ObjectiveTo investigate the influence of histone deacetylase 3 (HDAC3) on the occurrence, development of psoriasis-like inflammation in mice, and the relative immune mechanisms. MethodsHealthy C57BL/6 mice aged 6-8 weeks were selected and randomly divided into 3 groups: control group (Control), psoriasis model group (IMQ), and HDAC3 inhibitor RGFP966-treated psoriasis model group (IMQ+RGFP966). One day prior to the experiment, the back hair of the mice was shaved. After a one-day stabilization period, the mice in Control group was treated with an equal amount of vaseline, while the mice in IMQ group was treated with imiquimod (62.5 mg/d) applied topically on the back to establish a psoriasis-like inflammation model. The mice in IMQ+RGFP966 group received intervention with a high dose of the HDAC3-selective inhibitor RGFP966 (30 mg/kg) based on the psoriasis-like model. All groups were treated continuously for 5 d, during which psoriasis-like inflammation symptoms (scaling, erythema, skin thickness), body weight, and mental status were observed and recorded, with photographs taken for documentation. After euthanasia, hematoxylin-eosin (HE) staining was used to assess the effect of RGFP966 on the skin tissue structure of the mice, and skin thickness was measured. The mRNA and protein expression levels of HDAC3 in skin tissues were detected using reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB), respectively. Flow cytometry was employed to analyze neutrophils in peripheral blood and lymph nodes, CD4⁺ T lymphocytes, CD8⁺ T lymphocytes in peripheral blood, and IL-17A secretion by peripheral blood CD4⁺ T lymphocytes. Additionally, spleen CD4⁺ T lymphocyte expression of HDAC3, CCR6, CCR8, and IL-17A secretion levels were analyzed. Immunohistochemistry was used to detect the localization and expression levels of HDAC3, IL-17A, and IL-10 in skin tissues. ResultsCompared with the Control group, the IMQ group exhibited significant psoriasis-like inflammation, characterized by erythema, scaling, and skin wrinkling. Compared with the IMQ group, RGFP966 exacerbated psoriasis-like inflammatory symptoms, leading to increased hyperkeratosis. The psoriasis area and severity index (PASI) skin symptom scores were higher in the IMQ group than those in the Control group, and the scores were further elevated in the IMQ+RGFP966 group compared to the IMQ group. Skin thickness measurements showed a trend of IMQ+RGFP966>IMQ>Control. The numbers of neutrophils in the blood and lymph nodes increased sequentially in the Control, IMQ, and IMQ+RGFP966 groups, with a similar trend observed for CD4⁺ and CD8⁺ T lymphocytes in the blood. In skin tissues, compared with the Control group, the mRNA and protein levels of HDAC3 decreased in the IMQ group, but RGFP966 did not further reduce these expressions. HDAC3 was primarily located in the nucleus. Compared with the Control group, the nuclear HDAC3 content decreased in the skin tissues of the IMQ group, and RGFP966 further reduced nuclear HDAC3. Compared with the Control and IMQ groups, RGFP966 treatment decreased HDAC3 expression in splenic CD4⁺ and CD8⁺ T cells. RGFP966 treatment increased the expression of CCR6 and CCR8 in splenic CD4⁺ T cells and enhanced IL-17A secretion by peripheral blood and splenic CD4⁺ T lymphocytes. Additionally, compared with the IMQ group, RGFP966 reduced IL-10 protein levels and upregulated IL-17A expression in skin tissues. ConclusionRGFP966 exacerbates psoriatic-like inflammatory responses by inhibiting HDAC3, increasing the secretion of the cytokine IL-17A, and upregulating the expression of chemokines CCR8 and CCR6.

Heart and spleen deficiency syndrome is the most common syndrome type in patients with insomnia. Based on the theory of disease syndrome-combined animal model， this paper used multiple databases to search for the keywords "heart and spleen deficiency"， "insomnia"， "sleepless"， "disease syndrome-combined animal model"， "model evaluation"， etc. It selected the literature related to the animal model of insomnia with heart and spleen deficiency in the past 20 years to evaluate from the aspects of model establishment， modeling factors， syndrome model， disease model， macro characterization & macro characterization evaluation scale， micro indicators， etc. It is found that the existing animal model of insomnia with heart and spleen deficiency is not completely constructed by the method of disease syndrome combination of disease modeling factors and syndrome modeling factors. In the model using this method， the single establishment factor of heart and spleen deficiency does not conform to the clinical reality of disease， and the selection of the factors for the insomnia model is not closely related to or even separated from the syndrome performance. There is a problem of insufficient quantification of macro representation when the macro representation of the model replaces the symptoms related to heart and spleen deficiency syndrome and insomnia in an equivalent manner for macro representation evaluation， which can be improved according to the quantitative ideas and examples of the existing macro representation and macro representation evaluation scale. There are few specific indicators of heart and spleen deficiency syndrome in micro indicators. The micro research of heart and spleen deficiency syndrome and the essence of other traditional Chinese medicine （TCM） syndromes can be carried out by metabonomics and other technologies combined with the theory of corresponding prescription and syndrome， along the specific related ideas of "prescription and syndrome， treatment principle and selection of prescription， treatment principle and selection of acupoints， as well as therapeutic mechanism and syndrome essence". The future users and researchers of animal models of insomnia with heart and spleen deficiency can get improved methods and ideas through the shortcomings of animal models of heart and spleen deficiency listed in this paper and construct animal models of insomnia with heart and spleen deficiency that are more suitable for clinical practice， so as to establish a more perfect modeling method and evaluation system of disease syndrome-combined animal model.

Heart and spleen deficiency syndrome is the most common syndrome type in patients with insomnia. Based on the theory of disease syndrome-combined animal model， this paper used multiple databases to search for the keywords "heart and spleen deficiency"， "insomnia"， "sleepless"， "disease syndrome-combined animal model"， "model evaluation"， etc. It selected the literature related to the animal model of insomnia with heart and spleen deficiency in the past 20 years to evaluate from the aspects of model establishment， modeling factors， syndrome model， disease model， macro characterization & macro characterization evaluation scale， micro indicators， etc. It is found that the existing animal model of insomnia with heart and spleen deficiency is not completely constructed by the method of disease syndrome combination of disease modeling factors and syndrome modeling factors. In the model using this method， the single establishment factor of heart and spleen deficiency does not conform to the clinical reality of disease， and the selection of the factors for the insomnia model is not closely related to or even separated from the syndrome performance. There is a problem of insufficient quantification of macro representation when the macro representation of the model replaces the symptoms related to heart and spleen deficiency syndrome and insomnia in an equivalent manner for macro representation evaluation， which can be improved according to the quantitative ideas and examples of the existing macro representation and macro representation evaluation scale. There are few specific indicators of heart and spleen deficiency syndrome in micro indicators. The micro research of heart and spleen deficiency syndrome and the essence of other traditional Chinese medicine （TCM） syndromes can be carried out by metabonomics and other technologies combined with the theory of corresponding prescription and syndrome， along the specific related ideas of "prescription and syndrome， treatment principle and selection of prescription， treatment principle and selection of acupoints， as well as therapeutic mechanism and syndrome essence". The future users and researchers of animal models of insomnia with heart and spleen deficiency can get improved methods and ideas through the shortcomings of animal models of heart and spleen deficiency listed in this paper and construct animal models of insomnia with heart and spleen deficiency that are more suitable for clinical practice， so as to establish a more perfect modeling method and evaluation system of disease syndrome-combined animal model.

Objective To analyze the clinical characteristics of recipients with interstitial lung disease (ILD) who underwent second lung transplantation and summarize the diagnostic and therapeutic experience. Methods A retrospective analysis was conducted on the clinical data of 14 patients who underwent first and second lung transplants for ILD at the First Affiliated Hospital of Guangzhou Medical University from January 2015 to December 2024. The preoperative conditions, intraoperative events, postoperative treatments and prognoses of the first and second lung transplantation were compared, and the postoperative survival of ILD patients after the second lung transplantation was analyzed. Results Among the 14 recipients of the second lung transplant, 13 underwent the procedure due to chronic lung allograft dysfunction, and 1 due to airway complications. The median interval time from the first to the second lung transplant was 32 (2, 80) months. Before the second transplantation, 2 recipients required endotracheal intubation and mechanical ventilation, and 2 required endotracheal intubation, mechanical ventilation, and extracorporeal membrane oxygenation (ECMO) support. The surgical time for the second lung transplantation was longer than that for the first, with increased intraoperative red blood cell and plasma transfusion volumes, the proportion of ECMO support during the second lung transplantation was higher than that during the first (all P<0.05). However, the cold ischemia time for one-sided lung transplant completion in the first lung transplant was similar to that in the second lung transplantation (P>0.05). The median follow-up time after the second lung transplantation was 32 (1, 63) months. The 1-month, 6-month and 1-year survival rates after the second lung transplantation were 79%, 57% and 50%, respectively, with causes of death being infection, multiple organ failure and gastrointestinal bleeding. Conclusions For ILD patients undergoing second lung transplantation after the first lung transplantation, the second lung transplantation is more challenging, with longer surgical time and higher intraoperative blood loss. It requires higher surgical skills and perioperative management. Non-emergency second transplantation may still achieve good results.

Objective To compare the clinical efficacies of unilateral biportal endoscopy(UBE)technique and percutaneous endoscopic lumbar discectomy(PELD)technique in the treatment of lumbar disc herniation.Methods The clinical data of 149 patients with lumbar disc herniation in our hospital were retrospectively analyzed and divided into the UBE group(n=80)and the PELD group(n=69)according to different surgical methods.The operation time,intraoperative fluoroscopy frequency,intraoperative blood loss,hospital stay,postoperative complications,visual analogue scale(VAS)score,Oswestry disability index(ODI)score,intervertebral disc height and vertebral canal area of the two groups were compared.Results The operation time in the UBE group was longer than that in the PELD group,and the intraoperative fluoroscopy frequency was fewer than that in the PELD group,the differences were statistically significant(P<0.001).There was no significant difference in the intraoperative blood loss or hospital stay between the two groups(P>0.05).There was no significant difference in the VAS or ODI scores at each time point between the two groups(P>0.05).There was no significant difference in the intervertebral disc height or vertebral canal area at each time point between the two groups(P>0.05).The postoperative vertebral canal areas of patients in the two groups were greater than those before surgery,and the differences were statistically significant(P<0.05).The incidence of postoperative complications in the UBE group was lower than that in the PELD group,the difference was statistically significant(P<0.05).Conclusion In terms of short-term efficacy,both PELD technique and UBE technique can effectively relieve the symptoms of low back and leg pain caused by lumbar disc herniation,and the UBE technique has longer operation time,but with fewer intraoperative fluoroscopy frequency,and lower incidence of postoperative complications.

Objective:KPC-2-producing Klebsiella pneumoniae is able to cause drug resistance to β-lactam antibiotics and car-bapenems antibiotics.Phage display technology was used to screen nanobodies that specifically bind to KPC-2 from the anti-KPC-2 nanobodies library,which provided technical supports for the detection and diagnosis of drug resistance of KPC-2-producing Klebsiella pneumoniae.Methods:A bimodal camel was firstly immunized by the recombinant KPC-2,then,RNA was extracted from peripheral blood lymphocytes of the bimodal camel and reverse transcribed into cDNA.Nanobody fragments were amplified by two rounds of nested PCR,and the antibody library were constructed.The specific nanobodies were screened by the phage display technology.Lastly the epitope analysis and affinity determination were performed by HPLC and OCTET,respectively.Results:A nanobody library was con-structed with a capacity of 5.47×108 cfu/ml and the inserting effective fragments of not less than 81.25%;the immune elutriation method of anti-KPC-2 nanobody was established.Two nanobodies,K2 and K5,with different CDR3 regions were obtained,and their affinities were 6.0 nmol/L and 4.8 nmol/L,respectively.Moreover,K2 and K5 were non-competitive for binding epitopes on KPC-2.Conclu-sion:Two specific nanobodies with different epitopes were successfully panned out,which expect to replace traditional antibodies for detection and diagnosis of Klebsiella pneumoniae drug-resistant diseases.

This study established an efficient and specific method for type analysis of genetic variants of Balantioides coli.The restriction endonucleases ApoI and PflMI were selected to digest the PCR amplification products of ITS1-5.8S rDNA-ITS2 of B.coli,and to establish a PCR-RFLP typing method.The PCR-RFLP method was subsequently used to analyze the genetic variants in clinical fecal samples from pigs,sheep,and guinea pigs.The PCR-RFLP method based on ApoI and PflMI accurately distinguished the main A and B genetic variants of B.coli,and further divided the main B type into B-c and B-t sub-types of genetic variants with PflMI.Compared with the results of microscopy and sequencing,the PCR-RFLP method showed good specificity and higher sensitivity,and was able to identify not only single but also multiple variants of B.coli in a single clinical sample.This study successfully established the PCR-RFLP method for B.coli,which can be used for genetic di-versity identification and molecular epidemiological studies of B.coli.

The advancement of Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)gene editing technology has revolutionized the comprehension of human genome,propelling molecular and cellular biology research into unexplored realms and accelerating progress in life sciences and medicine.CRISPR-based gene screening,recognized for its efficiency and practicality,is widely utilized across diverse biological fields.Aging is a multifaceted process governed by a myriad of genetic and epigenetic factors.Unraveling the genes regulating aging holds promise for understanding this intricate phenomenon and devising strategies for its assessment and intervention.This review provides a comprehensive overview of the progress in CRISPR screening and its applications in aging research,while also offering insights into future directions.CRISPR-based genetic-manipulation tools are positioned as indispensable instruments for mitigating aging and managing age-related diseases.

Objective:To study the serological characteristics of ABO*A2.08 subtype and explore its genetic molecular mechanism.Methods:ABO blood group identification was performed on proband and her family members by routine serological methods.ABO genotyping and sequence analysis were performed by polymerase chain reaction-sequence specific primer(PCR-SSP),and direct sequencing of PCR products from exons 6 and 7 of ABO gene were directly sequenced and analyzed.The effect of gene mutation in A2.08 subtype on structural stability of GTA protein was investigated by homologous protein conserved analysis,3D molecular modeling and protein stability prediction.Results:The proband's serological test results showed subtype Ax,and ABO genotyping confirmed that the proband's genotype was ABO*A207/08.Gene sequencing of the proband's father confirmed the characteristic variation of c.539G＞C in the 7th exon of ABO gene,leading to the replacement of polypeptide chain p.Arg180Pro(R180P).3D protein molecular modeling and analysis suggested that the number of hydrogen bonds of local amino acids in the protein structure was changed after the mutation,and protein stability prediction showed that the mutation had a great influence on the protein structure stability.Conclusion:The mutation of the 7th exon c.539G＞C of ABO gene leads to the substitution of polypeptide chain amino acid,which affects the structural stability of GTA protein and leads to the change of enzyme activity,resulting in the A2.08 phenotype.The mutated gene can be stably inherited.

Purpose: This study evaluated the value of contrast-enhanced ultrasonography (CEUS) in the ultrasound-guided microwave ablation (MWA) treatment of symptomatic focal uterine adenomyosis. Methods: This retrospective study was conducted between March 2020 and January 2023, enrolling 52 patients with symptomatic focal uterine adenomyosis who had undergone MWA. All patients were examined with CEUS before and after MWA. The non-perfused volume (NPV) was compared between CEUS and dynamic contrast-enhanced magnetic resonance imaging (DCEMRI) following ablation. Therapeutic efficacy and safety were evaluated at 3-, 6-, and 12-month follow-ups. Additionally, this study explored the correlations between pre-treatment CEUS features and a volume reduction ratio indicating sufficient ablation, defined as 50% or more at the 3-month follow-up. Results: No significant differences in NPV were noted between CEUS and DCE-MRI immediately after MWA and during follow-up (all P>0.05). At the 3-month follow-up, the median VRRs for the uterus and adenomyosis were 33.2% and 63.9%, respectively. Sufficient ablation was achieved in 69.2% (36/52) of adenomyosis cases, while partial ablation was observed in the remaining 30.8% (16/52). The identification of non-enhancing areas on pre-treatment CEUS was associated with sufficient ablation (P=0.016). At the 12-month follow-up, significant decreases were observed in both the uterine and adenomyosis volumes (all P<0.001). Dysmenorrhea and menorrhagia were significantly alleviated at 12 months, and no major complications were encountered. Conclusion CEUS can be used to evaluate the ablation zone of focal adenomyosis that has been treated with MWA, similarly to DCE-MRI. The identification of non-enhancing areas on pretreatment CEUS indicates satisfactory treatment outcomes.