2.Construction and expression of recombinant cecropin B-binding site of luteinizing hormone releasing hormone gene and its anticancer function
Xiao-Yong LI ; He-Lian LI ; Gui-Ying ZHENG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(07):-
Objective To construct and express recombinant cecropin B-binding site of luteinizing hormone releasing hormone(CB-LHRH')gene,and to evaluate the anticancer function of CB-LHRH' on human ovarian cancer cell line SKOV3 and human endometrial cancer cell line HEC-1B.Methods The sequence of the cDNA encoding CB-LHRH' was designed,artificially synthesized,verified by DNA sequence analysis and expressed by Bac-to-Bac baculovirus expression system.The expression of CB-LHRH' proteins were identified by western dot blot using rabbit polyclonal antibody against LHRH as the primary antibody.To determine the anticancer effects of the CB-LHRH' protein,ovarian cancer cell line SKOV3 and endometrial adenocarcinoma cell line HEC-1B were treated by different doses of the CB-LHRH' protein.Cell growth inhibition assay was performed using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)5[(phenylamino) carbonyl]-2H-tetrazolium hydroxide(XTT)kit at different times,and cell morphologic changes were observed under the inverted microscope.Results The inhibitory rate of proliferation by CB-LHRH' increased with the increase of dose and time respectively:SKOV3 cell,from(5.03?0.08)% to(53.24 ?1.22)%;HEC-1B cell,from(5.13?0.37)% to(56.16?1.08)%.The inhibitory effect on HEC-1B cell was stronger than that on SKOV3 cell(P
3.Reconstruction intramedullary nailing for treatment of ispsilateral femoral neck and shaft fractures.
Hao WANG ; Lian-hua LI ; Zhi LIU ; Jian-zheng ZHANG ; Yong-zhi GUO
China Journal of Orthopaedics and Traumatology 2015;28(9):808-810
OBJECTIVETo evaluate the curative effect and surigical skills of reconstruction intramedullary nailing in the treatment of ipsilateral femoral neck and shaft fractures.
METHODSFrom January 2007 to January 2013, 13 patients with ipsilateral femoral neck and shaft fractures were treated by reconstruction intramedullary nailing including 11 males and 2 females with an average age of 38.9 yeas old ranging from 25 to 53 years old. For femoral neck fractures,10 cases were basilar neck fracture, 3 were transcervical fractures and according to Garden classification,10 were type II, 2 were type III and 1 was type IV. For feormal shaft fracture, 5 were type I, 4 were type II, 3 were type III and 1 was type IV according to Winquist classification. The location of all 13 feormal shaft fractures were all in the uper 3/4 of the feormal shaft. Complications and postoperative function were recorded and evaluated.
RESULTSEleven patients were followed up for 23.45 months (12 to 36 months). Time from injury to operation was 5.9 days in average. Malunion of femoral neck fracture happened in 1 patient, nonunion of femoral shaft fracture happened in 2 patients. NO femoral head osteonecrosis, infection and malunion of femoral shaft fracture happened. According to Friedman-Wyman classification, 8 cases get good result, 2 get faii and 1 get poor.
CONCLUSIONReconstruction intramedullary nailing is a good choice for the ipsilateral femoral neck and shaft fractures with the advantages of less soft-tissue trauma and complications.
Adult ; Female ; Femoral Fractures ; surgery ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged
4.Influence of cryoprotectant and cooling rate in vitrification method on the spindles of rabbit oocytes
Xue-Yong CAI ; Gui-An CHEN ; Ying LIAN ; Xiao-Ying ZHENG ; Hong-Mei PENG ;
Journal of Peking University(Health Sciences) 2003;0(06):-
SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.
5.Inhibitory effects of emodin on drug-resistant HL-60/ADR cell proliferation and its induction of apoptosis.
Ying-Yu CHEN ; He-Yong ZHENG ; Jian-Da HU ; Zhi-Hong ZHENG ; Jing ZHENG ; Xiao-Lan LIAN ; Lian-Huang LÜ
Journal of Experimental Hematology 2007;15(5):955-960
The study was aimed to investigate the effects of emodin on the proliferation and apoptosis of adriamycin-resistant HL-60/ADR cells, and to explore the underlying mechanism. The cell viability and colony formation were detected by MTT assay and colony formation assay respectively. Apoptotic cells were tested by means of cell cycle analysis, mitochondrial transmembrane potential levels, caspase-3 activity detection, Annexin V FITC/PI staining and TUNEL labeling. RT-PCR was used to analyze the bcl-2 and c-myc mRNA expressions. The protein expressions of Bcl-2, c-Myc and caspase-3 precursor were determined by Western blot. The results showed that HL-60/ADR cell growth was significantly inhibited by emodin in dose and time dependent manners. Cell colony formation obviously decreased with IC50 5.79 micromol/L. G0/G1 phase cell population increased while G2/M phase cells decreased in 40 and 80 micromol/L groups compared with control group (p < 0.01), and no significant difference of cell cycle was observed in 20 micromol/L group (p > 0.05). The typical hypo-diploid peak (apoptotic peak) appeared in each dose group. The levels of mitochondrial transmembrane potential of HL-60/ADR cells decreased and caspase-3 activity increased when incubated with emodin for 12 and 24 hours respectively. Apoptosis occurred in a dose-dependent manner, and its earlier and later stages were identified by Annexin-V FITC/PI staining and TUNEL labeling methods respectively. The expressions of bcl-2, c-myc mRNA and Bcl-2, c-Myc, caspase-3 precursor protein were all down-regulated in a time-dependent manner after treatment with emodin at different times. It is concluded that emodin efficiently inhibits growth and induces apoptosis on HL-60/ADR cells, which may be related with the down-regulation of mitochondrial transmembrane potential and expressions of bcl-2 and c-myc, as well as up-regulation of caspase-3 activity.
Apoptosis
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drug effects
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Caspase 3
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metabolism
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Cell Proliferation
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drug effects
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Doxorubicin
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pharmacology
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Drug Resistance, Neoplasm
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drug effects
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Emodin
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pharmacology
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HL-60 Cells
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Humans
6.A Corpus-Based Stylistic Analysis on Structured Abstract of Literature Review Papers
lan-ying, WANG ; lian-zhu, WANG ; jia-qin, WANG ; rui-jun, ZHANG ; wen-ming, YONG ; hui-shao, ZHENG
Journal of Applied Clinical Pediatrics 2006;0(23):-
To introduce the writing techniques for structured Abstract of literature review papers based on the knowledge of corpus linguistics,so as to summarize the stylistic,structural,syntactic and lexical characteristics,objectively and systematically.This research will also faci-litate the authors and translators,who have difficulties in writing structured English Abstract,to publish their articles via mastering its stylistic characteristics,writing format and language.
7.Transcriptome profiling and analysis of Panax japonicus var. major.
Shao-peng ZHANG ; Jian JIN ; Bing-xiong HU ; Ya-yun WU ; Qi YAN ; Wan-yong ZENG ; Yong-lian ZHENG ; Zhang XI-FENG ; Ping CHEN
China Journal of Chinese Materia Medica 2015;40(11):2084-2089
The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling
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Panax
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genetics
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Saponins
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biosynthesis
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Sequence Analysis, RNA
8.Effects of bushen jiannao recipe on the content of acetylcholine and the hippocampal ERK1 and ERK2 protein expressions of vascular dementia rats.
Yong-Hui LIU ; Shao-Wei LI ; Qing-Lian ZHENG
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(4):504-509
OBJECTIVETo explore the effects of Bushen Jiannao Recipe (BJR) on the content of acetylcholine (Ach) and ERK1 and ERK2 protein expressions in the hippocampal CA1 region of vascular dementia (VD) rats, and to explore its possible mechanisms for treating VD.
METHODSEighty-three rats were selected. The VD model was established by permanent bilateral occlusion of both common carotid arteries (2-VO). Then the modeled rats were randomly divided into 5 groups, i. e., the memory deficit model group, the donepezil group, and the positive drug control groups [including high (n = 13), middle (n = 13), and low (n = 12) dose BJR group]. Besides, another 13 rats were chosen as the sham-operative group. The distilled water was given by gastrogavage to rats in the sham-operative group and the memory deficit model group (5 mL/kg). The donepezil hydrochloride suspension was given to rats in the donepezil group by gastrogavage (0.52 mg/kg). High (56 g/kg), middle (28 g/kg), and low (14 g/kg) dose of BJR were respectively given to rats in the other three groups. After 30 days of intervention, the escape latency period and platform crossing times were determined using Morris water maze experiment. The contents of Ach in the hippocampus and cortex were determined using colorimetry. The expressions of ERK1 and ERK2 in the CA1 region of the hippocampus were detected using immunohistochemical assay.
RESULTSThe average escape latency of intervened rats showed an overall decreasing trend. From the third to the fifth day, the escape latency period was prolonged, the platform crossing times were reduced, the contents of Ach in the cortex and the hippocampus were lowered, the numbers of positive stained neuron of ERK1 and ERK2 in the hippocampus CA1 region were reduced, showing statistical difference when compared with the sham-operative group (P<0.01). Compared with the model group, the 4th day escape latency of the donepezil group and the high dose BJR group was shortened. The escape latency was shortened, and the platform crossing times, and the numbers of positive stained neuron of ERK1 and ERK2 in hippocampus CA1 region increased on the fifth day. The contents of Ach in the cortex and the hippocampus increased with statistical difference (P<0.05). Compared with the low dose BJR group, the 4th- and 5th-day latency period were shortened, the positive numbers of ERK1 and ERK2 in the hippocampus CA1 region increased in the high dose BJR group with statistical difference (P<0.05). Compared with the donepezil group, the Ach content in the cortex and the hippocampus of the middle and low dose BJR groups decreased (P<0.05).
CONCLUSIONSBJR could obviously improve the function of learning and memory of VD rats. Its mechanisms might be associated with its actions in enhancing Ach contents of the cortex and the hippocampus, and promoting the protein expressions of ERK1 and ERK2 in the hippocampus CA1 region.
Acetylcholine ; metabolism ; Animals ; CA1 Region, Hippocampal ; metabolism ; Dementia, Vascular ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; MAP Kinase Signaling System ; drug effects ; Male ; Maze Learning ; drug effects ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Rats ; Rats, Sprague-Dawley
9.Tramadol inhibits c-fos expression in spinal cord dorsal horn and serum IL-6 levels induced by plantar incision in rats.
Yong-min LIU ; Sheng-mei ZHU ; Kui-rong WANG ; Qing-lian CHEN ; Yue-ying ZHENG
Journal of Zhejiang University. Medical sciences 2009;38(4):392-398
OBJECTIVETo investigate effect of tramadol on c-fos expression in spinal cord dorsal horn and serum IL-6 levels induced by plantar incision in rats.
METHODSThe Brennan pain model was induced by incision on the planter surface of left hind paw in rats. Forty-eight rats were randomly divided into six groups: Sham group (Group C), control group (Group I,pretreatment with saline 5 ml), three tramadol pretreatment groups (Group T1, T10 and T20,pretreated with 1 mg/kg, 10 mg/kg and 20 mg/kg tramadol, respectively) and one tramadol treatment group (Group PT10, treated with tramadol 10 mg/kg immediately after operation). Pain behavior was assessed by withdrawal threshold to von Frey filament stimulation intensity, response latency of the hind paw to radiant thermal and a cumulative pain score 2 h after incision. Fos-positive neurons in spinal cord were identified by the immunohistochemical technique. Serum IL-6 levels were measured by enzyme-linked immunosorbent assay (ELISA).
RESULTSWithdrawIal threshold to von Frey filament stimulation intensity and response latency of the hind paw to radiant thermal in Group I were significantly lower than those in Group C (P<0.01). Cumulative pain score in Group I was significantly higher than that in Group C (P<0.01). In Groups of T10 and T20, withdrawal threshold to von Frey filament stimulation intensity and response latency of the hind paw to radiant thermal were significantly higher than those in Group I (P<0.01), cumulative pain score was significantly lower than that in Group I in a dose-dependent manner (P<0.01), and were also those in Group PT10. The greatest density of Fos-positive neurons was located in lamine I-II in Group I. Serum IL-6 levels were significantly elevated in Group I. Pretreatment with tramadol showed a dose-depended inhibitory effect on c-fos expression and serum IL-6 production,but not in Group T1. Administration of tramadol postoperatively also suppressed the c-fos expression and serum IL-6 production as showed in PT10 but were weaker than those in Group T10.
CONCLUSIONPretreatment with tramadol can produce dose-dependent inhibitory effect on c-fos expression in spinal cord dorsal horn and then suppress the inflammatory response to the trauma.
Analgesics, Opioid ; pharmacology ; therapeutic use ; Animals ; Interleukin-6 ; blood ; Male ; Pain Threshold ; drug effects ; Pain, Postoperative ; drug therapy ; metabolism ; Posterior Horn Cells ; drug effects ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tramadol ; pharmacology ; therapeutic use
10.Mutation Analysis of CIB2 196C>T 272T>C and 297C>G in Patients with Non-syndromic Hearing Impairment
Qi LI ; Jun-Dong HE ; Jin-Li WANG ; Yong-Qin ZHENG ; Rong-Xia ZUO ; Ya-Lian SA
Journal of Kunming Medical University 2018;39(5):7-10
Objective To investigate the gene mutations of calcium-and integrin-binding protein 2 (CIB2) 196C>T, 272T > C and 297C > G carried by students with non-syndromic hearing impairment from special educational schools in Yunnan Province. Methods The experimental group included 337 students with non-syndromic hearing impairment who failed to carry deafness gene with GJB2 (35 del G, 176_191 del 16,235delC, 299_300 del AT), GJB3 (C538T,G547A), mtDNA 12S rRNA (A1555G, C1494T), and SLC26A4 (IVS7_2A>G, A2168G) . The control group consisted with 150 healthy people. Genomic DNA was isolated from peripheral blood with EDTA anti-coagulate. The subject's DNA fragments including CIB2 196C>T, 272T > C and 297C> G were amplified by polymerase chain reaction (PCR), and subsequently analyzed by direct sequencing to identify deafness-associated mutations. Results Both in the experimental group and control group, we failed to find the mutation of CIB2 196C>T, 272T>C and 297C>G in all individuals. Conclusion Mutations in CIB2 gene 196C>T, 272T>C and 297C>G are not a frequent cause of non-syndromic hearing loss among deaf people in Yunnan province. It provided important information for deafness with formulating landscape of gene screening in this region.