Objective To develop an ion-pairing reversed-phase high performance liquid chromatography(IP-RP-HPLC)method for the determination of L-methionine sulphoximine(MSX) residues in recombinant protein therapeutics, and to verify and preliminarily apply it, so as to provide a reference for the quality control of related products.Methods A chromatographic analysis method was screened and the mobile phase conditions such as ion-pairing reagent, pH and salt concentration were optimized to develop an IP-RP-HPLC method for the determination of MSX residues in recombinant protein therapeutics. The method was verified for specificity, system applicability, linearity, limit of detection(LOD) and limit of quantitation(LOQ), precision, accuracy and solution stability, by which the residual MSX contents in multiple batches of in-process sample and bulk of recombinant protein therapeutics were determined.Results The chromatographic conditions were determined as follows: using Hypersil GOLDTMC18 column(4. 6 mm × 250 mm, 5 μm) at column temperature of 25 ℃; 50 mmol/L sodium dihydrogen phosphate(containing 5 mmol/L sodium 1-octane sulfonate, pH 2. 0)-acetonitrile(volume fraction of 85 ∶ 15) as mobile phase for isocratic elution at a flow rate of 0. 6 mL/min; detection wavelength of 200 nm; injection volume of 50 μL. The method exhibited good specificity and system applicability. The control substance showed good linearity in the concentration range of 10-100 μg/mL, with a correlation coefficient(R) of 0. 999 95. The LOD and LOQ were 5 and 0. 5 μg/mL, respectively. MSX was not detected in either recombinant protein A or B, and the relative standard deviations(RSDs) of the measured concentrations of six parallel spiked samples were 1. 3% and 1. 1%, respectively, with the recovery rates ranged from 94% to 103%. For recombinant protein A spiked test sample placed at 10 ℃, the RSD of the measured concentrations at 0 h and 12 h were 2. 1%. MSX was not detected in the in-process samples and bulk of recombinant protein A test samples and three batches of recombinant protein B samples.Conclusion The developed IP-RP-HPLC method has the characteristics of simple operation and high sensitivity, and can be used for the quality control of MSX residues in recombinant protein therapeutics.

Objective To investigate the predictive value of the combined tumor burden score (TBS) and platelet-albumin-bilirubin (PALBI) score model for postoperative tumor recurrence in liver transplant recipients with hepatocellular carcinoma (HCC). Methods The general information of 158 recipients diagnosed with HCC and underwent liver transplantation at the 900th Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army from 2008 to 2021 was collected. Lasso regression analysis combined with multivariate Cox regression analysis were used to identify independent risk factors for postoperative tumor recurrence after liver transplantation with HCC. A nomogram prediction model was constructed based on variables selected by Lasso regression analysis, and the predictive performance of the model was verified by calibration curve and clinical decision curve. The optimal cut-off values for postoperative tumor recurrence in liver transplant recipients with HCC were determined by receiver operating characteristic (ROC) curve, and Kaplan-Meier analysis was used to compare survival differences among different groups. Results Among the 158 liver transplant recipients with HCC, 82 experienced tumor recurrence, with a recurrence rate of 51.9% and a median tumor-free survival time of 10 (4, 25) months. Results of Lasso regression analysis and multivariate Cox regression analysis showed that alpha-fetoprotein (AFP) ≥400 ng/mL, TBS and PALBI score were all independent risk factors for postoperative tumor recurrence in liver transplant recipients with HCC (all P<0.05). The combined high TBS-high PALBI score showed the highest predictive value (hazard ratio 6.909, 95% confidence interval 3.067-15.563, P<0.001). A nomogram prediction model was constructed based on six variables selected by Lasso regression analysis. Calibration curve showed good consistency between the model's predicted results and the ideal curve. Decision curve analysis indicated that the nomogram prediction model provided the highest clinical benefit for predicting 1-year tumor-free survival after liver transplantation with HCC. Time-dependent ROC curves at 1, 3 and 5 years after surgery showed that TBS-PALBI model had good predictive performance, with no significant difference in area under the curve (AUC) compared with TBS-PALBI-AFP model. The optimal cut-off values for predicting postoperative tumor recurrence were determined by ROC curve, with a PALBI score cut-off of −2.334 and a TBS cut-off of 5.305. Recipients were divided into a low TBS-low PALBI score group (n=47) and a low/high TBS-low/high PALBI score group (at least one score was high) (n=111). Kaplan-Meier survival analysis showed that the low TBS-low PALBI score group had a higher tumor-free survival rate than the low/high TBS-low/high PALBI score group, with a significant difference (P<0.05). Conclusions TBS-PALBI model provides a novel, simple and effective tool for assessing the prognosis of liver transplant recipients with HCC. The nomogram model constructed based on this has significant advantages in predictive performance and may serve as a reference for guiding individualized treatment plans and improving clinical outcomes.

Preimplantation polygenic testing， as a frontier technology in assisted reproduction， provides a new way to prevent genetic diseases and reduce birth defects. However， it may also trigger a series of ethical issues. Combined with the research progress of preimplantation polygenic testing， this paper explored the ethical issues that may be caused by this technology， including the risks and uncertain benefits for future offspring and the challenges of informed consent， the dilemmas of autonomous decision-making and genetic responsibility of prospective parents， as well as social risks such as the distortion of the view of life and health inequities. Given the technical safety issues and ethical challenges posed by preimplantation polygenic testing， it was proposed to revise the regulations related to preimplantation genetic testing， as well as improve the ethical review of preimplantation polygenic testing， genetic counseling services， and public education on genetic science and genetic ethics. These ethical countermeasures aim to prevent and mitigate risks and ensure that technology truly benefits human society.

Based on his extensive clinical experience and the team's mechanistic research, Professor XU Nenggui has proposed the academic concept that "the governor vessel governs the brain and treats cerebral viscus diseases", and established a novel acupuncture approach for encephalopathy treatment centered on the integrated theory of "governor vessel-brain-mind", and developed a staged acupuncture protocol of "governor vessel regulating spirit" for ischemic stroke. This article introduces the academic features of this method in treating post-stroke dysphagia from four aspects: theoretical framework, treatment principles and point selection, mechanistic research, and clinical case studies. In clinical application, the method emphasizes syndrome differentiation based on meridians, harmonization between the conception and governor vessels; precise acupoint selection to treat both form and spirit; stage-specific differentiation with targeted needling. Furthermore, Professor XU integrates basic research with clinical practice, focusing on the neurobiological mechanisms underlying the efficacy of acupuncture in treating post-stroke dysphagia.
Acupuncture Therapy/methods* ; Humans ; Deglutition Disorders/psychology* ; Stroke/complications* ; Meridians ; Acupuncture Points

Two novel diketopiperazines (1 and 5), along with ten known compounds (2-4, 6-12) demonstrating significant skin inflammation inhibition, were isolated from a marine-derived fungus identified as Aspergillus sp. FAZW0001. The structural elucidation and configurational reassessments of compounds 1-5 were established through comprehensive spectral analyses, with their absolute configurations determined via single crystal X-ray diffraction using Cu Kα radiation, Marfey's method, and comparison between experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 1, 2, and 8 exhibited significant anti-inflammatory activities in Propionibacterium acnes (P. acnes)-induced human monocyte cell lines. Compound 8 demonstrated the ability to down-regulate interleukin-1β (IL-1β) expression by inhibiting Toll-like receptor 2 (TLR2) expression and modulating the activation of myeloid differentiation factor 88 (MyD88), mitogen-activated protein kinase (MAPK), and nuclear factor κB (NF-κB) signaling pathways, thus reducing the cellular inflammatory response induced by P. acnes. Additionally, compound 8 showed the capacity to suppress mitochondrial reactive oxygen species (ROS) production and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome activation, thereby reducing IL-1β maturation and secretion. A three-dimensional quantitative structure-activity relationships (3D-QSAR) model was applied to compounds 5-12 to analyze their anti-inflammatory structure-activity relationships.
Humans ; Aspergillus/chemistry* ; Diketopiperazines/isolation & purification* ; Anti-Inflammatory Agents/isolation & purification* ; Interleukin-1beta/genetics* ; Toll-Like Receptor 2/immunology* ; Propionibacterium acnes/drug effects* ; NF-kappa B/genetics* ; Molecular Structure ; Myeloid Differentiation Factor 88/immunology* ; Monocytes/immunology* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Cell Line

BACKGROUND:Current studies have confirmed that Ganoderma lucidum polysaccharides can promote nerve regeneration in neurodegeneration-related diseases.The occurrence of neurodegenerative diseases is closely related to mitochondrial dysfunction,but the role of Ganoderma lucidum polysaccharides on the regulation of apoptosis and mitochondrial function in neurodegenerative diseases is not yet clarified. OBJECTIVE:To explore the regulatory effects and mechanisms of Ganoderma lucidum polysaccharides on apoptosis and mitochondrial dysfunction in H2O2-induced SH-SY5Y cells. METHODS:SH-SY5Y cells were divided into three groups:control group,H2O2 group,and Ganoderma lucidum polysaccharides group.Cells in the control group were normally cultured.Cells in the H2O2 group were treated with 300 μmol/L H2O2 for 24 hours.In the Ganoderma lucidum polysaccharides group,the intervention with 300 μg/L Ganoderma lucidum polysaccharides was conducted first for 1-2 hours,followed by the addition of 300 μmol/L H2O2 for 24 hours.The mitochondrial membrane potential was detected by JC-1 kit.Apoptosis was detected by TUNEL staining kit.The activities of malondialdehyde and superoxide dismutase were detected by malondialdehyde test kit and superoxide dismutase test kit,respectively.The apoptosis and expression of mitochondrial dynamics-related proteins were detected by immunofluorescence staining and western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the mitochondrial membrane potential and superoxide dismutase activity were significantly reduced,as well as apoptotic rate and malondialdehyde levels were significantly increased in the H2O2 group(P<0.05).After treatment with Ganoderma lucidum polysaccharides,the membrane potential and superoxide dismutase activities were significantly increased,and apoptotic rate and malondialdehyde levels were significantly reduced compared with the H2O2 group(P<0.05).(2)The expression levels of pro-apoptotic proteins Bax and Caspase-3 were significantly increased,but the expression of anti-apoptotic protein Bcl-2 was significantly decreased in the H2O2 group compared with the control group(P<0.05).Compared with the H2O2 group,the levels of Bax and Caspase-3 were significantly decreased,but the expression of anti-apoptotic protein Bcl-2 was significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(3)Compared with the control group,the expression of mitochondrial splitting proteins Fis1 and p-Drp1 was significantly increased,but the expression of mitochondrial fusion proteins OPA1,Mfn1,and Mfn2 was decreased in the H2O2 group(P<0.05).Compared with the H2O2 group,Fis1 and p-Drp1 expression was significantly reduced,but the expression levels of OPA1,Mfn1,and Mfn2 were significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(4)The above results confirm that Ganoderma lucidum polysaccharides can attenuate H2O2-induced oxidative stress damage and apoptosis in SH-SY5Y cells by ameliorating mitochondrial dysfunction.

Donation after death is the most important ethical principle to carry out organ donation after citizens’ death. The newly-revised Regulations on Human Organ Donation and Transplantation does not define death, and avoids the key question of “whether to recognize brain death”. Certain legal risks or damages to the rights and interests of donors may exist in organ donation. Death is an inevitable part of human life. It is necessary to establish specific criteria, which is also the only approach, to define death in any era. Death criteria are established based on the view of death, and restricted by the development level of productive forces and other social factors. The determination of death criteria hugely varies between China and the West. To standardize organ donation and transplantation and promote high-quality development of organ donation, medical staff must adhere to the principle of pure motivation, take informed consents as the premise, respect the donors' and their close relatives' rights to choose their own death criteria, strictly follow the death judgment procedures and operating norms, and ensure the scientificity, accuracy and fairness of death determination.

Objective:To investigate the predictive value of preoperative γ-glutamyl transferase/lymphocyte count ratio (GLR) levels for postoperative tumor recurrence in liver transplant recipients with liver cancer.Methods:The clinical data of 158 recipients who were diagnosed with hepatocellular carcinoma (hereinafter referred to as liver cancer) and received liver transplantation at the No. 900 Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army from January 2008 to October 2022 were retrospectively analyzed. X-tile software, the Kaplan-Meier method, univariate and multivariate Cox regression, and other statistical methods were performed. The predictive value of preoperative GLR levels for postoperative tumor recurrence in liver transplant recipients with liver cancer and the risk factors for tumor recurrence in liver cancer patients post-liver transplantation were analyzed.Results:The X-tile software analysis confirmed that 96.8 was the optimal cutoff value for the preoperative GLR level to predict recurrence. The grouping threshold for survival analysis using the GLR cutoff value was 96.8. The tumor recurrence rates at 1, 3, and 5 years after surgery in the low-level GLR group (90 cases) and the high-level GLR group were 19.3% vs. 44.2%, 31.8% vs. 60.0%, and 34.1% vs. 62.9% (68 cases), respectively, and the differences were statistically significant between the two groups ( P<0.05). The Kaplan-Meier survival curve analysis results showed that the overall postoperative survival rate and recurrence-free survival rate were significantly lower in the high-level GLR group than the low-level GLR group ( P<0.05). The univariate Cox analysis result showed that there were statistically significant differences in preoperative aspartate aminotransferase, alpha fetoprotein, surgery time, maximum diameter of a solitary tumor, presence or absence of microvascular invasion, presence or absence of portal vein tumor thrombus, and preoperative GLR levels between the two groups ( P<0.05). Multivariate Cox analysis results showed that preoperative alpha-fetoprotein ≥400 ng/ml, GLR≥96.8, and the maximum diameter of a solitary tumor ≥5.0 cm were independent risk factors for postoperative tumor recurrence in liver transplant recipients with liver cancer ( P<0.05). Conclusion:GLR levels have a certain predictive value for postoperative tumor recurrence in liver transplant recipients with liver cancer. Furthermore, the postoperative tumor recurrence rate is relatively high when the preoperative GLR level in liver transplant recipients with liver cancer is ≥96.8.

Hepatolenticular degeneration is kind of an autosomal recessive genetic disease with diverse, complex and non-specific clinical manifestations, high misdiagnosis rate, rapid disease progression, poor drug treatment effect, and high mortality. It is one of the rare several genetic metabolic diseases in clinic that could be cured by liver transplantation method. Liver transplantation provides healthy P-type ATP enzyme through the donor liver, which can correct its genetic defects, improve copper metabolism disorders, relieve clinical symptoms, improve the quality of life, and improve the survival rate of patients. Liver transplantation is playing an increasingly important role as an important means to treat hepatolenticular degeneration. With the rapid development of partial living donor liver transplantation, auxiliary liver transplantation, domino-assisted liver transplantation and cross-assisted domino liver transplantation, a new way has been provided for patients with hepatolenticular degeneration, alleviating the problem of donor liver shortage and shortening the waiting time of recipients, which has certain clinical value and development prospects. In this paper, a review of the research progress in the treatment of hepatolenticular degeneration with liver transplantation was made with reference to the relevant literature at home and abroad.

Objective To investigate the effects of different sterilization methods on the quality of Astragali Radix decoction pieces by comparing the properties,odor,fingerprint,sterilization effect,extract and active component contents of Astragali Radix decoction pieces,and to further optimize the best sterilization process parameters of Astragali Radix decoction pieces.Methods The effects of moist heat sterilization,irradiation sterilization and dry heat sterilization on the quality of Astragali Radix decoction pieces were compared,and the best sterilization method was selected.Based on this,taking the content of astragaloside IV,the content of isoflavone glucoside,the total content of the two and the sterilization rate as the assessment indicators,taking the sterilization temperature,sterilization time and material thickness as the assessment factors,on the basis of the single-factor test,the response surface method was used to optimize the optimal dry heat sterilization process of Astragali Radix decoction pieces.Results The dry heat sterilization method was selected to sterilize Astragali Radix decoction pieces.The best process for dry heat sterilization was as follows:the sterilization temperature of 113℃,sterilization time of 5.1 h,and the material thickness of 19 mm.Conclusion Considering the sterilization effect,the influence on the content of active ingredients,operability and cost,selecting dry heat sterilization is the best sterilization method for Astragali Radix decoction pieces,and the optimal sterilization process is stable and feasible,with good repeatability,which can provide scientific basis for its industrial production application.