Objective:To explore the role of circ_0007766 molecule cyclized from the erb-b2 receptor tyrosine kinase 2(ERBB2)gene in prostate cancer(PCa)cells and the impact of hypoxia on the regulation of its expression.Methods:The expression of circ_0007766 molecule in PCa cells and tissues was detected by qRT-PCR.The siRNA of circ0007766 was transfected to PCa cells(DU145 cell and PC3 cell)respectively;Colony formation assay,CCK-8 assay and Transwell assay were performed to detect the effect of circ_0007766 knockdown on the colony formation,proliferation,migration,and invasion abilities of PCa cells.Hypoxia cell model of DU145 and PC3 cells were established by hypoxic chamber method,and WB was performed to detect the protein expression of HIF-1α,a key molecule of the hypoxic pathway.qRT-PCR was performed to detect the expression of circ_0007766 molecule in hypoxic cell model,and the protein expression of RNA-binding protein eukaryotic translation initiation factor 4A3(EIF4A3)was detected by WB.RNA immunoprecipitation(RIP)was performed to detect the binding of EIF4A3 to circ_0007766 under hypoxic conditions.qRT-PCR assay was performed to further detect the effect of EIF4A3 knockdown on the expression of circ_0007766 under hypoxic conditions.Results:circ_0007766 was highly expressed in PCa cells(P<0.01)and PCa tissues(P<0.05).The knockdown of circ_0007766(P<0.05 or P<0.01)could significantly inhibit the proliferation,migration and invasion abilities of PCa cells(all P<0.01).The upregulation of HIF-1α protein under hypoxic conditions confirmed the successful establishment of the hypoxia cell model.qRT-PCR analysis revealed that compared with that in the normoxia group,circ_0007766 expression was markedly elevated in the hypoxia group(P<0.01).WB analysis demonstrated increased EIF4A3 protein expression in cells of the hypoxia group.RIP assays indicated that circ_0007766 was highly concentrated in the EIF4A3-enriched group(P<0.01).Additionally,qRT-PCR showed that hypoxia significantly boosted circ_0007766 expression,whereas EIF4A3 knockdown notably diminished the hypoxia-induced expression of circ_0007766(P<0.05 or P<0.01).Conclusion:Circ-0007766 plays the role of cancer-promoting molecule in PCa cells,and its expression is related to the regulation of EIF4A3 molecule under hypoxic conditions.

Objective:To assess the effect of Hood's technique on urinary continence in patients undergoing single-port robot-assisted radical prostatectomy (spRARP).Methods:The clinical data of 53 patients who underwent spRARP performed by a single surgeon in the First Affiliated Hospital of Harbin Medical University from June 2021 to October 2023 were retrospectively analyzed. There were 25 patients in the spRARP+ Hood group and 28 patients in the spRARP group. There were no statistically significant differences between the spRARP+ Hood group and the spRARP group in terms of patients′ age [(70.28±5.98) years vs. (60.89±6.86) years old], body mass index[(24.64±2.85) kg/m 2 vs. (24.59±3.17) kg/m 2], prostate weight [70.00 (40.69, 102.25) g vs. 73.50 (49.13, 94.50) g], total prostate specific antigen[8.62 (4.56, 15.26) ng/ml vs. 12.68 (6.99, 19.24) ng/ml], Gleason score [8 (7, 8) vs. 8 (7, 8)], age-adjusted Charlson comorbidity index (aCCI) [4 (3, 5) vs. 3 (3, 4)], and clinical stage [T 2a/T 2b/T 2c: 6/10/9 cases vs. 5/7/16 cases ] ( P>0.05). In the SpRARP + Hood group, the detrusor apron, tendon arch, pubic prostatic ligament, and dorsal vascular complex were completely preserved during the operation. In contrast, this was not the case for spRARP.Additionally, the incision size, bleeding volume, intraoperative blood transfusion volume, operation time, gastrointestinal function recovery time, total amount of drainage in the first three postoperative days, retention time of the drainage tube, postoperative hospitalization time, positive incision margins, recovery rate of urinary continence immediately after removal of the urinary catheter, and the recovery rate of urinary continence at 1, 3, and 6 months postoperatively were compared between the two groups. Results:All 53 surgeries were successfully completed. The differences in incision length [4.0 (3.5, 4.0) cm vs. 4.0 (4.0, 4.0) cm], intraoperative bleeding [50 (40, 100) ml vs. 100 (50, 100) ml], and intraoperative transfusion rate [4.0% (1/25) vs. 17.8% (5/28)] were not statistically significant between the spRARP+ Hood group and the spRARP group ( P>0.05), and the difference in operative time [205.0 (167.5, 240.0) min vs. 242.5 (185.0, 300.0) min] was statistically significant( P<0.05).The recovery time of gastrointestinal function in the spRARP+ Hood group vs. the spRARP group [1.0 (1.0, 1.5) d vs. 1.0 (1.0, 2.0) d], total amount of drainage in the first 3 d postoperatively [150.00 (72.50, 261.00) ml vs. 230.00 (115.00, 417.50) ml], duration of drain retention [5.0 ( 4.0, 5.0) d vs. 5.0 (4.0, 6.8) d], postoperative hospital stay [5.0 (4.0, 7.5) d vs. 5.0 (3.3, 7.8) d], and margin positivity rate [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). The postoperative Clavien-Dindo complication classification was grade I in both groups. The differences between the spRARP+ Hood group and the spRARP group in the rates of recovery of urinary continence immediately after the urinary catheter removal [56.0% (14/25) vs. 7.1% (2/28)] and one month after surgery [76.0% (19/25) vs. 28.5% (8/28)] were statistically significant ( P<0.05). The differences in the rates of recovery of urinary continence at 3 months after surgery [80.0% (20/25) vs. 67.8% (19/28)], at 6 months after surgery [88.0% (22/25) vs. 96.4% (27/28)], and biochemical recurrence at 6 months after surgery [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). Conclusions:The outcomes of spRARP+ Hood in the treatment of localized prostate cancer were comparable to those of spRARP. However, spRARP+ Hood has better urinary continence immediately after removal of the urinary catheter and 1 month postoperatively.

Objective To investigate the effect of the loss of myeloid-derived growth factor(Mydgf)on the transformation of cardiac fibroblasts into myofibroblasts after myocardial infarction(MI).Methods Two adult mouse groups,including a wild-type(WT)group and another group with Mydgf knockout(Mydgf-KO),were examined in the study.The mice in these two groups were tested for their cardiac function by measuring left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)(n=10).Quantitative real-time PCR(qRT-PCR)(n=3)was performed to determine the mRNA expression levels of myofibroblast markers,including α-smooth muscle actin(α-SMA),periostin(postn),type Ⅷ collagen(col8al),and connective tissue growth factor(ctgf).Western blot(n=3)was performed to verify the protein expression levels of α-SMA.MI modeling was performed on the WT and the Mydgf-KO mice.Postoperative LVEF and LVFS(n=10)were then measured.The hearts were harvested and Masson staining was performed to determine the infarcted area(n=10).The heart samples of Mydgf-KO and WT mice were collected at d 7 and d 14 after MI,respectively,to verify the expression of myofibroblast markers(n=3).Results Compared with WT mice,LVEF and LVFS in adult Mydgf-KO mice showed no significant changes(all P＞0.05).However,the mRNA levels of α-SMA and postn were upregulated,and α-SMA protein expression was also increased(all P＜0.05).After MI,compared with WT mice,LVEF and LVFS in Mydgf-KO mice decreased,and the infarcted area increased significantly(all P＜0.05).Furthermore,mRNA levels of α-SMA,col8al,postn,and ctgf were increased in Mydgf-KO mice.In addition,the α-SMA protein expression level was upregulated and α-SMA-positive fibroblasts were increased(P＜0.05).Conclusion Mydgf deletion promotes the transformation of cardiac fibroblasts into myofibroblasts and aggravates myocardial fibrosis after MI.

Objective:To assess the effect of Hood's technique on urinary continence in patients undergoing single-port robot-assisted radical prostatectomy (spRARP).Methods:The clinical data of 53 patients who underwent spRARP performed by a single surgeon in the First Affiliated Hospital of Harbin Medical University from June 2021 to October 2023 were retrospectively analyzed. There were 25 patients in the spRARP+ Hood group and 28 patients in the spRARP group. There were no statistically significant differences between the spRARP+ Hood group and the spRARP group in terms of patients′ age [(70.28±5.98) years vs. (60.89±6.86) years old], body mass index[(24.64±2.85) kg/m 2 vs. (24.59±3.17) kg/m 2], prostate weight [70.00 (40.69, 102.25) g vs. 73.50 (49.13, 94.50) g], total prostate specific antigen[8.62 (4.56, 15.26) ng/ml vs. 12.68 (6.99, 19.24) ng/ml], Gleason score [8 (7, 8) vs. 8 (7, 8)], age-adjusted Charlson comorbidity index (aCCI) [4 (3, 5) vs. 3 (3, 4)], and clinical stage [T 2a/T 2b/T 2c: 6/10/9 cases vs. 5/7/16 cases ] ( P>0.05). In the SpRARP + Hood group, the detrusor apron, tendon arch, pubic prostatic ligament, and dorsal vascular complex were completely preserved during the operation. In contrast, this was not the case for spRARP.Additionally, the incision size, bleeding volume, intraoperative blood transfusion volume, operation time, gastrointestinal function recovery time, total amount of drainage in the first three postoperative days, retention time of the drainage tube, postoperative hospitalization time, positive incision margins, recovery rate of urinary continence immediately after removal of the urinary catheter, and the recovery rate of urinary continence at 1, 3, and 6 months postoperatively were compared between the two groups. Results:All 53 surgeries were successfully completed. The differences in incision length [4.0 (3.5, 4.0) cm vs. 4.0 (4.0, 4.0) cm], intraoperative bleeding [50 (40, 100) ml vs. 100 (50, 100) ml], and intraoperative transfusion rate [4.0% (1/25) vs. 17.8% (5/28)] were not statistically significant between the spRARP+ Hood group and the spRARP group ( P>0.05), and the difference in operative time [205.0 (167.5, 240.0) min vs. 242.5 (185.0, 300.0) min] was statistically significant( P<0.05).The recovery time of gastrointestinal function in the spRARP+ Hood group vs. the spRARP group [1.0 (1.0, 1.5) d vs. 1.0 (1.0, 2.0) d], total amount of drainage in the first 3 d postoperatively [150.00 (72.50, 261.00) ml vs. 230.00 (115.00, 417.50) ml], duration of drain retention [5.0 ( 4.0, 5.0) d vs. 5.0 (4.0, 6.8) d], postoperative hospital stay [5.0 (4.0, 7.5) d vs. 5.0 (3.3, 7.8) d], and margin positivity rate [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). The postoperative Clavien-Dindo complication classification was grade I in both groups. The differences between the spRARP+ Hood group and the spRARP group in the rates of recovery of urinary continence immediately after the urinary catheter removal [56.0% (14/25) vs. 7.1% (2/28)] and one month after surgery [76.0% (19/25) vs. 28.5% (8/28)] were statistically significant ( P<0.05). The differences in the rates of recovery of urinary continence at 3 months after surgery [80.0% (20/25) vs. 67.8% (19/28)], at 6 months after surgery [88.0% (22/25) vs. 96.4% (27/28)], and biochemical recurrence at 6 months after surgery [4.0% (1/25) vs. 3.6% (1/28)] were not statistically significant ( P>0.05). Conclusions:The outcomes of spRARP+ Hood in the treatment of localized prostate cancer were comparable to those of spRARP. However, spRARP+ Hood has better urinary continence immediately after removal of the urinary catheter and 1 month postoperatively.

Objective:To investigate clinical characteristics of bullous pemphigoid (BP) developing after the treatment with dipeptidyl peptidase-Ⅳ inhibitors (DPP4i) in patients with diabetes mellitus.Methods:A total of 116 inpatients with BP complicated by diabetes mellitus were collected from the Seventh People′s Hospital of Shenyang between January 2014 and December 2020, and divided into 2 groups: DPP4i-BP group treated with DPP4i before the onset of BP, and general BP group receiving no treatment with DPP4i. General clinical data, skin lesion area, laboratory indicators, treatment regimens, and prognosis were analyzed and compared between the above 2 groups, the time interval from the administration of DPP4i to the diagnosis of BP was recorded in the DPP4i-BP group. One-way analysis of variance was used to compare measurement data among multiple groups, two-independent-sample t test was used for comparisons between two groups, and paired t-test for intra-group comparisons before and after treatment; chi-square test was used to compare enumeration data between groups. Results:There were 32 patients aged 77.17 ± 15.32 years in the DPP4i-BP group, with a male-to-female ratio being 15∶17; there were 84 patients aged 76.65 ± 19.32 years in the general BP group, with a male-to-female ratio being 43∶41. The time interval from the administration of DPP4i to the diagnosis of BP was 14.61 ± 3.93 months in the DPP4i-BP group. The time interval for vildagliptin was the shortest (5.42 ± 2.84 months) , and there was a significant difference in the time interval among vildagliptin, sitagliptin, linagliptin and saxagliptin ( F= 8.93, P < 0.001) . The proportion of patients with severe BP was significantly higher in the DPP4i-BP group (16 cases, 50%) than in the general BP group (25 cases, 29.76%; Z= 2.63, P= 0.008) . There was no significant difference in the positivity rate of anti-BP180 antibody between the two groups ( χ2= 0.03, P= 0.870) . However, the level of anti-BP180 antibody was significantly higher in the DPP4i-BP group than in the general BP group before and after treatment ( P= 0.015, < 0.001, respectively) , and the decrease in the level of anti-BP180 antibody was significantly less in the DPP4i-BP group than in the general BP group after treatment ( t= 5.11, P < 0.001) . There was no significant difference in the average effective dose of glucocorticoids required to control the disease between the two groups ( t= 1.00, P= 0.322) . However, the DPP4i-BP group showed a significant increase in the average time required to control the disease and in the proportion of patients requiring combined treatment with immunosuppressants or other drugs compared with the general BP group ( t= 6.72, 10.05, P < 0.001,= 0.002, respectively) . Within 6 months after the start of systemic treatment, the recurrence rate was significantly higher in the general BP group (17 cases, 27.86%) than in the DPP4i-BP group (2 cases, 7.69%; χ2= 4.35, P= 0.037) ; at 6 months, the average dose of glucocorticoids was also significantly higher in the general BP group than in the DPP4i-BP group ( t= 7.04, P < 0.001) . Conclusions:Among the DPP4i hypoglycemic drugs, vildagliptin was the most common drug administrated by patients before the onset of BP, with the shortest interval from the administration to the onset of BP. DPP4i-BP may be difficult to control at the early stage, but the prognosis is good.

In recent years, the genome editing technologies based on the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) have developed rapidly. The system can use homologous directed recombination (HDR) to achieve precise editing that it medicated, but the efficiency is extremely low, which limits its application in agriculture and biomedical fields. As an emerging genome editing technology, the CRISPR/Cas-mediated DNA base editing technologies can achieve targeted mutations of bases without generating double-strand breaks, and has higher editing efficiency and specificity compared with CRISPR/Cas-mediated HDR editing. At present, cytidine base editors (CBEs) that can mutate C to T, adenine base editors (ABEs) that can mutate A to G, and prime editors (PEs) that enable arbitrary base conversion and precise insertion and deletion of small fragments, have been developed. In addition, glycosylase base editors (GBEs) capable of transitioning from C to G and double base editors capable of editing both A and C simultaneously, have been developed. This review summarizes the development, advances, advantages and limitations of several DNA base editors. The successful applications of DNA base editing technology in biomedicine and agriculture, together with the prospects for further optimization and selection of DNA base editors, are discussed.
Agriculture ; CRISPR-Cas Systems/genetics* ; DNA/genetics* ; Gene Editing ; Technology

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.
CRISPR-Cas Systems ; DNA Ligase ATP ; genetics ; Gene Expression Regulation ; genetics ; Gene Knockdown Techniques ; HEK293 Cells ; Humans ; Ku Autoantigen ; genetics

Objective To identify the pathogenesis gene mutation of a pedigree with Cockayne syndrome.Methods The peripheral blood samples of the patient and his family members were collected and the genomic DNA was then extracted.Whole exome sequencing(WES)was performed for proband′s DNA.The disease-causing mutations were identified by bioinformatics analysis and pedigree analysis. Meanwhile,the mutations were confirmed by Sanger sequencing.Results Two novel mutations in ERCC8 gene,including c.400-2A >G and c.394_398delATGTA(p.L132fs)were identified in proband.The splicing mutation originated from his father and changed the splice acceptor site AG to GG, thus possibly caused alternative splicing.The c.394_398delATGTA(p.L132fs)frameshifting mutation was inherited from his mother.The proband′s sister also carried the same compound heterozygous mutation and had the same phenotype as proband.Conclusion The pathogenesis ERCC8 gene mutation of this pedigree with Cockayne syndrome was identified by using whole exome sequencing.

Objective:To explore the regional brain activities in healthy adolescent subjects after sleep deprivation (SD) using amplitude of low-frequency fluctuation (ALFF) method.Methods:Total of 16 healthy adolescent subjects (8 males,8 females;aged 13-20 years) were recruited in the community and the campus through the internet and posters.Each of the 16 healthy adolescent subject underwent the attention network test and magnetic resonance imaging (MRI) session twice:once was after rested wakefulness (RW condition),and the other was after SD condition.Amplitude of low frequency fluctuation (ALFF) method was used to assess the local brain features.The mean ALFF signal values of the different brain areas were performed to investigate their relationships with the accuracy rate,reaction time and lapse rate in the attention network test,and were analyzed with a receiver operating characteristic (ROC) curve to investigate their sensitivities and specificities to distinguish the SD condition from the RW condition.Results:Subjects showed a lower response accuracy rate [(83 ± 12) ％ vs.(97 ± 4) ％,P ＜ 0.05],a longer response time [(832 ± 134) ms vs.(715 ± 97) ms,P ＜ 0.05] and a higher lapse rate [(15 ± 11)％ vs.(2.4 ±7.3)％,P ＜0.05] under SD condition than under RW condition.They showed higher ALFF area in the right cuneus (BA 17,BA 18),and lower ALFF areas in the right lentiform nucleus,right claustrum,left dorsolateral prefrontal cortex (BA 46) and left inferior parietal cortex (BA 39) under SD condition than under RW condition.Under SD condition,the mean ALFF signal value of the right claustrum showed a significant positive correlation with the accuracy rate (r =0.69,P ＜0.05),and a negative correlation with the lapse rate (r =-0.71,P ＜0.05).The mean ALFF signal value of the dorsolateral prefrontal cortex showed a significant positive correlation with the reaction time (r =0.68,P ＜ 0.05).The values of area under the curve of the right cuneus,right lentiform nucleus,right claustrum,left dorsolateral prefrontal cortex and left inferior parietal cortice were 0.9,0.8,0.9,0.8 and 0.9,respectively.These different ALFF areas also showed high degree of sensitivities and specificities.Conclusion:Sleep deprivation leads to the dysfunction in the default mode network,anticorrelatedtask-positive network,and advanced cognitive function brain areas,and the functional compensation in the visual network.

Objective To study related factors of the duration of untreated psychosis (DUP) in Tibetan patients with schizophrenia in Qinghai Province. Methods The related factors on DUP were investigated in totally 188 Tibetan pa?tients with schizophrenia using questionnaires of mental health services and symptom onset for schizophrenia. All the Ti?betan patients were provided with the National Continuing Management and Intervention Program for Psychoses (686 Pro?gram). Results The median (low quartile, upper quartile) of DUP in Tibetan patients with schizophrenia was 375 days (4 days, 1661 days). The patients were divided into short DUP group (DUP≤375days, 90 patients) and long DUP group (DUP>375 days, 98 patients). There were significant differences in mode of onset,marital status, educational level, family type, place of residence between short DUP group and long DUP group (P<0.05). Logistic regression analysis found that lack of family structure (OR=2.340, 95%CI:1.130~4.847, P=0.022), chronic onset (OR=2.136, 95%CI:1.172~3.891, P=0.013) and living in pastoral areas (OR=2.239, 95%CI:1.097~4.571, P=0.027) were risk factors of DUP. Conclusion Ti?betan patients with schizophrenia have a longer DUP and related risk factories of DUP are lack of family structure, chron?ic onset and living in pastoral areas.