To summarize the distribution characteristics of human papillomavirus(HPV) infection types in patients with cervical squamous intraepithelial lesion(SIL) and cervical cancer(CC), and to explore the impact of HPV vaccination, HPV infection types, and general clinical data on different grades of cervical lesions.

Background Amid global climate change, extreme environmental events are occurring more frequently, and it is imperative to investigate the impacts of combined exposure to fine particluate matter (PM_2.5) and cold spells (CS) on population mortality. Objective To analyze the association between sequential extreme PM_2.5-cold spell (EP-CS) events and non-accidental mortality among residents in Zigong City from 2016 to 2021. Methods Using time-series study design, meteorological data in Zigong were collected from the Zigong Meteorological Bureau for the period from January 1, 2016 to December 31, 2021, while daily non-accidental mortality data were obtained from the mortality surveillance system of the Zigong Center for Disease Control and Prevention. We adopted the percentile method to define extreme PM_2.5 events and cold spells. We analyzed the risk effect of EP-CS events on non-accidental mortality among residents in this city and explored the potential amplification of damage resulting from different patterns of consecutive extreme events by using distributed lag nonlinear model (DLNM). We also conducted stratified analyses based on age, gender, education level, and marital status. Results The EP-CS events demonstrated a significant impact on non-accidental mortality among the local residents, exhibiting a certain lagged effect. The effects on the overall residents lasted from lag0 (RR=1.030, 95%CI: 1.013, 1.048) to lag14 (RR=1.035, 95%CI: 1.019, 1.052). Notably, the effects were more pronounced among females, individuals aged 65 years and above, and those who were never married, divorced, or widowed. Different patterns of EP-CS events all associated with adverse effects, the health impact of EP-CS events was significantly greater than that of individual PM_2.5 pollution or CS events. The analysis of lag effects across different event patterns revealed that the overall effect of EP-CS events with shorter intervals (0–7 d) had a stronger effect compared to EP-CS with longer intervals (8–14 d), and the RR values of lag14 were 1.034 (95%CI: 1.015, 1.054) and 1.017 (95%CI: 1.007, 1.027), suggesting that the damaging effect of compound events occurring in the short term was more significant. Conclusion All sequential extreme EP-CS events have an impact on non-accidental mortality among residents in this city, with compound events demonstrating a stronger effect. Females, individuals aged ≥65 years, and those who were never married, divorced, or widowed are more sensitive to EP-CS events.

This study aims to establish an indirect ELISA method for detecting RVFV antibodies u-sing recombinant proteins of Rift Valley fever virus(RVFV)Gn protein Ⅱ-Ⅲ structural domains as the encapsulated antigen which was expressed by the Escherichia coli(E.coli)expression sys-tem.The gene sequences encoding the Ⅱ and Ⅲ subdomains of RVFV Gn protein were inserted in-to pET-30a(+)to construct the recombinant plasmid pET-RVFV Gn-D Ⅱ-Ⅲ.After transforma-tion of the recombinant plasmid into DE3(BL21)competent cells,the recombinant Gn-D Ⅱ-Ⅲ protein was induced with IPTG and purified using affinity chromatography.An indirect ELISA method for the detection of RVFV antibodies was developed using purified recombinant protein as coating antigen and SPA-HRP as the enzyme-labelled secondary antibody.Western blot analysis confirmed that the RVFV Gn-D Ⅱ-Ⅲ protein was successfully expressed.The optimal expression conditions for RVFV Gn-D Ⅱ-Ⅲ protein were induced with 0.8 mmol/L IPTG at 37 ℃ for 5 h.The Gn-D Ⅱ-Ⅲ protein was purified using affinity chromatography with a purity of 91.9％,and the purified protein was used as the encapsulated antigen to develop an ELISA assay for RVFV anti-bodies.The specificity evaluation showed that the method specifically detected RVFV-positive sera and did not cross-react with sera positive for West Nile virus(WNV),Ebola virus(EBOV),Mar-burg virus(MARV)and tick-borne encephalitis virus(TBEV).When the RVFV Gn-D Ⅲ-Ⅲ posi-tive serum was diluted to 6 400 times,the test result still showed positive results,demonstrating the method had good sensitivity.The repeatability evaluation results indicated that the variation co-efficients for both intra-and inter-batch responses was less than 10％,indicating that the method had good repeatability.In conclusion,the RVFV Gn-D Ⅱ-Ⅲ protein was successfully expressed u-sing the E.coli expression system.The purified recombinant Gn-D Ⅱ-Ⅲ protein was used as the encapsulated antigen to develop an indirect ELISA assay for RVFV antibodies,which provides a preliminary basis for the diagnosis of RVF and the research and development of RVF vaccines.

To establish an indirect enzyme linked immunosorbent assay(ELISA)method for the detection of Zaire Ebola virus(ZEBOV)specific antibodies,the full-length of ZEBOV GP1 gene was amplified by PCR and cloned into pET-30a(+)vector to generate the pET-30a(+)-GP1 plasmid.After expressed in the E.coli expression system,the purified GP1 protein was used as coating antigen to establish the indirect ELISA method for detection of ZEBOV antibody.The con-ditions including concentration of coating antigen and serum dilution were determined by chess-board titration.Specificity,sensitivity,and reproducibility of the established ELISA detection meth-od were evaluated.GP1 protein was successfully prepared by prokaryotic expression,and was used as the coatingantigen for indirect ELISA.By optimizing the reaction conditions,the optimal concen-tration of the coating antigen was determined to be 0.5 g/L;the optimal dilution of serum was cal-culated to be 1∶3 200;the optimal dilution of enzyme-labeled secondary antibody was measured to be 1∶20 000.The established method exhibited excellent specificity,sensitivity,and reproducibili-ty.In the present study,the GP1 protein was successfully expressed in the E.coli expression sys-tem and the high purity GP1 protein was used as the coating protein to establish an indirect ELISA assay for ZEBOV antibody.This method is highly specific,sensitive,and reproducible,which provides technical support for the fur-ther study of the biological function of GP1 and the detection of ZEBOV antibody in serum.

Feline herpesvirus type Ⅰ(FHV-1)was used as the vector.The gI and gE genes of FHV-1 were replaced with the feline calicivirus(FCV)VP1 gene and the red fluorescent protein(mCherry)gene by CRISPR/Cas9 systems and homologous recombination technology,and the re-combinant virus strain FHV △gI&gE/VP1-mCherry+was successfully rescued.The recombinant virus strain was purified by plaque assay.The biological characteristics and genetic stability of the recombinant virus were analyzed by indirect immunofluorescence assay,plaque morphological anal-ysis,and PCR.The results of the indirect immunofluorescence identification showed that the re-combinant virus FHV △gI&gE/VP1-mCherry+could express the VP1 protein in F81 cells,and the growth characteristics of the recombinant virus were not significantly different from those of the parent virus FHV-1.The plaque morphology and staining results indicated that the area of the plaque formed by the recombinant virus was smaller than that of the parent virus,suggesting that the spread ability of the recombinant virus between cells was reduced after the deletion of the gI and gE genes.The result of PCR showed that the VP1 gene could still be detected after 15 succes-sive passages of the recombinant virus,indicating that the recombinant virus had good genetic stability.In this study,the recombinant virus strain expressing the FCV VP1 protein was successfully prepared,which will lay a foundation for the development of engineered FCV and FHV-1 vaccine.

Objective:To explore the heterogeneity in the trajectories ofperinatal depressive symptoms and identify the related factors.Methods:A longitudinal follow-up study was conducted from January 2020 to June 2021.Baseline information,marriage satisfaction and social support status were collected at enrolment.Perinatal de-pressive symptoms were measured using the Edinburgh Postnatal Depression Scale(EPDS)at five time-points from second trimester to three-months postpartum.A Latent Class Growth Model(LCGM)was used to identify sub-groups of perinatal depressive symptoms trajectories and logistic regression was applied to examine associated risk and protective factors.Results:Two latent depression trajectory subgroups were identified A majority(72.2％,n=1 023)followed a curvilinear trajectory with low baseline scores that increased over time(curvilinear group),while 27.8％(n=393)maintained high,stable scores(linear group).Older age,higher marital satisfaction,and stronger social support were associated with a lower likelihood of belonging to the linear group(OR=0.96,0.94,0.89,re-spectively),while a personal or family history of depression increased this likelihood(OR=2.50,6.51).Conclusion:Trajectories of perinatal depressive symptoms vary significantly among individuals.The risk of persis-tent high-level symptoms is lowered by older age,stronger marital satisfaction,and greater social support,and heightened by a personal or family history of depression.

Objective:To explore the method for selecting characteristic prices of Chinese patent medicines based on informa-tion entropy theory.It involves analyzing the connotative differences among various price indicators and utilizing information entropy metrics to validate the scientific rigor of characteristic price selection so as to optimize the pricing model for Chinese patent medi-cines and improve the accuracy of price evaluation.Methods:A correlation analysis and information entropy calculation are con-ducted on the median price of the smallest preparation unit,average daily cost,and average course cost of TCM.It compares the information diversity and uncertainty of different pricing indicators.Results:The average daily cost exhibits the highest information diversity and uncertainty among all the pricing indicators examined.Conclusion:It is recommended that the average daily cost be used as the dependent variable for characteristic prices in TCM pricing research.This choice plays an important role in optimizing TCM pricing models and enhancing the accuracy of price evaluation.

Objective:To establish a scientific,systematic,and objective indicator system for the characteristic price variables of Traditional Chinese Patent Medicines(TCPM),providing a reference framework for the pricing mechanism of TCPM.Methods:The brainstorming method was initially used to screen related variable indicators.The Nominal Group Technique(NGT)and Delphi methods were applied to gather expert opinions,and SPSS 28.0 was employed for data statistical analysis.It led to the development of a TCPM characteristic price variable indicator system consisting of 6 dimensions,14 characteristic variables and 26 measurement indicators.Results:The authority coefficient of the experts exceeded 0.7,indicating the representativeness of the results.Expert opinions were generally concentrated.Based on the collected opinions and statistical analysis,the scope of selected TCPM characteristic price variables was preliminarily established.Conclusion:The TCPM characteristic price variable indicator system was initially developed.However,due to the complexity of the pricing mechanism and divergent expert opinions,further qualitative and quantitative research methods,along with a dynamic adjustment mechanism,are needed to verify and refine the system.

The National Essential Medicines System could protect public health and ensure access to essential medications.Although the current selection methods for China's National Essential Medicines Lists(NEMLs)are becoming more scientific and standardized,there are still problems such as much emphasis on expert experience and the lack of transparency of decision-making basis.To address these issues,it proposes an evidence-based decision-making pathway for NEMLs selection guided by clinical value.This approach ensures a strong integration of evidence and decision-making,offering valuable insights for improving the adjustment procedures and selection criteria of the NEMLs in China.

Objective To investigate the effect and mechanism of nodakenin(Nod)in neuropathic pain(NP).Methods Differential expression genes in the primary somatsensory cortex(S1)of NP data and overlapping genes between the dataset and mitochondrial data were screened and analyzed.Overlapping gene interaction networks were overlapped and core genes were screened.A total of 27 mice were randomly divided into the sham operation group,the model group and the drug administration group(9 mice/group).The chronic compression injury model of sciatic nerve was constructed in the model group and the drug administration group.Nod 10 mg/kg was intraperitoneally injected into the drug administration group for 1 week.Changes of pain behavior and motor ability in mice were detected.HE staining and Nissl staining were used to detect effects of nerve injury and inflammation on brain tissue of S1 region of mice.The expression levels of interleukin-1β,early gene(c-Fos),panthenol-cytochrome c reductase complex III subunit(Uqcrq)and ubiquinone oxidoreductase subunit(Nduf)b5 in S1 brain region were analyzed by Western blot assay.Molecular docking was used to study the target of Nod.PC12 cells were divided into the control group,the IL-1β group(1 μmol/L IL-1β treatment)and the IL-1β+Nod group(1 μmol/L IL-1β+1 μmol/L Nod treatment),and mitochondrial membrane potential was detected in each group.Results In the NP dataset GSE180627,S1 brain region contained 293 differentially expressed genes,and the mitochondrial data contained 1 082 genes.There were 34 overlapping genes,and genes related to oxidative phosphorylation and electron transport chain were enriched.The protein interaction network showed that core genes included electron transport chain related proteins Ndufb5,Uqcrq,Ndufs8,Ndufa7,Ndufa3,Cox6b1 and Mrps33.Compared with the model group,the mechanical foot shrinkage threshold,thermal foot shrinkage reflex latency and rod rotation residence time of mice were increased in the drug administration group,the number of inflammatory infiltrating cells in S1 tissue and the number of Nislet bodies in neurons,expression levels of c-Fos and IL-1β in neurons were decreased,and expression levels of Uqcrq and Ndufb5 were increased(P＜0.05).Molecular docking showed that Nod could bind Uqcrq and Ndufb5.Compared with the IL-1β group,the fluorescence signal of mitochondrial membrane potential was enhanced in the IL-1β+Nod group(P＜0.05).Conclusion Nodakenin can improve pain behavior in mice,and its mechanism involves ameliorating mitochondrial damage in S1.