2.Clinical study on colonic transmission time and the effect of sini powder on it in functional constipation patients.
Zhao-hui JIN ; Jian-hua DUAN ; Hong-chuan ZHAO
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(10):896-898
OBJECTIVETo investigate the characteristic of colonic transmission in functional constipation (FC) and the effect of traditional Chinese medicine (TCM) Sini Powder (SP) on it.
METHODSThe colonic transmission time (CTT) of 36 patients with FC (the FC group) and 22 healthy subjects (control group) was measured through colonic transmission test, and CTT of entire colon and that of various subsections was calculated with Hinton method and Arhan method respectively. After then, the FC group was treated with SP for 7 days, and CTT was detected again after treatment.
RESULTSBefore treatment, body mass index (BMI) was higher, CTT of entire colon, left half colonic section, and sigmoid-rectal section were longer in the FC group than those in the control group (P < 0.05), no statistical difference in CTT of right half colon was found between the two groups (P > 0.05). After FC patients being treated with SP, their CTT of whole colon, left half colonic section and sigmoid-rectal section were significantly shortened (P < 0.05).
CONCLUSIONFC patients were characterized by increased BMI and CTT prolonged and unevenly distributed in subsections, especially in the left half colon, sigmoid and rectum; SP could shorten the CTT in FC patients.
Adult ; Body Mass Index ; Colon ; drug effects ; physiopathology ; Colon, Sigmoid ; drug effects ; physiopathology ; Constipation ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrointestinal Transit ; drug effects ; Humans ; Male ; Middle Aged ; Phytotherapy ; Time Factors ; Treatment Outcome
4.Identification of Bletillae Rhizoma and its adulterants by SNPs in ITS2.
Dan ZHAO ; Tao ZHOU ; Wei-ke JIANG ; Cheng-hong XIAO ; Chuan-zhi KANG
China Journal of Chinese Materia Medica 2015;40(18):3573-3578
To establish a molecular identification method for Bletillae Rhizoma, this paper extracted genome DNA from Bletillae Rhizoma and its adulterants. The sequences of rDNA ITS2 were sequenced after amplifying. Then multiple alignments of ITS2 were constructed phylogenetic tree with Neighbor Joining by MEGA 5. 1 and found out SNPs loci. The result showed that rDNA ITS2 region could identify Bletillae Rhizoma and its adulterants. There existed the SNPs loci, which could identify Bletilla striata and B. ochracea. Furthermore, we designed specific primers against the SNPs loci of B. striata and B. ochracea, then screened primers and optimized the PCR amplification conditions. Finally, the DNA of B. striata and B. ochracea were specifically amplified by BJ59-412F, BJ59-412R and HHBJ-225R. The length of amplification products were respectively about 350 bp and 520 bp that were effectively identified of B. striata and B. ochracea. While, the adulterants of Bletillae Rhizoma were no-reaction occurring. To sum up, the amplification conditions of the primers can identify B. striata, B. ochracea and their adulterants successfully at the same time. This method was easy, time-saving, and reliable, which can be used as a rapid method for molecular identification of Bletillae Rhizoma.
Base Sequence
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DNA Primers
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genetics
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DNA, Intergenic
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genetics
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DNA, Plant
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genetics
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Drug Contamination
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prevention & control
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Molecular Sequence Data
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Orchidaceae
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classification
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genetics
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Phylogeny
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Polymorphism, Single Nucleotide
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Rhizome
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classification
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genetics
5.Detection of H-ras gene mutations in human breast atypical hyperplasia
Julun YANG ; Ping PUI ; Xuemin CAI ; Li WANG ; Chuan ZHAO ; Lin CAI ; Hong ZOU
Chinese Journal of Clinical and Experimental Pathology 2001;(1):12-14
To investigate the role of H-fas oncogene in the early stages of human breast carcinogenesis. Methods Thirty cases of human breast cancer, 36 epithelial hyperplasia of usual type and 31 atypical hyperplasia were employed to detect H-ras gene codon 12 mutations by PCR-RFLP and PCR-SSCP assays, and to detect the expression of H-ras protein by immunohistochemistry method. ResultsExpression of H-ras protein were found in 73.3 % (22/30) of breast cancer and 48.4 % (15/31 ) of atypical hyperplasia. No H-ras protein expression was observed in hyperplasia of usual type. All tested sarnples of breast cancer and hyperplsia showed no mutations of H-ras gene codon 12. ConclusionOverexpression of H-ras protein is involved in early stages of breast carcinogenesis, but mutations of H-ras gene codon 12 is rarely present in the stage.
6.The effects of rehabilitation training on cardiac function in cerebral infarct patients
Guo-Liang YANG ; Fu-Zhong SI ; De-Yang LI ; Hong GUO ; Jun ZHAO ; Chuan-Xin TIAN ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(10):-
Objective To study the effects of rehabilitation(RT)on cardiac function in cerebral infarct (CIF)patients with cardiac insufficiency(CIS).Methods Fifty-nine CIF patients with CIS were randomly divid- ed into a treatment group(T group,n=29)and a control group(n=30),and all patients were treated with routine pharmacotherapy for 2 months.In addition,RT was administrated in the T group at the same time.The grading of the New York Heart Association(NYHA)and the changes in cardiac function associated index were observed in both groups before and after treatment.Results Compared with the control group,NYHA grades,left ventricle ejection fraction(LVEF),the levels of brain natriuretic peptide(BNP)in the blood plasma,and the 6min walking range of the T group patients were all significantly improved after treatment(P<0.05).Conclusion RT can improve car- diac function in CIF patients with CIS.
7.Cloning of human uracil N-glycosylase and its detection in cancer tissues by quantitative RT-PCR.
Hong-Bo BAO ; Chuan-Bao ZHANG ; Jin-Fang WANG ; Chuan-Nong ZHOU ; Fang LIU ; Xiao-Hang ZHAO ; Shi-Jun QIAN
Chinese Journal of Biotechnology 2003;19(5):561-565
The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell
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genetics
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metabolism
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Cell Line, Tumor
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Esophageal Neoplasms
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genetics
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metabolism
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Humans
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In Vitro Techniques
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Reverse Transcriptase Polymerase Chain Reaction
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Uracil-DNA Glycosidase
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genetics
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metabolism
8.Inhibitory effect of phenylhexyl isothiocyanate on notch signaling of multiple myeloma cells in vitro.
Xiu-Li HONG ; Ze-Chuan ZHANG ; Jiang-Ning ZHAO ; Quan-Yi LU
Journal of Experimental Hematology 2011;19(4):940-943
In order to investigate the mechanisms of phenylhexyl isothiocyanate (PHI) inhibiting the proliferation of multiple myeloma cell RPMI8226 in vitro, the RPMI8226 cells were co-cultured with PHI of various concentrations. The inhibition of proliferation was measured by MTT test and the cell apoptosis was assayed by DAPI staining. The changes of Notch1, Jagged2, BCL-2 and p-Akt proteins in the PHI-treated cells were detected by Western blot. The results showed that PHI inhibited RPMI8226 cell proliferation in certain concentration range and induced their apoptosis. The inhibiting effect caused by PHI showed a concentration-and time-dependent manner. The PHI decreased expressions of Notch1 and Jagged2 proteins in a concentration-and time-dependent manners, the levels of BCL-2 and p-Akt declined at the same time. It is concluded that PHI can inhibit proliferation of RPMI8226 cells, and induce their apoptosis. The cell apoptosis is associated with the inhibition of Notch signaling and downstream targets BCL-2 and p-Akt proteins of RPMI8226 cells, PHI may be a new Notch signaling inhibitor and a promising therapeutic drug for multiple myeloma.
Cell Line, Tumor
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Humans
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Intercellular Signaling Peptides and Proteins
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metabolism
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Isothiocyanates
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pharmacology
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Jagged-2 Protein
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Membrane Proteins
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metabolism
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Multiple Myeloma
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metabolism
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Proto-Oncogene Proteins c-akt
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metabolism
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Proto-Oncogene Proteins c-bcl-2
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metabolism
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Receptor, Notch1
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metabolism
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Signal Transduction
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drug effects
9.Effect of bear bile powder on STAT3 pathway in hepatocellular carcinoma xenograft.
Jin-Yan ZHAO ; Li-Ya LIU ; A-Ling SHEN ; Wei LIN ; Zhi-Yun CAO ; Qun-Chuan ZHUANG ; Zhen-Feng HONG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(8):976-981
OBJECTIVETo observe the effect of bear bile powder (BBP) on the STAT3 pathway and its downstream target genes of nude mice hepatocellular carcinoma (HCC) xenograft, and to explore its mechanism for treating HCC.
METHODSThe subcutaneous xenograft model was established using HepG2 cells. When the subcutaneous transplanted tumor was formed, naked mice were randomly divided into two groups, the BBP group and the control group. Mice in the BBP group were administered with BBP by gastrogavage, once daily for 3 consecutive weeks, while mice in the control group were administered with normal saline by gastrogavage, once daily for 3 consecutive weeks. The body weight and the tumor volume were measured once per week. By the end of medication, the tumor weight was weighed and the tumor inhibition ratio calculated. The apoptosis of the tumor tissue was detected by TdT-mediated dUTP nick end labeling (TUNEL). The expression of Bcl2-associated X protein (Bax), B cell lymphoma/eukemina-2 (Bcl-2), cyclin-dependent protein kinase (CDK4), cyclinD1 were detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of signal transducers and transcription activators 3 (p-STAT3), proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, CDK4, and cyclinD1 were determined by immunohistochemistry.
RESULTSBBP could inhibit the tumor volume and tumor weight, showing statistical difference when compared with the control group (P < 0.01). Results of TUNEL showed that BBP could significantly induce the apoptosis of hepatoma carcinoma cells. Results of RT-PCR showed that BBP could up-regulate the expression of Bax and down-regulate mRNA expression of Bcl-2, CDK4, and cyclinD1. Immunohistochemical results showed that BBP could up-regulate the expression of Bax and inhibit the protein expression of p-STAT3, PCNA, Bcl-2, CDK4, and cyclinD1.
CONCLUSIONBBP could induce the apoptosis of hepatoma carcinoma cells and inhibit their proliferation by regulating STAT3 pathway.
Animals ; Bile ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Ursidae ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism
10.A High-throughput and Quantitative Assay Based on Fluorescence Intensity for Detection of Apoptosis
Ling-Ling YE ; Hong LIU ; Xing-Mao LIU ; Shi-Chong LI ; Ben-Chuan WU ; Qi-Wei WANG ; Zhao-Lie CHEN ;
China Biotechnology 2006;0(06):-
Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P