1.Fatty acid participates in up-regulation of diabetes on function and expression of CYP1A2
Nan HU ; Yan JIANG ; Rui HAN ; Qing QIAN ; Sulan ZOU
Chinese Pharmacological Bulletin 2017;33(2):249-254
Aim To investigate the mechanism of dia-betes changing the hepatic CYP1 A2 through in vitro cell culture study.Methods The function of CYP1 A2 in HepG2 and Fa2N-4 cells were evaluated by determi-ning the level of phenacetin metabolism,and the mR-NA expression of CYP1 A2 in cells was detected by real time PCR.HepG2 cells were co-cultured with serum of diabetic rats(type 1 and type 2)and normal rats,then the CYP1 A2 function in cells were evaluated.Then, the HepG2 and Fa2N-4 cells were co-cultured with a series of concentrations of saturated (including palmitic acid and stearic acid)and unsaturated fatty acids(in-cluding oleic acid and linoleic acid)for 48 h,and the function and expression of CYP1 A2 in the cells were compared.Results It was found that the activities of CYP1 A2 were higher in cells incubated with diabetic serum of both type.All high concentration of fatty acids could increase the function and expression of CYP1 A2 in both HepG2 and Fa2N-4 cells.Conclusion It is speculated that the abnormal level of fatty acids under diabetic state might be part of the reasons why diabetes change the hepatic CYP1 A2,which provides the basis for future study.
2.DETERMINATION OF TOTAL BODY FAT BY WATER DISPLACEMENT METHOD
Qing-Hui YANG ; Bing-Zhang DUAN ; Ya-Nan JIANG ;
Acta Nutrimenta Sinica 1956;0(02):-
Body fat of 12 male adults were measured by water displacement me-thod(density method) at every morning for 5 successive days. The standard deviation of single observation was 0.29kg calculated by mean residual lung volume method. It was significantly lower than the value (0.5kg) calculated by the ordinary method (p
3.Application of Ion Torrent PGM™ System in Detection of Fetal DNA in Maternal Plasma.
Ya-nan LIU ; Xue-ying ZHAO ; Yuan PING ; Qing-wen XU ; Jiang-ping HUANG ; Kai-nan ZOU ; Huai-gu ZHOU
Journal of Forensic Medicine 2015;31(6):432-435
OBJECTIVE:
To explore the feasibility of detecting of Y-STR of fetal DNA in maternal plasma using Ion Torrent PGM™ System.
METHODS:
A total of 16 fetal DNA samples from maternal plasmas (8 cases from 38 weeks gestational age and 8 ones from 12 weeks) were prepared and a multiplex assay with 7 STR loci (DYS390, DYS391, DYS393, DYS438, DYS437, DYS456, DYS635) was designed for multiplex-PCR amplification. Using Ion Torrent PGM™ System, the results of Y-STR sequences and capillary electrophoresis were obtained and compared.
RESULTS:
Y-STR specific alleles were detected in the maternal plasma of all the pregnant women having male babies of second and third trimester, which were higher than that detected by capillary electrophoresis. Consistent Y-STR genotypes were observed between fetal DNA from maternal plasma and genomic DNA from the newborn babies.
CONCLUSION
Based on Ion Torrent PGM™ System, the prenatal Y-STR detection method may provide a high-sensitive and high-throughput choice for prenatal STR detection in forensic testing.
Alleles
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Chromosomes, Human, Y/genetics*
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DNA/blood*
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Family
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Female
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Fetal Blood/chemistry*
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Genotype
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Haplotypes
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Humans
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Male
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Polymerase Chain Reaction
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Polymorphism, Genetic
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Pregnancy
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Sensitivity and Specificity
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Sex Determination Analysis
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Tandem Repeat Sequences/genetics*
4.Clinical features and correlation between radiographic parameters and incidence of calcaneal spur
Qing ZHANG ; Nan JIANG ; Weiran HU ; Zewei YU ; Xiang ZHANG ; Bin YU
Chinese Journal of Orthopaedic Trauma 2016;18(6):487-492
Objective To analyze the clinical features and the correlation between radiographic parameters and incidence of calcaneal spur in the patients from Nanfang Hospital,Southern Medical University,China.Methods Three experienced observers independently used the image acquisition and transmission system (PACS) to collect the data of lateral and axial X-ray images of calcaneus or ankle joint in neutral position from the patients with calcaneal spur and normal controls who had undergone radiological examination in Nanfang Hospital,Southern Medical University from July 2014 through December 2015.Ten radiological parameters of the foot (B(o)hler angle,Gissane angle,calcaneal inclination angle,talocalcaneal angle,talus horizontal angle,posterior facet inclination angle,calcaneal length,height of the posterior facet,absolute foot height,and calcaneal width) were measured in both the patients and normal controls.The location,morphology and length of calcaneal spurs were compared between genders,sides and age groups.Results A total of 216 parpatients were included in the study.Female patients were more than male ones,simple plantar spurs more than simple achilles tendon ones,type B spurs more than type A ones,the length of achilles tendon spurs larger than that of plantar ones,female plantar spurs more than males ones,and the length of right foot plantar spurs larger than that of left foot ones.All the differences above were statistically significant (P < 0.05).There were no significant differences in the location,morphology or length of calcaneal spurs between the age group of ≤ 60 years old and the age group of > 60 years old (P > 0.05).The incidence of calcaneal spur were significantly correlated to Gissane angle (P =0.000,OR =0.944,95% CI 0.917-0.973),posterior facet inclination angle (P=0.017,OR=0.957,95% CI 0.924-0.992) and height of the posterior facet (P =0.007,OR =0.933,95% CI O.886-0.981).Conclusions Calcaneal spur favored more females than males.Plantar spurs were more common than Achilles ones.Plantar spurs of Type B were more common than those of Type A.Achilles spurs were longer than plantar ones.More females suffered plantar spur than males.Right foot spurs were longer than left foot ones.Age had no significant influence on the spur characteristics.The incidence of spur might have been related to the Gissane angle,posterior facet inclination angle and height of the posterior facet of the foot.
5.Effect evaluation of bedside ultrasound monitoring of left ventricular functional parameters combined with clinical indicators on veno-arterial extracorporeal membrane oxygenation
Renfeng YI ; Juan GUO ; Qing ZHOU ; Hongning SONG ; Yanxiang ZHOU ; Nan JIANG ; Xue YAO ; Ruiqiang GUO
Chinese Critical Care Medicine 2021;33(3):329-333
Objective:To explore the monitoring value of left ventricular functional parameters obtained by bedside ultrasound combined with clinically relevant indicators in patients with veno-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:A retrospective study was conducted. A total of 24 patients receiving VA-ECMO adjuvant support in Renmin Hospital of Wuhan University from June 2018 to January 2020 were selected. The bedside ultrasound was performed on the first day of ECMO support, the day before weaning, the clinical indicators before weaning were obtained. The differences in clinical indicators and the left ventricular functional parameters between the two groups of whether weaning successfully were compared; univariate Logistic regression analysis was used to screen out the related factors affecting weaning.Results:Sixteen patients were successful weaned and 8 patients failed. Compared with the weaning failure group, patients in the weaning success group required less continuous renal replacement therapy (CRRT, cases: 4 vs. 6, P < 0.05), mean arterial pressure (MAP) before weaning was higher [mmHg (1 mmHg = 0.133 kPa): 84.64±9.55 vs. 62.30±8.79, P < 0.05], and the pulse oxygen saturation (SpO 2) was also higher (0.966±0.670 vs. 0.866±0.061, P < 0.05), while vasoactive-inotropic score (VIS), serum creatinine (SCr) and serum lactic acid (Lac) were lower [VIS score: 7.27±1.42 vs. 16.93±8.52, SCr (μmol/L): 123.60±83.64 vs. 213.10±117.39, Lac (mmol/L): 1.94±0.91 vs. 5.62±5.48, all P < 0.05]. Univariate Logistic regression analysis showed that the MAP, VIS, SCr, Lac, SpO 2 before weaning were the related factors affecting weaning [odds ratio ( OR) were 0.306, -0.740, -0.011, -0.632, -4.069; 95% confidence interval (95% CI) were 1.065-1.732, 0.235-0.899, 0.979-0.999, 0.285-0.992 and 0.001-0.208; P values were 0.014, 0.022, 0.038, 0.047, 0.002]. In the weaning success group, left ventricular ejection fraction (LVEF), velocity of mitralannulus in systolic (LatSa), maximum flow velocity of aortic valve (AV-Vmax), velocity-time integral (VTI), left ventricular global longitudinal strain (LVGLS), left ventricular global longitudinal strain rate (LVGLSr) were all increased on the day before ECMO weaning compared with the first day of ECMO support [LVEF: 0.40±0.05 vs. 0.28±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.62±1.02, AV-Vmax (cm/s): 104.81±33.98 vs. 64.44±16.85, VTI (cm): 14.56±3.11 vs. 7.96±1.98, LVGLS: (-8.95±2.59)% vs. (-5.26±1.28)%, LVGLSr (1/s): -0.48±0.11 vs. -0.29±0.09], whereas the ECMO flow was significantly reduced (L/min: 1.46±0.47 vs. 2.64±0.31), the differences were statistically significant (all P < 0.05). There was no significant difference in left ventricular functional parameters between the first day of ECMO support and the day before ECMO weaning in the weaning failure group. Compared with the weaning failure group, the weaning success group had higher LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr on the day before ECMO weaning [LVEF: 0.40±0.05 vs. 0.26±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.31±1.03, AV-Vmax (cm/s): 104.81±33.98 vs. 67.67±18.46, VTI (cm): 14.56±3.11 vs. 7.75±2.77, LVGLS: (-8.95±2.59)% vs. (-4.81±1.81)%, LVGLSr (1/s): -0.48±0.11 vs. -0.30±0.10, all P < 0.05] and lower ECMO flow (L/min: 1.46±0.47 vs. 2.20±0.62, P < 0.05). Conclusion:Bedside echocardiographic left ventricular function parameters (LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr) combined with clinical indicators (MAP, VIS, SCr, Lac, SpO 2) were helpful to evaluate the therapeutic effect of patients receiving VA-ECMO support and can provide important guiding value in the selection of VA-ECMO weaning timing and the judgment of prognosis.
6.Experiment study on the transfection of exogenous genes promoted by ultrasound-targeted microbubbles combined with a peptide nucleic acid binding nuclear localization signal
Nan JIANG ; Qian CHEN ; Bo HU ; Qing ZHOU ; Sheng CAO ; Chuangli FENG ; Ruiqiang GUO
Chinese Journal of Ultrasonography 2017;26(5):442-447
Objective To increase the transfection of EGFP-N3 plasmids into 293T cells using ultrasound-targeted microbubbles delivery(UTMD) mediated a peptide nucleic acid (PNA) binding nuclear localization signal (NLS).Methods Antibody-targeted microbubbles were used in the experiments which can specifically recognize the SV40T antigen receptor.The SV40T antigen receptors were expressed on the membranes of 293T cells.The PNA containing the NLS were inserted in the EGFP-N3 plasmid DNA,which increased nuclear localization.Ultrasound-targeted microbubble delivery (UTMD) and the PNA binding NLS were utilized to improve the cytoplasmic import of plasmids and the nuclear intake of the plasmid from the cytoplasm,respectively.The study was divided into five groups:Contrast (group A),Common microbubble + DNA (group B),Antibody-targeted microbubbles + DNA (group C),Common microbubbles + NLS-PNA-DNA (group D),Antibody-targeted microbubbles + NLS-PNA-DNA (group E).Fluorescence microscope was used to observe the fluorescent light in each group;flow cytometry to test the transfection;RT-PCR and Western blot to detect genes' mRNA and protein expression level.Results Ultrasound and antibody-targeted microbubble delivery (UTMD) significantly enhanced the cytoplasmic intake of exogenous genes and maintained high cell viability(>80%).Fluorescent microscope showed that the quantities of green fluorescence in cells were increased successfully.The transfection results of flow cytometry were 0,(9.30 ± 0.46)%,(26.46 ± 2.01)%,(29.54 ± 0.62)%,(45.72 ± 1.86)%,respectively,and the differences were statistically significant(P <0.05).The relative mRNA and protein expression in group E were greater than those in group C and D respectively (P <0.05).Conclusions UTMD combined with antibody-targeted microbubbles and a PNA binding NLS plasmid can significantly improve transfection efficiency of exogenous genes by enhancing both cytoplasmic and nuclear DNA import.
7.Experimental study of amniotic lacrimal duct stent used to prevent dry eye of castrated rabbits
Mingyang MA ; Qing YUAN ; Qi LIU ; Kangcheng LIU ; Peiwen ZHU ; Honghua KANG ; Nan JIANG ; Lei YE ; Chonggang PEI ; Yi SHAO
Recent Advances in Ophthalmology 2017;37(8):709-713
Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.
8.Protective effect of water soluble CoQ10 on rotenone-induced apoptosis in PC12 cells
Yu-Min JIANG ; Hai-Ning LI ; Shao-Qing LIN ; Yan-Yan CHEN ; Jing AN ; Chun-Huan MA ; Nan-Nan HUAN ; Jiang CHENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2018;39(4):514-518
Objective To investigate the protective effect and the underlying mechanism of water soluble coenzyme Q10 (CoQ10)against rotenone induced injury on PC12 cells model.Methods PC12 cells were cultured with rotenone,water-soluble CoQ1 0 was added to the culture media 3 hours prior to the rotenone incubation.We determined cell viability by CCK8;reactive oxygen species (ROS)was detected by spectrophotometer;and Bcl-2, Bax,active Caspase-3,Caspase-9 and apoptosis-inducing factor (AIF)were measured by Western blotting after 24-hour rotenone incubation.Results After the treatment by rotenone,cell viability decreased significantly (P<0.01)and ROS level increased (P<0.01).CoQ10 could improve PC12 cell viability (P<0.01)and reduce the level of ROS (P<0.01).Western blotting experiments showed that CoQ10 could reduce rotenone-induced Caspase-9 (P<0.05),active Caspase-3 (P<0.05)and Bax (P<0.01)expressions,increase the expression of Bcl-2 (P<0.01),and prevent nuclear translocation of AIF (P<0.05).Conclusion CoQ10 has a protective effect on rotenone-induced apoptosis in PC12 cells,the mechanism of which may be through scavenging ROS in cells;decreasing caspase-9 ,active caspase-3 and Bax expressions;and increasing the expression of Bcl-2 ;and preventing AIF nuclear translocation.
9.Sequence analysis for genes encoding nucleoprotein and envelope protein of a new human coronavirus NL63 identified from a pediatric patient in Beijing by bioinformatics.
Jiang-feng XING ; Ru-nan ZHU ; Yuan QIAN ; Lin-qing ZHAO ; Jie DENG ; Fang WANG ; Yu SUN
Chinese Journal of Virology 2007;23(4):245-251
The aim of this study was to characterize the N and E protein encoding genes of a new human coronavirus (HCoV-NL63) which was identified from one of the clinical specimens (BJ8081) collected from a 12 years-old patient with acute respiratory infection in Beijing. The complete N and E gene sequences of HCoV-NL63 were amplified from clinical sample by RT-PCR, then were cloned into the pCF-T and pUCm-T vectors respectively and sequenced. The complete sequences of N and E genes were submitted to GenBank by Sequin and compared with N and E genes of prototype HCoV-NL63 and the other coronaviruses published in GenBank. The secondary structure and the characteristics of sample BJ8081 N and E proteins were predicted by bioinformatics. It was indicated that the N and E genes amplified from sample BJ8081 were 1134 bp and 234 bp in length and the predicted proteins including 377 amino acids and 77 amino acids, respectively. The data suggested that the region of amino acids 78-85 within N protein probably was the conserved region for all coronaviruses identified so far including HCoV-NL63. The region of amino acids 15-37 for E protein was probably the transmembrane domain. In conclusion, the recombinant plasmids pCF-T-8081 N and pUCm-T-8081 E were successfully constructed and sequenced, and the data predicted by bioinformatics are helpful for the further analysis of HCoV-NL63.
Amino Acid Sequence
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Child
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China
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Computational Biology
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methods
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Coronavirus
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classification
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genetics
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metabolism
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Coronavirus Infections
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virology
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Humans
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Molecular Sequence Data
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Nucleocapsid Proteins
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chemistry
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genetics
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metabolism
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Phylogeny
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Protein Structure, Secondary
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Viral Envelope Proteins
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chemistry
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genetics
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metabolism
10.Effect of small interfering RNA targeting Rac1 gene on colony formation of SW480 cells in vitro.
Qing-zhen NAN ; Lei GAO ; Bing XIAO ; Zhen-shu ZHANG ; Bo JIANG
Journal of Southern Medical University 2010;30(6):1339-1342
OBJECTIVETo construct a vector expressing small interfering RNA (siRNA) against Rac1 gene and observe its effect on soft agar colony formation of SW480 cells in vitro.
METHODSOligos of 64 base pairs for hairpin RNA targeting Rac1 were chemically synthesized and annealed. The siRNA constructs for Rac1, produced by inserting the annealed oligos into the downstream of H1 promoter of linearized pSUPER, were confirmed by restriction digestion and DNA sequencing. The constructed Rac1-siRNA was transfected into SW480 cells and Western blotting was performed to assess the expression and interference efficiency of siRNAs against Rac1.The soft agar colony formation assay was used to study the effect of Rac1 gene silencing on SW480 cells.
RESULTSRestriction digestion and DNA sequencing showed that the siRNA targeting Rac1 gene was successfully constructed. The siRNA could effectively down-regulate the expression of Rac1 in SW480 cells. Soft agar colony formation assay showed that the colony number and diameter of SW480 cells was reduced after siRNA transfection.
CONCLUSIONA vector expressing hairpin RNA against Rac1 gene are successfully produced, which significantly reduces the colony numbers and size of SW480 cells in vitro, suggesting that Rac1 plays an important role in the growth of colorectal cancer in vitro.
Base Sequence ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Down-Regulation ; Humans ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; rac1 GTP-Binding Protein ; genetics