lentivirus was intrathecally injected in group I+ R+ B.In NL and I+ R+ NL groups, 10 μl NL-1-shRNA lentivirus at 1×10⁸ IFU/ml was intrathecally injected once a day for 3 consecutive days.Normal saline 10 μl was intrathecally injected at the same time point in C and I+ R groups.After transfection was stable, normal saline 0.1 ml was injected via the caudal vein for 4 consecutive times at 15 min intervals in C and NL groups.In I+ R, I+ R+ B and I+ R+ NL groups, 0.1 ml remifentanil 10 μg/kg was injected via the caudal vein for 4 consecutive times at 15 min intervals, and the model of incisional pain was established after the first administration.The mechanical paw withdrawal threshold (MWT) and tail-flick latency (TFL) were measured at 24 h before normal saline or remifentanil administration (T₀) and at 3, 6, 24 and 48 h after the end of administration (T_1-4). The animals were sacrificed after measurement of pain threshold at T₄, and L_4-6 segments of the spinal dorsal horn were then collected for determination of the expression of NL-1 protein and mRNA and AMPA receptors, and the ratio of AMPA receptor expression in the membrane protein to that in the total protein (m/t ratio) was calculated. Results: Compared with group C, the MWT was significantly decreased, and TFL was shortened at T_1-4, the expression of NL-1 protein and mRNA and GluR1-containing AMPA receptors in membrane and total proteins was up-regulated, and m/t ratio was increased in I+ R and I+ R+ B groups (P<0.05). Compared with I+ R and I+ R+ B groups, the MWT was significantly increased and TFL was prolonged at T_1-4, the expression of NL-1 protein and mRNA and GluR1-containing AMPA receptors in membrane and total proteins was down-regulated, and m/t ratio was decreased in group I+ R+ NL (P<0.05). Conclusion NL-1 in spinal cord dorsal horns can promote the trafficking of GluR1-containing AMPA receptors to cell membrane, which is involved in the development and maintenance of remifentanil-induced hyperalgesia in mice with incisional pain.

Objective: To evaluate the role of spinal COX-1 and COX-2 in remifentanil-induced hyperalgesia in mice with incisional pain. Methods: Thirty-two male C57BL/6J mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups (n=8 each) using a random number table method: control group (group C), incisional pain plus remifentanil group (group IR), incisional pain plus remifentanil plus selective COX-1 inhibitor group (group IR+ SC560), and incisional pain plus remifentanil plus selective COX-2 inhibitor group (group IR+ SC236). In IR, IR+ SC560 and IR+ SC236 groups, normal saline 10 μl, SC560 25 μg and SC236 25 μg were intrathecally injected, respectively, 15 min later remifentanil 10 μg/kg was injected via the tail vein for 4 times at 15 min intervals.An incisional pain model was established after the first injection of remifentanil.The mechanical paw withdrawal threshold (MWT) was measured at 24 h before normal saline or remifentanil injection and 3, 6, 24 and 48 h after the last injection (T₀-T₄). The mice were sacrificed after the last measurement of pain threshold, and the L_4-6 segments of the spinal cord were removed for determination of the expression of COX-1 and COX-2 (by Western blot) and expression of COX-1 and COX-2 mRNA (by quantitative real-time polymerase chain reaction). Results: Compared with group C, the MWT was significantly decreased, and the expression of COX-2 protein and mRNA was up-regulated in IR, IR+ SC560 and IR+ SC236 groups (P<0.05). Compared with group IR, the MWT was significantly increased in IR+ SC560 and IR+ SC236 groups (P<0.05). There was no significant difference in the MWT at each time point between IR+ SC560 and IR+ SC236 (P>0.05) .There was no significant difference in the expression of COX-2 protein and mRNA among group IR, group IR+ SC560 and group IR+ SC236 (P>0.05). There was no significant difference in the expression of COX-1 protein and mRNA among the four groups (P>0.05). Conclusion Compared with COX-1, spinal COX-2 plays a major role in the pathophysiological mechanism of remifentanil-induced hyperalgesia in mice with incisional pain.