1.Effects of lncRNA MIAT on cell proliferation of human non-small-cell lung carcinoma
Zhen PEI ; Xiao-Lei HUO ; Xiang-Yang TIAN ; Yi-Qiang ZHANG ; Jian-Tao JIA ; Ling-Na HAN
Chinese Journal of Pathophysiology 2018;34(4):592-598
AIM:To explore the expression level of long non-coding RNA(lncRNA)myocardial infarction-as-sociated transcript(MIAT)in the tissues and cells of non-small-cell lung carcinoma(NSCLC), and to investigate the effect of MIAT on the function of NSCLC cell line.METHODS:Bioinformatic data in microarray dataset GSE19804 from Gene Expression Omnibus(GEO)were collected for analyzing the difference expression of MIAT between NSCLC tissues and normal lung tissues.Clinical and prognostic data in microarray dataset GSE 30219 from GEO were also collected for an-alyzing the correlation between the expression level of MIAT and the survival time of NSCLC patients.qPCR was applied to detect the expression of MIAT in 25 paired tumor tissues and corresponding adjacent normal tissues,normal lung epithelial HBE cell line and NSCLC A549,NCI-H266 and NCI-H1299 cell lines.The specific small interfering RNA for MIAT(si-MIAT group)or negative control sequence(si-NC group)was transfected into A549 cells,and flow cytometry,colony for-mation experiment and CCK-8 assay were employed to detect the proliferation of the cells in the 2 groups.The expression levels of cyclin D1 and cyclin-dependent kinase inhibitor 1A(CDKN1A)in the 2 groups were determined by qPCR and Western blot.RESULTS:In the GEO dataset GSE19804,the expression of MIAT in NSCLC tissues was significantly ele-vated compared with normal lung tissues(P<0.05).In the GEO dataset GSE30219,the overall survival time was signifi-cantly shorter in the patients with high expression of MIAT than the patients with low expression of MIAT(P<0.05).Fur-thermore,the levels of MIAT in both NSCLC tissues and cells were higher than those in adjacent normal tissues and normal cells(P<0.05).Compared with si-NC group,lower MIAT level,cell viability and cell colony number in si-MIAT group with statistical significance were observed(P<0.05).Meanwhile, compared with si-NC group, the expression of cyclin D1 in si-MIAT group was significantly decreased(P<0.05),and inversely,the expression of CDKN1A in si-MIAT group was significantly increased(P<0.05).CONCLUSION:There is high expression of MIAT in NSCLC tissues and NSCLC cells,and knockdown of MIAT expression inhibits NSCLC cell proliferation, which provides a potential target of targeted therapy for NSCLC.
2.T Helper 1 and T Helper 2 Cytokines Differentially Modulate Expression of Filaggrin and its Processing Proteases in Human Keratinocytes.
Zheng-Hong DI ; Lei MA ; Rui-Qun QI ; Xiao-Dong SUN ; Wei HUO ; Li ZHANG ; Ya-Ni LYU ; Yu-Xiao HONG ; Hong-Duo CHEN ; Xing-Hua GAO
Chinese Medical Journal 2016;129(3):295-303
BACKGROUNDAtopic dermatitis (AD) is characterized by defective skin barrier and imbalance in T helper 1/T helper 2 (Th1/Th2) cytokine expression. Filaggrin (FLG) is the key protein to maintaining skin barrier function. Recent studies indicated that Th1/Th2 cytokines influence FLG expression in keratinocytes. However, the role of Th1/Th2 cytokines on FLG processing is not substantially documented. Our aim was to investigate the impact of Th1/Th2 cytokines on FLG processing.
METHODSHaCaT cells and normal human keratinocytes were cultured in low and high calcium media and stimulated by either interleukin (IL)-4, 13 or interferon-γ (IFN-γ). FLG, its major processing proteases and key protease inhibitor lymphoepithelial Kazal-type-related inhibitor (LEKTI) were measured by both real-time quantitative polymerase chain reaction and Western blotting. Their expression was also evaluated in acute and chronic AD lesions by immunohistochemistry.
RESULTSIL-4/13 significantly reduced, while IFN-γ significantly up-regulated FLG expression. IL-4/13 significantly increased, whereas IFN-γ significantly decreased the expression of kallikreins 5 and 7, matriptase and channel-activating serine protease 1. On the contrary, IL-4/13 significantly decreased, while IFN-γ increased the expression of LEKTI and caspase-14. Similar trends were observed in AD lesions.
CONCLUSIONSOur results suggested that Th1/Th2 cytokines differentially regulated the expression of major FLG processing enzymes. The imbalance between Th1 and Th2 polarized immune response seems to extend to FLG homeostasis, through the network of FLG processing enzymes.
Caspase 14 ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Dermatitis, Atopic ; metabolism ; Humans ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Intermediate Filament Proteins ; metabolism ; Keratinocytes ; enzymology ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; metabolism ; Serine Peptidase Inhibitor Kazal-Type 5 ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism
3.Expression of p53 isoforms in renal cell carcinoma.
Wei SONG ; Shi-wei HUO ; Jia-ju LÜ ; Zheng LIU ; Xiao-lei FANG ; Xun-bo JIN ; Ming-zhen YUAN
Chinese Medical Journal 2009;122(8):921-926
BACKGROUNDSeveral isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma (RCC) at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms' activity in RCC.
METHODSThe specimens of tumours (T) and clinically normal tissues (N) adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by chi(2) test or Fisher's exact test.
RESULTSp53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the Delta133p53beta isoform, which was not detected in the normal tissue. Of the six isoforms, p53beta mRNA was significantly overexpressed in tumour samples (P < 0.001), and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53gamma (19T/22N), Delta133p53 (33T/26N), Delta133p53beta (2T/0N), and Delta133p53gamma (13T/9N). Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings.
CONCLUSIONSAlthough all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53beta might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Protein Isoforms ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult
5.Analysis of adverse events following 2009 influenza A (H1N1) vaccinoprophylaxis in Beijing.
Liang MIAO ; Li LU ; Jiang WU ; Luo-dan SUO ; Dong-lei LIU ; Mei-ping SUN ; Xing-huo PANG ; Ying DENG ; Xiao-li WANG
Chinese Journal of Preventive Medicine 2010;44(10):884-887
OBJECTIVETo evaluate the safety of 2009 influenza A (H1N1) vaccine based on mass immunization initiative in Beijing.
METHODThere were 2 113 280 people were vaccinated during September to December 2009. The information of adverse events following immunization (AEFI) was collected through surveillance system, and descriptive methodology was used for data analysis.
RESULTSA Total of 612 AEFI cases were reported, among which there were 321 vaccine reaction cases following immunization, 203 coincidental illness cases, 82 psychogenic reaction cases, and 6 pending cases. The rates of rare reactions and common reactions associated with vaccination were respectively 5.54/100 000 (117/2 113 280) and 9.65/100 000 (204/2 113 280). The rate of serious rare reaction was 0.19/100 000 (4/2 113 280). The rates of vaccine reactions in urban, suburb and county were 16.87/100 000 (36/213 519), 17.81/100 000 (187/1 049 817) and 11.53/100 000 (98/849 944), respectively. The rates of rare reaction and common reaction in different age groups were between 3.65/100 000 (6/164 604) to 8.99/100 000 (27/300 176), and between 0.61/100 000 (1/164 604) to 22.06/100 000 (85/385 275). The 117 rear vaccine reaction cases were mainly allergic reaction (107 cases), and the 204 common vaccine reaction cases were mainly fever (176 cases). There were 91.90% (295/321) vaccine reactions occurred within 24 hours of administration, and all cases had improved consequence.
CONCLUSIONThe mostly symptoms of AEFI cases during the period of 2009 influenza A (H1N1) vaccinoprophylaxis were anaphylaxis and fever. The types of adverse reactions and the level of serious events are consistent with the anticipation. There were no rear or new events occurred.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Fever ; chemically induced ; Humans ; Hypersensitivity ; epidemiology ; Immunization ; Influenza A Virus, H1N1 Subtype ; Influenza Vaccines ; adverse effects ; Influenza, Human ; epidemiology ; prevention & control ; Male ; Mass Vaccination ; Middle Aged ; Population Surveillance ; Young Adult
6.An epidemiological study on HIV sexual transmission in married spouse in Yunnan province
Jin-Lei QI ; Hong-Bing LUO ; Yan-Ling MA ; Xiao-Jing AN ; Yan-Ling YANG ; Jun-Li HUO ; Hui GUO ; Lin LU ; Man-Hong JIA
Chinese Journal of Epidemiology 2012;33(2):173-176
Objective To understand the status of HIV sexual transmission among HIV- sero-discordant spouses and HIV-sero-accordant spouses in Yunnan province,to discuss the related factors and to provide evidence for HIV prevention and control strategy.Methods Five places with serious epidemic and 3 moderate ones were voluntarily,randomly selected.According to time sequence,300 spouses (600 people) with stable marriage were interviewed with questionnaire.Results HIV-sero-accordant spouses occupied for 40.7% of the total spouses under survey,with the others were HIV-sero-discordant ones.Among the ones that had already been diagnosed in the families,sexual transmission was their main mode of transmission,which was accounted for 68.3%,followd by IDU as 19.7%.After disclosed the HIV test outcomes to their spouses,63.4% HIV-sero-discordant spouses and 47.0% HIV-sero-accordant ones changed their sexual behaviors.The rates of consistent condom use among the HIV-sero-discordant spouses increased from 16.8% to 95.0%,and in HIV-sero-accordant spouses increased from 8.2% to 60.9%.Data were analyzed by multi-factor logistic regression.Factors on influencing the sexual transmission in spouses would include condom use,frequency of sexual contacts and sexual transmission disease (STD) status etc.Conclusion The main transmission mode for the first HIV infected spouse was sexual transmission.Factors influencing sexual transmission in spouses would include condom use,frequency of sexual contacts,STD situation and husband was the first one being infected in the families,etc.Disclosure of the HIV results to the spouses could make a significant changes in the frequencies of sexual contact as well as the rate of condom use.
7.Comparative study on sepsis models induced by Escherichia coli subtypes
Jing-Jing ZHANG ; Xian-Bin KONG ; Jing-Rui HUO ; Lei WANG ; Ying LIU ; Xiao-Hui YANG ; Yi TIAN ; Zhen-Jiang HOU ; Feng CHEN ; Xu-Yi CHEN ; Shi-Zhong SUN ; Tian-Guang XIA ; Zhong-Lei SUN ; Meng-Qiang HUANG ; Ying-Fu LIU
Tianjin Medical Journal 2018;46(6):585-589
Objective To investigate the degrees of injury severity of sepsis models made by different kinds of Escherichia coli. Methods The 152 mice were randomly divided into control group, DH5α group, 44102 group, and 25922 group, with 38 rats in each group. DH5α group, 44102 group and 25922 group were intraperitoneally injected with 300 μL of Escherichia coli DH5α, 44102 and 25922 at the concentration of 1.0 × 109CFU/kg to prepare sepsis models of different kinds of Escherichia coli. Control group was injected intraperitoneally with the same amount of normal saline. (1) After 8 h, four mice were taken from each group for peripheral blood bacterial culture . (2) After 12 h, ten mice in each group were used for measuring serum levels of TNF-α and IL-6 by enzyme-linked immunosorbent assay (ELISA). (3) Western blot assay was used to determine the serum levels of high-mobility group protein (HMGB1) in four mice of each group. (4) Ten mice in each group were used to measure serum levels of alanine transaminase (ALT), aspartate aminotransferase (AST), creatinine (CR) and blood urea nitrogen (BUN) by automatic biochemical analyzer. (5) After liver, lung and kidney tissues were fixed with formaldehyde, hematoxylin-eosin (HE) staining was performed (n=10 for each group). Results In DH5α group, 44102 group and 25922 group, bacteria, inflammatory cytokines TNF-α, IL-6 and HMGB1 protein, liver and kidney indicators ALT, AST, CR and BUN showed a sequential increasing trend (P<0.01). The severe degrees of alveolar structure damage, hepatic cell infiltration and renal glomerular atrophy were DH5α group, 44102 group and 25922 group in turn. There were no obvious damages of lung, liver or kidney tissues in control group. Conclusion Escherichia coli 25922 induces severe sepsis injury and can be used to study the animal models of the initial inflammatory phase of sepsis. Escherichia coli 44102 induces moderate damage of sepsis and can be used in animal models that do not require definitive sepsis staging experiments. Escherichia coli DH5α induces less damage of sepsis and can be used to explore immunosuppressive therapy of the animal model of sepsis.
8.Study on the risk factors related to severe acute respiratory syndrome among close contactors in Beijing.
Xing-Huo PANG ; Dong-Lei LIU ; Xiao-Hong GONG ; Fu-Jie XU ; Ze-Jun LIU ; Zhen ZHANG ; Xu-Hong WANG ; Xin-Ran JIAO ; Ling-Yan SU ; Gui-Hua GAO
Chinese Journal of Epidemiology 2004;25(8):674-676
OBJECTIVETo understand the risk factors on severe acute respiratory syndrome (SARS) among their contacts and to develop effective strategy for its control.
METHODSAvailable epidemiological data of SARS cases and close contacts were reviewed and analyzed by SPSS.
RESULTSOut of the 2195 close contacts, 138 (6.3%) were diagnosed as SARS. Among colleagues and classmates of SARS patients, the infection rate was 0.36% versus 31.71% in contacts among families and hospitals, 0.77% in schools. No one was infected among 459 close contacts to SARS in the working unit.
CONCLUSIONSAmong close contacts, factors that facilitating transmission would include: time, extent, frequency and place of contact to the patients, as well as factors related to close contacts as way, time of isolation and age. One of the epidemiological characteristics was that SARS were as clustered in the family among those close contacts. It is important to control the spread of SARS through supervision on the close contacts to patients.
Adult ; Aged ; China ; epidemiology ; Contact Tracing ; Cross Infection ; transmission ; Family Health ; Female ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; Male ; Middle Aged ; Patient Isolation ; Quarantine ; statistics & numerical data ; Retrospective Studies ; Risk Factors ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; transmission
9.Study of immunogenicity after primary vaccination by different sequential program of inactivated poliovirus vaccine and oral poliovirus vaccine.
Li LU ; Xiao-mei LI ; Dong-lei LIU ; He-run ZHANG ; Zhu-jia-zi ZHANG ; Hai-hong WANG ; Fang LIU ; Zhao-qi NING ; Li-wen ZHANG ; Ping CHU ; Yan-tao XIE ; Ying XU ; Juan LI ; Xing-huo PANG ; Ying DENG
Chinese Journal of Preventive Medicine 2012;46(6):510-513
OBJECTIVETo evaluate immunogenicity after primary vaccination by different sequential program of inactivated poliovirus vaccine (IPV) and oral poliovirus vaccine (OPV).
METHODSChildren of 2 months old (60-89 days) selected in Beijing were assigned to 4 groups, 1 dose IPV plus 2 doses OPV (I-O-O, 122 children), 2 doses IPV plus 1 dose OPV(I-I-O, 103 children), 3 doses IPV (I-I-I, 114 children), and 3 doses OPV (O-O-O, 106 children), and were vaccinated at the age of 2, 3, 4 months. Polio neutralizing antibody titers against poliovirus types 1, 2, and 3 were tested and protective rates were calculated before the 1st dose, after the last dose, and after the 1st and 2nd dose of IPV.
RESULTSAfter the primary immunization, geometric mean titers (GMT) of polio neutralizing antibody titers against poliovirus types 1, 2, and 3 were 788.32, 738.42 and 631.17 in O-O-O group, 212.02, 262.30 and 537.52 in I-I-I group, 940.35, 929.72 and 940.35 in I-O-O group and 901.09, 1102.68 and 1110.12 in I-I-O group (F values were 47.71, 53.84, and 9.81 respectively, all P values<0.01). The protective rate of three types among each group was 98.1% (104/106)-100.0% and the difference was not statistically significant (P>0.05). After the 1(st) dose of IPV, the GMT were 18.88, 37.77, 24.64 and the protective rate was 82.6% (122/138)-96.4% (133/138); after the 2nd dose of IPV, GMT were 177.03, 168.25, 321.86 and the protective rate was 99.1% (108/109)-100.0% (109/109) in antibody types 1, 2 and 3, respectively.
CONCLUSIONGMT of polio neutralizing antibody titers against poliovirus is higher after vaccination by sequential program of IPV and OPV than that by IPV or OPV 3-doses program. High level of protective rate after 2 doses of IPV in I-I-O group may lead to better protection from vaccine associated paralytic poliomyelitis (VAPP). Sequential program of IPV and OPV can be used to maintain high level of herd immunity and to prevent VAPP, and the I-I-O sequential program should be the first choice.
Humans ; Immunization Schedule ; Infant ; Poliovirus Vaccine, Inactivated ; administration & dosage ; immunology ; Poliovirus Vaccine, Oral ; administration & dosage ; immunology ; Vaccines, Attenuated ; immunology
10.Genetic analysis of 21 fetuses with high suspicion of congenital skeletal malformation by prenatal ultrasound
Ke YANG ; Yuwei ZHANG ; Guiyu LOU ; Na QI ; Bing KANG ; Hai XIAO ; Dong WU ; Yuan GAO ; Xingxing LEI ; Fengyang WANG ; Xiaodong HUO ; Bing ZHANG ; Shixiu LIAO
Chinese Journal of Perinatal Medicine 2022;25(1):28-34
Objective:To explore the genetic etiology of fetuses with high suspicion of congenital skeletal malformation detected by prenatal ultrasound.Methods:This retrospective study collected 21 pregnant women with highly suspected fetal skeletal malformation indicated by ultrasound (the couples had no skeletal malformation) at Institute of Medical Genetics, Henan Provincial People's Hospital from January 2019 to August 2020. Amniotic fluid/umbilical cord blood of the fetus and peripheral blood of the couples were obtained for karyotype analysis, chromosomal microarray analysis, and whole-exome sequencing. Sanger sequencing was performed for the "pathogenic" "suspected pathogenic" "variants of uncertain significance" variants detected by whole exome sequencing. Genetic etiology of the 21 fetuses was described.Results:A total of five chromosomal abnormalities were detected, including four cases of trisomy 21 and one trisomy 18. Chromosome microarray analysis detected one case of abnormal copy number variation, 16 p11.2 microdeletion syndrome. Ten cases of monogenic diseases were found by whole exome sequencing and eight genes were involved ( SGMS2, FGFR3, DYNC2H1, WDR35, TBX5, COL2A1, FGFR2, and ALPL). Totally, 14 variations were detected, among which seven were novel variations (c.8129T>A, c.7126G>A, c.10307_10320del, and c.2641G>T in DYNC2H1 gene; c.3085G>A and c.491G>A in WDR35 gene; c.1070G>T in COL2A1 gene). Conclusions:For fetus, whose parents have no skeletal malformation, highly suspected of congenital malformation of skeletal system by prenatal ultrasound, genetic factor is the primary reason, including chromosomal abnormalities, copy number variations, and monogenic mutations.