Objective To analyze the structural and phylogenetic characteristics of the mitochondrial genome from Bulinus globosus, so as to provide a theoretical basis for classification and identification of species within the Bulinus genus, and to provide insights into understanding of Bulinus-schistosomes interactions and the mechanisms of parasite transmission. Methods B. globosus samples were collected from the Ruya River basin in Zimbabwe. Mitochondrial DNA was extracted from B. globosus samples and the corresponding libraries were constructed for high-throughput sequencing on the Illumina NovaSeq 6000 platform. After raw sequencing data were subjected to quality control using the fastp software, genome assembly was performed using the A5-miseq and SPAdes tools, and genome annotation was conducted using the MITOS online server. Circular maps and sequence plots of the mitochondrial genome were generated using the CGView and OGDRAW software, and the protein conservation motifs and structures were analyzed using the TBtools software. Base composition and codon usage bias were analyzed and visualized using the software MEGA X and the ggplot2 package in the R software. In addition, a phylogenetic tree was created in the software MEGA X after sequence alignment with the software MAFFT 7, and visualized using the software iTOL. Results The mitochondrial genome of B. globosus was a 13 730 bp double-stranded circular molecule, containing 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and 13 protein-coding genes, with a marked AT preference. The mitochondrial genome composition of B. globosus was similar to that of other species within the Bulinus genus. Phylogenetic analysis revealed that the complete mitochondrial genome sequence of B. globosus was clustered with B. truncatus, B. nasutus, and B. ugandae into the same evolutionary clade, and gene superfamily analysis showed that the metabolism-related proteins of B. globosus were highly conserved, notably the cytochrome c oxidase family, which showed a significant consistency. Conclusions This is the first whole mitochondrial genome sequencing to decode the compositional features of the mitochondrial genome of B. globosus from Zimbabwe and its evolutionary relationship within the Bulinus genus, which provides important insights for further understanding of the phylogeny and mitochondrial genome characteristics of the Bulinus genus.

BACKGROUND:Crosslinked sodium hyaluronate gel has good mechanical property,biocompatibility,and biodegradability,and can be used as an isolated protective material in tumor radiation therapy to protect endangered organs from damage caused by excess radiation dose. OBJECTIVE:To investigate the safety and efficacy of crosslinked sodium hyaluronate gel in reducing the dose of radiation to dangerous organs during brachytherapy. METHODS:A total of 16 specific pathogen-free Kunming mice of the same age and similar body weight were selected as experimental subjects and divided into experimental group and control group by the random number table method,with 8 mice in each group.125I seeds were implanted subcutaneously in the back of mice in the experimental group,and then crosslinked sodium hyaluronate gel was injected around the radioactive particles.Only 125I seeds were implanted subcutaneously in the back of mice in the control group.After injection,the distance between the radioactive particles and the epidermis was measured by spiral CT scan,and the surface radiation dose was measured by radiation dosimeter.Within 10 weeks after injection,the growth state,survival rate,skin radiation damage,and gel retention of mice were observed. RESULTS AND CONCLUSION:(1)Spiral CT scan showed that the implanted gel was relatively concentrated and created an effective distance between the radioactive seeds and the epidermis.The body surface radiation dose of the experimental group was significantly lower than that of the control group(P<0.01).(2)During the experimental observation period,mice in both groups survived;mice in the control group showed obvious irritability and other unstable behavior in the late experimental period,and some mice in the experimental group showed similar behavior.The daily food intake of mice in the two groups had no significant change,and the body mass showed the same increasing trend.After implantation of radioactive seeds,the two groups of mice showed different degrees of radioactive skin injury.From day 23 after injection to the end of the experiment,the skin radiation injury score of the experimental group was lower than that of the control group(P<0.01).At week 10 after implantation,6 mice in the experimental group had no obvious gel residue under their skin,and 2 mice had a very small amount of scattered gel-like samples under their skin.(3)Therefore,the crosslinked sodium hyaluronate injection technique can increase the space between the radioactive target area of 125I seeds and the organ at risk outside the target through physical space occupying,which can effectively reduce the dose of the organ at risk,and play a role in the isolation and protection of the organ at risk.

Craniocervical junction zone malformations often have an insidious onset, a variety of clinical phenotypes, and are often combined with multiple malformations, making their systematic classification and staging more difficult.At present, craniocervical junction area malformations are often classified into congenital and acquired, and can also be classified into skull base malformations, atlantoaxial malformations, and cardinal malformations according to their locations. For patients with obvious occipitocervical or atlantoaxial instability, combined with symptoms of high cervical spinal cord damage, internal fixation and fusion surgery should be performed aggressively to avoid irreversible nerve damage.There is a lack of detailed categorisation and summary of the treatment of diseases associated with craniocervical junction malformations in the literature, and the treatment strategies for some of these malformations are still controversial, with different perceptions and treatment concepts in the national and international literature.

Objective To investigate the role and underlying mechanism of C1q-like protein 4 (C1ql4) in regulating the characteristics of breast cancer stem cells. Methods qRT-PCR was used to detect the expression of C1ql4 in breast cancer and normal breast epithelial cell lines, as well as to verify the transfection efficiency of C1ql4. Western blot analysis was employed to examine the phosphorylation levels of AKT, IKK, and IκB in different groups. An AKT activator was added to MDA-MB-231 cells with C1ql4 knockdown, whereas inhibitors targeting AKT, IKK, IκB, and NF-κB nuclear translocation were separately introduced to C1ql4-overexpressing MCF-7 cells. The nuclear translocation of NF-κB, expression levels of the target genes TNF-α and IL-1β, formation ability of tumorspheres, and proportion of CD44⁺/CD24^−/low stem-like subgroups were analyzed. Results C1ql4 expression in breast cancer cell lines was significantly upregulated compared with that in normal breast epithelial cells. Western blot analysis showed that p-AKT/AKT, p-IKK/IKK, and p-IκB/IκB ratios markedly reduced in C1ql4-knockdown MDA-MB-231 cells (all P<0.05) but significantly increased in C1ql4-overexpressing MCF-7 cells (all P<0.05). Rescue experiments demonstrated that the addition of an AKT activator to C1ql4-knockdown MDA-MB-231 cells resulted in the enhanced nuclear translocation of NF-κB, the increased nuclear/cytoplasmic NF-κB ratios, the elevated TNF-α and IL-1β expression levels, and significant recovery of tumorsphere formation ability and the proportion of CD44⁺/CD24^−/low stem-like subpopulations (all P<0.05). Conversely, in C1ql4-overexpressing MCF-7 cells, treatment with AKT, IKK, IκB, or NF-κB nuclear translocation inhibitors led to a reduction in NF-κB nuclear translocation, decreased nuclear/cytoplasmic NF-κB ratios, and declines in TNF-α and IL-1β expression levels, tumorsphere formation ability, and the CD44⁺/CD24^−/low subpopulation (all P<0.05). Conclusion C1ql4 promotes the translocation of NF-κB from the cytoplasm to the nucleus through the PI3K/AKT/NF-κB signaling pathway and enhances the expression of stemness in breast cancer cells.

Objective:To investigate the changes in the severity of mitral regurgitation in patients with large atrial septal defects after surgery.Methods:This study conducted a retrospective analysis using a self-controlled method on 145 patients with secundum atrial septal defects greater than 2 cm who underwent surgical treatment in the Department of Cardiology and Macrovascular Surgery at The First Affiliated Hospital of Anhui Medical University from January 2021 to October 2023. Among these patients, 74 underwent surgical closure, while 71 underwent surgical repair. Data including general information, defect size, number of surgeries of various types, preoperative transthoracic echocardiogram results, and follow-up results from the first and most recent transthoracic echocardiograms after discharge for all patients were collected. The Friedman test was used to compare the degree of mitral regurgitation before surgery and at the first and most recent follow-up after discharge. The differences in the severity of mitral regurgitation in patients before and after surgical treatment were analyzed.Results:Among the 145 patients, there were 25 males and 110 females, with an average age of (36 ± 14) years, height of (163.4 ± 8.2) cm, and body weight of (59.5 ± 10.5) kg. The diameter of the defect was (2.49 ± 0.35) cm. Before surgery, the severity of mitral regurgitation was classified as none, mild, and moderate (including mild to moderate) for 57 patients (39.3%), 83 patients (57.2%), and 5 patients (3.4%), respectively. The average follow-up duration was (324 ± 153) days. At the first follow-up after discharge, the severity of mitral regurgitation was none, mild, and moderate for 50 patients (34.5%), 87 patients (60.0%), and 8 patients (5.5%), respectively. At the most recent follow-up after discharge, the severity of mitral regurgitation was none, mild, and moderate for 30 patients (20.7%), 89 patients (61.4%), and 26 patients (17.9%), respectively. Compared with the preoperative severity of mitral regurgitation, the most recent follow-up showed worsening in 65 patients (44.8%) and improvement in 22 patients (15.2%). The mean rank sums for the three degrees of mitral regurgitation were 1.83, 1.91, and 2.26, with a statistically significant difference ( χ2 = 25.36, P = 0.001). Pairwise comparisons indicated that the most recent follow-up after discharge showed worsening compared with the preoperative severity of mitral regurgitation ( P < 0.05) and that it also worsened compared with the first follow-up after discharge ( P < 0.05). Conclusions:In patients with secundum atrial septal defects greater than 2 cm, undergoing surgery solely for atrial septal defects may result in the potential worsening of mitral regurgitation after the procedure.

Objective:To investigate the changes in the severity of mitral regurgitation in patients with large atrial septal defects after surgery.Methods:This study conducted a retrospective analysis using a self-controlled method on 145 patients with secundum atrial septal defects greater than 2 cm who underwent surgical treatment in the Department of Cardiology and Macrovascular Surgery at The First Affiliated Hospital of Anhui Medical University from January 2021 to October 2023. Among these patients, 74 underwent surgical closure, while 71 underwent surgical repair. Data including general information, defect size, number of surgeries of various types, preoperative transthoracic echocardiogram results, and follow-up results from the first and most recent transthoracic echocardiograms after discharge for all patients were collected. The Friedman test was used to compare the degree of mitral regurgitation before surgery and at the first and most recent follow-up after discharge. The differences in the severity of mitral regurgitation in patients before and after surgical treatment were analyzed.Results:Among the 145 patients, there were 25 males and 110 females, with an average age of (36 ± 14) years, height of (163.4 ± 8.2) cm, and body weight of (59.5 ± 10.5) kg. The diameter of the defect was (2.49 ± 0.35) cm. Before surgery, the severity of mitral regurgitation was classified as none, mild, and moderate (including mild to moderate) for 57 patients (39.3%), 83 patients (57.2%), and 5 patients (3.4%), respectively. The average follow-up duration was (324 ± 153) days. At the first follow-up after discharge, the severity of mitral regurgitation was none, mild, and moderate for 50 patients (34.5%), 87 patients (60.0%), and 8 patients (5.5%), respectively. At the most recent follow-up after discharge, the severity of mitral regurgitation was none, mild, and moderate for 30 patients (20.7%), 89 patients (61.4%), and 26 patients (17.9%), respectively. Compared with the preoperative severity of mitral regurgitation, the most recent follow-up showed worsening in 65 patients (44.8%) and improvement in 22 patients (15.2%). The mean rank sums for the three degrees of mitral regurgitation were 1.83, 1.91, and 2.26, with a statistically significant difference ( χ2 = 25.36, P = 0.001). Pairwise comparisons indicated that the most recent follow-up after discharge showed worsening compared with the preoperative severity of mitral regurgitation ( P < 0.05) and that it also worsened compared with the first follow-up after discharge ( P < 0.05). Conclusions:In patients with secundum atrial septal defects greater than 2 cm, undergoing surgery solely for atrial septal defects may result in the potential worsening of mitral regurgitation after the procedure.

To identify clinical viral diseases characterized by reproductive disorders and abortion,a quadruple qPCR method was established for simultaneous detection of PRV,PPV,PCV2 and AS-FV.Four pairs of specific primers and probes were designed according to the conserved genes of four viruses in the NCBI gene bank.The annealing temperature,primer concentration and probe concentration of the reaction were optimized,and the specificity,sensitivity and repeatability of the method were tested.The results showed that the method could not detect other pathogens except the target ones.The minimum detection limit of PRV,PPV,PCV2 and ASFV was 10 copies.Intra-group and inter-group repeatability tests showed that the coefficient of variation of C,values be-tween different batches was less than 3％,indicating that the method was highly specific,sensitive and stable.Establishment of an efficient and sensitive quadruple qPCR method provides technical reference for the clinical prevention and control of porcine pseudorabies virus disease,porcine circo-virus disease,porcine parvovirus disease and African swine fever.

Craniocervical junction zone malformations often have an insidious onset, a variety of clinical phenotypes, and are often combined with multiple malformations, making their systematic classification and staging more difficult.At present, craniocervical junction area malformations are often classified into congenital and acquired, and can also be classified into skull base malformations, atlantoaxial malformations, and cardinal malformations according to their locations. For patients with obvious occipitocervical or atlantoaxial instability, combined with symptoms of high cervical spinal cord damage, internal fixation and fusion surgery should be performed aggressively to avoid irreversible nerve damage.There is a lack of detailed categorisation and summary of the treatment of diseases associated with craniocervical junction malformations in the literature, and the treatment strategies for some of these malformations are still controversial, with different perceptions and treatment concepts in the national and international literature.

To establish a TaqMan-based multiplex RT-qPCR method for the identification of Japa-nese encephalitis virus(JEV),Porcine reproductive and respiratory syndrome virus(PRRSV),and Classical swine fever virus(CSFV),this study designed and synthesized three pairs of specific primers and probes based on the conserved sequences of JEV E,PRRSV ORF6,and CSFV E2 a-vailable in the NCBI GenBank.By optimizing the reaction system and protocol,a multiplex RT-qPCR method for detecting these three viruses was developed and applied to the detection of clini-cal samples.The results showed that the established TaqMan multiplex RT-qPCR specifically am-plified the gene fragments of JEV,PRRSV,and CSFV,and did not amplify other non-target genes,indicating good specificity of the method.Intra-assay and inter-assay repeatability tests showed that the coefficient of variation(Cv)values were all below 3％,demonstrating that the method has ex-cellent repeatability.Sensitivity tests revealed that the minimum detectable amount for the recom-binant plasmids of the three viruses was 100 copies/pL.Using the established method,a total of 969 samples,including blood,aborted fetuses,semen,and deceased pigs,from 26 pig farms in Guizhou Province were tested.The detection rates were 34.3％(332/969)for JEV,28.3％(274/969)for PRRSV,and 19.8％(192/969)for CSFV.The co-infection rates were 10.1％(98/969)for JEV and PRRSV,12.1％(117/969)for JEV and CSFV,and 14.6％(141/969)for CSFV and PRRSV.Additionally,the triple co-infection rate of JEV,PRRSV,and CSFV was 7.9％(77/969).These results indicate that the TaqMan multiplex RT-qPCR method developed in this study is ef-fective for detecting these three viruses in pig farms,providing technical support for identifying vi-ral causes of reproductive disorders.

To identify clinical viral diseases characterized by reproductive disorders and abortion,a quadruple qPCR method was established for simultaneous detection of PRV,PPV,PCV2 and AS-FV.Four pairs of specific primers and probes were designed according to the conserved genes of four viruses in the NCBI gene bank.The annealing temperature,primer concentration and probe concentration of the reaction were optimized,and the specificity,sensitivity and repeatability of the method were tested.The results showed that the method could not detect other pathogens except the target ones.The minimum detection limit of PRV,PPV,PCV2 and ASFV was 10 copies.Intra-group and inter-group repeatability tests showed that the coefficient of variation of C,values be-tween different batches was less than 3％,indicating that the method was highly specific,sensitive and stable.Establishment of an efficient and sensitive quadruple qPCR method provides technical reference for the clinical prevention and control of porcine pseudorabies virus disease,porcine circo-virus disease,porcine parvovirus disease and African swine fever.