1.Isolation,identification and genetic analysis of a three-lineage NADC30 PRRSV recombinant strain
Bingyan CAI ; Yangyang QIAO ; Tianxin LIU ; Weiyong ZUO ; Yongjuan WANG ; Kai ZHANG ; Hui LU ; Haiming WANG
Chinese Journal of Veterinary Science 2025;45(3):420-426
In June 2018,sows at a pig farm in Jiamusi,Heilongjiang Province,suffered a large num-ber of miscarriages,and subsequently weaned piglets at the farm began to show persistent high fe-ver symptoms at around 35 days of age,with some pigs having a fever of more than 41.5 ℃.In or-der to determine the cause of this outbreak,63 clinical samples from this farm were tested.The re-sults showed that 60 out of 63 samples were positive for the porcine reproductive and respiratory syndrome virus(PRRSV)antigen.Subsequently,PRRSV antigen-positive plasmid was transfected into African green monkey embryonic kidney cells(Marc-145),and after three generations of blind transmission,indirect immunofluorescence assay(IFA)was performed.The results showed that one PRRSV strain,named HLJ38,was successfully isolated.Then the whole genome of HLJ38 strain was sequenced and then analyzed in detail by bioinformatics software.Sequence analysis showed that there were three deletions of 131 amino acids(323-433 aa,483 aa and 504-522 aa)in the derived sequence of Nsp2 gene of HLJ38 strain,which was consistent with the molecular ge-netic marker of NADC30 PRRSV.The phylogenetic tree analysis showed that HLJ38 and NADC30 PRRSV in GenBank belong to lineage 1 subgroup,and the nucleotide homology of HLJ38 and NADC30 PRRSV in GenBank was only 85.2%and 84.6%.Recombinant analysis showed that HLJ38 was a recombinant NADC30-like PRRSV,and the recombinant gene fragments were de-rived from multiple strains,among which the fragment of 1-201 nt was provided by VR2332 strain and fragment of 6 641-8 061 nt derived from the HP-PRRSV strain.In summary,the re-sults showed that the outbreak in this pig farm may be caused by the recombination of PRRSV strains among different lineages,and the recombinant circulating strains not only have certain pathogenicity but also suggest that the existing commercial vaccines provide limited cross-protec-tion against them.Recombination between different lineages increases the genetic diversity of PRRSV and aggravates the difficulty of prevention and control of PRRS in pig farms.Therefore,it is necessary to continuously monitor the epidemic dynamics of PRRSV in pig farms and take effec-tive measures in time to curb the spread of PRRS.
2.Study on the Relationship between the Changes of Four Indexes Related to Plasma Ferroptosis and the Prognosis after TACE in Patients with Hepatocellular Carcinoma
Fei YANG ; Jicheng GAO ; Song LIU ; Huixiao ZUO ; Weiyong GONG ; Zhe ZHANG ; Tao PENG
Journal of Modern Laboratory Medicine 2025;40(5):78-81,87
Objective To analyze the relationship between the expression of ferroptosis markers in the tumor microenvironment(TME)and the prognosis of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC).Methods This prospective observational study included 100 HCC patients who received TACE treatment at Langfang Hospital of Traditional Chinese Medicine from March 2019 to June 2021 as the study subjects.The levels of 8-isoprostaglandin F2α(8-iso-PGF2α),4-hydroxy-2-nonenal(4-HNE),8-hydroxy-2'deoxyguanosine(8-OH-dG)and hepcidin in plasma were evaluated by ELISA kit at baseline(1 day before TACE),1 day after TACE and 4~8 weeks.The changes of ferroptosis related markers during TACE treatment were compared.The difference between the level of 8-iso-PGF2α,4-HNE and the baseline 1 day after TACE treatment was recorded as △8-iso-PGF2α,△4-HNE.Results Compared with the baseline,the levels of 8-iso-PGF2α and 4-HNE increased significantly and the level of hepcidin decreased significantly one day after TACE treatment,and the differences were statistically significant(t=8.03,16.29,2.92,all P<0.05).Compared with 1 day after treatment,the levels of 8-iso-PGF2α,4-HNE decreased and the level of 8-OH-dG increased at 4~8 weeks after TACE treatment,and the differences were statistically significant(t=9.12,17.17,2.63,all P<0.05).Multivariate COX analysis showed that △8-iso-PGF2α,△4-HNE and 8-iso-PGF2α 1 day after TACE treatment were independent factors affecting the overall survival after TACE(Wald χ2=5.205,13.801,6.054,all P<0.05).The survival time of patients with △4-HNE>2.01 μg/ml was significantly longer than that of patients with △4-HNE≤2.01 μg/ml(Log-rank=5.718,P=0.017),and that of patients with△8-iso-PGF2α>1.75ng/ml was sig-nificantly longer than that of patients with△8-iso-PGF2≤1.75ng/ml(Log-rank=4.163,P=0.041).Conclusion The prognosis of HCC patients who are in a state of high ferroptosis(4-HNE and 8-iso-PGF2 increased)at 1 day after TACE treatment is better,which indicate that ferroptosis mediated HCC death induced by TACE treatment.
3.Isolation,identification and genetic analysis of a three-lineage NADC30 PRRSV recombinant strain
Bingyan CAI ; Yangyang QIAO ; Tianxin LIU ; Weiyong ZUO ; Yongjuan WANG ; Kai ZHANG ; Hui LU ; Haiming WANG
Chinese Journal of Veterinary Science 2025;45(3):420-426
In June 2018,sows at a pig farm in Jiamusi,Heilongjiang Province,suffered a large num-ber of miscarriages,and subsequently weaned piglets at the farm began to show persistent high fe-ver symptoms at around 35 days of age,with some pigs having a fever of more than 41.5 ℃.In or-der to determine the cause of this outbreak,63 clinical samples from this farm were tested.The re-sults showed that 60 out of 63 samples were positive for the porcine reproductive and respiratory syndrome virus(PRRSV)antigen.Subsequently,PRRSV antigen-positive plasmid was transfected into African green monkey embryonic kidney cells(Marc-145),and after three generations of blind transmission,indirect immunofluorescence assay(IFA)was performed.The results showed that one PRRSV strain,named HLJ38,was successfully isolated.Then the whole genome of HLJ38 strain was sequenced and then analyzed in detail by bioinformatics software.Sequence analysis showed that there were three deletions of 131 amino acids(323-433 aa,483 aa and 504-522 aa)in the derived sequence of Nsp2 gene of HLJ38 strain,which was consistent with the molecular ge-netic marker of NADC30 PRRSV.The phylogenetic tree analysis showed that HLJ38 and NADC30 PRRSV in GenBank belong to lineage 1 subgroup,and the nucleotide homology of HLJ38 and NADC30 PRRSV in GenBank was only 85.2%and 84.6%.Recombinant analysis showed that HLJ38 was a recombinant NADC30-like PRRSV,and the recombinant gene fragments were de-rived from multiple strains,among which the fragment of 1-201 nt was provided by VR2332 strain and fragment of 6 641-8 061 nt derived from the HP-PRRSV strain.In summary,the re-sults showed that the outbreak in this pig farm may be caused by the recombination of PRRSV strains among different lineages,and the recombinant circulating strains not only have certain pathogenicity but also suggest that the existing commercial vaccines provide limited cross-protec-tion against them.Recombination between different lineages increases the genetic diversity of PRRSV and aggravates the difficulty of prevention and control of PRRS in pig farms.Therefore,it is necessary to continuously monitor the epidemic dynamics of PRRSV in pig farms and take effec-tive measures in time to curb the spread of PRRS.
4.Study on the Relationship between the Changes of Four Indexes Related to Plasma Ferroptosis and the Prognosis after TACE in Patients with Hepatocellular Carcinoma
Fei YANG ; Jicheng GAO ; Song LIU ; Huixiao ZUO ; Weiyong GONG ; Zhe ZHANG ; Tao PENG
Journal of Modern Laboratory Medicine 2025;40(5):78-81,87
Objective To analyze the relationship between the expression of ferroptosis markers in the tumor microenvironment(TME)and the prognosis of transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC).Methods This prospective observational study included 100 HCC patients who received TACE treatment at Langfang Hospital of Traditional Chinese Medicine from March 2019 to June 2021 as the study subjects.The levels of 8-isoprostaglandin F2α(8-iso-PGF2α),4-hydroxy-2-nonenal(4-HNE),8-hydroxy-2'deoxyguanosine(8-OH-dG)and hepcidin in plasma were evaluated by ELISA kit at baseline(1 day before TACE),1 day after TACE and 4~8 weeks.The changes of ferroptosis related markers during TACE treatment were compared.The difference between the level of 8-iso-PGF2α,4-HNE and the baseline 1 day after TACE treatment was recorded as △8-iso-PGF2α,△4-HNE.Results Compared with the baseline,the levels of 8-iso-PGF2α and 4-HNE increased significantly and the level of hepcidin decreased significantly one day after TACE treatment,and the differences were statistically significant(t=8.03,16.29,2.92,all P<0.05).Compared with 1 day after treatment,the levels of 8-iso-PGF2α,4-HNE decreased and the level of 8-OH-dG increased at 4~8 weeks after TACE treatment,and the differences were statistically significant(t=9.12,17.17,2.63,all P<0.05).Multivariate COX analysis showed that △8-iso-PGF2α,△4-HNE and 8-iso-PGF2α 1 day after TACE treatment were independent factors affecting the overall survival after TACE(Wald χ2=5.205,13.801,6.054,all P<0.05).The survival time of patients with △4-HNE>2.01 μg/ml was significantly longer than that of patients with △4-HNE≤2.01 μg/ml(Log-rank=5.718,P=0.017),and that of patients with△8-iso-PGF2α>1.75ng/ml was sig-nificantly longer than that of patients with△8-iso-PGF2≤1.75ng/ml(Log-rank=4.163,P=0.041).Conclusion The prognosis of HCC patients who are in a state of high ferroptosis(4-HNE and 8-iso-PGF2 increased)at 1 day after TACE treatment is better,which indicate that ferroptosis mediated HCC death induced by TACE treatment.
5.Analysis of the correlation between estrogen level and retinopathy in postmenopausal patients with type 2 diabetes
Huan LIAO ; Meilan YANG ; Wei ZUO ; Wen YAN ; Weiyong ZHANG ; Jia CHENG ; Wenqing ZHONG
Clinical Medicine of China 2018;34(3):263-267
Objective To investigate the correlation between postmenopausal estrogen levels and diabetic retinopathy in patients with type 2 diabetes.Methods Two hundred cases of postmenopausal type 2 diabetes admitted to the Eighth Affiliated Hospital,Sun Yat-sen University from January 2015 to January 2017 were selected as the subjects and divided into the retinopathy group (93 cases) and the normal retina group (107 cases) according to the resuhs of fundus angiography.The general data of the two groups were compared,and the blood glucose index,blood lipid index and estrogen level were measured and compared.Logistic regression analysis was used to analyze the related factors of the patients with retinopathy.Results (1) The disease duration of patients in the retinopathy group was longer than that in the normal retina group ((14.67 ±3.48) years vs.(10.20±2.85) years;t=9.983,P<0.05).The fasting blood glucose (FBG),glycosylated hemoglobin of patients in the retinopathy group were higher than those in the normal retina group (FBG:(8.01 ± 1.21) mmol/L vs.(6.93 ± 1.20) mmol/L,FBG:(8.32 ± 1.56) mmol/L vs.(7.39 ± 1.43) mmol/L;t =6.324,4.397,P<0.05).The total cholesterol (TC),triglyceride (TG) and low density lipoprotein (LDL-C) of patients in the retinopathy group were higher than those in the normal retina group [TC:(4.93±0.87) mmol/ Lvs.(4.23± 0.69) mmol/L,TG:(1.91 ± 0.42) mmol/L vs.(1.63 ± 0.32) mmol/L,LDL-C:(2.61 ± 0.90) mmol/L vs.(1.78±0.53) mmol/L;t =6.340,5.340,8.067,P<0.05],while high density lipoprotein (HDL-C) was lower than that in the normal retina group ((1.23±0.37) mmol/L vs.(1.76±0.45) mmol/L;t =9.014,P <0.05).The E2 level in the retinopathy group was lower than that in the normal retina group ((23.91 + 8.51) ng/L vs.(32.18 + 9.80) ng/L;t=6.325,P<0.05).Logistic regression analysis showed that the duration of diabetes,FBG,HbAlc,TC,TG and LDL-C were risk factors of retinopathy (OR (95% CI) =1.049 (1.011 ~ 1.056),1.071(1.061 ~ 1.093),1.092(0.984~ 1.275),1.099(1.661 ~ 1.993),1.863(1.661 ~ 1.993),1.652 (1.461 ~ 1.893),P< 0.05),while HDL-C and E2 were protective factors of retinopathy (OR (95 % CI) =0.172(0.061 ~ 0.293),0.955 (0.861 ~ 1.093),P<0.0S).Conclusion Estrogen level is related to retinopathy in postmenopausal diabetic patients.It may be a protective factor for diabetic retinopathy.

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