Objective To investigate the resistance levels of adult Culex quinquefasciatus mosquitoes to three commonly used pyrethroid insecticides in Xingyi City, Guizhou Province, as well as to examine the changes in metabolic detoxification enzyme activities and mutations in the knockdown resistance (kdr) gene thereby providing a scientific foundation for the prevention and control of Culex quinquefasciatus in the region. Methods In 2024, the larvae of Culex quinquefasciatus were collected from different locations in Xingyi and reared to adult mosquitoes in the laboratory, and their resistance to pyrethroid insecticides was determined using the adult mosquito contact tube method. Surviving samples (resistant population) from the resistance tests were examined for the activities of mixed function oxidase (MFO), glutathione-S-transferase (GST), and non-specific esterase (NSE) using a microplate reader. The genomic DNA of individual adult Culex quinquefasciatus mosquitoes was extracted, and the voltage-gated sodium channel (VGSC) gene fragment was amplified via PCR and sequenced to analyze the mutations in the kdr gene. Results The 24-h mortality rates of adult Culex quinquefasciatus exposed to 0.25% permethrin, 0.025% deltamethrin, and 0.025% beta-cypermethrin were 1.00%, 0%, and 0.88%, respectively, indicating all populations of Culex quinquefasciatus exhibited resistance to the three pyrethroid insecticides. The activity of MFO in permethrin-resistant population followed a normal distribution, while deltamethrin-resistant and cypermethrin populations exhibited skewed distributions. The resistance ratios (RRs) for the three populations were 1.17, 1.03, and 1.07 times, respectively, with no statistically significant differences in MFO activity between field populations and susceptible strains (all P>0.05). However, the GST activities of permethrin-resistant and beta-cypermethrin-resistant field populations were 1.06 and 1.45 times higher than those of sensitive strains (P<0.05). Sequencing of the VGSC gene fragments of adult Culex quinquefasciatus resistant to pyrethroid insecticides revealed that mutations occurred solely at 1014 locus, presenting two alleles: wild type TTA(L) (1.26%) and mutant TTT(F) (98.74%). Two genotypes were identified: wild/mutant heterozygotes L/F (2.52%) and mutant homozygotes F/F (97.48%). No statistically significant differences were observed in the frequency of resistance gene mutations among populations resistant to the three pyrethroid insecticides (P>0.05). Conclusion The adult mosquitoes of Culex quinquefasciatus in Xingyi have developed resistance to three commonly used pyrethroid insecticides, with changes observed in metabolic detoxification enzyme activities and mutations in the kdr gene. The resistance of Culex quinquefasciatus in Xingyi is the result of multiple mechanisms. Understanding the resistance level and mechanism can provide a reliable basis for mosquito control and resistance management. It is recommended to enhance the monitoring of mosquitoes' resistance to insecticides, prioritize environmental management focused on eradicating breeding sites, reduce the use of pyrethroid insecticides, and rotate with other classes of insecticides to delay the onset of resistance.

OBJECTIVE: To explore the neuroprotective effect and underlying mechanism of Xingnao Kaiqiao acupuncture (acupuncture for regaining consciousness and opening orifices) in the rat models of cerebral ischemia-reperfusion injury (CIRI) based on the p53 protein (p53)/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling pathway. METHODS: Of 102 male Wistar rats, 20 rats were randomly collected as a sham-operation group. Using a modified external carotid artery filament insertion method, CIRI models were prepared by occluding the middle cerebral artery in the rest rats. After modeling and excluding 1 non-successfully modeled rat and 1 dead one, the other modeled rats were randomized into a model group, an agonist group, an acupuncture group, and an acupuncture + agonist group, 20 rats in each one. Xingnao Kaiqiao acupuncture therapy was delivered in the rats of the acupuncture group and the acupuncture + agonist group. The acupoints included "Shuigou" (GV26), bilateral "Neiguan" (PC6), and "Sanyinjiao" (SP6) on the affected side. Electroacupuncture was attached to "Neiguan" (PC6) and "Sanyinjiao" (SP6) on the affected side, with dense-disperse wave, a frequency of 2 Hz/15 Hz and intensity of 1 mA. The intervention was delivered twice daily, 20 min each time and for 7 consecutive days. In the agonist group and acupuncture+agonist group, p53 agonist, COTI-2 was intraperitoneally injected (15 mg/kg), once daily for 7 consecutive days. Neurological deficit was evaluated using Zausinger's six-point scale. Cerebral infarction volume was quantified by triphenyl tetrazolium chloride (TTC) staining. Histopathological changes were observed using hematoxylin-eosin (HE) staining. Iron deposition was assessed by Prussian blue staining. Mitochondrial ultrastructure in the ischemic cortex was examined under transmission electron microscopy (TEM). Serum iron (Fe²⁺) was measured with chromometry. Malondialdehyde (MDA) and glutathione (GSH) levels in the ischemic hippocampus were determined using thiobarbituric acid and microplate assays, respectively. The mean fluorescence intensity of reactive oxygen species (ROS) in the ischemic cortex was analyzed by flow cytometry. The mRNA and protein expression of GPX4, SLC7A11, and p53 in the ischemic hippocampus were evaluated using quantitative real-time PCR (qRT-PCR) and Western blotting, respectively. RESULTS: Compared with the sham-operated group, the model group exhibited the decrease in neurological deficit score (P<0.01), and the increase in cerebral infarction volume percentage (P<0.01). The changes of brain tissue were presented in extensive cellular necrosis, pyknotic and deeply-stained nuclei, and vacuolar degeneration. The iron deposition was elevated in cortex and hippocampus (P<0.01), mitochondrial membrane density increased, the cristae was broken or reduced, and the outer membrane ruptured. The levels of Fe²⁺ and MDA, as well as the mean flourscence intensity of ROS were elevated (P<0.01) and the level of GSH was reduced (P<0.01). The mRNA and protein expression of GPX4 and SLC7A11 was reduced (P<0.01), while that of p53 rose (P<0.01). When compared with the model group, in the agonist group, the neurological deficit score was reduced (P<0.05), the percentage of infarction volume was higher (P<0.01), the histopathological damage was further exacerbated, and the percentage of iron deposition increased in the cortex and hippocampus (P<0.01). The mitochondrial quantity decreased, the membrane density increased, the mitochondrial cristae were broken or reduced, and the outer membrane was ruptured. The levels of Fe²⁺ and MDA, as well as the mean flourscence intensity of ROS were higher (P<0.01, P<0.05) and the level of GSH was reduced (P<0.05). The mRNA and protein expression of GPX4 and SLC7A11 decreased (P<0.01, P<0.05), while that of p53 was elevated (P<0.01). Besides, in comparison with the model group, the neurological deficit score was higher in the acupuncture group and the acupuncture + agonist group (P<0.01, P<0.05), the percentage of cerebral infarction volume was lower in the acupuncture group (P<0.01), the pathological damage of brain tissue was alleviated in the acupuncture group and the acupuncture + agonist group, and the percentage of iron depositiondecreased in the cortex and hippocampus (P<0.01). The mitochondrial structure was relatively clear, the mitochondrial cristae were fractured or reduced mildly in the acupuncture group and the acupuncture + agonist group. The levels of Fe²⁺ and MDA, as well as the mean flourscence intensity of ROS were lower (P<0.01) and the level of GSH was higher (P<0.01) in the acupuncture group. The mean fluorescence intensity of ROS were dropped (P<0.01) in the acupuncture + agonist group. The mRNA expression of GPX4 and SLC7A11 was elevated (P<0.01) and that of p53 was reduced (P<0.01, P<0.05) in either the acupuncture group or the acupuncture + agonist group; the protein expression of GPX4 and SLC7A11 rose (P<0.05, P<0.01) and that of p53 was dropped (P<0.01) in the acupuncture group; and the protein expression of p53 was also lower in the acupuncture + agonist group (P<0.05). When compared with the agonist group, in the acupuncture + agonist group, neurological deficit score increased (P<0.01), the percentage of cerebral infarction volume decreased (P<0.01), the pathological brain tissue damage was reduced, the percentage of iron deposition in cortex and hippocampus decreased (P<0.01), the mitochondrial structure was relatively clear and the cristae broken or reduced slightly; the levels of Fe²⁺ and MDA, as well as the mean fluorescence intensity of ROS were dropped (P<0.01), while the level of GSH increased (P<0.05); the mRNA and protein expression of GPX4 and SLC7411 was elevated (P<0.01, P<0.05), and that of p53 reduced (P<0.01). In comparison with the acupuncture + agonist group, in the acupuncture group, the neurological deficit score increased (P<0.05), the percentage of cerebral infarction volume decreased (P<0.05), the pathological brain tissue damage was alleviated, the percentage of iron deposition in cortex and hippocampus decreased (P<0.01), the mitochondrial structure was normal in tendency; the levels of Fe²⁺ and MDA, as well as the mean fluorescence intensity of ROS were reduced (P<0.05), while the level of GSH rose (P<0.01); the mRNA and protein expression of GPX4 and SLC7411 was elevated (P<0.01, P<0.05), and that of p53 reduced (P<0.01, P<0.05). CONCLUSION Xingnao Kaiqiao acupuncture can alleviate neurological damage in CIRI rats, which is obtained probably by inhibiting ferroptosis through p53/SLC7A11/GPX4 pathway.
Animals ; Reperfusion Injury/metabolism* ; Male ; Acupuncture Therapy ; Rats ; Tumor Suppressor Protein p53/genetics* ; Brain Ischemia/metabolism* ; Rats, Wistar ; Signal Transduction ; Humans ; Phospholipid Hydroperoxide Glutathione Peroxidase/genetics* ; Disease Models, Animal ; Acupuncture Points ; Amino Acid Transport System y+/genetics*

OBJECTIVE: To analyze the MYH9 gene sequence of a patient with hereditary thrombocytopenia and diffuse large B-cell lymphoma and his family members, and to explore the relationship between MYH9 gene and tumors. METHODS: Peripheral blood samples were collected from the patients and their family members for complete blood count analysis. The platelet morphology was observed under microscope. The MYH9 gene sequence was analyzed by Whole Exon Sequencing and Sanger Sequencing. RESULTS: The mutation site c.279C>A:p.(Asn93Lys) in exon 2 of the MYH9 gene were found in patient and his family members, both presenting as thrombocytopenia. The platelet count was significantly increased after the administration of Avatrombopag. CONCLUSION A novel mutation of MYH9 was found in this study, and the case was sensitive to Avatrombopag, by exploring the relationship between the MYH9 gene and tumors, suggesting that the MYH9 gene may be associated with the development of diffuse large B-cell lymphoma.
Humans ; Myosin Heavy Chains/genetics* ; Thrombocytopenia/genetics* ; Mutation ; Male ; Lymphoma, Non-Hodgkin/genetics* ; Lymphoma, Large B-Cell, Diffuse/genetics* ; Molecular Motor Proteins/genetics* ; Pedigree

Somatostatin receptor 1 (SSTR1) is a crucial therapeutic target for various neuroendocrine and oncological disorders. Current SSTR1-targeted treatments, including the first-generation somatostatin analog lanreotide (Lan) and the second-generation analog pasireotide (Pas), show promise but encounter challenges related to selectivity and efficacy. This study presents high-resolution cryo-electron microscopy structures of SSTR1 complexed with Lan or Pas, revealing the distinct mechanisms of ligand-binding and activation. These structures illustrate unique conformational changes in the SSTR1 orthosteric pocket induced by each ligand, which are critical for receptor activation and ligand selectivity. Combined with the biochemical assays and molecular dynamics simulations, our results provide a comparative analysis of binding characteristics within the SSTR family, highlighting subtle differences in SSTR1 activation by Lan and Pas. These insights pave the way for designing next-generation therapies with enhanced efficacy and reduced side effects through improved receptor subtype selectivity.

Objective: To examine the association between visceral fat metabolic levels and cardiovascular disease (CVD) among middle-aged and elderly population, so as to provide the evidence for the early identification and prevention of CVD risk in this population. Methods: Based on the database of the China Health and Retirement Longitudinal Study (CHARLS) from 2011 to 2020, baseline demographic information, lifestyle, disease history, and CVD status of participants aged ≥45 years were collected. Data on height, weight, waist circumference, and blood biochemical indicators from 2012 and 2015 were collected and used to calculate the metabolic score for visceral fat (METS-VF) and cumulative METS-VF, enabling an assessment of visceral fat metabolism levels. The K-means clustering algorithm was applied to analyze the categories of METS-VF. Multivariable logistic regression models were used to analyze the association between different METS-VF categories, cumulative METS-VF and CVD. A restricted cubic spline model was employed to examine the dose-response relationship between cumulative METS-VF and CVD. Results: A total of 3 146 participants were included, with a median age of 57.00 (interquartile range, 12.00) years. There were 1 405 males (44.66%) and 1 741 females (55.34%). METS-VF was clustered into three distinct categories: a persistently low-level group, a persistently moderate-level group, and a persistently high-level group, comprising 497, 1 302, and 1 347 individuals, accounting for 15.80%, 41.39%, and 42.82%, respectively. By the 2020 follow-up, there were 540 cases of CVD, with an overall prevalence of 17.16%. The prevalence of CVD among different METS-VF categories were 12.47%, 14.36%, and 21.60%, respectively. Multivariable logistic regression analysis showed that, after adjusting for demographic factors, lifestyle, and disease history, compared with the persistently low-level group, the persistently high-level group had a higher risk of CVD (OR=1.710, 95%CI: 1.263-2.342). Cumulative METS-VF was positively associated with CVD risk (OR=1.197, 95%CI: 1.113-1.289). Restricted cubic spline analysis indicated a linear relationship between cumulative METS-VF and CVD risk (P for nonlinearity >0.05). Conclusion Persistently high levels of METS-VF can increase the risk of CVD among middle-aged and elderly population, and there is a positive dose-response relationship between cumulative METS-VF and CVD risk.

Ubiquitination is a crucial post-translational modification process that can degrade proteins within cells and plays a vital role in maintaining protein homeostasis and abundance.Deubiquitinating enzymes(DUBs)are important proteases in the ubiquitin system.They reverse the ubiquitination process by cleaving protein chains and recycling ubiquitin molecules to regulate protein stability.Abnormal deubiquitinating enzyme activity is related to the occurrence and development of many malignant tumors.JOSD2,a DUB,is a member of the Machado-Joseph disease protein domain protease(MJD)family and characterized by a single highly conserved catalytic Josephin domain.Increasing studies have revealed a connection between JOSD2 and malignant tumors.This article elaborates on the current research status of DUBs,particularly JOSD2,in malignant tumors.Results suggest that JOSD2 is a potential target for the treatment of malignant tumors.

Standardized reporting is a crucial factor affecting the use of patient guidelines （PGs）， particularly in the reporting and presentation of recommendations. This paper introduced the current status of PG reporting， including the research on PG content and presentation formats， and provided comprehensive recommendations for PG reporting from aspects such as overall framework， recommendations， presentation format， and readability. First， the presentation of PG recommendations should include clearly defined clinical questions， recommendations and their rationale， and guidance on how patients should implement the interventions； for specific content in the PG， such as level of evidence， level of recommendation， it is recommended to explain in text the reasons for giving different levels of recommendation， i.e.， to present the logic behind giving the level of recommendation to the patient； additional information needed in the recommendation framework should be supplemented by tracing references or authoritative textbooks and literature that support the recommendations. Subsequently， the PG text should be written based on the Reporting Checklist for Public Versions of Guidelines （RIGHT-PVG） reporting framework. Finally， to enhance readability and comprehension， it is recommended to refer to the Patient Education Materials Assessment Tool （PEMAT） for translating PG content. To enhance the readability of PGs， it is suggested to present the PG content in a persona-lized and layered manner.

At present， the process and methodology of patient guidelines （PGs） development varies greatly and lacks systematic and standardised guidance. In addition to the interviews with PG developers， we have sorted out the relevant methodology for the adaptation and development of existing clinical practice guideline recommendations and facilitated expert deliberations to achieve a consensus， so as to finally put forward a proposal for guidance on the process and methodology for the development of PGs. The development of PGs can be divided into the preparation stage， the construction stage， and the completion stage in general， but the specific steps vary according to the different modes of development of PGs. The development process of Model 1 is basically the same as the patient version of the guideline development process provided by the International Guidelines Network， i.e.， team formation， screening of recommendations， guideline drafing， user testing and feedback， approval and dissemination. The developer should also first determine the need for and scope of translating the clinical practice guideline into a patient version during the preparation phase. Model 2 adds user experience and feedback to the conventional clinical practice guideline development process （forming a team， determining the scope of the PG， searching， evaluating and integrating evidence， forming recommendations， writing the guideline， and expert review）. Based on the different models， we sort out the process and methods of PG development and introduce the specific methods of PG development， including how to identify the clinical problem and how to form recommendations based on the existing clinical practice guidelines， with a view to providing reference for guideline developers and related researchers.

Objective:To establish the evaluation criteria for the quality of nursing care in orthopaedic trauma specialties, and to provide a basis for scientific evaluation of the quality of nursing care in orthopaedic trauma specialties.Methods:From February to October 2023, John Hopkinson evidence-based method was used to synthesize the evidence, clinical investigation and semi-structured interview were used to draw up the "quality standard of nursing care in orthopaedic trauma specialty", and two rounds of Delphi method of expert inquiry were used to screen, demonstrate and calibrate the evaluation standard of nursing quality in orthopaedic trauma specialty.Results:The questionnaire recovery rate of 2 rounds of expert consultation was 100.00%, the coefficient of familiarity and authority of 2 rounds of expert consultation were 0.92 and 0.93. Through 2 rounds of expert letter consultation, the quality evaluation criteria of orthopaedic trauma specialist care were established, including 3 first-level indicators (structural quality, process quality and outcome quality), 22 second-level indicators and 85 third-level indicators.Conclusions:The process of constructing the evaluation standard of nursing quality in orthopaedic trauma specialty is scientific and reliable, which can provide standard guidance for clinical practice and is conducive to the development of specialized nursing.