Background Job burnout has become an important factor affecting the mental and physical health and work efficiency of college counselors, and indirectly affects the quality and development of talent cultivation for college students. Objective To explore the relationship between job stress, job crafting, and job burnout among college counselors, and to test the mediating role of job crafting between job stress and job burnout, in order to take targeted measures to alleviate job stress and job burnout of college counselors, reduce associated health risks, and improve the effectiveness of higher education. Methods An anonymous questionnaire survey was conducted among 400 counselors from social network communication groups by convenience sampling. The Counselor Work Stress Scale, Job Crafting Scale, and Maslach Burnout Inventory-General Survey were used. Harman's single-factor method was used to evaluate common method bias in the survey data. One-way ANOVA was applied to test the difference in job stress, job crafting, and job burnout among college counselors by demographic characteristics, and chi-square test was used to analyze the difference in reporting job burnout. Partial correlation analysis was used to evaluate the correlation between selected variables. Structural equation modeling was used to analyze the relationship of job stress, job crafting, and job burnout among college counselors, and Bootstrap analysis was used to test if there was a mediating effect of job crafting on the relationship between job stress and job burnout. Results Of the 390 questionnaires recovered, there were 338 valid questionnaires (86.67%). Among the included subjects, the mean scores of job stress, job crafting, and job burnout were (2.70±0.62), (3.77±0.62), and (2.09±1.09), respectively. The positive rate of job burnout was 76.9% (260/338), with a positive rate of 72.8% in exhaustion dimension and 59.8% in cynicism dimension. There were significant differences in job crafting scores among the college counselors by different genders and professional titles (P<0.05). Female counselors had significantly higher job burnout scores and positive rates than male counselors (P<0.05). The partial correlation analysis showed that job stress, work load, school evaluation and expectation, and interpersonal relationship were positively correlated with job burnout (r=0.562, 0.442, 0.473, and 0.455, respectively, P<0.01), and negatively correlated with job crafting (r=−0.271, −0.169, −0.246, and −0.247, respectively, P<0.01); job crafting, cognitive crafting, relationship crafting, and task crafting were negatively correlated with job burnout (r=−0.447, −0.452, −0.366, and −0.340, respectively, P<0.01). The modified structural equation modeling indicated that job stress negatively affected job crafting (b=−0.348, P<0.001) and positively affected job burnout (b=0.454, P<0.001); job crafting negatively affected job burnout (b=−0.459, P<0.001), and played a partial mediating role in the relationship between job stress and job burnout, and the effect value was 0.160 (95%CI: 0.102, 0.230) that accounted for 26.10% of the total effect. Conclusion Job burnout among the college counselors is prominent. Job crafting presents an inhibitory effect on job burnout. Job stress indirectly affects the occurrence of job burnout by inhibiting the generation of job crafting.

Objective To investigate the role of G protein signal regulator 2(RGS2)in regulating the formation of angiotensin Ⅱ(Ang Ⅱ)-induced aortic dissection.Methods C57BL/6 mice were divided into 3 groups:control group(n=10),Ang Ⅱ group(n=20),Ang Ⅱ+sh-RGS2 group(n=20).The Ang Ⅱ group and Ang Ⅱ+sh-RGS2 group established an aortic dissection model.The incidence of aortic dissection was evaluated in vivo,and the phenotypic transformation of VSMC was evaluated in vitro and in vivo.Results Knockdown of RGS2 largely coun-teracted Ang Ⅱ-induced inhibition of αSMA,ACTA2 and MYH11,and suppressed Ang Ⅱ-induced SPP1 and Vim-entin in VSMC.The incidence of aortic dissection in Ang Ⅱ group and Ang Ⅱ+sh-RGS2 group were 45％(9/20)and 10％(2/20),respectively.Fewer elastic lamina thickening,aortic rupture,and aortic wall collagen fiber con-tent were observed in Ang Ⅱ+sh-RGS2 group compared to Ang Ⅱ group.In addition,compared with the Ang Ⅱgroup,the maximum diameter of the aorta in the Ang Ⅱ+sh-RGS2 group was significantly reduced(P＜0.05).In addition,the ACTA2 and MYH11 proteins in the aorta of the Ang Ⅱ+sh-RGS2 group were significantly higher than those in the Ang Ⅱ group(P＜0.01),while the RGS2,SPP1,and Vimentin proteins significantly decreased(P＜0.01).Conclusion Knockdown of RGS2 inhibits the transformation of Ang Ⅱ-induced VSMC from a contractile phenotype to a synthetic phenotype,thereby reducing the incidence of aortic dissection formation.

Objective:To investigate the diagnosis, treatment, clinical characteristics and potential high-risk factors of cerebral venous sinus thrombosis (CVST) during the treatment of acute lymphoblastic leukemia (ALL) with pegaspargase.Methods:The medical history, diagnosis and treatment process, laboratory examination and imaging examination results of 3 ALL patients with CVST during pegaspargase treatment in the First Affiliated Hospital of Suzhou University in March and November 2021 and September 2022 were retrospectively analyzed, and the relevant literature was reviewed.Results:Three patients were all female, with the aged between 15 and 35 years old, including 2 cases of B-ALL and 1 case of T-ALL. All patients developed nervous system symptoms after pegaspargase chemotherapy, and were diagnosed as CVST by imaging examination. During the pegaspargase treatment, 2 patients took norethisterone, and 1 patient underwent induced labor and curettage. The levels of sexual hormones in the 3 patients had non-physiological changes. The main CVST lesions were located in the superior sagittal sinus, transverse sinus and sigmoid sinus. One patient had cerebral hemorrhage at the same time. When thrombus occurred, the fibrinogen (Fib), antithrombin Ⅲ (AT Ⅲ) activity, protein C activity and protein S activity of the patients were significantly lower than those before, D-dimer was significantly higher, and lupus anticoagulant and anticardiolipin antibody were negative. The thrombosis treatment was mainly anticoagulation, and 1 patient underwent thrombolysis. Two patients had no sequelae of nervous system, and 1 patient had the sequelae of muscle weakness.Conclusions:Patients with ALL should be alert to the occurrence of CVST when they have nervous system symptoms during pegaspargase chemotherapy. The diagnosis of CVST mainly depends on cranial imaging. Anticoagulation is the main thrombosis treatment, thrombolysis and interventional thrombectomy are feasible for some patients, with few neurological sequelae. The use of second-generation progesterone drugs and the non-physiological fluctuation of sex hormones may be the potential risk factors of CVST.

ObjectiveTo reveal the effects of different microbial agents on quality of Lycii Fructus by comparing the differences in the contents of multiple types of chemical components in Lycii Fructus after the application of different microbial agents. MethodTaking Ningqi No. 7 as experimental material， four microbial agents， namely Peiyuan combined with Xinterui（TP group）， Trichoderma harzianum combined with Bacillus subtilis（BW group）， Genwuyou（MT group） and Junyiduo（JYD group）， were applied， and no microbial agents was used as the blank group（CK group）. Then the contents of total phenolics， total flavonoids， saccharides， amino acids， nucleosides and bases， betaine and other components in Lycii Fructus were determined by ultraviolet spectrophotometry（UV）， high performance liquid chromatography（HPLC） and ultra-high performance liquid chromatography-mass spectrometry（UHPLC-MS/MS）， and the methods such as multiple comparisons， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） were used to analyze the effect of different microbial agents on the quality of Lycii Fructus. ResultMicrobial agents had different effects on chemical components of Lycii Fructus. The content of total phenolics was the highest in the TP group， and it varied significantly from the CK group（P<0.05）. The total flavonoid content was the highest in the BW group， followed by the TP group. Both polysaccharide and alduronic acid contents were the highest in the JYD group. Betaine content in the TP and BW groups were significantly higher than that in the CK group（P<0.05）. For the determined 23 kinds of amino acids， most of them were the lowest in the JYD group， and the highest in the MT group， while the nucleoside bases were higher in the MT and BW groups. It indicated that Lycii Fructus from different treatment groups could be distinguished clearly based on the determined 45 chemical components. The result of PLS-DA showed that the major differential components in each group were polysaccharides， glucose， fructose， betaine， alduronic acid， asparagine， sucrose， threonine， total flavonoids， alanine and total phenolics. The results of PCA composite scores based on the main differential components showed that composite scores of chemical components in each group were BW group>TP group>MT group>CK group>JYD group. ConclusionThe application of microbial agents of BW， TP and MT can promote the quality improvement of Lycii Fructus， and the application of JYD can promote the accumulation of polysaccharides and alduronic acid to a certain extent， but the overall effect on the quality of Lycii Fructus is not clear. This study lays the foundation for the green and healthy development of Lycii Fructus industry.

Objective: Depression was common during coronavirus disease-2019 (COVID-19) pandemic, while the association of perceived stress with depression among vaccinated healthcare workers has not been investigated. This study aimed to address this issue. Methods: We included a total of 898 fully vaccinated healthcare workers during the outbreak of severe acute respiratory syndrome coronavirus 2 Delta variant in Nanjing, 2021. Depression was ascertained by Patient Health Questionnaire-9, with a cut-off score of ≥5 indicative of mild-to-severe depression. Perceived stress, resilience, and compassion fatigue were assessed by Perceived Stress Scale-10, Resilience Scale-25, and Professional Quality of Life Scale version-5, respectively. Logistic regression analyses were used to estimate the odds ratio (OR) and 95% confidence interval (CI), along with subgroup and mediation analyses. Results: The prevalence of mild-to-severe depression was 41.1% in vaccinated healthcare workers. The odd of mild-to-severe depression was increased with higher perceived stress. Compared with vaccinated healthcare workers with the lowest tertile of perceived stress, those with the highest tertile had increased odds of mild-to-severe depression by 120% (OR 2.20, 95% CI 1.46 to 3.31) after multivariable-adjustment. However, perceived stress was not associated with mild-to-severe depression in vaccinated healthcare workers with strong resilience, but was in those with weak resilience (pinteraction=0.004). Further analysis showed that compassion fatigue mediated the relationship between perceived stress and mild-to-severe depression, with a mediating effect of 49.7%. Conclusion Perceived stress was related to an increased odd of mild-to-severe depression in vaccinated healthcare workers during COVID-19 pandemic, and this relationship might be explained by compassion fatigue.

Objective: To investigate the antimicrobial resistance in and multilocus sequence typing (MLST) of Campylobacter jejuni in meat and meat products in Liaoning Province, so as to provide insights into the management of C. jejuni infection. Methods: Nine C. jejuni isolates from meat and meat products in 2020 were subjected to whole genome sequencing, and the minimum inhibitory concentration was measured with the agar dilution method. MLST of C. jejuni isolates was performed with the microbial resistance mechanism traceability reference database and analysis system. Results: Six drug-resistant C. jejuni isolates were detected, and there were four multidrug-resistant isolates. There were six C. jejuni isolates resistant to tetracycline, five isolates resistant to nalidixic acid, four isolates resistant to ciprofloxacin, two isolates resistant to florfenicol, one isolate resistant to gentamicin and one isolate resistant to streptomycin. Nine C. jejuni isolates showed sensitive to azithromycin, chloramphenicol and clindamycin. MLST identified six ST types in nine C. jejuni isolates, with ST45 and ST2274 as the predominant type, and detected one isolate with unclassified ST type. Phylogenetic analysis showed that KW028 and KW029 of ST45 type were closely related and had high homology, and KW040 and KW042 of ST2274 type were closely related with high homology, while KW007 of ST6701 type was closely related to KW040 and KW042 of ST2274 type, with only one pgm housekeeper gene in difference. Conclusions High resistance to tetracycline, nalidixic acid and ciprofloxacin was detected in nine C. jejuni isolates from meat and meat products, and ST45 and ST2274 were predominant ST types of C. jejuni.

Objective:To investigate the effect and regulation of umbilical cord-derived mesenchymal stem cells (UC-MSCs) on islets function and NOD-like receptor family, pyrin domain containing 3 (NLRP3) and autophagy in type 2 diabetic mellitus (T2DM) mice.Methods:Experimental study. Twenty, 8-week-old, male C57BL/6J mice were selected and divided into a normal control group ( n=5) and a high-fat feeding modeling group ( n=15). The model of T2DM was established by high-fat feeding combined with intraperitoneal injection of low-dose streptozotocin. After successful modeling, those mice were divided into a diabetes group ( n=7) and a UC-MSCs treatment group ( n=7). The UC-MSCs treatment group was given UC-MSCs (1×10 6/0.2 ml phosphate buffer solution) by tail vein infusion once a week for a total of 4 weeks; the diabetes group was injected with the same amount of normal saline, and the normal control group was not treated. One week after the treatment, mice underwent intraperitoneal glucose tolerance tests and intraperitoneal insulin tolerance tests, and then the mice were sacrificed to obtain pancreatic tissue to detect the expressions of interleukin-1β (IL-1β) and pancreatic and duodenal homeobox 1 (PDX-1) by immunofluorescence. The bone marrow-derived macrophages were stimulated with lipopolysaccharide and adenosine triphosphate (experimental group) in vitro, then co-cultured with UC-MSCs for 24 h (treatment group). After the culture, enzyme-linked immunosorbent assay was used to detect the secretion level of IL-1β in the supernatant, and immunofluorescence staining was used to detect the expression of NLRP3 inflammasome, and related autophagy proteins. Statistical analysis was performed using unpaired one-way analysis of variance, repeated measure analysis of variance. Results:In vivo experiments showed that compared with the diabetes group, the UC-MSCs treatment group partially repaired islet structure, improved glucose tolerance and insulin sensitivity (all P<0.05), and the expression of PDX-1 increased and IL-1β decreased in islets under confocal microscopy. In vitro experiments showed that compared with the experimental group, the level of IL-1β secreted by macrophages in the treatment group was decreased [(85.9±74.6) pg/ml vs. (883.4±446.2) pg/ml, P=0.001], the expression of NLRP3 inflammasome and autophagy-related protein P62 was decreased, and the expressions of microtubule-associated protein 1 light chain 3β (LC3) and autophagy effector Beclin-1 were increased under confocal microscopy. Conclusions:UC-MSCs can reduce the level of pancreatic inflammation in T2DM mice, preserving pancreatic function. This might be associated with the ability of UC-MSCs to inhibit the activity of NLRP3 inflammasomes in macrophages and enhance autophagy levels.

Objective:To investigate the changes of various cytokines and coagulation function in B cell acute lymphoblastic leukemia(ALL) patients with different CRS scores during CD19-CAR-T cell immunotherapy.Methods:87 patients with B-ALL hospitalized in the First Affiliated Hospital of Soochow University and 30 normal controls were enrolled into this study from July 2018 to October 2020. The age of the patients was 32(20, 56) years old and 36(41.4%) were female. All these coagulation indicators, prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimer, fibrinogen (Fg) were analyzed by automatic blood coagulation in B-ALL patients before and after treated with CAR-T cell. The ratio of CD19-CAR-T cells and the expression of IL-6, IL-10, IFN-γ, TFN-α, IL-2, IL-4, and IL-17A were analyzed using flow cytometry. The patients′ clinical parameters were detected, and the CRS classification of severity was made according to the standard of consensus.Results:Patients with CRS>3 had prolonged PT and APTT, increased D-dimer, and decreased fibrinogen ( P<0.05). The levels of cytokines of IFN-γ, IL-6, and IL-10 were significantly higher in patients with CRS>3 than that in controls ( P<0.05).The D-dimer level is positively correlated with IL-10. Conclusion:Patients with severe CRS grading have significant coagulation dysfunction in CD19-CAR-T cell immunotherapy. Cytokines IFN-γ, IL-6, and IL-10 may affect coagulation function and CRS grading during CD19-CAR-T cell immunotherapy.

Streptococcus pyogenes Cas9 (SpCas9) has become a powerful genome editing tool, but has a limited range of recognizable protospacer adjacent motifs (PAMs) and shows off-target effects. To address these issues, we present a rational approach to optimize the xCas9 mutant derived from SpCas9 by directed evolution. Firstly, energy minimization with the Rosetta program was applied to optimize the three-dimensional structure of Cas9 to obtain the lowest energy conformation. Subsequently, combinatorial mutations were designed based on the mutations sites of xCas9 acquired during the directed evolution. Finally, optimal mutants were selected from the designed mutants by free energy ranking and subjected to experimental verification. A new mutant yCas9 (262A/324R/409N/480K/543D/694L/1219T) with multiple PAM recognition ability and low off-target effects was obtained and verified by DNA cleavage experiments. This mutant recognizes the NG, GAA and GAT PAMs and shows low off-target DNA cleavage activity guided by mismatched sgRNA, thus provides a gene editing tool with potential applications in biomedical field. Furthermore, we performed molecular dynamics simulations on the structures of SpCas9, xCas9 and yCas9 to reveal the mechanisms of their PAM recognition and off-target effects. These may provide theoretical guidance for further optimization and modification of CRISPR/Cas9 proteins.
CRISPR-Associated Protein 9/metabolism* ; CRISPR-Cas Systems/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats ; Gene Editing ; RNA, Guide/genetics* ; Streptococcus pyogenes/metabolism*

Objective:To analyze the correlation between 21-gene recurrence score (RS) and clinicopathological characteristics of patients with Lumina type breast cancer, and to explore its significance in individualized treatment.Methods:The clinicopathological data of 59 patients with surgical resection and pathological diagnosis of Lumina type breast cancer in Shanxi Provincial Cancer Hospital from May 2018 to May 2019 were retrospectively analyzed. Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of 21 gene and RS was calculated. According to the 21-gene RS, the patients were divided into low recurrence risk group (RS < 18 points), intermediate recurrence risk group (RS 18-31 points) and high recurrence risk group (RS > 31 points). Univariate and multivariate logistic regression analyses were made to evaluate the correlations between different recurrence risk and clinicopathological characteristics of patients and their influence on the choice of adjuvant chemotherapy.Results:Based on the 21-gene RS, 29 patients were in low recurrence risk group, 22 cases were in intermediate recurrence risk group, and 8 cases were in high recurrence risk group. Single-factor analysis showed that age ( P = 0.012), maximum mass diameter ( P = 0.031), histological grade ( P = 0.036), progesterone receptor (PR) level ( P = 0.015), Ki-67 positive index ( P = 0.049) and molecular typing ( P = 0.010) were influencing factors of 21-gene RS recurrence risk. Multivariate logistic regression analysis showed that the age and Ki-67 positive index were negatively correlated with 21-gene RS recurrence risk (both P < 0.05). After grouping according to the 21-gene RS, 17 patients in the intermediate recurrence risk group (according to the traditional postoperative recurrence risk grouping method for breast cancer) were classified as low recurrence risk group, and 4 patients in the low recurrence risk group were classified as intermediate recurrence risk group ( χ2 = 4.535, P = 0.033). After grouping based on 21-gene RS, the number of patients who needed chemotherapy in individualized treatment decreased. Of the 17 cases, 11 cases did not undergo postoperative chemotherapy, and the remaining patients received chemotherapy. The postoperative follow-up period was 11-22 months. As of March 2020, there was no recurrence or disease progress. Conclusion:The 21-gene RS can provide objective basis for the individualized precise treatment and prognosis prediction for patients with early-stage Lumina type breast cancer.