1.Cloning and bioinformatics analysis of gene AsTP2 transactivated by arsenic trioxide with suppression subtractive hybridization
Shunhua WU ; Yujian ZHENG ; Yuexin ZHANG
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
Objective To study cloning and the primary function of a new gene AsTP2 transactivated by arsenic trioxide. Methods Using suppression subtractive hybridization (SSH) technique, the mRNA was isolated from HepG2 cells treated with arsenic trioxide (5?mol/L) and 0.9 percent sodium chloride, respectively, then cDNA was synthesized. SSH method was employed to analyze the differentially expressed DNA sequences between the two groups. From the subtractive cDNA library of genes transactivated by arsenic trioxide, the coding sequence of a new gene was obtained by bioinformatics method, and amplified by the method of reverse transcription polymerase chain reaction (RT-PCR). Results The novel gene was named as AsTP2, which was logged in the GenBank with the accession number AY744366. AsTP2 of 1119 nucleotides (nt), coding a protein of 372 amino acid residues (aa). Conclusion A new gene has been recognized as the new target transactivated by arsenic trioxide. The results will give a new clue to explore the molecular carcinogenic mechanism of inorganic arsenic.
3.Correlation of clinicopathological features and prognosis in patients with colorectal cribriform comedo-type adenocarcinoma
Lihua ZHANG ; Xiaoyan CHEN ; Chen WANG ; Shunhua CHEN ; Liyu CAO
Chinese Journal of Clinical and Experimental Pathology 2015;(7):776-779
Purpose To investigate the clinicopathological features and prognosis in patients with colorectal cribriform comedo-type ad-enocarcinoma. Methods Compared with 278 cases of colorectal common-type adenocarcinoma, 24 cases of colorectal cribriform come-do-type adenocarcinoma were analyzed for clinical stage, lymphovascular invasion, lymph node metastasis and genetic characteristics. Results The percentage of clinical Ⅲ and Ⅳ (87. 5%) and lymph node metastasis rate (87. 5%) in patients with colorectal cribri-form comedo-type adenocarcinoma was obviously higher than that in colorectal common-type adenocarcinoma (42. 4%,42. 4%) (P<0. 05). The percentage of lymphovascular invasion (87. 5%) of colorectal cribriform comedo-type adenocarcinoma were significantly higher than that of common-type adenocarcinoma (22. 7%) (P<0. 05). There were no statistical significance of age, sex, and molec-ular genetics of KRAS and mutated BRAF mutation between two groups ( P>0. 05 ) . Conclusions There is a higher clinical stage and poorer prognosis including unique morphological characteristics, highly lymphovascular invasion and lymph node metastasis in pa-tients with colorectal cribriform comedo-type adenocarcinoma.
4.Proteomics analysis of rat liver fibrosis caused by sodium arsenite
Shunhua WU ; Jing LI ; Lulu ZHANG ; Chenlu WANG ; Xiaoying LIU
Chinese Journal of Endemiology 2016;35(3):182-188
Objective To study the mechanism of liver fibrosis in rats caused by chronic exposure through drinking water containing sodium arsenite,to identify the differential proteins via proteomics technique.Methods Totally 40 healthy 8-week-old male Sprague Dawley (SD) rats of specific pathogen free (SPF) grade were randomly divided into 4 groups,which were control group (deionized water),0.68,1.36 and 2.73 mg/kg sodium arsenite (iAs3+) treated groups,respectively.The rats were fed with iAs-treated drinking water freely for 24 consecutive weeks.Twenty-four hour urine sample,blood and liver samples were collected.Hepatic fibrosis indices,specifically,type Ⅲ precollagen (PC Ⅲ),type Ⅳ collagen (Ⅳ-C),hyaluronic acid (HA) and laminin (LN) were detected by enzymelinked immunoassay (ELISA).Based on the isobaric tags for relative and absolute quantitation (iTRAQ) reagent 8-plex experiment,combined with 2DLC-MS/MS,the proteins in rats liver tissue of the medium dose group and the high dose group were compared with the those of control groups.Results ①The serum HA contents in the C (control) group,the L (low dose) group,the M (medium dose) group and the H (high dose) group were (198.51 ± 16.64),(218.39 ± 34.98),(261.72 ± 30.56) and (297.31 ± 35.72) ng/L;the serum PCⅢ contents in C,L,M and H groups were (15.32 ± 2.15),(16.78 ± 2.64),(19.51 ± 0.85) and (21.42 ± 1.63) μg/L;the serum LN contents in C,L,M and H groups were (734.57 ± 86.00),(792.65 ± 94.15),(916.83 ± 84.40) and (1 008.09 ± 64.17) μg/L;the serum Ⅳ-C contents in C,L,M and H groups were (52.34 ± 14.65),(59.72 ± 12.84),(74.38 ± 4.83) and (78.46 ± 4.30) μ.g/L,respectively.The differences in serological indices of liver fibrosis between-groups were statistically significant (F =21.136,19.957,22.007,14.288,all P < 0.05).In multiple comparison of serum HA,PCⅢ and LN,there were no statistical significant differences between L group and C group.M and H groups were higher than L group and C group,significant statistical difference was found between H group and M group (all P < 0.05).②Combining iTRAQ with 2DLC-MS/MS,based on the confidence threshold of protein (unused protScore) > 1.3 and at least 1 matched peptides within the 95% confidence interval,2 948 proteins were identified.Totally 2 162 proteins were detected in three groups compared with Venn diagram,after removing significant different proteins in C group,687 up-regulated proteins and 548 down-regulated proteins were identified in M group;633 up-regulated proteins and 519 downregulated were found in H group;the differences of protein expression between M and H groups were not statistically significant (P > 0.05).③Up-regulated proteins related to the metabolism including AS3MT,MAT,SHMT,CHDH,CTH,CSAD and BHMT in M and H groups;of the two kinds of proteins of MTR,METK1 was up-regulated and F1LRB8 was down-regulated.Proteins associated with GSH including Gsta1,Gsta4,Gsta5,Gstt1,Gstt2,Gstk1,Gstp1,Gstm1,Gstm2,Gstm3,Gss,Gpx1,Gpx4,Esd,Hagh,Glo1,Mgst1 and B6DYQ5 which were all up-regulated.Proteins associated with liver fibrosis were Hic-5,Gss and six kinds of Tpm,and six kinds of Tpm subunits including two kinds of Tpm1,three kinds of Tpm2 and one kind of Tpm3 which were all up-regulated.Conclusions There is liver accumulation of arsenic after chronic arsenic exposure and resulting in liver fibrosis and decline of liver function.Expressions of AS3MT,MTR,MAT,SHMT,BHMT,CHDH,CTH and CSAD are up-regulated;arsenic meta bolism methionine cycle,folic acid cycle and sulfur transfer pathways are closely related.GSH plays an important role in arsenic metabolism and liver fibrosis,Hic-5,GSS and TPM may be associated with the occurrence of liver fibrosis.
5.Activation of rat hepatic stellate cells and changes of DNA methyltransferase 1, 3a mRNA expression levels induced by sodium arsenite
Shunhua WU ; Lulu ZHANG ; Chenlu WANG ; Jing LI
Chinese Journal of Endemiology 2017;36(1):32-36
Objective To observe the activation of rat hepatic stellate cells (HSC-T6) and the changes of methyltransferase (DNMT)1,DNMT3a mRNA expression with different doses of sodium arsenite stimulation.Methods HSC-T6 cells were exposed to a final concentration of 0 (control),5 (low dose),15 (medium dose) and 25 (high dose) μmol/L sodium arsenite in culture medium for 24,48 and 72 h,cells and cell culture supernatant were harvested.Enzyme-linked immunosorbent assay (ELISA) kit was used to measure fibrosis factors contents of type Ⅰ collagen (COL-1),type 11Ⅲ collagen (COL-3) and alpha smooth muscle actin (α-SMA) and quantitative real-time PCR was used to detect the expression levels of DNMT1 and DNMT3a mRNA.Results Different arsenic exposure time (24,48,72 h) had a significant effect on COL-1,COL-3 and α-SMA contents in HSC-T6 cells (F =249.574,328.493,3 157.436,all P < 0.01);Different arsenic exposure content (low,medium,high dose groups) had a significant effect on COL-1,COL-3 and α-SMA contents in HSC-T6 cells (F =3 946.521,1 006.399,13 025.770,all P < 0.01).After arsenic exposure for 24 and 48 h,the expression levels of DNMT1 mRNA in high dose group (4.33 ± 0.24,2.34 ± 0.43) were higher than those of control group (1.00 ± 0.00,1.00 ± 0.00,all P < 0.05).At the same arsenic exposure levels (low,medium or high dose),the expression level of DNMT1 mRNA was declined with prolongation of sodium arsenite stimulation time (all P < 0.05).After arsenic exposure for 48 and 72 h,the expression levels of DNMT3a mRNA in high dose group (2.23 ± 0.50,5.02 ± 0.23) were higher than those of control group (1.00 ± 0.00,1.00 ± 0.00,all P < 0.05).The expression levels of DNMT3a mRNA in medium and high dose groups at 72 h (3.80 ± 0.14,5.02 ± 0.23) were higher than those of 24 h (3.03 ± 0.12,0.42 ± 0.15,all P < 0.05).Conclusion HSC-T6 cells are obviously activated with pro-fibrotic effect;the expression levels of DNMT1 and DNMT3a mRNA are both up regulated in HSC-T6 cells after being exposed to sodium arsenite.
6.Expression of miR-21 in prostate cancer and its clinical significance
Cong ZHANG ; Liyu CAO ; Yu YIN ; Shunhua CHEN ; Yujie LI
Chinese Journal of Clinical and Experimental Pathology 2016;32(12):1365-1367
Purpose To investigate the expression and clinical significance of microRNA-21 (miR-21) in patients with prostate cancer (PCa).Methods The human tissue specimens were paraffin-embedded blocks which were collected from the Department of Pathology.miR-21 was detected by in situ hybridization (ISH) in 90 cases of PCa and 69 cases of benign prostatic hyperplasia (BPH).Results The ISH data showed that the positive rate of miR-21 in PCa (71.1%) was significantly higher than that in BPH (24.6%) (P < 0.05),the expression of miR-21 correlated with the Gleason grade (P < 0.05),and no relationship was observed with other clinicpathological features (P > 0.05).Conclusion The miR-21 plays an important role in the pathogenesis of PCa,which can be a new diagnostic target for PCa.
7.Comparison of somatotypes on 29 ethnic groups in China
Lianbin ZHENG ; Shunhua LU ; Bo DING ; Xinghua ZHANG
Acta Anatomica Sinica 2012;43(1):130-134
Objective To analyze the characteristics of somatotypes of South Asian type, North Asian type and East Asian type, and to compare the main differences among them. Methods The characteristics of somatotypes among South Asian type, North Asian type and East Asian type in 29 Chinese Mongolian ethnic groups were compared by using the Heath-Carter anthropometric somatotype. The reasons for differences of South Asian type were that North Asian type were analyzed by using principal component analysis. Results The male groups of North Asian type were endomorphic mesomorph and the South Asian types were balanced mesomorph, while the East Asian types differed greatly from each other.The female groups of North Asian types and the East Asian types were mesomorphic endomorph and the South Asian types were endomorphic mesomorph. The somatotypes of East Asian types were similar to North Asian types but were greatly different from South Asian types.Principal components analysis showed that the main differences between South and North of male groups lied first in ectomorphy and then in endomorphy.The differences between female groups of South and North were mainly on endomorphy. Conclusion The ethnic group of North Asian type is higher than South Asian type in endomorphy but lower in ectomorphy.
8.Different expression patterns of β-catenin and its correlation with clinicopathological facters in colorectal cancer
Wen JIN ; Shunhua CHEN ; Yu YIN ; Cong ZHANG ; Liyu CAO
Chinese Journal of Clinical and Experimental Pathology 2017;33(6):596-600,605
To observe different expression patterns of β-catenin and its clinical significance in colorectal cancer (CRC).Methods A total of 181 cases of CRC tissues and 30 cases of normal colorectal tissue were investigated by immunohistochemistry for the expression of β-catenin.Results The expression rate of β-catenin was 56.9% (103/181) in CRC,and higher than that in normal colorectal tissue (P < 0.05).The overexpression of nuclear β-catenin was significantly correlated with histological differentiation,lymph node metastasis and Dukes' stage in CRC (P < 0.05),and no relationship with other pathological parameters,such as age,gender and the depth of infiltration.The incomplete membranous expression of β-catenin was significantly correlated with histological differentiation,the depth of infiltration,lymph node metastasis and Dukes' stage in CRC (P < 0.05).The high expression of nuclear β-catenin related to histological differentiation and Dukes' stage in CRC (P < 0.05).In the follow-up data of 82 cases of CRC,the expression of nuclear β-catenin was associated with poor prognosis,and the 5-year survival rate was significantly lower than that of self-control groups (P < 0.05).Conclusion β-catenin plays important roles in colorectal carcinogenesis.Abnormal expression of β-catenin was related to the aggressive progression of CRC and may be helpful for evaluating the prognosis of patients with CRC.β-catenin is expected to become a new target for diagnosis and treatment of CRC in future.
9.Effects of chronic exposure to sodium arsenite on liver damage in rats
Guanxin DING ; Jia HUANG ; Qin LIN ; Rongxian XIA ; Shunhua WU ; Ling ZHANG
Journal of Preventive Medicine 2022;34(9):887-892
Objective:
To examine the effect of chronic exposure to sodium arsenite on liver damages in rats.
Methods:
Fifty-six healthy adult SD rats (28 males and 28 females) were randomly divided into 4 groups. Rats in the low-, medium- and high-dose groups were given sodium arsenite solutions at doses of 2, 10 and 50 mg/L for successive 24 weeks, while animals in the control group were given deionized water. The rat body and liver weights were measured and the liver coefficient was estimated. The urine arsenic level was detected using atomic fluorescence spectrometry, and hepatic tissue sections were stained with uranium acetate and lead citrate for morphological observations under an electron microscope.
Results:
The body weights of both male and female rats appeared a tendency towards a rise with the duration of exposure to sodium arsenite (male rat: Wald χ2=3 610.621, P<0.001; female rat: Wald χ2=2 186.217, P<0.001, and there were no significant differences in the rat body weight 24 weeks post-exposure to sodium arsenite in each group, while there was an interaction between time and group (male rat: Wald χ2=15.874, P=0.001; Wald χ2=9.460, P=0.024). There were significant differences in the rat liver weight and liver coefficient in each group (male rat: F=18.964 and 29.968, both P<0.001; female rat: F=11.919 and 15.070, both P<0.001), with the lowest liver weight (10.17±1.15) g and liver coefficient (1.99±0.21)% measured in male rats in the high-dose group, and the highest liver weight (12.91±1.29) g and liver coefficient (4.10±0.56)% in female rats in the high-dose group. The median urine arsenic levels (interquartile range) were 25.60 (30.27), 146.56 (101.06), 1 034.68 (600.06) and 3 796.98 (19 966.89) μg/L in rats in the control, low-dose, medium-dose and high-dose groups, respectively (χ2=50.211, P<0.001), and the urine arsenic level was significantly higher in the medium- and high-dose groups than in the control group (both P<0.001). Hepatic edema was seen in rats in the low- and medium-dose groups, and hepatic edema, focal hepatic cell necrosis, hyperplasia of bile capillaries and peri-bile capillary endolysis were observed in rats in the high-dose group.
Conclusions
Chronic exposure to arsenic may cause morphological alterations of rat hepatic tissues, and the rat hepatic damage aggravates with the dose of exposure to arsenic.
10.Selective Attentional Networks Deficits in Parents of Children With Autism Spectrum Disorder: Evidence From Attention Network Test
Tian QIN ; Zihan WANG ; Xiaoming LI ; Shunhua ZHANG
Psychiatry Investigation 2023;20(8):721-729
Objective:
We primarily aimed to investigate the attention network function among parents of children with autism spectrum disorder (ASD) using the Attention Network Test (ANT). The secondary objective was to observe whether the three attention networks of all participants were related to each other.
Methods:
We included 28 parents of children with ASD and 28 well-matched parents of typically developing children. All participants underwent the neuropsychological assessment and ANT test. The three distinct attention networks, including alerting, orienting, and executive control, were also measured.
Results:
Compared with controls, parents of children with ASD showed less-efficient alerting and executive control network (all p<0.05), but not orienting network (p=0.74). No significant correlation was found between the alerting, orienting, and executive control network for either group.
Conclusion
Our findings showed that parents of children with ASD had deficits in alerting and executive control attention functions. The deficits are indications of a broad autism phenotype.