2.Comparison of real-time PCR method with Sanger sequencing for detection of BRAF muta-tion in papillary thyroid carcinoma
Tian QIU ; Wenting HUANG ; Lei GUO ; Haizhen LU ; Yun LING ; Ling SHAN ; Wenbin LI ; Ning Lü ; Jianming YING
Chinese Journal of Clinical and Experimental Pathology 2015;(7):756-758
Purpose To investigate the positive rate and concordance rate of BRAF mutation in papillary thyroid carcinoma detected by real-time PCR method and Sanger sequencing. Methods 312 papillary thyroid carcinomas patients were enrolled in this study. Real-time PCR method and Sanger sequencing were performed to detect BRAF gene mutations. The frequency of BRAF mutation and the concordance of two methods were analyzed. Results BRAF mutation was detected in 65. 4% (204/312) and 63. 8% (199/312) of 312 papillary thyroid carcinoma samples by using real-time PCR method and Sanger sequencing, respectively. There was no significant correlation between BRAF gene mutations and patients’ gender. There was significant correlation between BRAF gene mutations and patients’ age. The overall concordance between real-time PCR method and Sanger sequencing for BRAF mutation detection was 98. 4%. Conclusion Real-time PCR method provides an effective method in BRAF gene mutation detection.
3.Chemical constituents from roots of Illicium majus.
Chang-Shan NIU ; Ya-Dan WANG ; Jing QU ; Shi-Shan YU ; Yong LI ; Yun-Bao LIU ; Shuang-Gang MA ; Hai-Ning LV ; Xia CHEN ; Song XU
China Journal of Chinese Materia Medica 2014;39(14):2689-2692
Ten compounds, including seven sesquiterpenes, two phenols and one phenylpropanoid, were isolated from the roots of Illicium majus by means of silica gel, ODS, Sephadex LH-20, and preparative HPLC. On analysis of MS and NMR spectroscopic data , their structures were established as cycloparviflorolide (1), cycloparvifloralone (2), tashironin (3), tashironin A (4), anislactone A(5), anislactone B (6), pseudomajucin (7), syringaldehyde (8), methyl-4-hydroxy-3, 5-dimethoxybenzoate (9), and (E)-3-methoxy-4,5-methylenedioxycinnamic alchol (10), respectively. Compounds 1-4 and 8-10 were first isolated from this plant. In the in vitro assays, at a concentration of 1.0 x 10(-5) mol x L(-1), compounds 5 and 6 were active against LPS induced NO production in microglia with a inhibition rate of 75.31% and 53.7%, respectively.
Drugs, Chinese Herbal
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analysis
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chemistry
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Illicium
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chemistry
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Organic Chemicals
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analysis
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chemistry
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Plant Roots
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chemistry
4.Childhood-onset myasthenia gravis: the analysis of influencing factors of therapeutic effect and prognosis.
Ning-ning QIN ; Rui-ling CHEN ; Shan MA ; Xiao-jun ZHANG ; Hou-liang SUN ; Yun JING
Chinese Journal of Pediatrics 2011;49(5):371-375
OBJECTIVEThough myasthenia gravis (MG) is a typical autoimmune disorder, there was some controversy on the treatment of the childhood-onset MG. By observing the efficacy of different therapies, the authors analyzed the affecting factors of prognosis in childhood-onset MG.
METHODThe retrospective data of 155 patients with childhood-onset MG (age of MG onset was less than 15 years) were collected from Department of Neurology, Beijing Tongren Hospital (January 2000 - February 2010). The patients were non-randomly divided according to their treatment into 3 groups (glucocorticoid, thymectomy and glucocorticoid combined with thymectomy groups). Postintervention status meeting the criteria of Myasthenia Gravis Foundation of America (MGFA) "complete stable remission, CSR", "pharmacologic remission, PR", "minimal manifestations, MM", or "Improved, I" was regarded as desirable response, which was used as primary indicator of observation. The authors assessed the efficacy of three therapies and analyzed the influencing factors of prognosis by using Chi-square test and Logistic regression.
RESULTAt 3 months of treatment, glucocorticoid group showed the highest effective rate. At the end of 1 year or 2 years of treatment, glucocorticoid combined with thymectomy group showed the highest effective rate respectively. The generalization rate of MG at 2 years, 10 years and 20 years in childhood-onset ocular MG patients were 4.3%, 10.7%, and 41.5%, respectively. Of patients with generalization of MG, 48.1% occurred within 2 years, 92.6% within 20 years. Univariate analysis showed that in childhood-onset ocular MG patients, variables such as age at onset (> 10 years), LG-MG and with chronic fatigue were significantly associated with general MG conversion. Whereas multivariate analysis showed that patients with age at onset (> 10 years) and chronic muscle fatigue were apt to convert to generalized MG.
CONCLUSIONGlucocorticoid appeared to have an effect that leads to early remission of symptoms in childhood-onset MG patients and glucocorticoid combined with thymectomy appeared to have better long-term effect. For those childhood-onset ocular MG patients with longer course of disease, older age of onset, chronic fatigue, or LG-MG, physicians should try to prevent the generalization of MG by immunosuppressive therapies.
Adolescent ; Child ; Child, Preschool ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Myasthenia Gravis ; diagnosis ; therapy ; Prognosis ; Retrospective Studies ; Thymectomy ; Young Adult
5.Clinical pathological analysis of rectal cancer lymph node skip metastasis.
Xiao-hui DU ; Ning NING ; Yun-shan ZHAO
Chinese Journal of Surgery 2012;50(3):219-221
OBJECTIVETo analyze clinical and pathological of lymph node skip metastasis of rectal cancer and discuss the meaning of the high vascular ligation.
METHODSA retrospective analysis of 207 cases for radical resection of rectal cancer was made, meanwhile the skip metastasis of the roots of the inferior mesenteric artery lymph nodes was studied. Combined with clinical data, the relevance of clinical and pathological factors with the skip metastasis was analyzed.
RESULTSThe 207 cases of rectal cancer patients surgical resection specimen detected 2305 pieces of lymph node, the transfer of 168 patients with. The statistical analysis found that skip metastasis related with tumor differentiation (χ(2) = 113.65, P = 0.037) and depth of tumor invasion (χ(2) = 108.22, P = 0.042), but gender, age, location, size, preoperative carcinoembryonic antigen level, gross type and tissue types factors were not significantly correlation.
CONCLUSIONSPreoperative differentiation of cancer and tumor invasion depth assessment can help prompt the existence of lymph node skip metastasis. The assessment of the risk of skip metastasis for patients should be performed the high vascular ligation and lymph node dissection.
Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Rectal Neoplasms ; pathology ; surgery ; Retrospective Studies
6.Applications and prospect of multiple displacement amplification in preimplantation genetic diagnosis.
Yin-feng ZHANG ; Hai-ning LUO ; Xiao-pei LI ; Yun-shan ZHANG
Chinese Journal of Medical Genetics 2012;29(4):431-434
Multiple displacement amplification (MDA) is a new technology for whole genome amplification (WGA), which can generate large amount of high-quality DNA and features high amplification efficiency and fidelity. MDA combined with conventional PCR techniques has been successfully applied for preimplantation genetic diagnosis, which has broaden latter's clinical applications.
Genome, Human
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Humans
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Nucleic Acid Amplification Techniques
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methods
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Polymerase Chain Reaction
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methods
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Preimplantation Diagnosis
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methods
7.Comparison of real-time PCR-optimized oligonucleotide probe method and Sanger sequencing for detection of KRAS mutations in colorectal and lung carcinomas.
Tian QIU ; Yun LING ; Zhao CHEN ; Ling SHAN ; Lei GUO ; Ning LÜ ; Jian-ming YING
Chinese Journal of Pathology 2012;41(9):599-602
OBJECTIVETo investigate the feasibility of real-time PCR-optimized oligonucleotide probe method for detection of KRAS mutations in lung and colorectal carcinomas, as compared with Sanger sequencing method.
METHODSGenomic DNA was extracted from formalin fixed paraffin embedded samples of 221 lung carcinomas and 131 colorectal carcinomas after tumor cell content assessment and macrodissection. Real-time PCR-optimized oligonucleotide probe method and Sanger sequencing were performed to detect KRAS gene mutations. The frequency of KRAS mutation, mutation types, and their concordance were analyzed.
RESULTSKRAS mutation was detected in 6.3% (14/221) and 4.5% (10/221) of 221 lung cancer samples by using real-time PCR-optimized oligonucleotide probe method and Sanger sequencing, respectively, while in 41.2% (54/131) and 40.5% (53/131) of 131 colorectal cancer samples, respectively. There was no significant correlation between KRAS gene mutations and patients' gender and age (P > 0.05). The positive rate of KRAS codon 12 mutation was significantly higher than that of KRAS codon 13 mutation (P < 0.05). The overall concordance between real-time PCR-optimized oligonucleotide probe method and Sanger sequencing for KRAS mutation detection was 97.4%.
CONCLUSIONReal-time PCR-optimized oligonucleotide probe method provides an alternative method with high consistency and sensitivity as compared to Sanger sequencing in gene mutation detection.
Codon ; Colorectal Neoplasms ; genetics ; DNA Mutational Analysis ; methods ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Oligonucleotide Probes ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins p21(ras) ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; ras Proteins ; genetics
8.Detection of KRAS, BRAF, PIK3CA and EGFR gene mutations in colorectal carcinoma.
Yun LING ; Jian-ming YING ; Tian QIU ; Ling SHAN ; Lei GUO ; Ning LÜ
Chinese Journal of Pathology 2012;41(9):590-594
OBJECTIVETo investigate the mutation frequencies of KRAS, BRAF, PIK3CA and EGFR genes that were effective on the targeted therapies in colorectal carcinoma.
METHODSThe tissue specimens from 331 colorectal cancer patients were collected and subject to KRAS, BRAF, PIK3CA and EGFR mutation analysis. Paraffin-embedded tissue samples were obtained and macrodissection was performed to enrich the tumor cells for DNA extraction when necessary. PCR-based direct DNA sequencing was used to investigate the codons 12 and 13 in exon 2 of KRAS gene, exons 11 and 15 of BRAF gene, exons 9 and 20 of PIK3CA gene and exons 18-21 of EGFR gene.
RESULTSActivating mutations were detected in KRAS (44.1%, 137/311), BRAF (5.8%, 9/156), PIK3CA (2.6%, 4/156) and EGFR (1.3%, 2/156) in the study cohort of colorectal carcinoma cases. Among KRAS gene mutations, 81.0% (111/137) occurred in codon 12, with p.G12D as the most common variant (45.3%, 62/137); 19.0% (26/137) occurred in codon 13, with 38G > A (G13D) as the most common variant (17.5%, 24/137).
CONCLUSIONSThe KRAS mutation frequency is the highest among the four genes (KRAS, BRAF, PIK3CA and EGFT) tested in colorectal carcinoma. The presence of these gene mutations may provide therapeutic information for targeted therapy. Mutation analyses of BRAF and PIK3CA in addition to KRAS should be a part of the standard diagnostic algorithm for colorectal carcinoma patients.
Class I Phosphatidylinositol 3-Kinases ; Codon ; Colorectal Neoplasms ; genetics ; pathology ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Phosphatidylinositol 3-Kinases ; genetics ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins B-raf ; genetics ; Proto-Oncogene Proteins p21(ras) ; Receptor, Epidermal Growth Factor ; genetics ; ras Proteins ; genetics
9.Robotic anterior resection of rectal cancer: technique and early outcome.
Xiao-hui DU ; Di SHEN ; Rong LI ; Song-yan LI ; Ning NING ; Yun-shan ZHAO ; Zhen-yu ZOU ; Na LIU
Chinese Medical Journal 2013;126(1):51-54
BACKGROUNDThe Da Vinci system is a newly developed device for colorectal surgery. With advanced stereoscopic vision, lack of tremor, and the ability to rotate the instruments surgeons find that robotic systems are ideal laparoscopic tools. Since conventional laparoscopic total mesorectal excision is a challenging procedure, we have sought to assess the utility of the Da Vinci robotic system in anterior resections for rectal cancer.
METHODSBetween November 2010 and December 2011, a total of 22 patients affected by rectal cancer were operated on with robotic technique, using the Da Vinci robot. Data regarding the outcome and pathology reports were prospectively collected in a dedicated database.
RESULTSThere were no conversions to open surgery and no postoperative mortality of any patient. Mean operative time was (220 ± 46) minutes (range, 152 - 286 minutes). The median number of lymph nodes harvested was (14.6 ± 6.5) (range, 8 - 32), and the circumferential margin was negative in all cases. The distal margin was (2.6 ± 1.2) cm (range, 1.0 - 5.5 cm). The mean length of hospital stay was (7.8 ± 2.6) days (range, 7.0 - 13.0 days). Macroscopic grading of the specimen was complete in 19 cases and nearly complete in three patients.
CONCLUSIONSRobotic anterior resection for rectal surgery is safe and feasible in experienced hands. Outcome and pathology findings are comparable with those observed in open and laparoscopy procedures. This technique may facilitate minimally invasive radical rectal surgery.
Aged ; Digestive System Surgical Procedures ; instrumentation ; methods ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Robotics ; methods ; Treatment Outcome
10.Qilin pills combined with bromocriptine for idiopathic hyperprolactinemic oligoasthenozoospermia.
Yun-Shan DENG ; Jin-Fang ZHOU ; Xia-Yun LI ; Xun-Ning WU ; Qi-Sheng GAN ; Yao-Fan WU ; Feng-Ying SU
National Journal of Andrology 2013;19(10):940-944
OBJECTIVETo observe the therapeutic effect of Qilin Pills combined with bromocriptine on idiopathic hyperprolactinemic (HPRL) oligoasthenospermia.
METHODSWe conducted a randomized controlled study on 40 cases of idiopathic HPRL oligoasthenospermia, who were equally assigned to a trial group and a control group to be treated with Qilin Pills (6 g tid) combined with bromocriptine and bromocriptine alone, respectively, both for a course of 12 weeks. Then we observed the changes in the semen volume, sperm concentration, sperm motility and the levels of serum prolactin and testosterone, and compared the therapeutic results between the two groups before and after medication.
RESULTSCompared with the parameters before medication, both the trial and the control group showed significant improvement after treatment in sperm concentration ([11.60 +/- 3.90] x 10(6)/ml vs [28.10 +/- 13.50] x 10(6)/ml and [12.03 +/- 4.10] x 10(6)/ml vs [18.85 +/- 8.50] x 10(6)/ml), the percentage of grade a sperm ([8.75 +/- 6.65]% vs [24.35 +/- 13.25 ]% and [8.70 +/- 6.70] % vs [19.65 +/- 10.05]%), the percentage of grade a + b sperm ( [28.45 +/- 11.35]% vs [45.80 +/- 16.55]% and [27.65 +/- 10.65]% vs [35.66 +/-13.25]%), and sperm motility ([38.22 +/- 16.35]% vs [60.05 +/- 20.65]% and [37.25 +/- 15.75 ]% vs [52.65 +/- 18.25 ]%) (all P<0.01). No significant differences were found in semen volume (P>0.05). The serum prolactin levels were significantly decreased in the trial and control groups ([152.00 +/- 22.32] and [160.45 +/- 26.65] mIU/L), as compared with premedication ([482.25 +/- 65.32] and [477.32 +/- 60.25] mIU/L) (P<0.01), while the serum testosterone levels were remarkably higher ([16.35 +/- 5.52] and [11.15 +/- 4.65] nmol/L) than before treatment ([3.75 +/- 1.10] and [4.05 +/- 1.30] nmol/L) (P<0.01). There were no statistically significant differences in the serum prolactin and testosterone levels between the two groups after treatment (P>0.05).
CONCLUSIONQilin Pills combined with bromocriptine have a significantly better efficacy than bromocriptine alone in the treatment of idiopathic HPRL oligoasthenospermia.
Adult ; Asthenozoospermia ; blood ; drug therapy ; Bromocriptine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Hyperprolactinemia ; blood ; drug therapy ; Male ; Oligospermia ; blood ; drug therapy ; Phytotherapy ; Prolactin ; blood ; Young Adult