Objective:To investigate the molecular mechanism of the 5-methylcytosine(m5C)reader Y-box binding protein 1(YBX1)in participating in the development and progression of colorectal cancer(CRC)by regulating the stability of the ferroptosis inhibitor membrane-spanning 4-domains subfamily A 15(MS4A15).Methods:Bioinformatics databases were used to investigate the mRNA and protein expression levels of YBX1 in CRC.RT-qPCR was used to measure the expression level of YBX1 in CRC cells.LC-MS was used to measure the level of m5C modification in CRC cells and nor-mal colorectal mucosal cells.CCK-8 assay was used to observe the effect of YBX1 on the proliferation of CRC cells,Transwell assay was used to observe its effect on the migration ability of CRC cells,and flow cytometry was sued to observe its effect on the apoptosis of CRC cells.Bioinformatics methods were used to identify the ferrop-tosis inhibitors that can interact with YBX1 and potential m5C modification sites.GEPIA2 was used to analyze the association between the expression of YBX1 and MS4A15.The expression of YBX1 was inhibited,and then the mRNA expression level and m5C modifica-tion level of MS4A15 were analyzed.The catRAPID database was used to find the binding sites between YBX1 protein and MS4A15 mRNA.CRC cells were treated with actinomycin D after inhibition of YBX1 expression,and RT-qPCR was used to measure the stabil-ity of MS4A15 mRNA.The expression of MS4A15 was inhibited,and then the proliferative activity,migration ability,and apoptosis rate of cells were measured,as well as the expression levels of the key indicators for ferroptosis,including MDA,ROS,and Fe2+.Results:High mRNA and protein expression levels of YBX1 were observed in CRC,and YBX1 was highly expressed in CRC cells.The m5C modification level of CRC cells was significantly higher than that of normal colorectal mucosal cells.YBX1 could promote the prolifera-tion and migration of CRC cells and inhibit the apoptosis of CRC cells.The bioinformatics analysis showed that YBX1 was positively correlated with the expression level of the ferroptosis inhibitor MS4A15,and there were multiple m5C modification sites on MS4A15.Inhibition of YBX1 expression could reduce the mRNA expression level and m5C modification level of MS4A15 and the stability of MS4A15 mRNA.There were significant reductions in the proliferative activity and migration ability of CRC cells and a significant in-crease in the apoptosis rate of CRC cells after inhibition of MS4A15 expression,with significant increases in the content of MDA,ROS,and Fe2+.Conclusion:These results show that YBX1 promotes the development and progression of CRC by stabilizing MS4A15 via m5C modification,which provides a promising targeted therapeutic strategy for CRC patients.

Objective To analyze the computed tomography(CT)imaging features of urachal carcinoma and evaluate its diagnostic value.Methods The clinical data of 20 patients with urachal carcinoma confirmed by surgery and pathology,who were admitted to The First Affiliated Hospital of Naval Medical University from Dec.2012 to Dec.2022,were collected.Seventeen of the 20 patients underwent enhanced CT urography and 3 underwent pelvic CT plain scan+enhanced scan.After scanning,multiplanar reconstruction was performed on the post-processing workstation.The general data,clinical symptoms,CT imaging findings,pathological data,and prognosis of the patients were analyzed and summarized.Results The patients included 16 males and 4 females,aged 27 to 75 years old,with a median age of 61.50(41.50,71.25)years old.The tumors were all located in the anterior wall of the bladder,along the urachus,with a maximum diameter of 1.72-5.55 cm and a median maximum diameter of 3.34(2.48,3.71)cm.Fourteen cases had cystic-solid lesions and 6 had solid lesions.In the cystic-solid lesions,9 cases showed the"upper cystic and lower solid"sign on the sagittal plane.Calcification was noted in 17 cases.After enhanced scanning,18 cases showed progressive enhancement,and 2 cases showed"fast in and fast out"enhancement.Tumor invasion extended beyond the urachus and/or bladder muscle layer in 19 cases.At the end of follow-up,3 cases had recurrence,2 had metastasis,5 had no recurrence after surgery,3 died,and 7 were lost to follow-up.Conclusion Urachal carcinoma has certain characteristic manifestations on CT imaging.Reconstructing the sagittal plane with enhanced CT scanning and multiplanner reformation can help preoperative diagnosis and prognostic evaluation of urachal carcinoma.

Objective:To explore the strategy of prostate biopsy in patients with prostate specific antigen(PSA)gray zone based on prostate imaging reporting and data system (PI-RADS).Methods:The clinical data of 427 patients who underwent transperineal prostate biopsy in the First Affiliated Hospital of Nanjing Medical University from January 2020 to December 2022 were retrospectively analyzed. The median age was 66 (61, 72) years old. The median PSA was 6.62 (5.46, 8.19) ng/ml. The median PSA density (PSAD) was 0.15 (0.11, 0.21) ng/ml 2. The median prostate volume (PV) was 43.68 (31.12, 56.82) ml. PSA velocity (PSAV) data were available in 65 patients with negative MRI examination(PI-RADS <3), and the median PSAV was 1.40 (0.69, 2.89) ng/(ml· year). Among the patients with positive MRI(PI-RADS≥3), there were 174 patients with only 1 lesion and 83 patients with ≥2 lesions. A total of 170 patients with negative MRI underwent systematic biopsy, and 257 patients with positive MRI underwent systematic combined targeted biopsy. The PI-RADS score, regions of interest(ROI), PSAD, f/tPSA and PSAV were analyzed to explore the biopsy strategy for patients with PSA gray area based on bpMRI imaging. Results:Of the 427 patients included in the study, 194 were positive and 233 were negative. Among the patients with positive biopsy pathology, 140 cases were clinically significant prostate cancer (CsPCa). Among the MRI-negative patients, there were 33 cases with PSAV ≥1.4 ng/(ml·year), and 10 cases of prostate cancer and 6 cases of CsPCa were detected by systematic biopsy.In 32 cases with PSAV <1.4 ng/(ml·year), 3 cases of prostate cancer and 0 case of CsPCa were detected by systematic biopsy. The sensitivity of systematic biopsy for the diagnosis of prostate cancer and CsPCa in patients with PSAV≥1.4 ng/(ml·year) were 76.9% (10/13) and 100.0% (6/6) respectively, the specificity were 55.8% (29/52) and 54.2% (32/59) respectively, the negative predictive value were 90.6% (29/32) and 100.0% (32/32) respectively, and the positive predictive value were 30.3% (10/33) and 18.2% (6/33) respectively. In MRI-positive patients with PI-RADS 3, the prostate cancer detection rates of targeted biopsy combined with systematic biopsy, systematic biopsy and targeted biopsy were 41.7% (45/108), 32.4% (35/108) and 35.2% (38/108), respectively ( P=0.349). The detection rates of CsPCa were 27.8% (30/108), 21.3% (23/108) and 25.0% (27/108), respectively ( P=0.541). In patients with PI-RADS 4-5 and PSAD > 0.15 ng/ml 2, the detection rates of CsPCa in targeted biopsy combined with systematic biopsy, systematic biopsy and targeted biopsy were 67.8% (61/90), 58.9% (53/90) and 67.8% (61/90), respectively ( P=0.354). Conclusions:For MRI-negative patients, all CsPCa could be detected by perineal systematic biopsy when PSAV ≥1.4 ng/(ml·year), and active observation could be performed when PSAV <1.4 ng/(ml·year). For MRI-positive patients, targeted combined systemic biopsy was required when PI-RADS score was 3, and targeted biopsy only could be performed when PI-RADS score ≥4 and PSAD >0.15 ng/ml 2, otherwise targeted combined systemic biopsy was required.

Objective:To analyze the clinical features and magnetic resonance imaging of non-malignant patients assigned to Prostate Imaging Reporting And Data System (PI-RADS) 5 score.Methods:We performed a retrospective review of 289 patients who underwent magnetic resonance ultrasound targeted combined system biopsy with PI-RADS 5 lesions in the First Affiliated Hospital of Nanjing Medical University between May 2019 and July 2021. The median age 72 (66, 77)years, median body mass index 24.4(22.3, 27.1)kg/m 2, median prostate volume (PV) 37.39(29.39, 48.86) ml, median PSA 22.24(10.91, 62.69) ng/ml, and median PSAD 0.53(0.30, 1.52)ng/ml 2 were recorded. According to the biopsy pathological results, all patients were divided into benign lesion group and prostate cancer group. PSA, PSAD, PV, and apparent diffusion coefficient (ADC) values were compared, and magnetic resonance imaging and clinical characteristics of patients with biopsy benign lesions were analyzed. Results:There were 11 cases (3.8%) with benign lesion and 278 cases (96.2%) with prostate cancer. The characters of 11 negative biopsy cases were displayed as follows: median age 69(66, 79)years, median body mass index 22.0(21.0, 25.5)kg/m 2, median PV 62.90(38.48, 71.96)ml, median PSA 5.55(2.99, 20.52)ng/ml, median PSAD 0.16(0.07, 0.24) ng/ml 2, median ADC 714.47(701.91, 801.26)×10 -6 mm 2/s, abnormal digital rectal and amination in 5 cases, smoking in 7 cases, and alcohol consumption in 4 cases. The median PV [62.90(38.48, 71.96) vs. 37.21(29.22, 47.82)ml, P<0.01], the PSA value [5.55(2.99, 20.52) vs. 23.53(11.14, 65.98)ng/ml, P<0.01], and the PSAD value [0.16(0.07, 0.24) vs. 0.58(0.31, 1.57)ng/ml 2, P<0.01] were significantly different between benign condition group and prostate carcinoma group. Benign condition group included 5 chronic prostatitis, 2 acute prostatitis (1 with focal adenocarcinoma), 2 granulomatous inflammation, and 2 tuberculous granulomatous inflammation. In 7 benign cases, PSA was less than 10 ng/ml, combined with frequent urination, urgency of urination and incontinence were founded. In 8 benign cases, the area of lesion was more than 50% of the total prostate area in the axial position and the imaging of magnetic resonance were diffused, with regular shape and uniform signal. The imaging of symmetrical distribution was in 6 cases. Conclusions:The benign condition with PI-RADS 5 lesions included chronic prostatitis, acute prostatitis, granulomatous inflammation and tuberculous granulomatous inflammation, among which prostatitis was the most common cause. The PSA value were less than 10 ng/ml in most benign cases, with symptoms such as frequent urination, urgency of urination and incontinence. The imaging of magnetic resonance were diffused, symmetrically distributed, with regular shape and uniform signal.

Objective:To evaluate the effects of remimazolam on gastrointestinal motor function in the patients undergoing gastrointestinal endoscopy.Methods:A total of 262 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-64 yr, with body mass index of 18-28 kg/m 2, scheduled for elective gastrointestinal endoscopy from May 2022 to August 2022, were divided into 2 groups ( n=131 each) using a random number table method: remimazolam group (group R) and propofol group (group P). The patients in group R received intravenous remimazolam 0.20-0.25 mg/kg, and patients in group P received intravenous propofol 1.5-2.0 mg/kg. The gastrointestinal endoscopy was performed when the patients′ Modified Observer′s Assessment of Alertness/Sedation scores ≤3. During fasting before gastrointestinal preparation, before gastrointestinal endoscopy and while leaving the post-anesthesia care unit (PACU), the concentrations of serum motilin and gastrin were measured by enzyme-linked immunosorbent assay, the intestinal peristalsis rating assessed by the endoscopist during the examination was recorded, the occurrence of hypotension and hypoxemia during the examination and occurrence of abdominal distension, abdominal pain, and nausea and vomiting during stay in PACU were recorded. Results:Compared with group P, the intestinal peristalsis rating was significantly increased, the serum motilin and gastrin concentrations were increased while leaving PACU, the incidence of hypotension and hypoxemia was decreased during the examination, and the incidence of abdominal distention was decreased during stay in PACU in group R ( P<0.05). Conclusions:Remimazolam has a milder inhibitory effect on secretion of gastrointestinal hormones than propofol in the patients undergoing gastrointestinal endoscopy and is helpful for the recovery of gastrointestinal motility.

Objective:To evaluate the role of heat shock transcription factor 1 (HSF1) in the endogenous protective mechanism underlying mechanical ventilator-induced lung injury (VILI) in mice and the relationship with high mobility group box 1 (HMGB1).Methods:Forty SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) by the random number table method: control group (group C), VILI group (group VILI), negative control siRNA + VILI group (group NV) and HSF1 siRNA + VILI group (group siRNA). At 48 h before mechanical ventilation, negative control siRNA 5 nmol and HSF1 siRNA 5 nmol were intratracheally injected in NV and siRNA groups respectively, and the solution was diluted to 50 μl with the sterile phosphate buffer in both groups. Group C kept spontaneous breathing for 4 h, and the rest animals were mechanically ventilated (tidal volume 35 ml/kg, respiratory rate 75 breaths/min, inspiratory/expiratory ratio 1∶2, fraction of inspired oxygen 21%) for 4 h. Blood samples from the femoral artery were collected for arterial blood gas analysis immediately after endotracheal intubation and at 4 h of ventilation, and PaO 2 was recorded. Then the mice were sacrificed under deep anesthesia to collect lung tissues and bronchoalveolar lavage fluid (BALF). The concentrations of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and HMGB1 in BALF were determined by enzyme-linked immunosorbent assay. The pathological results were observed by hematoxylin-eosin staining, and lung injury was assessed and scored. The wet/dry (W/D) weight ratio of lung tissues was calculated. The expression of HMGB1 and HSF1 mRNA in lung tissues (by quantitative real-time polymerase chain reaction) and expression of HMGB1 and HSF1 protein in lung tissues (by Western blot) were determined. Results:Compared with group C, PaO 2 was significantly decreased at 4 h of ventilation, the concentrations of TNF-α, IL-1β and HMGB1 in BALF, W/D ratio and lung injury score were increased, and the expression of HMGB1 protein and mRNA in lung tissues was up-regulated in group VILI, group NV and group siRNA ( P<0.05 or 0.01). Compared with group VILI and group NV, PaO 2 was significantly decreased at 4 h of ventilation, the concentrations of TNF-α, IL-1β and HMGB1 in BALF, W/D ratio and lung injury score were increased, and the expression of HMGB1 protein and mRNA in lung tissues was up-regulated, and the expression of HSF1 protein and mRNA was down-regulated in group siRNA ( P<0.05 or 0.01). There was no significant difference in the parameters mentioned above between group VILI and group NV ( P>0.05). Conclusions:HSF1 is involved in the endogenous protective mechanism underlying VILI in mice, which may be related to the down-regulation of HMGB1 expression and attenuation of inflammatory responses in lung tissues.

Objective To explore the application value of automatic cytomorphological analyzer in the morphological analysis of white blood cells(WBC)in peripheral blood.Methods A total of 306 venous blood samples from inpatients and outpatients were randomly selected and prepared with automatic cytomorphological analyzer for WBC pre-classification.The differences between automatic cytomorphological analyzer counting,automatic blood cell analyzer counting and manual counting were compared,and the correlation between automatic cytomorphological analyzer and manual counting method was analyzed.Results Compared with the other two methods,the automatic cytomorphologi-cal analyzer was able to detect more types of WBC,especially abnormal cells.There were no signifi-cant differences between automatic cytomorphological analyzer and manual counting method for 6 ma-ture WBC types(band neutrophils,segmented neutrophils,lymphocytes,monocytes,eosinophils,and basophils),immature cells at different stages and atypical lymphocyte counts(P＞0.05).Re-sults of the 6 mature WBC types counted by the automatic cytomorphological analyzer and manual counting had favorable correlations(r＞0.8).Conclusion The automatic cytomorphological analyzer can classify more types of WBC,provide WBC counting results that are highly consistent with manual microscopy,and the counting results of the two methods have a good correlation.

Objective To explore the application value of automatic cytomorphological analyzer in the morphological analysis of white blood cells(WBC)in peripheral blood.Methods A total of 306 venous blood samples from inpatients and outpatients were randomly selected and prepared with automatic cytomorphological analyzer for WBC pre-classification.The differences between automatic cytomorphological analyzer counting,automatic blood cell analyzer counting and manual counting were compared,and the correlation between automatic cytomorphological analyzer and manual counting method was analyzed.Results Compared with the other two methods,the automatic cytomorphologi-cal analyzer was able to detect more types of WBC,especially abnormal cells.There were no signifi-cant differences between automatic cytomorphological analyzer and manual counting method for 6 ma-ture WBC types(band neutrophils,segmented neutrophils,lymphocytes,monocytes,eosinophils,and basophils),immature cells at different stages and atypical lymphocyte counts(P＞0.05).Re-sults of the 6 mature WBC types counted by the automatic cytomorphological analyzer and manual counting had favorable correlations(r＞0.8).Conclusion The automatic cytomorphological analyzer can classify more types of WBC,provide WBC counting results that are highly consistent with manual microscopy,and the counting results of the two methods have a good correlation.

Objective:To evaluate the role of inositol-requiring kinase 1α-X-box binding protein 1 (IRE1α/XBP1) signaling pathway in endoplasmic reticulumin in endotoxin-induced acute lung injury (ALI) in mice and the relationship with NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasomes.Methods:Thirty-six SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 25-30 g, were divided into 3 groups ( n=12 each) according to the random number table method: control group (group C), endotoxin-induced ALI group (group ALI) and endotoxin-induced ALI+ STF-083010 group (group ST). The ALI model was established by inhaling nebulized lipopolysaccharide (LPS) 3 mg/ml for 30 min in ALI and ST groups, while the equal volume of nebulized normal saline was given in group C. IRE1α/XBP1 signaling pathway inhibitor STF-083010 50 mg/kg was intraperitoneally injected at 1 h before inhaling LPS in group ST, while the remaining two groups received the equal volume of normal saline intraperitoneally. Mice were sacrificed at 24 h after inhaling nebulized LPS or normal saline, bronchoalveolar lavage fluid (BALF) were collected and lung tissues were removed for microscopic examination of the pathological changes (by HE staining) which were scored and for determination of wet/dry lung weight ratio (W/D ratio), concentrations of interleukin-1beta (IL-lβ) and IL-18 in BALF (by enzyme-linked immunosorbent assay) and expression of phosphorylated IRE1α (p-IRE1α), XBP1s, NLRP3, ASC and caspase-1 in lung tissues (by Western blot). Results:Compared with group C, the W/D ratio, lung injury score and concentrations of IL-1β and IL-18 in BALF were significantly increased, and the expression of p-IRE1α, XBP1s, NLRP3, ASC and caspase-1 in lung tissues was up-regulated in group ALI and group ST ( P<0.001). Compared with group ALI, the W/D ratio, lung injury score and concentrations of IL-1β and IL-18 in BALF were significantly decreased, and the expression of p-IRE1α, XBP1s, NLRP3, ASC and caspase-1 protein in lung tissues was down-regulated in group ST ( P<0.001). Conclusions:IRE1α/XBP1 signaling pathway is involved in LPS-induced ALI in mice, and the mechanism is related to activation of NLRP3 inflammasomes.

Objective:To explore the influencing factors of clinically significant prostate cancer (CsPCa) in patients with PI-RADS score 3.Methods:The data of 133 consecutive patients with the PI-RADS score 3 from January 2019 to December 2020 were retrospectively analyzed. All patients underwent 4-needle transperineal targeted biopsy and 12-needle systematic prostate biopsy (SB). The overall age was 66 (60-72) years, and the overall PSA value was 8.22 (5.95-11.41) ng/ml. All patients underwent multiparametric magnetic resonance imaging (mpMRI), and PI-RADS v2.0 score was 3. Patients were divided into two mutually exclusive groups: non CsPCa group and CsPCa group. The differences of lesion location, laterality, focality and sequence parameters of mpMRI between the two groups were compared, and multivariate binary logistic regression was used to analyze the independent predictors of PI-RADS score 3 in patients with CsPCa.Results:Biopsy results showed 57 cases of prostate cancer, including 41 cases of CsPCa, and 76 cases of non-prostate cancer. The detection rate of prostate cancer was 46.62 %(57/133), and the detection rate of CsPCa was 30.83 %(41/133). There were 41 cases in CsPCa group and 92 cases in non CsPCa group. There was no significant difference between CsPCa group and non CsPCa group in age [66 (58-70) years vs. 66 (60-72) years], body mass index [24.22 (21.82-25.71) kg/m 2 vs. 23.71 (21.99-26.12) kg/m 2], PSA [9.39 (6.35-12.55) ng/ml vs. 7.67 (5.83-10.51) ng/ml], abnormal rate of rectal digital examination [21.95% (9/41) vs. 9.78% (9/92)] (all P > 0.05). There was significant difference in PSAD [0.40 (0.16-0.65) ng/ml 2 vs. 0.17 (0.12-0.24) ng/ml 2] ( P<0.05). In MRI, PI-RADS=3 lesions were mainly located in the transitional zone [46.62 %(62/133)]. In CsPCa group, MRI lesions were located in peripheral zone in 16 cases, transitional zone in 19 cases, and both areas in 6 cases. There were 16 cases on the right, 15 cases on the left and 10 cases on both sides. The lesions were diffused in 19 cases and localized in 22 cases. In the non CsPCa group, 41 lesions were located in the peripheral zone, 43 in the transitional zone, and 8 in both areas. There were 26 cases on the right, 35 cases on the left and 31 cases on both sides. The lesions were diffuse in 56 cases and localized in 36 cases. There was no significant difference in lesion location, side and diffusion degree between the two groups ( P> 0.05). Compared with the non CsPCa group, the positive rate of all MRI sequences in CsPCa group was higher (82.93% vs. 40.22%, P < 0.001), the positive rate of T2 weighted imaging (T2WI) was higher (92.68% vs. 75.00%, P = 0.018), the positive rate of diffusion weighted imaging (DWI) was higher (90.24% vs. 56.52%, P < 0.001), the maximum diameter was larger[(0.67(0.30-1.19)mm vs. 0.48(0.20-0.62)mm, P < 0.001], and the apparent diffusion coefficient (ADC) was lower[0.70(0.61-0.87) vs. 1.10(0.86-1.50), P < 0.001]. Concurrently, PSAD and lesion ADC were important predictors of CsPCa in logistic regression model [mean 10 fold cross validation AUC: 0.78(95% CI 0.65-0.88)]. Conclusions:Most of the MRI lesions in patients with PI-RADS 3 were located in the transitional zone, and the MRI lesions in CsPCa were more obvious and diffusion limited. PSAD and ADC values are independent predictors for the diagnosis of CsPCa in patients with PI-RADS score 3, and the log 2PSAD-ADC prediction model is helpful to find CsPCa from patients with PI-RADS score 3 and protect patients from unnecessary biopsy.