1.RNAlater in parathyroid adenoma tissue storage
Jianguo ZHAO ; Zhao JIN ; Ya HU ; Quan LIAO ; Yupei ZHAO
Journal of Endocrine Surgery 2014;8(1):29-32
Objective To investigate different methods in parathyroid adenoma tissue storage and to provide theoretical support for standardized construction of parathyroid adenoma tissue data.Methods 22 samples of sporadic parathyroid adenoma in Beijing Union Hospital from Aug.2010 to Nov.2011 were collected.All cases obtained pathological diagnosis,among which 11 were kept in frozen nitrogen (group A)and the rest in RNAlater(group B).The ratio of OD 260/280 was measured by ultraviolet spectrophotometer.The integrity of RNA was examined by gel electrophoresis.The expression of β-actin was measured by Realtime-PCR technique.Results RNAlater group had no advantage than frozen nitrogen group in terms of the purity,concentration,and integrity of the extracted RNA(P >0.05),but had a higher expression level of the β-actin(P <0.05).Conclusion The method of RNAlater has no significant advantages compared with frozen nitrogen in terms of RNA prevention and RNA degradation,but is more effective in longer storage of samples.
2.A detailed criterion of BI-RADS assessment category based on US-BI-RADS: a preliminary study
Haina ZHAO ; Yulan PENG ; Honghao LUO ; Yushuang HE ; Ya JIN ; Pan YANG
Chinese Journal of Ultrasonography 2015;24(3):242-245
Objective To set up an objective criterion for BI-RADS assessment.Methods The breast sonograms of 1 938 female cases were retrospectively studied which had been confirmed by pathology from January to September 2011 and 2012 January.1 660 cases during 2011 were model cases,and the importance of each single feature in distinguishing between benign and malignant was obtained with Chi square and OR value.A detailed criterion of BI-RADS assessment category was set up based on US-BI-RADS.Results Eighteen of 24 lexicons were statistically significant distinguished between benign and malignant breast masses(P < 0.05).The lexicons were divided into major suspicious signs,middle ones and minor ones assigned 3 points,2 points and 1 point,and a scoring model was established as follows:Score =3 * (X1 + X2 + … + X6) + 2 * (X7 + X8 + … + X12) + (X9 + X10 + … + X18).Based on BI-RADS,the positive predictive value of model cases was 1.5%,6.9%,22.1%,62.5%,96.1% followed by category 3,4A,4B,4C and 5,and it was 1.4%,3.4%,21.1%,69.4%,92.7% in test cases.Conclusions The scoring model could be useful for BI-RADS final assessment more objectively,and could make it more convenient to predict the risk of breast cancer.
3.REPORT OF HLA DISTRIBUTION IN 2315 VOLUNTEER DONORS OF CHINESE BONE MARROW BANK FROM NORTHWEST CHINA
Tianbo JIN ; Ya GAO ; Hongbo ZHANG ; Junhai ZHAO ; Jianghua LAI ; Shuping LAI ; Shengbin LI
Journal of Pharmaceutical Analysis 2005;17(1):45-48
Objective To report the HLA data of volunteer donors of Chinese bank from Northwest China and characterize the distribution of HLA genes in Northwest China. Methods HLA-A, B antigens of 2315 volunteer donors were examined by the method of microlymphocytetoxicity (MLT) test .The antigen frequencies(AF) were assessed by directly counting; and based on that gene frequencies(GF) were calculated. HLA data from other population were collected and distribution characteristics were compared. With the raw data, Hardy-Weinberg equilibrium, statistical parameters of forensic medicine interest for HLA were computed. Results A total of 18 specific antigens were detected in HLA-A and the most frequent antigen was A2 . AF and GF were 0.5136 and 0.3026, respectively. A total of 42 specific antigens were detected in HLA-B and the most frequent antigen was A13. Its AF and GF were 0.1978 and 0.1044, respectively. The heterozygosity(H), polymorphism information content(PIC), discrimination power(DP) and probability of paternity exclusion (PPE) of HLA-A were 0.8215, 0.8212, 0.9356 and 0.7798 accordingly; while H,PIC, DP and PPE of HLA-B were 0.9322, 0.9322, 0.9878 and 0.9528. Conclusion The polymorphism of HLA-A,B genes is characteristic in Chinese. In this research, the genetic trait of HLA in 2315 volunteers is consistent with Northern Han population.
4.Study on excretion of pseudo-ginsenoside GQ.
Chun-Fang ZHAO ; Jin-Ping LIU ; Yan ZHAO ; Ping-Ya LI
China Journal of Chinese Materia Medica 2008;33(4):432-435
OBJECTIVETo determine the pseudo-ginsenoside GQ (PGQ) in rat bile, feces and urine, and to study on the excretion of pseudo-ginsenoside GQ.
METHODReverse phase high-performance liquid chromatography (RP-HPLC) method with an evaporative light-scattering detector (ELSD) was performed on Diamonsil C18 column (4.6 mm x 250 mm, 5 microm), and the mobile phase was consisted of methanol-water (24: 7) with flow rate of 1.0 mL x min(-1). ELSD parameters were set as follows: nitrogen gas pressure 3.0 bar, drift tube temperature 50 degrees C.
RESULTThe method fulfilled all the standard requirements of precision, accuracy and linearity. The main way of excretion of PGQ in rat administrated through sublingual vein was at the bile. The bile excretion ratio of PGQ was 41.60%, and feces excretion ratio was 9.97%. Only trace amount of PGQ was excreted in urine.
CONCLUSIONAlmost all unchanged PGQ was excreted in bile, feces and urine.
Animals ; Bile ; metabolism ; Chromatography, High Pressure Liquid ; Feces ; Female ; Ginsenosides ; administration & dosage ; metabolism ; pharmacokinetics ; urine ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction
5.Effects of exhaustive exercise-induced oxidative stress on red blood cell deformability.
Yan-Lian XIONG ; Yan-Lei XIONG ; Yao-Jin LI ; Fu-Zhou TANG ; Ruo-Feng WANG ; Ya-Jin ZHAO ; Xiang WANG
Chinese Journal of Applied Physiology 2014;30(4):289-293
OBJECTIVEThe aim of the present study is to explore the effects of exhaustive exercise-induced oxidative stress on the antioxidant capacity and diformability of rat red blood cells.
METHODSRats were divided into three group (n = 10): sedentary control (C), exhaustive running exercise (ERE) and moderate running exercise (MRE) groups. Animals in the ERE group started treadmill running at a speed of 20 m/min speed with a 5% gradient, and reached a speed of 25 m/min with gradient 15% in 20 min. Running was continued until exhaustion. MRE group rats running at a speed of 20 m/min with a 5% gradient for 40 min. The levels of free thiol in erythrocyte membrane protein, lipidperoxidation levels and membrane protein components were analyzed. The red blood cell deformability of different groups was also observed.
RESULTSThe results showed that red blood cells were damaged by severe oxidative stress and the anti-oxidative capacity decreased significantly under exhaustive exercise conditions. Besides, lipid peroxidation and protein sulfhydryl cross-link based clustering of membrane were found after exhaustive exercise, and polymers high molecular weight (HMW) was formed. The elongation index (EI) was found to decline significantly in the ERE group compared with the C and MRE groups under shear stress (control group, 0.41 +/- 0.01 at 3 Pa and 0.571 +/- 0.008 at 30 Pa; ERE group, 0.314 +/- 0.013 at 3 Pa and 0.534 +/- 0.009 at 30 Pa; P < 0.05 and P < 0.01, respectively).
CONCLUSIONThese exercise-induced oxidative injure result in a significant decrease in deformability of rat erythrocytes, which in turn leads to dysfunction in the microcirculatory.
Animals ; Disease Models, Animal ; Erythrocyte Deformability ; Fatigue ; metabolism ; physiopathology ; Male ; Oxidative Stress ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley
6.Effect of Xuebijing injection on TLR4--NF-kappaB--IL-1beta pathway of myocardial hypoxia/reoxygenation in rats.
Ya-Kun LIU ; Lin-Jing HUANG ; Shan ZHAO ; Wei LIN ; Jin-Bo HE ; Lei YING ; Xin YOU ; Wan-Tie WANG
Chinese Journal of Applied Physiology 2014;30(1):55-59
OBJECTIVETo investigate the role of Xuebijing injection(XBJI, traditional Chinese medicine), in inhibiting TLR4--NF-kappaB--IL-1beta pathway of myocardial hypoxia/reoxygenation in rats.
METHODSThirty six male SD rats (280 +/- 30) g were randomly divided into six groups (n = 6): normal group (N group), balanced perfusion group (BP group), model group (M group), low dose XBJI group (XBJI(L) group), middle dose XBJI group (XBJI(M) group), high dose XBJI group (XBJI(H) group). By Langendorff isolated heart perfusion device to establish the model of myocardial hypoxia/reoxygenation in rats. ELISA was used to detect the concentration of interleukin-1beta (IL-1beta); Western blot was used to detect the expression of nuclear factor kappa B p65 (NF-kappaB p65) protein and toll like receptor 4 (TLR4) protein; and RT-PCR to determine the expression of NF-kappaB p65 mRNA and TLR4 mRNA;To observe microstructure changes of hypoxia/reoxygenation myocardial by light microscopy.
RESULTSCompared with M group, the IL-1beta concentration, NF-kappaB p65 and TLR4 protein,NF-kappaB p65 and TLR4 mRNA of XBJIL group, XBJI(M) group, XBJI(H) group expression decreased in varying degrees,and decreased most obviously all in XBJI(M) group (P < 0.05, P < 0.01); Myocardical structural damage was serious in M group, and improved after treatment XBJI, the most obvious was the XBJI(M).
CONCLUSIONDifferent dose of XBJI can inhibit TLR4--NF-kappaB--IL-1beta signal transduction pathway and reduce several inflammatory reaction after myocardial hypoxia/reoxygenation injury, the 4 ml/100 ml of XBJI is the best.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Heart ; drug effects ; Inflammation ; Interleukin-1beta ; metabolism ; Male ; Myocardium ; pathology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism
7.Effect of bear bile powder on STAT3 pathway in hepatocellular carcinoma xenograft.
Jin-Yan ZHAO ; Li-Ya LIU ; A-Ling SHEN ; Wei LIN ; Zhi-Yun CAO ; Qun-Chuan ZHUANG ; Zhen-Feng HONG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(8):976-981
OBJECTIVETo observe the effect of bear bile powder (BBP) on the STAT3 pathway and its downstream target genes of nude mice hepatocellular carcinoma (HCC) xenograft, and to explore its mechanism for treating HCC.
METHODSThe subcutaneous xenograft model was established using HepG2 cells. When the subcutaneous transplanted tumor was formed, naked mice were randomly divided into two groups, the BBP group and the control group. Mice in the BBP group were administered with BBP by gastrogavage, once daily for 3 consecutive weeks, while mice in the control group were administered with normal saline by gastrogavage, once daily for 3 consecutive weeks. The body weight and the tumor volume were measured once per week. By the end of medication, the tumor weight was weighed and the tumor inhibition ratio calculated. The apoptosis of the tumor tissue was detected by TdT-mediated dUTP nick end labeling (TUNEL). The expression of Bcl2-associated X protein (Bax), B cell lymphoma/eukemina-2 (Bcl-2), cyclin-dependent protein kinase (CDK4), cyclinD1 were detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of signal transducers and transcription activators 3 (p-STAT3), proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, CDK4, and cyclinD1 were determined by immunohistochemistry.
RESULTSBBP could inhibit the tumor volume and tumor weight, showing statistical difference when compared with the control group (P < 0.01). Results of TUNEL showed that BBP could significantly induce the apoptosis of hepatoma carcinoma cells. Results of RT-PCR showed that BBP could up-regulate the expression of Bax and down-regulate mRNA expression of Bcl-2, CDK4, and cyclinD1. Immunohistochemical results showed that BBP could up-regulate the expression of Bax and inhibit the protein expression of p-STAT3, PCNA, Bcl-2, CDK4, and cyclinD1.
CONCLUSIONBBP could induce the apoptosis of hepatoma carcinoma cells and inhibit their proliferation by regulating STAT3 pathway.
Animals ; Bile ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Ursidae ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism
8.Expression of STEAP4 Gene during the Period of Human Preadipocyte Differentiation
xiao-hui, CHEN ; ya-ping, ZHAO ; chun-lin, GAO ; chun-mei, ZHANG ; chun, ZHU ; jin-gai, ZHU ; xi-rong, GUO
Journal of Applied Clinical Pediatrics 2004;0(07):-
Objective To observe the expression of STEAP4 gene(a novel obesity-related gene) during the period of human preadipocyte differentiation and to explore the relationship between the STEAP4 gene expression and adipocytes differentiation,adipogenesis.Methods Human preadipocytes were cultured and differentiated into the matured adipocytes in vitro.Adipocytes morphology and lipid accumulation were observed during this process.Total RNA was extracted from adipocytes at various time points (preadipocyte,Day 0,Day 4,Day 6,Day 8,Day 11,Day 14,and Day 17) and the level of STEAP4 mRNA expression was measured by fluorescent real-time quantitative reverse transcriptase-polyme-rase chain reaction(RT-PCR).Results The level of STEAP4 mRNA expression remained high in preadipocytes.In the presence of differentiation medium (Day 4),there was a transient upregulation in the expression of STEAP4 gene.After that,with the human preadipocytes being differentiated into matured adipocytes,the expression of STEAP4 mRNA was downregulated and reached the lowest level in fully differentiated adipocytes.There was a significant difference between any 2 detected phases in the level of STEAP4 mRNA expression (Pa
9.The Research on Stability of an Isolate of Riemerrella anatipestifer
Ya-Ni SUN ; Qin ZHAO ; Shi-Jin JIANG ; Xing-Xiao ZHANG ; Yi-Bo KONG ; Xiu-Li WEI ;
Microbiology 2008;0(07):-
The isolate GN52 of Riemerrella anatipestifer was passaged on the Martin Medium successively according to the optimum condition. The experiments included Gram staining, biochemical test, drug sensitivity test and animal experiments were carried out on the bacteria of 3rd, 11th, 21st, 31st, 41st, 51st and 61st generations. It indicated that the bacterial morphs, biochemical character, drug resistance of the strain had no obvious change, but the virulence showed a trend of reduction.
10.Mechanism of BCL2L2-PABPN1 expression induced by sodium arsenite and its metabolites in 16HBE cells
SHI Ya YIN Jin yao WU Jiang JIANG Cheng lan ZHAO Rui huan ZHOU Qian HE Yue feng
China Occupational Medicine 2022;49(05):522-
Objective - - (BCL2L2)- ( )
To investigate the differential expression of the fusion gene BCL 2 like protein 2 poly A
(PABPN1) ( )
binding protein nuclear 1 induced by sodium arsenite SA and its methylated metabolites in 16HBE cells and the
Methods ) ,
related mechanism. i The 16HBE cells exposed to SA at concentrations of 1.5 3.0 and 4.5 µmol/L were set as
-, - -
low medium and high dose arsenic exposure groups. The 16HBE cells exposed to 4.5 µmol/L monomethylarsonic acid
( ), ( ) ,
MMA dimethylarsonic acid DMA and SA were set as MMA group DMA group and SA group. The 16HBE cells without
, BCL2L2-PABPN1
toxic stimulation were set as control group. After the cells were cultured for 48 hours the expression of was
- ( - ) ) ( )
detected by quantitative real time polymerase chain reaction qRT PCR . ii Two small interfering RNA siRNA silencing
基金项目:国家自然科学基金( ); 年云南省科技厅昆明医科大学应用基础研究联合专项面上项目
82160607 2021
( )
202101AY070001-054
作者简介:施雅( —),女,在读大学本科生,主要从事劳动卫生与环境卫生学研究;尹锦瑶( —),女,在读劳动卫生与环境卫
2001 1995
生学硕士研究生,主要从事劳动卫生与环境卫生学研究;施雅和尹锦瑶为共同第一作者
通讯作者:何越峰教授,博士研究生导师,- :
E mail heyuefeng@kmmu.edu.cn中国职业医学 年 月第 卷第 期 , , , · ·
2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 523
BCL2L2-PABPN1, -
fragments were designed and transfected into 16HBE cells to knockdown which were set as siRNA 1 group
- - BCL2L2-PABPN1
and siRNA 2 group. Non transfected control group without knockdown of transfection was set up. After
, BCL2L2-PABPN1 -
culturing for 48 hours the expression level of in the three groups of cells was detected by qRT PCR. The cell
-
survival rate and early apoptosis rate were detected by MTS method and JC 1 mitochondrial membrane potential detection
, ( ) ,
method respectively. The apoptosis was detected by Hoechest33342/propidium iodide PI double staining and the expression
- Results )
level of P53 signaling pathway related proteins was detected by Western blotting. i The relative expression of
BCL2L2-PABPN1 (P ) BCL2L2-
in 16HBE cells increased with the increasing SA doses <0.01 . The relative expression of
PABPN1 - , - -
in high dose arsenic exposure was higher than that in control group low dose and medium dose arsenic exposure
( P ) BCL2L2-PABPN1 ,
groups all <0.05 . The relative expression of in SA group was higher than those in control group MMA
( P ) BCL2L2-PABPN1
group and DMA group all <0.05 . The relative expression of showed no significant difference between
, ( P ) ) BCL2L2-PABPN1
control group MMA group and DMA group all >0.05 . ii The relative expression levels of and cell
- - - ( P )
survival rate in siRNA 1 group and siRNA 2 group were lower than those in non transfected control group all <0.05 .
, (P )
However there was no significant difference in the early apoptosis rate among the three groups >0.05 . The results of
-
Hoechest33342/PI double staining showed that the number of nuclear shrinkage and early apoptotic cells in siRNA 1 group and
- - , -
siRNA 2 group was higher than that in non transfected control group. The relative protein expression levels of P53 phospho
, - - , - - ( P )
p53 BCL 2 associated death promoter P21 and cytochrome C in siRNA 1 group and siRNA 2 group were higher all <0.05 ,
- - P
and the relative protein expression levels of P53 up regulated modulator of apoptosis were lower (all <0.05), when compared
- Conclusion
with the non transfected control group. SA may block the apoptosis of 16HBE cells by inducing the expression of
BCL2L2-PABPN1
fusion gene . The mechanism may be related to the activation of P53 signaling pathway. The SA methylated
BCL2L2-PABPN1 BCL2L2-PABPN1 -
metabolites MMD and DMA had no effect on the expression of . may affect anti apoptosis
BCL2L2 PABPN1
through affecting the synergistic effect of and genes.