1.dalbinol induces apoptosis of human colon cancer cells through ROS/Dvl/GSK-3β/β-catenin pathway
Feilong LI ; Xin WU ; Hongbo LIAO ; Shuangli QIU ; Xiaohui ZHU ; Liao CUI ; Hua WU
Chinese Pharmacological Bulletin 2016;32(12):1694-1698
Aim To investigate the effects of dalbinol on proliferation and apoptosis of human colon cancer HCT1 16 cells and its mechanisms.Methods Anti-proliferative effect of dalbinol was evaluated by MTT assay.The morphological changes of apoptosis were observed by Hoechst33342 staining.Apoptotic rate and ROS generation were analyzed by flow cytometry.The related proteins of Wnt/β-catenin pathway and the ap-optosis-associated proteins expression were measured by Western blot.Results The growth of HCT1 16 treated with dalbinol was inhibited in a dose and time dependent manner with IC50 (4.8 ±0.53 ),(2.5 ± 0.43)and (0.6 ±0.22)μmol·L-1 at 24,48 and 72 h,respectively.Typical morphological changes of ap-optosis such as cell shrinkage,karyopyknosis and nu-clear condensation were observed by Hoechst33342 staining.Meanwhile,the apoptotic rate and intracellu-lar ROS generation of dalbinol were both increased dose-dependently. Western blot results showed that dalbinol could activate the expression of cleaved Caspase-3 and cleaved PARP by decreasing anti-apop-totic protein levels such as Bcl-2 and Mcl-1 and in-creasing pro-apoptotic protein levels such as Bax and Bim,which induced further apoptosis.Moreover,dal-binol can reduce the protein expression of the total and nuclear β-catenin,but not cytoplasmic β-catenin by suppressing the protein expression of Dvl-2 and GSK-3β(pS9 ),as well as its target proteins c-Myc and Sur-vivin.Conclusion dalbinol can induce apoptosis in colon cancer HCT1 16 cells by upregulating the intra-cellular ROS generation and suppressing Dvl/GSK-3β/β-catenin pathway.
2.Genetic Characteristics of Type 2 Vaccine-derived Poliovirus in Shanxi Province (China) in 2014.
Dongrei YAN ; Xiaolei LI ; Yong ZHANG ; Jianfang YANG ; Shuangli ZHU ; Dongyan WANG ; Chuangye ZHANG ; Hui ZHU ; Wenbo XU
Chinese Journal of Virology 2015;31(2):157-163
The World Health Organization redefined the type 2 vaccine-derived poliovirus (VDPV) in 2010. To study the genetic characteristics and evolution of type 2 VDPV under this new definition, we conducted genome sequencing and analyses of type 2 VDPVs isolated from one patient with acute flaccid paralysis in Shanxi province (China) in 2014. Nucleotide sequencing revealed that the full-length of type 2 VDPV is 7439 bases encoding 2207 amino acids with no insertion or deletion of nucleotides compared with Sabin2. One nucleotide substitution identified as a key determinant of the attenuated phenotype of the Sabin 2 strain (A-G reversion at nucleotide nt 481 in the 5-end of the untranslated region) had reverted in the Shanxi type 2 VDPV. The other known key determinant of the attenuated phenotype of the Sabin 2 strain (U-->C reversion at nt2909 in the VP1 coding region that caused a Ile143Thr substitution in VP1) had not reverted in the Shanxi VDPV. The Shanxi type 2 VDPV was S2/S1 recombinant, the crossover site of which mapped to the 3-end of the 3D region (between nt 6247 and nt 6281). A phylogentic tree based on the VP1 coding region showed that evolution of the Shanxi type 2 VDPV was independent of other type 2 VDPVs detected worldwide. We estimated that the strain circulated for approximately = 11 months in the population according to the known evolution rate. The present study confirmed that the Chinese Polio Laboratory Network could discover the VDPV promptly and that it played an important part in maintenance of a polio-free China.
Amino Acid Sequence
;
Base Sequence
;
Capsid Proteins
;
chemistry
;
genetics
;
China
;
Humans
;
Infant
;
Male
;
Molecular Sequence Data
;
Phylogeny
;
Poliomyelitis
;
virology
;
Poliovirus
;
chemistry
;
genetics
;
metabolism
;
Poliovirus Vaccines
;
adverse effects
;
chemistry
;
genetics
;
metabolism
;
Sequence Alignment
3.Genetic Characteristics of Coxsackievirus Group A Type 4 Isolated from Patients with Acute Flaccid Paralysis in Shaanxi, China.
Dongyan WANG ; Yi XU ; Yong ZHANG ; Shuangli ZHU ; Yuan SI ; Dongmei YAN ; Hui ZHU ; Qian YANG ; Tianjiao JI ; Wenbo XU
Chinese Journal of Virology 2016;32(2):145-149
We analyzed the genetic characteristics of coxsackievirus A4 (CV-A4) based on the entire VP1 coding region. Samples were isolated from patients with acute flaccid paralysis (AFP) in Shaanxi, China from 2006 to 2010. We wished to ascertain the predominant genotype and the relationship between CV-A4 infection and AFP. Sixty-eight non-polio enteroviruses were inoculated onto RD cells (to increase the virus titer) and molecular typing was undertaken. The entire VP1 coding region was amplified. Percentage of CV-A4 was 10.3% (7/68). Analyses of genetic identify and creation of phylogenetic trees revealed that CV-A4 could be classified into A, B and C genotypes. Seven CV-A4 strains from Shaanxi and other CV-A4 strains from China formed an independent evolution lineage located in group 4 and belonged to the C2 sub-genotype. These data suggested that CV-A4 strains of sub-genotype C2 were the predominant genotypes in China. These strains co-evolved and co-circulated with those from other provinces in China, so continued monitoring of CV-A4 (by clinical and genetic surveillance) should be enhanced.
China
;
Enterovirus A, Human
;
classification
;
genetics
;
isolation & purification
;
Enterovirus Infections
;
virology
;
Genotype
;
Humans
;
Paralysis
;
virology
;
Phylogeny
;
Viral Proteins
;
genetics
4. Virological monitoring and analysis of polioviruses in Qinghai 2017
Lixia FAN ; Zhuoma BA ; Shengcang ZHAO ; Chonghai LI ; Shuangli ZHU
Chinese Journal of Experimental and Clinical Virology 2019;33(1):58-60
Objective:
To evaluate 2017 poliovirus surveillance in Qinghai Province.
Methods:
According to the World Health Organization (WHO) 4 th edition of the polio laboratory manual procedure for virus isolation, the isolated L20B positive strain was identified as intratypic differentiation (ITD) by the China Center for Disease Control and Prevention, CDC). The National Polio Laboratory performed the nucleotide sequence determination of the capsid protein VP1 coding region of poliovirus (PV) and analyzed the poliovirus surveillance and the result of analysis of the cases with acute flaccid paralysis (AFP) reported in Qinghai Province in 2017 and stool samples of healthy children.
Results:
In 2017, Qinghai CDC Polio Laboratory received specimens of 211 AFP cases and healthy stool samples. PV2 strains were isolated with a separation rate of 0.95%. Non-polio enterovirus (NPEV) strains were isolated from 25 strains with the isolation rate of 11.85%. Two PVs were used for ITD. All of them were vaccine-associated strains.
Conclusions
In 2017, the Qinghai CDC Polio Laboratory did not find any poliovirus and vaccine-derived poliovirus in the AFP cases and stool samples from healthy persons, maintaining the polio-free status.
5.A genome sequence of novel SARS-CoV isolates: the genotype, GD-Ins29, leads to a hypothesis of viral transmission in South China.
E'de QIN ; Xionglei HE ; Wei TIAN ; Yong LIU ; Wei LI ; Jie WEN ; Jingqiang WANG ; Baochang FAN ; Qingfa WU ; Guohui CHANG ; Wuchun CAO ; Zuyuan XU ; Ruifu YANG ; Jing WANG ; Man YU ; Yan LI ; Jing XU ; Bingyin SI ; Yongwu HU ; Wenming PENG ; Lin TANG ; Tao JIANG ; Jianping SHI ; Jia JI ; Yu ZHANG ; Jia YE ; Cui'e WANG ; Yujun HAN ; Jun ZHOU ; Yajun DENG ; Xiaoyu LI ; Jianfei HU ; Caiping WANG ; Chunxia YAN ; Qingrun ZHANG ; Jingyue BAO ; Guoqing LI ; Weijun CHEN ; Lin FANG ; Changfeng LI ; Meng LEI ; Dawei LI ; Wei TONG ; Xiangjun TIAN ; Jin WANG ; Bo ZHANG ; Haiqing ZHANG ; Yilin ZHANG ; Hui ZHAO ; Xiaowei ZHANG ; Shuangli LI ; Xiaojie CHENG ; Xiuqing ZHANG ; Bin LIU ; Changqing ZENG ; Songgang LI ; Xuehai TAN ; Siqi LIU ; Wei DONG ; Jun WANG ; Gane Ka-Shu WONG ; Jun YU ; Jian WANG ; Qingyu ZHU ; Huanming YANG
Genomics, Proteomics & Bioinformatics 2003;1(2):101-107
We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence
;
China
;
Cluster Analysis
;
Gene Components
;
Genetic Variation
;
Genome, Viral
;
Genotype
;
Molecular Sequence Data
;
Phylogeny
;
Reverse Transcriptase Polymerase Chain Reaction
;
SARS Virus
;
genetics
;
Sequence Analysis, DNA
;
Severe Acute Respiratory Syndrome
;
genetics