2.Heterozygous genotypes and molecular characteristics of Organophosphorus resistance associated esterase B2 genes of Culex pipiens complex.
Yu KOU ; Xin-fen YU ; Rong YE ; Jin-cao PAN ; Feng CUI ; Chuan-ling QIAO
Chinese Journal of Preventive Medicine 2009;43(5):390-394
OBJECTIVETo investigate the heterozygous genotype and molecular characteristics of Organophosphorus resistance associated with heterozygous Estbeta2 of esterase B2 gene from natural population of Culex pipiens complex.
METHODSGenomic DNA was extracted from natural populations of Culex pipiens complex in Hangzhou. The PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase gene. Estbeta2 of esterase B2 gene was identified by PCR-RFLP, and the genotyping for heterozygous Estbeta2 was carried out after restriction enzyme digesting by Bfm I endonuclease.
RESULTSThe DNA was isolated from 207 Culex pipiens respectively, while 156 PCR samples showed positive and the positive rate was 75.36% (156/207). The PCR-RFLP assay of esterase B2 gene revealed that the Estbeta2 was accounted about 28.20% (44/156) in 156 positive samples. There were two genotypes identified, namely homozygous Estbeta2 (90.90%, 30/33) and heterozygous Estbeta2 (9%, 3/33), heterozygous Estbeta2 was in existence of a hybrid form as which combined with Estbeta2 and a subtype (Estbeta2/Estbeta2(1)).
CONCLUSIONHeterozygous Estbeta2 of Organophosphorus resistance associated with esterase genotype was determined in natural population of Culex pipiens, and a genotyping method was established.
Animals ; Culex ; enzymology ; genetics ; Genes, Insect ; Genotype ; Heterozygote ; Insecticide Resistance ; genetics ; Insecticides ; pharmacology ; Organophosphorus Compounds ; pharmacology ; Phenotype ; Serine Endopeptidases ; genetics
3.Cloning and sequence analysis of MYB transcriptional regulator SmP gene of Saussurea medusa Maxim.
Zhi-Ping JIN ; De-Xiu ZHAO ; Chuan-Ling QIAO ; Wen-Quan QU ; Ya-Qiong CHEN ; Chun-Xiang FU
Chinese Journal of Biotechnology 2003;19(3):368-371
A full-length cDNA encoding a MYB-related regulatory gene was isolated from a cDNA library prepared from mRNAs of the red line callus of S. medusa by TD-PCR. The cDNA, designated SmP, is 969 nucleotides long and has an open reading frame of 771 bp with a deduced amino acid sequence of 256 residues. The putative protein of SmP has two typical conversed R2R3-Myb DNA-binding domains in N-terminal and displays a rather high degree of similarity to OsMYB from rice and LBMI from tobacco, showing 73% and 70% identity within the DNA-binding domains. However, the C-terminal domain of the SmP protein does not show obvious similarity to any other known protein sequence. It is rich in hydrophilic amino acids, especially in serine residues (18.38%), partly organized in homopolymeric stretches, a feature often found in activation domain of transcription factors.
Amino Acid Sequence
;
Cloning, Molecular
;
Molecular Sequence Data
;
Phylogeny
;
Plant Proteins
;
chemistry
;
classification
;
genetics
;
metabolism
;
Polymerase Chain Reaction
;
Saussurea
;
classification
;
genetics
;
metabolism
;
Sequence Homology, Amino Acid
;
Transcription Factors
;
chemistry
;
genetics
;
metabolism
4.Genotypes, allele frequencies and dynamic distribution on resistance-associated esterase genes of Culex pipiens complex in Hangzhou.
Yu KOU ; Chuan-Ling QIAO ; Jin-Cao PAN ; Feng CUI ; Rong YE ; Xin-Fen YU
Chinese Journal of Preventive Medicine 2007;41(6):483-486
OBJECTIVETo investigate the genotypes , allele frequencies and dynamic distribution on resistance associated esterase genes of Culex pipiens complex in Hangzhou.
METHODSThe PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase genes, and dynamic surveillance on frequencies of the resistance associated esterase gene of natural population of Culex pipiens complex in Hangzhou during 2003-2005, and phenotype of the resistance associated esterase genes were detected by esterase starch gel electrophoresis technique.
RESULTSThe PCR-RFLP assay of esterase allele genes for three consecutive years disclosed four esterase genotypes, namely, the world-wide highly active homozygous Est beta 1(1) (50%-54%), homozygous Est beta 2 (29%-34%), heterozygous Est beta 1(1)/beta 2 (5%-10%) and Est beta N (3.13%) of a new homozygous genotype. The research of the resistance associated esterase genes phenotype in natural population of Culex pipiens complex in Hangzhou in 2005 with esterase starch gel electrophoresis technique revealed four major types, namely, Est beta 1(1) (61%), Est alpha 2/beta 2 (12%), Est alpha 8/beta 8 (7%) and sensitive phenotype (29%).
CONCLUSIONThere should be various resistance associated esterase genotypes in natural population of Culex pipiens complex in Hangzhou. During the period of 2003-2005, Est beta 1(1) was the major type; Est alpha 2/beta 2 was the second. Est beta N was a new esterase genotype detected in 2005 only with a mere percentage of 3.13%. As for its resistance to the new insecticide, a follow-up study should be needed. The molecular typing of the amplified esterase gene should be consistent with the resistance associated esterase genes phenotype.
Alleles ; Animals ; China ; Culex ; genetics ; physiology ; Esterases ; analysis ; genetics ; Gene Frequency ; Genotype ; Insecticide Resistance ; genetics ; Phenotype
5.Clinical analysis of autoimmune hemolytic anemia after allogeneic hematopoietic stem cell transplantation in thalassemia major.
Zhong Ming ZHANG ; Yong Rong LAI ; Qiao Chuan LI ; Lin LUO ; Rong Rong LIU ; Ling Ling SHI ; Lian Jin LIU
Chinese Journal of Hematology 2018;39(11):908-911
Objective: To explore the diagnosis, treatment and prognosis of autoimmune hemolytic anemia (AIHA) after allo-HSCT in patients with thalassemia major (TM). Methods: A retrospective analysis of AIHA status after allo-HSCT in 291 TM patients from July 2007 to December 2017 was conducted. Results: Five of the 291 TM patients (1.72%) were diagnosed with post-transplant AIHA. The median time of AIHA was 7 (5-12) months after HSCT. All post-transplant AIHA patients were positive in direct and indirect Coombs test, the main clinical manifestations were dizziness, fatigue, pale complexion, skin and sclera yellow, and soy sauce urine. The incidence of AIHA was higher after unrelated donor transplantation (6.36%, 4/63) compared with that of sibling donor transplantation (0.43%, 1/228). One patient who received only prednison was dead. Four patients who received rituximab combined with prednisolone were alive, Coombs test in two of them were negative. Conclusions: AIHA after allo-HSCT developed in 1.72% patients with TM. Monitoring of Coombs test was important for diagnosis of post-transplant AIHA. The incidence of post-transplant AIHA was higher in unrelated donors compared with that of sibling donors transplantation. Treatment of rituximab combined glucocorticoid was effective strategy for post-transplant AIHA.
Anemia, Hemolytic, Autoimmune
;
Coombs Test
;
Hematopoietic Stem Cell Transplantation
;
Humans
;
Retrospective Studies
;
beta-Thalassemia
6.Cardiac protective effects of transcutaneous electrical acupoint stimulation combined with general anesthesia for controlled hypotension.
Jian-Qiao FANG ; Chuan-Long ZHOU ; Xiao-Mei SHAO ; Xiao-Qing GUO ; Le-Le ZHANG ; Ling JIN
Chinese Acupuncture & Moxibustion 2011;31(7):625-629
OBJECTIVETo explore the protective effect and the mechanism of controlled hypotension induced by transcutaneous electrical acupoint stimulation (TEAS) combined with general anesthesia.
METHODSSixteen male Beagles were randomly divided into a group of controlled hypotension induced by simple general anesthesia (control group) and a group of controlled hypotension induced by TEAS combined with general anesthesia (observation group). All the animals were administered with combination of Isoflurane and Sodium Nitroprusside (SNP) for controlled hypotension without TEAS until the arterial pressure was lowered to 30% basic mean arterial pressure (MAP) for 60 min. In the observation group, TEAS (2 Hz/100 Hz, 3-5 mA) was applied to "Hegu" (LI 4) "Zusanli" (ST 36), "Sanyinjiao" (SP 6) and "Quchi" (LI 11) from the beginning of physiological conditions stability to the end of maintained low MAP for 60 min, but there was no TEAS in control group. The changes of MAP, the left intraventricular pressure (LIVP), T wave and ST-T segment of II lead electrocardiogram (ECG) were monitored with the physiological signal acquisition system, and myocardial apoptosis was detected by TUNEL method.
RESULTSAll the animals could maintain stable required low blood pressure. At one hour after cease of controlled hypotension, MAP of (109.56 +/- 6.14) mmHg returned to the basic level in the observation group, while MAP of (84.91 +/- 6.36) mmHg was still lower than its basic MAP of (111.02 +/- 4.15) mmHg in the control group (P < 0.05), showing significant difference in MAP between the two groups (P < 0.05). -dp/dtmax of (3156.32 +/- 332.82) mmHg/s showed significant lower than its basic value of (4585.33 +/- 638.55) mmHg/s when blood pressure increased for 1 h in the control group (P < 0.05), but there was no difference in the observation group. When the objective low MAP maintaining for 60 min the ST segment was decreased significantly in the control group (P < 0.05), but there was no difference in the observation group. The numbers of positive apoptosis cardiocytes in the observation group were (24.67 +/- 2.45) cells/mm2, which were significantly fewer than (37.89 +/- 1.90) cells/mm2 in the control group (P < 0. 05).
CONCLUSIONSTEAS combined with general anesthesia for controlled hypotension can significantly shorten restoration time of MAP, help to improve myocardial ischemia and the cardiac functional recovery and reduce myocardial apoptosis so as to produce myocardial protection.
Acupuncture Points ; Anesthesia, General ; Anesthetics, General ; pharmacology ; Animals ; Blood Pressure ; drug effects ; Dogs ; Heart ; drug effects ; physiology ; Humans ; Hypotension, Controlled ; Male ; Models, Animal ; Random Allocation ; Transcutaneous Electric Nerve Stimulation
7.Effects of stromal cell-derived factor 1 and platelet factor 4 on the adhesion characteristics and chemotactic function of ex vivo expanded umbilical cord blood CD34+ cells.
Qiao-Chuan LI ; Yun-Tao LI ; Heng-Xing MENG ; Ya-Fei WANG ; Chang-Chun WAN ; Xin LI ; Wei GE ; Qian LI ; Jun-Ling HAN ; Lu-Gui QIU
Journal of Experimental Hematology 2006;14(1):83-88
To investigate the effects of stromal cell-derived factor 1 (SDF-1) and platelet factor 4 (PF4) on the homing-related function of expanded ex vivo umbilical cord blood CD34(+) cells, purified cord blood CD34(+) cells were cultured in serum-free medium containing a HGF combination of FL + SCF + TPO (FST) with either 100 ng/ml SDF-1 alone, 100 ng/ml PF4 alone, or both of these 2 cytokines. The expansion rate of CD34(+) cells, colony formation, homing-related functions including expression of homing-related adhesion molecules of expanded CD34(+) cell, adhesion activity and chemotactic function of the re-selected expanded CD34(+) cells were evaluated at different time points. The results showed that expansion rate of CD34(+) cells and expansion multiple of CFU in SDF-1 groups were higher than those in control. The expression of CD49e on the expanded CD34(+) cells was remarkable up-regulated, in contrast, expression of CXCR-4 on the expanded CD34(+) cells was remarkable down-regulated in SDF-1 groups. The expression of CD49e, CD54 and CXCR-4 on the expanded CD34(+) cells were remarkably up-regulated in the PF4 groups. In all the SDF-1 group, PF4 group and SDF-1 plus PF4 group, the ability of expanded CD34(+) cells adhering to fibronectin layer were higher than those in the control on day 10. Spontaneous migration rate of expanded CD34(+) cells in SDF-1 groups were higher than those in control, while SDF-1-induced migration rate were lower than those in control on day 10. SDF-1-induced migration rate in PF4 groups were higher than those in control on day 10. Spontaneous and SDF-1-induced migration rate of expanded CD34(+) cells in the SDF-1 plus PF4 groups were higher than those in control on day 10. It is concluded that, SDF-1 and PF4 can up-regulate expression of adhesion molecules on expanded CD34(+) cells, and retain the adherent and migration ability of expanded CD34(+) cells, which is helpful for the homing of expanded CD34(+) cells. In short, SDF-1 and PF4 are helpful for the homing-related function of the expanded UCB HSPC.
Antigens, CD34
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blood
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immunology
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Cell Adhesion
;
drug effects
;
Cells, Cultured
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Chemokine CXCL12
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Chemokines, CXC
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pharmacology
;
Chemotaxis
;
immunology
;
physiology
;
Culture Media, Serum-Free
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Fetal Blood
;
cytology
;
immunology
;
Hematopoietic Stem Cells
;
cytology
;
drug effects
;
Humans
;
Platelet Factor 4
;
pharmacology
8.Time-dependent expression of PEDF and VEGF in blood serum and retina of rats with oxygen-induced retinopathy.
Chun-tao LEI ; Xiao-ling WU ; Jie PENG ; Xiao-feng CHEN ; Li-feng QIAO ; Ying-chuan FAN ; Jian-bin HU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(1):135-139
The effects of the balance changes of pigment epithelium growth factor (PEDF) and vascular endothelial growth factor (VEGF) in whole-body and retinal tissue on rats with oxygen-induced retinopathy were investigated. Forty-eight neonatal SD rats at the age of 7 days were randomly divided into 4 groups. The neonatal rats in experimental groups were exposed to 75% to 80% oxygen for 5 days and then to normal air, and those in control groups were kept feeding in normal air. At the age of 17 and 22 days, all the neonatal rats received retina angiography with FITC-dextran and the pathological changes of retinal vessels and perfusion were observed. HE staining of the tissue section and the number counting of endothelial cells extending beyond the inner limiting membrane were performed to evaluate the endothelial proliferation. Immunohistochemistry was applied to detect the expression of PEDF and VEGF in retinal tissue, and ELISA to detect their expression in serum. A hypoxic-ischemic proliferation of retina and more endothelial cells extending beyond the inner limiting membrane were found in the neonatal rats in both experimental groups of 17-day old and 22-day old as compared with those in control group with the difference being statistically significant (P<0.01). VEGF staining of the rats in the 17-day old experimental group was significantly stronger, with an increasing positive rate, than that of the rats in the 17-day old control group (P<0.01). PEDF staining of the rats of 22 days old was weaker than that of the rats of 17 days old in the experimental groups (P<0.01). There was no significant difference in serum VEGF concentration among all groups (P>0.05). The serum PEDF concentration in the rats of 17 days old in experimental group was decreased significantly as compared with that in the rats of 17 days old in control group (P<0.01), and in experimental groups, the serum PEDF concentration of the rats of 22 days old was increased as compared with that of the rats of 17 days old (P<0.01). In conclusion, the obviously decreased serum PEDF concentration and the abnormal enhanced expression of VEGF density in local retinal tissue broke down the balance of PEDF/VEGF in whole-body or local tissues, which might play an important role in retinal vascular proliferation.
Animals
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Eye Proteins
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blood
;
metabolism
;
Nerve Growth Factors
;
blood
;
metabolism
;
Oxygen
;
adverse effects
;
Rats
;
Rats, Sprague-Dawley
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Retina
;
metabolism
;
Retinal Diseases
;
etiology
;
metabolism
;
Serpins
;
blood
;
metabolism
;
Time and Motion Studies
;
Vascular Endothelial Growth Factor A
;
blood
;
metabolism
9.Effect of the smoking cessation services in the out-patient department for patients with coronary heart disease.
Jing ZHAO ; Jian-jun GOU ; Da-yi HU ; Rong-jing DING ; Xiao-jun YU ; Fei HE ; Long WANG ; Ping ZHANG ; Xue-bin LI ; Ji-hong GUO ; Wen-ling LIU ; Cui-lan LI ; Lei LI ; Chuan-yu GAO ; Luo-sha ZHAO ; Ying-Jie CHU ; Zhen-wen HUANG ; Jing-han WEI ; Yi-bo WANG ; Xiao-feng ZHUANG ; Wei QIAO
Chinese Journal of Cardiology 2013;41(12):1000-1005
OBJECTIVETo evaluate the effects and clinical prognosis of out-patient department-based smoking cessation services for coronary heart disease (CHD) patients.
METHODSA total of 140 smoking patients diagnosed with coronary heart disease in our cardiovascular department were randomly divided into the intensive smoking cessation clinic follow-up group (intervention group, patients were informed on the importance and methods to quit smoking at the first visit and reminded for that at months interval for 6 months, n = 70) and the conventional treatment group (control group, n = 70). After 6 months, the smoking status, cardiovascular event rates, drug usage, out-patient medical costs and quality of life were compared between the two groups.
RESULTSAge, gender, concomitant diseases, drug usage were similar between the two groups at baseline (all P > 0.05). After 6 months, smoking quit rate [34.2% (24/70) vs. 5.7% (4/70), P < 0.01], drug use rates: lipid-lowering drugs [95.3% (67/70) vs. 80.4% (56/70)], β blockers [82.4% (57/70) vs. 41.3% (28/70)], and ACEI/ARB [61.4% (43/70) vs. 34.4% (24/70)] were significantly higher in the intervention group than in the control group, while total cardiovascular event rates [21.4% (15/70) vs. 47.1% (33/70), P < 0.01] and out-patient medical costs (3789.3 RMB vs. 4984.2 RMB, P < 0.01) were significantly lower in the intervention group than in the control group. The quality of life scores derived from MYO health survey questionnaire was significantly higher in the intervention group than in the control group (P < 0.01). The top three reasons responsible for continuous smoking for all patients failed to quit smoking were: (1) others smoked more than me and still alive and healthy [90.3% (56/62)]; (2) smoking helped me to keep relaxed and reduce trouble in daily work and life [70.9% (44/62)]; (3) smoking was essential while chatting and drinking with friends [66.1% (41/62)]. The overall satisfactory rate to this smoking cessation program was 42.8% and the satisfactory rate was up to 50.0% by patients.
CONCLUSIONSIntensive outpatient smoking cessation follow-up program can significantly improve the smoking cessation rates, the guideline drug use rate and the quality of life while reduce medical costs for coronary heart disease patients.
Aged ; Aged, 80 and over ; Coronary Disease ; Female ; Humans ; Male ; Middle Aged ; Outpatients ; Smoking Cessation ; methods
10.Mechanism of catgut embedding at back- points for nonalcoholic steatohepatitis based on IKK/IKB/NF-κB signaling pathway.
Xiao-Ling ZHOU ; Nong TANG ; Teng WU ; Yue ZHANG ; Qiao CHEN ; Wan-Hua WEI ; Ya-Ni ZHOU ; Dan-Xuan ZHANG ; Feng-Chuan QIN ; Wei YANG
Chinese Acupuncture & Moxibustion 2020;40(1):59-66
OBJECTIVE:
To explore the mechanism of catgut embedding at back- points on nonalcoholic steatohepatitis (NASH) in rats based on IKK/IKB/NF-κB signaling pathway and downstream inflammatory factors.
METHODS:
Eighty SPF SD rats were selected, among them 10 rats were selected divided into a normal group (group A), and the remaining 70 rats were fed with high-fat diet to establish NASH model. At the end of 12 weeks, 10 rats were randomly selected to verify whether the model establishment was successful. Then the remaining 60 rats were randomly divided into a model group (group B), a catgut embedding at back- points group (group C), a catgut embedding at abdominal points group (group D), an acupuncture at back- points group (group E), a sham catgut embedding group (group F) and a western medication group (group G), 10 rats in each group. The rats in the group C were treated with catgut embedding at "Ganshu" (BL 18), "Pishu" (BL 20), "Weishu" (BL 21) and "Shenshu" (BL 23); the rats in the group D were treated with catgut embedding at "Daheng" (SP 15), "Fujie" (SP 14), "Huaroumen" (ST 24) and "Tianshu" (ST 25); the rats in the group E were treated with acupuncture at the same acupoints as the group C; the rats in the group F were treated with catgut embedding at back- points but the needle did not enter subcutaneous tissue gamma; the rats in the group G were treated with intragastric administration of vitamin E capsule. All the treatment was given for 4 weeks. The rats in the group A were fed with normal diet until the end of 16 weeks without any intervention. The rats in the group B continued to be fed with high-fat diet until the end of 16 weeks. After the intervention, the liver index was calculated; the liver histomorphology was observed by HE staining; the liver function [alanine aminotransferase (ALT), gamma glutamyl transferase (γ-GGT), alkaline phosphatase (ALP)] and blood lipid [serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL)] were measured by serum biochemistry. The serum levels of TNF-α, IL-6 and IL-1βwere detected by ELISA, and the expressions of IKK-α, NF-κBp65, IL-6, IL-1β and TNF-α proteins in liver tissue were detected by Western blot. The temperature of the conception vessel and the governor vessel was measured by infrared thermography.
RESULTS:
Compared with the group A, the obvious steatosis and inflammatory cell infiltration were observed in the group B, and the body weight, liver wet-weight and liver index were all increased (<0.01). Compared with the group B, the liver tissue morphology in the group C, the group D, the group E and the group G was improved in varying degrees, and the liver index was decreased (<0.05), which was the most significant in the group C (<0.05). Compared with the group A, the ALT, γ-GGT, ALP, TG, TC, LDL, TNF-α, IL-6 and IL-1β were all increased in the group B (<0.01); compared with the group B, the ALT, γ-GGT, ALP, TG, TC, LDL, TNF-α, IL-6 and IL-1β in all intervention groups were all decreased in varying degrees (<0.01, <0.05), which was the most significant in the group C (<0.01). Compare with the group A, the expressions of IKK-α, NF-κBp65, TNF-α, IL-6 and IL-1βproteins in the group B were all increased (<0.01); compared with the group B, the expressions of IKK-α, NF-κBp65, TNF-α, IL-6 and IL-1βproteins in all intervention groups were decreased in varying degrees (<0.05), which was the most significant in the group C (<0.01). Compared with the group A, the temperature of the conception vessel and governor vessel was decreased in the group B (<0.01). Compared with the group B, the temperature of the conception vessel and governor vessel was all increased in the group C, the group D and the group E (<0.01); the temperature of the conception vessel in the group C was similar to that in the group D (>0.05), while the temperature of the governor vessel in the group C was superior to that in the group D (<0.05).
CONCLUSION
The catgut embedding at back- points might inhibit the activation of IKK/IKB/NF-κB signaling pathway to interrupt the inflammatory cascade, and reduce the "second hit" of inflammatory factors on liver, which could slow down NASH progress and prevent and treat NASH.
Acupuncture Points
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Animals
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Catgut
;
NF-kappa B
;
Non-alcoholic Fatty Liver Disease
;
Rats
;
Rats, Sprague-Dawley
;
Signal Transduction