OBJECTIVE To evaluate the effectiveness and safety of warfarin anticoagulation therapy guided by gene test in patients undergoing left ventricular assist device （LVAD） implantation， and to analyze the influencing factors of warfarin anticoagulation efficacy. METHODS Patients who underwent LVAD implantation at the Heart and Vascular Center of Northern Jiangsu People’s Hospital from January 2023 to October 2024 and required warfarin anticoagulant therapy were selected as the study subjects. They were divided into genetic testing group （n＝51） and empirical treatment group （n＝17） based on whether they underwent CYP2C9 and VKORC1 gene test. The gene test group was given warfarin based on the predicted dose calculated by gene test， while the empirical treatment group was given warfarin by clinical doctors based on international normalized ratio （INR） experience， all patients were given warfarin once a day. Follow-up observation was conducted for 6 months to compare the effectiveness [time in therapeutic range（TTR）， the time required to reach INR for the first time， the incidence of embolic events， the incidence of INR＜1.5 events] and safety （the incidence of major and minor bleeding events，the incidence of INR＞3.5 events） of warfarin treatment between two groups of patients. According to whether the patient’s TTR was ≥60%， they were divided into TTR≥60% group （n＝20） and TTR＜60% group （n＝48）. Univariate and multivariate binary Logistic regression analysis were used to determine the factors affecting the anticoagulant effect of warfarin in patients. RESULTS The TTR of patients in the gene test group was significantly higher than that in the empirical treatment group （P＜0.05）. The incidence of INR＜1.5 events in the gene test group was significantly lower than in the empirical treatment group （P＜0.05）. The incidence of minor bleeding events and INR＞3.5 events in the gene test group were lower than in the empirical treatment group， but the difference was not statistically significant （P＞0.05）. The results of multivariate binary Logistic regression analysis showed that gene test was an independent protective factor for warfarin anticoagulant therapy [odds ratio （OR）＝10.842， 95% confidence interval （CI）： 1.211-27.037， P＝0.033]， and the combination of statins was an independent risk factor for warfarin anticoagulant therapy [OR＝0.196， 95%CI： 0.045-0.861， P＝0.031]. CONCLUSIONS Under the guidance of gene test， warfarin anticoagulation therapy for LVAD patients after implantation can improve TTR， shorten the anticoagulation target time， and has good safety； meanwhile， it should be noted that the combination of statins may enhance the anticoagulant effect of warfarin， thereby increasing the risk of bleeding in patients.

Objective:To comprehensively evaluate the performance of the iterative cone beam CT (iCBCT) imaging mode of Varian linear accelerators and to explore its specific advantages in clinical application.Methods:The kV cone beam CT (CBCT) imaging systems of Halcyon 2.0, Edge, and VitalBeam linear accelerators from Tianjin Medical University Cancer Institute & Hospital were selected, among which Halcyon 2.0 and Edge were equipped with the iCBCT imaging mode. The Penta-Guide phantom was used to evaluate the registration accuracy of iCBCT imaging modes. The accuracy of treatment couch position was measured by a ruler. The image quality of the iCBCT and conventional CBCT modes of various imaging devices were analyzed using the CatPhan604 phantom. The imaging beam-on time and reconstruction time were measured to assess image acquisition efficiency. The uniformity, spatial resolution, contrast, contrast-to-noise ratio (CNR), image acquisition time and reconstruction time between two imaging modes were statistically analyzed by t-test. Results:The maximum deviations of image registration measurement results of the iCBCT mode for Halcyon 2.0 and Edge accelerators compared to the standard values were 0.7 mm and 0.6 mm, respectively. The treatment couch position error of all devices was less than 1 mm. The iCBCT images under head scanning protocol primarily improved the uniformity and CNR. Compared to conventional CBCT images, Halcyon iCBCT increased the uniformity and CNR by 2.50% ( P<0.001) and 78.85% ( P<0.001), respectively, while Edge increased them by 2.18% ( P<0.001) and 86.42% ( P<0.001), both superior to VitalBeam CBCT images. Under pelvis scanning protocols, iCBCT images primarily improved the CNR compared to conventional CBCT images. Halcyon and Edge iCBCT increased the CNR by 113.57% ( P<0.001) and 133.87% ( P<0.001), respectively, both superior to VitalBeam CBCT images. In terms of image acquisition efficiency, the average reconstruction times for Halcyon and Edge iCBCT images increased by 7.28 s and 15.53 s, respectively, and the total image acquisition time of Halcyon accelerator was the shortest. Conclusions:While ensuring the registration accuracy, iCBCT imaging mode can significantly improve the CNR of images and improve the uniformity of images under head scanning protocol. The Halcyon imaging system can enhance image acquisition efficiency.

Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.

Objective:To summarize the clinical characteristics of patients with end-stage heart failure who receive heart transplant under extracorporeal membrane oxygenation (ECMO) support.Methods:The clinical data of 12 pediatric patients who received heart transplant with ECMO support in the Seventh Medical Center of Chinese People′s Liberation Army General Hospital and Guangdong Provincial People′s Hospital, from January 2019 to December 2023 was collected. The data included sex, age, weight, diagnosis, pre-ECMO lactate level, left ventricular ejection fraction (LVEF), vasoactive-inotropic score (VIS), and preoperative ECMO running time. Surgical data included cold ischemia time of the donor heart, cardiopulmonary bypass time, intraoperative use of immunosuppressant, postoperative use of ECMO, duration of postoperative ECMO, rate of successful weaning from ECMO, and survival discharge rate. The paired t-test was performed to compare cardiac function indices before and after left ventricular decompression. Results:The 12 patients ranged in age from 1.1 to 15.8 years, and weighted from 8 to 63 kg. Ten children were diagnosed with dilated cardiomyopathy, one with myocardial underdensification, and one with a novel heterozygous mutation of the SCN5A gene causing overlap syndrome complicated by fatal arrhythmia. Before ECMO, the lactate ranged from 0.6 to>15.0 mmol/L, the LVEF from 6.5% to 43%, and VIS from 3 to 108. Four patients underwent left ventricular decompression supported by preoperative ECMO, and their pulse pressure was significantly increased after decompression ((17.8±2.1) vs. (9.8±1.5) mmHg, 1 mmHg=0.133 kPa, t=11.31, P=0.001), while there was no apparent change in LVEF ((26.8±4.4)% vs. (24.9±4.9)%, t=1.75, P=0.178). A total of 7 children received a second run of ECMO after surgery and 3 of them successfully weaned off ECMO and survived to discharge. In the entire cohort, 10 were successfully weaned from ECMO and 8 survived to discharge. Conclusions:For children with end-stage heart failure supported by ECMO, left ventricular decompression can significantly improve pulse pressure. These patients will eventually require heart transplantation.

Objective To investigate the application effects of different myocardial protective solutions in total thoracoscopic minimally invasive aortic valve replacement surgery.Methods The clinical data of 72 patients with total thoracoscopic minimally invasive aortic valve replacement surgery in this hospital from May 2020 to January 2024 were analyzed retrospectively.The patients were divided into the St Thomas cardioplegia group(STH group,n=13),del Nido cardioplegia group(DN group,n=24),histidine tryptophan ketoglutar-ate solution group(HTK group,n=35)according to the different myocardial protective solutions.The levels of lactate(Lac)before and during surgery,the highest levels of myocardial creatine kinase isoenzyme(CK-MB),high-sensitivity troponin T(TnT)and creatinine(Cr)before operation,on the operative day and after surgery as well as the duration of extracorporeal circulation,aortic cross-clamping time,maximum flow rate,minimum bladder temperature,cardioplegia perfusion times,number of defibrillation after aortic de-clamping,postoperative ventilator assisted time,ICU stay duration and postoperative hospitalization duration were com-pared among the three groups.Results Except for 1 case of HTK was discharged automatically after surgery,the other 71 cases recovered and discharged according to the doctor's advice.There were no statistically signif-icant differences in the age,body weight,extracorporeal circulation time,aortic blocking time,maximum flow volume of extracorporeal circulation,minimum bladder temperature of extracorporeal circulation,Lac before extracorporeal circulation,highest Lac during extracorporeal circulation,assistant time of postoperative venti-lator,ICU stay duration,postoperative hospitalization duration,serum Cr before operation,Cr on operative day,preoperative TnT,postoperative TnT on operative day,postoperative highest TnT,preoperative CK-MB,postoperative CK-MB on operative day and postoperative highest CK-MB among the three groups(P＞0.05).There were statistically significant differences in the defibrillation ratio after aortic de-clamping and perfusion frequency of myocardial protective solution(P＜0.05).There was statistically significant difference in the perfusion frequency of myocardial protective solution in pairwise comparison among groups(P＜0.05),and the defibrillation ratio after aortic de-clamping had statistical difference between the DN group and HTK group(P＜0.05).Conclusion DN,STH and HTK all have good myocardial protective effect in total thoraco-scopic minimally invasive aortic valve surgery.HTK has the advantages of less perfusion times and decreasing the operative procedures compared with DN and STH;DN has the advantage of lower use for electrical defib-rillation correcting arrhythmias after aortic opening over HTK.

Objective:To compare the efficacy of 3D-printed guide plate assisted versus freehand placement of cannulated screws for the treatment of Sanders type II and III calcaneal fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 29 patients with Sanders type II and III calcaneal fractures admitted to Chonggang General Hospital from June 2020 to October 2022. Among them, there were 18 males and 11 females, with an age range of 22-69 years [(40.1±11.5)years]. Nineteen patients were treated with individualized 3D-printed guide plate assisted placement of cannulated screws (3D-printed group) and 10 were treated with freehand placement of cannulated screws (freehand group). The surgical time, fluoroscopy times, postoperative 6-month calcaneal morphology (length, width, height, B?hler angle and Gissane angle), and American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Maryland functional score assessed at 3, 6 months after operation and at the final follow-up were compared between the two groups. The incision healing and complications were observed.Results:The patients were followed up for 6-24 months [(11.3±2.5)months]. The surgical time and fluoroscopy times in the 3D-printed group were (53.4±9.1)minutes and (7.3±1.1)times, respectively, which were shorter than (90.2±16.0)minutes and (16.0±3.2)times in the freehand group (all P<0.01). At 6 months after operation, there was no significant difference in calcaneal length between the two groups ( P>0.05); the calcaneal width, height, B?hler angle and Gissane angle in the 3D-printed group [(34.0±1.8)mm, (47.2±1.6)mm, (27.8±1.0)°, (129.2±2.8)°] were superior than those in the freehand group [(37.5±2.0)mm, (43.0±2.7)mm, (25.8±1.5)°, (125.9±2.5)°] (all P<0.01). At 3, 6 months after operation and at the final follow-up, the values of AOFAS ankle-hindfoot score in the 3D-printed group [(72.2±2.3)points, (79.7±2.3)points, (86.5±4.4)points] were higher than those in the freehand group [(64.2±6.9)points, (73.4±4.2)points, (81.8±3.1)points] (all P<0.05); the values of Maryland score in the 3D-printed group [(71.4±7.7)points, (84.7±2.6)points, (91.5±2.5)points] were higher than those in the freehand group [(65.2±5.6)points, (79.1±3.8)points, (87.1±2.9)points] (all P<0.05). All surgical incisions were healed in stage I. In the 3D-printed group, there were no complications regarding infection, iatrogenic vascular or nerve injury, or fixation failure after surgery. In the freehand group, one patient with lateral sural cutaneous nerve injury was resolved spontaneously without specific treatment. Conclusion:Compared with freehand placement of cannulated screws, 3D-printed guide plate assisted placement of percutaneous placement has the advantages of shorter surgical time, fewer fluoroscopy times, lower reduction loss, better ankle joint function recovery, and less complications in treating Sanders type II and III calcaneal fractures.

Objective:A high-throughput assay for the detection of five common clinical Candidaemia pathogens was established by combining polymerase chain reaction (PCR) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).Method:Establishment of methodology. We selected Candida albicans, Candida parapsilosis, Candida glabrata, Candida krusei, Candida tropicalis to be the target pathogens and the internal transcribed spacer (ITS) region as the target gene. Specific single base extension primers were designed to perform single base extension reaction in the same reaction system. MALDI-TOF MS was used to detect the characteristic peaks of each target pathogen. The sensitivity and specificity of the detection system were verified by using spiked blood samples. Totally 108 blood samples from proven or suspected candidaemia patients were collected from October 2021 to September 2022 in a hospital in Beijing. The results of nucleic acid mass spectrometry were compared with those of clinical blood culture. Results:The established nucleic acid mass spectrometry detection system can simultaneously detect five common clinical Candida species. Each strain can produce specific product peaks and there is no mutual interference between the strains. The detection limit of Candida albicans was 100 CFU/ml. The detection limit of Candida parapsilosis, Candida glabrata, Candida krusei and Candida tropicalis was 10 CFU/ml. For the 108 blood samples, the sensitivity, specificity, positive predictive value and negative predictive value of nucleic acid mass spectrometry were 94.74% (36/38), 97.14% (68/70), 92.31% (36/39) and 98.55% (68/69), respectively. The McNemar χ 2 test showed no significant difference between the two methods ( P>0.05), and the Kappa consistency test showed good consistency between the two methods ( Kappa=0.9, P<0.05). Conclusion:A nucleic acid mass spectrometry detection system suitable for clinical candida detection was successfully constructed, and the method validation results were consistent with the clinical blood culture.

Objective:To investigate the clinical efficacy of proximal gastrectomy and total gastrectomy in the treatment of Siewert type Ⅱ and Ⅲ adenocarcinoma of esophagogastric junction (AEG).Methods:The retrospective cohort study was conducted. The clinicopathological data of 170 patients with Siewert type Ⅱ and Ⅲ AEG who were admitted to Guangdong Provincial People′s Hospital from January 2010 to December 2018 were collected. There were 125 males and 45 females, aged from 30 to 85 years, with a median age of 64 years. Of the 170 patients, 82 cases undergoing proximal gastrectomy were allocated into the proximal gastrectomy group and 88 cases undergoing total gastrectomy were allocated into the total gastrectomy group. Observation indica-tors: (1) surgical and postoperative situations; (2) follow-up and survival; (3) analysis of prognostic factors. Follow-up was conducted using telephone interview and outpatient examination to detect survival of patients up to December 2021. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measure-ment data with skewed distribution were represented as M( Q1, Q3) or M(range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Comparison of ordinal data was analyzed using the rank sum test. Kaplan-Meier method was used to draw survival curves, and Log-Rank test was used for survival analysis. COX proportional hazard model was used for univariate and multivariate analyses. Variables with P<0.1 in univariate analysis were included for multivariate analysis. Results:(1) Surgical and postoperative situations. Cases with surgical approach as transthoracic or thoraco-abdominal approach, transabdominal approach, the operation time, cases with volume of intra-operative blood loss ≤100 mL or >100 mL, cases with length of proximal margin ≤1.5 cm or >1.5 cm, cases with radical surgery outcome as R 0, R 1, R 2, the number of lymph nodes harvest, cases with anastomotic leakage, cases with anastomotic stricture, cases with incision infection, cases with pleural infection or effusion, cases with abdominal infection or ascites were 61, 21, (211±18)minutes, 46, 36, 44, 38, 73, 6, 3, 15(9,22), 5, 2, 2, 4, 2 in the proximal gastrectomy group, respec-tively. The above indicators were 12, 76, (263±15)minutes, 27, 61, 45, 43, 82, 4, 2, 23(18,32), 4, 1, 3, 1, 4 in the total gastrectomy group, respectively. There were significant differences in the surgical approach, operation time, volume of intraoperative blood loss and the number of lymph nodes harvest between the two groups ( χ2=63.94, t=-25.50, χ2=11.19, Z=-5.62, P<0.05). There was no significant difference in the length of proximal margin or radical surgery outcome between the two groups ( χ2=0.11, Z=-0.95, P>0.05) and there was no significant difference in the anastomotic leakage, anastomotic stricture, incision infection, pleural infection or effusion, abdominal infection or ascites between the two groups ( P>0.05). (2) Follow-up and survival. All the 170 patients were followed up for 89(64,106)months. Of the 170 patients, the 5-year overall survival rates were 43.8% and 35.5% of the Siewert type Ⅱ and Ⅲ AEG patients, respectively, showing no significant difference between them ( χ2=0.87, P>0.05). Of the patients with Siewert type Ⅱ AEG, the 5-year overall survival rates were 41.7% and 54.3% in the patients with proximal gastrectomy and the total gastrectomy, respectively, showing no significant difference between them ( χ2=1.05, P>0.05). Of the patients with Siewert type Ⅲ AEG, the 5-year overall survival rates were 31.3% and 37.5% in the patients with proximal gastrectomy and the total gastrectomy, respectively, showing no significant difference between them ( χ2=0.33, P>0.05). The 5-year overall survival rates were 39.0% and 44.2% in the proximal gastrectomy group and the total gastrectomy group, respectively, showing no significant difference between the two groups ( χ2=0.63, P>0.05). Of the patients in TNM stage Ⅰ, stage Ⅱ, stage Ⅲ, the 5-year overall survival rates were 65.3%, 36.3%, 27.1% in the proximal gastrectomy group, versus 83.3%, 48.0%, 39.7% in the total gastrectomy group, showing no signifi-cant difference between the two groups ( χ2=0.02, 1.50, 1.21, P>0.05). (3) Analysis of prognostic factors. Results of univariate analysis showed that pathological N staging, degree of tumor differen-tiation and radical surgery outcome were related factors influencing prognosis of AEG patients ( hazard ratio=1.71, 1.70, 2.85, 95% confidence interval as 1.16-2.60, 1.15-2.50, 1.58-5.14, P<0.05). Results of multivariate analysis showed that pathological N staging and radical surgery outcome were independent factors influencing prognosis of AEG patients ( hazard ratio=1.55, 2.18, 95% confidence interval as 1.05-2.31, 1.18-4.02, P<0.05). Conclusions:There is no significant difference in the prognosis of Siewert type Ⅱ and Ⅲ AEG patients undergoing proximal gastrectomy or total gastrectomy. Proximal gastrectomy can be used for the treatment of advanced Siewert type Ⅱ and Ⅲ AEG.

Objective:By using balloon occlusive hepatic angiography in cirrhotic portal hypertension to evaluate contrast doses on the detection rate of intrahepatic venous-lateral branch shunt (HVVC), and the effect on hepatic venous pressure gradient (HVPG) and portal vein pressure gradient (PPG).Methods:From Jan 2018 to Jun 2021, 131 patients received transjugular intrahepatic portosystemic shunt (TIPS) at Beijing Shijitan Hospital.Results:A positive correlation between PVP and weged hepatic venous pressure (WHVP) ( r=0.241, P=0.001) was found when only by right hepatic vein approach. Ten ml of iodine contrast medium when compared to 5ml doses found more cases of intrahepatic venous-venous lateral branch shunt. The mean PPG of patients with HVVC was significantly higher than the mean of HVPG( P<0.05).The right hepatic vein was the only reliable vein by which WHVP was measured. Conclusions:Right hepatic vein manometry,adequate ballon occlusion and using 10ml of iodine contrast help get reliable WHVP and found HVVC; HVVC can affect the consistency of HVPG and PPG.

SAM pointed domain containing E26 transformation-specific transcription factor (SPDEF) plays dual roles in the initiation and development of human malignancies. However, the biological role of SPDEF in head and neck squamous cell carcinoma (HNSCC) remains unclear. In this study, the expression level of SPDEF and its correlation with the clinical parameters of patients with HNSCC were determined using TCGA-HNSC, GSE65858, and our own clinical cohorts. CCK8, colony formation, cell cycle analysis, and a xenograft tumor growth model were used to determine the molecular functions of SPDEF in HNSCC. ChIP-qPCR, dual luciferase reporter assay, and rescue experiments were conducted to explore the potential molecular mechanism of SPDEF in HNSCC. Compared with normal epithelial tissues, SPDEF was significantly downregulated in HNSCC tissues. Patients with HNSCC with low SPDEF mRNA levels exhibited poor clinical outcomes. Restoring SPDEF inhibited HNSCC cell viability and colony formation and induced G0/G1 cell cycle arrest, while silencing SPDEF promoted cell proliferation in vitro. The xenograft tumor growth model showed that tumors with SPDEF overexpression had slower growth rates, smaller volumes, and lower weights. SPDEF could directly bind to the promoter region of NR4A1 and promoted its transcription, inducing the suppression of AKT, MAPK, and NF-κB signaling pathways. Moreover, silencing NR4A1 blocked the suppressive effect of SPDEF in HNSCC cells. Here, we demonstrate that SPDEF acts as a tumor suppressor by transcriptionally activating NR4A1 in HNSCC. Our findings provide novel insights into the molecular mechanism of SPDEF in tumorigenesis and a novel potential therapeutic target for HNSCC.
Carcinogenesis ; Cell Proliferation ; Head and Neck Neoplasms ; Humans ; Nuclear Receptor Subfamily 4, Group A, Member 1 ; Proto-Oncogene Proteins c-ets ; Squamous Cell Carcinoma of Head and Neck ; Transcription Factors