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 provide valuable insights for the adjustment of anti-infectious regimens， identification of adverse reactions， and individualized pharmaceutical care in patients with critically severe scrub typhus. METHODS Clinical pharmacists actively participated in the pharmaceutical care process for a patient with severe scrub typhus leading to mixed shock undergoing continuous renal replacement therapy and extracorporeal membrane oxygenation. Initially， the patient received meropenem （1 g， q12 h， ivdrip）， in combination with doxycycline （0.1 g， q12 h， po）， which was later switched to meropenem （1 g， q8 h， ivdrip） along with omacycline （100 mg， qd， ivdrip） due to impaired gastrointestinal function. However， as the patient’s condition progressively deteriorated and the infection became uncontrolled， the clinical pharmacists recommended that the clinicians adjust the anti-infective regimen to meropenem （2 g， q8 h， ivdrip） combined with tigecycline （100 mg for first dose； 50 mg， q12 h for maintenance； ivdrip）. The clinicians followed the advice of the clinical pharmacists. After treatment， the patient’s symptoms exhibited significant improvement， accompanied by a notable decrease in inflammatory markers， indicating that the infection had been successfully controlled. However， due to continuously increasing bilirubin levels， in order to reduce the risk of drug-induced liver injury， the clinicians changed tigecycline to azithromycin （0.5 g， qd， ivdrip） following the recommendation of the clinical pharmacists. RESULTS Ultimately， metagenomic next-generation sequencing of the bronchoalveolar lavage fluid and blood specimens indicated that Orientia tsutsugamushi had been completely eradicated in the patient. CONCLUSIONS Tigecycline may be a viable therapeutic choice for patients with severe scrub typhus. In the context of critically ill patients with scrub typhus， combining tigecycline with azithromycin might potentially enhance the efficacy in eliminating Orientia tsutsugamushi.

BACKGROUND:Previous studies have shown that gelatin modified by methacrylic anhydride(GelMA)/treated dentin matrix(TDM)composite hydrogel scaffolds can promote the proliferation and differentiation of human dental pulp stem cells.Hydrogels loaded with bone morphogenetic protein 2 are considered to be promising materials for bone repair.OBJECTIVE:To observe the effect of GelMA/TDM composite hydrogels loaded with different concentrations of bone morphogenetic protein-2 on the induction of osteogenic differentiation of human dental pulp stem cells.METHODS:GelMA/TDM composite hydrogels containing 0,50,100,and 200 μg/mL bone morphogenetic protein 2 were prepared,denoted as GelMA/TDM,bone morphogenetic protein-2(50 ng/mL)GelMA/TDM,bone morphogenetic protein-2(100 ng/mL)GelMA/TDM,and bone morphogenetic protein-2(200 ng/mL)GelMA/TDM,respectively,and the in vitro sustained release of bone morphogenetic protein 2 from the composite hydrogels was detected.Human dental pulp stem cells were extracted by the modified tissue block enzymatic digestion method and seeded on the surfaces of the four kinds of hydrogels.Cell proliferation was detected by CCK-8 assay and cell adhesion was detected by DAPI staining.Human dental pulp stem cells on the surfaces of each group of hydrogels were induced to osteoblastically,and alkaline phosphatase staining,alkaline phosphatase activity detection,and alizarin red staining were performed.RT-PCR was used to detect the expression of related osteogenic genes(Runx2,bone morphogenetic protein 2,osteopontin,osteocalcin,and type Ⅰ collagen).RESULTS AND CONCLUSION:(1)The GelMA/TDM composite hydrogel could continuously release bone morphogenetic protein 2 for up to 21 days,with a faster release from day 3 to day 6 and a steady release after 6 days.(2)All four kinds of hydrogels could promote the proliferation of human dental pulp stem cells,among which the bone morphogenetic protein-2(100 ng/mL)-GelMA/TDM composite hydrogel had the most obvious effect.Compared with the GelMA/TDM composite hydrogel,the bone morphogenetic protein-2-GelMA/TDM composite hydrogel could promote the adhesion of human dental pulp stem cells,among which the bone morphogenetic protein-2(200 ng/mL)-GelMA/TDM composite hydrogel had the most obvious effect.(3)Compared with the GelMA/TDM composite hydrogel,the bone morphogenetic protein-2-GelMA/TDM composite hydrogel could increase the activity of alkaline phosphatase,the content of calcium nodules,and the expression of related osteogenic genes.Comprehensive analysis showed that the effect of the bone morphogenetic protein-2(100 ng/mL)-GelMA/TDM composite hydrogel was more obvious.(4)The results show that the bone morphogenetic protein-2(100 ng/mL)-GelMA/TDM composite hydrogel has a more obvious ability to promote the osteogenic differentiation of dental pulp stem cells.

Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25％hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25％HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90％,specificity of 100％,positive prediction rate of 100％,negative prediction rate of 83％,false positive rate of 0％,false negative rate of 17％and accuracy of 93％.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.

Objective To develop a predictive model for postoperative mortality risk in patients with acute aortic dissection(AAD)using the Extreme Gradient Boosting(XGBoost)algorithm combined with Shapley Additive Explanation(SHAP),and to establish a prediction website to serve as a diagnostic and therapeutic support platform for clinicians and patients.Methods A retrospective cohort study design was adopted.Data from 782 AAD patients who underwent surgical treatment at the Fourth Hospital of Hebei Medical University from January 2013 to December 2023 were collected,including basic information and initial serum biomarker test results.Patients were randomly divided into training and test sets at a 7:3 ratio.An external validation set consisting of 313 AAD patients admitted to the Second Hospital of Hebei Medical University from January 2020 to December 2023 was also established for further model validation.Variables were screened using LASSO regression,and an XGBoost machine learning model was constructed and interpreted using SHAP.The predictive performance of the model was evaluated using receiver operating characteristic(ROC)curve analysis.Using the Shiny package,the XGBoost model was deployed to shinyapps.io to create a prediction website for postoperative mortality risk in AAD patients.One patient was selected by simple random sampling from the test set and the external validation set respectively for the prediction example on the Shiny webpage.Results The XGBoost model demonstrated high predictive performance for postoperative mortality in AAD patients,with area under the ROC curve(AUC)values of 0.928(95%CI 0.901-0.956)in the training set,0.919(95%CI 0.891-0.949)in the test set,and 0.941(95%CI 0.915-0.967)in the external validation set.SHAP values indicated the following order of variable importance in the model(from highest to lowest):"lactate dehydrogenase""blood chlorine""multiple organ injury""carbon dioxide combining power""prothrombin time""α-hydroxybutyric acid""creatine kinase isoenzyme""Stanford classification""combined use of bedside blood purification""gender""acute kidney injury""gastrointestinal bleeding""brain injury"and"shock".A risk prediction website for adverse postoperative outcomes in AAD patients was developed using XGBoost-SHAP method(https://dun-dunxiaolu.shinyapps.io/document/)and validated with examples.One randomly selected patient from each of the test and external validation sets was applied:the predicted mortality risk value for patient 1(who died postoperatively)was 0.9539,and that for patient 2(who survived postoperatively)was 0.0206.Conclusions The XGBoost-SHAP model demonstrates high accuracy in predicting postoperative mortality risk for AAD patients.The online prediction tool established based on this model enhances the identification efficiency of high-risk postoperative mortality patients.

Objective:To analyze policy differences in general outpatient fund pooling of employee basic medical insurance in Yangtze River Delta Region and raise suggestions from the perspective of coordinated development.Methods:The study analyzed policy documents on general outpatients financing,outpatient chronic and special diseases,and inclusion of retail pharmacies into risk pooling at provincial level and in some economically developed cities before August,2024.Typical cases were selected for detailed introduction.Results:General outpatient coverage of employee medical insurance in Yangtze River Delta Region is high,with individual account payment scenarios expanding to commercial health insurance,Long Term Care Insurance and family physician contracted service fees.Number of outpatient chronic and special diseases ranges from 3 to 83.Retail pharmacy policies differ less in access conditions and treatment and e-prescription is advancing rapidly.Suzhou is actively exploring total budget management of retail pharmacies.Conclusions:Outpatient fund pooling policies should be gradually promoted under the principle of"differentiated coordination".It is necessary to transform outpatient chronic and special diseases from"disease coverage"to"cost coverage";strengthen the management of retail pharmacies under the outpatient fund pooling scheme,and improve the coordinated regulatory mechanism for direct settlement of outpatient and medication expenses incurred in other places in the Yangtze River Delta.

Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25％hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25％HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90％,specificity of 100％,positive prediction rate of 100％,negative prediction rate of 83％,false positive rate of 0％,false negative rate of 17％and accuracy of 93％.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.

Objective To evaluate the cost-effectiveness of the combination therapy of ribociclib plus fulvestrant compared to fulvestrant monotherapy as a second-line treatment for postmenopausal patients with hormone receptor-positive(HR+)/human epidermal growth factor receptor 2-negative(HER2-)advanced metastatic breast cancer from the perspective of the Chinese healthcare system.Methods A partitioned survival model was constructed based on MONALEESA-3 study data,with the model cycle of 4 weeks and the time range of 15 years.Total cost,quality-adjusted life years(QALYs)and incremental cost-effectiveness ratio(ICER)were compared between ribociclib combined with fulvestrant and fulvestrant monotherapy as a second-line treatment for postmenopausal patients with HR+/HER2-advanced metastatic breast cancer.The robustness of the models was validated through one-way sensitivity analysis and probabilistic sensitivity analysis.Results Compared with the fulvestrant monotherapy regimen,the ICER for the ribociclib combined with fulvestrant regimen was 187,958.06 yuan/QALY,which was below the willingness-to-pay(WTP)threshold of three times China's per capita gross domestic product(GDP)in 2024(287,247 yuan/QALY).The one-way sensitivity analysis revealed that the ICER was primarily influenced by the proportion of patients receiving subsequent treatments with the combination therapy or fulvestrant monotherapy,and the utility value of progression-free survival.The probabilistic sensitivity analysis showed that at a WTP threshold of three times China's per capita GDP in China in 2024(287,247 yuan/QALY),the probability of the ribociclib combination therapy being cost-effective was 100%.Conclusion In the context of the Chinese healthcare system,the combination therapy of ribociclib and fulvestrant is more cost-effective than fulvestrant monotherapy for the second-line treatment of postmenopausal patients with HR+/HER2-advanced metastatic breast cancer.

A primary hallmark of pathological cardiac hypertrophy is excess protein synthesis due to enhanced translational activity. However, regulatory mechanisms at the translational level under cardiac stress remain poorly understood. Here we examined the translational regulations in a mouse cardiac hypertrophy model induced by transaortic constriction (TAC) and explored the conservative networks versus the translatome pattern in human dilated cardiomyopathy (DCM). The results showed that the heart weight to body weight ratio was significantly elevated, and the ejection fraction and fractional shortening significantly decreased 8 weeks after TAC. Puromycin incorporation assay showed that TAC significantly increased protein synthesis rate in the left ventricle. RNA-seq revealed 1,632 differentially expressed genes showing functional enrichment in pathways including extracellular matrix remodeling, metabolic processes, and signaling cascades associated with pathological cardiomyocyte growth. When combined with ribosome profiling analysis, we revealed that translation efficiency (TE) of 1,495 genes was enhanced, while the TE of 933 genes was inhibited following TAC. In DCM patients, 1,354 genes were upregulated versus 1,213 genes were downregulated at the translation level. Although the majority of the genes were not shared between mouse and human, we identified 93 genes, including Nos3, Kcnj8, Adcy4, Itpr1, Fasn, Scd1, etc., with highly conserved translational regulations. These genes were remarkably associated with myocardial function, signal transduction, and energy metabolism, particularly related to cGMP-PKG signaling and fatty acid metabolism. Motif analysis revealed enriched regulatory elements in the 5' untranslated regions (5'UTRs) of transcripts with differential TE, which exhibited strong cross-species sequence conservation. Our study revealed novel regulatory mechanisms at the translational level in cardiac hypertrophy and identified conserved translation-sensitive targets with potential applications to treat cardiac hypertrophy and heart failure in the clinic.
Animals ; Humans ; Cardiomegaly/physiopathology* ; Ribosomes/physiology* ; Protein Biosynthesis/physiology* ; Mice ; Cardiomyopathy, Dilated/genetics* ; Ribosome Profiling