Objective : To construct a knockdown recombinant adeno⁃associated virus (AAV⁃shCTSK) targeting the mouse Cathepsin K(CTSK) gene and assess the knockdown efficiency of AAV⁃shCTSK in mice , and to investigate its impact on lipid storage within adipose tissue. Methods : ShRNA primers specific to both the negative control (NC) and CTSK were designed , annealed , and integrated into the backbone vector. Following clone selection and sequencing for validation , recombinant plasmids were purified. Adeno⁃associated viral vectors , along with packaging and helper plasmids , were co⁃transfected into 293T cells using the transfection reagent PEI for adeno⁃associat⁃ed viral packaging and amplification. The resultant AAV⁃shNC and AAV⁃shCTSK were injected into the epididymal adipose tissue of mice via in situ injection. Two weeks post⁃injection , the expression of CTSK protein was evaluated through immunoblotting assay and the size of intracellular lipid droplets in mouse adipose tissue was detected by HE staining. Results : Successful acquisition of AAV⁃shNC and AAV⁃shCTSK adeno⁃associated viruses was achieved. In mice subjected to in situ injection of AAV⁃shCTSK , effective knockdown of CTSK in adipose tissue was confirmed , accompanied by a significant reduction in the size of white adipocytes. Conclusion A knockdown adeno-associated virus targeting CTSK in mice is successfully constructed , and CTSK knockdown in adipose tissue leads to a notable decrease in lipid content.

Pediatric malocclusion is common in dentistry.Some children with malocclusion combined with obstruc-tive sleep apnea-hypopnea syndrome(OSAHS)often fail to receive appropriate treatment due to a lack of multidisci-plinary diagnosis and treatment.It can cause abnormal ventilation during sleep,affecting the central nervous system and cardiovascular development and even causing neurological and behavioral problems.Pediatric OSAHS is caused by the narrowing of the upper respiratory tract,characterized by specific facial bone characteristics and neuromuscular factors and correlated with malocclusion.Due to its diverse clinical manifestations and etiology,the diagnosis and treatment of pediatric OSAHS require an interdisciplinary,personalized,and specialized approach.Questionnaires and physical ex-aminations can be used for preliminary screening.Moreover,children's stomatology and otorhinolaryngology examina-tions are the basis for disease diagnosis.Polysomnography(PSG)is currently the direct diagnostic method.There are var-ious treatment methods for OSAHS in children,and for OSAHS caused by adenoid tonsil hypertrophy,adenoidectomy and tonsillectomy are the main treatments.Othodontic treatment including mandibular advancement and rapid maxillary expansion et al is also effective for OSAHS in children with malocclusion.Currently,there is limited research on the cor-relation between childhood malocclusion and OSAHS,and multidisciplinary combination therapy may improve the cure rate,but there is a lack of sufficient evidence.In the future,the pathogenesis of OSAHS should be further elucidated,and research on multidisciplinary combination therapy should be promoted to achieve early intervention and treatment for potential and existing patients.

Aim To investigate the serum levels of thrombin-antithrombin complex(TAT),tissue type plas-minogen activator-inhibitor complex(t-PAIC)and thrombomodulin(TM)in patients with intracranial atherosclerotic steno-sis(ICAS),and their correlations with the degree of stenosis.Methods A total of 196 ICAS patients(ICAS group)who underwent treatment in Cangzhou People's Hospital from January 2021 to February 2023 were enrolled as research sub-jects.Based on the degree of vascular stenosis,they were separated into three groups:mild group(n=78),moderate group(n.=64),and severe group(n=54).A group of 196 healthy outpatient with similar clinical basic data to ICAS patients was selected as controls.The serum levels of TAT,t-PAIC,and TM in each group were compared;Spearman method was applied to analyze the correlation between serum levels of TAT,t-PAIC,TM and stenosis severity in ICAS pa-tients;Multivariate Logistic regression was applied to analyze the influencing factors of severe stenosis in ICAS patients;ROC curve was applied to analyze the predictive value of serum TAT,t-PAIC,TM and total cholesterol(TC)levels for se-vere stenosis in ICAS patients.Results Compared with the control group,the serum levels of TAT,t-PAIC,and TM were significantly increased in the ICAS group(P＜0.05);the levels of serum TAT,t-PAIC,TM,and TC in the mild,moderate,and severe groups increased accordingly(P＜0.05).Spearman analysis showed that the serum levels of TAT,t-PAIC,and TM in ICAS patients were positively correlated with the degree of stenosis(r=0.574,0.695,0.628;all P＜0.05).Multivariate Logistic regression analysis showed that TAT,t-PAIC,TM,and TC were independent risk factors for severe stenosis in ICAS patients(P＜0.05).The ROC curve showed that the AUC of severe stenosis in ICAS patients predicted by combination of TAT,t-PAIC,TM,and TC was 0.927,with a sensitivity of 83.33％and a specificity of 86.62％,which was superior to the independent prediction of TAT,t-PAIC,TM and TC(Zcombined detection-TAT=4.617,Zcombined deteetion-t-PAIC=4.024,Zcombined detection-TM=4.004,Zcombined detection-TC=7.078,all P=0.000).Conclusion The ser-um levels of TAT,t-PAIC,and TM in the ICAS group were significantly increased,and were positively correlated with the severity of stenosis.The combination of the three and TC has a high predictive value for the occurrence of severe stenosis in ICAS patients.

Objective:To explore the effects of graft recipient weight ratio (GRWR) on the early prognosis (within 1 year after operation) of recipients of different ages after split liver transplantation (SLT) in children.Methods:From April 2015 to December 2022, the relevant clinical data were retrospectively reviewed for 188 children aged under 12 years undergoing initial SLT. Based upon operative age, they were assigned into groups of L (age≤18 months, 123 cases) and H (18 months< age≤12 years, 65 cases). Draw receiver operating characteristic (ROC) curves for predicting survival rates in H and L groups using GRWR and determine the cut-off value, and subgroup dassification was based the value. Compare the general condition, intraoperative condition, postoperative condition, and major complications of recipients. Follow-ups were conducted until 12 months post-SLT, death or retransplantation within 12months post-SLT. Kaplan-Meier survival rate analysis was utilized for comparing early postoperative survival rate of recipient/graft. The incidence of major early postoperative complications was examined by χ2 test or Fisher exact probability method. Results:The survival rate of recipients at Month 12 post-SLT was 92.6% (174/188), and graft survival rate was 91.0% (171/188). The survival rate of recipients in group L at Month 12 post-SLT was 94.3% (116/123), and graft survival rate was 92.7% (114/123). The GRWR value determined of 3.1 %. According to the level of GRWR, group L was divided into groups of L-L (GRWR≤3.1%, 36 cases) and L-H (GRWR>3.1%, 87 cases) while group H groups of H-L (GRWR≤3.1%, 55 cases) and H-H (GRWR>3.1%, 10 cases). The survival rates of recipients in groups L-L/L-H were 88.9% (32/36) and 96.6% (84/87) at Month 12 post-SLT. Inter-group difference was not statistically significant ( P=0.077). Graft survival rates were 83.3% (30/36) and 96.6% (84/87 ). Inter-group difference was statistically significant ( P=0.007). The intraoperative cold ischemia time were 479.0 (194.0, 593.0) min and 204.0 (122.0, 495.0) min in groups L-L/L-H. Inter-group difference was statistically significant ( P=0.002 ). The incidence of hepatic artery thrombosis were 13.9 % (5/36) and 2.3 % (2/87) in groups L-L/L-H. Inter-group difference was statistically significant ( P=0.036). The survival rate of recipients in group H at Month 12 post-SLT was 89.2% (58/65), and graft survival rate was 87.7% (57/65). No significant inter-group difference existed during surgery ( P>0.05 ). The survival rates of recipients in group H-L/H-H at Month 12 post-SLT were 92.7 % (51 /55) and 70.0 % (7/10 ). Inter-group difference was statistically significant ( P=0.019). Graft survival rates were 90.9% (50/55) and 70.0% (7/10). Inter-group difference was statistically significant ( P=0.036). No significant inter-group difference existed in the incidence of complications ( P>0.05) . Conclusion:During pediatric SLT, recipients of different ages have different requirements for GRWR. GRWR≤3.1 % implies poor early prognosis of recipients aged ≤18 months and GRWR>3.1% is associated with poor early prognosis of recipients aged between 18 months and 12 years.

Objective:To explore the risk factors and treatments of portal vein thrombosis (PVT) in children after pediatric living donor liver transplantation (pLDLT) .Method:From January 2014 to December 2021, the relevant clinical data were retrospectively reviewed for 975 LDLT children at Department of Pediatric Organ Transplantation of Tianjin First Central Hospital. Based upon the postoperative occurrence of PVT, they were assigned into two groups of PVT (19 cases) and non-PVT (956 cases). Univariate and multivariate analyses were performed for screening the risk factors of PVT post-LDLT and discussing the managements and prognoses of PVT.Result:Among them, overall incidence of PVT post-LDLT was 1.9% (19/975), and median time for an initial occurrence of PVT 8 (1-495) day. Single-factor analysis indicated that donor height ( P=0.014), operative duration ( P=0.002) and vascular interposition ( P=0.001) were correlated with the occurrence of postoperative PVT post-pLDLT. Multifactorial analysis revealed that operative duration ( P=0.008) and vascular interposition ( P<0.01) were independent risk factors for PVT post-pLDLT. For 19 cases of postoperative PVT, the measures included surgical thrombectomy (8 cases), urokinase thrombolysis plus warfarin anticoagulation (3 cases), interventional treatment (3 cases), warfarin anticoagulation (4 cases) and retransplantation (1 cases). After treatment, the outcomes were a disappearance of PVT (15 cases), symptomatic improvement (2 cases) and unrelated mortality (2 cases) . Conclusion:During pLDLT, intraoperative placement of blood vessels and operative duration are independent risk factors for the occurrence of PVT. Timely standardized treatment may achieve satisfactory therapeutic outcomes.

Objective To investigate the effects of fentanyl citrate on pain in aged rats with hip fracture and its underlying mechanisms. Methods Thirty aged rats with hip fracture were divided into model group (n=10), fentanyl citrate group (n=10), and fentanyl citrate + Sar-SP group (n=10). Additionally, healthy rats from the same period were included as normal group (n=10).Rats in the fentanyl citrate group received intravenous tail injection of 10 μg/kg fentanyl citrate, while those in the fentanyl citrate + Sar-SP group underwent intrathecal injection of 10 μL Sar-SP for fentanyl citrate intervention. Rats in the normal and model groups received intravenous tail injection of an equal volume of saline. Pain threshold, weight-bearing capacity, temperature, and dorsoventral thickness of the hind paws were measured before treatment (0 h) and at 1, 6, 24, and 168 h post-treatment. Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and glutamic acid (Glu) in the spinal cord tissue of rats. Immunohistochemical staining was employed to observe the positive expression rates of ionized calcium-binding adapter molecule 1 (Iba-1), glial fibrillary acidic protein (GFAP), and substance P (SP) in the spinal cord tissue. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was conducted to detect the relative expression levels of SP and tachykinin receptor 1 (TACR1) mRNA in the hippocampus tissue of rats. Western blot analysis was performed to assess the relative expression levels of vimentin (VIM), nuclear receptor corepressor 1 (NOCR), ciliary neurotrophic factor receptor (CNTFR), and epidermal growth factor receptor (EGFR) proteins in the spinal cord tissue. Results Compared with the normal group, rats in the model group exhibited decreased pain threshold and weight-bearing capacity, increased hind paw temperature and dorsoventral thickness, as well as elevated levels of TNF-α, IL-6, IL-1β, and Glu, increased positive expression rates of Iba-1 and GFAP, heightened positive expression rate and relative mRNA expression levels of SP and TACR1, and augmented relative protein expression levels of VIM, NOCR, CNTFR, and EGFR in the spinal cord tissue at 0 h before treatment and at 1, 6, 24, and 168 h post-treatment (P < 0.05). Compared with the model group, rats in the fentanyl citrate group showed increased pain threshold and weight-bearing capacity, decreased hind paw temperature and dorsoventral thickness, as well as reduced levels of TNF-α, IL-6, IL-1β, and Glu, decreased positive expression rates of Iba-1 and GFAP, lowered positive expression rate and relative mRNA expression levels of SP and TACR1, and decreased relative protein expression levels of VIM, NOCR, CNTFR, and EGFR in the spinal cord tissue at 1, 6, 24, and 168 h post-treatment(P < 0.05). Compared with the fentanyl citrate group, rats in the fentanyl citrate+Sar-SP group demonstrated decreased pain threshold and weight-bearing capacity, increased hind paw temperature and dorsoventral thickness, elevated levels of TNF-α, IL-6, IL-1β, and Glu, increased positive expression rates of Iba-1 and GFAP, heightened positive expression rate and relative mRNA expression levels of SP and TACR1, and elevated relative protein expression levels of VIM, NOCR, CNTFR, and EGFR in the spinal cord tissue at 1, 6, 24, and 168 h post-treatment(P < 0.05). Conclusion The improvement in pain in aged rats with hip fracture by fentanyl citrate may be associated with the reduction of SP expression in the spinal cord that inhibits glial cell activation and exhibits anti-inflammatory effects.

Protein arginine methylation is an important post-translational modification (PTM) mediated by protein arginine methyltransferase (PRMTs), which is closely related to the occurrence and development of many diseases. Methylation of arginine is closely related to inflammatory diseases and fracture healing. Decreased expression of PRMTs can lead to delayed or even non-healing of fractures. Both PRMT5 and PRMT6 play an important role in fracture healing and are closely related to the PI3K-AKT and NF-κB pathways. Exploration the relationship between protein arginine methylation and fracture healing can provide a new way to prevent delayed or even non-healing fracture.

Objective To explore the important factors influencing organ donation willingness and coordination effect of organ donation coordinators. Methods A questionnaire survey was conducted among 349 national organ donation coordinators by convenience sampling, including 145 males and 204 females, aged 27 (23, 36) years. Multiple linear regression and disordered logistic regression were used to investigate the important factors influencing the willingness to donate organs and coordination effects. Results Among 349 organ donation coordinators, 146 (41.8%) were willing to donate organs, including 101 (28.9%) who had signed the consent card for organ donation. Adequate awareness of organ donation laws, high education level, marital experience, and good self-perceived health status all showed positive effects on organ donation willingness of organ donation coordinators (all P < 0.05). High income, long length of service as organ donation coordinators, full-time mode of employment, high willingness to donate organs, and adequate awareness of donation conditions and donation procedures all showed positive effects on the coordination effect of organ donation coordinators (all P < 0.05). Conclusions The willingness to donate organs is increased as the higher awareness of organ donation laws of organ donation coordinators, while enhancing the willingness to donate organs of organ donation coordinators exerts positive impact upon improving the coordination effect of organ donation coordination. Therefore, an all-round organ donation coordinator training system should be established to improve the success rate of organ donation advocacy and promote the development of organ donation.

Objective:To investigate the changes of hippocampal gray matter volume and expression of candidate immune related genes in a rat model of schizophrenia established by repeated administration of dizocilpine(MK-801).Methods:Thirty SPF grade Sprague-Dawley male rats at postnatal day 28 were randomly divided into MK-801 medium-dose (0.25 mg/kg) group, MK-801 high-dose(0.50 mg/kg) group and normal saline (5 mL/kg) group according to random number table method, with 10 in each group.Rats were given continuous intraperitoneal administration according to grouping once a day for 14 days.Open field test, novel object recognition test and Y-maze test were used at postnatal day 60 to detect spontaneous activity, exploration ability, anxiety level, object recognition memory ability and spatial working memory of rats, respectively.At postnatal day 67, structural magnetic resonance imaging was used to detect the changes of hippocampal gray matter volume in rat.And at postnatal day 70, qRT-PCR was used to detect the expression of candidate immune-related genes in rat hippocampus.SPSS 25.0 was used for statistical analysis, one-way ANOVA was used for comparison among multiple groups, and Tukey test was used for further pairwise comparisons.Results:(1)The behavioral results showed that there were significant differences in the total movement distance, central area activity time, novel object recognition index, and spontaneous correct alternation rate among the three groups ( F=11.15, 10.11, 13.62, 11.99, all P<0.05). The total movement distances in MK-801 medium-dose group and MK-801 high-dose group ((21.44±2.17) m, (22.87±1.96)m) were higher than that in the normal saline group ((18.70±1.88) m) (both P<0.05). The activity time of the central area in the MK-801 medium-dose group and MK-801 high-dose group((3.24±1.58) s, (2.50±1.32) s) were lower than that of the normal saline group ((6.05±2.48)s) (both P<0.01). Novel object recognition indexes in the MK-801 medium-dose group and MK-801 high-dose group((56.10±3.99)%, (54.00±6.41)%) were both lower than that in the normal saline group ((65.90±5.65)%)(both P<0.01), and the rates of spontaneous correct alternation ((54.60±7.03)%, (51.60±8.84)%) in the two groups were lower than that of the normal saline group ((68.40±8.57)%) (both P<0.01). (2) The results of structural magnetic resonance imaging showed that there were significant differences in the volume of hippocampal gray matter among the three groups ( F=9.24, P<0.001). The volumes of hippocampal gray matter in MK-801 medium-dose group and MK-801 high-dose group were lower than that in normal saline group(both P<0.001). (3)By constructing protein-protein interaction network, four candidate immune related genes were screened out: neuropeptide Y (NPY), somatostatin (SST), cholecystokinin (CCK) and tachykinin 1 (TAC1). The results showed that the mRNA expression levels of NPY, SST and CCK in the hippocampus of the three groups were significantly different ( F=11.41, 10.43, 5.85, all P<0.05), but there was no statistical difference in the TAC1 mRNA expression level ( F=0.08, P>0.05). The mRNA levels of NPY, SST and CCK in the hippocampus of rats in the MK-801 high-dose group were lower than those in the normal saline group (all P<0.05). Conclusion:Both medium dose and high dose MK-801 administration can reduce the volume of hippocampal gray matter in schizophrenia model rats, but they have different effects on the expression of hippocampal immune related genes, of which high dose administration has a greater effect.

Geriatric depression in the elderly is becoming one of the most common psychiatric disorders affecting older people's mental and physical health.However, there is currently no systematic review on animal models for geriatric depression.Therefore, this paper analyzes and summarizes the animal models commonly used in geriatric depression studies and the application of antidepressants in geriatric depression models based on relevant national and international literature of recent years, aiming to provide insights on research approaches and considerations on study methods for geriatric depression.