Objective:To analyze the chemical constituents of Rhizoma belamcandae by using high-performance liquid chromatography-diode array detector/electrospray ionization-mass spectrometry (HPLC-DAD/ESI-MS).Methods: Dried Belamcanda chinensis powder was extracted with 70% ethanol by sonication.The chromatographic separation was performed on a YMC ODS-C18 column (250 mm?4.6 mm I.D.,5 ?m) with a mobile phase composed of 20%CH3OH(A)-70%ACN(B) (0→30→45→65 min,20→30→90→100 B),eluted at a flow rate of 0.45 ml/min,and the UV detection wavelength was set at 265 nm.Positive ionization mode with a needle voltage of 5 000 V,a capillary voltage of 20 V,a gas(N2)press of 20 psi and a temperature of the drying gas of 300℃ was selected.Relative molecular mass data acquisition was performed from m/z 250 to 550 in full MS scan mode.Results: Nine major isoflavones were identified from Rhizoma belamcandae based on their retention behavior obtained on-line by their UV spectra and the HPLC-DAD/ESI-MS.Conclusion: A rapid and efficient HPLC-DAD/ESI-MS method for identifying the chemical constituents of Belamcandae chinensis has been established,which provides more scientific information for quality control of Rhizoma belamcandae.

Objective To observe the role of baicalin on the expression of phosphorylated protein of signal transduc-ers and activators of transcription signaling proteins (STATs) during the process that neural stem cells (NSCs) differentiating into neurons. Methods NSCs were isolated from the embryonic cerebral cortex of the 14-15-day pregnant SD rats, which were cultured and passaged in vitro. The 3rd generation of NSCs was used in the experiment. NSCs were randomized into nat-ural differentiation control group, three different doses of baicalin groups (7.5μmol/L, 15μmol/L and 30μmol/L), leukemia inhibitory factor (LIF)+basic fibroblast growth factor (bFGF) group and baicalin+LIF+bFGF group. After 6 d culture in vi-tro, the immunohistochemical method was used to observe the expressions of microtubule-associated protein 2(MAP-2) and glial fibrillary acidic protein (GFAP) in different groups. The expression levels of phosphorylation protein of STAT 3 in NSCs were detected by Western blotting method after 2 h and 6 d of culture. Results The expression of MAP-2 in NSCs was in-creased by baicalin, but the expression of GFAP in NSCs decreased. The expression of GFAP in NSCs was enhanced in LIF+bFGF group, which was inhibited by baicalin+LIF+bFGF. The phosphorylation level of STAT3 in NSCs was downregulat-ed by baicalin, but the phosphorylation level of STAT3 was upregulated in LIF+bFGF group. The upregulated phosphoryla-tion level of STAT3 was inhibited in baicalin+LIF+bFGF group(P＜0.05). Conclusion Baicalin can induce NSCs to dif-ferentiate into neurons, which may be caused by the downregulation of the phosphorylation level of STAT3 in NSCs.

Flocculation of fermented broth with 1,3 -propanediol using component B of natural clarifier-Ⅱ was studied. Firstly, single factor tests and orthogonal tests were carried out. The results showed the main factors which influenced the flocculation and the optimal conditions were: pH6.0, flocculant B 0.01g/L , NaCl 0g/L, and the FR was 95.97% . The following results by filtration experiments, compared with the sample without pretreatment, indicated that the filtration speed of the flocculated sample was improved dramatically, and the weights of net and dry filter cakes were increased by 41.13% and 51.88% , respectively. Electrodialysis experiments showed that flocculation could accelerate desalting process, and could take the place of centrifugal pretreatment when it combined with the filtration.

Objective To explore the difference of DNA methylation levels between normal Schwann cells (NSCs) and activated Schwann cells (ASCs) in rats. Methods The adult Wistar rats were received sciatic nerve ligation and fed for 7 days. The ASCs and NSCs were separated from ligated sciatic nerves and brachial plexus respectively. Immunocytochemical staining of S-100 antibody was used to identify the cells. The growth condition of cells was detected by CCK-8 method. Methylated DNA immunoprecipitation sequencing (MeDIP-Seq) was applied to filter the differentially methylated regions in ASCs and NSCs. The distribution of differentially methylated genes related with axonal regeneration in chromosome was analyzed, and Gene ontology(GO)and PATHWAY analysis were also conducted. Results High purity of ASCs and NSCs were obtained successfully, which were both positive for S-100 antibody. In the same culture condition, ASCs showed a faster proliferation than that of NSCs. A total of 177 176 differentially methylated regions were found by MeDIP-Seq. Among them, 1 097 were located in the promoter (≤1 kb), 1 136 in the promoter (1-2 kb) and 567 on the CpG. After functional annotation of differentially methylated genes, 214 differentially methylated genes related with axonal regeneration were found in ASCs and NSCs. Compared with NSCs, 191 genes were up-regulated and 23 genes were down-regulated in ASCs. These genes were located on different chromosomes, most of which on chromosome 12 (22 genes) and the least on chromosomes M (2 genes). GO analysis indicated that the differential methylated genes were involved in axon growth, axon formation, axon elongation and axon guidance. The MAPK, cell adhesion molecules, Ras signaling pathway may be related with the differential methylated genes. Conclusion The methylation levels between ASCs and NSCs are significantly different, which are probably related with axon regeneration.

Objective To study the clinical application of laparoscopic cholecystectomy (LC) combined with intraoperative endoscopic sphincterotomy (IOEST) with the antegrade guidewire technique in the treatment of cholecystocholedocholithiasis.Methods This is a prospective controlled study comparing LC combined with preoperative endoscopic sphincterotomy (POEST) versus LC combined with IOEST in the treatment of cholecystocholedocholithiasis.Patients who were diagnosed to have cholecystocholedocholithiasis from June 2012 to February 2013 in our hospital were divided into the POEST group and the IOEST group.There were 50 patients in each group.The operation time,successful stone-extraction rate,residual stones rate,complication rate,postoperative stay and hospitalization cost between the groups were compared.Results The sex,age,stone size,number of stones and diameter of the common bile duct showed no significant difference between the two groups (P ＞ 0.05).There were no differences between the two groups in surgical time of EST,surgical time of LC combined with EST,successful stone-extraction rate and hospitalization cost (P ＞ 0.05).There were significant differences between the two groups in the residual stones rate,postoperative hyperamylasemia rate,postoperative acute pancreatitis rate and postoperative stay (P ＜0.05).LC combined with IOEST using the antegrade insertion of guidewire technique significantly reduced the residual stones rate (0 vs 8％),postoperative hyperamylasemia rate (4％ vs 18％),postoperative acute pancreatitis rate (0 vs 8％) and postoperative stay.Conclusions LC combined with IOEST using the antegrade guidewire technique could be performed safely when compared with the traditional sequential technique,LC combined with IOEST using the antegrade guidewire technique significantly reduced the postoperative acute pancreatitis rate and the residual stones rate.LC combined with IOEST using the antegrade guidewire technique should be the recommended technique to treat patients with cholecystocholedocholithiasis.

Objective:To explore the expression of mammalian target of rapamycin (mTOR)and its relationgship with the prognosis of breast cancer,and to provide evidence-based basis for the using of mTOR inhibitor in the treatment of breast cancer.Methods: A systemical literature search was conducted based on the following databases:PubMed,EMBbase,Cochrane Library,ISI Web of Science,and CNKI up to November 24,2015.The outcome measures were hazard ratio (HR)with 95% confidence interval (CI) for the association between the mTOR expression and the prognosis of patients with breast cancer.The primary end points including disease-free survival (DFS ), and overall survival (OS ). STATA 12.0 was used to conduct the statistical analysis. Results:A total of seven cohort studies,1 758 patients were included. The risk of recurrence and metastasis of the breast cancer patients with positive expression of mTOR was 2.05 times of the patients with negative expression of mTOR (HR= 2.05, 95% CI: 1.01 - 4.13,P = 0.003);the risk of death in the breast cancer patients with positive expression of mTOR was 2.63 times of the patients with negative expression of mTOR (HR = 2.63, 95%CI:1.45-4.80,P = 0.736).Conclusion:The positive expression of mTOR can significantly increase the recurrence,metastasis and death risk of the patients with breast cancer.

Objective To investigate the clinicopathological features and prognostic factors in patients with low grade glioma. Methods Eighty- seven patients with low grade glioma confirmed by cytological examination were retrospectively analyzed. Results Among 87 patients, male accounted for 62.1%(54/87), and the average age was 36.2 years. The average followed- up time was 51.4 months (3-135 months), and 5 cases were lost in follow-up. The follow- up rate was 94.3% (82/87). The 3- year survival rate was 84.1%, and the 5- year survival rate was 66.7%. The 3- year survival rate was related to clinical symptoms, maximum diameter of tumor and the extent of resection (P<0.01 or<0.05);and the 5- year survival rate was related to years, Karnofsky score, clinical symptoms, maximum diameter of tumor, the pathology classification and extent of resection (P<0.05 or<0.01). Conclusions Low grade glioma is more common in male patients and patients under the age of 40 years. Age below 40 years, Karnofsky score ≥ 70 scores, only epilepsy symptom, tumor diameter below 6.0 cm, oligodendrogliomas, complete resection or subtotal resection are positive prognostic factors. Postoperative radiotherapy can reduce the rate of tumor progression.

Type B aortic dissection is a life-threatening aortic disease.With the development of clinical classification and diagnostic method,the mortality of type B aortic dissection has been greatly decreased.In respect of treatment,endovascular repair due to minimally invasive advantages become the first choice for the treatment of complicated type B aortic dissection.For non-complicated type B aortic dissection,endovascular repair also gradually replace drug treatment,and showed good efficacy.Open surgery is only available for patients who ate not suitable for endovascular repair or repair failure or patients with connective tissue disease.

Objective A total of 102 ESRD patients undergoing hemodialysis for over 3 months in our dialysis center were asked to complete the SF-36 scales.They were also assessed by Hamilton Depression Scale(HAMD17).Univariate analysis was performed to determine the impact of factors such as age,gender,employment status,education,mental status and dialysis status on their quality of life.Methods The 102 patients with ESRD undergoing hemodialysis for over 3 months in our dialysis center completed the SF-36 scales with self-administration,and they were also assessed by Hamilton Depression Scale (HAMD17).Univariate analysis was performed to determine the impact of variables such as age,gender,employment status,education,mental status and dialysis status on the quality of life in patients.Results The scores of PF,RP,BP,GH,VT,SF,RE and MH in patients were significantly lower than those in the normal controls(P

Objective To explore the matrix effect on cyclosporine A (CsA) test by fluorescence polarization immunoassay (FPIA) and enzyme-multiplied immunoassay technique (EMIT), explain the discrepancy of external quality control results between these two methods and find the corrective action.Methods One hundred whole blood samples with various concentrations were adopted and CsA levels were detected by FPIA and EMIT.The results were compared with each other.Moreover, the influence of residual metal ions upon immunoreactions was assessed by adding Cu2+ and Zn2+.The effect of non-whole blood matrix on extraction efficiency for quality control materials and CsA calibrator was evaluated by adding identical volume of Hb-rich reagents followed with re-extraction.Results There is good correlation between results measured with FPIA(X) and EMIT(Y) methods ( Y=0.926 8X -8.115,R2 =0.996 9).Neither FPIA nor EMIT was affected by residual metal ions ( P ＞ 0.05 ). Non-whole blood matrix decreased the extraction efficiency of two methods, but it could be corrected by supplementation of the Hb-rich reagents (≥30 g/L).Conclusions Non-whole blood matrix may be the main reason for the inconsistent results measured by FPIA and EMIT methods.It could be corrected by using Hb-rich reagents.In addition,we should consider the influence of low lib on CsA test,espocially for organ transplant patients with lower Hb ( ＜30 g/L).