OBJECTIVE To investigate the effect and mechanism of gracillin from Reineckia carnea on autophagy in non- small cell lung cancer A549 cells. METHODS Using A549 cells as subjects， the effects of different concentrations of gracillin （0.25， 0.5， 1， 2， 4 μmol/L） on the proliferation of cells were detected by CCK-8 after being treated for different time （12， 24， 48 h）. Compared with the control group without medication， the effect of gracillin （2 μmol/L） on the formation of autophagosomes in cells was observed by transmission electron microscope after 24 h of exposure. The aggregation of GFP-LC3 on autophagosome membrane was detected by GFP-LC3 plasmid transfection after being treated with gracillin （0.25， 0.5， 1， 2 μmol/L） for 24 h. Quantitative real-time PCR and Western blot assay were used to detect the mRNA and protein expressions of family with sequence similarity 102 member A（FAM102A）， the expressions of autophagy-related proteins [p62， Beclin-1， microtubule-associated protein 1 light chain 3B （LC3B）]， and the expressions of phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway-related proteins in A549 cells after being treated with gracillin （0.25， 0.5， 1 and 2 μmol/L） for 24 h. RESULTS Gracillin significantly inhibited the proliferation of A549 cells in a concentration- and time-dependent manner. The IC50 was 2.55 μmol/L at 24 h. After 24 h of gracillin treatment， autophagosomes with bilayer membrane structure were found in the cell cytoplasm， and GFP-LC3 green fluorescent spots on autophagosome membrane were obvious， representing an increasing trend as drug concentration. Compared with the control group， mRNA and protein expressions of FAM102A （0.5， 1， 2 μmol/L groups）， protein expression of Beclin-1 （1， 2 μmol/L groups） and LC3B-Ⅱ/LC3B-Ⅰ ratio （2 μmol/L group） were significantly increased in different concentrations of gracillin groups， while the protein expression of p62 （1， 2 μmol/L groups）， and the protein phosphorylations of Akt （1， 2 μmol/L groups） and PI3K （2 μmol/L group） were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS Gracillin can promote excessive autophagy in A549 cells by up-regulating mRNA and protein expressions of FAM102A and inhibiting PI3K/Akt signaling pathway， thus inhibiting cell proliferation.

Objective To analyze the real experiences of patients with inflammatory bowel disease(IBD)during dietary restrictions,providing references for healthcare personnel to guide patients in standardizing dietary restriction behaviors.Methods Purposeful sampling was employed to select 14 patients with IBD who were treated at a tertiary A hospital in Shanghai between October 2022 and February 2023 for semi-structured in-depth interviews.Data were analyzed using the Colaizzi's 7-step method in phenomenological research.Results 4 themes and 13 sub-themes were extracted.Theme 1:facing multiple physiological challenges(hunger,nutritional imbalance,fatigue and muscle atrophy).Theme 2:experiencing negative psychological disturbances(craving and struggle for gourmet food,diminished zest for life and increased anxiety,feeling embarrassed during social activities).Theme 3:adopting various coping strategies(self-adjustment and adaptation to dietary restrictions,satisfying oral desires through various avenues,seeking knowledge and guidance on nutrition).Theme 4:gaining more growth and support(improved symptom and quality of life,enhanced awareness of dietary health,improved ability to manage diet,receiving support from peers and family).Conclusion The experience of dietary restrictions in IBD patients is complex and varied.Nursing staff should prioritize nutritional risk screening for IBD patients,pay attention to their mental health,provide patients with scientific and personalized dietary guidance,and strengthen social and family support to assist patients in better self-management of their diet.

Objective To investigate the interventional effect and mechanism of a novel ampelopsis hydrogel on dampness-heat accumulation syndrome of gouty arthritis. Methods A total of 90 patients with gouty arthritis who met the diagnostic criteria of western medicine and were differentiated as damp-heat accumulation syndrome of traditional Chinese medicine(TCM) were randomly divided into treatment group, control group and blank group, with 30 patients in each group. The blank group was treated with etoricoxib only, the control group was treated with etoricoxib combined with ampelopsis hydrogel, and the treatment group was treated with etoricoxib combined with external application of ampelopsis hydrogel. The clinical efficacy, time to symptom improvement, safety, comfort, changes in syndrome scores of TCM, serum inflammatory factors[C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), erythrocyte sedimentation rate (ESR)], NF-κB signaling pathway-related proteins, Visual Analogue Scale (VAS) scores for pain, and joint mobility were compared among the three groups before and after treatment. Results The total effective rates in the treatment group and control group were 93.33% and 90.00%, respectively, which were higher than 70.00% in the blank group (P < 0.05). The time for improvement of pain, redness, tenderness, and limited joint mobility in the treatment group was shorter than that in the control group and blank group (P < 0.05). After 7 days of treatment, the TCM syndrome score in the treatment group was lower than that in the control group and blank group, and the levels of serum CRP, TNF-α, and ESR and the expressions of NF-κB signalingpathway-related proteins P50 and P65 in the treatment group and control group were lower than those in the blank group (P < 0.05). After 7 days of treatment, the VAS score in the treatment group was lower than that in the control group and blank group, and the comfort score in the treatment group was higher than that in the control group (P < 0.05). There was no significant difference in the incidence of adverse reactions among the three groups (P>0.05). Conclusion The effect of ampelopsis hydrogel in treating gouty arthritis is better than that of ampelopsis paste, and its mechanism may be related to the regulation of the NF-κB pathway and inhibition of inflammatory factor expression. The hydrogel is easy to use, hygienic, and comfortable, and is expected to become a safe, effective, and convenient external medicine for gouty arthritis.

Objective:To investigate the effectiveness of case-based learning (CBL) combined with online teaching in standardized residency training of rheumatology and immunology.Methods:A total of 78 individuals who participated in standardized residency training in Department of Rheumatology and Immunology in our hospital from June 2019 to August 2020 were included and divided into observation group and control group. The individuals in the control group received traditional teaching, and those in the observation group received CBL combined with online teaching. The physicians receiving standardized residency training were evaluated by theoretical examination, clinical operation skill assessment, and instructor rating, and the degree of satisfaction with teaching, degree of satisfaction with teaching methods, and classroom learning atmosphere were also evaluated.Results:The observation group had a theoretical examination score of (94.10±2.01) and a clinical operation skill assessment score of (90.44±1.57), which were significantly higher than those of the control group ( P<0.05), and the observation group had a significantly better instructor rating (89.36±1.33) than the control group ( P<0.05). Compared with the control group, the observation group had significantly higher degree of satisfaction with teaching (3.79±0.41), degree of satisfaction with teaching methods (3.92±0.27), and evaluation of classroom learning atmosphere (3.90±0.31) ( P<0.05). Conclusion:CBL combined with online teaching can help to improve learning efficiency, stimulate the enthusiasm for learning, expand clinical thinking, promote the growth of teaching and learning, and form a virtuous cycle among trainees receiving standardized residency training, which holds promise for further exploration.

Objective:To observe the changes of relevant clinical indicators and ultrasound pre-sentations in rheumatoid arthritis (RA) patients after being treated with tocilizumab for 3 months and explore the efficacy and safety of tocilizumab in the treatment of RA.Methods:Clinical data, laboratory and ultrasound examinations and medications of RA patients who met inclusion criteria in our hospital from Jan-uary 2017 to September 2020 were included and their data were analyzed retrospectively, and the efficacy and safety of tocilizumab and the ultrasound assessment of disease remission were analyzed. Paired sample t test, Wilcoxon signed-rank test, χ2 test or Fisher's exact probability test were used for comparative analysis. Results:① Compared with baseline data, morning stiffness duration of patients treated with tocilizumab for 3 months [60(30, 120) min vs 0(0, 10) min, Z=-6.19, P<0.001], disease activity score of 28 joints-erythrocyte sedimentation rate (DAS28-ESR) [(4.6±1.5) vs (3.2±1.2), t=6.83, P<0.001], disease activity score of 28 joints-C-reactive protein (DAS28-CRP) [(4.2±1.4) vs (2.8±1.1), t=7.14, P<0.001], swollen joint count (SJC) [2(1, 7) vs 0(0, 2), Z=-4.31, P<0.001], tender joints count (TJC) [6(2, 13.5) vs 2(0,4), Z=-5.16, P<0.001], gray scale score (GS) [4.5(2, 6) vs 1(0, 3), Z=-5.86, P<0.001], intra-synovial blood flow energy Doppler (PD) [2(1, 3) vs 0(0, 0), Z=-5.38, P<0.001], white blood cell (WBC) [6.6(4.9, 8.4)×10 9/L vs 5.7(4.9, 7.3)×10 9/L, Z=-2.83, P=0.005], hemoglobin (Hb) [119(104, 131) g/L vs 123(113, 136) g/L, Z=-2.82, P=0.007], ESR [32(14.5, 50) mm/1 h vs 19 (10, 30) mm/1 h, Z=-3.31, P=0.001], CRP [11.40(3.02, 25.80) mg/L vs 3.49(1.30, 11.82) mg/L, Z=-2.78, P=0.004], D-dimer (D-D) [0.93(0.47, 2.07) mg/L vs 0.43(0.21, 0.80) mg/L, Z=-3.77, P<0.001] were significantly improved, and the difference was statistically significant. ② The serum levels of interleukin (IL)-2 [2.08(1.43, 2.76) pg/ml vs 1.21(0.54, 2.08) pg/ml, Z=-2.67, P=0.008], IL-6 [22.40(5.13, 67.27) pg/ml vs 14.63(5.27, 27.71) pg/ml, Z=-2.81, P=0.005], IL-10 [(2.53±0.68) pg/ml vs (1.74±0.74) pg/ml, t=2.60, P=0.017] were significantly changed, while serum levels of IL-4 [1.63(1.08, 3.38) pg/ml vs 1.33(0.97, 2.59) pg/ml, Z=-0.89, P=0.374], tumor necrosis factor (TNF) -α [4.04(1.41, 10.45) pg/ml vs 1.62(0.84, 3.79) pg/ml, Z=-1.92, P=0.056], IL-17 [4.68(1.67, 6.72) pg/ml vs 3.15(1.81, 5.29) pg/ml, Z=-0.53, P=0.594] were not significantly changed from baseline data. ③ There was poor consistency between ultrasonic response and DAS28-ESR response, simplified disease activity Index (SDAI) response and clinical disease activity index (CDAI) response ( Kappa coefficient: 0.142, 0.142, 0.191), but no consistency between ultrasonic response and DAS28-CRP response (Kappa coefficient: -0.015) were found. Receiver operating characteristic (ROC) curve showed that ultrasound was not statistically significantly different in assessing the remission of RA, indicating subclinical synovitis remained in ultrasound examination even though clinically remission could be reached based on the above scoring indexes in RA patients. ④ In terms of adverse reactions, neutrophils (NEU) of patients after 3 months' tocilizumab treatment [4.47(2.77, 5.39)×10 9/L vs 3.76(2.98, 4.74)×10 9/L, Z=-2.77, P=0.006], platelet count (PLT) [(291±84)×10 9/L vs (254±70)×10 9/L, t=4.76, P<0.001] were significantly decreased, high-density lipoprotein-cholesterol (HDL-C) [(1.22±0.27) mmol/L vs (1.39±0.34) mmol/L, t=3.12, P=0.003], low density lipoprotein-cholesterol (LDL-C) [(1.96±0.66) mmol/L vs (2.19±0.84) mmol/L, t=3.15, P=0.003], triglyceride (TG) [0.85(0.68, 1.08) mmol/L vs 0.93(0.71, 1.25) mmol/L, Z=-2.36, P=0.018] and total cholesterol (TC) [(4.18±1.04) mmol/L vs (4.52±1.16) mmol/L, t=3.33, P=0.002] were significantly different from baseline. Among 65 patients, 5 patients (7.7%) had transaminase abnormality, but returned to normal after symptomatic treatment. Conclusion:Tocilizumab treatment can effectively suppress the inflammatory reactions, improve the clinical symptoms and prognosis of patients, however, patients who judged as clinical remission according to the current clinical commonly scores may still have subclinical active disease, ultrasound results should be included as one criteria for disease remission assessment and take into consideration when adjusting treatrnent.

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.

Objective:To explore the application effect of quantitative evaluation nursing in Operating Room in patients undergoing radical resection of rectal cancer.Methods:A total of 114 patients who received radical resection of rectal cancer in Xi'an Daxing Hospital from December 2019 to December 2021 were selected by the convenient sampling method. According to the order of admission, they were divided into the observation group and the conventional group, with 57 cases in each group. The observation group received quantitative evaluation nursing in the Operating Room, while the conventional group received routine Operating Room nursing. The differences in physiological indexes and mood state scores were compared between the two groups.Results:The postoperative diastolic blood pressure, systolic blood pressure and heart rate in the observation group were lower than those in the conventional group ( P<0.05). The scores of depression-frustration, tension-anxiety, fatigue-dullness, anger-hostility and confusion-chaos dimensions of postoperative mood state in the observation group were lower than those in the conventional group, and the energy-vitality score was higher than that in the conventional group ( P<0.05) . Conclusions:Quantitative evaluation nursing in the Operating Room can reduce the psychosomatic stress response in patients undergoing radical resection of rectal cancer.

Objective: To explore the clinical value of serum autoantibodies and human leukocyte antigen (HLA-B₂₇) molecular testing in Uygur patients with human immunodeficiency virus (HIV) infection. Method: A total of 727 HIV-infected Uygur patients who visited Kuche infectious diseases hospital during May 2016 to March 2017 were include in this study. The other 390 healthy people were enrolled as controls. Serum antinuclear antibodies (ANA), anti-cyclic citrullinated peptide (CCP) antibody, anti-extractable nuclear antigen (ENAs) antibody and HLA-B₂₇ molecule were tested. Result: Among 727 HIV-infected Uygur patients, 317 were males and 410 were females with mean age (35.52±13.44) years old. The mean duration of disease was (6.34±3.05) years. There were 697 (95.87%) patients receiving highly active antiretroviral therapy (HAART) with mean duration of treatment (5.52±3.47) years. The mean CD4⁺T cell count was (520±271) cells/μl in 202 HIV-infected patients, and mean virus load was (108 139±20 498) copies/ml in 20 HIV-infected patients. Rheumatic manifestations were recorded in 238 (32.74%) HIV-infected Uygur patients, mainly with dry mouth and dry eye (15.41%) , alopecia (9.90%) , arthralgia (8.94%) , ect. Compared with the health controls, positive ANA was more common in HIV infected Uygur patients (33.43% vs. 17.43%, P<0.001) with low titers (ANA titer:1∶100) . HIV-infected Uygur patients had higher positive anti-u1-RNP antibodies positive rate (1.10%), but lower anti-SSA antibodies positive rate (0.14%) and anti-CCP antibodies positive rate (0.28%). Patients with positive ANA in HAART group were significantly less than that in non-treatment group (32.71% vs. 50.00%, P=0.049). There were no correlations between ANA and duration of HAART, CD4⁺T cell counts and virus load (r values 0.061, 0.047, 0.121, respectively. P>0.05). Only one female patient was HLA-B₂₇ positive (0.14%), which was significantly lower than that in healthy controls (3.08%) (P<0.001). Also, only one patient was diagnosed with rheumatoid arthritis (RA). Conclusion Autoimmune manifestations are common in HIV-infected Uygur patients. Several autoantibodies are positive, but the coincidence of rheumatic diseases is rare. It′s noted that patients with autoimmune manifestations should be considered as a differential diagnosis of HIV infection.

As a newly-identified protein post-translational modification, malonylation is involved in a variety of biological functions. Recognizing malonylation sites in substrates represents an initial but crucial step in elucidating the molecular mechanisms underlying protein malonylation. In this study, we constructed a deep learning (DL) network classifier based on long short-term memory (LSTM) with word embedding (LSTM) for the prediction of mammalian malonylation sites. LSTM performs better than traditional classifiers developed with common pre-defined feature encodings or a DL classifier based on LSTM with a one-hot vector. The performance of LSTM is sensitive to the size of the training set, but this limitation can be overcome by integration with a traditional machine learning (ML) classifier. Accordingly, an integrated approach called LEMP was developed, which includes LSTM and the random forest classifier with a novel encoding of enhanced amino acid content. LEMP performs not only better than the individual classifiers but also superior to the currently-available malonylation predictors. Additionally, it demonstrates a promising performance with a low false positive rate, which is highly useful in the prediction application. Overall, LEMP is a useful tool for easily identifying malonylation sites with high confidence. LEMP is available at http://www.bioinfogo.org/lemp.
Amino Acid Sequence ; genetics ; Amino Acids ; Animals ; Deep Learning ; Forecasting ; methods ; Lysine ; chemistry ; Machine Learning ; Malonates ; chemistry ; Protein Processing, Post-Translational ; genetics

Esophageal squamous-cell carcinoma (ESCC) is one of the most lethal malignancies in the world and occurs at particularly higher frequency in China. While several genome-wide association studies (GWAS) of germline variants and whole-genome or whole-exome sequencing studies of somatic mutations in ESCC have been published, there is no comprehensive database publically available for this cancer. Here, we developed the Chinese Cancer Genomic Database-Esophageal Squamous Cell Carcinoma (CCGD-ESCC) database, which contains the associations of 69,593 single nucleotide polymorphisms (SNPs) with ESCC risk in 2022 cases and 2039 controls, survival time of 1006 ESCC patients (survival GWAS) and gene expression (expression quantitative trait loci, eQTL) in 94 ESCC patients. Moreover, this database also provides the associations between 8833 somatic mutations and survival time in 675 ESCC patients. Our user-friendly database is a resource useful for biologists and oncologists not only in identifying the associations of genetic variants or somatic mutations with the development and progression of ESCC but also in studying the underlying mechanisms for tumorigenesis of the cancer. CCGD-ESCC is freely accessible at http://db.cbi.pku.edu.cn/ccgd/ESCCdb.
Aged ; Asian Continental Ancestry Group ; genetics ; China ; epidemiology ; Databases, Genetic ; Esophageal Squamous Cell Carcinoma ; genetics ; Female ; Genetic Predisposition to Disease ; Genetic Variation ; Genome-Wide Association Study ; Humans ; Internet ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; User-Computer Interface