Donors with a serum sodium concentration of >155 mmol/L are extended criteria donors for liver transplantation (LT). Elevated serum sodium of donors leads to an increased incidence of hepatic dysfunction in the early postoperative period of LT; however, the exact mechanism has not been reported. We constructed a Lewis rat model of 70% hepatic parenchymal area subjected to ischemia-reperfusion (I/R) with hypernatremia and a BRL-3A cell model of hypoxia-reoxygenation (H/R) with high-sodium (HS) culture medium precondition. To determine the degree of injury, biochemical analysis, histological analysis, and oxidative stress and apoptosis detection were performed. We applied specific inhibitors of the epithelial sodium channel (ENaC) and Na⁺/Ca²⁺ exchanger (NCX) in vivo and in vitro to verify their roles in injury. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) levels and the area of hepatic necrosis were significantly elevated in the HS+I/R group. Increased reactive oxygen species (ROS) production, myeloperoxidase (MPO)‍-positive cells, and aggravated cellular apoptosis were detected in the HS+I/R group. The HS+H/R group of BRL-3A cells showed significantly increased cellular apoptosis and ROS production compared to the H/R group. The application of amiloride (Amil), a specific inhibitor of ENaC, reduced ischemia-reperfusion injury (IRI) aggravated by HS both in vivo and in vitro, as evidenced by decreased serum transaminases, inflammatory cytokines, apoptosis, and oxidative stress. SN-6, a specific inhibitor of NCX, had a similar effect to Amil. In summary, hypernatremia aggravates hepatic IRI, which can be attenuated by pharmacological inhibition of ENaC or NCX.
Animals ; Reperfusion Injury/drug therapy* ; Hypernatremia/complications* ; Rats ; Liver/metabolism* ; Rats, Inbred Lew ; Male ; Apoptosis ; Sodium-Calcium Exchanger/antagonists & inhibitors* ; Reactive Oxygen Species/metabolism* ; Oxidative Stress ; Epithelial Sodium Channel Blockers/pharmacology* ; Epithelial Sodium Channels ; Cell Line ; Liver Transplantation

Objective To generate a dense granule protein 3 (GRA3) gene-deficient mutant of the Toxoplasma gondii ME49 strain and to test the virulence of the mutant. Methods Gene-deficient parasites were generated with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. Guide RNA (gRNA) was designed using the E-CRISPR software, and mutated on the pSAG1::Cas9-U6::sgUPRT plasmid using the Q5 site-directed mutagenesis kit to generate the pSAG1::Cas9-U6::sgGRA3 plasmid. A GRA3 donor plasmid containing GRA3 gene upstream sequences, pyrimethamine resistant gene dihydrofolate reductase-thymidylate synthase (DHFR-TS) and GRA3 gene downstream sequence was generated, and GRA3 donor DNA was amplified using PCR assay. The pSAG1::Cas9-U6::sgGRA3 plasmid and GRA3 donor DNA were electroporated into tachyzoites of the wild-type T. gondii ME49 strain. Then, parasite suspensions were inoculated into human foreskin fibroblast (HFF) cells and screened with pyrimethamine to yield pyrimethamine-resistant parasites for monoclonal screening. The GRA3 gene deficient monoclonal strain (ME49Δgra3) of T. gondii was identified using PCR and Western blotting assays, and the expression of GRA3 protein was determined in the T. gondii ME49Δgra3 strain using Western blotting. Subsequently, 1 000 freshly lysed tachyzoites of T. gondii ME49 and ME49Δgra3 strains were transferred to 12-well plates seeded with HFF cells, and incubated at 37 °C containing 5% CO₂ for 7 days, and the number of plaques was counted by staining with crystal violet solutions. HFF cells infected with tachyzoites of T. gondii ME49 and ME49Δgra3 strains were stained using Giemsa solutions, and the numbers of cells containing 1, 2, 4, and > 4 T. gondii parasitophorous vacuoles were counted. In addition, the survival rates of C57BL/6 mice infected with T. gondii ME49 and ME49Δgra3 strains were compared 35 days post-infection. Results PCR assay revealed successful amplification of both the upstream and downstream homologous arm bands of the DHFR-TS gene in the T. gondii ME49Δgra3 strain, and no corresponding bands were amplified in the ME49 strain. The GRA3 band was amplified in the ME49 strain, and the DHFR-TS band, rather than GRA3 band, was amplified in the ME49Δgra3 strain. Western blotting determined absence of GRA3 protein expression in the ME49Δgra3 strain. Crystal violet staining showed that the T. gondii ME49 strain produced more plaques than the ME49Δgra3 strain [(352.67 ± 26.39) plaques vs. (235.00 ± 26.29) plaques; t = 5.472, P < 0.01], and Giemsa staining revealed that the proportion of T. gondii parasitophorous vacuoles containing at least four T. gondii tachyzoites was higher in HFF cells infected with the ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(75.67 ± 2.52)% vs. (59.67 ± 2.31)%; t = 8.113, P < 0.01], and the proportion of T. gondii parasitophorous vacuoles containing at least 1 or 2 T. gondii tachyzoites was higher in HFF cells infected with the T. gondii ME49 strain than in those infected with the T. gondii ME49Δgra3 strain [(24.33 ± 2.52)% vs. (40.33 ± 2.31)%; t = −8.113, P < 0.01]. In addition, mice infected with the T. gondii ME49 and ME49Δgra3 strains started to die 8 and 9 days post-infection, and the 35-day mortality rates of mice infected with T. gondii ME49 and ME49Δgra3 strains were 10.00% and 70.00% post-infection (χ² = 6.762, P < 0.01). Conclusions The T. gondii ME49Δgra3 strain has been successfully generated, and GRA3 protein may increase the virulence of the T. gondii ME49 strain.

The rupture of carotid vulnerable plaques is the core pathological basis for major cardiovascular and cerebrovascular events. However， the insufficient alignment between existing animal models and the clinical disease and syndrome characteristics of traditional Chinese and western medicine has limited research progress. In this study， biomedical databases in China and abroad were systematically searched， and the modeling mechanisms and evaluation systems of carotid vulnerable plaque animal models were systematically assessed based on diagnostic criteria of both traditional Chinese and western medicine. Analysis of the clinical correspondence indicated that existing animal models can be categorized into four types： simple high-fat diet， surgical induction combined with high-fat feeding， genetic engineering combined with high-fat feeding， and drug induction combined with high-fat feeding. Among these， the compound strategy of surgical induction combined with high-fat feeding has become the current mainstream approach， showing good concordance with western medicine. The study found that the double balloon injury rabbit model and the ApoE^-/- mouse carotid artery tandem constriction combined with high-fat feeding model demonstrated a high degree of clinical correspondence with both traditional Chinese and western medicine in terms of vulnerable plaque imaging and pathological features. Nevertheless， existing models still face significant technical limitations in faithfully simulating plaque pathology and in translating findings to clinical applications. To address these challenges， integrating complex comorbidity mechanism construction， multimodal dynamic mechanism monitoring， and collaborative evaluation systems of traditional Chinese and western medicine could enable the development of highly concordant carotid vulnerable plaque disease-syndrome combination animal models. Such models would provide a reproducible experimental platform for targeted drug development to regulate plaque stability and for individualized precision treatment， as well as a theoretical basis for innovation in clinical diagnostic and therapeutic strategies.

With the growing popularity of video assisted thoracic surgery(VATS), the concept of enhanced recovery after surgery(ERAS) has become widely recognized. Alongside the emphasis on minimally invasive techniques during surgery, there is also a heightened focus on the management of postoperative thoracic drainage. The placement of a chest tube after thoracoscopic surgery often leads to wound pain, which can hinder patient recover. Numerous studies have demonstrated that for sublobar resections, lobectomies, and mediastinal procedures, it is safe and feasible to forgo chest tube placement, thereby accelerating patient recovery. This article reviews the structural anatomy of the thoracic cavity and the mechanisms underlying pleural effusion, drawing on pertinent research findings from both domestic and international studies. It examines the current practices regarding the postoperative placement of chest drainage tubes, evaluates the safety and feasibility of omitting such tubes, and outlines tube-free strategies for various surgical procedures. Additionally, the article addresses associated complications and their prevention.

Objective To explore the long-term and short-term therapeutic effects and mechanisms of electroacupuncture at Baihui(GV20)point and Dazhui(GV14)point in rats with ischemic stroke.Methods Thirty rats were randomly divided into a sham-operated group,a model group,and an electroacupuncture group,with 10 rats in each group.The model group and the electroacupuncture group were subjected to middle cerebral artery occlusion(MCAO)using the modified Longa method.The electroacupuncture group received electroacupuncture at Baihui point and Dazhui point once daily for one week.Long-term neurological recovery was assessed using the adhesive removal test and the forelimb placement test at 14,21 and 28 days after MCAO.On the second day after MCAO,cerebral cortical histopathological changes were observed using transmission electron microscopy,the expression of Iba-1 in cortical tissue was detected by immunofluorescence,and the expression levels of hypoxia-inducible factor 1α(HIF-1α),vascular endothelial growth factor(VEGF),interleukin 1β(IL-1β),and tumor necrosis factor α(TNF-α)in cortical tissue were measured by Western Blot.Results(1)Compared with the sham-operated group at the same time points,the model group showed prolonged adhesive removal time and increased forelimb stepping times at 14,21 and 28 days after MCAO(P＜0.05).Compared with the model group at the same time points,the electroacupuncture group exhibited significantly shortened adhesive removal time and reduced forelimb stepping times at 14,21 and 28 days after MCAO(P＜0.05).Within 28 days after MCAO,the adhesive removal time was gradually decreased,and forelimb stepping times were gradually reduced.(2)On the second day after MCAO:compared with the sham-operated group,the model group showed degeneration and necrosis of microglia in cortical ischemic penumbra,mitochondrial damage,increased expression of Iba-1 in cortical tissue,and elevated levels of HIF-1α,VEGF,IL-1β and TNF-α,with statistically significant differences(P＜0.05).Compared with the model group,the electroacupuncture group exhibited the improvement of microglia injury in cortical ischemic penumbra,reduced expression of Iba-1 in cortical tissue and decreased levels of HIF-1α,VEGF,IL-1β and TNF-α,with statistically significant differences(P＜0.05).Conclusion Electroacupuncture at Baihui point and Dazhui point can improve symptoms in rats with ischemic stroke in the long term and ameliorate degeneration and necrosis of cortical microglia in the ischemic penumbra in the short term.The mechanism may be related to the downregulation of HIF-1α,VEGF,IL-1β and TNF-α expressions in cerebral cortical tissue and the inhibition of microglial transformation to the M1 phenotype.

This study aims to elucidate the successful extubation process of a premature infant encountering difficulties due to an intracavicular knot at the proximal part of peripherally inserted central catheter (PICC). The reason for the difficulty in removing the catheter was determined by X-ray to be the knot at the proximal end of the PICC through the periphery. With the cooperation of a nurse specializing in static therapy and an interventional physician, the catheter was successfully removed based on the improved modified Sedinger technique .

This study aims to elucidate the successful extubation process of a premature infant encountering difficulties due to an intracavicular knot at the proximal part of peripherally inserted central catheter (PICC). The reason for the difficulty in removing the catheter was determined by X-ray to be the knot at the proximal end of the PICC through the periphery. With the cooperation of a nurse specializing in static therapy and an interventional physician, the catheter was successfully removed based on the improved modified Sedinger technique .

With the growing popularity of video assisted thoracic surgery(VATS), the concept of enhanced recovery after surgery(ERAS) has become widely recognized. Alongside the emphasis on minimally invasive techniques during surgery, there is also a heightened focus on the management of postoperative thoracic drainage. The placement of a chest tube after thoracoscopic surgery often leads to wound pain, which can hinder patient recover. Numerous studies have demonstrated that for sublobar resections, lobectomies, and mediastinal procedures, it is safe and feasible to forgo chest tube placement, thereby accelerating patient recovery. This article reviews the structural anatomy of the thoracic cavity and the mechanisms underlying pleural effusion, drawing on pertinent research findings from both domestic and international studies. It examines the current practices regarding the postoperative placement of chest drainage tubes, evaluates the safety and feasibility of omitting such tubes, and outlines tube-free strategies for various surgical procedures. Additionally, the article addresses associated complications and their prevention.

Objective:To compare the aneuploidy rate of embryos between poor ovarian response (POR) patients and women with normal ovarian reserve stratified by age, and to eliminate the influence of ovarian reserve on embryo quality.Methods:This was a retrospective case-control study of patients who underwent preimplantation genetic testing for aneuploidies (PGT-A) at the Department of Reproduction and Genetics in Hospital for Reproductive Medicine, Shandong University from January 2017 to December 2020. According to the POSEIDON criteria, POR patients were divided into group 1 [age<35 years, anti-Müllerian hormone (AMH)≥1.2 μg/L, number of oocytes retrieved≤9, 258 cycles], group 2 (age≥35 years, AMH≥1.2 μg/L, number of oocytes retrieved≤9, 397 cycles), group 3 (age<35 years, AMH<1.2 μg/L, number of oocytes retrieved≤9, 99 cycles) and group 4 (age≥35 years, AMH<1.2 μg/L, number of oocytes retrieved≤9, 377 cycles). The aneuploidy rate of the blastocysts in each group was compared with age-matched control women with normal ovarian reserve and normal ovarian response (non-POR 1 group, non-POR 2 group, non-POR 3 group and non-POR 4 group, AMH≥1.2 μg/L, number of oocytes retrieved>9). Based on the difference in sample size of POR groups and control groups, a 1∶2 propensity score matching (PSM) analysis was performed between the <35 years old POR groups and age-matched control groups (POR 1 group vs. non-POR 1 group, POR 3 group vs. non-POR 3 group) and a 1∶1 PSM analysis was performed between the ≥35 years old POR groups and age-matched control groups (POR 2 group vs. non-POR 2 group, POR 4 group vs. non-POR 4 group). The main outcomes were the rates of euploid and aneuploid embryo, the secondary outcomes were the numbers of oocytes retrieved, metaphaseⅡ oocytes, two pronuclei, embryos biopsied, euploid embryos, aneuploid embryos and mosaic embryos per cycle. Results:The number of oocytes retrieved and embryos biopsied embryos in POR 1-4 groups was significantly decreased compared with non-POR 1-4 groups [No. of oocytes retrieved: 7.0 (6.0, 9.0) vs. 16.0 (13.0, 20.0), 7.0 (5.0, 8.0) vs. 14.0 (11.0, 17.0), 6.0 (4.0, 9.0) vs. 16.0 (13.0, 20.0), 5.0 (3.0, 7.0) vs. 13.0 (11.0, 17.0), all P<0.001; No. of embryos biopsied: 3.0 (2.0, 4.0) vs. 4.0 (3.0, 6.0), 2.0 (1.0,3.0) vs. 3.0 (2.0, 5), 3.0 (2.0, 4.0) vs. 4.0 (3.0, 6.0), 2.0 (1.0, 3.0) vs. 3.0 (2.0, 5.0), all P<0.001]. After adjusting for repeated egg retrieval, PGT-A indications, ovarian stimulation protocol and total dosage of gonadotrophin, the embryo aneuploidy rate in group 4 POR patients was significantly higher than controls ( OR=1.252, 95% CI:1.053-1.488, P=0.011). However, no differences were identified in embryo aneuploidy rate between POR patients in groups 1, 2, 3 and corresponding controls, respectively (all P>0.05). Conclusion:The ovarian reserve adversely affects the quantity and quality of oocytes in advanced age POR women (≥35 years old). Decreased ovarian reserve in young women (<35 years old) mainly affects the number of oocytes retrieved.

Objective:To compare the aneuploidy rate of embryos between poor ovarian response (POR) patients and women with normal ovarian reserve stratified by age, and to eliminate the influence of ovarian reserve on embryo quality.Methods:This was a retrospective case-control study of patients who underwent preimplantation genetic testing for aneuploidies (PGT-A) at the Department of Reproduction and Genetics in Hospital for Reproductive Medicine, Shandong University from January 2017 to December 2020. According to the POSEIDON criteria, POR patients were divided into group 1 [age<35 years, anti-Müllerian hormone (AMH)≥1.2 μg/L, number of oocytes retrieved≤9, 258 cycles], group 2 (age≥35 years, AMH≥1.2 μg/L, number of oocytes retrieved≤9, 397 cycles), group 3 (age<35 years, AMH<1.2 μg/L, number of oocytes retrieved≤9, 99 cycles) and group 4 (age≥35 years, AMH<1.2 μg/L, number of oocytes retrieved≤9, 377 cycles). The aneuploidy rate of the blastocysts in each group was compared with age-matched control women with normal ovarian reserve and normal ovarian response (non-POR 1 group, non-POR 2 group, non-POR 3 group and non-POR 4 group, AMH≥1.2 μg/L, number of oocytes retrieved>9). Based on the difference in sample size of POR groups and control groups, a 1∶2 propensity score matching (PSM) analysis was performed between the <35 years old POR groups and age-matched control groups (POR 1 group vs. non-POR 1 group, POR 3 group vs. non-POR 3 group) and a 1∶1 PSM analysis was performed between the ≥35 years old POR groups and age-matched control groups (POR 2 group vs. non-POR 2 group, POR 4 group vs. non-POR 4 group). The main outcomes were the rates of euploid and aneuploid embryo, the secondary outcomes were the numbers of oocytes retrieved, metaphaseⅡ oocytes, two pronuclei, embryos biopsied, euploid embryos, aneuploid embryos and mosaic embryos per cycle. Results:The number of oocytes retrieved and embryos biopsied embryos in POR 1-4 groups was significantly decreased compared with non-POR 1-4 groups [No. of oocytes retrieved: 7.0 (6.0, 9.0) vs. 16.0 (13.0, 20.0), 7.0 (5.0, 8.0) vs. 14.0 (11.0, 17.0), 6.0 (4.0, 9.0) vs. 16.0 (13.0, 20.0), 5.0 (3.0, 7.0) vs. 13.0 (11.0, 17.0), all P<0.001; No. of embryos biopsied: 3.0 (2.0, 4.0) vs. 4.0 (3.0, 6.0), 2.0 (1.0,3.0) vs. 3.0 (2.0, 5), 3.0 (2.0, 4.0) vs. 4.0 (3.0, 6.0), 2.0 (1.0, 3.0) vs. 3.0 (2.0, 5.0), all P<0.001]. After adjusting for repeated egg retrieval, PGT-A indications, ovarian stimulation protocol and total dosage of gonadotrophin, the embryo aneuploidy rate in group 4 POR patients was significantly higher than controls ( OR=1.252, 95% CI:1.053-1.488, P=0.011). However, no differences were identified in embryo aneuploidy rate between POR patients in groups 1, 2, 3 and corresponding controls, respectively (all P>0.05). Conclusion:The ovarian reserve adversely affects the quantity and quality of oocytes in advanced age POR women (≥35 years old). Decreased ovarian reserve in young women (<35 years old) mainly affects the number of oocytes retrieved.