1.The study of using siRNA technology to silence expression of LT gene of Enterotoxigenic Escherichia coli
Rui FU ; Hua LIU ; Yong DUAN ; Yuming WANG ; Bin SHAN
International Journal of Laboratory Medicine 2015;(8):1009-1011,1014
The distinctive LT siRNAs were designed according to the LT sequence .During the process of cultivation ,siRNA targeting the LT gene ,non‐specific control siRNA ,negative control siRNA and culture medium were added into siRNA group (siRNA‐LT1 group , siRNA‐LT2 group) ,siRNA‐coa3 group ,siRNA‐NC group and blank control group ,respectively ,and three times in each group (1 nmol each time) .After siRNA added at the first time ,bacteria was collected in 45 min (A) ,90 min (B) and 135 min (C) time points .The expression of mRNA in three time points (A ,B and C) were detected by real‐time fluorescence quantitative PCR .The protein level of LT in siRNA‐LT1 group ,siRNA‐LT2 group and blank control group were detected by Western blot in three time points .Results The results of real‐time fluorescence quantitative PCR showed that inhibition of siRNA‐LT1 on the expression of LT mRNA at the three time points(A ,B and C)were 70 .9% ,70 .1% ,72 .5% respectively ,and inhibition of siRNA‐LT2 on the ex‐pression of LT mRNA at the three time points(A ,B and C)were 70 .1% ,69 .2% and 70 .5% respectively .In the three time points (A ,B and C)the inhibition rate of the expression of LT mRNA in siRNA‐LT1 group and siRNA‐LT2 group were statistically lower than that in the siRNA‐NC group ,siRNA‐coa3 group and blank control group (P<0 .05) .The results of Western blot showed that in siRNA‐LT1 group the inhibitory rate of expression of LT protein in the three time points were 43 .1% ,18 .4% and 5 .0% ,re‐spectively ;in the siRNA‐LT2 group were 38 .2% ,15 .4% and 30 .1% ,respectively .Conclusion The specific siRNA could inhibit the expression of LT gene in vitro .
2.Protective effects of histone deacetylase inhibitor on stress-induced myocardial injury in rats.
Lei WU ; Xiao-hua LIU ; Tian-hui WANG ; Rui-feng DUAN ; Xue-si ZHOU ; Hong-tao LIU ; Zhi-qing ZHANG
Chinese Journal of Applied Physiology 2015;31(3):193-196
OBJECTIVETo observe the protective effects of histone deacetylase inhibitor on stress-induced myocardial injury.
METHODSHealthy male Wistar rats were randomly divided into 3 groups( n = 6), and the stress-induced myocardial injury model was established with chronic restraint stress method. The protective effects of histone deacetylase inhibitor on stress-induced myocardial injury were observed with Trichostatin A (TSA) intervention. Histone acetylation levels in myocardium of rats were detected by Western blot method, spectrophotometry method was used to dynamically determine the activity of rat serum lactate dehydrogenase (LDH), serum creatine kinase isoenzyme-MB (CK-MB) and Caspase 3, and nagar Olsen staining were used to observe the early myocardial damage.
RESULTSRestraint stress could significantly reduce the level of histone acetylation of myocardium in rats, and TSA intervention could inhibit the stress-induced reduction of myocardial levels of histone acetylation. Restraint stress could cause the significant increase of serum LDH activity ( P < 0.05), serum CK-MB activity ( P < 0.05), and the Caspase 3 activity of myocardial tissue (P < 0.05), and early myocardial damage also occurred during restraint stress. ISA intervention could significantly reduce the serum LDH activity (P < 0.05), the serum CK-MB activity (P < 0.05), the activity of myocardial tissue caspase 3 induced by restraint stress (P < 0.05), and the stress-induced myocardial injury was also attenuated by TSA intervention.
CONCLUSIONThe histone deacetylase inhibitor TSA can protect stress-induced myocardial injury.
Acetylation ; Animals ; Cardiotonic Agents ; pharmacology ; Caspase 3 ; blood ; Creatine Kinase, MB Form ; blood ; Histone Deacetylase Inhibitors ; pharmacology ; Hydroxamic Acids ; pharmacology ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Restraint, Physical ; Stress, Physiological
3.Meroterpenoids and isoberkedienolactone from endophytic fungus Penicillium sp. associated with Dysosma versipellis.
Jun-Wei LI ; Rui-Gang DUAN ; Jian-Hua ZOU ; Ri-Dao CHEN ; Xiao-Guang CHEN ; Jun-Gui DAI
Acta Pharmaceutica Sinica 2014;49(6):913-920
Seven meroterpenoids and five small-molecular precursors were isolated from Penicillium sp., an endophytic fungus from Dysosma versipellis. The structures of new compounds, 11beta-acetoxyisoaustinone (1) and isoberkedienolactone (2) were elucidated based on analysis of the spectral data, and the absolute configuration of 2 was established by TDDFT ECD calculation with satisfactory match to its experimental ECD data. Meroterpenoids originated tetraketide and pentaketide precursors, resepectively, were found to be simultaneously produced in specific fungus of Penicillium species. These compounds showed weak cytotoxicity in vitro against HCT-116, HepG2, BGC-823, NCI-H1650, and A2780 cell lines with IC 50 > 10 micromol x L(-1).
Berberidaceae
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microbiology
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Cell Line
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Cell Line, Tumor
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Humans
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Lactones
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isolation & purification
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pharmacology
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Monoterpenes
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isolation & purification
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pharmacology
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Penicillium
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chemistry
4.Three dimensional fetal heart structure study in fetal heart by sono-automatic volume count with spatio-temporal image correlation
Xiaohang ZHANG ; Rui LI ; Lingmin DUAN ; Yafang XUE ; Xuemei ZHANG ; Jianjun LIU ; Yan WANG ; Yanli GUO ; Hua ZHONG
Chinese Journal of Ultrasonography 2011;20(1):41-43
Objective To evaluate three dimensional fetal heart structure by sono-automatic volume count (SonoAVC) with spatio-temporal image correlation(STIC). Methods The heart volumes datas were acquired by STIC in 32 fetuses with postnatally confirmed diagnoses(20 cases of normal heart, 12 cases of complex congenital heart disease between 20 - 37 gestional weeks), then the volume datas were analyzed offline. SonoAVC software automaticly searched hypoechoic and anechoic structures, and assigned individual colors,and this can be corrected by manual splitting and/or removing and merging of individual segments.Results Individual segments of fetal heat could be separated,and digital casts were generated. The digital casts were obtained successfully in 4cases of transposition of the great arteries(TGA) ,2 cases of tetralogy of Follot,1 case of Ebstein's anomaly, 1 case of double-outlet right ventricle and 20 cases of normal heart.Conclusions Combination of STIC and SonoAVC can demonstrate the size, shape and connection of fetal cardiac cavities and great arteries in three dimensional spatial context. It has the potential both to help in obtaining strctural diagnostic,and to generate 3D visual displays for consultation and teaching.
5.Genetic polymorphisms of six Y chromosome short tandem repeat loci in Chinese Korean ethnic group.
Yong-ji ZHANG ; Song-hua LI ; Hong-rui DUAN
Chinese Journal of Medical Genetics 2012;29(1):82-86
OBJECTIVETo determine the allelic sequences and genetic polymorphism of six short tandem repeats (STRs) loci on Y chromosome, including DYS441, DYS442, DYS443, DYS444, DYS445 and DYS446 in Chinese Korean ethnic males from Yanbian region of Jilin province, China, and to construct a preliminary database.
METHODSAllele frequencies of the six STR loci in 205 Chinese Korean ethnic individuals were analyzed by multiplex polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE).
RESULTSRespectively, 8, 7, 7, 5, 6 and 9 alleles were detected for each of the locus. Together they have formed 151 haplotypes, with a diversity of 0.9937.
CONCLUSIONThe six STR loci included in this study were found to be highly polymorphic, and may provide useful markers for genetic analysis.
Alleles ; China ; Chromosomes, Human, Y ; Ethnic Groups ; genetics ; Gene Frequency ; Humans ; Korea ; ethnology ; Male ; Polymorphism, Genetic ; Tandem Repeat Sequences
6.Changes in learning and memory functions in rats with status epilepticus and generalized nonconvulsive status epilepticus.
Wei-ping WANG ; Yan LOU ; Pan LI ; Rui-sheng DUAN ; Wei-hua CHEN
Journal of Southern Medical University 2008;28(2):255-259
OBJECTIVETo observe the changes in the learning and memory functions and the hippocampal expression of phosphorylated cAMP response element-binding protein (pCREB) in rats with status epilepticus and generalized nonconvulsive status epilepticus.
METHODSStatus epilepticus (SE) and generalized nonconvulsive status epilepticus (GNCSE) was induced by pentylenetetrazol kindling in SD rats, and the learning and memory function changes of the kindled rats were assessed by means of Morris water-maze test and Y-maze test with alternative electric stimulation. Immunocytochemistry was used for analysis pCREB protein expression in the hippocampus of the rats.
RESULTSIn Morris water-maze test, the rats with SE showed prolonged mean escape latency (P<0.05), shortened swimming time in the platform quadrant (P<0.05), and reduced number of times of platform crossing (P<0.05) in the short term after kindling. But these changes were reversed and became normal a month after the kindling (P>0.05). In the Y-maze test with alternative electric stimulation, the total error (TE) of SE rats increased significantly in the short term after epilepsy (P<0.05), but recovered the normal level a month after kindling (P>0.05). The GNCSE rats showed prolonged mean escape latency at only certain time periods (P<0.05) in the short term, but with swimming time in the platform quadrant and number of platform crossings similar to the control group (P>0.05). The short-term TE of GNCSE rats increased significantly (P<0.05), but in the long term, TE was similar to that in the control group (P>0.05). The expression of pCREB decreased significantly in SE group in comparison with the control group in the short term.
CONCLUSIONEpileptic seizures can lead to learning and memory function impairment in rats, and SE seems to cause greater impact than GNCSE on the learning and memory functions. pCREB might be involved in the pathophysiology of learning and memory deficit in epileptic rats.
Animals ; CREB-Binding Protein ; metabolism ; Hippocampus ; metabolism ; physiopathology ; Kindling, Neurologic ; Maze Learning ; Memory Disorders ; physiopathology ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; metabolism ; physiopathology
7.Summary and analysis of safety warning on clinical application of anti-cold Chinese patent medicine preparations.
Xiao-xiao LAI ; Hua LIN ; Yi-ni LUO ; Ying-yan WANG ; Xiao-hong DUAN ; Lin WANG ; Rui LUO ; Yan-hong CHEN
China Journal of Chinese Materia Medica 2015;40(8):1594-1600
In China, many surveys have shown that most people do not have a correct understanding about cold and administration of anti-cold Chinese patent medicine preparations. The author conducted a systematic summary and analysis on the actual application of anti-cold Chinese patent medicine preparations as well as the warning on safe application of anti-cold Chinese patent medicine preparations in Clinical Medication Information of China Pharmacopoeia, in the expectation of reducing the blind application of anti-cold Chinese patent medicine preparations and providing traditional Chinese medicine pharmacists new ideas in monitoring the safe application of exterior syndrome-relieving Chinese patent medicine preparations.
China
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Common Cold
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drug therapy
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Drugs, Chinese Herbal
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adverse effects
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chemistry
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therapeutic use
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Humans
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Nonprescription Drugs
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adverse effects
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chemistry
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therapeutic use
8.Analysis of epidemiologic feature and genetic sequence of Sapovirus in China.
Zhao-Rui CHANG ; Miao JIN ; Na LIU ; Hua-Ping XIE ; Shu-Xian CUI ; Qing ZHANG ; Zhao-Jun DUAN
Chinese Journal of Virology 2009;25(2):113-116
To investigate epidemiologic feature and genetic variance of Sapovirus among children in China, fecal specimens were collected from children under 5 years old with acute diarrhea from Feb 2006 to Jan 2007 in nine provinces including Anhui, Fujian et al. A total of 1,110 fecal samples were detected for Sapovirus by reverse transcriptase-PCR (RT-PCR). Ten samples (0.9%) were positive for Sapovirus. The PCR products were then sequenced and analysed by phylogenetic tree. The results indicated that the detected Sapovirus strains were classified into two genogroups and three genotypes, including G I/1, G I/3, G II/3.
Astroviridae Infections
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epidemiology
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etiology
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genetics
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Base Sequence
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Caliciviridae Infections
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epidemiology
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China
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epidemiology
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Diarrhea
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classification
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virology
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Feces
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virology
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Gastroenteritis
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epidemiology
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etiology
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virology
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Genetic Variation
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Humans
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Molecular Sequence Data
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Phylogeny
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Reverse Transcriptase Polymerase Chain Reaction
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Sapovirus
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classification
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genetics
9.Association of Ureaplasma urealyticum colonization with development of bronchopulmonary dysplasia: a systemic review and meta-analysis.
Xiao-dan, ZHENG ; Dan, LI ; De-hua, YANG ; Xuan, XIANG ; Hong, MEI ; Jia-rui, PU ; Qiang-song, TONG ; Li-duan, ZHENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):265-9
There is controversy regarding the roles of Ureaplasma urealyticum (U. urealyticum) colonization in the development of bronchopulmonary dysplasia (BPD). This study explored the association between U. urealyticum and bronchopulmonary dysplasia at 36 weeks post-menstrual age (BPD36). Studies published before December 31, 2013 were searched from Medline, Embase, Ovid, Web of Science, and Cochrane databases, with the terms "Ureaplasma urealyticum", "chronic lung disease", or "BPD36" used, and English language as a limit. The association between U. urealyticum colonization and BPD36 was analyzed with RevMan 4.2.10 software, using the odds ratio (OR) and relative risk (RR) for dichotomous variables. Out of the enrolled 81 studies, 11 investigated the BPD36 in total 1193 infants. Pooled studies showed no association between U. urealyticum colonization and subsequent development of BPD36, with the OR and RR being 1.03 (95% CI=0.78-1.37; P=0.84) and 1.01 (95% CI= 0.88-1.16, P=0.84), respectively. These findings indicated no association between U. urealyticum colonization and the development of BPD36.
10.Efficacy of infrared laser combined with alpha zinc sulfate in the treatment of diabetic peripheral neuropathy
Ai-Ling YI ; Rui-Hua DUAN ; Shu-Fang GAN ; Wei CHEN ; Guang DU
The Chinese Journal of Clinical Pharmacology 2015;(3):166-168
Objective To observe the clinical efficacy of infrared laser combined with alpha-zinc sulfate for the treatment of diabetic peripheral neuropathy ( DNP).Methods A total of 106 patients with DPN were randomly divided into observation group ( n =53 ) and control group (n=53).Besides base hypoglycemic drug therapy, patients in the control group received alpha-zinc sulfate injection 0.6 g once a day.The patients in observation group were given the above treatment plus anodyne therapeutic apparatus treatment.The treatment period lasted for 12 days.The data of clinical symptoms, temperature, and nerve conduc-tion velocity of two groups were observed before and after treatment.Results The effective rate and total effective rate were 52.83% and 90.57%in the observation group, significantly better than 41.51%and 86.66%in control group ( P<0.05 ).The data of peroneal nerve and sural nerve conduction velocity in observation group were much signifi-cantly improved than before treatment(P <0.05).No significant adverse effect was observed.Conclusion Infrared laser combined with alpha-zinc sulfate is effective on the treatment of diabetic peripheral neuropathy.