Objective To analyze the clinical efficacy of valve surgeries for infective endocarditis and the affecting factors, and compare the early- and long-term postoperative outcomes of different surgery approaches. Methods The patients with infective endocarditis who underwent valve replacement/valvuloplasty in our hospital from 2010 to 2022 were retrospectively collected. The clinical data of the patients were analyzed. Results A total of 343 patients were enrolled, including 197 patients with mechanical valve replacement, 62 patients with bioprosthetic valve replacement, and 84 patients with valvuloplasty. There were 238 males and 105 females with an average age of (44.2±14.8) years. Single-valve endocarditis was present in 200 (58.3%) patients, and multivalve involvement was present in 143 (41.7%) patients. Sixty (17.5%) patients had suffered thrombosis before surgery, including cerebral embolisms in 32 patients. The mean follow-up time was (60.6±43.8) months. Early mortality within one month after the surgery occurred in 17 (5.0%) patients, while later mortality occurred in 19 (5.5%) patients. Eight (2.3%) patients underwent postoperative dialysis, 13 (3.8%) patients suffered postoperative stroke, 6 patients underwent reoperation, and 3 patients suffered recurrence of infective endocarditis. Smoking (P=0.002), preoperative embolisms (P=0.001), duration of surgery (P=0.001), and postoperative dialysis (P=0.001) were risk factors for early mortality, and left ventricular ejection fraction ≥60% (P=0.022) was protective factor for early mortality. New York Heart Association classification Ⅲ-Ⅳ (P=0.010) and ≥3 valve procedures (P=0.028) were risk factors for late mortality. The rate of composite endpoint events was significantly lower in the valvuloplasty group than that in the valve replacement group. Conclusion For patients with infective endocarditis, smoking and preoperative embolisms are associated with high postoperative mortality, multiple-valve surgery is associated with a poorer prognosis, and valvuloplasty has advantages over valve replacement and should be attempted in the surgical management of patients with infective endocarditis.

Objective:To prepare IL-12 modified with polyethylene glycol derivatives and determine its modification sites,and to evaluate it in vitro in terms of stability,hemogram recovery,anti-tumor effects and other aspects.Methods:IL-12 was modified with polyethylene glycol propionaldehyde and maleimide imide derivatives,and modified sites of two different derivatives were deter-mined by ultra performance liquid chromatography(UPLC).Recombinant human IL-12 and PEG-IL-12 were used to stimulate NK92 cells,activity and cytotoxicity of IFN-γ were evaluated by kits.CD34+was stimulated to evaluate its blood picture recovery potential.NK cell killing was stimulated to evaluate its effectiveness in improving immunity and anti-tumor effects.Results:Compared with recombinant human IL-12,PEG-IL-12 had higher stability,blood picture recovery potential and anti-tumor effects.Conclusion:PEG-IL-12 can effectively overcome many drawbacks of recombinant human IL-12,greatly improving possibility of its widespread applica-tion in clinical trials.

Objective:To compare and discuss the mid-to-long-term outcomes of mitral valve repair (MVP) versus biological mitral valve replacement (bMVR) in patients aged 60 years and above with rheumatic mitral valve disease.Methods:This is a retrospective cohort study. A total of 765 patients aged 60 years and older, diagnosed with rheumatic mitral valve disease and who underwent MVP or bMVR at Beijing Anzhen Hospital from January 2010 to January 2023, were retrospectively included. Among them, 186 were male and 579 were female, with an age of (66.1±4.5) years (range: 60 to 82 years). Patients were divided into two groups based on the surgical method: the mitral valve repair group (MVP group, n=256) and the bioprosthetic mitral valve replacement group (bMVR group, n=509). A 1∶1 propensity score matching was performed using a caliper value of 0.2 based on preoperative data. Paired sample t-tests, χ2 tests, or Fisher′s exact tests were used for intergroup comparisons. Kaplan-Meier method was employed to plot survival curves and valve-related reoperation rate curves for both groups before and after matching, and Log-rank tests were used to compare the mid-to long-term survival rates and valve-related reoperation rates between the two groups. Results:A total of 765 patients who completed follow-up were ultimately included, with a follow-up period ( M(IQR)) of 5.1(5.0) years (range: 1.0 to 12.9 years). After matching, each group consisted of 256 patients. The incidence of early postoperative atrial fibrillation (39.1% vs. 49.2%, χ2=4.95, P=0.026) and early mortality rates (2.0% vs. 6.2%, χ2=4.97, P=0.026) were lower in the MVP group. Unadjusted Kaplan-Meier analysis showed significantly higher 5-year and 10-year survival rates for the MVP group (92.54% vs. 83.02%, 86.22% vs. 70.19%, Log-rank: P=0.001). After adjustment with propensity scores, the Kaplan-Meier analysis still indicated higher 5-year and 10-year survival rates in the MVP group compared to the bMVR group (92.54% vs. 85.89%, 86.22% vs. 74.83%, Log-rank: P=0.024). There were no significant differences in the rates of valve-related reoperation between the two groups before and after matching (5-year and 10-year reoperation rates pre-matching: 1.75% vs. 0.57%, 5.39% vs. 7.54%, Log-rank: P=0.207; post-matching: 1.75% vs. 0, 5.39% vs. 9.27%, Log-rank: P=0.157). Conclusion:For patients aged 60 years and above with rheumatic mitral valve disease, mitral valve repair offers better mid-to-long-term survival compared to biological valve replacement.

Objective:To compare and discuss the mid-to-long-term outcomes of mitral valve repair (MVP) versus biological mitral valve replacement (bMVR) in patients aged 60 years and above with rheumatic mitral valve disease.Methods:This is a retrospective cohort study. A total of 765 patients aged 60 years and older, diagnosed with rheumatic mitral valve disease and who underwent MVP or bMVR at Beijing Anzhen Hospital from January 2010 to January 2023, were retrospectively included. Among them, 186 were male and 579 were female, with an age of (66.1±4.5) years (range: 60 to 82 years). Patients were divided into two groups based on the surgical method: the mitral valve repair group (MVP group, n=256) and the bioprosthetic mitral valve replacement group (bMVR group, n=509). A 1∶1 propensity score matching was performed using a caliper value of 0.2 based on preoperative data. Paired sample t-tests, χ2 tests, or Fisher′s exact tests were used for intergroup comparisons. Kaplan-Meier method was employed to plot survival curves and valve-related reoperation rate curves for both groups before and after matching, and Log-rank tests were used to compare the mid-to long-term survival rates and valve-related reoperation rates between the two groups. Results:A total of 765 patients who completed follow-up were ultimately included, with a follow-up period ( M(IQR)) of 5.1(5.0) years (range: 1.0 to 12.9 years). After matching, each group consisted of 256 patients. The incidence of early postoperative atrial fibrillation (39.1% vs. 49.2%, χ2=4.95, P=0.026) and early mortality rates (2.0% vs. 6.2%, χ2=4.97, P=0.026) were lower in the MVP group. Unadjusted Kaplan-Meier analysis showed significantly higher 5-year and 10-year survival rates for the MVP group (92.54% vs. 83.02%, 86.22% vs. 70.19%, Log-rank: P=0.001). After adjustment with propensity scores, the Kaplan-Meier analysis still indicated higher 5-year and 10-year survival rates in the MVP group compared to the bMVR group (92.54% vs. 85.89%, 86.22% vs. 74.83%, Log-rank: P=0.024). There were no significant differences in the rates of valve-related reoperation between the two groups before and after matching (5-year and 10-year reoperation rates pre-matching: 1.75% vs. 0.57%, 5.39% vs. 7.54%, Log-rank: P=0.207; post-matching: 1.75% vs. 0, 5.39% vs. 9.27%, Log-rank: P=0.157). Conclusion:For patients aged 60 years and above with rheumatic mitral valve disease, mitral valve repair offers better mid-to-long-term survival compared to biological valve replacement.

During the gene editing process mediated by CRISPR/Cas9, precise genome editing and gene knock-in can be achieved by the homologous recombination of double-stranded DNA (dsDNA) donor template. However, the low-efficiency of homologous recombination in eukaryotic cells hampers the development and application of this gene editing strategy. Here, we developed a novel CRISPR/Cas9-hLacI donor adapting system (DAS) to enhance the dsDNA-templated gene editing, taking the advantage of the specific binding of the LacI repressor protein and the LacO operator sequence derived for the Escherichia coli lactose operon. The codon-humanized LacI gene was fused as an adaptor to the Streptococcus pyogenes Cas9 (SpCas9) and Staphylococcus lugdunensis Cas9 (SlugCas9-HF) genes, and the LacO operator sequence was used as the aptamer and linked to the dsDNA donor template by PCR. The Cas9 nuclease activity after the fusion and the homology-directed repair (HDR) efficiency of the LacO-linked dsDNA template were firstly examined using surrogate reporter assays with the corresponding reporter vectors. The CRISPR/Cas9-hLacI DASs mediated genome precise editing were further checked, and we achieved a high efficiency up to 30.5% of precise editing at the VEGFA locus in HEK293T cells by using the CRISPR/SlugCas9-hLacI DAS. In summary, we developed a novel CRISPR/Cas9-hLacI DAS for dsDNA-templated gene editing, which enriches the CRISPR/Cas9-derived gene editing techniques and provides a novel tool for animal molecular design breeding researches.
Humans ; Animals ; Gene Editing ; CRISPR-Cas Systems/genetics* ; HEK293 Cells ; Homologous Recombination ; DNA

OBJECTIVE To investigate the effects of the compo nents of traditional Chinese medicine （TCM）（total ginsenoside，total ketone ester of Ginkgo biloba ，total glucoside of Crocus sativus ）for invigorating qi and activating blood circulation on cognitive function of mice with chronic cerebral ischemia （CCI）and its related mechanisms . METHODS CCI model was established by stimulating bilateral common carotid arteries of C 57BL/6J mice with temperature controlled current . Model mice were randomly divided into model group ，aspirin group （positive control 10 mg/kg）and TCM group （the components of TCM for invigorating qi and activating blood circulation 33 mg/kg），sham operation group was set up additionally ，with 16 mice in each group. Except that the mice in model group and sham operation group were given water intragastrically ，the mice in the other groups were given corresponding medicine liquid intragastrically，once a day ，for 8 weeks. The latency of step down and the number of step -down errors of mice in each group were detected ；the ultrastructure of neurovascular units（NVU）in cerebral cortex and hippocampus was observed ；the protein and mRNA expressions of vascular endothelial growth factor（VEGF）and α7 nicotinic acetylcholine receptors （α7nAChRs） in cerebral tissue were detected ；the contents of VEGF，angiopoietin 1 （Ang1） and basic fibroblast growth factor（bFGF）were detected . RESULTS Compared with model group ，the latency of step -down was significantly prolonged （P＜ 0.05），and the number of step -down errors was significantly reduced （P＜0.05）in TCM group . The ultrastructural abnormalities of NVU in cerebral cortex and hippocampus were significantly improved ，and the protein and mRNA expressions of VEGF and α7 nAChRs and the contents of VEGF ，Ang1 and bFGF in cerebral tissue were significantly increased （P＜0.05）. CONCLUSIONS The components of TCM for invigorating qi and activating blood circulation can improve CCI -induced cognitive dysfunction in mice，the mechanism of which may be associated with protecting NVU ，recovering ultrastructural abnormalities of injured NVU ， and regulating the expression of proteins closely related to NVU and memory function .

OBJECTIVE: To investigate the effect of miR-186 inhibition on the expression of hypoxia-inducible factor-1α (HIF-α) and mitochondrial function in hypoxic vascular endothelial cells. METHODS: Human umbilical vein endothelial cells (HUVECs) cultured in routine or hypoxic conditions for 6 h were examined for the expression of miR-186. A miR-186 inhibitor was transfected in the HUVECs, and the cells were subsequently cultured in hypoxic condition for 6 h to observe the changes in the mitochondrial structure under an electron microscope. The changes in the mRNA and protein expressions of HIF-1α in response to miR-186 interference were tested using real-time fluorescent quantitative PCR and Western blotting. RESULTS: The expression of miR-18 was mildly increased in HUVECs after hypoxic exposure for 6 h (=0.0188). Interference of miR-186 expression obviously promoted the mRNA and protein expressions of HIF-1α in HUVECs. In hypoxic conditions, miR-186 interference significantly reduced mitochondrial damage in HUVECs as observed under electron microscope (=0.0297). CONCLUSIONS Inhibition of miR-186 protects vascular endothelial cells against hypoxic injuries by promoting HIF-α expression to lessen mitochondrial damage, suggesting the possibility of targeted miR-186 interference for treatment of hemorrhagic shock.
Cell Hypoxia ; Human Umbilical Vein Endothelial Cells ; Humans ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit ; MicroRNAs ; Mitochondria ; Umbilical Veins

Objective    To compare long-term outcomes following mitral valvuloplasty (MVP) and mitral valve replacement (MVR) for native valve endocarditis (NVE). Methods    Between November 1993 and August 2016, consecutive 101 patients with NVE underwent mitral surgery in our department, MVP for 52 patients and MVR for 49 patients. There were 69 males and 32 females at age of 38.1±14.9 years. The mean follow-up was 99.4±75.8 months. Results    There was no statistical difference in cardiopulmonary bypass time, aortic cross-clamp time, in-hospital mortality, duration of mechanical ventilation, ICU stay or hospital stay after surgery between the two groups. Survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 97.6%, 97.6%, 97.6% for MVP, and 93.5%, 84.3%, 84.3%, 66.2% for MVR with a statistical difference between the two groups (P=0.018). There was no stroke in the patients with MVP during follow-up periods. However, stroke-free survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 93.9%, 89.4%, 70.2% for MVR patients with a statistical difference between the two groups (P=0.023). There was no statistical difference in recurrence of infection, perivalvular leakage and reoperation between the two groups. Composite endpoint-free survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 97.6%, 92.9%, 92.9% for MVP, and 91.3%, 79.6%, 75.8%, 51.0% for MVR with a statistical difference (P=0.006). Conclusion    MVP is associated with better outcomes than MVR in the patients  with NVE; generalizing MVP technique in the patients with NVE is needed.

OBJECTIVETo investigated the characteristics and regularity of lymph node metastasis around gastric cancer in order to provide reference for standardized and optimal surgical treatment.

METHODSA retrospective case series study was carried out on 1456 patients with gastric cancer undergoing surgical treatment at the Affiliated Tumor Hospital of Tianjin Medical University from March 2003 to August 2011. The number of harvested lymph node and metastasis status of various lymph node station were determined by routine pathological examination of specimens, including resected gastric tissue and dissected lymph node tissue, according to the 13th version of the Japanese Gastric Cancer Treatment Guidelines. Tumor T, N and M staging was performed to evaluate lymphatic metastasis status of different locations of gastric cancer according to the TNM staging criteria of the 8th edition of the American Joint Committee on Cancer (AJCC). The influence of gender, age, tumor diameter, Borrmann type, T staging and M staging, tumor differentiation degree, invasion of vessels, lymphatic vessels and nerves, radical surgical degree and other clinical factors on lymph node metastasis was analyzed.

RESULTSA total of 1062 cases(72.9%) had lymph node metastasis in the 1456 patients with gastric cancer. A total of 9766 lymph nodes were positive for metastasis. Lymph node metastasis occurred in 11 of 44 (25.0%) patients with early gastric cancer and in 1051 of 1412 (74.4%) patients with advanced gastric cancer. The largest number of lymph node metastases was found in No.3 station [653 cases (44.8%)], followed by No.6 [437 cases(30.0%)], No.7 [345 cases (23.7%)], No.1 [304 cases (20.9%)], No.4sb [290 cases (19.9%)]. No.14v lymph node metastasis was observed in 23 cases, of whom No.6 (16 cases, 69.6%), No.8a (15 cases, 65.2%) and No.3 (12 cases, 52.2%) developed simultaneous metastasis. As for different locations of gastric cancer, stations with more lymph node metastasis in 309 patients with proximal gastric cancer were No.3 (133 cases, 43.0%), No.1 (96 cases, 31.1%), No.2 (90 cases, 29.1%) and No.7 (89 cases, 28.8%); in 144 patients with middle gastric cancer were No.3 (68 cases, 47.2%), No.6 (50 cases, 34.7%), No.7 (40 cases, 27.8%) and No.4sb (38 cases, 26.4%); in 700 patients with distal gastric cancer were No.3(287 cases, 41.0%), No.6 (265 cases, 37.8%), No.4sb (138 cases, 19.7%) and No.8a (138 cases, 19.7%); in 303 cases with diffuse-type gastric cancer were No.3 (165 cases, 54.4%), No.6 (100 cases, 33.0%), No.7 (88 cases, 29.0%), No.1 (84 cases, 27.7%) and No.4sb (72 cases, 23.8%). The incidence of lymph node skip metastasis was 7.2% (105/1456) in whole group. Positive lymph node metastasis was associated with tumor size (RR=2.016, 95%CI: 1.550-2.621, P=0.000), tumor differentiation(RR=1.631, 95%CI:1.405-1.894, P=0.000), tumor T staging (RR=1.886, 95%CI: 1.629-2.184, P=0.000), tumor M staging (RR=3.671, 95%CI:1.265-10.660, P=0.017) and radical surgery(RR=3.819, 95%CI: 2.023-7.207, P=0.000).

CONCLUSIONSThe main direction of peripheral lymph node drainage in gastric cancer is lesser curvature, and then the left gastric artery, the common hepatic artery and the peripheral lymph nodes of the celiac axis, and finally the peripheral lymph nodes of the abdominal aorta. Therefore the No.6 station lymph node adjacent to the perigastric area, the No.7, No.8 and No.9 lymph nodes should be the focus of the radical surgical dissection of gastric cancer. Tumor size, differentiation degree, invasion depth and distant metastasis have significant association with lymph node metastasis. For patients with adverse factors, radical surgery is necessary to ensure efficacy.