2.Combination of cytogenetic analysis and molecular screening in patients with de novo acute myeloid leukemia.
Zhe, GENG ; Heng, ZHANG ; Di, WANG ; Yi, XIAO ; Na, WANG ; Chunrui, LI ; Liang, HUANG ; Jianfeng, ZHOU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(4):501-10
Nowadays the role of genetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukemia (AML) has become more valuable. To improve and validate the detection of clonal chromosomal aberrations in leukemia, we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML. Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients, and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH. The PCR products in some suspected cases were tested by two-directional sequencing. The results showed that except unqualified samples, fusion genes were detected by multiplex RT-PCR in 211 of 474 patients (44.51%), including AML1-ETO, CBFβ-MYH11, PML-RARα, PLZF-RARα, NPM-RARα, MLL rearrangements, BCR-ABL, DEK-CAN, SET-CAN, TEL-PDGFR, TLS-ERG, AML1-MDS1 (EVI-1). In 373 patients, who took both multiplex RT-PCR and karyotype analysis, the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373 (42.89%) and 179/373 (47.98%) respectively, and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373 (57.90%). The PCR results from 11 cases "normal" in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing. It is concluded that karyotype studies remain the cornerstone for genetic testing; conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML, especially for the cryptic or submicroscopic aberrations. Once a genetic marker has been identified by combined analysis, it could be used to monitor residual disease during/after chemotherapy, by quantitative RT-PCR and/or FISH.
3.The study on local field potentials in rat's primary motor cortex during pressing paddle behavior.
Chinese Journal of Applied Physiology 2011;27(1):37-40
OBJECTIVEThe local field potential (LFP) is a summation of dendritic potentials. The main objective of the present work is to view the features of LFP in M1 during the experimental rat pushed a paddle with the right forelimb.
METHODSFour rats were trained to press a paddle with the right forelimb for water. Then Two bundle micro-electrode with four channels were implanted into the rat's left and right primary motor cortex (M1)(AP + 3.0 mm, ML +/- 1.6 mm, H-1.6 mm) with stereotaxic apparatus. After three days recovery, 8-channel Deep-EEG and the pulse signal of paddle pressed were recorded during the rats were in operant chamber, and at the same time, the behavior were also recorded with video recorder.
RESULTSThe LFP in left M1 were defined as the substance between two channel deep-EEG. It is found that low frequency, high amplitude signal appear aligned with the paddle pressed pulse signals. With threshold detect method, about 80% press-paddle behavior could be detected.
CONCLUSIONThe result indicates that LFPs in this position in M1 are relative to forelimb's movement, and a powerful brain-computer interface system maybe developed with the LFPs.
Animals ; Brain-Computer Interfaces ; Evoked Potentials, Motor ; physiology ; Male ; Microelectrodes ; Motor Cortex ; physiology ; Movement ; physiology ; Rats ; Rats, Sprague-Dawley
4.Preparation technology and dissolution research of Blumea volatile oil suppository.
Song WANG ; Yong-Heng ZHAO ; Yi-Sheng ZHOU ; Fang-Fang LI
China Journal of Chinese Materia Medica 2014;39(10):1805-1810
OBJECTIVETo research the preparation technology and dissolution of Blumea volatile oil suppository.
METHODIn order to establish the content determination and methodology inspection method of Blumea volatile oil plug, the extraction process of Blumea volatile oil was optimized by using orthogonal test. Optimization on the investigation to the suppository matrix by melting time, appearance and dissolution was carried on. The best prescription craft was determined by determining the best molding temperature, dosage of the matrix and complementary makings. The determination method of dissolution was established by investigating different dissolution method and its impact on the preparation of dissolution.
RESULTThe best conditions of steam distillation extracted Blumea volatile oil was as followed, the ratio of gardenia to liquor 1:6, 2.5% drug amount of sodium, 8 hours of extracting time. The optimum temperature for mold was 60-65 degrees C. Preparation technique of Blumea volatile oil suppository was stable, which after 45 minutes and 3 h in pH 4.5 PBS released at least 70% and 90%.
CONCLUSIONBlumea volatile oil suppository with rational prescription, simple preparation and good stability.
Asteraceae ; chemistry ; Chemistry, Pharmaceutical ; methods ; Distillation ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Solubility ; Temperature
5.Cluster Analysis of Medication Laws for Treating Coronary Heart Disease by Distinguished Veteran Doctors of Traditional Chinese Medicine.
Yi REN ; Zhi-qiang CHEN ; Min-zhou ZHANG ; Li-heng GUO ; De-ying HE
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(4):411-414
OBJECTIVETo provide inspiration and ideas for clinical treatment of coronary heart disease (CHD) by data mining technology based frequency analysis and cluster analysis of medical records, prescriptions and herbs in treating CHD by distinguished veteran doctors of traditional Chinese Dedicine (TCM).
METHODSTotally 386 medical cases were retrieved from Wanfang Data, Chinese Scientific Journals Database (VIP medical information resources system, China National Knowledge Infrastructure (CNKI), and Typical Collections of Medical Cases by Contemporary Distinguished Veteran Doctors of Traditional Chinese Medicine. They input into database trimmed after unified standard. Medication laws of CHD by distinguished veteran doctors of TCM were analyzed using frequency analysis and cluster analysis, and so on.
RESULTSDistinguished veteran doctors of TCM frequently used top ten herbs in treatment of C D as Salvia miltiorrhiz , Ligusticum wallichii, Trichosanthes kirilowi, Pinellia ternat, Angelica sinensis, Poria coco stragalu , Panax ginseng, Allium macrostemon, and Radix Ophiopogonis. Cluster analysis summarized that there were 16 herb pairs commonly used, 7drug assemblies consisting of 3 herbs and 5 drug assemblies consisting of multiple herbs.
CONCLUSIONSDistinguished veteran doctors of TCM mainly used herbs assemblies capable for invigorating Pi to resolve phlegm, and promoting qi and activating blood circulation in treating CHD. Meanwhile, they concurrently used herbs combination of nourishing Xin and tranquilization, and regulating yin and yang.
China ; Cluster Analysis ; Coronary Artery Disease ; therapy ; Data Mining ; Databases, Factual ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional
7.Construction and sequencing of recombinant lentiviral plasmid affecting human CCL20 gene by RNA interfering
Zhengxue DONG ; Daizhi PENG ; Jing LIU ; Xin ZHOU ; Yi TIAN ; Fang LI ; Quan YAN ; Heng LIN ; Yong WANG ; Guangqia ZHOU
Journal of Third Military Medical University 2003;0(10):-
Objective To design the small interference RNA (siRNA) specific to human CCL20 gene by RNA interfering technique, construct its recombinant lentiviral expression vectors, and identify these vectors by DNA sequencing. Methods According to Tuschl’s principle, the siRNA was designed and converted into cDNA of shRNA (small hairpin RNA) of siRNA for CCL20 gene. The cDNA was synthesized and inserted into plasmid pHSER-dsRNA-GFP-SIN which was linearized by restriction endonucleases SpeⅠ and SalⅠ. The recombinant plasmid was transformed into competent E. coli. DH5? cells. The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing. Results Two recombinant lentiviral plasmids of siRNA specific to CCL20 gene were constructed successfully. Their DNA sequence analysis completely coincided with their designed sequences. Conclusion Lentiviral vector-based siRNA expression plasmids against CCL20 gene have been successfully constructed and identified. They will be further used for interfering CCL20 mRNA transcription and lay the foundation for CCL20 gene modified human keratinocyte stem cells.
8.Interventional effects of donkey-hide glue reinforcing bone oral solution on the expression of growth factors during fracture healing
Heng LI ; Lin SHEN ; Liqin LI ; Yanping YANG ; Jing XIE ; Piqi ZHOU ; Yi DAI ; Furong LU ; Jialin WU
Chinese Journal of Tissue Engineering Research 2007;11(23):4634-4637
BACKGROUND:Donkey-hide glue reinforcing bone oral solution (DGRBOS) is effective on preventing and treating fracture, but the mechanism of harmacology is still not clear. Vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2) are important cytokines, which can promote blood vessel growth and osseous anabolism during fracture healing.OBJECTIVE: To investigate the effects of DGRBOS on expression of VEGF and FGF-2 in the process of fracture healing of SD rats' fracture of tibia in bony callus, and explore the mechanism of DGRBOS in the treatment of fracture.DESIGN: A completely randomized controlled study.SETTING: Department of Traumatic Orthopedics, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: Ninety female Sprague-Dawley (SD) rats, with a mean weight of (368±40) g, aged about 3 months, wereprovided by Center of Animal Experiment, Tongji Medical College, Huazhong University of Science and Technology.Experimental drug: DGRBOS was donated by Xinjiang Huashidan Pharmaceutical Co., Ltd. The qili linking bone pill,positive control drug, was purchased from Hunan Pharmaceutical Co., Ltd.METHODS: This experiment was carried out in the Laboratory for Bone Metabolism of Integration of Chinese and Western Medicine (Laboratory of Provincial Level), Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from June to November 2005. Transverse midshaft fractures were produced on the right tibia in these rats using three-point bending technique, and 90 rats were randomly divided into three experimental groups:the DGRBOS group (n =30): according to medicine conversional method between human being and animal, 2 mL DGRBOS was fed to every mouse by intragastric administration at 2 vices/day; the qili linking bone pill group (positive control group, n =30): the pills dissolved in distilled water at 225 g/L, and consequently were administered intragastrically into mice at 2 mL/vice and 2 vices/day; normal saline group (negative control group, n =30): normal saline was administered intragastrically into mice at the coordinative capacity and same frequences. Selecting 4, 7, 14,21 and 28 days during the experiment, the immunohistochemistry method was adopted to detect the change of expression of VEGF and FGF-2 through computing value of mean optical degree (MOD) and number of the positive cells.
9.CD11c+DCs promote the occurrence and development of psoriasis-like morbidity in K14-VEGF transgenic mice
Yi ZHANG ; Biao ZHOU ; Zheng-Hong GUO ; Hu LONG ; Heng-Xiu YAN
Basic & Clinical Medicine 2018;38(2):205-212
Objective To investigate the auxoaction and mechanism of CDllc+DCs on psoriasis. Methods CDllc+ DCs in the psoriasis-like lesions were confirmed by immunofluorescence double staining experiments. The CD1lc+ DCs were subcutaneously injected into the neck skin of transgenic mice without disease. The clinical features were observed and assessed with PASI score every day. The ear and back skin were checked by HE stained. At the same time, the T lymphocyte and DCs of bone marrow, spleen, submandibular lymph nodes and blood were analyzed by flow cytometry, and the T lymphocyte in the skin lesions were analyzed by immunofluorescence. The heart, liver, spleen, lungs and kindey were HE stained. Results In the psoriasis-like lesions, about 90% CDllc+ cells expressed CDllc. HE test showed that the thickness of the skin layer was significant different between the experimental group (ear: 29±4 μm; back: 25±3 μm) and the control group(ear: 11±2 μm; back: 9±1 μm) (P<0.01). CD3 + CD4+ T lymphocyte cells in the blood of the experimental mice (17.87%) were decreased significantly (P<0.01) compared with the control group mice (31.77%). The numbers of Thl and Th17 cells from bone marrow, spleen, submaxillary lymph nodes and blood, as the same as CD1lc+DCs from bone marrow and submaxillary lymph nodes, were decreased in the experimental mice compared with the control group mice (P<0.01). The number of CD4﹢T cells, CD8+T cells, IL-17a+cells and IFN-γ+cells in the skin lesions from the experimental group all were higher than that from the control group (P<0.01). And the vessels in the lesions of the experimental group were higher than that in the control group (P<0.01). Conclusions CD1 lc+CD1 lc+DCs may play an important role in the occurrence and development of psoriasis by increasing the T lymphocyte and the blood vessel in the skins of K14-VEGF transgenic mice.
10.Early use of high-potency antibiotics within short courses: a prospective clinical study
Xin-Zhou RONG ; Wen-Zhen ZHANG ; Jia-Liang REN ; Wei-Ming ZHOU ; Yong-Heng WU ; Lei YAN ; Yi-Ping ZHOU
Journal of Southern Medical University 2000;20(6):549-550
Objective To evaluate the effect of early and short-term use of high-potency antibiotics following extensive severe burn injury. Methods Seventeen severely burned patients hospitalized in the same period (Nov, 1998 to Oct, 2000) were randomly divided into 2 groups. Patients in group 1 received early treatment with imipenum and amikacin for 5 d, and the same treatment in group 2 was discontinued on day 15 postburn. The survival rate, blood bacterial culture, body temperature and white blood cell and platelet counts were compared between the 2 groups. Results All the 17 patients survived and all blood bacterial cultures were negative. No significant difference in the body temperature or white blood cell and platelet counts between the 2 groups was observed (P>0.05). Conclusion Early use of high-potency antibiotics at a short treatment course after extensive severe burn is effective to prevent infection and reduce the cost.