Objective:To predict the status of microvascular invasion (MVI) in intrahepatic cholangiocarcinoma (ICC) patients preoperatively based on the radiomics analysis of contrast-enhanced CT to provide imaging evidence for early identification of patients at high risk of recurrence.Methods:Clinical data of 40 ICC patients who underwent radical hepatectomy at Nanjing Drum Tower Hospital from January 2021 to May 2024 were retrospectively collected. Patients were divided into the MVI group ( n=8) and the non-MVI group ( n=32) according to the MVI status of the postoperative pathology report. Whether there were differences in each pathological index between the groups and the efficacy of radiomics analysis of contrast-enhanced CT for the preoperative prediction of MVI were analyzed. The regions of interest (ROI) were outlined on the arterial and venous phase images using the 3D Slicer software. Then, radiomics features were extracted from each ROI based on Python. Finally, the LASSO regression and glm function were used to screen radiomics features and establish a prediction model based on the R language. The established predictive model′s diagnostic efficacy, calibration, and net clinical benefit were evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA), respectively. Normally distributed measurement data were expressed as mean±standard deviation ( ± s) and compared using the t-test. Count data were expressed as frequency and compared using the chi-square test. Results:Patients in the MVI group had more poorly differentiated tumors and a significantly higher proportion of lymph node metastases ( P<0.05). The established radiomics prediction model included six features, 1 first-order statistical feature and 5 gray texture features. The area under the ROC curve was 0.87, the sensitivity was 75.0%, and the specificity was 90.6%. The calibration curve showed good agreement between the predicted MVI and actual MVI status, and the decision curve demonstrated that the model could provide a large net clinical benefit. Conclusion:Radiomics analysis of contrast-enhanced CT can identify the MVI status of ICC patients preoperatively and aid in clinical decision-making, providing vital evidence for individualized and precise treatment of ICC.

Objective:To investigate the effect and mechanism of the V-set and immunoglobulin domain-containing 4 ( VSIG4 ) gene mutation on the function of microglia in retinitis pigmentosa (RP). Methods:Localization of VSIG4 in the retina was detected by immunofluorescence.HMC3 cells (human microglial cells) were transfected with wild-type (Len-WT) VSIG4 gene, mutant type (Len-Mut) VSIG4 gene and empty vector virus (Len-Cont) and stimulated by the presence or absence of lipopolysaccharide (LPS), then divided into control group, LPS-Len-Mut group, LPS-Len-WT group, LPS-Len-Cont group, Len-Mut group, Len-WT group and Len-Cont group.The mRNA expression levels of the inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by real-time fluorescence quantitative PCR.Protein expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), glutathione peroxidase 4 (GPX4), nuclear transcription factor-κB (NF-κB) p65 subunit (P65), and phosphorylated P65 (PP65) were detected by Western blot.Cell phagocytic function was detected by phagocytosis assay.Cell migration ability was detected by cell scratch and transwell migration assay.LPS- stimulated HMC3 cells were co-cultured with 661W cells (mouse retinal photoreceptor cells), and the expression levels of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X (Bax) proteins of the cells were detected by Western blot.The number of apoptotic cells was determined by apoptosis assay. Results:VSIG4 was localized to microglia in mouse retina.The real-time fluorescence quantitative PCR results showed that compared with LPS-Len-WT group, the relative expression levels of IL-1β and TNF-α mRNA in HMC3 cells were significantly increased in LPS-Len-Mut group (both at P<0.05).The Western blot results showed that compared with LPS- Len-WT group, the protein expression levels of Nrf2, HO-1, and GPX4 in HMC3 cells were significantly reduced in LPS-Len-Mut group, and the PP65/P65 ratio was significantly increased (all at P<0.05).The phagocytic experiment results showed that the phagocytic rates of HMC3 cells in Len-Cont group, LPS-Len-Cont group, LPS-Len-WT group, and LPS-Len-Mut group were (35.67±3.22)%, (63.67±10.07)%, (84.00±3.46)%, and (64.67±2.31)%, respectively, showing a statistically significant difference ( F=59.06, P<0.001).Compared with LPS-Len-WT group, the phagocytic rate of HMC3 cells was significantly reduced in LPS-Len-Mut group ( P<0.05).The results of cell scratch and transwell migration assay showed that compared with LPS-Len-WT group, the migration rate of HMC3 cells at 24 and 48 hours and the number of invading cells per unit area at 24 hours were significantly reduced in LPS-Len-Mut group (all at P<0.05).Compared with LPS-Len-WT group, the expression ratio of Bax/Bcl-2 protein and the number of cell apoptosis were significantly increased in the LPS-Len-Mut group under the co-culture system (both at P<0.05). Conclusions:VSIG4 is localized to mouse retinal microglia.When the VSIG4 gene in RP mutates, HMC3 cells under LPS stimulation exhibit a series of changes, including activation of the NF-κB signaling pathway, decreased activation of the Nrf2/HO-1 signaling pathway, increased secretion of inflammatory cytokines, reduced phagocytic and migratory abilities, and increased cell apoptosis in co-culture systems.

Objective To establish a method for simultaneous determination of 14 components in Cinnamomi Cortex;To comprehensively evaluate its quality by multivariate statistical analysis technology.Methods HPLC method was used to establish the relative correction factors(RCF)between protocatechuic acid and 13 other components of coumalic acid,coumarin,cinnamyl alcohol,cinnamic acid,2-methoxycinnamic acid,cinnamaldehyde,o-methoxycinnamaldehyde,vanillic,syringic acid,4-hydroxybenzoic acid,procyanidin B2,epicatechin and procyanidin C1 to be measured,and the content of each component was calculated.The QAMS method was validated using external standard method(ESM),and the comprehensive quality of Cinnamomi Cortex was evaluated using PCA,OPLS-DA and EW-TOPSIS methods.Results The methodological verification results of ESM were in accordance with the requirements;the RCF established with protocatechuic acid as the reference had good durability under different experimental conditions,and the difference in content of each component between ESM and QAMS was not significant(P>0.05);the results of multivariate statistical analysis showed that accumulative contribution rate of former 2 principle components was 90.312%.Cinnamaldehyde,2-methoxycinnamic acid,procyanidin B2 and 4-hydroxybenzoic acid had a significant impact on the quality of Cinnamomi Cortex.The results of EW-TOPSIS showed that the Pi value of 16 batches of Cinnamomi Cortex ranged from 0.147 2 to 0.768 1.The quality of Cinnamomi Cortex from different producing areas varied greatly,and the overall quality of Cinnamomi Cortex from Guangxi Province was the best,followed by that from Guangdong Province and Yunnan Province,and the quality of Cinnamomi Cortex from Fujian Province was relatively poor.Conclusion The established QAMS multi-component quantitative method combined with multivariate statistical analysis technology can be used for comprehensive quality evaluation of Cinnamomi Cortex.

Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Humans ; Apolipoprotein E4/genetics* ; Cytosine ; Mutation ; Blastocyst ; Heterozygote ; Gene Editing ; CRISPR-Cas Systems

Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.

Objective:To investigate the risk factors for Gleason score upgrading after radical prostatectomy in clinical low-risk prostate cancer patients aged≥65 years.Methods:A total of 485 clinical low-risk prostate cancer patients aged≥65 years at five centers of the national multi-center PC-follow database from January 2015 to March 2019 were retrospectively analyzed.Data including age at diagnosis, prostate-specific antigen(PSA), MRI prostate imaging, puncture Gleason score, operation method, puncture method, positive incision margin and capsule penetration were collected.Differences in Gleason scores before and after operation were compared, and the risk factors for Gleason score upgrading after radical resection were evaluated by univariate and multivariate Logistic regression analysis.Results:Of 485 patients with a puncture Gleason score of 3+ 3=6, 261(53.8%)cases had postoperative pathological upgrading, in whom 228(87.4%)cases had Gleason score upgrading of 7, 22(8.4%)had Gleason score upgrading of 8, and 11(4.2%)had Gleason score upgrading of 9 or more.The rate of Gleason score upgrading was elevated with increased preoperative PSA levels, positive pelvic MRI, and higher positive rates of puncture biopsy.The incidences of postoperative capsule penetration(27.2% vs.12.5%, P<0.001)and positive incision margin(25.2% vs.17.4%, P=0.036)had statistically significant differences between the pathologically upgraded group and the pathologically non-upgraded group.Multivariate analysis showed that preoperative PSA level, percentage of positive puncture biopsies, biopsy Gleason score and pelvic MRI were independent predictors of prostate cancer. Conclusions:For clinical low-risk prostate cancer patients aged≥65 years with high risk factors for Gleason score upgrading, repeated biopsies should be carried out when necessary and the treatment plan should be adjusted accordingly.

Objective:To establish a nomogram model for predicting positive resection margins after prostate cancer surgery, and to perform the corresponding verification, in order to predict the risk of positive resection margins after surgery.Methods:A total of 2 215 prostate cancer patients from The First Affiliated Hospital of Naval Medical University, Hospital, Peking University First Hospital, Peking University Third Hospital, Peking University, and First Affiliated Hospital of Xi′an Jiaotong University were included in the PC-follow database from 2015 to 2018, and a simple random sampling method was used. They were divided into 1 770 patients in the modeling group and 445 patients in the verification group. In the modeling group, the age (<60 years, 60 to 70 years, >70 years), PSA (<4 ng/ml, 4-10 ng/ml, 11-20 ng/ml, >20 ng/ml), pelvic MRI (negative, suspicious, positive), clinical stage of the tumor (T 1-T 2, ≥T 3), percentage of positive needles (≤33%, 34%-66%, >66%), Gleason score of biopsy pathology (≤6 points, 7 points, ≥8 points). Univariate and multivariate logistic analysis were performed to screen meaningful indicators to construct a nomogram model. The model was used for validation in the validation group. Results:The results of multivariate analysis showed that preoperative PSA level ( OR=2.046, 95% CI 1.022 to 4.251, P=0.009), percentage of puncture positive needles ( OR=1.502, 95% CI 1.136 to 1.978, P=0.002), Gleason score of puncture pathology ( OR=1.568, 95% CI 1.063 to 2.313, P=0.028), pelvic MRI were correlated ( OR=1.525, 95% CI 1.160 to 2.005, P=0.033). Establish a nomogram model for independent predictors of positive margin of prostate cancer. The area under the receiver operating characteristic (ROC) curve of the validation group is 0.776. The area under the ROC curve of the preoperative PSA level, percentage of puncture positive needles, puncture pathology Gleason score, pelvic MRI, postoperative pathology Gleason score were 0.554, 0.615, 0.556, 0.522, and 0.560, respectively. The difference between the nomogram model and other indicators was statistically significant ( P<0.05). Conclusions:The constructed nomogram model has higher diagnostic value than the preoperative PSA level, percentage of puncture positive needles, Gleason score of puncturing pathology, pelvic MRI, and postoperative pathological Gleason score in predicting positive margin.

Objective To summarize the surgical skills of the pancreaticoduodenectomy via mesenteric approach for the treatment of duodenal gastrointestinal stromal tumors (GISTs).Methods The clinical data of 1 patient with huge duodenal GIST combined with gastrointestinal hemorrhage and liver metastasis who was admitted to the Second Affiliated Hospital of Harbin Medical University in June 2014 were retrospectively analyzed.The patient had continuous bleeding before the operation.The results of computed tomography (CT) showed that there was a solid tumor below the liver and in front of the right kidney,with the size of 12.2 cm × 8.1 cm,inferior vena cava was squeezed by the tumor,a solid tumor was detected in the right lobe of liver,and the colon was suspiciously invaded by the tumor.The patient received pancreaticoduodenectomy,right hemicolectomy and right lobectomy of liver metastases.The patient was followed up by out patient examination and telephone interview up to August 2014.Results The operation time,volume of blood loss and volume of red blood cell (RBC) transfusion were 420 minutes,800 mL and 2 U,respectively,with a full recovery of patients.The duodenal GIST with metastasis tumor and necrosis located at the colon,pancreatis,liver and renal hilum was confirmed by pathological diagnosis.The life quality of the patient was good by follow-up at postoperative month 3.Conclusion In the pancreaticoduodenectomy via mesenteric approach,surgeons perform vigorously the method of mesenteric approach at the first stage of operation,it could maximally reduce the risk of operation and the volume of blood loss.

Objective To evaluate the effects of sevoflurane on the development of posttraumatic stress disorder (PTSD) in rats and the role of hippocampal glycogen synthase kinase-3beta (GSK-3β).Methods Forty male Sprague-Dawley rats,weighing 250-300 g,aged 2 months,were randomly divided into 4 groups (n =10 each):control group (group C) ; PTSD group (group P) ; sevoflurane group (group S) ; lithium chloride (LiC1) + sevoflurane group (group LS).Establishment of PTSD model:the rats received 15 footshocks (1 mA,1 s shocks with a variable intershock interval of 240-480 s) in room A on the first day.On the second day,the rats received 1 footshock after an adaptation of 192 s in room B where its context was completely different from room A,and then they were removed from room B 32 s later.The rats were placed in room B and observed for 512 s on the third day.Group C did not receive any treatment on the first day in room A,while the other three groups received 15 footshocks in room A.In addition,0.8％ sevoflurane was administrated while training in group S,and group LS received intraperitoneal LiCl 100 mg/kg at 30 min before entering room A,and then the other treatments were similar to those previously described in group S.The ratio of freezing was recorded in room B on the second and third days.Four rats in each group were randomly sacrificed at 2 h after training in room A on the first day and their hippocampal tissues were taken to detect the expression of GSK-3β and phosphorylated GSK-3β (p-GSK-3β).Results Compared with group C,the ratio of freezing was significantly increased on the third day and the p-GSK-3β expression was up-regulated in P and LS groups,and no significant change was found in the parameters mentioned above in group S.Compared with group P,the ratio of freezing was significantly decreased on the third day and the p-GSK-3β expression was down-regulated in group S,and no significant change was found in the parameters mentioned above in group LS.Compared with group S,the ratio of freezing was significantly increased on the third day and the p-GSK-3β expression was up-regulated in group LS.There was no significant difference in the ratio of freezing on the second day and expression of p-GSK-3β between the four groups.Conclusion Sevoflurane can inhibit the development of PTSD in rats,and enhancement of hippocampal GSK-3β activity may be involved in the mechanism.