Objective This study aimed to investigate the genetic diversity, differentiation, and population structure of Aedes albopictus across different geographical regions in Shandong Province, and to explore the relationship between genetic diversity and geographical distribution. Methods Between July and August 2024, ten Ae. albopictus populations were sampled from seven cities in Shandong Province. Genomic DNA was extracted from individual mosquitoes, and the mitochondrial cytochrome c oxidase subunit I(COⅠ)gene was amplified using PCR and sequenced. The obtained sequences were verified using BLAST and analyzed with MAFFT, MEGA 11, DnaSP v6.12, Arlequin 3.5, PopART 1.7, STRUCTURE 2.3.4 and the R packages adegenet and vegan to assess genetic diversity and population structure. Results A total of 229 COⅠ sequences(662 bp) was obtained, revealing ten variable sites with no insertions or deletions. The overall base composition showed an AT bias of 67.7%. Haplotype analysis identified 11 haplotypes, with Hap2 being the dominant and most widely distributed haplotype across all populations. Neutrality tests showed significant population expansion only in the Rizhao population. Mantel testing result revealed that geographical distance does not significantly impede gene flow. Overall, genetic differentiation among populations was low, indicating frequent gene flow. When combined with additional samples from Shanghai, Fujian, Guangdong, Yunnan, Hainan and Guangxi, STRUCTURE, UPGMA and DAPC analyses revealed two primary genetic clusters of Ae. albopictus. Conclusions Ae. albopictus populations in Shandong Province exhibit frequent gene flow, low genetic differentiation, and relatively low overall genetic diversity. However, the Rizhao population showed signs of recent expansion, highlighting the need for enhanced surveillance and targeted control measures in this area.

Objective To investigate the value of amide proton transfer(APT)imaging in evaluating lymph-vascular space inva-sion(LVSI)of cervical cancer.Methods A retrospective analysis was conducted on the data of cervical cancer patients with patho-logically confirmed LVSI status.Based on the presence of LVSI,the patients were divided into LVSI positive group and LVSI nega-tive group.All patients underwent diffusion weighted imaging(DWI)and APT imaging before treatment,and the apparent diffusion coefficient(ADC)and APT values of the lesions were measured.An independent sample t-test was used to compare the differences in ADC and APT values between the two groups.The sensitivity,specificity and area under the curve(AUC)of ADC,APT and ADC+APT in predicting LVSI were observed by receiver operating characteristic(ROC)curve,and the diagnostic performance of the three was compared by DeLong test.Results A total of 78 patients were included,of which 54 were LVSI negative and 24 were LVSI positive.The ADC values in the LVSI positive group were significantly lower than those in the LVSI negative group(P＜0.001),while the APT values in the LVSI positive group were significantly higher than those in the LVSI negative group(P＜0.001).The sensitivities of ADC,APT and ADC+APT in predicting LVSI were 79.17％,83.33％ and 87.50％,respectively,the specificities were 75.93％,55.56％ and 77.78％,respectively,and the AUC were 0.828,0.759 and 0.868,respectively,indicating that the diag-nostic performance of ADC+APT was better than that of ADC and APT alone(P＜0.001).Conclusion APT imaging can preop-eratively predict the presence of LVSI status in cervical cancer.When combined with ADC,its diagnostic accuracy is higher than that of APT alone,providing a new approach for evaluating LVSI in cervical cancer.

Objective To investigate the value of amide proton transfer(APT)imaging in evaluating lymph-vascular space inva-sion(LVSI)of cervical cancer.Methods A retrospective analysis was conducted on the data of cervical cancer patients with patho-logically confirmed LVSI status.Based on the presence of LVSI,the patients were divided into LVSI positive group and LVSI nega-tive group.All patients underwent diffusion weighted imaging(DWI)and APT imaging before treatment,and the apparent diffusion coefficient(ADC)and APT values of the lesions were measured.An independent sample t-test was used to compare the differences in ADC and APT values between the two groups.The sensitivity,specificity and area under the curve(AUC)of ADC,APT and ADC+APT in predicting LVSI were observed by receiver operating characteristic(ROC)curve,and the diagnostic performance of the three was compared by DeLong test.Results A total of 78 patients were included,of which 54 were LVSI negative and 24 were LVSI positive.The ADC values in the LVSI positive group were significantly lower than those in the LVSI negative group(P＜0.001),while the APT values in the LVSI positive group were significantly higher than those in the LVSI negative group(P＜0.001).The sensitivities of ADC,APT and ADC+APT in predicting LVSI were 79.17％,83.33％ and 87.50％,respectively,the specificities were 75.93％,55.56％ and 77.78％,respectively,and the AUC were 0.828,0.759 and 0.868,respectively,indicating that the diag-nostic performance of ADC+APT was better than that of ADC and APT alone(P＜0.001).Conclusion APT imaging can preop-eratively predict the presence of LVSI status in cervical cancer.When combined with ADC,its diagnostic accuracy is higher than that of APT alone,providing a new approach for evaluating LVSI in cervical cancer.

BACKGROUND:Ovarian tissue vitrification cryopreservation is one of the important methods for preserving fertility.Tanshinone ⅡA has various pharmacological activities,including anti-oxidation,inhibition of inflammatory response,and reduction of apoptosis,but its role as an additive for vitrification cryoprotection of ovarian tissue is still unclear.OBJECTIVE:To explore the protective effect of tanshinone ⅡA on vitrification cryopreservation of mouse ovarian tissue.METHODS:Twenty-five 6-week-old female KM mice were randomly selected and their ovarian tissues were randomly divided into five groups,with 10 ovaries per group.The fresh group was not cryopreserved.The frozen control group used vitrification cryoprotectant.The 0.5,2.5,and 5 μmol/L tanshinone ⅡA groups used vitrification cryoprotectant containing 0.5,2.5,and 5 μmol/L tanshinone ⅡA,respectively,and were cryopreserved in liquid nitrogen.After 3 days of storage,the cryopreserved tubes were taken out and thawed.The ovarian tissue and follicle morphology of each group were observed by hematoxylin-eosin staining,and the normal follicle morphology and survival rate were analyzed.The levels of superoxide dismutase,catalase,malondialdehyde,tumor necrosis factor-α,interleukin-1β,and interleukin-17 in the ovary were detected by enzyme-linked immunosorbent assay.RT-qPCR and western blot assay were used to detect the mRNA and protein expressions of nuclear factor E2-related factor 2(Nrf2)and heme oxygenase 1(HO-1)in the mouse ovary.RESULTS AND CONCLUSION:(1)Compared with the fresh group,the frozen control group had abnormal morphology of follicles at all levels in the ovary,decreased follicle survival rate(P＜0.05),decreased superoxide dismutase and catalase activities(P＜0.05);the levels of malondialdehyde,and tumor necrosis factor α,interleukin 1β,and interleukin 17 were all increased(P＜0.05),and the mRNA and protein expressions of Nrf2 and HO-1 were decreased(P＜0.05).(2)Compared with the frozen control group,different concentrations of tanshinone ⅡA could improve the morphology of follicles at all levels in the ovary,increase the survival rate of follicles,enhance the activities of superoxide dismutase and catalase,and reduce the levels of malondialdehyde,tumor necrosis factor α,interleukin 1β,and interleukin 17,increased the mRNA and protein expression of Nrf2 and HO-1 in a concentration-dependent manner,with 5 μmol/L tanshinone ⅡA having the most significant effect.(3)The results show that tanshinone ⅡA may reduce the oxidative stress level and inflammatory response of mouse ovarian tissue by mediating the Nrf2/HO-1 signaling pathway,thereby alleviating the reproductive damage caused by vitrification cryopreservation of mouse ovaries.

A quadruplex RT-qPCR method was developed for rapid identification and diagnosis of transmissible gastroenteritis virus(TGEV)of swine,porcine epidemic diarrhea virus(PEDV),porcine deltacorona virus(PDCoV),and porcine rotavirus type A(PoRVA).The full-length sequences of PEDV(77 strains),TGEV(63 strains),PDCoV(17 strains)that are prevalent in China,as well as the 85 VP6 gene sequences of PoRVA,were downloaded from the NCBI database for homology analysis.Based on the relatively conserved sequences,the corresponding primers and probes for each virus were designed and used to establish the quadruplex RT-qPCR method.After optimization of the probes and the reaction conditions,the specificity,sensitivity,and repeatability were determined.Using the established method,109 clinical samples of diarrhea were tested and further compared with the results by standard method.The results showed that the quadruplex RT-qPCR method established in this experiment has good amplification effect,with the C,value linearly correlated with the copies of templates(R2＞0.99).Specificity assay demonstrated that the quadruplex RT-qPCR method can identify TGEV,PEDV,PDCoV,PRoVA strains,and do not de-tect African swine fever virus(ASFV),porcine circovirus type 2(PCV2),porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV),pseudorabies virus(PRV)and other epidemic viruses.Sensitivity assay showed that the detection limits for TGEV,PEDV,PDCoV and PoRVA were 10,20,20 and 50 copies/μL,respectively.The method exhibits excellent reproducibility,with coefficients of variation(Cv)for both intra-and inter-assay repli-cates being less than 1％.Detection of 109 samples of diarrhea by this method yielded the coinci-dence rate of 100％with the industry standard,indicating high practical applicability.The devel-oped method possesses the advantages of strong strain compatibility,high sensitivity,strong speci-ficity,good repeatability and stability.It is suitable for virus diagnosis and large-scale clinical sam-ple testing,providing technical support for disease prevention and control as well as epidemiologi-cal investigation.

A quadruplex RT-qPCR method was developed for rapid identification and diagnosis of transmissible gastroenteritis virus(TGEV)of swine,porcine epidemic diarrhea virus(PEDV),porcine deltacorona virus(PDCoV),and porcine rotavirus type A(PoRVA).The full-length sequences of PEDV(77 strains),TGEV(63 strains),PDCoV(17 strains)that are prevalent in China,as well as the 85 VP6 gene sequences of PoRVA,were downloaded from the NCBI database for homology analysis.Based on the relatively conserved sequences,the corresponding primers and probes for each virus were designed and used to establish the quadruplex RT-qPCR method.After optimization of the probes and the reaction conditions,the specificity,sensitivity,and repeatability were determined.Using the established method,109 clinical samples of diarrhea were tested and further compared with the results by standard method.The results showed that the quadruplex RT-qPCR method established in this experiment has good amplification effect,with the C,value linearly correlated with the copies of templates(R2＞0.99).Specificity assay demonstrated that the quadruplex RT-qPCR method can identify TGEV,PEDV,PDCoV,PRoVA strains,and do not de-tect African swine fever virus(ASFV),porcine circovirus type 2(PCV2),porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV),pseudorabies virus(PRV)and other epidemic viruses.Sensitivity assay showed that the detection limits for TGEV,PEDV,PDCoV and PoRVA were 10,20,20 and 50 copies/μL,respectively.The method exhibits excellent reproducibility,with coefficients of variation(Cv)for both intra-and inter-assay repli-cates being less than 1％.Detection of 109 samples of diarrhea by this method yielded the coinci-dence rate of 100％with the industry standard,indicating high practical applicability.The devel-oped method possesses the advantages of strong strain compatibility,high sensitivity,strong speci-ficity,good repeatability and stability.It is suitable for virus diagnosis and large-scale clinical sam-ple testing,providing technical support for disease prevention and control as well as epidemiologi-cal investigation.

Aim To elucidate the underlying mecha-nism by which total triterpenoids extracted from Hove-nia dulcis(H-TP)enhance the sensitivity of A549/DDP cells to cisplatin.Methods The ARE-Nrf2 lu-ciferase reporter assay was applied to investigate the impact of H-TP on Nrf2 expression.Western blot was used to detect the protein levels of Keap-1/Nrf2/HO-1,Nrf2-GPX4 signaling pathway,apoptosis-related proteins of Bcl-2 and Bax.Further validation of its effects on Nrf2 was conducted by using Nrf2 activator/inhibitor.Results H-TP could enhance the sensitivi-ty of A549/DDP cells to cisplatin by modulating the expression of apoptosis-related proteins Bax and Bcl-2,inhibiting the Keap-1/Nrf2/HO-1/GPX4 signating pathway in A549/DDP cells,and inducing ferroptosis.Conclusion H-TP enhances the sensitivity of A549/DDP cells to cisplatin by inducing the Nrf2-mediated ferroptosis pathway.

Aim To elucidate the underlying mecha-nism by which total triterpenoids extracted from Hove-nia dulcis(H-TP)enhance the sensitivity of A549/DDP cells to cisplatin.Methods The ARE-Nrf2 lu-ciferase reporter assay was applied to investigate the impact of H-TP on Nrf2 expression.Western blot was used to detect the protein levels of Keap-1/Nrf2/HO-1,Nrf2-GPX4 signaling pathway,apoptosis-related proteins of Bcl-2 and Bax.Further validation of its effects on Nrf2 was conducted by using Nrf2 activator/inhibitor.Results H-TP could enhance the sensitivi-ty of A549/DDP cells to cisplatin by modulating the expression of apoptosis-related proteins Bax and Bcl-2,inhibiting the Keap-1/Nrf2/HO-1/GPX4 signating pathway in A549/DDP cells,and inducing ferroptosis.Conclusion H-TP enhances the sensitivity of A549/DDP cells to cisplatin by inducing the Nrf2-mediated ferroptosis pathway.

Integrins are transmembrane receptors that can coordinate signal transduction between cells and extracellular matrix or between cells.The abnormal function of integrins is one of the recognized mechanisms of tumor development.As an important regulatory mode,post-translational modification can change the conformation and physicochemical properties of proteins,thus affecting their activities,stability and functions.After the modification of the integrin,such as glycosylation and methylation,the corresponding signal transduction pathway changes,and then affects cell adhesion,migration,differentiation and other life activities,involving in diverse physiology and pathological processes.Post-translational modifications of integrins are abundant in tumor progression and play a key role in regulating the growth,metastasis and drug resistance of different tumor cells.In this review,the structure and function,post-translational modification of integrins,and their relationship with occurrence and development of tumors will be discussed,in order to provide more explorable targets for the treatment of cancer.

BACKGROUND:Ovarian tissue vitrification cryopreservation is one of the important methods for preserving fertility.Tanshinone ⅡA has various pharmacological activities,including anti-oxidation,inhibition of inflammatory response,and reduction of apoptosis,but its role as an additive for vitrification cryoprotection of ovarian tissue is still unclear.OBJECTIVE:To explore the protective effect of tanshinone ⅡA on vitrification cryopreservation of mouse ovarian tissue.METHODS:Twenty-five 6-week-old female KM mice were randomly selected and their ovarian tissues were randomly divided into five groups,with 10 ovaries per group.The fresh group was not cryopreserved.The frozen control group used vitrification cryoprotectant.The 0.5,2.5,and 5 μmol/L tanshinone ⅡA groups used vitrification cryoprotectant containing 0.5,2.5,and 5 μmol/L tanshinone ⅡA,respectively,and were cryopreserved in liquid nitrogen.After 3 days of storage,the cryopreserved tubes were taken out and thawed.The ovarian tissue and follicle morphology of each group were observed by hematoxylin-eosin staining,and the normal follicle morphology and survival rate were analyzed.The levels of superoxide dismutase,catalase,malondialdehyde,tumor necrosis factor-α,interleukin-1β,and interleukin-17 in the ovary were detected by enzyme-linked immunosorbent assay.RT-qPCR and western blot assay were used to detect the mRNA and protein expressions of nuclear factor E2-related factor 2(Nrf2)and heme oxygenase 1(HO-1)in the mouse ovary.RESULTS AND CONCLUSION:(1)Compared with the fresh group,the frozen control group had abnormal morphology of follicles at all levels in the ovary,decreased follicle survival rate(P＜0.05),decreased superoxide dismutase and catalase activities(P＜0.05);the levels of malondialdehyde,and tumor necrosis factor α,interleukin 1β,and interleukin 17 were all increased(P＜0.05),and the mRNA and protein expressions of Nrf2 and HO-1 were decreased(P＜0.05).(2)Compared with the frozen control group,different concentrations of tanshinone ⅡA could improve the morphology of follicles at all levels in the ovary,increase the survival rate of follicles,enhance the activities of superoxide dismutase and catalase,and reduce the levels of malondialdehyde,tumor necrosis factor α,interleukin 1β,and interleukin 17,increased the mRNA and protein expression of Nrf2 and HO-1 in a concentration-dependent manner,with 5 μmol/L tanshinone ⅡA having the most significant effect.(3)The results show that tanshinone ⅡA may reduce the oxidative stress level and inflammatory response of mouse ovarian tissue by mediating the Nrf2/HO-1 signaling pathway,thereby alleviating the reproductive damage caused by vitrification cryopreservation of mouse ovaries.