1.Molecular identification of Cynomorii herba using ITS2 DNA barcoding.
Dian-Yun HOU ; Jing-Yuan SONG ; Lin-Chun SHI ; Pei YANG ; Shi-Lin CHEN ; Hui YAO
China Journal of Chinese Materia Medica 2013;38(23):4028-4032
OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.
METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.
RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.
CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.
Cynomorium ; classification ; genetics ; DNA Barcoding, Taxonomic ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Polymerase Chain Reaction
2.Observation on effect of danxiaoling in supplementary treatment of intrahepatic cholestasis in pregnancy.
Dian-yun SHI ; Hua CHEN ; Mei XIAO
Chinese Journal of Integrated Traditional and Western Medicine 2002;22(2):116-118
OBJECTIVETo evaluate the curative effect of Danxiaoling Pill (DXLP), a Chinese herbal preparation, in treating intrahepatic cholestasis in pregnancy (ICP).
METHODSFifty-eight cases of ICP were divided randomly into two groups and treated by DXLP and Composite Yiganling as control respectively with the other identical conventional treatment. The changes of clinical symptoms, related laboratory parameters after treatment and the condition of labor were observed.
RESULTSThe total effective rate in both groups was 100%, but the markedly effective rate in the DXLP treated group was higher than that in control group (P < 0.01). Levels of blood cholyglycine acid (CGA), alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin, total cholesterol and low density lipoprotein were all decreased in both groups after treatment, but DXL showed a better efficacy in decreasing CGA, ALT and AST than that of Yiganling. Moreover, the amniotic fluid meconium contaminated rate, premature delivery occurrence in the DXLP group were lower than those in the control group, while the weight of newborn baby was higher in the former than in the latter.
CONCLUSIONDXLP could effectively lower the serum bile acid and improve liver function in treating ICP.
Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Cholestasis, Intrahepatic ; blood ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glycocholic Acid ; blood ; Humans ; Phytotherapy ; Pregnancy ; Pregnancy Complications ; blood ; drug therapy
3.Clinical and experimental study on wuling pill in treating gestation period intrahepatic cholestasis.
Dian-yun SHI ; Hua CHEN ; Xiao-yun TIAN
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(2):114-118
OBJECTIVETo evaluate the curative effect of Wuling pill (WLP), a traditional Chinese patent medicine, in treating gestation period intrahepatic cholestasis (GPI).
METHODSIn the clinical study, 90 GPI patients were divided into the treated group treated by conventional therapy plus WLP and the control group treated by conventional therapy plus compound Yiganling (YGL) with a ratio of 2:1. Clinical symptoms and accouchement condition were observed. Levels of cholyglycine acid (CGA), total and direct bilirubin (TB and DB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were detected before and after treatment. For the experimental study, GPI rat model was induced by injecting estradiol benzoate to pregnant Wistar rats. The model rats in the treated group were administrated with WLP by gastro-perfusion and those in the control group, were administrated with YGL. Levels of CGA,TB,DB, ALT, AST and ALP in the mother and fetal rats, as well as in the amnionic fluid were determined. Besides, the volume of bile excreted by the mother rats was observed.
RESULTSIn clinical trials, the markedly effective rate in the treated group (47 cases, 78.3% ) was higher than that in the control group (15 cases, 50%, chi2 = 7.17286, P < 0.01). Levels of blood CGA, TB, ALT and AST were all decreased in both groups after treatment, but WLP showed a better efficacy than YGL (P < 0.05) in lowering CGA, ALT and AST. Moreover, the occurrence of meconium contaminated amnionic fluid and premature delivery were lower, while weight and Apgar grade of newborn babies were higher in the treated group than those in the control group. In animal experiment, WLP showed significant effects in decreasing CGA level in amniotic fluid, and in blood of the mother and fetal rats. In addition, it could also decrease the levels of bilirubin, ALT and AST, and promote the bile excretion to reduce CGA concentration in bile. All the above effects showed a dose-dependent pattern.
CONCLUSIONWLP could effectively lower the serum bile acid, improve the hepatic function and better the pregnant outcome in treating GPI.
Adult ; Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; Cholestasis, Intrahepatic ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glycocholic Acid ; blood ; Humans ; Phytotherapy ; Pregnancy ; Pregnancy Complications ; drug therapy ; Pregnancy Outcome ; Rats ; Rats, Wistar
4.The infection status of anisakid larvae in marine fish and cephalopods from the Bohai Sea, China and their taxonomical consideration.
Hong Wei MA ; Tai Jing JIANG ; Fu Shi QUAN ; Xiao Guang CHEN ; Hui dong WANG ; Yun Shu ZHANG ; Ming Shan CUI ; Wen Yan ZHI ; Dian Chen JIANG
The Korean Journal of Parasitology 1997;35(1):19-24
The infection status of anisakid larvae was examined in 290 marine fish of 25 species and in 108 cephalopods of 3 species purchased in Bayuquan region, Yingko city nearby the coast of the Bohai Sea from may to August 1992. A total of 7,327 larvae were collected from 156 fish of 19 species and 8 squids of one species. The 3rd-stage larvae of Anisakis simplex were collected from 121 fish (63.4%) of 15 species (N = 191) and from 8 squids (14.8%) of one species (N = 54), and they were total, 5,992 (81.8%). Out of remaining 1,335 larvae, 154 (2.1%) were classified as Thynnascaris type B from 23 fish of 4 species, 1,013 (13.8%) as Thynnascaris type C from 79 fish of 13 species. 164 (2.2%) as Hysterothylacium China type V from 20 fish of 4 species, 3 (0.04%) as Raphidascaris from 3 fish of 2 species and one was Pseudoterranova decipiens larva.
Animal
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Anisakiasis/veterinary*
;
Anisakiasis/parasitology
;
Anisakiasis/epidemiology
;
Anisakis/isolation & purification
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Anisakis/classification*
;
China
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Fish Diseases/parasitology*
;
Fish Diseases/epidemiology
;
Fishes
;
Larva
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Seawater
;
Squid/parasitology*
5.Microvascular decompression for trigeminal neuralgia in elderly patients
Sheng-Wei QIAN ; Yun-Ping DUAN ; Ji-Zhi ZHANG ; Dian-Shi JIN
Chinese Journal of Neuromedicine 2009;8(4):399-401,404
Objective To assess the efficacy and safety of microvascular decompression for treatment of trigeminal neuralgia in elderly patients. Methods The clinical data of29 elderly patients (above 65 years old) with trigeminal neuralgia treated with microvascular decompression were retrospectively reviewed to analyze the outcomes of the surgery and complications in comparison with those of younger patients below 65 years of age. Results Good therapeutic effects were achieved in both groups without obvious difference between them,the rate ofpostoperative pain relieffor the elderly group is 93.10%,96.36%for the control group.The elderly patients showed some special anatomical and systemic features in relation to aging,including the presence of brain atrophy,obvious arteriosclerosis in the culprit arteries(the elderlygroupis about 1/4,the control group is about 1/10),multiple systemic diseases(the elderly group accounted for 71.69%,41.81%for control group),and increased risk of surgical complications(one of the elderly group appears brainstem infarction,2 cases of lower extremity deep VenOUS thromboembolism disease). Conclusion Microvascular decompression provides an effective cure or pain relief for most elderly patients with trigeminal neuralgia with the application of effective perioperative treatment.
6.Molecular identification of Corni Fructus and its adulterants by ITS/ITS2 sequences.
Dian-Yun HOU ; Jing-Yuan SONG ; Hui YAO ; Jian-Ping HAN ; Xiao-Hui PANG ; Lin-Chun SHI ; Xiao-Chen WANG ; Shi-Lin CHEN
Chinese Journal of Natural Medicines (English Ed.) 2013;11(2):121-127
UNLABELLED:
The DNA barcoding method was used to accurately and rapidly identify Corni Fructus and its adulterants.
METHODS:
Genomic DNA extracted from Corni Fructus and its adulterants were used as templates. The ITS (internal trascribed spacer) regions were amplified using polymerase chain reaction. Sequence assembly was performed using CodonCode Aligner V 3.5.4. Genetic distances were computed using MEGA V 5.0. Species identification was conducted using neighbor-joining (NJ) trees.
RESULTS:
The ITS sequence length of Corni Fructus was 659 bp. The average intra-specific genetic distance of Corni Fructus was 0.005, markedly lower than the inter-specific genetic distance between Corni Fructus and its adulterants (0.357). The ITS2 sequence length of Corni Fructus was 250 bp. No variation was found among the different samples. The interspecific genetic distance of ITS2 between Corni Fructus and its adulterants was 0.571. NJ trees and BLAST results indicated that Corni Fructus and its adulterants can be easily differentiated with monophyly.
CONCLUSION
ITS/ITS2 regions can accurately and efficiently distinguish Corni Fructus and its adulterants. In addition, the results not only established the foundation for the clinical safety in the utilization of Corni Fructus, but also provided reference for molecular identification of other Chinese herbal medicine and Chinese herbal pieces.
Base Sequence
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Cornus
;
classification
;
genetics
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DNA, Plant
;
genetics
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DNA, Ribosomal Spacer
;
genetics
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Drug Contamination
;
Molecular Sequence Data
;
Molecular Typing
;
methods
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Phylogeny
;
Species Specificity
7.Targeted degradation of androgen receptors in androgen-independent prostate cancer cells: an experimental study.
Yun-Feng SHI ; Yue-Qing TANG ; Xiao-Wen SUN ; Dian-Jun YU ; Bang-Min HAN ; Yan HONG ; Shu-Jie XIA
National Journal of Andrology 2009;15(12):1059-1063
OBJECTIVETo investigate targeted degradation of the androgen receptor (AR) by chimeric molecules (DHT-PROTAC) via the ubiquitin-proteasome pathway in androgen-independent prostate cancer CA-2B cells, and explore the proliferation, secretion and apoptosis of the treated cells.
METHODSC4-2B cells were treated with DHT-PROTAC, and then the expressions of the AR protein and caspase3 in the C4-2B cells were detected by immunohistochemistry and Western blot. The concentration of PSA in the supernatant was examined by ELISA. The cells were counted and their proliferation analyzed by a growth curve. The inhibitory effect on the growth of C4-2B cells was evaluated by MIT assay.
RESULTSCompared with the control group, the DHT-PROTAC-treated group showed an obviously decreased expression of AR proteins with a significant attenuation of the band signals (P < 0.05), a 40% reduction of the AR-positive cells and a 60% decrease of the PSA concentration in the supernatant (P < 0.05). DHT-PROTAC exhibited an inhibitory effect on the C4-2B cells in a time-dependant manner (P < 0.05).
CONCLUSIONThe chimeric molecule (DHT-PROTAC) can target the degradation of androgen receptors, reduce the secretion of PSA and repress the in vitro growth of C4-2B cells.
Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Male ; Prostate-Specific Antigen ; metabolism ; Prostatic Neoplasms ; drug therapy ; metabolism ; pathology ; Receptors, Androgen ; metabolism
8.Proportion of intermediate epithelial cells and human prostate cancer.
Dian-Jun YU ; Yue-Qing TANG ; Yun-Feng SHI ; Yong-Chuan WANG ; Jian ZHUO ; Yi-Yong ZHU ; Xiao-Wen SUN ; Shu-Jie XIA
National Journal of Andrology 2010;16(12):1063-1067
OBJECTIVETo study the different proportions of intermediate epithelial cells in human prostate cancer tissue and their clinical significance.
METHODSWe performed immunohistochemical staining for Cytokeratin 5 (CK5) and Cytokeratin 8 (CK8) on 60 samples of human prostate cancer, determined the proportions of intermediate epithelial cells in the cancer tissue, and classified the samples into 2 types, one with a majority of intermediate epithelial cells (CaP-INT, n = 32), and the other composed mostly of luminal epithelial cells (CaP-LUM, n = 28). Then we compared the 2 types of prostate cancer in the expression of the androgen receptor (AR), age of the patient, serum t-PSA, prostate volume, Gleason score, clinical stage, androgen resistance, and incidence of distant metastasis.
RESULTSCaP-INT showed a significantly lower expression of AR ([24.42 +/- 11.41] %) and a higher incidence of distant metastasis (n = 14) than CaP-LUM ([77.21 +/- 10.22] % and n = 4) (P < 0.05). In the CaP-INT group, 6 of the 26 endocrinologically treated cases developed into androgen-independent prostate cancer (AIPC), while in the CaP-LUM group, only 1 out of 23 (P < 0.05). The former also showed remarkably higher clinical stages than the latter (P < 0.05), but no significant differences were found in age, serum t-PSA, prostate volume and Gleason score between the two groups (P > 0.05).
CONCLUSIONA higher proportion of intermediate epithelial cells may lead to increased invasiveness and metastasis of human prostate cancer.
Aged ; Aged, 80 and over ; Cell Count ; Cell Differentiation ; Epithelial Cells ; classification ; pathology ; Humans ; Male ; Middle Aged ; Prostate ; pathology ; Prostatic Neoplasms ; pathology ; Receptors, Androgen ; metabolism
9.Principles for molecular identification of traditional Chinese materia medica using DNA barcoding.
Shi-Lin CHEN ; Hui YAO ; Jian-Ping HAN ; Tian-Yi XIN ; Xiao-Hui PANG ; Lin-Chun SHI ; Kun LUO ; Jing-Yuan SONG ; Dian-Yun HOU ; Shang-Mei SHI ; Zhong-Zhi QIAN
China Journal of Chinese Materia Medica 2013;38(2):141-148
Since the research of molecular identification of Chinese Materia Medica (CMM) using DNA barcode is rapidly developing and popularizing, the principle of this method is approved to be listed in the Supplement of the Pharmacopoeia of the People's Republic of China. Based on the study on comprehensive samples, the DNA barcoding systems have been established to identify CMM, i.e. ITS2 as a core barcode and psbA-trnH as a complementary locus for identification of planta medica, and COI as a core barcode and ITS2 as a complementary locus for identification of animal medica. This article introduced the principle of molecular identification of CMM using DNA barcoding and its drafting instructions. Furthermore, its application perspective was discussed.
Animals
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China
;
DNA
;
genetics
;
DNA Barcoding, Taxonomic
;
methods
;
DNA, Ribosomal Spacer
;
genetics
;
Drugs, Chinese Herbal
;
classification
;
isolation & purification
;
Electron Transport Complex IV
;
genetics
;
Materia Medica
;
classification
;
isolation & purification
;
Medicine, Chinese Traditional
;
Plant Proteins
;
genetics
;
Plants, Medicinal
10.Surgical treatment of giant coronary artery aneurysm.
Dian-yuan LI ; Shang-shou HU ; Li-zhong SUN ; Yun-hu SONG ; Wei WANG ; Shi-wei PAN ; Hong ZHAO ; Guo-hua LUO ; Yong-min LIU ; Zhi-tao QI ; Tian-fu TAO
Chinese Journal of Surgery 2006;44(22):1535-1537
OBJECTIVETo discuss the clinical features and surgical treatments of giant coronary artery aneurysm (CAA).
METHODSFrom July 1996 to October 2004, 6 giant CAA patients were underwent surgery at Fuwai hospital. Three cases were underwent CAA resection, 2 concomitant coronary bypass, 3 reconstruction. The giant CAA was often combined with other cardiac diseases. Four cases underwent additional procedures of fistula closure, 3 aortic valve replacements, 2 aortoplasty and 1 thrombus cleaning at the same time.
RESULTSAll patients recovered uneventfully. The mean of cardiopulmonary bypass time was (144 +/- 26) min (range 67 to 207 min). Aortic cross clamping time was (104 +/- 21) min (range 56 to 172 min). Patients follow-up time occurred from 8 to 87 months (mean of 48 months). All patients were free of symptoms during follow-up. None of the patients died during the follow-up period and none of the CAA recurred.
CONCLUSIONSThe giant CAA is a serious cardiovascular disease, early diagnosis and surgical treatment are mandatory.
Adult ; Coronary Aneurysm ; pathology ; surgery ; Coronary Artery Bypass ; Coronary Vessels ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Surgical Procedures, Operative ; methods ; Treatment Outcome