1.The co-vaccination effects of BCG and Poly I:C on the development of T cell subsets in neonatal BALB/c mice
Yun HE ; Enmei LIU ; Xiqiang YANG ; Chonghai LIU ; Xin LI
Chinese Journal of Microbiology and Immunology 2008;28(2):130-133
Objective To explore the effects of BCG and Poly I:C co-vaccination on the development of spleen T cell subsets of neonatal BALB/c mice. Methods Neonatal BALB/c mice were inoculated with BCG and/or Poly I:C intraperitoneally within 2-3 d after birth. Four weeks later, spleen cells of mice were isolated and the percentage of CD3+ CD8+ IFN-γ+,CD3+ CD8-IFN-γ+,CD3+ CD8+ IL-4+,CD3+ CD8- IL-4+,CD4+ Foxp3+ T cells,which represent Tc1,TH1,Tc2,TH2,Treg cells,respectively,were tested by flow cytometry at single cell level,and the ratios of TH 1/TH 2 and Tc1/Tc2 were calculated. Results The percentages of TH1 and Tc1 cells of BCG-vaccinated mice,Poly I:C-vaccinated mice and BCG plus Poly I:C-vaccinated mice were significantly higher than that of control mice(P<0.05 or P<0.01),and there was no difference among the three vaccinated group. The ratios of TH1/TH2 and total IFN-γ/IL-4 of the three vaccinated groups were higher than that of control group,but not the ratio of Tc1/Tc2. The TH1/TH2 ratio of BCG plus Poly I:C-vaccinated group was higher than that of BCG-vaccinated group(P<0.05).The percentages of Trge cells showed no difference among the four groups(P>0.05). Conclusion BCG and Poly I:C co-vaccination can significantly increase the number of Tc1 and TH 1 cells and TH 1/TH2 ratio in spleen cells. BCG and Poly I:C vaccination may have a synergistic effect on TH 1/TH2 ratio of spleen cells in neonataI mice. The percentage of CD4+ Foxp3+ T cells among four groups showed no significant difference.
2.Expression of EphA2 and E-cadherin in colorectal carcinoma
Chonghai DONG ; Ghunxia GUO ; Ling QU ; Jie LI ; Dexian WANG
Journal of Chinese Physician 2009;11(1):64-66
Objective To explore the expression and clinical significance of EphA2 and E-eadherin in colorectal carcinoma.Methods The expression of EphA2 and E-eadherin in 67 cases of eoloreetal carcinoma and 28 cases of normal large intestine tissues were determined by immunohistochemieal method(S-P method),and their relationship to elinicopathological characteristics were analyzed.Results The positive expression of EphA2 and negative expression of E-cadherin in colorectul carcinoma tissues were significantly higher than those in normal intestine tissues(P<0.01).F111e positive expression of EphA2 and negative expression of E-eadherin were positively correlated with tumor histological grade,invasive depth and lymph node metastasis(P<0.01 orP<0.05),but not correlated with tumor~ross type(P> O.05).The expression of EphA2 was negatively related with E-cadherin.Conclusions The abnormal expression of EphA2 and E-cadherin may be together involved in the development and progression of eolorectal carcinoma,It may be a good marker for monitoring the malignant degree and the prognosis of colorectal carcinoma by combining E-eadherin and EphA2.
3.Role of miR-486-5p in apoptosis of human bone marrow mesenchymal stem cells induced by hydrogen peroxide
Ming HU ; Jiao LI ; Ningning LIU ; Zhenjun HUANG ; Chonghai WU ; Yun ZHONG ; Shiming LIU
Chinese Journal of Pathophysiology 2015;(3):524-529
AIM:To investigate the role of microRNA-486-5p (miR-486-5p) in the apoptosis of human bone marrow mesenchymal stem cells (hMSCs) induced by hydrogen peroxide (H2O2).METHODS: The hMSCs were cul-tured in vitro and exposed to serum-free medium and H2O2(10 mmol/L).The changes of miR-486-5p expression in oxida-tive stress-related apoptosis of hMSCs were measured by real-time PCR.The hMSCs were transfected with miR-486-5p mimic or inhibitor at concentration of 30 nmol/L by Lipofectamine RNAiMAX.The effect of miR-486-5p on H2 O2-induced decrease in cell viability was evaluated by MTT assay.Hoechst 33342 staining and flow cytometry were applied to determine the role of miR-486-5p in the apoptosis of hMSCs.The protein expression was evaluated by Western blotting.Caspase-3 ac-tivity was determined using a caspase-3 activity kit.RESULTS:Compared with control group, the expression of miR-486-5p significantly decreased after treated with H2O2(P<0.05).In addition, over-expression of miR-486-5p in the hMSCs reduced the cell viability, accelerated apoptosis, down-regulated Bcl-2/Bax ratio, caspase-3 enzyme precursor content and phosphorylation of Akt, and activated caspase-3 activity.Conversely, down-regulation of miR-486-5p significantly inhibited H2 O2-induced cell apoptosis and the caspase-3 activity, increased cell viability and up-regulated Bcl-2/Bax ratio and phos-phorylation level of Akt.CONCLUSION:Over-expression of miR-486-5p promotes H2 O2-induced hMSCs apoptosis, and repression of miR-486-5p protects hMSCs from H2 O2-induced cellular apoptosis, which may be mediated by regulating Akt signaling pathway.
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. Running status of Qinghai provincial hand, foot and mouth disease laboratory network in 2017
Lixia FAN ; Zhuoma BA ; Chonghai LI ; Shuangying JIANG ; Shengcang ZHAO ; Dongmei YAN ; Tianjiao JI
Chinese Journal of Experimental and Clinical Virology 2018;32(3):286-288
Objective:
To evaluate the running status of Qinghai provincial hand foot and mouth disease (HFMD) laboratory network in 2017.
Methods:
The surveillance database developed in Qinghai provincial HFMD laboratory network in 2017 were analyzed, and the indicators for the running status of HFMD laboratory network in Qinghai province were evaluated.
Results:
It was shown that 574 samples of suspected HFMD cases were detected by real time RT-PCR in 2017, and 368 were positive, the positive rate was 64.11%. Then 121 virus strains were isolated, the results of sequencing and analysis showed that 100 strains were EV71 with C4a genotype, 16 strains were CA16 with B1b genotype, and 5 strains belonged to the other enterovirus. In addition, Qinghai provincial HFMD network labs passed all the confirmatory test and proficiency test, and on-site review held by national HFMD laboratory in 2017, respectively.
Conclusions
Qinghai provincial HFMD laboratory network has been established and running well. It provided important scientific basis for HFMD surveillance in Qinghai province.
6. Genetic characteristics of VP1 region of enterovirus Coxsackievirus A16 in Xining city from 2017 to 2018
Haijie GENG ; Liping REN ; Yongjian FENG ; Huirong ZHANG ; Xiaohong WANG ; Chonghai LI ; Xiaotong WANG ; Lixia FAN
Chinese Journal of Experimental and Clinical Virology 2019;33(5):482-484
Objective:
To investigate the genetic characteristics of VP1 coding region of enterovirus Coxsackievirus A16(CV-A16) and etiological features of hand, foot and mouth disease (HFMD) in 2017 in Xining city.
Methods:
The pharyngeal swab specimens were collected from HFMD patients, and detected by real-time reverse transcription-polymerase chain reaction (RT-PCR). For CV-A16 positive samples, virus isolation was performed. Then RNA was extracted, and then VP1 coding region was amplified by RT-PCR. The phylogenetic tree was constructed by comparing with other genotypes and sub-genotypes strains of EV-A71.
Results:
It was shown that 70 strains of CV-A16 were isolated from 2017 to 2018 in Xining city. In 2017, 10 strains were isolated and divided into two different lineages by phylogenetic analysis, 3 strains of B1a and 7 stains of B1b. In 2018, 60 stains were isolated, which were all belong to B1b.
Conclusions
B1a and B1b of CV-A16 are prevalent in Xining city from 2017 to 2018, in which B1b is the prominent isolates.
7. The operational status of Qinghai provincial measles laboratory network in 2012—2017
Lixia FAN ; Haotao TANG ; Zhuoma BA ; Shengcang ZHAO ; Chonghai LI ; Shuangying JIANG ; Zhen ZHU ; Huiling WANG
Chinese Journal of Experimental and Clinical Virology 2018;32(2):166-168
Objective:
To evaluate the running status of Qinghai provincial measles laboratory network during 2012—2017.
Methods:
To analyze serological and virological surveillance database developed in Qinghai provincial measles laboratory network in 2012—2017, and evaluate the indicators for the running status of measles laboratory network in Qinghai Province.
Results:
It was shown that 5 763 suspected measles cases were reported in Qinghai Province during 2012—2017, and 4 167 serum samples were collected from suspected measles cases, the collection rate is 72.31%; among them, 3 697 were IgM positive for measles and the positive rate was 88.72%; 68 were IgM positive for rubella and the positive rate was 1.63%. And 515 throat swab specimens were collected from suspected measles cases, 82 measles virus isolates were obtained and the positive rate was 15.92%. The result of sequencing and analysis showed that all the measles viruses belonged to genotype H1 and subgenotype H1a, which were predominant genotype circulated in China in recent years. In addition, Qinghai provincial measles network labs passed all the serological confirmatory test and proficiency test, and on-site review held by national or provincial measles laboratory and WHO during 2012—2017, respectively.
Conclusions
Qinghai provincial measles laboratory network has been established and running well. It provided important scientific basis for measles elimination in Qinghai province.
8. Genetic characteristics of human enterovirus A type 71 in Qinghai province during 2016-2017
Lixia FAN ; Weijun WANG ; Zhuoma BA ; Shengcang ZHAO ; Chonghai LI ; Shuangying JIANG ; Dongmei YAN ; Tianjiao JI
Chinese Journal of Experimental and Clinical Virology 2018;32(5):501-504
Objective:
To investigate the genetic characteristics of enterovirus A 71 (EV-A71) and etiological features of hand, foot and mouth disease (HFMD) in Qinghai province from 2016 to 2017.
Methods:
Specimens were collected from HFMD patients in Qinghai province, and detected by real-time reverse transcription-polymerase chain reaction (RT-PCR). For EV-A71 positive samples, virus was isolated and RNA was extracted, and then VP1 coding region was amplified by RT-PCR. The phylogenetic tree was constructed by comparing with other genotypes and sub-genotypes strains of EV-A71.
Results:
It was shown that 114 strains of EV-A71 were isolated in Qinghai province from 2016 to 2017, which all belonged to genotype C4a, and could be divided to two different lineages by phylogenetic analysis. From 2016 to 2017, the epidemic strains of EV-A71 in the different transmission chains of Qinghai province was closely related to other provinces of China.
Conclusions
C4a was the dominant genotype of EV-A71 in Qinghai province from 2016 to 2017, and no other genotype was detected. In addition, EV-A71 isolated from Qinghai province co-evolved with EV-A71 in other provinces of China.