2.Preparation of 99Tcm labeled survivin mRNA antisense PNA and gene imaging in nude mice bearing lung carcinoma A549 xenografts
Xin-ming, ZHAO ; Meng, DAI ; Ya-li, LIU ; Jian-fang, WANG ; Jing-mian, ZHANG ; Ying-chen, WANG ; Zhao-qi, ZHANG ; Chun-nuan, DAI ; De-zhi, LI
Chinese Journal of Nuclear Medicine 2011;31(5):339-343
Objective To prepare the 99Tcm-survivin mRNA antisense peptide nucleic acid (PNA)and investigate its value as a gene imaging agent in tumor bearing mice and early diagnosis in tumor.Methods Survivin mRNA antisense PNA and mismatch PNA were synthesized.Four amino acids (Gly- (D)Ala-Gly-Gly) and Aba (4-aminobutyric acid) were linked to the 5' end of PNA.Gly- (D)Ala-Gly-Gly served as a chelating moiety for strong chelation of 99Tcm and Aba acted as a spacer to minimize the steric hindrance.PNAs were labeled with 99Tcm by the ligand-exchange method.The labeling efficiency and radiochemical purity were measured by HPLC and ITLC methods.There were five BALB/c nude mice bearing human lung carcinoma ( A549 ) in each of antisense PNA and mismatch PNA groups.Gene imaging of 99Tcm-survivin mRNA antisense and mismatch PNAs were performed at 1,2 and 4 h post the injection,respectively,and the T/NT ratio was measured by the method of ROI.The statistical comparisons of average values were performed with the two-group t-test for independent sample by SPSS 13.0.Results The product kept stable in vitro.The labeling efficiency of 99Tcm-survivin mRNA antisense PNA was (95.48 ±1.92)% and more than 85% after the incubation for24 h in serum.The radiochemical purity was > 95%.The labeling efficiency of mismatch PNA was similar to the antisense PNA.99Tcm-survivin mRNA antisense PNA was especially uptaken by tumor lesion,and its accumulation reached the top at 4 h post the injection.T/NT ratios at 1,2,and 4 h were 2.70 ± 0.28,3.44 ± 0.35,4.21 ± 0.63,respectively.In the comparison,the T/NT ratio of 99Tcm-survivin mRNA mismatch PNA at 4 h (3.12 ±0.50) was significantly lower (t =2.918,P =0.019).Conclusions 99Tcm-survivin mRNA antisense PNA has high labeling efficiency,good stability and no need of purification.Its characteristic of especial uptake by tumor lesion provides the potential value in early diagnosis of tumor.
3.Virological surveillance on hemorrhagic fever with renal syndrome in Hunan province in 2006.
De-Fang DAI ; Hong ZHANG ; Yun-Zhi LIU ; Yi-Wei HUANG ; Li-Dong GAO ; Fu-Qiang LIU ; Ge ZENG
Chinese Journal of Epidemiology 2007;28(12):1194-1197
OBJECTIVETo understand the rate of viral carrying status among rodents as well as genotypes and distribution of Hantaviruses (HV) isolated in Hunan province.
METHODSWith DFA, the HV antigen in lung tissues of rodents was detected. The total viral RNA was extracted from the lung tissues of the HV infected rats and amplified with reverse transcrition-polymerase chain reaction (RT-PCR), using the HV genotype specific primers. The amplified genes were then sequenced and subjected to genotyping and homologic analysis.
RESULTSThe average density of rodents was 3.15% and the virus carrying rate among rodents was 1.31%. Data from genotype analysis showed that the HV isolated from seven lung specimens taken from Rattus norvgicus, Apodemus agraius, Mus musculus, Rattus flavipectus among indoor rodents in Shaodong and Liuyang belonged to HV type II (SEOV), and one isolated from Apodemus agraius in Shaungfen belonged to HV type I (HTNV) among outdoor rodents. Six strains were sequenced successfully and the homology between six srains was 88.3%-100%. The homology of HN1, HN2, HN4, HN6 came from Liuyang and the HN7 and HN8 from Shaodong were both 100% while the homology between L99 and the strains from Liuyang and Shaodong were 94.4% and 88.3% respectively.
CONCLUSIONHV type II (SEOV) and the HV type I (HTNV) were all existed in Hunan province while SEOV was the main genotype.
Animals ; Genotype ; Hantavirus ; classification ; genetics ; Hemorrhagic Fever with Renal Syndrome ; virology ; Phylogeny ; RNA, Viral ; genetics ; Rats ; Reverse Transcriptase Polymerase Chain Reaction
4.Clinical observation on electroacupuncture combined with catgut implantation at acupoints for treatment of simple obesity of heart and spleen deficiency type.
Chun-lin TANG ; De-chun DAI ; Gui-feng ZHAO ; Wei-fang ZHU ; Lin-feng MEI
Chinese Acupuncture & Moxibustion 2009;29(9):703-707
OBJECTIVETo observe the interventional effect of electroacupuncture combined with catgut implantation at acupoints for treatment of simple obesity of heart and spleen deficiency type.
METHODSSixty five cases were randomly divided into an observation group (33 cases) and a control group (32 cases). The observation group was treated with electroacupuncture combined with catgut implantation at acupoint therapy, the electroacupuncture was applied at Zhongwan (CV 12), Xiawan (CV 10), Guanyuan (CV 4), Tianshu (ST 25), ect. and catgut implantation was given at Zhongwan (CV 12), Tianshu (ST 25), Qihai (CV 6), etc. The control group was treated with electroacupuncture only. The body weight, body mass index (BMI), waistline, waist hip ratio (WHR), Pittsburgh sleep quality index (PSQI), Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD, 17 items) were evaluated before and after treatment, and these were also compared with those of 35 nomal cases.
RESULTSThe total effective rate of 93.9% in the observation group was higher than that of 84.4% in the control group (P < 0.05); the body weight, BMI, waistline, WHR, PSQI, HAMD and HAMA of simple obesity cases were obviously higher than those of normal cases (all P < 0.05). The scores of above indexes were all obviously decreased in both groups after treatment (all P < 0.05), and the improvement was more significant in observation group (P < 0.05).
CONCLUSIONThe sleep quality reduction and mental and psychology disorder exist in simple obesity patients, and electroacupuncture combined with catgut implantation at acupoints can reduce weight effectively, and at the same time improve the sleep quality and regulate psychological state.
Acupuncture Points ; Adult ; Catgut ; Electroacupuncture ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Obesity ; physiopathology ; psychology ; surgery ; therapy ; Prostheses and Implants ; Prosthesis Implantation ; Spleen ; physiopathology ; Treatment Outcome ; Young Adult
5.Detoxification mechanism of promotion of drug clearance by activation of pregnane X receptor in situation of drug overdose
Li-Bo DAI ; Ping-Fei FANG ; Huan-De LI
The Chinese Journal of Clinical Pharmacology 2018;34(4):454-457
Objective To investigate the effects of regular dose and toxic dose of quetiapine (QTP) on PXR signaling and downstream CYP3A4 and P-gp mRNA expression in rat brain and liver and also further investigate the effects of PXR agonist (dexamethasone,DEX) on the mRNA expression of PXR,CYP3A4 and P-gp in these tissues in context of drug overdose to illustrate the potential detoxication mechanisms of PXR activation in the brain and liver.Methods Normal group was intraperitoneally injected with QTP 10 mg · kg-1 once,intoxication modeling group was intraperitoneally injected with QTP 100 mg · kg-1 once,and the PXR activation experimental group was intraperitoneally injected with DEX 2.5 mg · kg-1 · d-1 for consecutive 4 days.SD rats were randomly divided into 4 groups:blank group,normal group,model group,and experimental group.SD rats were dosed daily by the vehicle or dexamethasone 2.5 mg · kg-1 · d-1 for 4 days,and were intraperitoneally injected with the vehicle or QTP 10 or 100 mg · kg-1 on the fourth day.After the 4 days of drug treatment,the rats were sacrificed 12,24,48 h after the last dose.The mRNA expression of PXR,cytochrome P4503A4(CYP3A4) and P-glycoprotein (P-gp) of rats in 4 groups were assessed by Real-time PCR method.Results After administration QTP for 24 h,the expression of PXR mRNA in liver in blank group,normal group,model group,and experimental group were (15.8 ± 0.8) × 10-3,(27.8 ± 2.4) × 10-3,(33.3 ± 1.4) × 10-3,(49.2 ± 2.0) × 10-3,the difference between blank group and normal group was statistically significant (P < 0.05),the difference between normal group and model group was statistically significant (P < 0.05),and the difference between model group and experimental group was statistically significant (P < 0.05).The factors in CYP3A4 and P-gp in liver of rats were same with PXR in liver,and the factors in prefrontal cortex and hippocampus of rats were same with in liver.Our data showed that QTP dose-dependently increased the expression of PXR,CYP3A4 and P-gp in both brain and liver.In addition,we found that the combination use of DEX along with the toxic dose of QTP accelerated the drug-induced mRNA expression of PXR,CYP3A4 and P-gp.Conclusion QTP itself could induce the signaling pathway of PXR-CYP3A4/P-gp,which was accelerated by the adjunctive use of PXR agonist,indicating that activation of PXR signaling pathway could activate detoxification system quickly and effectively,resulting in quick detoxification of xenobiotic and protection of organs.
6.Reduced estimated glomerular filtration rate and proteinuria are associated with increased cardiovascular events rate in octogenarian population
Hua WANG ; Dong-Fang DAI ; Xue-Zhai ZENG ; Jie-Fu YANG ; De-Ping LIU ; Ji-Hong YANG
Chinese Journal of Cardiology 2013;41(10):845-849
Objective To determine the relationship between estimated glomerular filtration rate (eGFR) and proteinuria with cardiovascular events in subjects aged 80 years or older.Methods Data for this retrospective prognostic study were drawn from the patient database for routine checkup in Beijing hospital between January 2001 to December 2001.Baseline eGFR and proteinuria were evaluated in 340 subjects [mean age:(85.6 ± 4.0) years].eGFR was calculated using the modified abbreviated MDRD equations based on the Chinese chronic kidney disease patients.The subjects were divided into normal renal function group and reduced renal function group (eGFR < 60 ml · min-1 · 1.73 m-2).The subjects were divided into subjects without proteinuria and subjects with proteinuria group.Cardiovascular events included cardiovascular death,nonfatal myocardial infarction,nonfatal stroke.Results The proportion of reduced renal function was 36.8% (125/340).The proportion of proteinuria was 10.3% (35/340).The proportion of reduced renal function or proteinuria was 41.8% (142/340).Follow-up time was 79 months (40-114 months).Cardiovascular events rate was significantly higher in reduced renal function group than in normal renal function group [37.6% (47/125)vs.26.2% (55/210),P <0.05] and in proteinuria group than in without proteinuria group [50.0% (17/34) vs.28.2% (85/301),P < 0.01].Cox multivariate analysis revealed that both eGFR (HR =0.978,95% CI:0.961-0.994,P < 0.05) and proteinuria (HR =2.049,95% CI:1.132-3.709,P < O.05) were independent risk factors for cardiovascular events after adjusting for age,gender,triglyceride,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,uric acid,hypertension,coronary heart disease,diabetes mellitus.Conclusions Reduced eGFR and presence of proteinuria are independent risk factors for cardiovascular event in subjects aged 80 years or older.eGFR and proteinuria can thus be used for cardiovascular event risk stratification in subjects aged 80 years or older.
7.A review of operative treatment of hypospadias in twelve years.
De-Xin DONG ; Song CEN ; Yuan-Xiao LIU ; Jiang ZHONG ; Wei-Fu WANG ; Xiong FENG ; Bu-Min DAI ; An-Fang WANG ; De-Hai CAI ; Wei HUANG ; Jian CHEN ; Yang WANG
National Journal of Andrology 2002;8(6):422-424
OBJECTIVESTo evaluate the relationship between the operative methods and the therapeutic results of the patients with hypospadias.
METHODSNine operative types and different tissue materials taken in hypospadias operations in the past 12 years were retrospectively analyzed.
RESULTSThe operative types and tissue materials had significantly effects on the therapeutic results besides the clinical experience of the operators. The cure rates of Tunneltron Urethroplasty, Preputial island flap urethroplasty and Bladder mucosa graft urethroplasty were 86.4%, 83.3% and 83.0% respectively.
CONCLUSIONSAll the nine types and different tissue materials of Hypospadias operations have its own advantages and disadvantages. They are worth further study and improvement.
Adolescent ; Adult ; Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Infant ; Male ; Mucous Membrane ; transplantation ; Penis ; surgery ; Surgical Flaps ; Treatment Outcome ; Urethral Stricture ; surgery ; Urinary Bladder ; transplantation ; Urinary Fistula ; surgery ; Urologic Surgical Procedures, Male ; methods
8.Comparative study on imaging of 99 Tcm-survivin mRNA antisense peptide nucleic acid in tumor and inflammation animal models
Xin-ming, ZHAO ; Ya-li, LIU ; Meng, DAI ; Xiu-chun, REN ; Jian-fang, WANG ; Jing-mian, ZHANG ; Ying-chen, WANG ; Zhao-qi, ZHANG ; Xiu-juan, ZHAO ; Chun-nuan, DAI ; De-zhi, LI
Chinese Journal of Nuclear Medicine 2011;31(6):364-367
ObjectiveTo investigate the value of 99Tcm labeled survivin mRNA antisense peptide nucleic acid (PNA) as an imaging agent in the specific diagnosis for carcinoma.MethodsSurvivin mRNA antisense PNA was labeled directly with 99Tcm by the ligand-exchange method.Twenty nude mice with lung carcinoma A549 xenografts were randomly divided into 4 groups.Three groups were used for biodistribution study and one group was used for imaging study.Other twenty mice infected by staphylococcus aureus underwent the same procedure.The biodistribution and imaging of 99Tcm-survivin mRNA antisense PNA was studied at 1,2 and 4 h respectively after the intravenous injection in nude mice bearing lung carcinoma A549 xenografts or inflammation models.SPSS 13.0 was used in the study and all data were analyzed by t test.ResultsBiodistribution results showed that the highest radioactivity was found in the liver,and then in the kidney.Four hours after the administration of the imaging agent,the radioactivity ratios of target-tonon target (T/NT,tumor or inflamumatory lesions to the contralateral regions) in tumor model group were significantly higher than those in inflammation model group ( 3.69 ± 1.13 vs 2.03 ± 0.47,t =3.01,P =0.02 ).Tumors were clearly visible in the tumor model groups at 0.5 h and still clearly seen at 4 h after the injection of antisense PNA.On the contrary,inflammatory lesions could not be seen clearly.Conclusion 99Tcm labeled survivin mRNA antisense PNA can be used to distinguish tumor from inflammation and it may provide a new feasible method for specific tumor diagnosis.
9.Expression of vasoactive intestinal peptide in peripheral blood of children with hand, foot and mouth disease.
Jin-Song REN ; Hao-Miao SUN ; Lei ZHANG ; Jing-De LIN ; Cheng WEN ; Dai-Hua FANG
Chinese Journal of Contemporary Pediatrics 2016;18(11):1106-1110
OBJECTIVETo investigate the expression of vasoactive intestinal peptide (VIP) in peripheral blood of children with hand, foot and mouth disease and its significance.
METHODSAccording to the condition of the disease, 86 children with hand, foot and mouth disease were classified into phase 1 group (19 children) and phase 2 group (67 children). ELISA was used to measure the concentrations of plasma VIP, interferon-γ (IFN-γ), and interleukin-4 (IL-4) in peripheral blood. Flow cytometry was used to measure CD3, CD4, and CD8T lymphocyte subsets. RT-PCR was used for qualitative detection of enterovirus 71 (EV71) RNA in stool.
RESULTSCompared with the phase 1 group, the phase 2 group had a significantly higher positive rate of EV71-RNA (P<0.05) and significantly higher serum levels of IgG, IgA, IgM, and C3 (P<0.05). The phase 2 group had significantly lower proportions of peripheral CD3, CD4, and CD8T lymphocyte subsets than the phase 1 group (P<0.05), as well as significantly lower proportion of peripheral B cells and CD4/CD8ratio than the phase 1 group (P<0.05). The phase 2 group also had a significantly lower concentration of VIP in peripheral blood than the phase 1 group (P<0.05). In the 86 children with hand, foot and mouth disease, the concentration of VIP in peripheral blood was positively correlated with the proportion of CD4T lymphocyte subset and CD4/CD8ratio (r=0.533 and 0.532 respectively; P<0.05).
CONCLUSIONSVIP may be an important marker of the severity of hand, foot and mouth disease.
Biomarkers ; CD4-CD8 Ratio ; Child, Preschool ; Female ; Hand, Foot and Mouth Disease ; immunology ; Humans ; Infant ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Severity of Illness Index ; Vasoactive Intestinal Peptide ; blood
10.An epidemiologle investigation of hantaviruses carried by rodent hosts in Hunan province.
Yong-zhen ZHANG ; Qi-you XIAO ; Ming-hui LI ; Yang ZOU ; Wei LV ; De-fang DAI ; Hua-xin CHEN
Chinese Journal of Epidemiology 2007;28(1):65-69
OBJECTIVETo explore the hantavirus infection and their genotype in rodents in Hunan.
METHODSHantavirus antigens in the rat lungs from Hunan province were detected by immunofluorescence assay. Partial S and M segment in antigen-positive samples were amplified by RT-PCR, and then sequenced. The phyologenetic trees were constructed for the analysis of genetic characters of hantavirus.
RESULTSA total of 344 rats were trapped in the main epidemic area of Hunan province, and hantavirus antigens were found in 6 of the 344 rats( 1.74% ).The phylogenetic trees constructed by partial S segment( nt 620-990) or partial G2 segment (nt 2001- 2301) showed that the hantaviruses carried by Rattus norvegicus, R . flabipectus and R. rattoides from Xiangxiang district were genetic subtype SEOV4. The virus carried by R. norvegicus in Ningyuan district was phylogenetically different from the known SEOV. The hantavirus carried by Mus musculus from Shimen district was genetic subtype HTNV4.
CONCLUSIONThe hantaviruses in the main epidemic areas in Hunan province mainly belonged to SEOV, and R. flabipectus and R. rattoides carried the same genotype of SEOV as R. norvegicus.
Animals ; China ; epidemiology ; Disease Reservoirs ; virology ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; epidemiology ; virology ; Mice ; Molecular Sequence Data ; Phylogeny ; Rats ; Rodentia ; virology