Objective:To explore the molecular mechanism by which interferon regulatory factor 1 (IRF1) affects hepatic ischemia-reperfusion injury (HIRI) by regulating the polarization of Kupffer cells.Methods:Twelve male healthy C57BL/6 wild-type mice weighing 20-25 g and aged 6-8 weeks were divided into a sham operation group ( n=6) and a HIRI group ( n=6); Twelve male healthy C57BL/6 IRF1 gene knockout (IRF1 -/-) mice weighing 20-25 g and aged 6-8 weeks were divided into a sham operation IRF1 -/- group ( n=6) and a HIRI IRF1 -/- group ( n=6). The levels of serum alanine transaminase (ALT) and aspartate transaminase (AST) in mice were measured, and hematoxylin-eosin (HE) staining of liver tissues was performed for Suzuki scoring to evaluate liver injury. Fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to evaluate the mRNA levels of IRF1 and tumor necrosis factor α (TNFα) in liver tissues. Flow cytometry and qRT-PCR were used to detect the proportion and functional changes of M1/M2-type Kupffer cells in liver tissues. IRF1 was overexpressed or knocked down in the mononuclear macrophage cell line ANA1, and a co-culture and hypoxia-reoxygenation system with the hepatocyte cell line AML12 was established. Flow cytometry was used to detect the apoptosis of AML12 cells. Results:At 12 hours after hepatic ischemia-reperfusion in wild-type mice, the liver tissue injury was the most severe. Compared with the sham operation group, the levels of serum ALT [(8 073±83) U/L vs. (81±19) U/L, q=13.59] and AST [(11 170±2 890) U/L vs. (412±210) U/L, q=13.77] in the HIRI group were significantly higher, and the differences were statistically significant (both P<0.001). The Suzuki score reached 5-6 points. At 12 hours after hepatic ischemia-reperfusion in IRF1 gene knockout mice, the liver tissue injury was not obvious. There were no significant differences in the levels of serum ALT [668 (514, 2 344) U/L vs. 254 (147, 285) U/L, q=2.52, P=0.348] and AST [1 936 (1 262, 2 003) U/L vs. 628 (423, 759) U/L, q=1.22, P=0.824] between the HIRI IRF1 -/- group and the sham operation IRF1 -/- group. Compared with the HIRI group, the ratio of M1/M2-type Kupffer cells in the liver of the HIRI IRF1 -/- group decreased [(0.958±0.090) vs. (2.788±0.258), q=2.06, P<0.0001], and the mRNA expression of TNFα decreased [(4.363±0.393) vs. (12.900±5.504), q=5.59, P=0.018], and the differences between the two groups were statistically significant. In the co-culture and hypoxia-reoxygenation experiment using ANA1 cells overexpressing IRF1 and AML12 cells, the proportion of AML12 hepatocytes in late apoptosis was higher than that in the control group [(14.05±4.25) vs. (3.15±1.16), t=2.85, P=0.047], and the difference was statistically significant. In contrast, when the expression of IRF1 was knocked down, the proportion of apoptotic AML12 cells decreased [(9.26±3.04) vs. (13.36±4.64), t=2.15, P=0.098], but the difference was not statistically significant. Conclusion:The IRF1 protein can regulate the polarization of Kupffer cells into M1-type macrophages, promote the inflammatory injury of the liver tissue after ischemia-reperfusion, and increase the apoptosis of hepatocytes.

[Objective] To analyze the loss rate of platelet donors and evaluate the strategies for establishing a platelet donor bank. [Methods] A total of 1 443 donors who joined the HLA and HPA gene donor bank for platelets in Henan Province from 2018 to 2020 were included in this study. Data on the total number of apheresis platelet donations, annual donation frequency, age at enrollment, donation habits (including the number of platelets donated per session and whether they had previously donated whole blood), and enrollment location were collected from the platelet donor information management system. Donor loss was determined based on the date of their last donation. The loss rates of different groups under various conditions were compared to assess the enrollment strategies. [Results] By the time the platelet bank was officially operational in 2022, 421 donors had been lost, resulting in an loss rate of 29% (421/1 443). By the end of 2023, the overall cumulative loss rate reached 52% (746/1 443). The loss rate was lower than the overall level in groups meeting any of the following conditions: total apheresis platelet donations exceeding 50, annual donation frequency of 10 or more, age at enrollment of 40 years or older, donation of more than a single therapeutic dose per session, or a history of whole blood donation two or more times. Additionally, loss rates varied across different enrollment locations, with higher enrollment numbers generally associated with higher loss rates. [Conclusion] Through a comprehensive analysis of donor loss, our center has adjusted its strategies for establishing the donor pool. These findings also provide valuable insights for other blood collection and supply institutions in building platelet donor banks.

Objective To evaluate the feasibility of fluoroscopy-guided anterograde forceps biopsy for diagnosing uretero-ileal neobladder anastomotic stenosis.Methods Twenty-two patients who underwent Bricker surgery presented with hydronephrosis were selected.Using a catheter-wire technique,percutaneous nephrostomy tract was used to access the uretero-ileal neobladder stric-ture and establish a sheath biopsy channel.Forceps biopsy was performed at the uretero-ileal neobladder anastomotic site,followed by placement of a 10.2F ureteral stent.The technical success rate,complications,biopsy sensitivity,specificity,accuracy were recorded,and preoperative versus postoperative white blood cell counts,creatinine,and urea nitrogen levels were compared.Results The tech-nical success rate was 100％(22/22),with no serious complications,such as ureteral perforation and major bleeding.The accuracy,sensitivity and specificity of the biopsy were 95.45％(21/22),85.71％(6/7)and 100％(15/15),respectively.Preoperative and post-operative white blood cell counts were(9.17±2.16)× 1012/L vs(6.03±1.51)×1012/L,creatinine levels were(219.95±78.47)U/mL vs(78.91±17.23)U/mL,and urea nitrogen levels were(19.85±5.27)U/mL vs(5.95±1.60)U/mL.All three parameters showed statistically significant differences(P＜0.05).Conclusion The fluoroscopy-guided anterograde forceps biopsy for diagnosing uretero-ileal neobladder anastomotic stenosis is safe and feasible.

Objective To evaluate the feasibility of fluoroscopy-guided anterograde forceps biopsy for diagnosing uretero-ileal neobladder anastomotic stenosis.Methods Twenty-two patients who underwent Bricker surgery presented with hydronephrosis were selected.Using a catheter-wire technique,percutaneous nephrostomy tract was used to access the uretero-ileal neobladder stric-ture and establish a sheath biopsy channel.Forceps biopsy was performed at the uretero-ileal neobladder anastomotic site,followed by placement of a 10.2F ureteral stent.The technical success rate,complications,biopsy sensitivity,specificity,accuracy were recorded,and preoperative versus postoperative white blood cell counts,creatinine,and urea nitrogen levels were compared.Results The tech-nical success rate was 100％(22/22),with no serious complications,such as ureteral perforation and major bleeding.The accuracy,sensitivity and specificity of the biopsy were 95.45％(21/22),85.71％(6/7)and 100％(15/15),respectively.Preoperative and post-operative white blood cell counts were(9.17±2.16)× 1012/L vs(6.03±1.51)×1012/L,creatinine levels were(219.95±78.47)U/mL vs(78.91±17.23)U/mL,and urea nitrogen levels were(19.85±5.27)U/mL vs(5.95±1.60)U/mL.All three parameters showed statistically significant differences(P＜0.05).Conclusion The fluoroscopy-guided anterograde forceps biopsy for diagnosing uretero-ileal neobladder anastomotic stenosis is safe and feasible.

Objective:To explore the molecular mechanism by which interferon regulatory factor 1 (IRF1) affects hepatic ischemia-reperfusion injury (HIRI) by regulating the polarization of Kupffer cells.Methods:Twelve male healthy C57BL/6 wild-type mice weighing 20-25 g and aged 6-8 weeks were divided into a sham operation group ( n=6) and a HIRI group ( n=6); Twelve male healthy C57BL/6 IRF1 gene knockout (IRF1 -/-) mice weighing 20-25 g and aged 6-8 weeks were divided into a sham operation IRF1 -/- group ( n=6) and a HIRI IRF1 -/- group ( n=6). The levels of serum alanine transaminase (ALT) and aspartate transaminase (AST) in mice were measured, and hematoxylin-eosin (HE) staining of liver tissues was performed for Suzuki scoring to evaluate liver injury. Fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to evaluate the mRNA levels of IRF1 and tumor necrosis factor α (TNFα) in liver tissues. Flow cytometry and qRT-PCR were used to detect the proportion and functional changes of M1/M2-type Kupffer cells in liver tissues. IRF1 was overexpressed or knocked down in the mononuclear macrophage cell line ANA1, and a co-culture and hypoxia-reoxygenation system with the hepatocyte cell line AML12 was established. Flow cytometry was used to detect the apoptosis of AML12 cells. Results:At 12 hours after hepatic ischemia-reperfusion in wild-type mice, the liver tissue injury was the most severe. Compared with the sham operation group, the levels of serum ALT [(8 073±83) U/L vs. (81±19) U/L, q=13.59] and AST [(11 170±2 890) U/L vs. (412±210) U/L, q=13.77] in the HIRI group were significantly higher, and the differences were statistically significant (both P<0.001). The Suzuki score reached 5-6 points. At 12 hours after hepatic ischemia-reperfusion in IRF1 gene knockout mice, the liver tissue injury was not obvious. There were no significant differences in the levels of serum ALT [668 (514, 2 344) U/L vs. 254 (147, 285) U/L, q=2.52, P=0.348] and AST [1 936 (1 262, 2 003) U/L vs. 628 (423, 759) U/L, q=1.22, P=0.824] between the HIRI IRF1 -/- group and the sham operation IRF1 -/- group. Compared with the HIRI group, the ratio of M1/M2-type Kupffer cells in the liver of the HIRI IRF1 -/- group decreased [(0.958±0.090) vs. (2.788±0.258), q=2.06, P<0.0001], and the mRNA expression of TNFα decreased [(4.363±0.393) vs. (12.900±5.504), q=5.59, P=0.018], and the differences between the two groups were statistically significant. In the co-culture and hypoxia-reoxygenation experiment using ANA1 cells overexpressing IRF1 and AML12 cells, the proportion of AML12 hepatocytes in late apoptosis was higher than that in the control group [(14.05±4.25) vs. (3.15±1.16), t=2.85, P=0.047], and the difference was statistically significant. In contrast, when the expression of IRF1 was knocked down, the proportion of apoptotic AML12 cells decreased [(9.26±3.04) vs. (13.36±4.64), t=2.15, P=0.098], but the difference was not statistically significant. Conclusion:The IRF1 protein can regulate the polarization of Kupffer cells into M1-type macrophages, promote the inflammatory injury of the liver tissue after ischemia-reperfusion, and increase the apoptosis of hepatocytes.

Objective To explore the effects of monocular deprivation(MD)on the expression of astrocytes in the superior colliculus,hippocampus,and visual cortex in mice during the critical period of visual development.Methods Eighteen C57BL/6J mice were randomly divided into the normal control group(CON group)and the MD group,with 9 mice in each group.Mice were bred under the 12 h/12 h dark/light conditions.Mice in the CON group received no treat-ment,while mice in the MD group underwent MD of the right eye on postnatal day 27,and the tissue was removed after 7 days.The mRNA and protein expression levels of glial fibrillary acidic protein(GFAP)in the superior colliculus,hippo-campus and visual cortex of mice in the two groups were detected using the real-time reverse transcription quantitative pol-ymerase chain reaction(RT-qPCR)and Western blot,respectively.The number of astrocytes labeled by GFAP and central nervous system specific protein β(S100β)in the superior colliculus,hippocampus and visual cortex of mice in the two groups was detected using the immunofluorescence staining.Results RT-qPCR and Western blot results showed that compared with the CON group,the mRNA and protein expression levels of GFAP in the superior colliculus,hippocampus(CA1,CA3 and dentate gyrus)and visual cortex of mice in the MD group decreased,and the differences were statistically significant(all P＜0.05).The immunofluorescence staining results showed that compared with the CON group,the number of GFAP and S1OOβ co-labeled astrocytes in the superior colliculus,hippocampus(CA1,CA3 and dentate gyrus)and visual cortex of mice in the MD group decreased,and the differences were statistically significant(all P＜0.05).Conclusion MD of mice during the critical period of visual development can result in a decrease in the number of astrocytes in the supe-rior colliculus,hippocampus and visual cortex.

Modulating Tankyrases (TNKS), interactions with USP25 to promote TNKS degradation, rather than inhibiting their enzymatic activities, is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway. Here, we identified UAT-B, a novel neoantimycin analog isolated from Streptomyces conglobatus, as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction (PPI) to overcome multi-drug resistance in colorectal cancer (CRC). The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels, triggering cell apoptosis through modulation of the Wnt/β-catenin pathway. Importantly, UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels, as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts, as well as APC^min/+ spontaneous CRC models. Collectively, these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment, and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.

Objective To reform the teaching content of acupuncture and moxibustion for undergraduate of Nursing College and to evaluate the effectiveness of the teaching reform,so to provide constructive information for optimizing the curriculum.Methods To revise the teaching content of Traditional Chinese Medicine(TCM)acu-puncture course based on the results of previous questionnaires,including reducing boring theoretical knowledge,increasing commonly used acupuncture techniques and clinical case analysis,organizing all undergraduates atten-ding the TCM teaching in the Nursing College of Peking Union Medical College to fill out the"Acupuncture Course Content Questionnaire"in 2023.The content covers four aspects:students'basic cognition of acupuncture and moxibustion before the course,students' learning feedback,students'learning effectiveness after the course and students' suggestions for further optimization of acupuncture and moxibustion course.Results Totally 149 students participated in the questionnaire survey and 73.15%of them were interested in the acupuncture course.There were 62.42%of the students thought that the most difficult part of the course was the"Acupuncture Points",and 71.81%of them complained the most constraining factor to the learning effectiveness was the diffi-culty of memorizing the meridians and acupoints.The most interesting part of the course was"Overview of Acu-puncture Treatment and Acupuncture Treatment of Common Diseases",which accounted for 44.3%of the students.Through the study,88.59%of the students were willing to recommend acupuncture treatment to pa-tients with indications.Students' suggestions for improvement of the acupuncture course was increase of classroom practice or learning by observation of operation videos.Conclusions The reformed acupuncture course highlights the practicability,improves the learning interest and subjective initiative,but the training of practice skill still needs to be strengthened.

In order to achieve large-scale production of HSV-IgM (HSV1, HSV2) human-mouse chimeric antibody in vitro, the gene sequence of the corresponding hybridoma cell was harvested by RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) technique to clone the chimeric antibody into eukaryotic expression vectors, and express the target proteins in CHO-S cells. At the same time, the screening process of stable cell lines was optimized, and the pressure conditions of pool construction stage and monoclonal screening stage were explored. Finally, the target protein was purified by protein L affinity purification method and the biological activity was detected. The recombinant IgM antibodies, HSV1 and HSV2, weighted at 899 kDa and 909 kDa respectively, were prepared. The optimal screening pressure was 20P200M (the first phase of pressure) and 50P1000M (the second phase of pressure). The final titer for the monoclonal expression of HSV1-IgM and HSV2-IgM was 1 620 mg/L and 623 mg/L, respectively. This study may facilitate the development of quality control products of HSV1 and HSV2 IgM series recombinant antibodies as well as efficient expression of IgM subtype antibodies in vitro.
Cricetinae ; Humans ; Animals ; Mice ; Immunoglobulin M/genetics* ; Antibodies, Viral ; CHO Cells ; Cricetulus ; Hybridomas ; Recombinant Fusion Proteins

Objective:To explore the feasibility and safety of robotic-assisted living donor left lateral segmentectomy (LDLLS) in a large pediatric liver transplant program.Methods:Retrospective analysis was performed for clinical data of 45 LDLLS donors and recipients from June 2021 to September 2022.Traditional open donor liver resection (n=30) and robotic-assisted segmentectomy (n=15) were performed.Two groups were compared with regards to operative duration, intraoperative hemorrhage, postoperative healing and postoperative complications.SPSS 21.0 was utilized for statistical analysis.Independent sample T, paired sample T, Wilcoxon rank sum and Chi-square tests were performed for examining the inter-group differences.Results:Operative duration of robot-assisted surgery group was substantially longer than that of traditional open surgery group ( P<0.001). Intraoperative blood loss was less in robot-assisted surgery group was less than that in traditional open surgery group[(106.0±39.8) vs.(251.0±144.8) ml, P=0.001]. Postoperative hospital stay of robot-assisted surgery group was shorter than that of traditional open surgery group[6.0(6.0, 6.0) vs.7.0(6.0, 9.0), P<0.05]. Two cases of postoperative biliary leakage were observed in donor of traditional open surgery group.Among 2 cases of abdominal infection, one was due to biliary leakage from liver section and secondary surgery was then performed.One case of incisional infection and another case of thrombosis occurred in donor of traditional open surgery group.In robot-assisted surgery group, only one donor had amylase elevation.In traditional open surgery group, there were one case of local thrombosis in middle hepatic vein and one case of bile duct stricture.No long-term complications occurred in robot-assisted surgery group during a follow-up period of over 6 months.Finally recipient data analysis indicated that no significant inter-group differences existed in operative duration, intraoperative blood loss, postoperative hospital stay or postoperative abdominal infection ( P=0.634, P=0.180, P=0.86 and P=0.153). Conclusions:Robotic-assisted LDLLS proves to be be a safe and reliable option for living donor segmentectomy.It is superior to conventional LDLLS in terms of shorter hospital stay, less intraoperative blood loss and fewer postoperative complications.