[摘 要] 目的：探索淋巴细胞亚群对非小细胞肺癌（NSCLC）患者接受程序性死亡受体1（PD-1）单抗联合化疗的疗效预测及预后评估的价值。方法：回顾性分析2022年1月至2023年12月在兰州大学第二医院确诊的接受PD-1单抗联合化疗的50例NSCLC患者的临床资料，收集患者治疗前及治疗2周期后的外周血淋巴细胞亚群（包括总T细胞、CD4+ T细胞、CD8+ T细胞、NK细胞、总B淋巴细胞、CD4+/CD8+ T细胞比值等）的数据。治疗2周期后进行影像学检查评价治疗的疗效，分为疾病控制（DC）组和疾病进展（PD）组。使用卡方检验、秩和检验和Logistic回归分析淋巴细胞亚群表达水平与NSCLC患者近期疗效的关系，采用Kaplan-Meier法分析无进展生存期（PFS）预测疗效的价值。结果：PD-1单抗联合化疗对NSCLC患者的免疫状态产生了显著影响，接受免疫联合化疗后，患者外周血CD4+ T细胞、CD4+/CD8+ T细胞比值均显著升高（均P < 0.01），CD8+ T细胞下降。近期疗效显示，DC组患者血清CD4+ T细胞比例及CD4+/CD8+ T细胞比值均高于PD组（均P < 0.01）。Logistic多因素分析显示，CD4+/CD8+ T细胞比值是PD-1单抗联合化疗疗效的独立影响因素。通过ROC曲线分析，CD4+/CD8+ T细胞比值变化量AUC为0.820 > 0.5，截断值为0.15，CD4+/CD8+ T细胞比值变化量 ≥ 0.15的患者的PFS更长。结论：晚期NSCLC患者外周血中CD4+ T细胞和CD8+ T细胞比例、CD4+/CD8+ T细胞比值可以预测PD-1单抗联合化疗的疗效和预后。

Biochemical liver function tests are important methods to determine liver function in clinical practice， but abnormal liver biochemical parameters are not completely equivalent to liver damage. Some genetic and immune factors can also cause abnormal liver biochemical parameters， but with good prognosis in most cases. This article summarizes the causes of some benign abnormal liver biochemical parameters， so as to help clinicians to broaden their thinking of diagnosis and treatment， take into account genetic and immune factors， and avoid misdiagnosis and mistreatment.

Morita therapy has been bom for more than 100 years.Inpatient Morita therapy is highly oper-able and easy to master.It can improve many refractory neuroses through four-stage treatment.But more neuroses are treated in outpatient clinics,and Morita therapy cannot be used in hospitalized patients.Therefore,the formula-tion of expert opinions on outpatient operations is particularly important.This paper is based on domestic and for-eign references,and after many discussions by domestic Morita therapy experts,and then drew up the first version of the expert opinions on operation of outpatient Morita therapy.Meanwhile the operation rule of Morita therapy in three stages of outpatient treatment was formulated:in the etiological analysis stage,under the theoretical guidance of Morita therapy,analyze the pathogenic factors,to improve treatment compliance and reduce resistance;during the operating stage,guide patients to engage in constructive and meaningful actions,realizing the achievement of letting nature take its course principle;in the cultivating character and enriching life stage,pay attention to positive infor-mation,expanding the scope and content of actions,improving the ability to adapt to complex life,and preventing recurrence caused by insufficient abilities.It will lay a foundation for the promotion of Morita therapy in domestic outpatient clinics,so that more patients with neurosis and other psychological diseases could receive characteristic Morita therapy treatment in outpatient clinics.

Objective To investigate the effect of agomelatine on the clinical efficacy in elderly patients with acute cerebral infarction(ACI)and comorbid anxiety-depression disorders by regu-lating serum neurotransmitters and nerve cytokines.Methods A total of 160 elderly ACI patients with anxiety and depression symptoms admitted in Pingxiang Second People's Hospital from June 2020 to December 2023 were enrolled in this study.All of them received thrombolysis or interven-tional therapy,and then were randomly divided into control and observation groups,with 80 patients in each group.The control group received conventional psychological intervention,while the observation group was given additional oral administration of agomelatine.Cognitive function,neurological function,neurotransmitters and neuronal cytokines,anxiety and depression scores,sleep quality,quality of life,daily activity ability and adverse reactions were compared between the two groups.Results After intervention,Mini Mental State Examination(MMSE)scores,levels of neuropeptide Y(NPY),5-hydroxytryptamine(5-HT),norepinephrine(NE),dopamine and brain-derived neurotrophic factor(BDNF),36-item Brief Health Questionnaire(SF-36)score,and Bar-thel index scale score were significantly higher in both 2 groups when compared with above indicators before the intervention(P＜0.01).And the MMSE score,NPY,5-HT,NE,dopamine and BDNF levels,SF-36 score and Barthel index scale score were obviously higher in the observa-tion group than the control group(P＜0.01).Both groups obtained notably lower NIHSS score,S100 calcium binding protein B(S100B)and myelin basic protein(MBP)levels,Hamilton Anxiety Rating Scale(HAMA)score,and Hamilton Depression Rating Scale 17(H AMD-17)score and Pittsburgh Sleep Quality Index(PSQI)score after intervention(P＜0.01).And the NIHSS score,S100B and MBP levels,HAMA score,HAMD-17 score,and PSQI score were statistically lower in the observation group than the control group(P＜0.01).During the treatment process,no signifi-cant difference was observed in the incidence of total adverse reactions between the two groups(3.75％vs 6.25％,x2=0.526,P=0.468).Conclusion When agomelatine tablets are indicated for ACI patients with concomitant anxiety-depression disorders,they can effectively rehabilitate cog-nitive function,enhance neurological function,improve sleep quality and quality of life,optimize activities of daily living,eliminate negative emotions,and correct the expression of neurotransmit-ters and neurotrophic factors.

Objective:To develop the anti-CD30 monoclonal antibody 64Cu-1, 4, 7-trizacyclononane-1, 4, 7-triacetic acid (NOTA)-CD30 and visualize CD30 expression in lymphoma non-invasively. Methods:The CD30 expression levels of 5 cell lines (Karpas299, Raji, Daudi, Ramos, and U266) were assessed by Western blot. Cell lines with high and low CD30 expression were selected for flow cytometry to evaluate the specific binding affinity of anti-CD30 monoclonal antibody. Thirteen NSG mice were used to established CD30 positive and negative subcutaneous xenograft models. 64Cu-NOTA-CD30 was obtained and 64Cu-NOTA-immunoglobulin (Ig)G was used as the control. ImmunoPET imaging was performed 2, 24, and 48 h after the injection of 64Cu-NOTA-CD30 or 64Cu-NOTA-IgG. Finally, the biodistribution studies were conducted. Repeated-measures analysis of variance and Bonferroni test were conducted for comparison. Results:Karpas299 showed the highest CD30 expression, while Raji showed the lowest. Flow cytometry showed specific binding affinity of the anti-CD30 monoclonal antibody to the Karpas299 cell line. The radiochemical purities of the probes were both higher than 95%. In microPET, the 64Cu-NOTA-CD30 uptake of Karpas299 xenograft tumors increased over time, with (11.46±0.58), (17.60±1.16) and (19.46±0.99) percentage activity of injection dose per gram of tissue (%ID/g) at 2, 24 and 48 h respectively. The contrast to normal tissue was good at 48 h, with the tumor/heart (blood) ratio of 2.20±0.22. The uptake of 64Cu-NOTA-CD30 in Karpas299 tumor at 48 h after injection was significantly higher than that in Raji tumor ((6.10±1.03) %ID/g) and 64Cu-NOTA-IgG in Karpas299 tumor ((5.12±0.89) %ID/g; F=290.99, t values: 19.65 and 22.25, all P<0.001). The uptake of 64Cu-NOTA-CD30 and the control probe in the heart and liver decreased over time in all groups. Ex vivo biodistribution at 48 h was mainly consistent with the results of microPET in vivo. Conclusions:64Cu-NOTA-CD30 is able to visualize the expression level and distribution of CD30 non-invasively. It is promising to be applied for screening the beneficial groups and evaluating efficacy for CD30-targeted immunotherapy.

Objective:To investigate the effects of circ_000543 derived from hypoxic exosomes on proliferation and invasion of breast cancer (BC) cells.Methods:Bioinformatic website was used to predict the abnormal expression of circ_000543 in BC tissue and the transcription factor that might regulate circ_000543. Double luciferase report experiment and ChIP assay were used to confirm the regulation relationship between YY1 and circ_000543. Exosomes were separated from normal BC cells and BC cells under hypoxic condition, and qRT-PCR was adopted to detect the expression of circ_000543 in exosomes. The expression of circ_000543 and YY1 in exosomes was intervened and the exosomes were cocultured with BC cells under normoxia. CCK8 and Transwell assay was used to detect the proliferation and invasive ability individually.Results:qRT-PCR experiment found that, compared with MCF-10A cells (1±0.11) and exosomes isolated from normoxic cells (1±0.10), circ_000543 expression was up-regulated in BC cells (1.59±0.13) and exosomes derived from cells under hypoxic condition (1.63±0.12) ( t=6.001, P=0.004; t=6.986, P=0.002). Exosomes derived from cells under hypoxic condition promoted proliferation and invasion of BC cells under normoxia. Inhibition of circ_000543 partially offset the effects of exosomes. YY1 induced the expression of circ_000543 in BC cells as a transcription factor. The expression of circ_000543 was inhibited when YY1 expression was down-regulated; at the same time, down-regulation of YY1 inhibited the effects of exosomes on proliferation and invasion of BC cells. Conclusion:The transcription factor YY1 promoted proliferation and invasion of BC cells by inducing hypoxic breast cancer-derived exosomal circ_000543.

Objective:To investigate the effect of methyltransferase like 14 (METTL14) on the proliferation and invasion of breast cancer (BC) cells by regulating cyclin L2 (Cyclin L2, CCNL2) through m6A modification.Methods:Cancer tissues and paracancerous tissues of BC patients in Yantaishan hospital were collected from Aug. 2018 to Feb. 2020. The expression levels of m6A, METTL14 and CCNL2 in tissues were detected by high performance liquid chromatography/mass spectrometry (HPLC/MS) and qRT-PCR. Dual-luciferase reporter assay, qRT-PCR, and western blot were used to verify the regulatory relationship between METTL14 and CCNL2. RIP experiments verified the regulatory relationship between YTH domain-containing family protein (YTHDF2) and CCNL2. Cell viability was detected by MTT method, and cell invasion ability was detected by Transwell.Results:Compared with normal cells (0.24±0.02) and tissues (0.18±0.02) , BC cells MCF-10A (0.47±0.03, t=11.05, P<0.001) and HS-578T (0.41±0.03, t=8.17, P=0.001) and BC tissues (0.39±0.02, t=12.86, P<0.001) m6A level increased. Compared with normal tissues (1.00±0.26) (0.84±0.07) , METTL14 mRNA (1.57±0.28, t=13.50, P<0.001) and protein levels (1.66±0.11, t=10.89, P<0.001) in BC tissues were significantly increased high. Compared with the control group (100.00±10.11) (1.00±0.12) , the BC cell invasion ability (54.15±6.21, t=6.69, P=0.003) and activity (0.64±0.06, t=4.65, P=0.010) were weakened. Compared with the control group (100±11.05) (1±0.13) , the BC cell invasion ability (175.31±13.45, t=7.49, P=0.002) and activity (2.16±0.16, t=9.75, P=0.002) in the METTL14 overexpression group were enhanced, and the effects of METTL14 on cell invasion (137.41±12.64, t=3.56, P=0.024) and activity (1.64±0.15, t=5.59, P=0.005) were partially reversed after m6A inhibitor treatment change. Compared with normal tissues, CCNL2 expression was down-regulated in BC tissues, and the interaction between CCNL2 and METTL14 was confirmed. Compared with the control group (1.00±0.1) (0.64±0.05) , knockdown of METTL14 could make CCNL2 mRNA (1.67±0.05) . 0.13, t=7.08, P=0.002) and protein (1.09±0.09, t=7.57, P=0.002) were up-regulated. METTL14 knockout enhanced the stability of CCNL2 mRNA through a YTHDF2-dependent pathway, compared with sh-METTL14 group (50.47±5.16) (0.52±0.05) , BC cell invasion ability of sh-METTL14+sh-CCNL2 group (71.69±6.41, t=4.47, P=0.011) and activity (0.64±0.05, t=2.94, P=0.042) were improved. Conclusion:METTL14 inhibits the expression of CCNL2 through m6A modification to enhance the invasion and activity of BC cells.

Objective:To understand the classification characteristics of psychological resilience of nursing undergraduates and analyze the influencing factors among different categories.Methods:Using the convenient sampling method, a total of 506 four-year undergraduate nursing students from the Daqing campus of Harbin Medical University were selected as the research objects in October 2019. General information questionnaire and Connor-Davidson Resilience Scale (CD-RISC) were used to investigate them.Results:The characteristics of psychological resilience of nursing undergraduates could be divided into low resilience group (14.5%) , general resilience group (40.9%) , social avoidance group (22.5%) and high resilience group (22.1%) . Binomial logistic regression analysis results showed that gender, family average monthly income, source place of students and applying satisfaction were the influencing factors of the potential categories of psychological resilience.Conclusions:The psychological resilience of nursing undergraduates has obvious classification characteristics. There are different distributions of nursing undergraduates in different potential categories by gender, family average monthly income, source place of students and applying satisfaction. Teachers should formulate hierarchical interventions according to relevant influencing factors to improve the psychological resilience of nursing undergraduates.

Objective To evaluate the effect of photoaging on the degradation of advanced glycation end products (AGEs) by human dermal fibroblasts.Methods Some cultured human dermal fibroblasts were subjected to repetitive ultraviolet A (UVA) radiation (UVA radiation group) to establish a photoaging cell model,which was then evaluated by cell counting kit 8 (CCK-8) assay,senescenceassociated β-galactosidase staining and detection of apoptosis rate.Moreover,fibroblasts receiving no treatment served as control group.Some other primary fibroblasts were divided into 4 groups:photoaged group receiving UVA radiation,non-photoaged group receiving no treatment,AGE-treated photoaged group treated with UVA radiation followed by the treatment with 200 mg/L AGE-bovine serum albumin (BSA),and AGE-treated non-photoaged group treated with 200 mg/L AGE-BSA alone.After the treatment with AGE-BSA for 4-72 hours,flow cytometry was performed to determine the fluorescence intensity of AGE-BSA in fibroblasts of the above groups.After 8-hour treatment with AGE-BSA,confocal laser scanning microscopy was performed to localize and semiquantitatively detect AGE-BSA in fibroblasts,and enzymelinked immunosorbent assay (ELISA) was conducted to detect AGE-BSA levels in fibroblasts,as well as changes in the intracellular AGE-BSA level within 24 hours after the removal of AGE-BSA.Results Compared with the control group,the UVA radiation group showed significantly decreased cellular proliferative activity (t =7.559,P ＜ 0.05),but significantly increased apoptosis rate and percentage of β-galactosidase-positive fibroblasts (t =14.075,43.524 respectively,both P ＜ 0.05).Flow cytometry revealed that the average fluorescence intensities of AGE-BSA after 4-,8-,16-,24-,48-and 72-hour treatment with AGE-BSA were significantly higher in the AGE-treated photoaged group (293.00 ± 8.19,359.67 ± 11.59,347.00 ± 12.29,338.00 ± 12.77,334.67 ± 14.22 and 336.30 ± 10.21,respectively) than in the photoaged group (all P ＜ 0.05),as well as in the AGE-treated non-photoaged group (222.33 ± 8.74,276.33 ± 6.11,256.33 ± 5.51,243.00 ± 10.15,236.33 ± 1.53 and 240.33 ± 1.52,respectively) than in the non-photoaged group (all P ＜ 0.05).Moreover,the average fluorescence intensities of AGE-BSA at different time points were all significantly higher in the AGE-treated photoaged group than in the AGE-treated non-photoaged group (all P ＜ 0.05).Confocal laser scanning microscopy showed that AGE-BSA was mainly localized in lysosomes after endocytic uptake into the fibroblasts,and the AGE-treated photoaged group showed significantly increased fluorescence intensity of AGE-BSA compared with the AGE-treated non-photoaged group (P ＜ 0.05).ELISA revealed that the intracellular AGE level in the AGE-treated non-photoaged group at 24 hours after the removal of AGE-BSA was decreased by (14.6 ± 1.2)％ compared with that before the removal,and the degradation rate of AGE-BSA was significantly higher in the AGE-treated non-photoaged group than in the AGE-treated photoaged group (7.6％ ± 1.4％,t =6.604,P ＜ 0.05).Conclusion The internalized AGE-degradating ability decreases in photoaged fibroblasts,which may induce the accumulation of AGEs in photoaged skin.

Objective To determine the expression of cathepsin D and advanced glycation end products (AGEs)in skin tissues from patients of different ages or skin tissues with different degrees of sun exposure,to evaluate their correlation,and to preliminarily investigate the role of cathepsin D in the degradation and accumulation of AGEs in photoaged skin.Methods Skin tissues were collected from sunexposed and sun-protected body sites in patients aged 15-20 years,35-40 years,55-60 years or 75-80 years.These skin tissues were divided into 8 groups according to age of patients and degrees of sun exposure,and there were 6 specimens in each group.Immunohistochemical and immunofluorescent methods were used to measure the expression of cathepsin D and AGEs in the skin tissues.Statistical analysis was carried out by factorial design analysis of variance,Wilcoxon rank sum test and Kruskal-Wallis rank sum test for analyzing associations of the expression of cathepsin D and AGEs with age and sun exposure,as well as by Pearson correlation analysis for assessing the correlation between cathepsin D expression and AGEs expression.Results Immunohistochemical study showed that the expression of cathepsin D markedly decreased along with the increase of age,but the accumulation of AGEs gradually increased along with the increase of age.In the same age group,the cathepsin D expression was lower in the sun-exposed skin tissues than in the sun-protected skin tissues,while the accumulation of AGEs was more in the sun-exposed skin tissues than in the sun-protected skin tissues.Factorial design analysis of variance showed that sun exposure could decrease the expression of cathepsin D (F =58.70,P ＜ 0.001),but increase the accumulation of AGEs (F =158.18,P ＜ 0.001).Moreover,the increase of age could lead to decreased expression of cathepsin D (F =79.49,P ＜ 0.001),and increased expression of AGEs (F =106.06,P ＜0.001).Compared with the sun-protected skin tissues,the sun-exposed skin tissues in all the age groups showed significantly lower absorbance value of cathepsin D (35-40 years:0.020 ± 0.005 vs.0.032 ± 0.005;55-60 years:0.012 ± 0.004 vs.0.026 ± 0.002;75-80 years:0.002 ± 0.001 vs.0.013 ± 0.004;all P ＜0.001),but higher absorbance value of AGEs (35-40 years:0.030 ± 0.008 vs.0.010 ± 0.003;55-60years:0.066 ± 0.010 vs.0.021 ± 0.004;75-80 years:0.085 ± 0.015 vs.0.035 ± 0.009;all P ＜ 0.001)except the age group of 15-20 years.No matter whether the skin tissues were sun-exposed or sunprotected,there were significant differences in the expression of cathepsin D and AGEs among different age groups (all P ＜ 0.001).The results of double immunofluorescence staining were similar to those of immunohistochemical study.Pearson correlation analysis showed that the expression of cathepsin D in the sun-exposed skin tissues was highly negatively correlated with the accumulation of AGEs (r =-0.915,P ＜0.05),while they were moderately negatively correlated in the sun-protected skin tissues (r =-0.730,P ＜0.05).Conclusions Along with the increase of age,the expression of cathepsin D in skin tissues decreased,but the expression of AGEs increased.In the sun-protected skin tissues,the expression of cathepsin D was moderately negatively correlated with the expression of AGEs,while they were highly negatively correlated in the sun-exposed skin tissues,suggesting that cathepsin D may play an important role in the degradation and accumulation of AGEs in photoaged skin.