Objective: To investigate the expression levels of serum neuron specific enolase (NSE), interleukin-1β (IL-1β)and brain derived neurotrophic factor(BDNF) in adolescents with generalized anxiety disorder(GAD), so as to provide reference for the early diagnosis and evaluation of GAD in adolescents. Methods: From March 2020 to February 2023, 97 first episode adolescents with generalized anxiety disorder aged 13-18 years were selected in the study. According to the score of generalized anxiety disorder questionnaire (GAD-7) on admission, they were divided into mild moderate GAD group (58 cases) and severe GAD group (39 cases). At the same time, 90 healthy adolescents who participated in routine physical examination in the same period were selected as the control group. Variance analysis was conducted for comparison among serum NSE, IL-1β and BDNF level. LSD t was applied for further comparison, and the bivariate Pearson linear correlation analysis was carried out for the relationship among serum NSE, IL-1β and BDNF level. Results: The differences among 3 groups of subjects on NSE and IL-1β, and BDNF were of statistical significance( F =10.73, 12.80, 20.67, P <0.01), and the differences between groups were statistically significant ( P <0.05). In the control group, serum NSE level was(8.70±1.35) μg/L, IL-1β was (18.42±5.43) pg/mL, both were the lowest. The levels of NSE and IL-1β in severe GAD group were(14.21±3.25) μg/L, (26.04±5.39)pg/mL, which were the highest. The serum BDNF level in control group was (27.16±4.42) ng/mL, which was the highest; and the severe GAD group was (10.46±3.27) ng/mL, which was the lowest. The bivariate Pearson linear correlation analysis showed that the serum NSE and IL-1β levels in GAD group were negatively correlated with the serum BDNF level ( r =-0.49, -0.57); the serum NSE level was positively correlated with the IL-1β level ( r =0.40) ( P <0.05). Conclusions The serum levels of NSE and IL-1β are abnormally increased in adolescents with GAD, and the serum levels of BDNF are significantly decreased. The serum levels of NSE, IL-1β and BDNF can be used as detection markers for adolescent GAD, which is helpful for early diagnosis and disease evaluation.

Objective:To investigate the expression levels of serum KLK5, IL-17 and IL-18 in patients with rosacea and their clinical significance.Methods:A prospective study included 65 patients with rosacea from the Department of Dermatology, Sichuan Provincial People＇s Hospital, from May to October 2022. There were 8 males and 57 females with an average age of (30.9±9.8) years, including 47 cases of erythematotelangiectatic type and 18 cases of papulopustular type. Twenty healthy individuals from the same hospital＇s physical examination during the same period were included as control group, consisting of 3 males and 17 females with an average age of (33.8±10.0) years. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of serum KLK5, IL-17, and IL-18. Investigator＇s global assessment (IGA) score was used to assess the severity of papules and pustules, and clinical erythema assessment (CEA) score was used to assess the severity of persistent erythema. The expression levels of serum KLK5, IL-17, and IL-18 were compared between the two groups, and the correlation between the severity of the disease and the expression levels of KLK5, IL-17, and IL-18 was analyzed. Additionally, the expression levels of KLK5, IL-17, and IL-18 in different subtypes of rosacea were compared.Results:The expression levels of serum KLK5, IL-17, and IL-18 in the rosacea group were (3134.01±448.82), (154.78±37.64), (236.24±38.81) pg/ml, respectively, all of which were higher than those in the healthy control group (2421.54±340.07), (95.36±24.94), (181.61±23.88) pg/ml, with statistically significant differences (all P<0.05). The levels of serum KLK5, IL-17, and IL-18 in patients with rosacea were positively correlated with IGA scores ( r=0.534, 0.767, 0.417) and CEA scores ( r=0.852, 0.459, 0.309) (all P<0.05). There were no statistically significant differences in the expression levels of serum KLK5, IL-17, and IL-18 between the erythematotelangiectatic and papulopustular subtypes of rosacea (all P>0.05). Conclusions:The expression levels of serum KLK5, IL-17 and IL-18 in patients with rosacea are higher than those in the healthy control group and are positively correlated with the severity of rosacea.

Objective To explore the current situation and influencing factors of the resilience and burnout of medical staff in a tertiary hospital in Shanghai.Methods A total of 189 medical staff in a tertiary hospital in Shanghai were surveyed by general information questionnaire,Maslach Burnout Inventory-General Survey(MBI-GS),and medical staff resilience evaluation scale.Results The total score of resilience of health care workers was 70.32±10.98,which were the scores of decision-making response,interpersonal connection,and rational thinking of medical staff in this study.These were lower than the scores of the domestic norm(all P<0.001),the flexible self-adaptation score was slightly lower than the scores of the domestic norm(P<0.05).The total burnout score was 53.89±1 5.42.The scores of emotional exhaustion and personal fulfillment of medical staff in this study were lower than the scores of the domestic norm(P<0.05),the scores of depersonalization were significantly higher than the scores of the domestic norm(P<0.001).Burnout was inversely correlated with the level of resilience.There were statistically significant differences in the levels of burnout and resilience among medical staff in terms of working years and income satisfaction(P<0.05).Income satisfaction was the influencing factor of resilience level and burnout,and job title is the influencing factor of resilience level.Conclusion Medical workers have high levels of burnout and resilience.Improving working conditions can improve their resilience and reduce burnout.The main influencing factor of burnout is income satisfation,and the main influencing factors of resilience are income satisfaction and professional title.

Clinical application of doxorubicin (DOX) is heavily hindered by DOX cardiotoxicity. Several theories were postulated for DOX cardiotoxicity including DNA damage and DNA damage response (DDR), although the mechanism(s) involved remains to be elucidated. This study evaluated the potential role of TBC domain family member 15 (TBC1D15) in DOX cardiotoxicity. Tamoxifen-induced cardiac-specific Tbc1d15 knockout (Tbc1d15^CKO) or Tbc1d15 knockin (Tbc1d15^CKI) male mice were challenged with a single dose of DOX prior to cardiac assessment 1 week or 4 weeks following DOX challenge. Adenoviruses encoding TBC1D15 or containing shRNA targeting Tbc1d15 were used for Tbc1d15 overexpression or knockdown in isolated primary mouse cardiomyocytes. Our results revealed that DOX evoked upregulation of TBC1D15 with compromised myocardial function and overt mortality, the effects of which were ameliorated and accentuated by Tbc1d15 deletion and Tbc1d15 overexpression, respectively. DOX overtly evoked apoptotic cell death, the effect of which was alleviated and exacerbated by Tbc1d15 knockout and overexpression, respectively. Meanwhile, DOX provoked mitochondrial membrane potential collapse, oxidative stress and DNA damage, the effects of which were mitigated and exacerbated by Tbc1d15 knockdown and overexpression, respectively. Further scrutiny revealed that TBC1D15 fostered cytosolic accumulation of the cardinal DDR element DNA-dependent protein kinase catalytic subunit (DNA-PKcs). Liquid chromatography-tandem mass spectrometry and co-immunoprecipitation denoted an interaction between TBC1D15 and DNA-PKcs at the segment 594-624 of TBC1D15. Moreover, overexpression of TBC1D15 mutant (∆594-624, deletion of segment 594-624) failed to elicit accentuation of DOX-induced cytosolic retention of DNA-PKcs, DNA damage and cardiomyocyte apoptosis by TBC1D15 wild type. However, Tbc1d15 deletion ameliorated DOX-induced cardiomyocyte contractile anomalies, apoptosis, mitochondrial anomalies, DNA damage and cytosolic DNA-PKcs accumulation, which were canceled off by DNA-PKcs inhibition or ATM activation. Taken together, our findings denoted a pivotal role for TBC1D15 in DOX-induced DNA damage, mitochondrial injury, and apoptosis possibly through binding with DNA-PKcs and thus gate-keeping its cytosolic retention, a route to accentuation of cardiac contractile dysfunction in DOX-induced cardiotoxicity.

Objective:To understand the mediating role of resourcefulness between perceived stress and depression in primary caregivers of stroke patients.Methods:The primary caregivers of 311 inpatients with stroke in three Class Ⅲ Grade A hospitals in Yinchuan from December 2020 to August 2021 were selected by convenience sampling and investigated with the general information form, Resourcefulness Scale (RS) , Chinese Perceived Stress Scale (CPSS) and Self-Rating Depression Scale (SDS) . SPSS 25.0 and PROCESS were used to analyze the relationship between the three, and to explore the mediating role of resourcefulness between perceived stress and depression in the primary caregivers of stroke patients.Results:In this study, 311 questionnaires were distributed, 311 questionnaires were recovered, and 308 questionnaires were valid. The primary caregivers of stroke patients scored (24.98±6.38) for perceived stress, (83.98±12.85) for resourcefulness, and (44.35±8.40) for depression. Resourcefulness was negatively correlated with perceived stress and depression scores ( r=-0.313, -0.317; P <0.01) , and perceived stress was positively correlated with depression ( r=0.425, P<0.01) . The Bootstrap method further validated that resourcefulness played a partial mediating role between perceived stress and depression, and the mediating effect accounted for 14.95% of the total effect. Conclusions:Resourcefulness is a mediating variable between the perceived stress and depression of the primary caregivers of stroke patients, and resourcefulness plays a partial mediating role between perceived stress and depression.

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 investigate the application value of Information-Motivation-Behavioral Skill Model (IMB)-based continuing care on the recovery of nerve function and life quality in patients with stroke after thrombolytic therapy. Methods: Stroke patients who underwent thrombolytic therapy were randomly assigned to IMB group(35 cases) and control group(35 cases) according to the method of random number table. The control group recieved routine thrombolysis nursing, IMB group was given IMB-based continuing care. After six months of intervention, nerve function was evaluated by National Institute of Health Stroke Scale(NIHSS) and Modified Rankin Scale(MRS), the active ability was performed by Fugl-Meyer Assessment Scale(FMA) and Barthel Index(BI), the quality of life was assessed by the item short from health survey(SF-36), respectively. Results: Before intervention, the scores of NIHSS, MRS, FMA, BI, SF-36 between two groups was no significant difference (P>0.05). After three and six months of nursing, the NIHSS, MRS was (7.84±2.20), (5.00±1.60) points and (3.48±0.84), (3.07±0.69) points in IMB group, and (9.75±1.82), (8.21±1.37) points and (4.06±1.08), (3.91±0.71) points in control group, there was significant difference between two groups (t =2.417-8.647, P<0.01 or 0.05). After one, three and six months of intervention, FMA was (65.86±5.67), (76.41±8.47), (78.79±8.58) points, BI was (46.29±7.29), (58.09±10.20), (67.50±9.44) points, SF-36 was (33.13±4.64), (43.09±6.70), (49.83±8.56) points in IMB group, (58.53±8.92), (64.47±7.56), (71.81±8.90) points, (42.47±5.67), (48.74±5.39), (56.03±6.55) points and (29.63±4.06), (35.91±5.93), (41.02±9.05) points in control group, there was significant difference between two groups (t =2.333-5.972, P< 0.05). Moreover, in the repeated measures ANOVA showed the differences of NIHSS, MRS, FMA, BI, SF-36 were statistically significant for the group by time interaction(F=13.556-133.994, P<0.05). Conclusions IMB-based continuing care can promotes the recovery of nerve function and improves the quality of life in patients with stroke after thrombolytic therapy.

Objective To observe the clinical features of retinal arterial occlusion (RAO) in youth.Methods This is a retrospective case review.Nine patients (9 eyes) with RAO were enrolled in this study.There were 6 males (6 eyes) and 3 females (3 eyes).The average age was (14.22± 3.93) years.The best-corrected visual acuity (BCVA),indirect ophthalmoscopy,fundus color photography and fundus fluorescein angiography were performed.All patients underwent systemic evaluation including blood routine,erythrocyte sedimentation rate,blood lipids,vasculitis screening,homocysteine level,antiphospholipid antibody,blood coagulation,neck vascular ultrasound,and cardiac color ultrasound and electrocardiogram examination.All patients received oxygen therapy,blood medications and symptomatic treatment.Meanwhile,the patients with autoimmune diseases were received systemic glucocorticoid therapy.The follow-up was ranged from 6 to 12 months.The visual acuity and fundus change before and after treatment were compared.Results among 9 patients,one patient had systemic lupus erythematosus,one patient had congenital heart disease,one patient had hypergammaglobulinemia,and carotid artery color ultrasonography showed that the internal carotid artery vessels faltered in 2 cases.The BCVA was 0.01-0.12.Among 9 eyes,there were 5 eyes (55.6％) with retinal branch artery occlusion (BRAO),2 eyes (22.2％) with central retinal artery occlusion (CRAO),2 eyes (22.2％) with ciliary retinal artery occlusion (CLAO).CRAO eyes showed positive RAPD (relative afferent pupillary defect),fine retinal artery and the corresponding vein,pale white retinal edema in posterior area and macular cherry-red spot.BRAO eyes manifested as inferior temporal artery occlusion and pale white retinal edema around them.CLAO eyes showed temporal ligulate grey-white retinal edema.At the last follow-up,BCVA improved and retinal vessels returned to normal in 7 eyes (77.8％);BCVA unchanged and no improvement in fundus in 2 eyes (22.2％).Conclusion Adolescent RAO is mostly partial occlusion,the prognosis is generally good after early active treatment.

Objective To explore the rehabilitation and outcome of motor neuron syndrome and Sjögren's syndrome. Methods A patient with motor neuron syndrome and Sjögren's syndrome was reviewed. Results The patient expressed progressive quadriparesis and slurred speech. Electromyogramphy displayed upper and lower motor neuron damage. Sonography showed diffuse lesions in parotid gland and submandibular gland. Laboratory tests displayed markedly elevated levels of SSA and Ro52 antibodies. After a course of intravenous immunoglobulin (IVIG) and four weeks of comprehensive rehabilitation, the patient improved in terms of speech, muscle strength, balance, posture control and activities of daily living. She failed to achieve further improvement after another IVIG and glucocorticoid two months later, but fortunately her symptoms did not deteriorate. Conclusion The patient with motor neuron syndrome and Sjögren's syndrome might benefit from IVIG and appropriate rehabilitation.

OBJECTIVETo analyze the outcome of chromosomal microarray analysis (CMA) in prenatal diagnosis for fetal abnormalities detected by ultrasonography.

METHODSAmniotic fluid samples from 477 pregnancies with abnormal ultrasound findings but without common aneuploidies were detected by CMA with Affymetrix CytoScan 750K arrays. The results were analyzed with ChAS v3.0 software.

RESULTSAmong the 477 samples, 24 (5.03%) were detected with pathogenic copy number variations (pCNVs) by CMA. Six (9.68%) among 62 cases with structural fetal abnormalities in multiple organ systems were detected with pCNVs, 11 (7.48%) among 147 cases with a single structural anomaly were detected with pCNVs, and 7 (2.61%) among 268 cases with a soft marker were detected with pCNVs.

CONCLUSIONCMA has offered a clear advantage over conventional karyotyping for the detection of fetal chromosomal abnormalities, and can provide an effective diagnostic tool for those with one or more structural abnormalities detected by ultrasound.

Adolescent ; Chromosome Aberrations ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; DNA Copy Number Variations ; Female ; Fetal Diseases ; diagnosis ; diagnostic imaging ; genetics ; Fetus ; diagnostic imaging ; Humans ; Karyotyping ; Male ; Microarray Analysis ; methods ; Pregnancy ; Prenatal Diagnosis ; Ultrasonography, Prenatal ; methods ; Young Adult