1.Construction of Subtractive cDNA Library of K562 Cells
Hanchun CHEN ; Qiao MENG ; Jiwei WANG
Journal of Chinese Physician 2001;0(01):-
Objective To construct the suppression subtractive library of hemin-inducing K562 cells for identifying the cDNA genes of globin synthesis regulatory factors expressed in K562 cells induced by hemin. Methods K562 cells were cultured under different concentration of hemin, both the percentage of positive benzidine staining cells and hemoglobin content were measured, the most reasonable concentration of hemin was chosen for inducing incubation of K562 cells. The poly (A) positive RNA (mRNA) was isolated from the hemin-induced K562 cells (tester) and non-induced K562 cells (driver) respectively, and double-strand cDNA molecules were synthesized by reverse transcription. After two times subtractive hybridization followed by two times polymerase chain reaction (PCR) amplification, the forward subtracted PCR products were ligated with pGEM T-Easy vector and the subtracted cDNA library was constructed. The library clones were selected by blue-white screening. The plasmid DNAs of the single positive colony were purified and digested by EcoRⅠ, and the inserts in plasmid were amplified by PCR. Results The maximum of positive benzidine staining cells percentage and hemoglobin content of K562 cells were obtained in 50?mol/L of hemin inducing condition. The suppression subtractive library of hemin-inducing K562 cells was constructed after K562 cells were induced by 50?mol/L of hemin. The library identification showed that the positive clones contain inserts in different length respectively. Conclusions The suppression subtracted cDNA library of hemin-inducing K562 cells were successfully constructed. This subtracted cDNA library combined with bioinformatics analysis can be used to explore structures and functions of hemin-inducing expression genes in K562 cells.
2.The scheme and application of educational reformation during pre-medical training of medical education for eight-year program
Zhangming WEI ; Xiaolei REN ; Jing LIU ; Yifan ZHU ; Chunxiang JIANG ; Hanchun CHEN ; Qingnan HE
Chinese Journal of Medical Education Research 2015;(8):780-782,783
Central South University began to implement the eight-year medical education in 2004, and has accumulated rich experience in the pre-medical education through continuous reform and improvement. Since 2012, Central South University has made a series of reformation, which is more conducive to the all-round development of medical students , on the pre-medical education . Through adding freshman courses, humanities courses, bilingual teaching courses, and applying aca-demic adviser institution, early scientific research training and open courses during the pre-medical education, Central South University has exercised and strengthened students' scientific research ability, the humanistic quality, English level, and made the pre-medical comprehensive examination for the shunt selection of students. Thus Central South University has improved the eight-year medical students' comprehensive quality, and provided an example of the reformation for medical education.
3.Identification of pathogenic microorganism by sequencing 16S rRNA gene.
Feizhou ZHU ; Liyu CHEN ; Hanchun CHEN
Journal of Central South University(Medical Sciences) 2013;38(10):1035-1041
OBJECTIVE:
To identify 14 bacteria by sequencing the 16S rRNA gene and establish the basis for clinical application in the future.
METHODS:
DNA samples of the 14 bacteria were extracted. The 16S rRNA genes were amplified by PCR and sequenced with common primers. The sequences of the 16S rRNA genes were aligned by online software Blastn in nucleotide database. The bacteria were identified according to the homology of their 16S rRNA genes.
RESULTS:
Twelve bacteria were classified to species, the other 2 bacteria were classified to genus.
CONCLUSION
16S rRNA gene sequence analysis is useful in identifying pathogenic bacteria.
Bacteria
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classification
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DNA Primers
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DNA, Bacterial
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genetics
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Polymerase Chain Reaction
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RNA, Ribosomal, 16S
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genetics
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Sequence Analysis, DNA
4.Endoscopic feasibility study and nasal septum median path of frontal sinus surgery.
Junwei MA ; Tingting LIU ; Wei LI ; Zhimin WANG ; Dongyi JIANG ; Hanchun CHEN ; Niankai ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2012;26(4):160-163
OBJECTIVE:
To discuss the feasibility of endoscopic frontal sinus surgery in the nasal septum median path.
METHOD:
(1) Sixty adult cadaveric heads fixed with formalin were CT scanned,and were three dimensional reconstruction. (2) Thirty adult cadaveric heads were sawn along the sagittal line close to the side of the nasal septum, then the important anatomic marks were observed and measured. (3) Combined with CT and anatomical data, thirty adult cadaveric heads were operated in different degree, and the damage of nasal septum and fila olfactoria were detected in the same time.
RESULT:
(1) The roots of middle nasal concha were simulated in the endoscopic frontal sinus surgery. The operation time, operative procedures, markers foundation, endoscopic back of posterior border of frontal sinus foundation and attached to the symphysis with cribriform plate and the top of ethmoidal sinus were recorded. (2) The intersection point formed by the level of middle nasal concha and the vertical of middle nasal concha corresponded with the nasal septum was called the M point. The distance from the M point to the horizon of the nasal bone was (20.07 +/- 6.21) mm, the distance from the M point to the first fila olfactoria was (24.38 +/- 7.68) mm, the distance from the first fila olfactoria to the posterior edge of frontal sinus was (9.57 +/- 2.73) mm, the distance from the root of the middle nasal concha to posterior edge of frontal sinus was (5.38 +/- 1.23) mm, the anteroposterior diameter of frontal sinus fundus was (7.62 +/- 2.45) mm, the transverse diameter of frontal sinus fundus was (9.41 +/- 3.37) mm, the seesaw diameter of frontal sinus partition was (16.97 +/- 3.23) mm, the anteroposterior diameter of frontal sinus partition was (12.34 +/- 2.23) mm. (3) The operation time through the nasal septum path was 105 minutes which combined with CT and anatomical measurements. 0 degrees endoscopy could be used to observe the frontal part of the lateral, posterior and top wall, while nasal septum remove should be finished with 30 degree endoscopy. The bottom of frontal sinus can be exposed and removed with 0 degree endoscopy. 3 cases of cadaveric frontal sinus lateral wall can not be observed with 70 degree endoscopy. 30 cases of cadaveric frontal sinus,some of the top and the lateral wall, anterior and posterior wall could be observed with 70 degree endoscopy, nasal septum damage range was about 2.23 cm x 2.59 cm, and no fila olfactoria damage was found.
CONCLUSION
Endoscopic frontal sinus surgery in the nasal septum median path is a good way to find frontal sinus.
Endoscopy
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methods
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Feasibility Studies
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Frontal Sinus
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diagnostic imaging
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surgery
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Humans
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Nasal Bone
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diagnostic imaging
;
surgery
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Nasal Septum
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diagnostic imaging
;
surgery
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Tomography, X-Ray Computed