Simultaneous determination of four kinds of components in Gegen-Huangqin-Huanglian Decoction by UPLC-MS/MS / 国际中医中药杂志

Objective:To develop an UPLC-MS/MS method for the simultaneous determination of baicalin, berberine, glycyrrhizic acid and puerarin in Gegen-Huangqin-Huanglian Decoction. Methods:Isocratic elution was carried out with mobile phase consisting of acetonitrile - 4 mmol/L ammonium formate. The separation was performed on ACE Excel 3 C18 (2.1 mm × 100 mm, 3 μm), and the mass spectrometer was operated in the positive and negative ionization electrospray (ESI) mode using multiple monitoring (MRM) for analysis of four components. External standard method was used for fix quantity. The precursor to product ion transitions monitored for baicalin, berberine, glycyrrhizic acid and puerarin were m/z 447.0→271.0, 336.2→320.2, 821.4→350.9 and 415.2→294.9, respectively. Results:Baicalin, berberine, glycyrrhizic acid and puerarin were all analyzed exactly, the linear ranges were 0.002-0.080, 0.002-0.080, 0.001-0.040, 0.002-0.080 ng, respectively. The r were 0.998 3, 0.999 4, 0.997 9 and 0.999 5, respectively. The recoveries of four analytes ranged from 98.75% to 100.86% and the relative standard deviations were all below 0.74%. Conclusions:UPLC-MS/MS method is sensitive, accuate with rapid speed, which could be used for the determination of baicalin, berberine, glycyrrhizic acid and puerarin in Gegen-Huangqin-Huanglian Decoction.

Simultaneous determination of five compounds in Ganmaoning granules by UPLC-MS/MS / 国际中医中药杂志

Objective To establish a method for the simultaneous determination of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoning granules. Methods The UPLC-MS/MS method was adopted. The separation was performed on ZOBAX SB C18 (2.1 mm×150 mm, 5μm) column with mobile phase consisted of methanol-2 mmol/L ammonium acetate aqueous solution (isocratic elution) at the flow rate of 0.2 ml/min. The column temperature was set at 35℃, and the injection volume was 2μl. The electrospray ionization source (ESI) was used. Multiple reaction monitoring (MRM) mode was adopted, using positive and negative ions scanning. The ion source temperature was 300℃, the drying gas temperature was 400℃, the drying gas flow was 20 L/min, the atomizing gas pressure was 55 psi, and the capillary voltage was 4000 V.Results The linear ranges of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoling granules were 0.20-12.80 ng (r=0.9996), 1.00-64.00 ng (r=0.9998), 0.40-25.60 ng (r=0.9996), 4.00-256.00 ng (r=0.9992), 0.20-12.8 ng (r=0.9991), which showed the linear relationship was good. The limits of detection were 0.02, 0.10, 0.04, 0.40, 0.02 ng, respectively. The limits of quantitation were 0.04, 0.20, 0.08, 0.80, 0.04 ng, separately. TheRSDs of precision, reproducibility and stability (24 h) tests were no more than 3.5％. The recovery rates were 99.3％-100.75％ (RSD=2.09％-3.17％).Conclusions The method established in this experiment is simple, rapid, sensitive and accurate. It can be used to simultaneously determine the contents of five components inGanmaoling granules, and can be used for quality control ofGanmaoninggranules.

Effect of different proportions of piper chinaroot and rhubarb on the dissolution rate of five effective components in rhubarb / 国际中医中药杂志

Objective@#To study the effect of different proportions of piper chinaroot and rheum palmatum on the dissolution rate of five effective components (aloe emodin, emodin acid, emodin, emodin, emodin methyl ether).@*Methods@#The high performance liquid chromatography (HPLC) method was used to analyz the contents of five effective components of rheum palmatum in the extracts of different combination of piper chinaroot and rheum palmatum. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by gradient elution with methanol and 0.1% phosphoric acid water solution as mobile phase at a flow rate of 1ml/min, the column temperature was 30 ℃ and the detection wavelength was 254 nm.@*Results@#The linear ranges of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 0.018 5-0.741 8, 0.017 9-0.717 8, 0.015 9-0.635 5, 0.054 2-2.167 2, 0.016 2-0.646 4 μg, respectively. The average recoveries of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 94.35%, 95.50%, 100.61%, 96.27%, 97.39%, and the RSDs were 1.81%, 1.99%, 2.84%, 2.71%, and 1.86%, respectively. Compared with rheum palmatumby single extract, the content of aloe emodin and emodin were increased by 5 proportions (3:1, 2:1, 1:1, 1:2, 1:3), while the content of emodin and emodin methyl ether were decreased.@*Conclusions@#The optimal compatibility proportion of piper chinaroot and rheum palmatum is 1:1.

Diagnosis and treatment of iatrogenic colonoscopic perforation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To summarize the diagnosis and treatment of iatrogenic colonoscopic perforation (ICP).</p><p><b>METHODS</b>Clinical data, treatment course and outcome of 17 patients who developed ICP following colonoscopic examination or operation at Department of Colorectal Surgery, Changhai Hospital from January 2000 to December 2013 were retrospectively analyzed.</p><p><b>RESULTS</b>During above 13 years, a total of 127 106 patients underwent colonoscopic examination or operation, of whom 17 cases (0.013%) had ICP. There were 8 males and 9 females with an average age of 65.2 (32-85) years. The interval between the onset of ICP and clinically diagnosed ICP was 0 to 6 days after performance. ICP occurred in 8 patients following colonoscopy operations, including simple colonic polyp excision, endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD), while in 9 patients following simple colonoscopy examination. Except for one patient who was immediately diagnosed with ICP through the finding of "yellow adipose tissue visible in the vision field" during operation,7 early cases (41.2%) were diagnosed by abdominal X-ray examination, and 9 later cases were confirmed by abdominal CT examination. The perforation sites included sigmoid colon in 5 cases, caecum in 3 cases, descending colon in 3 cases, descending and sigmoid junction in 2 cases, ileum in 1 case, splenic flexure in 1 case, sigmoid and rectum junction in 1 case, retum in 1 case. One case with ICP following ESD after resection of polyp in caecum was cured successfully with conservative treatment, including fasting, gastrointestinal decompression, fluid infusion, anti-infection and nutritional support. One case with ICP, which was found during colonoscopic operation and the perforation was immediately closed with titanium clip, received conservative treatment, including anti-infection and then was cured. Fifteen patients underwent surgery, including 8 patients with primary intention intestinal perforation repair, 4 patients with primary intention resection of associated intestine and anastomosis, 2 patients with primary intention resection of associated intestine and ostomy, 1 patient with primary intention intestinal perforation repair and ostomy. Postoperative abdominal incision infection occurred in 4 cases, pulmonary infection in 1 case, incision infection with cardiovascular event or urinary tract infection in 1 case each. All the patients were cured and discharged. Average hospital stay was 18.6(3-45) days.</p><p><b>CONCLUSIONS</b>ICP should be diagnosed by physical examination and imaging examination as soon as possible. For perforation during colonoscopic performance, colonoscopic titanium clip can be used for closure. Perforation repair is still the main procedure for ICP. If necessary, partial intestinal resection and anastomosis or ostomy can be selected.</p>

Assessment of the risk factors relating to lymph node metastasis in rectal cancer after neoadjuvant chemoradiotherapy and the clinical significance / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To identify the risk factors associated with lymph node metastasis in rectal cancer after neoadjuvant chemoradiotherapy (CRT).</p><p><b>METHODS</b>From January 2005 to December 2013, the clinical data of 178 patients with advanced rectal cancer undergoing radical excision after neoadjuvant CRT in our department were reviewed retrospectively. A total of 11 clinicopathologic factors relating to lymph node metastasis were studied using univariate and multivariate Logistic regression analyses.</p><p><b>RESULTS</b>There were 74(41.6%) cases with lymph node metastasis, while 104 cases without lymph node metastasis. Univariate analysis showed that age(P=0.000 2), post-CRT CEA level(P=0.011 2), ypT stage(P=0.000 0), pathologic type(P=0.004 0), and tumor regression grade(TRG)(P=0.033 8) were significantly associated with lymph node metastasis. Multivariate analysis showed that age(OR=2.385, 95% CI:1.372 ~ 4.147, P=0.002 1), post-CRT CEA level(OR=2.310, 95% CI:1.005 ~ 5.307, P=0.048 6) and ypT stage(OR=2.592, 95% CI:1.236 ~ 5.432, P=0.011 7) were independent risk factors. However, 15.8% of the patients who achieved TRG1 had lymph node metastasis and TRG failed to independently correlate with lymph node metastasis in rectal cancer after neoadjuvant CRT.</p><p><b>CONCLUSIONS</b>There was a higher ratio of lymph node metastasis in rectal cancer patients who were young, CEA≥5 μg/L or deep invasion after neoadjuvant CRT. Therefore, neoadjuvant CRT should be carefully considered in these patients.</p>

Expression of BRAF V600E mutant protein in cutaneous malignant melanoma / 中华皮肤科杂志

Objective To detect the expression of BRAF V600E mutant protein in cutaneous malignant melanoma (CMM),and to evaluate the sensitivity and specificity of immunohistochemistry (IHC) in detecting BRAF V600E mutation.Methods IHC with an anti-BRAF V600E monoclonal antibody was performed to detect the expression of BRAF V600E mutant protein in paraffin-embedded tissue sections from 103 patients with CMM and 40 patients with nevus.Statistical analysis was carried out with SPSS software version 17.0,and the expression rate of BRAF V600E mutant protein was compared by chi-square test.Results The expression rate of BRAF V600E mutant protein in the CMM patients was 20.4％ (21/103),significantly higher than that in the nevus patients (5.0％ (2/40),x2 =5.06,P ＜ 0.05).Significant differences were observed in the expression rate of BRAF V600E mutant protein between CMM patients of different age groups (29.8％ (14/47) in patients aged ＜ 60 years vs.12.5％ (7/56) in those aged ≥ 60 years,P ＜ 0.05) and nationality (30.2％ (13/43) for Uygur nationality vs.13.3％ (8/60) for Han nationality,P ＜ 0.05),as well as among CMM lesions from different anatomical sites (13.6％ (6/42) in acral sites vs.11.8％ (4/29) in mucous membrane vs.45.8％ (11/32) in non-acral sites,P ＜ 0.05) and of different Clark levels (8.6％ (4/42) for grade Ⅰ-Ⅲ vs.12.4％ (17/61) for grade Ⅳ-Ⅴ,P＜ 0.05),but not between male and female CMM patients or between CMM patients with lymph node metastasis and those without (both P ＞ 0.05).IHC with the anti-BRAF V600E antibody showed a sensitivity of 100％ (15/15) and a specificity of 98.5％ (65/66) in detecting BRAF V600E mutation.Conclusions The expression of BRAF V600E mutant protein is up-regulated in CMM lesions,and CMM patients of Uygur nationality seems to have a higher expression rate than those of Han nationality.IHC appears to be an accurate and rapid method to detect V600E BRAF mutation.