2.Effect of RNA interference targeting for cyclin D1 gene on growth,cell cycle and apoptosis of K562
Yi-Yuan WAN ; Xiao-Long CAO ; Wen-Qi JIANG ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(07):-
Objective cyclin D1 gene plays a significant role in regulating cell cycle progression.Suppression of cyclin D1 protcin expression can effect on cellular proliferation,distribution of cell cycle and apoptosis.This study was to determine whether this effect also existed in chronic leukemia ceil line K562 by inhibiting the expression of cyclin D1 protein through RNA interference in vetro.Methods Plasmid vectors expressing small hairpin RNA (shRNA) targeting at cyclin D1 gene were constructed and transfected into K562 cells by chitosan,cyclin D1 protein was examined by using Western blot analysis.Inhibition of cellular proliferation was evaluated hy soft agar colony formation assay.The cell cycle and apoptosis were determined by flow cytometry.Results Expression of cyclin D1 protein was markedly down-regulated and capability of colony formation was suppressed after transfection with pshRNA-419 and pshRNA-575 at 48h.Down-regulation of cyclin D1 protein could effect on distribution of cell cycle arrested at G_0/G_1 phase and markedly induce apoptosis of K562 cells.But there had no above biological effects ob- served after transfection with blank vector and control vector of m-pshRNA-790.Conclusion Down-regulation of cyclin D1 expression can inhibit growth of K562 cells,and effect on distribution of cell cycle arrested at G_0/G_1 phase.The primary results suggest that cyclin D1 gene might serve as an effective target for the treatment of leukemia.
3.DNA aptamer selection in vitro for determining ketamine by FluMag-SELEX.
Mei-Qi SUN ; Fang-Qi CAO ; Xiao-Long HU ; Yu-Rong ZHANG ; Xin-Wei LU ; Li-Bo ZENG
Journal of Forensic Medicine 2014;30(5):346-349
OBJECTIVE:
To select specific DNA aptamer for determining ketamine by FluMag-SELEX.
METHODS:
Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity.
RESULTS:
Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine.
CONCLUSION
FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.
Aptamers, Nucleotide/metabolism*
;
DNA
;
DNA, Single-Stranded/genetics*
;
In Vitro Techniques
;
Ketamine/metabolism*
;
Oligonucleotides
;
SELEX Aptamer Technique/methods*
4.Preparation method of hyperoside from Flos Abelmoschus manihot
Shunli XIAO ; Min WANG ; Qi LI ; Qingyun CAO ; Rongkai GAO ; Honggui ZHANG
International Journal of Traditional Chinese Medicine 2017;39(8):733-736
Objective To study the preparation method and analytical technique of hyperoside from Flos Abelmoschus manihot.Methods Hyperoside was isolated and purified by solvent extract and chromatography, whose structure was determined by 1H-NMR and 13C-NMR. The purity was analyzed by TLC and HPLC.Results The TLC showed that the hyperoside had no impurity spot. The HPLC indicated that the purity reached more than 98.5%.Conclusions The mothod of isolation and purification for hyperoside reported in this paper was simple and economical.
5.Gas analysis of umbilical cord artery blood on predicating the prognosis of asphyxia neonate
Qi CAO ; Ge PAN ; Jinli MO ; Xiao HE ; Changliang NONG ; Fen HUANG
Clinical Medicine of China 2016;32(8):681-684
Objective To study the gas analysis of umbilical cord artery blood and radial artery blood on predicating the prognosis of asphyxia neonate?Methods From September 2014 to September 2015, 328 neonates were divided into groups by Apgar score:290 patients in the control group and 27 patients in the mild asphyxia group,11 patients in the severe asphyxia group?After birth,umbilical artery blood,radial artery blood gas analysis was perfomed, oxygenation index was calculated, Outcome of neonatal behavioral neurological assessment ( NBNA) in neonates with asphyxia was regular follow?uped,the relationship between pH value and umbilical artery blood gas analysis was analyzed?Results The pH, PO2, PCO2 and oxygenation index of umbilical cord blood and radial artery blood in the severe asphyxia group was(7?11±0?25,(73?93±23?35) mmHg,(51?36±16?37) mmHg,206?23±98?12),significant different than the mild group(7?24±0?05,(86?35 ±12?56) mmHg,(45?89± 9?21) mmHg,411?22±57?94) and the control group(7?28±0?08,(87?80±12?07) mmHg,(43?68± 6?45) mmHg,426?23±73?30)(P<0?05)?The pH,PO2,PCO2 and oxygenation index of umbilical cord blood and radial artery blood in the severe asphyxia group was(7?25±0?18,(74?66±24?09) mmHg,(51?42±17?83) mmHg,332?03±65?19),significant different than the mild group(7?31±0?09,(87?24 ±11?75) mmHg,(45?73±10?21) mmHg,405?67±82?65) and the control group(7?32±0?06,(87?99±11?81) mmHg,(42?84± 9?32) mmHg,439?89±60?76)(P<0?05)?The NBNA scores of the severe asphyxia group was (34?09±5?02) points,lower than the mild group(36?62±2?04)(F=21?65,P<0?05)?The NBNA scores showed significant relationship with umbilical cord blood pH in the severe asphyxia group( r=0?877,P<0?01)?Conclusion The pH,PO2 and oxygenation index of umbilical cord blood and radial artery blood was lower while PCO2 was markedly high in the severe asphyxia group than other groups?For neonates, there is a correlation between umbilical cord blood pH and NBNAs core, neonates borned with hypoxia and acidosis should monitor blood gas analysis and oxygenation index dynamically
6.High cumulative risk and clinical feature of intussusception in patients with Peutz-Jeghers syndrome
Qi TANG ; Ping ZHOU ; Xiao CHEN ; Xiaojian WANG ; Gaoping MAO ; Chuanping CAO
Chinese Journal of Digestion 2014;34(2):118-120
Objective To analyze the characteristics,risk and induced factors of intussusception in Peutz-Jeghers syndrome (PJS).Methods From March 2nd 2005 to May 25th 2013,a total of 130 patients with PJS were selected.The clinical data of patients were collected,which included gender,age,the diagnostic age of PJS,family history,the diagnostic age of intussusception,clinical features,location,treatment and the maximum diameter of polyps which caused intussusception.Kaplan-Meier was performed to analyze the cumulative risk of intussusception.Results Among 130 patients with PJS,90 cases had intussusception.The age when first time diagnosed was from four to 33,median age was 16.The cumulative risks of intussusception at 10,15,20,25 and 30 years were 12.308% (16/130),31.538%(41/130),51.538%(67/130),64.615%(84/130) and 66.923%(87/130).There was no significant difference in the cumulative risk of intussusception between male and female; with family history and no family history (both P>0.05).A total of 131 intussusception happened in 90 patients,of which diagnosed by surgery,imaging examination and history reviewer was 125,four and two times,respectively.The initial symptom of 111 times of intussusception was acute abdomen and 15 were abdominal pain and vomiting.The left five intussusception was found by regular:examination.One hundred and fifteen intussusception was in small intestine and 16 in colon.There was 127,three and one time treated with surgery,conservative treatment,endoscopic therapy (dual airbags intestinal endoscopic polypectomy),respectively.The maximum diameter of polyps which caused intussusception was from 15 to 70 mm,average 40 mm.Conclusions Intussusception of patients with PJS is at young age and with a high cumulative risk.Intussusception is generally caused by diameter over 15 mm polyps.
7.Radiography comparison of the pulmonary embolism
Zhan-Hong MA ; Ru-Ping DAI ; Cheng CAO ; Xiao-Ou QI ; Hua BAI ; Chen WANG ;
Chinese Journal of Radiology 1994;0(06):-
Objective To evaluate the application value of X-ray,echocardiogram,pulmonary perfusion scintigraphy,EBCT,Magnetic resonance Pulmonary angiography in diagnosis of PTE.Methods Twenty-five consecutive patients clinically diagnosed of having PTE were examined from july 2003 through March 2004. Patients underwent X-ray chest plain film, echoeardiogram, electronic beam computed tomographie (EBCT)angiography,ventilation-perfusion (V-P)seintigraphy,Magnetic resonance Pulmonary angiography (MRPA)and puhnonary angiography according to a strict diagnostic protocol.Two of the independent readers reviewed the pulmonary angiography and record all of the lobe and segmental involved in PTE and compared with other image method.Results Pulmonary angiography:all of the patients success underwent the technique,the pulmonary artery branch with PTE was in 556 of 775 branches (71.7%). Chest radiography had hints of diagnosis in 12 of 25 patients.Nine patients diagnosed with echocardiogram. Right heart enlargement was in 21,and pulmonary hypertension in 18.V-P scintigraphy revealed 247 segmental involved with PTE of 500 (52.0% ),and the sensitivity was 64.66% compare with the pulmonary angiography.There were 523 pulmonary branches involved PTE with EBCT pulmonary angiograpy of 775 branches,and the sensitivity was 94.06%.MRPA: 8 of 10 patients succeed in the technique, 155 branches of 248 were detected with PTE(62.5% ),the sensitivity was 81.29%.Conclusions EBCT is a high sensitivity method in diagnosis of PTE.Chest radiography and echocardiogram are the first-line modality of PTE.V-P scintigrapby is the valid compensation in diagnosis subsegmental pulmonary artery with PTE when EBCT miss diagnosis.Gd-CE-MRPA may be the second-line modality in diagnosis of PTE.
8.Analysis of point-selection pattern in acupuncture-moxibustion treatment of amblyopia
Zhao-Xia CAO ; An-Guo LIU ; Xing-Ke YAN ; Xiao-Juan LI ; Guo-Qi HUANG
Journal of Acupuncture and Tuina Science 2018;16(2):127-132
Objective:To analyze and summarize the characteristics and pattern in the selection of points and meridians by searching the clinical research literature about acupuncture-moxibustion treatment of amblyopia in the recent 17 years.Methods:By searching Chinese and English databases,such as China National Knowledge Infrastructure (CNKI),PubMed,etc.,the points in 52 articles in conformity with the requirements were analyzed by frequency statistics by the order of meridians,major points,and adjunct points to summarize the rules and characteristics of the point selection.Results:In 52 articles,there were 21 articles on ear acupuncture and 31 articles on acupuncture-moxibustion treatment.Ten meridians were involved in acupuncture-moxibustion treatment of amblyopia,and the leading 3 meridians were the Bladder,Stomach and Gallbladder Meridians.There were 34 points,10 extraordinary points and 37 ear points were used in acupuncture-moxibustion treatment of amblyopia.The top 10 major points were Jingming (BL 1),Taiyang (EX-HN 5),Cuanzhu (BL 2),Fengchi (GB 20),Baihui (GV 20),Hegu (LI 4),Guangming (GB 37),Sibai (ST 2),Chengqi (ST 1) and Sizhukong (TE 23).The top 5 adjunct points were Zusanli (ST 36),Shenshu (BL 23),Ganshu (BL 18),Sanyinjiao (SP 6) and Taixi (KI 3).Conclusion:Acupuncture-moxibustion treatment of amblyopia is characterized by the selection of the points mainly from yang meridians,based upon syndromes differentiation plus personal experience,and the points mainly around the eyes and by stressed use of ear points.
9.Influence of β-amyloid peptide on expressions of synaptophysin, dynamin Ⅰ and adaptor protein 180 in human neuroblastoma SH-SYSY cells
Ying CAO ; Yuan LIAO ; Yan XIAO ; Xiao-Lan QI ; Zhi-Zhong GUAN
Chinese Journal of Neuromedicine 2011;10(8):774-778
Objective To study the effect of β-amyloid peptide (Aβ1-42) on expressions of synaptophysin (Syn), dynamin Ⅰ (Dyn Ⅰ) and adaptor protein 180 (AP180) in human neuroblastoma SH-SY5Y cells. Methods Human SH-SY5Y neuroblastoma cells were treated with 0.01, 0.1, 1, 2 and 5 μmol/L Aβ1-42, and control cells were given no treatment. MTT [3-(4,5-dimethylthiazol- 2- yl) - 2, 5-diphenyltetrazolium bromide]reduction of the cells was measured by spectrophotometry. The protein levels of Syn, Dyn Ⅰ and AP180 in SH-SY5Y cells treated with 0.5 and 1 μmol/L Aβ1-42 were surveyed by Western blotting. The mRNA levels of Syn, Dyn Ⅰ and AP180 were detected by Real-time PCR in SH-SY5Y cells treated with 1 μmol/L Aβ1-42. Results SH-SY5Y cells showed obviously decreased reduction rates of MTT after exposure to Aβ1-42(0.1 μmol/L) as compared with the controls (P<0.05), and dose-dependent negative correlation was noted in these SH-SY5Y cells. The protein level of Dyn Ⅰ in cells treated with 0.5 μmol/L Aβ1-42 was significantly decreased as compared with that in controls (P<0.05). The protein and mRNA levels of Syn and Dyn Ⅰ in cells treated with 1 μ mol/L Aβ1-42 Were obviously decreased as compared with those in controls (P<0.05), but the levels of AP180 were not changed. Conclusion Aβ1-42 reduces the levels of Syn and Dyn Ⅰ in SH-SY5Y cells, which might be a mechanism in eonnection with cognitive deficit of AD.