The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Objective:To examine the association among childhood emotional neglect, thwarted belongingness, perceived burdensomeness, and depression in junior high school students.Methods:From October to December 2024, a total of 314 junior high school students were recruited in Dongying City, Shandong Province. The Chinese versions of the childhood trauma questionnaire-short form (CTQ-SF), the center for epidemiologic studies depression scale (CES-D), and the interpersonal needs questionnaire (INQ) were administered for investigation. SPSS 27.0 software was used for common-method bias test, descriptive analysis, difference test, and correlation analysis. Mplus 8.3 software was employed to build a structural equation model, and the Bootstrap method was adopted for mediation analysis.Results:The scores of depression, childhood emotional neglect, thwarted belongingness, and perceived burdensomeness in junior high school students were 14.00 (9.00, 24.00), 9.00 (5.00, 12.00), 13.00 (8.00, 18.00) and 8.00 (6.00, 19.25), respectively. All variables were positively correlated with one another ( r=0.33-0.51, P<0.01). The total effect of childhood emotional neglect on depression was 0.493 (95% CI=0.396-0.591). The direct effect was 0.126 (95% CI=0.013-0.240), accounting for 25.56%(0.126/0.493) of the total effect. The total indirect effect was 0.367 (95% CI=0.287-0.447), accounting for 74.44%(0.367/0.493) of the total effect. Specifically, the pathway " childhood emotional neglect → thwarted belongingness→ depression" yielded an effect of 0.109, and the pathway " childhood emotional neglect → perceived burdensomeness → depression" yielded an effect of 0.258. Conclusion:Thwarted belongingness and perceived burdensomeness play parallel mediating effects between childhood emotional neglect and depression in junior high school students.

Objective:To examine the association among childhood emotional neglect, thwarted belongingness, perceived burdensomeness, and depression in junior high school students.Methods:From October to December 2024, a total of 314 junior high school students were recruited in Dongying City, Shandong Province. The Chinese versions of the childhood trauma questionnaire-short form (CTQ-SF), the center for epidemiologic studies depression scale (CES-D), and the interpersonal needs questionnaire (INQ) were administered for investigation. SPSS 27.0 software was used for common-method bias test, descriptive analysis, difference test, and correlation analysis. Mplus 8.3 software was employed to build a structural equation model, and the Bootstrap method was adopted for mediation analysis.Results:The scores of depression, childhood emotional neglect, thwarted belongingness, and perceived burdensomeness in junior high school students were 14.00 (9.00, 24.00), 9.00 (5.00, 12.00), 13.00 (8.00, 18.00) and 8.00 (6.00, 19.25), respectively. All variables were positively correlated with one another ( r=0.33-0.51, P<0.01). The total effect of childhood emotional neglect on depression was 0.493 (95% CI=0.396-0.591). The direct effect was 0.126 (95% CI=0.013-0.240), accounting for 25.56%(0.126/0.493) of the total effect. The total indirect effect was 0.367 (95% CI=0.287-0.447), accounting for 74.44%(0.367/0.493) of the total effect. Specifically, the pathway " childhood emotional neglect → thwarted belongingness→ depression" yielded an effect of 0.109, and the pathway " childhood emotional neglect → perceived burdensomeness → depression" yielded an effect of 0.258. Conclusion:Thwarted belongingness and perceived burdensomeness play parallel mediating effects between childhood emotional neglect and depression in junior high school students.

The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Objective:To evaluate and screen the regimens of tigecycline intraventricular injection in extensively drug resistant Acinetobacter baumannii (XDRAB) intracranial infection based on Monte Carlo simulation and pharmacokinetic/pharmacodynamic (PK/PD) model.Methods:Nine patients with XDRAB intracranial infection confirmed as having susceptibility to tigecycline or polymyxin antimicrobials from January 1, 2018 to December 31, 2023 were screened from electronic medical record system in Second Affiliated Hospital of Shandong First Medical University. WHONET software was used to extract pathogen susceptibility data isolated from cerebrospinal fluid samples. Minimum inhibitory concentration (MIC) of tigecycline against XDRAB was analyzed by drug susceptibility test; different regimens for intraventricular tigecycline injection were designed based on MIC: 2 mg/12 h, 3 mg/12 h, 4 mg/12 h, 5 mg/12 h, 6 mg/12 h, and 10 mg/12 h, with drug concentration of 0.5 mg/mL or 1.0 mg/mL once a day. Target value of PK/PD index was set as ?C max/MIC≥8; Monte Carlo was used to simulate the compliance of PK/PD index of tigecycline with different MIC against XDRAB for different dosed regimens (probability of target attainment [PTA] and cumulative fraction of response [CFR]); the best regiment was selected (screening basis: PTA≥90% or CFR≥90%). Results:(1) A total of 27 strains of pathogenic bacteria from 9 patients were extracted from drug susceptibility test, in which MIC of tigecycline against XDRAB was 55.56% for 2 mg/L, 25.93% for 4 mg/L, and 18.52% for 8 mg/L. (2) When the drug concentration was 0.5 mg/mL or 1.0 mg/mL, respectively, all 6 regimens had PTA>90% at 2 mg/L MIC; 5 regimens, except for 2 mg/12 h, had PTA>90% at 4 mg/L MIC; regimens of 5 mg/12 h, 6 mg/12 h, and 10 mg/12 h could achieve PTA>90% at 8 mg/L MIC. (3) When the drug concentration was 0.5 mg/mL, regimens of 4 mg/12 h, 5 mg/12 h, 6 mg/12 h, and 10 mg/12 h could achieve CFR>90%; when the drug concentration was 1 mg/mL, regimens of 4 mg/12 h, 5 mg/12 h, 6 mg/12 h, and 10 mg/12 h could achieve CFR>92%.Conclusion:In intraventricular tigecycline injection for XDRAB intracranial infection, 2 mg/12 h regimen is available in 2 mg/L MIC, 3 mg/12 h regimen is available in 4 mg/L MIC, and 5 mg/12 h regimen is available in 8 mg/L MIC, with either 0.5 mg/mL or 1 mg/mL concentration.

Grooming,as an evolutionarily conserved repetitive behavior,is common in various animals,including humans,and serves essential functions including,but not limited to,hygiene maintenance,thermoregulation,de-arousal,stress reduction,and social behaviors.In rodents,grooming involves a patterned and sequenced structure,known as the syntactic chain with four phases that comprise repeated stereotyped movements happening in a cephalocaudal progression style,beginning from the nose to the face,to the head,and finally ending with body licking.The context-dependent occurrence of grooming behavior indicates its adaptive significance.This review briefly summarizes the neural substrates responsible for rodent grooming behavior and explores its relevance in rodent models of neuropsychiatric disorders and neurodegenerative diseases with aberrant grooming phenotypes.We further emphasize the utility of rodent grooming as a reliable measure of repetitive behavior in neuropsychiatric models,holding promise for translational psychiatry.Herein,we mainly focus on rodent self-grooming.Allogrooming(grooming being applied on one animal by its conspecifics via licking or carefully nibbling)and heterogrooming(a form of grooming behavior directing towards another animal,which occurs in other contexts,such as maternal,sexual,aggressive,or social behaviors)are not covered due to space constraints.

Objective To evaluate the safety, effectiveness and feasibility of robotic-assisted kidney transplantation (RAKT). Methods Clinical data of 16 patients who underwent kidney transplantation were collected. Among them, 8 recipients received RAKT (RAKT group) and 8 cases underwent open kidney transplantation (OKT) with the contralateral kidney from the same donor (OKT group). Perioperative status and the recovery of renal allograft function were compared between two groups. Results All patients successfully completed the surgery. In the RAKT group, no patient was converted to open surgery. The operation time in the RAKT group was longer than that in the OKT group (P=0.015). No significant differences were observed in the serum creatinine levels before surgery and upon discharge between two groups (both P>0.05). In the OKT group, one recipient developed delayed graft function (DGF), and the remaining recipients did not experience perioperative complications. No significant difference was noted in the short-term recovery of renal allograft function between two groups (P>0.05). Conclusions Postoperative recovery of the recipients in the RAKT group is equivalent to that of their counterparts in the OKT group. RAKT is a safe and effective procedure for the team expertise in kidney transplantation.

ObjectiveBased on serum pharmacochemistry and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） the transitional components in the serum of rats after intragastric administration of water extract of Alismatis Rhizoma（AR）and salt-processed Alismatis Rhizoma（SAR） were compared. MethodSD rats were randomly divided into blank group, AR group（10 g·kg^-1） and SAR group（10 g·kg^-1）, 3 rats in each group, the administration groups were given AR and SAR aqueous extracts by gavage, respectively, and the blank group was given an equal volume of drinking water by gavage once in the morning and once in the evening, for 3 consecutive days. Sixty minutes after the last administration, blood was collected from the eye orbits, and the serum samples were prepared. The serum samples were prepared on an ACQUITY UPLC BEH C18 column（2.1 mm×50 mm, 1.7 μm） with the mobile phase of acetonitrile（A）-0.1% formic acid aqueous solution（B） in a gradient elution（0-10 min, 10%-50% A; 10-27 min, 50%-95%A; 27-27.1 min, 95%-10% A; 27.1-30 min, 10%A）, the data were collected at a flow rate of 0.3 mL·min^-1 in positive ion mode with a scanning range of m/z 100-1 200. Based on the self-constructed chemical composition library of AR, the total ion flow diagrams and secondary MS fragmentation information of the aqueous extracts of AR and SAR, as well as the administered serum and the blank serum, were compared with each other by UNIFI 1.9.2, so as to deduce the possible blood-migrating constituents and their cleavage patterns in the aqueous extracts, and the response intensity ratios of each chemical component were calculated before and after processing. ResultA total of 20 components, including 5 prototypical components and 15 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of AR. And 14 components, including 5 prototypical components and 9 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of SAR. Of these, 13 components were common to both of them, including 5 prototypical components and 8 metabolites. The 5 prototypical components were 16-oxoalisol A, alisol A 24-acetate, alisol A, alisol B and alisol C. The metabolites were mainly involved in phase Ⅰ metabolism（oxidation） and phase Ⅱ metabolism（glucuronidation）. There was a big change in the intensity of response of the common components before and after salt-processing, and the response intensities of the prototypical components, 16-oxoalisol A, alisol B and alisol C, were elevated, while the type and response intensity of metabolites were generally decreased, and it was hypothesized that the metabolic rate of terpenoids might be slowed down after salt-processing of AR, so that the blood-migrating constituents could participate in the metabolism of the body more in the form of prototypes. ConclusionSalt-processing of AR may promote the absorption of prototypical components into the blood by slowing down the metabolic rate of terpenoids, which can provide support for the research on material basis of AR and SAR.

ObjectiveTo evaluate the pharmacological effect of Alismatis Rhizoma （AR） and its processed product on rats with edema of kidney Yin deficiency and explore the mechanism. MethodA total of 42 male SPF SD rats were randomized into normal group （equivalent volume of distilled water）， model group （equivalent volume of distilled water）， positive medicine Liuwei Diguangwan group （1.4 g·kg^-1）， low- and high-dose AR groups （1， 4 g·kg^-1， respectively）， and low- and high-dose salt-processed AR （SAR） groups （1， 4 g·kg^-1， respectively）， with six rats in each group. Adriamycin （tail vein injection） and thyroxine （gavage） were used to induce edema of kidney Yin deficiency in rats except the normal group. The administration lasted 4 weeks for all the groups. After the last administration， histopathological changes of rat kidneys were observed based on hematoxylin-eosin （HE） staining. Serum content of triiodothyronine （T₃）， thyroxine （T₄）， follicle-stimulating hormone （FSH）， and testosterone （T） was determined by radioimmunoassay， and serum content of creatinine （CREA）， urea （UREA），cholesterol （CHOL） and triglyceride （TG） by automatic biochemical analyser. The levels of gonadotropin-releasing hormone （GnRH）， cyclic adenosine monophosphate （cAMP）， and cyclic guanosine monophosphate （cGMP） in plasma were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of aquaporin（AQP）-1 and AQP-2 and the transcription of mRNA in kidney were measured by immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultCompared with normal group， the rats in model group showed decrease in body mass and urine volume （P<0.01）， increase in water consumption （P<0.05）， infiltration of a large number of inflammatory cells and fibrous tissue proliferation in the kidney， rise of the expression and transcript levels of T₃， T₄， cAMP/cGMP， CREA， FSH， AQP-1， and AQP-2 （P<0.01）， the contents of CHOL and TG were significantly increased （P<0.05）， and reduction in the levels of GnRH and T （P<0.01）. Body mass increased in both the low- and high- dose groups of AR and SAR compared with that in model group， with significant differences between the low-dose AR group and the low-dose SAR group （P<0.01）. Moreover， compared with model group， low- and high-dose AR and SAR insignificantly increased the urine volume of rats， reduced the inflammatory cells in kidney tissues， significantly decreased the levels of T₄， cAMP/cGMP， UREA， CREA， FSH， CHOL and TG in serum （P<0.05，P<0.01）， and elevated the level of GnRH （P<0.01）， high-dose AR， low- and high-dose SAR significantly lowered the transcription levels of AQP-1 and AQP-2 mRNA in the kidneys of rats （P<0.01）. ConclusionBoth AR and SAR alleviated the edema of kidney Yin deficiency in rats by down-regulating the expression of AQP-1 and AQP-2 and correcting the hypothalamic-pituitary-gonadal （HPG） axis disorder.

The responsive patterns of phytochrome gene family members to photoperiod and abiotic stresses were comparatively analyzed and the favorable natural variation sites of these genes were identified. This would help understand the mechanism of phytochrome gene family in photoperiod-regulated growth and development and abiotic stress response. In addition, it may facilitate the molecular marker assisted selection of key traits in foxtail millet. In this study, we used RT-PCR to clone three phytochrome genes SiPHYA, SiPHYB and SiPHYC from ultra-late maturity millet landrace variety 'Maosu'. After primary bioinformatics analysis, we studied the photoperiod control mode and the characteristics of these genes in responding to five abiotic stresses including polyethylene glycol (PEG)-simulated drought, natural drought, abscisic acid (ABA), high temperature and NaCl by fluorescence quantitative PCR. Finally, we detected the mutation sites of the three genes among 160 foxtail millet materials and performed haplotype analysis to determine the genes' functional effect. We found that the cloned cDNA sequences of gene SiPHYA, SiPHYB and SiPHYC were 3 981, 3 953 and 3 764 bp respectively, which contained complete coding regions. Gene SiPHYB and SiPHYC showed closer evolutionary relationship. Photoperiod regulated all of the three genes, but showed more profound effects on diurnal expression pattern of SiPHYB, SiPHYC than that of SiPHYA. Under short-day, when near heading, the expression levels of SiPHYA and SiPHYB were significantly lower than that under long-day, indicating their roles in suppressing heading of foxtail millet under long-day. SiPHYB and SiPHYC were responsive to PEG-simulated drought, natural drought, ABA and high temperature stresses together. SiPHYA and SiPHYB responded differently to salt stress, whereas SiPHYC did not respond to salt stress. Re-sequencing of 160 foxtail millet materials revealed that SiPHYB was highly conservative. Two missense mutations of SiPHYA, such as single nucleotide polymorphism (SNP) 7 034 522^C→T and SNP7 036 657^G→C, led to delaying heading and increasing plant height. One missense mutation of SiPHYC, such as SNP5 414 823^G→T, led to shortening heading under short-day and delaying heading under long-day, as well as increasing plant height and panicle length regardless of photo-thermal conditions. Photoperiod showed different regulatory effects on SiPHYA, SiPHYB and SiPHYC. SiPHYB and SiPHYC jointly responded to various abiotic stresses except for the salt stress. Compared with the reference genotype, mutation genotypes of SiPHYA and SiPHYC delayed heading and increased plant height and panicle length.
Gene Expression Regulation, Plant ; Photoperiod ; Phytochrome/metabolism* ; Plant Proteins/metabolism* ; Setaria Plant/metabolism* ; Stress, Physiological/genetics*