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.Protection and mechanism of shenqi compound for diabetic angiopathy model rats.
Yong-He HU ; Jun HOU ; De-Zhi ZHENG ; Dan-Dan LI ; Xin-Zhong HAO ; Chun-Guang XIE ; Lian DU ; Qing NI ; Yi SHEN ; Jing LI
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(9):1078-1085
OBJECTIVETo investigate the protective effect and mechanism of Shenqi Compound on diabetic angiopathy modeled rats.
METHODSTotally 18 SD rats were randomized into 3 groups, i.e., the normal control group, the diabetic mellitus (DM) group, and Shenqi Compound group, 6 in each group. The DM rat model was established by feeding high-fat diet (to induce hyperlipidemia) +intraperitoneal injection of small dose streptozotocin (STZ). Shenqi Compound was given to rats in the Shenqi Compound group at the daily dose of 2 g/kg. Equal volume of normal saline was given to rats in the model group and the normal control group by gastrogavage. All treatment was lasted for 12 weeks. Then 2-D and ultrasonic integrated backscatter technique were used to evaluate structural and functional changes of abdominal aorta in the progression of diabetic macroangiopathy. The fibrosis degree of the aorta vessel and myocardium capillaries were observed by using HE and Masson trichrome staining. The tension of the aortic vascular ring was determined. The transforming growth factor beta (TGF-beta) mRNA expression was detected by real time PCR (RT-PCR). The protein expression of TGF-beta, collagen I, collagen III, connective tissue growth factor (CTGF), and phosphorylation P38 MAPK were detected by Western blot.
RESULTSCompared with the normal control group, abdominal aortic systolic inner diameter, diastolic inner diameter, Peterson elastic modulus, stiffness index, and backscatter integral significantly increased; the rangeability of integral backscatter and the extension coefficient of cross section significantly decreased in the DM group (all P < 0.05). After 12 weeks aforesaid indices were obviously improved in the Shenqi Compound group (P < 0.05). Results of HE and Masson staining showed that the fibrosis degree of the aorta vessel and myocardium capillaries was obviously alleviated in rats of the Shenqi Compound group (P < 0.05). Results of the aortic vascular ring tension showed that acetylcholine induced vasodilatation and maximum diastolic percent were obviously elevated in the Shenqi Compound group (P < 0.05). Compared with the normal control group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all significantly increased in the DM group (P < 0.05). Compared with the DM group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all decreased (P < 0.05).
CONCLUSIONSShenqi Compound could effectively improve the arterial function in diabetic marcoangiopathy and microvascular dysfunction. The mechanism might be due to the down-regulating the expression of TGF-beta, and further suppressing the phosphorylation of P38 MAPK, reducing the synthesis of collagen I and collagen III, therefore, ameliorating arterial and myocardial interstitial fibrosis.
Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Angiopathies ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism
9.Prolonged duration of the routine pegylated-interferon alfa-2a therapy produces superior virological response in HBeAg-positive chronic hepatitis B patients: a single-center cohort study.
Yue-yong ZHU ; Yin-lian WU ; Jing CHEN ; Qi ZHENG ; Jing DONG ; Jia-ji JIANG
Chinese Journal of Hepatology 2012;20(10):737-741
OBJECTIVEHepatitis B e antigen (HBeAg) seroconversion and/or hepatitis B surface antigen (HBsAg) clearance are considered as good prognostic indicators of treatment outcome in HBeAg-positive chronic hepatitis B (CHB) patients. While a sustained virological response (SVR) can be achieved by a finite 48-week course of pegylated-interferon alfa-2a (Peg-IFNalpha-2a), it has been suggested that longer-term treatment can improve the rate of SVR. Therefore, the aim of this study was to compare the effects of prolonged and routine Peg-IFNa-2a therapy in patients with HBeAg-positive CHB.
METHODSEighty-six consecutive patients diagnosed with HBeAg-positive CHB at our hospital between September 2006 and October 2009 were enrolled in the study. The patients were randomly assigned to receive Peg-IFNa-2a (180 mug once weekly) for either 48 weeks (routine therapy group, n = 53) or 72 weeks (prolonged therapy group, n = 33). Serum samples were collected from each patient every three months until the end of the 24-week follow-up, and standard viral and biochemical tests were carried out. Relapse was defined as HBV DNA concentrations more than 105 copies/mL or an HBeAg-positive test at the end of the 24-week follow-up. Chi-squared test and the t-test were used to determine the significance of intergroup differences. Logistic regression analysis was employed to determine the correlation of outcome parameters to treatment duration, expressed as odds ratio (OR) with 95% confidence interval (CI).
RESULTSThe two treatment groups were similar at baseline (pre-treatment) in demographic data, sex ratio, age, alanine aminotransferase (ALT) level, HBV DNA load, and semi-quantitative level of HBeAg (s/co) (all, P more than 0.05). At the end of the 24-week follow-up, there were significant differences between the 48-week treatment group and the 72-week treatment group in patients with HBV DNA negativity (62.3% vs. 97.0%, x2 = 13.273, P = 0.000), HBeAg seroconversion (39.6% vs. 57.6%, x2 = 6.765, P = 0.009), HBsAg clearance (15.1% vs. 36.4%, x2 = 5.155, P = 0.023), and relapse (58.5% vs. 33.3%, x2 = 6.713, P = 0.010). Logistic regression analysis indicated that therapy duration was correlated to HBeAg clearance (OR = 3.702, 95% CI: 1.225 to 11.188) and male sex (OR = 3.005, 95% CI: 1.038 to 8.696) but not to HBeAg level at baseline (OR = 0.999, 95% CI: 0.998 to 1.000) or age (OR = 0.902, 95% CI: 0.839 to 0.970).
CONCLUSIONIn this single-center cohort study, superior therapeutic benefit was achieved by extending the Peg-IFNa-2a therapy out to 72 weeks for patients with HBeAg-positive CHB. The prolonged duration therapy produced a higher HBsAg loss ratio, HBeAg seroconversion ratio, HBV DNA negativity ratio, and a lower relapse ratio. Furthermore, HBeAg clearance was positively correlated with duration and male sex.
Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Treatment Outcome ; Young Adult
10.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