Lysosomes represent a promising target for cancer therapy and reducing drug resistance. However, the short treatment time and low efficiency of lysosomal targeting have limited the application in lysosome-targeting anticancer drugs. In this study, we proposed an adhesive-bandage approach and synthesized a new lysosomal targeting drug, namely long-term lysosome-targeting anticancer drug (LLAD). It contains a SLC38A9-targeting covalently bound moiety and an alkaline component both to prolong the inhibition of SLC38A9 in lysosomes and alkalinize lysosomes. Upon short term and low-dose treatment of HeLa cells, at passage 0, with LLAD, it rapidly alkalinized lysosomes and also can be detected in lysosomes even at passage 15. LLAD induced apoptosis in HeLa cells through long-term lysosomal damage, and showed better long-term anticancer effect than cisplatin in vivo. Overall, our study paves the way for developing long-term lysosomal targeting drugs to treat cancer and overcome the drug resistance of cancer cells, and also provides a candidate drug, LLAD, for treating cancer.

Objective:To investigate the application value of intelligent laboratory construction in the laboratory.Methods:All samples sent to the Department of Clinical Laboratory from all the clinical departments at the Eighth Affiliated Hospital of Sun Yat-Sen University from January 2018 to June 2024 were collected. Full-process intelligentization building was achieved by the use of logistics transmission system, automatic sampling system, automatic quality control system, automatic audit system, intelligent monitoring system, critical value monitoring system, and intelligent DxlabReport operation management system, et al. The normal distribution data were analyzed by two independent samples t-test, and the skew distribution data were analyzed by Mann-Whitney U test. Results:After the implementation of the pneumatic logistics transmission system, the turn-around time (TAT) from sample transmission to receiving decreased significantly from 51.5 (37.7, 73.0) min to 18.1 (14.4, 31.0) min ( U=0, P<0.001). After adopting the automatic sampling system, the TAT from receiving to sampling decreased from 41.0 (38.0, 45.0) min to 10.0 (5.0, 11.0) min ( U=0, P<0.001). The introduction of automatic quality control advanced the median time for the first batch of samples entering the biochemical testing pipeline from 9∶03 to 8∶27 and increased the reporting rate within 3 hours from 27.71%±1.39% to 36.90%±2.30% ( t=3.423, P<0.001). The intelligent monitoring system enabled module positioning monitoring, sample turn-around time reminder, instrument load rate monitoring, remaining reagent monitoring, and patient-based real-time quality control, resulting in improved instrument running efficiency and result accuracy. Followed by the introduction of the automatic audit function, the overall pass rate was 28.19%(10 006/35 500), including 37.17% (7 738/20 818) for biochemical reports, 31.57% (1 251/3 963) for chemiluminescence reports, and 9.49% (1 017/10 719) for biochemical immunity reports. The laboratory TAT decreased from (207.3±6.0) min to (169.8±5.9) min ( t=4.426, P<0.001). After the implementation of the critical value monitoring system, the timely reporting rate reached 99.52% (99.32%, 99.89%). After using quality digital management, outpatient biochemical immunity process was optimized to a decrease in laboratory TAT from 222 (201, 233) min to 145 (119, 195) min ( U=0, P=0.004), while the pass rate increased from 86.88% (85.91%, 87.81%) to 96.32% (95.86%, 96.96%) ( U=0, P=0.004). Conclusion:The establishment of an intelligent laboratory can optimize workflow, significantly improve the work efficiency and accuracy of sample processing, and minimize error.

Objective:To investigate the application value of intelligent laboratory construction in the laboratory.Methods:All samples sent to the Department of Clinical Laboratory from all the clinical departments at the Eighth Affiliated Hospital of Sun Yat-Sen University from January 2018 to June 2024 were collected. Full-process intelligentization building was achieved by the use of logistics transmission system, automatic sampling system, automatic quality control system, automatic audit system, intelligent monitoring system, critical value monitoring system, and intelligent DxlabReport operation management system, et al. The normal distribution data were analyzed by two independent samples t-test, and the skew distribution data were analyzed by Mann-Whitney U test. Results:After the implementation of the pneumatic logistics transmission system, the turn-around time (TAT) from sample transmission to receiving decreased significantly from 51.5 (37.7, 73.0) min to 18.1 (14.4, 31.0) min ( U=0, P<0.001). After adopting the automatic sampling system, the TAT from receiving to sampling decreased from 41.0 (38.0, 45.0) min to 10.0 (5.0, 11.0) min ( U=0, P<0.001). The introduction of automatic quality control advanced the median time for the first batch of samples entering the biochemical testing pipeline from 9∶03 to 8∶27 and increased the reporting rate within 3 hours from 27.71%±1.39% to 36.90%±2.30% ( t=3.423, P<0.001). The intelligent monitoring system enabled module positioning monitoring, sample turn-around time reminder, instrument load rate monitoring, remaining reagent monitoring, and patient-based real-time quality control, resulting in improved instrument running efficiency and result accuracy. Followed by the introduction of the automatic audit function, the overall pass rate was 28.19%(10 006/35 500), including 37.17% (7 738/20 818) for biochemical reports, 31.57% (1 251/3 963) for chemiluminescence reports, and 9.49% (1 017/10 719) for biochemical immunity reports. The laboratory TAT decreased from (207.3±6.0) min to (169.8±5.9) min ( t=4.426, P<0.001). After the implementation of the critical value monitoring system, the timely reporting rate reached 99.52% (99.32%, 99.89%). After using quality digital management, outpatient biochemical immunity process was optimized to a decrease in laboratory TAT from 222 (201, 233) min to 145 (119, 195) min ( U=0, P=0.004), while the pass rate increased from 86.88% (85.91%, 87.81%) to 96.32% (95.86%, 96.96%) ( U=0, P=0.004). Conclusion:The establishment of an intelligent laboratory can optimize workflow, significantly improve the work efficiency and accuracy of sample processing, and minimize error.

Objective:To investigate the effect and molecular mechanism of circular RNA-UBXN7 (circ_UBXN7) on the proliferation, migration and apoptosis of hepatocellular carcinoma cells.Methods:Circ_UBXN7 expression in the tissues and cells of hepatocellular cancer was detected by quantitative real-time polymerase chain reaction (qRT-PCR), and the relationship between circ_UBXN7 expression and clinicopathological features, including age, gender, tumor volume, pathological classification, staging, and lymph node metastasis was analyzed. The full-length sequence of circ_UBXN7 with lentivirus carrying lenti circ_UBXN7 and lenti circ_UBXN7 shRNA was constructed to transfect hepatocellular cell lines (HepG2 and Huh-7), respectively. CCK-8 experiments were performed to detect the ability of up- or down-regulation of circ_UBXN7 on the proliferation of HEPG2 and HUH-7 cells. Annexin V / PI experiment was used to detect the changes in apoptosis of HEPG2 and HUH-7 cells after up-regulation or down-regulation of circ_UBXN7 expression. JC-1 assay was used to detect the changes in mitochondrial potential energy of HEPG2 and HUH-7 cells after up-regulation or down-regulation of circ_UBXN7 expression. Transwell was used to detect the migration ability of HEPG2 and HUH-7 cells after up-regulation or down-regulation of circ_UBXN7 expression. Western blotting was used to detect the expressional change of TWIST, E-cadherin, N-cadherin and vimentin. Statistical analysis: The expression levels of circ_UBXN7 and clinicopathological features were measured by chi-square test. Two groups were compared by t-test and three groups and above were compared by single factor analysis of variance. LSD method was used for comparison between groups.Results:The expression of circ_UBXN7 in liver cancer tissues was significantly higher than adjacent tissues, and its expression level was significantly positively correlated with tumor volume, stage, and lymph node metastasis ( P < 0.05). Lenti-circ_UBXN7 had up-regulated the expression of circ_UBXN7 in HEPG2 and HUH-7 cells and promoted cell proliferation. Lenti-circ_UBXN7-shRNA had down-regulated the expression of circ_UBXN7 and induced apoptosis. Lenti-circ_UBXN7-shRNA had reduced the mitochondrial membrane potential of cells. Lenti-circ_UBXN7 had promoted cell migration, while lenti-circ_UBXN7-shRNA had inhibited cell migration. Lenti-circ_UBXN7 had induced increased expression of Twist, N-cadherin, and Vimentin proteins, and reduced the expression of E-cadherin protein. Lenti-circ_UBXN7-shRNA had opposite effects on the expression levels of each protein. Starbase V2.0 software showed that miR-203a and circ_UBXN7 had potential binding sites, and miR-203a and circ_UBXN7 expression levels were negatively correlated in HEP ??G2 and HUH-7 cells. Conclusion:circ_UBXN7 plays an important role in promoting the occurrence and development of liver cancer, and is expected to become a potential target for the treatment of liver cancer.

Objective:To explore the therapeutic effect of group cognitive-behavioral therapy (GCBT) for obsessive-compulsive disorder (OCD).Methods:This study used a randomized controlled trial design to compare GCBT with routine medication treatment. Unmedicated ninety-four patients who met the inclusion criteria were recruited and randomly allocated to GCBT group ( n=47) and drug treatment group ( n=47) by a simple random grouping method using the RAND function in Excel software which generated a table of random numbers to form a random grouping sequence. Both groups were treated for 12 weeks. The average reduction rate and value of Y-BOCS, HAMA 14 and HAMD 24 were compared between the two groups, t-test,chi-square (χ 2) test and variance analysis (ANOVA) were condulted to analyze data. Results:(1) There was no significant difference between two groups in Y-BOCS and HAMA 14 scores at baseline ( t=0.281, P=0.779; t=0.795, P=0.429), but HAMD 24 scores were significantly different ( t=2.316, P<0.05). Sixteen patients in GCBT group and sixteen in drug treatment group dropped out of treatment, resulted a total drop-out rate of 34%. There was no significant difference in the drop-out rate between the two groups. (2) After 12-week treatment, the Y-BOCS scores decreased compared to pre-treatment in both groups. There was no statistical difference in the mean reduction rate ((37.0±27.4)% vs. (45.5±22.9)%) and score (9.0±6.3 vs.11.0±5.8) of Y-BOCS ( F(1,62)=0.069, P=0.794; F(1,62)=0.001, P=0.975) before and after treatment between the two groups. There was no statistical difference in the effective and cure rate between the two groups (χ 2=1.653, P=0.199; χ 2=0.088, P=0.767) . (3) There was no significant difference in the mean reduction rate and score of HAMA 14 ( t=-0.922, P=0.362; t=1.082, P=0.286). (4) No significant difference was found regarding the mean reduction rate of HAMD 24 between the two groups, but the mean reduction scores of HAMD 24 in the medication group were significantly higher than those in GCBT group ( t=2.239, P=0.029). Conclusion:GCBT is equivalent to conventional medication treatment for obsessive-compulsive and anxiety symptoms for OCD patients, and medication treatment is superior to GCBT in depressive symptoms.

Objective:To explore the therapeutic effect of group cognitive-behavioral therapy (GCBT) for obsessive-compulsive disorder (OCD).Methods:This study used a randomized controlled trial design to compare GCBT with routine medication treatment. Unmedicated ninety-four patients who met the inclusion criteria were recruited and randomly allocated to GCBT group ( n=47) and drug treatment group ( n=47) by a simple random grouping method using the RAND function in Excel software which generated a table of random numbers to form a random grouping sequence. Both groups were treated for 12 weeks. The average reduction rate and value of Y-BOCS, HAMA 14 and HAMD 24 were compared between the two groups, t-test,chi-square (χ 2) test and variance analysis (ANOVA) were condulted to analyze data. Results:(1) There was no significant difference between two groups in Y-BOCS and HAMA 14 scores at baseline ( t=0.281, P=0.779; t=0.795, P=0.429), but HAMD 24 scores were significantly different ( t=2.316, P<0.05). Sixteen patients in GCBT group and sixteen in drug treatment group dropped out of treatment, resulted a total drop-out rate of 34%. There was no significant difference in the drop-out rate between the two groups. (2) After 12-week treatment, the Y-BOCS scores decreased compared to pre-treatment in both groups. There was no statistical difference in the mean reduction rate ((37.0±27.4)% vs. (45.5±22.9)%) and score (9.0±6.3 vs.11.0±5.8) of Y-BOCS ( F(1,62)=0.069, P=0.794; F(1,62)=0.001, P=0.975) before and after treatment between the two groups. There was no statistical difference in the effective and cure rate between the two groups (χ 2=1.653, P=0.199; χ 2=0.088, P=0.767) . (3) There was no significant difference in the mean reduction rate and score of HAMA 14 ( t=-0.922, P=0.362; t=1.082, P=0.286). (4) No significant difference was found regarding the mean reduction rate of HAMD 24 between the two groups, but the mean reduction scores of HAMD 24 in the medication group were significantly higher than those in GCBT group ( t=2.239, P=0.029). Conclusion:GCBT is equivalent to conventional medication treatment for obsessive-compulsive and anxiety symptoms for OCD patients, and medication treatment is superior to GCBT in depressive symptoms.

To investigate the potential risk factors for hepatocellular carcinoma in primary biliary cholangitis (PBC) patients. Methods:The data of 670 PBC inpatients between January 2011 and December 2016 were collected from the database of The First Affiliated Hospital of Zhengzhou University. The potential risk factors were evaluated, and odds ratios (ORs) and 95% confidence intervals (CIs) were analyzed by univariate (unadjusted OR) and multivariate [adjusted OR (AOR)] conditional Logistic regression. Results: In total, 35 PBC patients developed liver carcinoma (5.2%); of these, 4 patients (female) were excluded because of incomplete data for influencing factors and 6 (2 male; 4 female) were excluded as they were diagnosed with hepatocellular carcinoma (HCC) during or before PBC. Therefore, 25 patients were included in the case-control study. Male patients were more likely than female patients to show alcohol in-take, smoking, a family history of malignancy, and serious liver injury (all P<0.05), indicated by the increasing levels of alanine amino-transferase (ALT), aspartate aminotransferase (AST), and gamma glutamyl transferase (GGT) (P<0.05). Conditional Logistic regression analysis revealed that body mass index (BMI) ≥25 kg/m2 (AOR=1.015, 95% CI: 1.001-1.257, P=0.032) and history of alcohol intake (AOR=10.014, 95% CI: 1.009-91.071, P=0.039) were significantly associated with increased odds of HCC development in PBC patients. Conclusions:The risk factors for PBC-associated liver carcinoma include BMI≥25 kg/m2 and history of alcohol intake. In addition to regular monitoring, PBC patients may benefit from alcohol abstinence and body weight control.

Objective To investigate the role and mechanism of circular RNA-vimentin (circ-VIM) in the proliferation and apoptosis of colorectal cancer cells.Methods From December 2016 to December 2017, at Department of General Surgery of The First Affiliated Hospital of Zhengzhou University, the clinical data of 100 patients who underwent radical resection of colorectal cancer and were confirmed by pathological examination after operation were collected.The tumor tissues and corresponding paracancerous tissues (negative control) were also collected.The expression of circ-VIM in the colorectal cancer tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR).The proliferation of HCT-116 and HT29 colorectal cancer cells was detected by cell counting kit-8 assay.The ratio of apoptosis of HCT-116 and HT29 cells was measured by annexin Ⅴ/propidium iodide double staining assay.The mitochondrial membrane potential of HCT-116 and HT29 cells was examined by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1) assay.The expression changes of protein kinase B and mammalian target of rapamycin were tested by Western blotting.The target miRNA of circ-VIM was predicted by miRDB software.T-test and chi-square test were performed for statistical analysis. Results The expression of circ-VIM in colorectal cancer tissues was 2.387 ±0.536, which was higher than that in corresponding paracancerous tissues (1.110 ±0.134), and the difference was statistically significant (t =23.096, P <0.01).And the expression levels of circ-VIM were significantly different in patients with different tumor size, TNM stage and lymph node metastasis (all P <0.05).The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM group was 0.737 ±0.023 and 0.835 ±0.025, respectively, which were both higher than those in control group (0.449 ±0.020 and 0.531 ±0.019), and the differences were statistically significant (t =20.706 and-15.374, both P <0.01).The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 0.236 ±0.027 and 0.243 ±0.019, which were lower than those in control group, and the differences were statistically significant (t =24.557 and -23.197, both P <0.01).The ratio of apoptosis of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was (18.00 ±1.82)％ and (20.80 ±0.61)％, which was higher than those in control group ((6.64 ±2.01)％ and (7.35 ±1.36)％), and the differences were statistically significant (t =8.826 and 17.454, both P <0.01).The fluorescence intensity ratio of JC-1 aggregate and JC-1 monomer of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 2.21 ±0.12 and 1.40 ±0.11, which was lower than those in control group (14.54 ±1.00 and 9.24 ±1.18), and the differences were statistically significant (t =-19.558 and-15.685, both P <0.01), which indicated mitochondrial membrane potential decreased.After treated with lenti-circ-VIM-shRNA, the expression of phosphorylated protein kinase B, phosphorylated mammalian target of rapamycin, B-cell lymphoma-2 and mitochondrial cytochrome C at protein level were all down-regulated, however the expression of cytoplasmic cytochrome C, B-cell lymphoma-2 associated X protein and cleaved caspase-3 at protein level were all up-regulated.When the expression of circ-VIM was up-regulated, the expression of miRNA-147b, miRNA-4447 and miRNA-3656 was down-regulated.When the expression of circ-VIM was down-regulated, the expression of miRNA-147b, miRNA-4447 and miRNA-3656 was up-regulated.Conclusion The expression of circ-VIM in colorectal cancer is abnormally increased, which is involved in the proliferation and apoptosis of colorectal cancer cells.

Objective To investigate the identification of staphylococcus aureus lineage ST59 using the combined detection of delta hemolysin allelic variant G10S(HldG10S) and beta hemolysin(β-toxin). Methods Perspective study.A total of 82 non-duplicate clinical staphylococcus aureus were collected from November 2017 to April 2018 in the department of Clinical laboratory, the Third Xiangya Hospital of Central South University, China.The strains were routinely identified by MALDI-TOF MS and the mass spectra were obtained. According to the m/z expression intensity of delta hemolysin(Hld), all strains were divided into three groups:HldG10S (3036±2.0)m/z, Hld (3006±2.0)m/z and ND [no (3036±2.0)m/z and no (3006±2.0)m/z]. The distribution of ST59 in the three groups was detected by MLST. Reverse synergic hemolysis test was used to determine theβ-toxin phenotype. And the sensitivity, specificity and accuracy of HldG10S,β-toxin and the combined detection of HldG10S and Hld to identify ST59 were compared. Results Among the 82 strains, 21 strains expressed HldG10S toxin, accounting for 25.6％. 39 strains expressed Hld toxin, accounting for 47.6％.22 strains did not express HldG10S and Hld toxin, accounting for 26.8％. In HldG10S group,16 strains were ST59, accounting for 76.19％(16/21).ST59 was not found in both Hld and ND groups. All 16 strains of ST59 in HldG10S group producedβ-toxin, while none of the 5 strains of non-ST59 producedβ-toxin. The specificity(100％) and accuracy(100％) of the combined detection was significantly higher than that of HldG10S andβ-toxin single detection of specificity(92.4％, 77.3％) and accuracy(80.5％, 81.7％) (χ2=19.472, P<0.001;χ2=17.792, P<0.001). Conclusion The combined detection of HldG10S andβ-toxin can preliminarily and rapidly identify ST59, which can assist the routine monitoring of the change trend of staphylococcus aureus epidemic.

Objective To investigate the effects of 3 kinds of broth media,including tryptose soya broth(TSB),LB and M-H,on the biofilm formation of Staphylococcus epidermidis (S.epidermidis).Methods The effects of TSB,LB and M-H broth media on the biofilm formation of S.epidermidis were investigated by the construction of bacterial biofilm in 96-well and 6-well microplates and the crystal violet straining for the semi-quantitative analysis and microscopic observation of the bacterial biofilm.The effects of TSB,LB and M-H broth media on the expression of adhesion-related genes in S.epidermidis were determined by the extraction of bacterial total RNA,reverse transcription and real-time PCR.Results Compared with LB (0.149 ± 0.047) and M-H (0.323 ± 0.003) media,TSB medium (2.954 ± 0.287) could significantly promote the biofilm formation of S.epidermidis (TSB vs LB,t =16.706,P ＜ 0.01;TSB vs M-H,t =15.877,P ＜ 0.01).Compared with LB medium,TSB medium could significantly enhance the expression of icaA gene (t =9.667,P＜0.01) but inhibit icaR gene (t =13.283,P＜0.01).Conclusion Compared with LB and M-H broth media,TSB medium may significantly improve the biofilm formation and the expression of adhesion-related gene icaA of S.epidermidis.