1.Screening for pathogenesis-related genes of osteosarcoma using gene microarray
Guo-Dong LI ; Zheng-Dong CAI ; Yin-Quan ZHANG ; Ming RU ; Fang JI ;
Chinese Journal of Cancer Biotherapy 2006;0(05):-
Objective:To screen for the pathogenesis-related genes of osteosarcoma and to assess their roles for the de- velopment of osteosareoma.Methods:Total RNA was extracted from 3 ATCC osteosarcoma cell lines and an osteoblastic cell line and was used to synthesize biotinylated cRNAs;the latter were hybridized to Affymetrix~(?)GeneChip~(?)U133A ar- rays and a gene with more than 2 folds of change was selected.Ten of the differentially expressed genes were chosen and the primers were designed and the synthesized.Then SYBR~(?)Green real-time PCR(RT-PCR)method was used to detect the expression of the 10 genes in 9 fresh osteosarcoma specimens.ABI Prism 7 000 system was used to analyze the differ- ent expression between osteosarcoma cell line and osteoblastic cell line.Results:We identified 58 up-regulated and 142 down-regulated genes in the 3 osteosareoma cell lines.Many of the genes were firstly reported to be related to the patho- genesis of osteosarcoma.These differentially expressed genes were mainly involved in energy and material metabolism,on- cogene,signal transduction gene,transcription- related genes,cell cycle-related genes,cell apoptosis-related gene,im- mune response gene,tumor suppressor genes,etc.The array results of 10 randomly selected genes were further verified by the RT-PCR in 9 fresh osteosarcoma specimens.Conclusion:Many genes are involved in the pathogenesis of osteosarcoma. Gene microarray can help to discover the genes related to the pathogenesis of osteosarcoma,which may lay a foundation for studying the molecular mechanism of osteosarcom.
2.Application of chromosomal microarray analysis for fetuses with talipes equinovarus
Qiaoli GUO ; Fang FU ; Ru LI ; Xiangyi JING ; Tingying LEI ; Jin HAN ; Xin YANG ; Li ZHEN ; Min PAN ; Can LIAO
Chinese Journal of Obstetrics and Gynecology 2016;51(7):484-490
Objective To investigate the application of fetuses with talipes equinovarus (TE) using chromosomal microarray analysis (CMA) technology. Methods From May 2012 to June 2015, 54 fetuses were found with TE and with or without other structural anomalies by prenatal ultrasound. Karyotyping was taking for them all, and the fetuses with normal karyotypes took another CMA test. The data were analyzed with CHAS software. Finally all the cases were followed up to know about their pregnancy outcomes. Results One of the 54 cases was detected with abnormal karyotype which was trisomy 18 (2%, 1/54). CMA was undertaken to the remaining fetuses, they were divided into 2 groups, including isolated TE group (n=38) and complex TE group (n=15). The detection rate of clinical significant copy number variations (CNV) by CMA was 11% (6/53), while isolated and complex TE group were 5% (2/38) and 4/15, respectively (P=0.047). Of the 53 cases, 51 cases were successfully followed up. Eleven cases were found without TE after birth, and the false positive rate (FPR) of TE was 22%(11/51). Conclusions Whole-genome high-resolution CMA increased the detection rate by 11% in fetuses with TE. With the FPR and the detection rate of the clinical significant CNV of 2 groups, whole-genome CMA could be recommended to the fetuses with complex TE group but normal karyotypes. A series of ultrasonic tests should be suggested to the isolate TE group, while with the abnormal ultrasound, fetuses would be suggested to have CMA test for decreasing the rates of invasive prenatal diagnosis and FPR.
3.Tonal audiometry of GJB2 235delC single heterozygous mutation carriers.
Qi LI ; Ru-ping FANG ; Guo-jian WANG ; Fang LIU ; Pu DAI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(7):543-546
OBJECTIVETo analyze GJB2 235delC monoallelic mutation carrier individuals and test the possible presence and incidence of audiometric abnormalities among 30-60 years old carriers of the 235delC mutations.
METHODSA total of 32 unrelated subjects with nonsyndromic hearing loss were screened for the 235delC mutation. Tonal audiometric analysis was performed on the 235delC mutation carrier group and on a non-carrier control group.
RESULTSAudiometric evaluations in the control group showed the presence of thresholds within normal limits at all frequencies, while carriers of the 235delC mutation presented with decreased hearing at 1000 Hz and 2000 Hz (age 40-49 years and 50-59 years), and 4000 and 8000 Hz (age 30-59 years), P < 0.05. The hearing loss of carriers gradually increased with age.
CONCLUSIONSGJB2 235delC heterozygous carriers may be a risk group for high-frequency hearing loss. Hearing thresholds may deteriorate in the intermediate frequencies over the age of 40.
Adult ; Aged ; Audiometry, Pure-Tone ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Female ; Genotype ; Hearing Loss ; genetics ; physiopathology ; Heterozygote ; Humans ; Male ; Middle Aged ; Mutation
4.Effect of allergic rhinitis on disease condition and treatment in patients with juvenile-onset systemic lupus erythematosus.
Yan-Qing ZHOU ; Lan-Fang CAO ; Ru-Ru GUO ; Le-Ping LI ; Hui-Fen FANG
Chinese Journal of Contemporary Pediatrics 2017;19(5):510-513
OBJECTIVETo investigate the effect of allergic rhinitis (AR) and its intervention on disease condition and medications in patients with juvenile-onset systemic lupus erythematosus (JSLE).
METHODSThe clinical data of 96 children diagnosed with JSLE were collected, and according to the presence or absence of AR or other allergic diseases, they were divided into AR group (n=44), non-AR group (n=20), and non-allergic group (n=32). The children in the AR group were randomly administered with or without intervention (n=22 each). All the children were given standard JSLE treatment. The systemic lupus erythematosus disease active index (SLEDAI) and application of hormones and immunosuppressants were compared between groups.
RESULTSThe AR and non-AR groups had significantly higher SLEDAI scores, daily cumulative doses of glucocorticoids, and number of types of immunosuppressants used than the non-allergic group before treatment (P<0.05), while there were no significant differences between the AR and non-AR groups (P>0.05). After one month of treatment, the AR group with intervention had significantly lower SLEDAI scores and daily cumulative doses of glucocorticoids than the AR group without intervention (P<0.05), while there was no significant difference in the application of immunosuppressants between these two groups (P>0.05). After 3 and 6 months of treatment, the AR group with intervention had significantly lower SLEDAI scores, daily cumulative doses of glucocorticoids, and number of types of immunosuppressants than the AR group without intervention (P<0.05).
CONCLUSIONSJSLE combined with allergic diseases such as AR has an adverse effect on disease condition and treatment, and the intervention for AR helps with the control of JSLE.
Adolescent ; Child ; Child, Preschool ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Immunosuppressive Agents ; therapeutic use ; Interleukin-17 ; blood ; Interleukins ; Lupus Erythematosus, Systemic ; drug therapy ; immunology ; Male ; Rhinitis, Allergic ; complications ; Severity of Illness Index
5.Clinical significance of TGF-β1 in children with primary IgA nephropathy.
Hong-Tao ZHU ; Liang RU ; Yan-Fang GUO
Chinese Journal of Contemporary Pediatrics 2014;16(7):749-753
OBJECTIVETo investigate the clinical significance of transforming growth factor-beta 1 (TGF-β1) in children with primary IgA nephropathy (IgAN).
METHODSThirty children who were diagnosed with primary IgAN by renal biopsy between May 2008 and October 2012 were included in the study. Thirty healthy children were used as the control group. Urinary and blood TGF-β1 levels were measured using enzyme-linked immunosorbent assay, and the protein expression of TGF-β1 in the renal tissue was measured by immunohistochemistry. The correlation between TGF-β1 levels in blood, urine, and renal tissue and their relationship with clinical indices were analyzed.
RESULTSChildren with primary IgAN had significantly higher urinary and blood TGF-β1 levels than the control group (P<0.01). Urinary TGF-β1 level was positively correlated with the pathological grade of renal tissue (r=0.557, P=0.001), and a significant positive correlation was also found between the TGF-β1 expression in the renal tissue and the pathological grade of renal tissue (r=0.682, P<0.01). There was no correlation between TGF-β1 levels in blood and renal tissue (r=0.038, P=0.844).
CONCLUSIONSUrinary TGF-β1 level is significantly positively correlated with the pathological severity of disease in children with primary IgAN. Clinical measurement of urinary TGF-β1 may be of great practical value in predicting the progression and prognosis of chronic nephropathy.
Adolescent ; Child ; Female ; Glomerulonephritis, IGA ; metabolism ; pathology ; Humans ; Kidney ; chemistry ; pathology ; Male ; Transforming Growth Factor beta1 ; analysis ; physiology ; urine
6.Dynamic evaluation on body weight gain in premature infants and its significance.
Xi-fang RU ; Qi FENG ; Ying WANG ; Xin ZHANG ; Xing LI ; Jing-wen MENG ; Zai-chen GUO
Chinese Journal of Pediatrics 2010;48(9):661-667
OBJECTIVESTo investigate the incidence of intra- and extrauterine growth retardation (EUGR) and growth restriction in premature infants, and to illustrate the growth pattern of them in postnatal and infantile period.
METHODSAll premature infants were admitted to our neonatal intensive care unit (NICU) during the recent 7 years. The criteria for enrollment were (1) gestational age < 37 weeks; (2) single fetus; (3) admitted within the first 24 hours of life; (4) hospitalization period ≥ 14 days; (5) clinical follow-up persisted till ≥ 3 months of corrected gestational age. Intrauterine growth restriction (IUGR), EUGR and growth restriction were defined as having a measured growth value (weight) that was ≤ 10(th) percentile of Chinese infants' growth curve in corrected age on admission, discharge and follow-up period. Results were analyzed by using SPSS 12.0 statistical software package by chi-square test, rank-sum test, and t test.
RESULTSTwo hundred and thirty nine infants were involved, 134 were boys and 105 girls. The incidence of IUGR and EUGR assessed by weight was 25.5% and 40.6%, respectively. The lower the birth weight was, the higher the incidence of IUGR and EUGR was. The percentile of body weight in the growth curve at discharge was lower than that at birth (Z = -7.784, P = 0.000). The incidence of growth restriction assessed by weight was 20.5%, 15.0%, 8.8%, 17.0%, 10.4%, 10.1%, 11.9%, 7.0% at corrected gestational age of 38 - 40 weeks, corrected age of 28 d, 61 d, 91 d, 122 d, 152 d, 183 d, and 274 d, respectively. The incidences of growth restriction were stable when the corrected age was older than 91 days. The incidence of growth restriction in female premature infants at 183 days' corrected age was higher than that in male children (χ(2) = 6.181, P = 0.017), the incidence was 19.3% and 3.8% respectively. During the follow-up period, most of the average body weight of premature infants whose gestational age was < 32 weeks or birth weight ≤ 1500 g were lower than the 50(th) percentile of the growth curve except the average body weight of boys whose gestational age < 32 weeks at corrected age of 2 and 4 months.
CONCLUSIONSPremature and/or low birth weight infants are at high risk of growth restriction, especially very low birth weight infants. The incidence of growth restriction decreased with growth. Long-term prognosis requires further investigation.
Body Weight ; Female ; Fetal Growth Retardation ; Follow-Up Studies ; Humans ; Infant, Newborn ; Infant, Premature ; growth & development ; Male ; Weight Gain
7.Transforming growth factor-beta1 and Smad4 signaling pathway down-regulates renal extracellular matrix degradation in diabetic rats.
Qin YANG ; Ru-jia XIE ; Ting YANG ; Li FANG ; Bing HAN ; Guo-zhong ZHANG ; Ming-liang CHENG
Chinese Medical Sciences Journal 2007;22(4):243-249
OBJECTIVETo investigate the role of transforming growth factor-beta1 (TGF-beta1)/Smad4 pathway in development of renal fibrosis in streptozotocin (STZ)-induced diabetic nephropathy (DN) rats and explore its possible mechanism.
METHODSMale Wistar rats weighing 180-220 g were divided into 5 groups: group A (normal control), group B [diabetes mellitus (DM) 2 weeks], group C (DM 4 weeks), group D (DM 8 weeks), and group E (DM 16 weeks). Except for the normal control group, other groups were induced DM by single injection of STZ (55 mg/kg) respectively. Blood glucose level, serum creatinine, and 24-hour urine protein were examined. Expressions of TGF-beta1 and Smad4 protein and mRNA in kidney were detected using immunohistochemical technique, Western blot, and real-time PCR. mRNA expressions of stromelysin-1 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1), and collagen In in kidney were also detected by real-time PCR.
RESULTSThe levels of blood glucose, serum creatinine, and 24-hour urine protein in rats of group B, C, D, and E were higher than those of the control group. With the progression of renal fibrosis, the expressions of TGF-beta1 and Smad4 protein and mRNA in kidney of diabetic rats elevated. In addition, the renal MMP-3 mRNA expression diminished in diabetic rats, while TIMP-1 and collagen III mRNA increased.
CONCLUSIONSIn STZ-induced diabetic rats, the TGF-beta1/Smad4 appears to play an important role in renal fibrosis of DN. The increased expression of TGF-beta1 and Smad4 might result in the transcriptional regulation of downstream target genes of TGF-beta1/Smad4 pathway, which contributes to the progression of renal fibrosis in diabetic rats.
Animals ; Base Sequence ; DNA Primers ; Diabetes Mellitus, Experimental ; metabolism ; Down-Regulation ; Extracellular Matrix ; metabolism ; Kidney ; metabolism ; Male ; Rats ; Rats, Wistar ; Signal Transduction ; Smad4 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism
8.Mechanisms of human telomerase reverse transcriptase RNAi which increases hepatocellular carcinoma cell apoptosis induced by TRAIL.
Ru-Gang ZHANG ; Dian-Chun FANG ; Xiao-Yan NING ; Guo-An WANG
Chinese Journal of Hepatology 2007;15(1):32-36
OBJECTIVESTo investigate the mechanisms for human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) in increasing hepatocellular carcinoma cell apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL).
METHODSCell apoptosis was identified by flow cytometry analysis after annexin V/PI double staining. Expression of apoptosis-related proteins, procaspase-8, -9, -3, Bax, Bcl-2 and hTERT, were identified by Western blotting analysis; telomerase activity and telomere length were detected by telomeric repeat amplification protocol (TRAP) and telomere amount and length assay (TALA) methods.
RESULTSHepatocellular carcinoma cell apoptosis induced by TRAIL were all significantly increased by hTERT RNAi (P less than 0.05). For example, apoptosis rates were enhanced from 5.53% (untransformed) to 10.35% (transformed) in HepG 2 cells and from 14.73% to 77.24% in SMMC 7721 cells after being treated by 100 ng/ml TRAIL for 24 h. Moreover, activation of procaspase-8, -9 and -3 in transformed cells after being treated by TRAIL were all significantly raised (P less than 0.05) in a dose-dependent manner. The expression of procaspase-8, -9 and Bcl-2 were effectively augmented (P less than 0.05), but expressions of Bax and hTERT were strikingly decreased (P less than 0.05). Meanwhile, telomerase activity was apparently suppressed and telomere length was markedly shortened (P less than 0.05). There were no remarkable differences in these effects between control cells and the untransformed cells (P more than 0.05).
CONCLUSIONEnhanced cell apoptosis induced by TRAIL through hTERT RNAi may be related to up-regulation of procaspase-8 and -9 expressions. However the down-regulation of hTERT expression, reduced telomerase activity and shortened telomere length may not be related to expressions of Bcl-2 and Bax.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Telomerase ; genetics ; metabolism
9.Effects of hTERT RNAi on apoptosis of hepatocellular carcinoma cells induced by TRAIL.
Ru-gang ZHANG ; Dian-chun FANG ; Yuan-hui LUO ; Li-ping GUO
Chinese Journal of Hepatology 2006;14(6):435-440
OBJECTIVETo study the effects of human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) on biological characteristics of hepatocellular carcinoma cell lines HepG2 and SMMC-7721 and on apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).
METHODSSmall hairpin hTERT (shTERT) sequence was identified by PCR method; hTERT expressions, morphological features, cell proliferation and replicative senescence were respectively determined using RT-PCR, hematoxylin-eosin (HE) staining, growth curve and beta-galactosidase (b-Gal) staining; cell cycle and apoptosis were identified using flow cytometry after propidium iodide (PI) staining and annexin V/PI double staining.
RESULTSshRNA were found in 6/8 HepG2 and 6/6 SMMC-7721 cell clones transformed by the recombined plasmid pSilencer 3.1-H1 neo-shTERT. The interference rates of hTERT on HepG2 and SMMC-7721 were 100% and 43.3% respectively. Cells in G2-M phases increased from 7.1% to 10.6% and from 6.9% to 7.9% respectively; and the percentage of replicative senenscence cells increased from 0 to 20.4% and from 3.6% to 10.0% respectively. The nucleus/cytoplasm ratios of the cells were obviously decreased after hTERT RNAi treatment. Moreover, apoptosis of hepatocellular carcinoma cells and apoptosis induced by TRAIL were strikingly increased by hTERT RNAi (P < 0.05). For example, apoptosis rates were increased from 3.5% to 5.2% in HepG2 cells and from 4.8% to 7.9% in SMMC-7721 cells after hTERT RNAi treatment. Apoptosis rates were increased from 5.3% to 10.4% in HepG 2 cells and from 13.9% to 77.2% in SMMC-7721 cells after being treated by 100 ng/ml TRAIL for 24 h. However, there were no remarkable changes between control cells and untransformed cells.
CONCLUSIONhTERT RNAi not only has a significant effect on biological characteristics of hepatocellular carcinoma cells, but also obviously can increase cell apoptosis induced by TRAIL.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Telomerase ; genetics ; Tumor Cells, Cultured
10.Early growth of preterm infants with prolonged hospitalisation.
Xi-Fang RU ; Qi FENG ; Ying WANG ; Xin ZHANG ; Xing LI ; Jing-Wen MENG ; Zai-Chen GUO
Singapore medical journal 2012;53(12):832-839
INTRODUCTIONThis study aimed to determine the early growth patterns of preterm infants who required prolonged hospitalisation in terms of body weight Z-score, and to explore the influencing factors and predictors of their growth.
METHODSThe criteria of enrolment included preterm birth, singleton pregnancy, hospitalisation within the first 24 hours of life, hospital stay ≥ 28 days and clinical follow-up beyond 91 days of corrected age. Body weight Z-scores and the incidence of underweight infants were reviewed periodically, and the influencing factors and possible predictors of growth analysed.
RESULTSBody weight Z-scores of all infants of gestational age (GA) groups kept decreasing, with a trough seen at 36 weeks corrected gestational age (CGA). At corrected full-term, body weight Z-scores for all birth weight groups achieved birth level and were higher than that at 36 weeks CGA. Body weight Z-scores at 61 days corrected age was (-0.300 × GA [weeks] + 0.210 × birth weight [g] + 0.682 × body weight Z-score) at 40 weeks CGA. The cut-off values for body weight Z-score at birth (cut-off, -1.79; sensitivity, 100%; specificity, 91.3%) and 61 days corrected age (cut-off, -1.95; sensitivity, 100%; specificity, 97.1%) were selected to predict the risk of being underweight at 183 days corrected age.
CONCLUSIONEarly growth restriction is a practical problem in preterm infants with prolonged hospitalisation. Body weight Z-scores at 40 weeks CGA and 61 days corrected age can be used to predict body weight gain prior to 183 days corrected age in these infants.
Female ; Follow-Up Studies ; Gestational Age ; Growth Disorders ; epidemiology ; etiology ; Humans ; Incidence ; Infant, Newborn ; Infant, Premature ; growth & development ; Infant, Premature, Diseases ; epidemiology ; etiology ; Length of Stay ; trends ; Male ; Pregnancy ; Retrospective Studies ; Singapore ; epidemiology