Objective To explore the technical process and clinical application of laparoscopic combined upper midline incision minimally invasive liver transplantation. Methods A retrospective analysis was conducted on 30 cases of laparoscopic combined upper midline incision minimally invasive liver transplantation. The cases were divided into cirrhosis group (15 cases) and liver failure group (15 cases) based on the primary disease. The surgical and postoperative conditions of the two groups were compared. Results All patients successfully underwent laparoscopic "clockwise" liver resection, with no cases of passive conversion to open surgery or intolerance to pneumoperitoneum. In 6 cases, the right lobe was relatively large, and the right hepatic ligaments could not be completely mobilized. One case required an additional reverse "L" incision during open surgery. All patients successfully completed the liver transplantation, with no major intraoperative bleeding, cardiovascular events, or other occurrences in the 30 patients. The model for end-stage liver disease (MELD) score in the cirrhosis group was lower than that in the liver failure group (P<0.001). There were no statistically significant differences between the two groups in terms of age, surgical time, blood loss, anhepatic phase, or cold ischemia time (all P>0.05). During the perioperative period, there was 1 case of hepatic artery embolism, 1 case of portal vein anastomotic stenosis, no complications of hepatic vein and inferior vena cava, and 3 cases of biliary anastomotic stenosis, all of which occurred in the liver failure group. Conclusions In strictly selected cases, the minimally invasive liver transplantation technique combining laparoscopic hepatectomy with upper midline incision for graft implantation has the advantages of smaller incisions, less bleeding, relatively easier operation, and faster postoperative recovery, which is worthy of clinical promotion and application.

Objective: To explore the active components, key targets, and mechanism of action of turnip in alleviating pulmonary fibrosis(PF) based on network pharmacology and animal experiments. Methods: The active components and targets of Brassica rapa L. were screened using the traditional Chinese medicine systems pharmacology database and analysis platform database, and PF-related targets were obtained from disease databases such as online mendelian inheritance of man(OMIM) and DrugBank. The intersection targets were used to construct a protein-protein interaction(PPI) network to identify core targets, followed by gene oncology(GO)/Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis. In the animal experiments, a bleomycin-induced PF mouse model was established. Pathological changes in lung tissue were evaluated using HE and Masson staining. qRT-PCR was used to detect the mRNA expression of tumor necrosis factor-α(TNF-α), phosphatidylinositol 3-kinase(PI3K), and akstrain transforming 1(AKT1), and immunofluorescence staining was used to measure the protein expression of TNF-α, PI3K, and AKT1. Results: The 68 active components identified in Brassica rapa L. may regulate PI3K-Akt signaling pathway by acting on 89 potential targets such as TNF-α and AKT1. The results of animal experiments showed that polysaccharide of Brassica rapa L.(BRPs) could significantly reduce the degree of bleomycin induced pulmonary fibrosis in mice; HE and Masson staining of lung tissue showed that compared with the model group, the damage of alveolar structure, the infiltration of inflammatory cells and the deposition of collagen fibers in the BRPs treatment group were significantly reduced. Further mechanism studies showed that BRPs could significantly down-regulate the mRNA and protein expression levels of TNF-α, PI3K and AKT1 in lung tissue of pulmonary fibrosis mice. Conclusion Brassica rapa L. can synergistically alleviate pulmonary fibrosis through “multi-component, multi-target and multi-channel” approach; BRPs is one of the main active components, and plays an anti-fibrosis role by inhibiting TNF-α/PI3K Akt signaling pathway.

Objective To investigate immunogenic and toxic effects of graphene oxide (GO) nanoparticles in mouse skeletal muscles and in human blood in vitro. Methods GO nanoparticles prepared using a probe sonicator were supended in deionized H2O or PBS, and particle size and surface charge of the nanoparticles were measured with dynamic light scattering (DLS). Different concentrations (0.5, 1.0 and 2.0 mg/mL) of GO suspension or PBS were injected at multiple sites in the gastrocnemius muscle (GN) of C57BL/6 mice, and inflammatory response and immune cell infiltrations were detected with HE and immunofluorescence staining. We also examined the effects of GO nanoparticles on human red blood cell (RBC) morphology, hemolysis and blood coagulation using scanning electron microscope (SEM), spectrophotometry, and thromboelastography (TEG). Results GO nanoparticles suspended in PBS exhibited better colloidal dispersity, stability and surface charge effects than those in deionized H2O. In mouse GNs, injection of GO suspensions dose- and time-dependently resulted in sustained muscular inflammation and myofiber degeneration at the injection sites, which lasted till 8 weeks after the injection; immunofluorescence staining revealed obvious infiltration of monocytes, macrophages, dendritic cells and CD4+T cells around the injection sites in mouse GNs. In human RBCs, incubation with GO suspensions at 0.2, 2.0 and 20 mg/mL, but not at 0.002 or 0.02 mg/mL, caused significant alterations of cell morphology and hemolysis. TEG analysis showed significant abnormalities of blood coagulation parameters following treatment with high concentrations of GO. Conclusion GO nanoparticles can induce sustained inflammatory and immunological responses in mouse GNs and cause RBC hemolysis and blood coagulation impairment, suggesting its muscular toxicity and hematotoxicity at high concentrations.

Objective To investigate immunogenic and toxic effects of graphene oxide (GO) nanoparticles in mouse skeletal muscles and in human blood in vitro. Methods GO nanoparticles prepared using a probe sonicator were supended in deionized H2O or PBS, and particle size and surface charge of the nanoparticles were measured with dynamic light scattering (DLS). Different concentrations (0.5, 1.0 and 2.0 mg/mL) of GO suspension or PBS were injected at multiple sites in the gastrocnemius muscle (GN) of C57BL/6 mice, and inflammatory response and immune cell infiltrations were detected with HE and immunofluorescence staining. We also examined the effects of GO nanoparticles on human red blood cell (RBC) morphology, hemolysis and blood coagulation using scanning electron microscope (SEM), spectrophotometry, and thromboelastography (TEG). Results GO nanoparticles suspended in PBS exhibited better colloidal dispersity, stability and surface charge effects than those in deionized H2O. In mouse GNs, injection of GO suspensions dose- and time-dependently resulted in sustained muscular inflammation and myofiber degeneration at the injection sites, which lasted till 8 weeks after the injection; immunofluorescence staining revealed obvious infiltration of monocytes, macrophages, dendritic cells and CD4+T cells around the injection sites in mouse GNs. In human RBCs, incubation with GO suspensions at 0.2, 2.0 and 20 mg/mL, but not at 0.002 or 0.02 mg/mL, caused significant alterations of cell morphology and hemolysis. TEG analysis showed significant abnormalities of blood coagulation parameters following treatment with high concentrations of GO. Conclusion GO nanoparticles can induce sustained inflammatory and immunological responses in mouse GNs and cause RBC hemolysis and blood coagulation impairment, suggesting its muscular toxicity and hematotoxicity at high concentrations.

Aim To construct a diabetic atherosclerosis mouse model and study the pathological characteristics of diabetic atherosclerosis.Methods Fifty 8-week-old male LDLR-/-mice were fed with standard diet for 2 weeks and then changed to high-fat diet,they were randomly divided into two groups.The diabetic atherosclerosis group was given intraperitoneal injection of low dose streptozotocin(STZ)for 5 days continuouly to establish the model,and the atheroscle-rosis group was given citrate buffer injection at the same time.The body mass,blood glucose and blood lipids of the mice in the two groups were detected for many times.At the age of 23 weeks,the mice were euthanized after glucose tolerance test.HE staining and oil red O staining were used to detect the gross and aortic root atherosclerosis,immunohistochemical staining was used to detect CD4,α-smooth muscle actin(α-SMA),EGF-like module-containing mucin-like hormone re-ceptor-like 1(EMR1),monocyte chemotactic protein-1(MCP-1),NOD-like receptor protein 3(NLRP3),vascular cell adhesion molecule-1(VCAM-1),matrix metalloproteinase-2(MMP-2)and tissue inhibitor of metalloproteinase-1(TIMP-1),Western blot was used to detect α-SMA,CD4,tumor necrosis factor-α(TNF-α),NLPR3,intercellular adhesion molecule-1(ICAM-1),and type Ⅰ and Ⅲ collagen.Results Compared with the atherosclerosis group,the body mass decreased,the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDLC)increased,and the levels of high density lipoprotein cholesterol(HDLC)decreased(P＜0.05)in the diabetic atherosclerosis group.Compared with the atherosclerosis group,the distribution of atherosclerotic plaques was diffuse and the area was increased in the diabetic atherosclerosis group,and the contents of lipids,T cells,macrophages,smooth muscle cells,type Ⅰ and Ⅲ colla-gen were increased(P＜0.05);the protein levels of TNF-α,MCP-1,MMP-2,NLRP3,ICAM-1 and VCAM-1 in vascular tissues were increased,while the content of TIMP-1 were decreased and MMP2/TIMP-1 were increased(P＜0.05).Conclusions LDLR-mouse model of diabetic atherosclerosis can be successfully established by STZ induction combined with high-fat diet,which can reflect the plaque composition and inflammatory characteristics of diabetes promoting atheroscle-rosis.It can be used as a relatively ideal pathological model for the study of diabetic macroangiopathy.

OBJECTIVE To investigate the effects of 3 different primitives or the same primitives with different characters of Tibetan medicine Zuomoxing[Caragana changduenais Liou f. with red heartwood, Caragana jubata(Pall.) Poir. with brown and white heartwood] on rats with pattern of toxic heat-induced blood stasis. METHODS　Ninety SD rats were randomly divided into the blank group, model group, aspirin-positive group, Changdu low-dose group(CDD), Changdu high-dose group(CDG), whitewood of Guijian low-dose group(GJBD), whitewood of Guijian high-dose group(GJBG), brown wood of Guijian low-dose group(GJZD), Brown wood of Guijian high-dose group(GJZG). Models with heat toxicity and blood stasis pattern were established by intraabdominal injection of carrageenan combined with tail vein injection of lipopolysaccharide. The effects of each group on blood rheology, coagulation four indices and blood routine were determined, and the content of arachidonic acid(AA), IL-1β, IL-6, TNF-α and thromboxane B2(TXB2) were measured with ELISA. RESULTS ①Blood rheology: Compared with model group, CDD and CDG significantly decreased whole blood viscosity(WBV), reduction viscosity of whole blood(WBRV), erythrocyte rigidity index(HGX), erythrocyte deformability index(EDI), whole blood relative index(WBRI) (P<0.01), and increased plasma viscosity(P<0.01). GJZG and GJZD significantly decreased HGX(P<0.01 or P<0.05), and increased plasma viscosity(P<0.01). GJBG and GJBG significantly decreased WBHSV, WBHSRV, HGX, EDI, and whole blood high shear relative index(WBHSRI)(P<0.01). ②Coagulation four indices: Compared with model group, CDD significantly reduced the thrombin time(TT)(P<0.01). GJZG significantly reduced activated partial thromboplastin time(APTT) and TT(P<0.01 or P<0.05). GJBD significantly reduced prothrombin time(PT) and APTT (P<0.01 or P<0.05). ③Blood routine: Compared with model group, GJZD and GJBD significantly decreased the percentage of monocytes(P<0.01 or P<0.05). The number of large platelets in CDD significantly increased(P<0.05). CDG significantly increased the platelet number, platelet hematocrit, and large platelet number(P<0.01 or P<0.05), and tended which to be normal. ④Inflammatory factors: Compared with model group, the levels of TNF-α, IL-6, TXB2 were significantly increased in CDD and CDG(P<0.01 or P<0.05). The levels of IL-6 and TXB2 were significantly increased in GJZD and GJZG(P<0.01). GJBD was significantly increased TXB2(P<0.01), and GJBG was significantly increased IL-1β, IL-6, and TXB2(P<0.01), while decreased AA(P<0.05). CONCLUSION Zuomoxing with separate sources have different degrees of effects on rats with pattern of toxic heat-induced blood stasis, and have different degrees of effects on hemorheology, coagulation factors, blood routine and inflammatory mediators, and the degree and trend of effects are different with different doses. The effect of promoting blood circulation and removing blood stasis was generally manifested as Changdu > whitewood of Guijian > Brown wood of Guijian. The effect of promoting blood circulation and removing blood stasis may be the result of multiple pathways and mechanisms.

【Objective】 To investigate the factors influencing the length of hospital stays of the acute fatty liver of pregnancy (AFLP) patients, so as to establish the prediction model. 【Methods】 A total of 49 patients diagnosed as AFLP)in ShenZhen People’s Hospital between January 2008 and January 2023 were retrospectively analyzed. According to the median length of hospital stays, the patients were divided into two groups: Group A(n=21)and Group B(n=28). Preoperative general laboratory data, clinical features and postpartum adverse outcomes in both groups were analyzed. Multivariate binary logistic regression was used to analyze the independent factors affecting the length of hospital stays for AFLP, and a prediction model for hospitalization time was established. 【Results】 Comparison between Group B and Group A were as follows: hospital stays(d)(15.5 vs 8), preoperative icterus(%)[16(57.1%)vs 3(14.3%)], thrombin time(TT)(s)(24.2 vs 21.3), prothrombin time(PT)(s)(16.8 vs 15.3), activated partial thromboplastin time(APTT)(s)(52.3 vs 40.7), total bilirubin(TBIL)(μmol/L)(77.2 vs 45.2), indirect bilirubin(IBIL)(μmol/L)(21.2 vs 10), creatinine(Cre)(μmol/L)[(171.97±53.34) vs (131.81±45.06]), TT extension(%)[24(85.7%)vs 11(52.4%)], APTT extension(%)[27(96.4%)vs 7(33.3%)], IBIL elevation(%)[19(67.9%)vs 4(19%)], Cre concentration rise(%)[21(75%)vs 8(38%)], number of postpartum plasma exchange sessions(%)[23(82.1%)vs 5(23.8%)], postpregnancy co-infection phenomenon(%)[21(75%)vs 4(19%)], with Group B significantly higher than Group A. The preoperative platelet count(×10⁹/L)(128 vs 221)and the concentration of fibrinogen(g/L)[0.9 vs 1.6] in Group B were significantly lower than those in Group A. Univariate logistic regression analysis showed that preoperative icterus, postpregnancy co-infection phenomenon, number of postpartum plasma exchange sessions, preoperative TT extension, preoperative APTT extension, Cre concentration rise were influencing factors for the hospital stays of AFLP patients. According to the minimum result of Akaike information criterion, the multivariate binary logistic regression analysis (step-wise selection) showed that the number of postpartum plasma exchange sessions, icterus, preoperative APTT extension were the independent risk factor influencing the hospital stays of AFLP patients, and the logistic regression prediction model was established by incorporating the above three factors. Regularization techniques were further employed in linear regression to address and assess overfitting issues. Additionally, the confidence interval for the estimated effect sizes in each model have been acquired by bootstrapping techniques. 【Conclusion】 Preoperative icterus, preoperative APTT extension(APTT>43s)and the number of postpartum plasma exchange sessions were the independent risk factor influencing the hospital stays of AFLP patients and the logistic regression prediction model with high predictive effectiveness was established successfully.

ObjectiveTo study the correlation between the content of active ingredients of Aurantii Fructus in different main production areas and soil factors， so as to provide a theoretical basis for implementing ecological regulation of soil， improving the quality of Aurantii Fructus， and revealing the origin of genuine medicinal materials. MethodThe content of naringin， neohesperidin， total flavonoids， volatile oil， total nitrogen， total phosphorus， total potassium， and 17 soil factor-related indicators in 25 batches of Aurantii Fructus from different production areas were determined. The main soil factors affecting the content of active ingredients of Aurantii Fructus were analyzed by Pearson correlation analysis， principal component analysis， and grey correlation analysis. ResultThe pH value of the soil is between 4.83 and 8.21， and the soil is weakly acidic and neutral in general. Soil fertility exceeds the average. Pearson correlation analysis shows that the soil factors most related to the four active ingredients of Aurantii Fructus are total phosphorus， available copper， available zinc， exchangeable magnesium， available sulfur， available phosphorus， and available molybdenum. Principal component analysis shows that total nitrogen， alkali-hydrolyzable nitrogen， organic matter， available phosphorus， and available zinc are the main characteristic factors in soil. Grey correlation analysis shows that the main soil factors affecting the active ingredients of Aurantii Fructus are total phosphorus， total nitrogen， available zinc， available copper， exchangeable magnesium， and pH. ConclusionIn the cultivation of Aurantii Fructus， the medicinal material quality of Aurantii Fructus could be improved by adjusting the level of beneficial factors in the soil and improving the soil texture.

[Objective]This study aimed to improve the existing semen processing methods in the field of reproductive male medicine,particularly focusing on the 300×g 20 min treatment condition in the double-layer density gradient method,to enhance fertilization outcome.[Methods]Semen specimens from 1623 patients undergoing assisted reproductive techniques at the Reproductive Medicine Center of the Sixth Affiliated Hospital of Sun Yat-sen University from July and September 2020 and March and May 2022 were collected for preliminary experiments.Four different double-layer density gradient methods(200×g 10 min,200×g 20 min,300×g 10 min,and 300×g 20 min)were compared for sperm DNA fragmentation rates and recovery rates after processing.Subsequently,the optimal method was selected as the new approach and compared with the current method in use(300×g 20 min double-layer gradient method)to assess any statistical differences in fertilization rates.Further optimization to a single-layer density gradient method was performed based on the new method and compared with the double-layer density gradient method to determine any statistical differences.Experimental conditions were strictly controlled for temperature,centrifugation speed,and duration,with the quantity and processing conditions of each sample recorded.[Results]Among the four double-layer density gradient methods,the sperm DNA fragmentation rate was lower with the 300×g 10 min treatment compared to 300×g 20 min while ensuring sufficient sperm recovery rates.Consequently,the 300×g 10 min method was selected as the new approach for experimentation.Results indicated that the total fertilization rate and 2 pronuclei(2PN)fertilization rate with the new 300×g 10 min method were higher than with the 300×g 20 min method,the difference was statistically significant(P<0.05).Although the cleavage rate with 300×g 10 min was slightly higher than 300×g 20 min,the difference was not statistically significant(P>0.05).The total fertilization rate and 2PN fertilization rate were slightly higher with the single-layer density gradient method compared to the double-layer density gradient method,but the difference was not statistically significant(P>0.05).The cleavage rate with the single-layer density gradient method was higher than the double-layer density gradient method,and the blastocyst formation rate is lower than that of the double-layer density gradient method,and the differences are statistically significant(P<0.05).[Conclusion]The 300×g 10 min double-layer density gradient method successfully improved total fertilization rates,2PN fertilization rates,and cleavage rates compared to the existing 300×g 20 min method,while reducing the time required for semen optimization processing.Although the single-layer density gradient method improves the cleavage rate,and saves reagent costs and operation time,its blastocyst formation rate has decreased.These findings provide valuable guidance and insights for semen processing methods in the field of reproductive andrology.

Objective:To study the role of myocardial work parameters in early identification of myocardial injury in neonatal asphyxia.Methods:From July 2020 to December 2021, neonates diagnosed with mild neonatal asphyxia admitted to the Department of Neonatology of our hospital within 24 h after birth were prospectively enrolled into the asphyxia group. Neonates without asphyxia during the same period were selected as the control group and matched with the asphyxia group for gender, gestational age and birth weight at a ratio of 1:1~1:2. The asphyxia group was subgrouped into preterm asphyxia group and term asphyxia group. All neonates received echocardiography within 24 h after birth. Multiple parameters were measured including M-mode, two-dimensional image, Doppler image, global longitudinal strain (GLS) and myocardial work parameters [global work index (GWI), global constructive work (GCW), global wasted work (GWW), global work efficiency (GWE)]. The level of serum N-terminal pro brain natriuretic peptide (NT-proBNP) was recorded in the asphyxia group. The data were compared between the asphyxia group and the control group. Correlations between myocardial work parameters and other parameters were analyzed.Results:A total of 33 cases were in the asphyxia group and 43 cases were in the control group. The preterm asphyxia group (18 cases) showed significantly lower GWI and GCW than the preterm control group (18 cases) [GWI: (702±153) mmHg vs. (879±205) mmHg, GCW: (1 016±221) mmHg vs. (1 200±271) mmHg] ( P<0.05). No differences existed in GLS, GWW and GWE. The term asphyxia group (15 cases) showed significantly lower GWW than the term control group (25 cases) [45.0 (30.0, 65.0) mmHg vs. 71.0 (35.5,85.5) mmHg] ( P<0.05). No differences existed in GLS, GWI, GCW and GWE. GWI was negatively correlated with serum NT-proBNP level ( r=-0.327, P<0.05). Conclusions:GWI and GCW may indicate myocardial injury in preterm neonates with mild asphyxia.