OBJECTIVE:To study the contents changes of polysaccharide and 5-HMF in Polygonati rhizoma from different pro-ducing areas after different processing,and provide reference for the development of processing technology of Polygonati rhizoma from different producing areas and the quality standard of different processing products. METHODS:UV spectrophotometry and HPLC were conducted to respectively determine the contents of polysaccharide and 5-HMF,and compare the content differences of polysaccharide and 5-HMF in Polygonati rhizoma from different producing areas [Shaanxi Lueyang County,Shaanxi Huangling County,Yunnan Fumin County (genuine producing areas),Shaanxi Taibai County] by fresh-cutting,dry-cutting,steaming and steaming with wine. RESULTS:Polysaccharide of sample from Yunnan Fumin County showed the highest content in fresh-cut sam-ples(13.4%),no 5-HMF(0)was detected;polysaccharide contents were respectively 10.8%-13.4%,8.9%-10.8%,5.5%-6.9%, 5.6%-6.5% after fresh-cut,dry-cut,steamed and steamed with wine,5-HMF contents were 0,0,0.21%-0.50%,0.25%-0.72%. Compared with no processing samples (fresh-cut),polysaccharide contents in Polygonati rhizoma were decreased in turn after dry-cut,steamed and steamed with wine,5-HMF contents were increased in turn after steamed and steamed with wine. CONCLU-SIONS:It is suggested to consider origin factor in developing processing technology of Polygonati rhizoma from genuine and non-genuine producing areas. 5-HMF content determination index should be added into quality standard of processing products after steamed and steamed with wine.

Objective To explore the detailed underlying molecular and signaling mechanisms in the effects of icariin on bone formation by an in vitro cell model. Methods The proliferation of MC3T3-E1 osteoblast-like cells was evaluated by MTT, and gene expression of cell cycle related proteins in MC3T3-E1 cells after icariin treatment was detected by real-time PCR. The phosphorylation of MAPK signals, including ERK, P38, and JNK was determined by Western blot, and then the inhibitors of MAPK signals were used to treat cells with icariin alone or together to determine the role of MAPKs in the process of icariin treatment on MC3T3-E1 cell proliferation. Alkaline phosphatase and Alizarin red staining were used to detect the formation of mineralization nodules, and gene expressions of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in osteoblasts after being treated by icariin were evaluated by real-time PCR. ICI182780, and nilutamide was used to decide the participation of estrogen and androgen receptor signals in the process of icariin treatment on the differentiation and mineralization of MC3T3-E1 cells. Results Treatment with icariin promoted MC3T3-E1 cell growth in a time- and dose-dependent manner. This treatment also revealed that icariin increased the expression of mRNAs encoding both cyclin E and PCNA, positive regulators of cell growth, but decreased levels of mRNAs encoding Cdkn2b, a negative regulator of cell cycle progression. When MC3T3-E1 cells were cultured in a differentiated condition, icariin enhanced mineralized nodule formation and increased the expression of mRNAs encoding alkaline phosphatase, type Ⅰ collagen, and osteocalcin. Treatment with icariin significantly induced phosphorylation of both ERK and JNK and this phosphorylated effect occurred very rapidly within 5 minutes and reached peak at 15 minutes. Furthermore, the stimulated effects of icariin on proliferation and gene expression of cyclin E, PCNA, and Cdkn2b in MC3T3-E1 cells were dramatically attenuated by treatment with both U0126 and SP600125, inhibitors of MAPKs. Interestingly, such stimulating effects of icariin were at least partly reduced by treatment with ICI182780, an inhibitor of estrogen receptor. Icariin induced mineralized nodule formation and gene expression of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in MC3T3-E1 cells were also partly reduced when the cells were treated with ICI182780. Conclusions Our findings indicate that the anabolic effect of icariin on bone formation is, at least partly, mediated through the MAPK signaling pathway in order to modulate osteoblast proliferation and differentiation.

A highly sensitive electrochemical deoxyribonucleic acid(DNA) biosensor based on multi-walled carbon nanotubes(MWNT)/Ag-TiO_2 composite film was developed. The solution containing Ag-TiO_2-MWNT composite was casted on the carbon paste electrode surface to form a robust film, which combine the advantages of the good biocompatibility of Ag-TiO_2 naocomposite and the fine conductivity, as well as the large active surface area of carbon nanotubes. The composite could greatly improve the immobilization capacity of the probe DNA. The morphologies and electrochemistry of the nanocomposite film were investigated by scanning electron microscopy and electrochemical techniques including electrochemical impedance spectroscopy and cyclic voltammetry, respectively. DNA hybridization events were monitored by a label-free method of electrochemical impedance spectroscopy. This label-free electrochemical impedance DNA biosensor showed high sensitivity and selectivity for phosphinothricin acetyltransferase gene sequence assay. The multicomponents films also displayed a high stability during repeated regeneration and hybridization process.

Objective To summarize our experience in the diagnosis and management of popliteal artery entrapment syndrome (PAES). Methods This is a retrospective study on 10 patients (13 limbs)who were admitted for symptoms of claudication and the diagnosis of popliteal entrapment was established either with angiography,computed tomographic angiography,magnetic resonance angiogram or during the operation in recent 7 years (2002-2009).All patients were treated surgically. Results The mean age at the time of presentation was (25 ±7) years old (range,17-41 years).Claudication was the most frequent presenting symptom (12 limbs).The surgical procedures consisted of simple musculotendinous dissociation in 1 limb,thrombectomy with balloon angioplasty in 1 limb,musculotendinous dissociation plus thromboendarterectomy with autogenous saphenous vein (ASV)patch angioplasty in 2 limbs,ASV graft interposition or bypass in 6 limbs and graft interposition or bypass in 3 limbs.At a median follow-up of (35 ±27) months (range,2 months-7 years),there were no intraoperative or long-term postoperative complications and all the patients were cured. Conclusions PAES is an unusual but important cause of peripheral vascular insufficiency especially in young patients.A combined approach is necessary for diagnosis.Popliteal artery release alone or with vein bypass or reconstruction is the treatment of choice.

Objective To investigate the clinical efficacy and indication of surgical treatment of L5S1 tuberculosis us?ing a medial edge of rectus abdominal muscle and retroperitoneal approach. Methods From Jan. 2010 to Nov. 2014, totally 18 cases of L5S1 tuberculosis were surgically cured by debridement, fusion and internal fixation via a medial edge of rectus ab?dominal muscle and retroperitoneal approach. There were 10 males and 8 females, with an average age of 35.6±10.1 years (21 to 55). The mean course of disease was 5.3±2.2 months. The primary destruction lesions were located at L5S1, and one patient of whom L4 was also involved, three cases S2 involved, and one S3 involved. Abscess pre sacrum was noticed in all cases. Six patients were combined with abscess in vertebral canal. Neural deficiency was detected in 3 patients, all grading as Frankel D. Dubousset's lumbosacral angle was 104°-126°, mean 114.6°±6.0°. Formal anti?tuberculosis medicine treatment was given for at least 2-3 weeks. All patients were treated using a medial edge of rectus abdominal muscle and retroperitoneal approach. Af?ter debridement, a tri?cortical iliac bone was used for structural inter?vertebrae fusion. The iliac bone was fixed to the sacrum by one cortical screw. Operation time, volume of blood loss, and surgery complications, including wound infection, looseness and shift of internal fixation, sinus occurrence, vascular injury, retrograde ejaculation in male patients were evaluated. The X?ray and 3D CT of lumbosacral spine, together with ESR and CRP were used to evaluate the control of tuberculosis during fol?low?up. The Dubousset's lumbosacral angle, visual analogue score, and the Frankel grade were compared between preoperative and postoperative. The bone graft fusion were also observed. Results The operation time was 147±16 min in average. The mean blood lose was 275 ± 77 ml. No vascular or neurological injury was noticed. There was also no retrograde ejaculation happened in this group. All patients were followed up from 18 to 28 months. Dubousset's lumbosacral angle was 122.4°±3.8° post?operatively and 121.4°±3.7° at the final follow?up. There was a significant difference between the lumbosacral angles pre and post operation, whereas no statistical difference between the post operation and the final follow?up. Bone fusion was noticed in all cases. All pa? tients have intact neurological function at the final follow?up. No screw breakage or graft bone migration occurred. VAS was 5.6± 1.09 pre?operation and 0.83±0.71 at the 3rd month's follow?up, and there was a significant difference between them. Conclusion Surgical treatment of L5S1 tuberculosis by debridement, fusion and internal fixation via a medial edge of rectus abdominal muscle and retroperitoneal approach is an effective method in terms of minimal trauma, low rate of modality, good ability in restore and maintenance of lumbosacral junction alignment and high fusion rate.

Objective To compare the influence for intravenous dexmedetomidine and midazolam during combined spinal and epidural anesthesia (CSEA) on sedation,respiratory and circulatory.Methods Ninety patients with lower extremity fractures and internal fixation,were divided into dexmedetomidine group,midazolam group and control group by random digits table with 30 cases each.CSEA was performed at L3-4 interspace.After block reached T8 level,dexmedetomidine,midazolam and 0.9％ sodium chloride were given to the three groups.Ramsay score,mean arterial pressure (MAP),heart rate (HR),partial pressure of carbon dioxide in end expiratory gas (PErCO2),respiratory rate (RR) were recorded before anesthesia(T0),after CSEA (T1),and 10,15,30,45,60 min after giving drug (T2-T6),and intraoperative awareness was recorded.Results Ramsay score in dexmedetomidine group and midazolam group at T2-T6 were higher than those in the group T0,T1 and concurrent control group (P ＜ 0.05),MAP were lower than those in the group T0,T1 and concurrent control group (P ＜ 0.05).HR in dexmedetomidine group at T2-T6 were lower than those in concurrent midazolam group and control group (P ＜ 0.05).PETCO2 in midazolam group at T2-T6 were higher than those in concurrent dexmedetomidine group and control group (P ＜ 0.05),RR were lower than those in concurrent dexmedetomidine group and control group (P ＜ 0.05).The rate of intraoperative awareness in dexmedetomidine group and midazolam group was lower than that in control group [16.7％(5/30) and 13.3％(4/30) vs.93.3％(28/30),P＜0.05].Conclusions Dexmedetomidine and midazolam provide good sedation to reduce intraoperative awareness,slight inhibition of blood pressure.Dexmedetomidine can decrease HR,but it does not influence respiratory function.Midazolam restrains respiratory function.

Objective To investigate demography of sensory tricks in cervical dystonia (CD),and to discuss the relationship between sensory tricks and the function of sensorimotor integration of cerebral cortex,meanwhile to explore potential worthiness using sensory tricks in the management of CD.Methods We recruited 75 patients (23 male,52 female,male female ratio:1:2.26;aged 19-72 years,mean (42.45 ± 13.09) years) admitted to the Dystonia Clinic Center of the First Affiliated Hospital of Dalian Medical University from March 2016 to April 2013.Standardized questionnaire on sensory tricks was used to investigate the sensory tricks first recognized in 75 patients retrospectively.Median nerve somatosensory evoked potential via multipoint recording was measured in 43 CD patients who manifested as simple rotation of neck.Bilateral amplitudes of P22/N30 derived from F (3,4) were compared.Results Frequency of sensory tricks in CD was 85.53％ (64/75) in our research.As disease progressed,the type of sensory tricks used by patients may transform from one to another or even disappear.No statistically significant differences were found in ages among patients using classic sensory tricks,forcible tricks and patients without sensory tricks,whereas the complexity of the disease showed increased tendency among the three groups.Bilateral P22/N30 amplitudes of patients with classic sensory tricks showed no significant differences.P22/N30 amplitudes contralateral to the head turn were significantly higher than ipsilateral in patients using forcible tricks ((3.16 ± 1.71) μV vs (2.47 ± 1.28) μV,t =2.243,P=0.038) and patients without sensory tricks ((3.62 ±1.58) μV vs (2.73 ±1.14) μV,t=2.893,P=0.023).Tsui scores among patients using classic sensory tricks (9.36 ± 3.52) and forcible tricks (12.67 ± 5.00) or patients without sensory tricks (15.18 ± 4.07) had statistically significant differences (classic sensory tricks vs forcible tricks,t =-3.020,P =0.004;classic sensory tricks vs patients without sensory tricks,t =-4.452,P =0.000).Conclusions Types of sensory tricks used by patients are associated with severity of the disease.Different types of sensory tricks may reflect status of adaptive compensatory mechanism of the cerebral cortex.

UNLABELLED: Abstract OBJECTIVE: To measure the expression of CK18 and CK19 in the cells from peripheral blood and tumor tissue of the nasopharyngeal carcinoma patients,to test whether CK 18 and CK 19 could be biomarkers of nasopharyngeal carcinoma fordiagnosis. METHOD: The mRNA was extracted from the blood and carcinoma tissue of nasopharyngeal carcinoma and was reversed transcription to cDNA. The 3 pairs primers were designed for RT-PCR and the fold value was calculated to evaluated expression by ΔCT. RESULT: There are no statistical differences between the CK18 and CK19 gene expression and the gender, age and metastasis in tumor tissue of 45 nasopharyngeal carcinoma patients (P>0. 05). There are significant differences among 3 pathological stages and 2 genes expressed increase as the grade malignancy (P<0. 05). The detecting of the 2 genes expression from blood cells shows that CK18 and CK19 had a high positive ratio 64% and 75% respectively. Meanwhile this method showed a same detection characteristic in tumor and blood, the positive.rate of CK18 and CK19 genes in metastasis is higher than non-metastasis. The results showed CK18 has a high specificity and CK19 has a high sensitivity for prognosis and all relapsed cases are associated with the expression of CK18 and CK19. CONCLUSION CK18 and CK19 may be used as biomarkers of nasopharyngeal carcinoma for diagnosis.
Biomarkers, Tumor ; Carcinoma ; DNA, Complementary ; Gene Expression ; Humans ; Keratin-18 ; biosynthesis ; Keratin-19 ; biosynthesis ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; diagnosis ; metabolism ; pathology ; Neoplasm Metastasis ; Prognosis ; RNA, Messenger ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the effects of icarrin on the activity and protein expression of core binding factor otl(Cbfa1) in rat osteoblasts cultured in vitro,and to explore whether mitogen-activated protein kinase (MAPK) pathway is involved in this process.Methods Calvarial osteoblasts were obtained from newborn (<24 h) SD rats by trypsin-coUagenase digestion method.The second generation osteoblasts were cultured in the medium containing icariin (10 ng/ml) or estradiol (10-8 mol/L) with or without extracellular-signal regulated kinase (ERK) inhibitor (UO126) or p38MAPK inhibitor (SB203580).Nuclear protein was extracted from osteoblasts.And then the activity of Cbfa1 was detected by ELISA.The amounts of Cbfa1 protein were detected by Western blot.Results Calvarial osteoblasts were obtained successfully and were used in this study after indentified by alkaline phosphatase and mineralized nodus staining.Cbfa1 expression and the activity in osteoblasts were up-regulated by both icariin and estradiol (P<0.05).The effects were partly inhibited by addition of U0126or SB203580 (P<0.05).Conclusions Either icarrin or estradiol can stimulate the proliferation and maturation of cultured osteoblasts in vitro via up-regulating the activity and expression of Cbfal.The MAPK signal pathway inhibitor seems to partly decrease Cbfa1 activity.It suggests that MAPK pathway may be involved in the transduction of icariin's impact on proliferation and mineralization of osteoblasts.

Objective To study the effect of peroxisome proliferator-activated receptor garama (PPARγ) ligand on hepatic fibrosis in rats. Methods Forty Wistar rats were randomly divided into two groups: the control group(20 rats) ,in which liver cirrhosis was induced by adding 0. 3‰ thioacetamide in the fodder for 6 months, and rosiglitazone group(20 rats) in which 200 ppm of rosiglitazone in combination with 0. 3‰ thioacetamide was added in the fodder. Liver tissue's mRNA expression of PPARγ, TGF-β1 ,type Ⅰ pro-collagen and α-smooth muscle actin(α-SMA) was detected by RT-PCR. The protein expression of PPARγ, TGF-β1 ,type Ⅰ collagen and α-SMA was detected by Western blot. The expression of collagenof liver histological section was evaluated by Van Gieson (VG) staining. Results The expression ofPPARγ at mRNA level significantly increased in rosiglitazone group compared with those in the control group ( t = 6. 93, P < 0. 01 ), while the expression of TGF-β1 ( t = 3. 89, P < 0. 01 ) and type Ⅰ pro-collagen ( t =5.67,P <0. 01 ) were lower than that in the control group. The protein expression of PPARγ, TGF-β1 and type Ⅰ collagen was in similar tendence with that of mRNA expression. The expression of α-SMA decreased significantly in rosiglitazone group compared with that in the control group (t = 3. 12,P < 0.01 ). The collagen stainings of liver histological section in rosiglitazone group was lower than those in the control group (t = 3.47, P < 0. 01 ). Conclusion PPARγ ligand inhibits the production of collagen in fibrofic livers in rats and prevents hepatic fibrosis in vivo.