AIM:To study the blocking effect of shRNA on the expression of PSMA gene in LNCaP cell line by using shRNA eukaryotic expression vector.METHODS:Three pairs of DNA templates coding shRNA,synthesized against PSMA and cloned into the vector pSilencer 2.1-U6-neo,which was named pSilencer 2.1-U6-neo-shRNA,were identified by restriction endonuclease digestion analysis and DNA sequencing.LNCaP cells were then transfected with these three pSilencer 2.1-U6-neo-shRNAs and the negative control pSilencer 2.1-U6-neo-NC.After G418 selection,the cells were selected and the interfering effect was detected by RT-PCR and Western blotting.The biological behaviours of the transfected LNCaP cells were also tested.RESULTS:Restriction endonuclease digestion analysis and DNA sequencing results all showed that the 3 target segments were cloned into pSilencer 2.1-U6-neo vector respectively.After transfected into LNCaP cells,the inhibitory ratio of PSMA mRNA was 33.15%,9.26% and 41.97% respectively,and that of PSMA protein was 26.26%,6.47%,40.69% respectively.The p-shRNA3 was chosen to test the cell growth and its invasive power in vitro.The results showed that after interfering,the invasiveness of LNCaP cells were enhanced.CONCLUSION:The vector-based shRNA on PSMA gene effectively knocks down the PSMA gene expression.The successful construction of PSMA shRNA makes it possible for further study of the interaction between PSMA and prostate cancer.

Objective To discuss the standard of endoscopic diagnosis of duodenal white spot syndrome and to test its accuracy by pathological diagnosis standard. Methods A total of 6,995 patients undergoing gastroscopy were detailedly examined through gastrofiberscope or electrogastroscope and endoscopic diagnosis were made by the standard of endoscopic diagnosis of duodenal white spot syndrome. Then, pathological examination was done to test and verify the accuracy of the standard of endoscopic diagnosis. Results There were 533 patients suffering from duodenal white spot pathological changes, accounting for 7.62%. And duodenal mucositis was confirmed by pathological findings in all 90 cases of the patients undergone their biopsy at the same time. Conclusions The appropriate standard of endoscopic diagnosis can improve the detection rate of duodenal white spot syndrome and there is a high coincidence rate between endoscopic diagnosis and pathological diagnosis.

AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.

Adult ; Brachydactyly ; genetics ; Chromosome Aberrations ; Female ; Fingers ; abnormalities ; Humans ; Male ; Pedigree

Objective To investigate the characteristics of clinical and cardiovascular magnetic resonance imaging(CMR) of arrhythmogenic cardiomyopathy with left ventricular(ALVC) involvement. Methods Ten cases of arrhythmogenic cardiomyopathy with left ventricular involvement and sixteen randomly chosen cases of arrhythmogenic cardiomyopathy with right ventricular (ARVC) involvement were enrolled in this study. Clinical symptoms, cardiac electrophysiological changes as well as the cardiac morpharage, ventricular functions and delayed enhancement of myocardium assessed by CMR were compared between the two groups. The size of heart chambers, global ventricular functions and the fat/fibrosis infiltration were evaluated by turbo fast spin echo, ture FISP cine and delayed enhancement. These were statistically analyzed by independent samples t test, respectively.Results There was no significant difference in age and gender between two groups. All the patients in two groups presented non-specific clinical manifestations with no significant differences (P>0.05) except for short of breath(P=0.034). The end-diastolic diameter of left ventricular, left ventricular end-diastolic volume index, left and right ejection fraction in ALVC and ARVC group were (64.2±7.7), (49.2±5.9) mm(t=5.551,P<0.001), (113.9±24.0), (69.2± 30.0) ml/m2(t=3.962, P<0.001), (38.5±3.1)%, (56.0±8.4)%(t=-6.733,P<0.001), (42.0±5.5)%, (18.0±7.3)%(t=8.817, P<0.001) respectively. An medium of 11 and 14 segments of fat or fat/fibrosis were found in intramural wall of the LV myocardiumin patients with ALVC,while only 0 segment(all P<0.001)was found in patients with ARVC.Conclusions The clinical manifestations of arrhythmogenic cardiomyopathy with left ventricular involvement was similar to ARVC, however, with regarding to cardiac morphological, functional and myocardial lesions, these two diseases have different characteristics in CMR features.“one-stop-shop”MRI examination has high value in the diagnosis of arrhythmogenic cardiomyopathy with Left ventricular involvement.

ObjectiveTo investigate the change in the activity of glucosylceramide synthase, the key enzyme in glycosphingolipid metabolism and synthesis, in Huh7 cells infected by hepatitis B virus (HBV) in vitro. MethodsBlood samples were collected from nine previously untreated patients with acute hepatitis B who attended Department of Infectious Diseases, The First Hospital of Lanzhou University, from June to August, 2019, and the blood samples collected from seven healthy individuals who underwent physical examination were established as control. Huh7 cells were inoculated with the high-copy HBV particles (＞9.9×107 IU/ml) in the serum of patients with HBV infection (infection group), and Huh7 cells co-cultured with the serum of healthy individuals were established as control group. The expression levels of HBsAg and HBV DNA in the cytoplasm of HBV-infected Huh7 cells were measured, and the correlation between GCS activity and virus was analyzed. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups, and a Pearson correlation analysis was performed. ResultsCompared with the control group, the infection group had a significant reduction in the number of cells, an increase in cell volume, and cell membrane fragmentation. The infection group had a significant increase in the expression of HBsAg in cytoplasm at 4 hours, 8 hours, 2 days, and 5 days after infection (P＜0.05); the expression level of HBV DNA tended to increase significantly from 4 hours after infection to 8 hours, 2 days, and 5 days after infection (16.67±11.55 IU/ml vs 112.01±25.94 IU/ml/328.01±10350 IU/ml/101.60±49.84 IU/ml, P＜0.001), with the highest level at 2 days after infection. During HBV infection, the activity of GCS gradually increased with the increase in viral replication from 4 hours after infection (126.21±9.59 IU/ml) and reached a peak at 2 days after infection (226.53±36.27 IU/ml), with a significant difference between the infection group and the control group at 2 days after infection (226.53±36.27 IU/ml vs 136.50±1544 IU/ml, t=3.956, P=0.016 7). The activity of GCS was positively correlated with HBV DNA level (r=0.576 8, P=0047 1). ConclusionHuh7 cells are successfully infected with the high-copy HBV particles in the serum of patients with HBV infection, which mimics the characteristics of HBV infection in vitro to a certain degree. The activity of GCS may be associated with HBV infection, suggesting that glycosphingolipid synthesis and metabolism may be closely associated with HBV.

Objective:To investigate the targeted surveillance combined with cluster management on isolation protector use rate and hand hygiene qualification rate in caregivers of patients with respiratory tract multidrug-resistant bacterial infections.Methods:Second Hospital of Shanxi Medical University began to perform targeted surveillance combined with cluster management among caregivers of patients with multidrug-resistant bacterial infections in July 2019. The caregivers of 50 patients with respiratory tract multidrug-resistant bacterial infections who were admitted before targeted surveillance combined with cluster management were included in the pre-implementation group. The caregivers of 50 patients with respiratory tract multidrug-resistant bacterial infections who were admitted after targeted surveillance combined with cluster management were included in the post-implementation group. Multidrug-resistant bacterial infections in caregivers, isolation protector use rate, environmental disinfection and hand hygiene qualification rate were compared between before and after 6 months of targeted surveillance combined with cluster management. The application value of targeted surveillance combined with cluster management in the prevention of respiratory tract multidrug-resistant bacterial infections was analyzed.Results:The number of caregivers with respiratory tract multidrug-resistant bacterial infections in the post-implementation group was lower than that in the pre-implementation group (5 vs. 13, P < 0.05). The proportion of caregivers who wear a mask (94.11%), isolation clothes (80.39%) and gloves (98.03%) in the post-implementation group was significantly higher than that in the pre-implementation group (70.00%, 62.00%, 78.00%, χ2 = 10.027, 4.911, 9.683, all P < 0.05). The proportion of caregivers who were qualified in terms of hand hygiene (82.35%), object surface (76.47%), and keeping the air fresh in the post-implementation group was significantly higher than that in the pre-implementation group (64.00%, 58.00%, 64.00%, χ2 = 4.341, 3.915, 5.450, all P < 0.05). Conclusion:Targeted surveillance combined with cluster management can effectively help prevent respiratory tract multidrug-resistant bacterial infections in caregivers and increase isolation protector use rate and hand hygiene qualification rate.

Objective: To explore the effectiveness of machine learning algorithms in predicting non-suicidal self-injury (NSSI) behavior among college students, and to analyze the influencing factors of NSSI behavior, thus providing a reference for promoting psychological well-being. Methods: In December 2022, a stratified random cluster sampling method was used to select 835 college students from a university in Guizhou Province, China. The Adolescent Self-injury Scale, Family Function Assessment Scale, and Emotion Regulation Self-efficacy Scale were used to evaluate the participants. Demographic characteristics, family factors, and emotional factors were taken as independent variables, while the dependent variable was whether college students exhibited NSSI behavior. Machine learning algorithms, including Logistic regression, support vector machine (SVM), decision trees, algorithm gradient boosting trees, random forests, and AdaBoost, were used to construct predictive models. Results: The detection rate of NSSI behavior among the college students was 23.23% (194 individuals). The NSSI behavior group scored higher than the non-NSSI behavior group in total family function, emotional communication, egoism, and family rules ( t=3.02, 3.35 , 2.23,2.87, P <0.05). On the other hand, the non-NSSI behavior group scored higher than the NSSI behavior group in total emotion regulation selfefficacy, managing negative emotion self-efficacy, and expressing positive emotion self-efficacy ( t=-5.04, -5.48 , -2.43, P <0.05). The recall rates of random forests, SVM, Logistic regression, decision trees, algorithm gradient boosting trees, and AdaBoost were 84.3% , 90.6%, 73.4%, 87.5%, 95.3%, 89.0%, respectively. The F1 scores were 84.4%, 92.1%, 71.2 %, 79.4%, 91.7%, 89.1% , respectively. The respective precision rates were 84.4%, 93.5%, 69.1%, 72.7%, 88.4%, 89.1 %. The AUC scores were 0.845, 0.922, 0.706, 0.776, 0.915, and 0.891, respectively. Conclusion Compared to the algorithm gradient boosting tree, random forest, Logistic regression, and AdaBoost models, the SVM model has a better predictive effect on whether college students in Guizhou Province exhibits NSSI behavior. It is recommended to use an appropriate model to identify students at risk of NSSI behavior as early as possible and provide psychological crisis interventions to promote their mental health.

Objective To investigate the effect of the overexpression of Krüppel-like factor 6 (KLF6)towards the apoptosis of human lens epithelial cells (HLECs) induced by ultraviolet B (UVB) radiation.Methods The eukaryotic expression plasmid pEGFP-C2-KLF6 which was successfully constructed were transfected into HLECs,followed by the detection of KLF6 level by using Western blot,and then companied by UVB stimulation.Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay.The morphology of the cells was observed by using hematoxylin-eosin staining method.The cell damage was examined by Live/Dead staining.The apoptotic markers bax and bcl-2 were detected by Western blot.Quantitative apoptotic levels were measured with the apoptosis detection kit;the expression level of reactive oxygen species (ROS) was analyzed by DCFH-DA probe.Results The cell viability of the 0.5 μg transfection group and the 1.0 μg transfection group was significantly lower than that of the blank vector control group (both at P＜0.05).In high KLF6 expression group,the cells were sparse,long and narrow in size and shape,and the cytoplasm was concentrated.The cells in the normal control group were green living cells with stable morphology and even quantity.The number of red dead cells was increased significantly in the KLF6 highexpression group.After UVB irradiation,the apoptosis value,relative bax expression,bax/bcl-2 ratio and ROS expression of HLECs cells in the KLF6 high-expression group were all higher than those in the blank vector control group,with statistically significant differences between them (all at P＜0.05).Conclusions Overexpression of KLF6 can exacerbate apoptosis of HLECs caused by UVB,by regulating the expression of apoptosis-related proteins and promoting the accumulation of ROS in the endoplasmic reticulum.Down-regulation of KLF6 expression by biological tools may play a protective role on LECs to a certain extent.

Objective To investigate the regulating effects of Krüppel-like factor 6 (KLF6) on the apoptosis of human lens epithelial cells (HLECs) by activating transcription factor 4 (ATF4) pathway and explore the bio-molecular mechanism of KLF6/ATF4-induced HLECs apoptosis.Methods HLECs (HLE-B3) were cultured using high glucose DMEM medium.The eukaryotic expression plasmid pEGFP-C2-ATF4 was transfected into the cells by liposome 2000 in the ATF4-transfected group,and pEGFP-C2 was transfected in the empty plasmid group.Then the cells were exposed to 20 mJ/cm2 ultraviolet ray B (UVB) for 200 seconds,The morphological changes of the cells were observed by hematoxylin & eosin staining and Hoechst33258 fluorescein staining.Cultured cells were transfected using pEGFP-C2-KLF6 and pEGFP-C2 plasmid and pSilencer-KLF6 (siKLF6) and pSilencer plasmid,respectively,and the expression of ATF4 protein in the cells was detected by Western blot assay.Culture cells were divided into four groups.pEGFP-C2 and pSilencer plasmids were co-transfected into the cells in the empty plasmid group;pEGFP-C2-KLF6 and pSilencer empty plasmid were co-transfected into the cells of the KLF6 + pSilencer group;pEGFP-C2 empty plasmid and pSilencer-ATF4 were co-transfected in the cells of the siATF4 + pEGFP-C2 group;pEGFP-C2-KLF6 and pSilencer-ATF4 plasmids were co-transfected in the cells of the KLF6 + siATF4 group,and then the cells were exposed to UVB.The apoptosis of the cells were detected by ELISA assay.Results Cultured cells grew well in the normal control group with the uniform morphology and regular arrangement.The karyopyknosis,karyorrhexis and enlargement of intercellular space were found in the cells exposed to UVB.In the ATF4 transfected group,the number of cells was decreased.The relative expression level of the ATF4 protein in the cells was 0.99±0.06 and 0.13±0.02 in the UVB+ATF4 transfected group and UVB+pEGFP-C2 plasmid group,respectively,with a significant difference between them (t =23.13,P＜0.01).The relative expression levels of KLF6 and ATF4 proteins in the KLF6 transfected group were higher than those in the empty plasmid group,and the relative expression levels of KLF6 and ATF4 proteins in the siKLF6 group were significantly lower than those in the empty plasmid group (all at P＜0.01).ELISA assay showed that the apoptotic rate in the ATF4 transfected group was 1.37± 0.11,which was significantly higher than 0.31 ±0.11 in the normal control group (t =8.034,P =0.001);the apoptotic rate of the cells was increased in the KLF6+pSilencer group and decreased in the siATF4+pEGFP-C2 group in comparison with the empty plasmid group (P＜0.01,P=0.02).In addition,the apoptotic rate in the KLF6+ siATF4 group was remarkably lower than that in the KLF6 + pSilencer group (P＜ 0.01).Conclusions KLF6 promotes the apoptosis of HLECs induced by UVB radiation.Silence of ATF4 gene reduces the apoptotic rate of the cells.ATF4 is probably a target factor in the regulating oathwav of KLF6 to apoptosis.