1.Study on quality control of the next-generation sequencing data
Sheng LIU ; Zuren FENG ; Shan GAO
Military Medical Sciences 2014;(5):377-380
Objective To investigate the quality control of the next-generation sequencing data by analyzing some im-portant problems with quality control in order to provide high-quality data for the downstream data analysis .Method The software Fastq_clean was developed to clean the data and to statistically analyze the quality of the data.Result Using a Chrysanthemum transcriptome dataset , the function of the Fastq_clean software was illustrated .Conclusion Based on the Bioconducter package , the Fastq_clean software can accurately remove low quality nucleotides and Ns , adapter contamina-tion, possible rRNA and virus contamination .Moreover, it can keep the largest possible amount of high-quality data reads.
2.Retrovirus mediated IL-23 gene expression in mouse high-metastasis mammary cancer cells MA-891
Shuang LIU ; Yonglu FENG ; Baoen SHAN
Chinese Journal of Immunology 1985;0(03):-
Objective:The mouse mammary cancer cell line IL-23/MA-891 expressing IL-23 protein was set up and the effect of IL-23 on growth and other biological character of the cells was studied.Methods:The IL-23 gene was sequencely transfected into two packing cell lines (ecotropic ?2 and amphotropic PA317) by a retrovirus vector (LXSN),and the positive cellular clones were screened by G418.After infection to MA-891 cells with the culture supernatant of IL-23/PA317 cells and selected with G418,the IL-23/MA-891 cells to express IL-23 mRNA and protein was detected with RT-PCR,ELISA and immunocytochemical stairing,respectively.The expression of H-2Kb(MHCⅠ),I-Ab(MHC Ⅱ),CD80,CD86 and FAS was examined with flow cytometry.The ability to secrete IFN-? by mouse splenocytes stimulated with the culture supernatant of IL-23/MA-891 cells was detected with ELISA.The proliferation of MA-891,LXSN/MA-891 and IL-23/MA-891 cells was detected by MTT colorimetry and flow cytometry in vitro.Results:IL-23/MA-891 cells express IL-23 stably in mRNA and protein level.The expressing of H-2Kb(MHCⅠ),I-Ab( MHCⅡ),CD80,CD86 and FAS protein by flow cytometry were showing no difference in three kinds of cells.The proliferation rate of IL-23/MA-891 cells were showing little decresed,but statistically showing no difference with parental cells.The culture supernatant of IL-23/MA-891 cells induced secretion of IFN-? by mouse splenocytes were increased compared to parental cells.Conclusion:The cellular clone IL-23/MA-891 with high level expression of IL-23 was produced.
4.Effect of the imprinting loss of insulin like growth factor 2 gene during the differentiation from mouse embryonic stem cells to islet-like cells in vitro
Feng LIU ; Yuhuan PENG ; Jiazhen TANG ; Shan JANG
Chinese Journal of Tissue Engineering Research 2017;21(9):1362-1367
BACKGROUND: Insulin like growth factor 2 (IGF2) is an important embryonic mitosis growth promoting factor, whichplays a critical role in the process of maintaining normal cell growth. The gene expression of IGF2 is under epigeneticregulation in the way of genomic imprinting. Imprinting loss of IGF2 is likely to be associated with the abnormaldevelopment of the individual and tumorigenesis.OBJECTIVE: To investigate the effect of imprinting loss of IGF2 gene on the differentiation of mouse embryonic stemcells into islet-like cells.METHODS: Two kinds of mouse embryonic stem cells (SF1-G and SF1-1) were induced to differentiate into islet-likecells in vitro. The expression of Insulin gene was detected by real-time PCR and cell immunofluorescence. Theexpression of IGF2 was detected by the polymerase chain reaction-restriction fragment length polymorphism in the cellsbefore and after induced differentiation. The level of insulin released at terminal differentiation stage was tested by insulinrelease assay in vitro.RESULTS AND CONCLUSION: (1) There was no change in the imprinting state of the two kinds of mouse embryonicstem cells with normal and imprinted IGF-2 gene before and after differentiation. (2) In the induced cells, the expressionlevel of insulin in the SF1-1 group was higher than that in the SF1-G group, although there was no significant differencein the insulin release between the two kinds of cells. (3)The imprinting loss of IGF-2 gene may be related to theup-regulation of insulin mRNA expression in terminal cells during induced differentiation.
5.Surgical treatment of the cervical spine fractures combined with ankylosing spondylitis
Feng-Shan ZHANG ; Zhong-Jun LIU ; Zhong-Qiang CHEN ;
Chinese Journal of Orthopaedic Trauma 2004;0(12):-
Objective To study effects and other related problems of surgery for patients with cervical spine fracture and ankylosing spondylitis.Methods Twelve patients with cervical spine fracture and ankylosing spondylitis were treated with surgery from April 1986 to April 2004.All eases were studied retrospectively.They were followed up for an average of 67.5 months and their complete clinical data were kept.The neurological function was evaluated by ASIA (American Spinal Injury Association) impairment scale,and the image analyzing software (Image-Pro Plus 5.1) was used to measure the angles of superior and inferior vertebral bodies of the fusion segment at flexion and extension positions.The difference between flexion and extension angles,?,served as the parameter of interspinal movement.According to the definition of spinal fusion by FDA (Food and Drug Administration),the?≥4?was considered as nonfusion.Other related problems were discussed by descriptive study.Results The average improvement in the nine patients with neurological injury was 1.3 ASIA grades.The injured segments in 10 cases were treated with fusion,of whom nine were fused by internal fixation.The fusion rate was 100%.Three cases were scheduled to have laminoplasty,but two had to receive laminectomy instead because of intraoperative complete fracture at the hinged side.Ten patients were complicated by insufficient function of major organs preop- eratively,and endotracheal intubation was difficult to perform in eight cases.Postoperative complications occurred in three cases,but fortunately healed completely.There were no deaths or fatal complications.Conclusions The neurological function can be improved by surgery for patients with cervical spine fracture and ankylosing spondylitis. Although the cervical spine is instable for most of the patients,fusion with internal fixation is indicated and can be successful.Those who had preoperative systemic diseases are likely to suffer from postoperative complications.The difficult endotracheal intubation is a common intraoperative problem.
6.Data analysis of surveillance results of iodine deficiency disorders in Guangdong Province in 2011
Wen, ZHONG ; Li-ping, LIU ; Tong, YANG ; Li-feng, LIN ; Hai-shan, CHI ; Shan-hua, ZHANG
Chinese Journal of Endemiology 2013;32(6):677-681
Objective To assess the effectiveness of prevention program on iodine deficiency disorders and iodine nutritional status of residents in Guangdong Province.Methods Probability proportionate to size sampling(PPS) was employed in surveillance of iodine deficiency disorders.Thirty counties(cities,districts) were selected in Guangdong Province.In each county(city,district) one township(street) was selected; in each township (street) one primary school was selected and in each primary school 40 children aged 8-10 were chosen to examine their thyroid and to collect salt samples at their home for determination of salt iodine.Out of the 40 children,12 children were chosen to collect urine samples for determination of urinary iodine.From the primary schools chosen,40 grade 5 students were selected for intelligence quotient(IQ) test.In the nearby of the primary schools,3 townships(towns,street) were selected and in each township(town,street) 5 pregnant and 5 lactating women were selected to collect their urine samples for determination of urinary iodine.Type-B ultrasonic was used in measuring the thyroid volume.The iodine content of urine samples was measured by the method of arsenic and cerium catalysis spectrophotometry.The iodine content of salt was determined quantitatively with the titration method.IQ was tested by Chinese combined Raven's test.According to geographical location and the implementation of iodized salt,the effects of iodized salt on iodine deficiency disorders were analyzed in the plains and the Pearl River Delta Coastal region with mild iodine deficiency(iodized salt implementation region,referred to as the plains and the PRD),historical iodine deficiency areas (iodized implementation region) and the eastern and the western coastal areas of Guangdong(areas with non-iodized salt problem,referred to as the eastern and the western Guangdong).Results A total of 1200 children aged 8 to 10 were examined by type-B ultrasonic test,and goiter rate was 3.5% (42/1200).The differences of goiter rate between the plains and the PRD,the historical iodine deficiency areas and the eastern and the western Guangdong were statistically significant (x2 =6.6,P < 0.05).The goiter rate (6.1%) in the eastern and the western Guangdong was significantly higher than that of the plains and the PRD and the historical iodine deficiency areas (3.3%,2.0%,x2 =5.6,7.1,all P < 0.05).A total of 1200 salt samples were examined.The median and coefficient of variation of iodine in the salt were 31.0 mg/kg and 23.2%,respectively.Coverage of iodized salt was 97.5%(1170/1200) while 96.1%(1153/1200) of consumed iodized salt was qualified.The median urinary iodine of 1200 children aged 8-10 was 186.5 μg/L,and the differences of median urinary iodine between the plains and the PRD,the historical iodine deficiency areas and the eastern and the western Guangdong were statistically significant(x2 =5.9,P < 0.05).The median urinary iodine of the eastern and the western Guangdong(162.4 μg/L) was significantly lower than that of the plains and the PRD(207.5 μg/L,x2 =8.7,P < 0.01).The difference of median urinary iodine between the plains and the PRD,the historical iodine deficiency areas and the eastern and the western Guangdong was statistically significant(x2 =58.9,P< 0.01).The median urinary iodine of the eastern and the western Guangdong(109.6 μg/L) was significantly lower than that of the historical iodine deficiency areas and the plains and the PRD(152.9,155.2 μg/L,x2 =18.3,20.6,all P < 0.05).The mean IQ of the 1208 grade 5 students was 102.8 ± 14.3.The IQ of the plains and the PRD(104.3 ± 13.9) and the historical iodine deficiency areas(102.7 ± 14.3) was significantly higher than that of the eastern and the western Guangdong(100.3 ± 14.7,t =3.8,2.1,P< 0.01 orP< 0.05).Conclusions The goal of iodine deficiency disorders elimination is achieved as scheduled in Guangdong Province.The health level of general population has been improved significantly.Iodine nutrition is in the appropriate range (100-199 μg/L) in general population but low in pregnant women.The selling of non-iodized salt in the eastern and the western Guangdong Province should be followed closely.
7.Characterization of Aggregation-induced Toxicity by ?-synuclein in Yeast
Shan-Shan YU ; Yan LI ; Guan-Jun CHEN ; Wei-Feng LIU ;
Microbiology 1992;0(02):-
Parkinson's disease(PD) is the most common neurodegenerative movement disorder and ?-synuclein plays an important role in its development.Using the genetically tractable Saccharomyces cere-visiae as a model system,the phenotypic repercussions and potential mechanisms of ?-synuclein-induced cytotoxicity are characterized through modulating its expression level and other cellular factors.Aggrega-tion-induced toxicity is more dramatic upon elevated expression of ?-synuclein than that induced at moder-ate levels of expression.The induced toxicity is also enhanced by reagents such as dimethyl sulfoxide,which increase intracellular levels of phospholipid and membrane,as well as ferrous ions and hydrogen peroxide,both of which cause oxidative stresses in yeast cells.In contrast,over-expression of yeast homologue of hu-man chaperone DJ-1,YDR533C,markedly alleviate the inhibition of growth afflicted by exogenous expres-sion of ?-synuclein.Taken together,the data presented suggest a role for protein folding machinery together with quality control system in dealing with the aggregation of ?-synuclein.
8.Origin and trend of the upper eyelid artery in the eyelid region of a skull model after blood reperfusion: an anatomical observation
Xu-Feng LI ; Yuan-Yuan LIU ; Shan-Shan REN ; Ai WANG ; Xiao-Sheng LU
Chinese Journal of Tissue Engineering Research 2018;22(8):1241-1246
BACKGROUND: Upper eyelid flap grafting-related vessels such as superficial temporal artery, supratrochlear artery, supraorbital artery trunk are reported. Upper eyelid artery dissection is becoming more and more important for the surgery on the eyelid, but there is a lack of anatomical analysis of upper eyelid artery. OBJECTIVE: To measure the anatomical position of the upper eyelid artery in the eyelid region, and to provide anatomical basis for adjacent flap grafting. METHODS: Twenty adult skull specimens were dissected, and a reference coordinate system was made based on the inner canthus connection for the X axis, and the center line for the Y axis. The red lactoprene was injected into the skull model via common carotid artery.The locations A-E of the upper eyelid artery in the eyelid area were measured. RESULTS AND CONCLUSION: The upper eyelid artery in the eyelid area was mainly from the supratrochlear artery and the supraorbital artery, generally paralleling to the X axis. The upper eyelid branch originated from the supratrochlear artery was located at the projection of the inner canthus, with a total length of 24.50 mm, and a diameter of 0.51 mm, extended to the outer canthus and the diameter of the vessel gradually reduced. The upper eyelid branch originated from the supraorbital artery was located at pupil and inner canthus junction 1/2 projection. The total length of the blood vessels was about 23-24.6 mm, and the diameter of the blood vessels was (0.55±0.05) mm. In the current study, we obtained the surface projecting of upper eyelid artery in the eyelid area by establishing the skull model of blood perfusion, which provides an anatomic basis for upper eyelid flap grafting.
9.The effect of HIFU combined with nanoscale ultrasound molecular probes with simple virus thymidine kinase gene on angiogenesis in the nude mouse tumor
Hui XIE ; Shan LIU ; Hui ZENG ; Youwei LI ; Jin XU ; Xiong DING ; Changan LIU ; Feng LI
Chongqing Medicine 2017;46(6):725-728
Objective To observe the change of the protein and gene expression of hypoxia inducing factor-1α(HIF-1a) and vascular endothelial growth factor (VEGF) in the nude mouse tumor,which has been treated by HIFU combined with nanoscale ultrasound molecular probes with HSV1-TK gene microvascular density.Methods Sixty nude mice were implanted with HepG2 Cells to establish subcutaneous transplanted tumor.Divided this mice into six groups at random after treated by HIFU:MB+ HSV-TK+ GPC3 (group A),MB + HSV-TK (group B),HSV-TK +GPC3 (group C),HSV-TK (group D),MB + GPC3 (group E),PBS (group F).They were injected into the tail vein every after 3 days.Mice in group A,B,D and E were exposed to ultrasound by 2 W/cm2,1 MHz,5 mintues and 0.2 mL ganciclovi(GCV) was intraperitoneally injected at the first 48 hours after injection.After the treatment,immunohistoche were used to detect the microvascular density(MVD),Western blot and immunohistoche was employed to test the protein change of the VEGF and HIF-1α,Q-PCR was used to test the mRNA gene transcription of VEGF and HIF-1α in the tumor tissues.Results After 14 days,the protein expression of HIF-1α and VEGF in group A was significantly lower than that in group B,C,D,E and F (P<0.05),the MVD level in the tumor is also like this,and the difference is statistically significant.Conclusion Anoscale ultrasound molecular probes with HSV1-TK can reduce the the level of VEGF,MVD and HIF-1α in the tumor which has been treated by HIFU,so it can inhibit tumor growth and improve the therapeutic efficacy after HIFU treatment.
10.The study about the abilities of nursing master students in our country
Weiying SHAN ; Wenzhong AN ; Yali LIU ; Guiying FENG ; Zhong HE ; Huaping LIU
Chinese Journal of Practical Nursing 2006;0(12):-
Objective To explore the program of cultivating the nursing master students according to the present situation of China.Methods Delphi method was used in this study.Results In the type of scientific research,the order were the basic knowledge of nursing,clinical knowledge,research ability,ability of adapting society,teaching,management and the community nursing ability.In the type of clinic,the order were the basic knowledge of nursing,clinical knowledge,ability of adapting society,management,research ability,teaching and the community nursing ability.Conclusion According to the present situation of our country,the basic knowledge of nursing and the clinical skills should not be ignored.