Objective: To establish a real-time quality control scheme based on the median (MD-IC) of internal control cycle threshold value in negative samples (NEG-IC-CT), so as to monitor anomalies such as progressive drift in nucleic acid testing system not covered by conventional internal quality control (IQC) in blood center nucleic acid laboratories, and to verify its feasibility. Methods: The internal control CT values of 54 426 negative samples were retrospectively collected. These samples were from four reagent batches of the two new and old equipment sets during the operation of the Wantai nucleic acid testing system in our blood center. The daily median of NEG-IC-CT values was used as the research indicator. Control limits were calculated using median absolute deviation (MAD) to construct the Median-MAD quality control chart. The monitoring performance of this scheme for the operation status of the testing system was simultaneously evaluated. Results: Statistical analysis showed significant differences in NEG-IC-CT value distribution between the new and old equipment sets, as well as between the two different reagent batches of the old equipment (P<0.000 1). The NEG-IC-CT value performance of the two different reagent batches of the new equipment was no significant difference in distribution (P>0.05). This scheme identified three typies of distinct anomalies. The out-of-control events observed with the old equipment in both the O1 and O2 reagent batches suggested potential performance decay due to equipment aging. The unreported change of reagent batch in time of Phase B with new equipment caused a stepwise drift on the quality control chart. In the later stage of Phase A with the new equipment, an alert was triggered, indicating potential quality risks associated with practices such as the mixed use of the remaining reagents and extremely long operator working hours. Conclusion: The realtime quality control scheme based on NEG-IC-CT value established in this study has been preliminarily validated for its monitoring effectiveness in nucleic acid testing in our blood center. This scheme performed well in detecting differences among testing systems and reagent batches, serving as an effective supplement to routine internal quality control. It can provide an intuitive and effective evaluation method for monitoring the performance of the nucleic acid testing process at blood center.

BACKGROUND: Epidermal growth factor receptor (EGFR) sensitive mutation is one of the effective targets of targeted therapy for non-small cell lung cancer (NSCLC). However, due to the difficulty of obtaining some primary tissues and the economic factors in some underdeveloped areas, some patients cannot undergo traditional genetic testing. The aim of this study is to establish a machine learning (ML) model using non-invasive peripheral blood markers to explore the biomarkers closely related to EGFR mutation status in NSCLC and evaluate their potential prognostic value. METHODS: 2642 lung cancer patients who visited Jiangsu Cancer Hospital from November 2016 to May 2023 were retrospectively enrolled and finally 175 NSCLC patients with complete follow-up data were included in the study. The ML model was constructed based on peripheral blood indicators and divided into training set and test set according to the ratio of 8:2. Unsupervised learning algorithms were used for clustering blood features and mutual information method for feature selection, and an ensemble learning algorithm based on Shapley value was designed to calculate the contribution of each feature to the model prediction result. The receiver operating characteristic (ROC) curve was used to evaluate the predictive ability of the model. RESULTS: Through the feature extraction and contribution analysis of the predictive results of the interpretable ML model based on the Shapley value, the top ten indicators with the highest contribution were: pathological type, phosphorus, eosinophils, monocyte count, activated partial thromboplastin time, potassium, total bilirubin, sodium, eosinophil percentage, and total cholesterol. The area under the curve (AUC) of the model was 0.80. In addition, patients with hyponatremia and squamous cell carcinoma group had a poor prognosis (P<0.05). CONCLUSIONS The interpretable model constructed in this study provides a new approach for the prediction of EGFR mutation status in NSCLC patients, which provides a scientific basis for the diagnosis and treatment of patients who cannot undergo genetic testing.
Humans ; Carcinoma, Non-Small-Cell Lung/diagnosis* ; Machine Learning ; Lung Neoplasms/diagnosis* ; Male ; Female ; Mutation ; Middle Aged ; ErbB Receptors/genetics* ; Prognosis ; Aged ; Retrospective Studies ; Adult ; Biomarkers, Tumor/genetics*

Pseudorabies (PR) is an infectious disease caused by the pseudorabies virus (PRV), affecting various domesticated and wild animals. Since pigs are the only natural hosts of PRV, PR poses a serious threat to the pig farming industry. Currently, PR is primarily prevented through vaccination with inactivated vaccines or genetically modified attenuated live vaccines. Developing safe and effective genetically engineered vaccines would facilitate the eradication and control of PR. In this study, the PRV vaccine strain Bartha-K61 was used as the reference strain. The gB protein was expressed via the baculovirus-insect cell expression system. Non-denaturing gel electrophoresis confirmed that the gB protein could form a trimeric structure. The purified gB protein was used to immunize mice, and the immune effect was evaluated by a challenge test. The results showed that the gB antigen induced a strong immune response in mice, with the serum-neutralizing antibody titer above 1:70. The lymphocyte stimulation index reached more than 1.29, and the level of (interferon gamma, IFN-γ) release was higher than 100 pg/mL. After immunization, mice were challenged with the virus at a dose of 10⁴ TCID₅₀/mL, 200 μL per mouse, and the clinical protection rate was 100%. Immunohistochemistry, histopathological section, and tissue viral load results showed that the pathological damage and viral load in the gB-immunized group were significantly lower than those in the PBS group. In summary, the gB protein obtained in this study induced strong humoral and cellular immune responses in mice, laying a foundation for developing a recombinant gB protein subunit vaccine.
Animals ; Mice ; Baculoviridae/metabolism* ; Viral Envelope Proteins/biosynthesis* ; Herpesvirus 1, Suid/genetics* ; Pseudorabies/immunology* ; Swine ; Pseudorabies Vaccines/genetics* ; Antibodies, Viral/blood* ; Insecta/cytology* ; Mice, Inbred BALB C ; Female ; Viral Vaccines/immunology*

Objective To construct a differential diagnosis model for systemic lupus erythematosus(SLE)from other autoimmune diseases based on the conventional test indicators in clinical laboratory,so as to improve the diagnostic efficacy of the existing test indicators.Methods The data of 178 SLE patients(SLE group)and 196 patients with other autoimmune diseases(control group)diagnosed in Sichuan Provincial People's Hospital from Apr.2022 to Mar.2023 were retrospectively analyzed.The differences in the levels of 19 clinical routine indicators between the 2 groups were analyzed.The least absolute shrinkage and selection operator(LASSO)regression was used to screen for test indicators with non-zero coefficients.These indicators were then used in logistic regression to construct a Nomogram model for SLE differential diagnosis.Model performance was assessed using receiver operating characteristic(ROC)curves and decision curve analysis.Results The levels of anti-cardiolipin antibody immunoglobulin(Ig)G,anti-cardiolipin antibody IgA,high-sensitivity C reactive protein(hs-CRP),D-dimer,and thrombin time(TT)in the SLE group were significantly higher than those in the control group(all P＜0.05),while the levels of IgM,complement 3(C3),complement 4(C4),prothrombin time(PT),and activated partial thromboplastin time(APTT)in the SLE group were significantly lower than those in the control group(all P＜0.05).Through LASSO regression,IgM,C3 and C4 were selected as the most likely indicators with non-zero coefficients.Multivariate logistic regression analysis showed that the differential diagnosis model was Logit P=4.18-1.34 × IgM-1.70 × C3-6.61 × C4.The area under the curve of this model was 0.80(95％confidence interval 0.76-0.85),with a sensitivity of 0.77 and a specificity of 0.74.Decision curve analysis demonstrated favourable clinical utility within a threshold probability range of 0.2-0.9.Conclusion The present model,constructed using the clinical routine indicators,such as IgM,C3 and C4,is helpful for the differential diagnosis of SLE from other autoimmune diseases and has good clinical application value.

Objective To systematically compare the perioperative outcomes of robot-assisted transvaginal natural orifice transluminal endoscopic surgery total hysterectomy(R-vNOTES-TH),transvaginal natural orifice transluminal endoscopic surgery total hysterectomy(vNOTES-TH),and robot-assisted laparoendoscopic single-site total hysterectomy(R-LESS-TH),and to evaluate the clinical advantages of R-vNOTES-TH.Methods Clinical data of 259 patients undergoing total hysterectomy for benign diseases at Zhongnan Hospital of Wuhan University from Jan.2020 to Dec.2024 were retrospectively analyzed.Baseline indicators and perioperative indicators were collected.Patients were assigned to 3 groups according to the surgical approach:R-vNOTES-TH group(n=22),vNOTES-TH group(n=39),or R-LESS-TH group(n=198).Perioperative indicators were compared between the R-vNOTES-TH group and the other 2 groups to evaluate the advantages of R-vNOTES-TH.Results Compared with the vNOTES-TH group,the R-vNOTES-TH group had significantly less intraoperative blood loss(50[50,100]mL vs 100[50,100]mL,P=0.027),lower intraoperative fluid infusion volume(1 000[500,1 000]mL vs 1 000[1 000,1 500]mL,P＜0.001),and shorter urinary catheter indwelling time(3[1,4]d vs 5[2,5]d,P=0.043),but longer vaginal drain indwelling time(2[2,3]d vs 2[0,2]d,P=0.004).Compared with the R-LESS-TH group,the R-vNOTES-TH group had longer urinary catheter indwelling time(3[1,4]d vs 1[1,1]d,P＜0.001).Conclusion Compared with vNOTES-TH,R-vNOTES-TH enhances intraoperative operational precision,reduces bleed loss,and accelerates urinary catheter removal,confirming that the robotic system effectively overcomes the technical limitations of conventional vNOTES.Although R-vNOTES-TH eliminates abdominal wall trauma-thereby prolonging urinary catheter indwelling time relative to R-LESS-TH-it offers patients a truly scar-free alternative.

Objective To investigate the effect of abnormal iron metabolism on neurological func-tion in elderly patients with hypertensive intracerebral hemorrhage(HICH)after minimally inva-sive surgical evacuation.Methods A prospective study was conducted on 300 elderly patients with HICH admitted to our hospital from January 2021 to December 2023.At 28 d after surgery,Glas-gow Outcome Scale(GOS)was used to assess the presence of neurological deficits or not,and then they were divided into a good neurological function group(GOS score≥4,175 cases)and a poor neurological function group(GOS score＜4,125 cases).Iron deposition in cerebrospinal fluid and serum iron metabolism were compared between the two groups,and the risk factors for neu-rological deterioration were analyzed.Results Compared with the good neurological function group,the poor neurological function group exhibited significantly decreases in Glasgow coma scale(GCS)scores at admission and 7 d after admission and iron ions(P＜0.01).Bleeding vo-lume,ferritin,transferrin,and quantitative susceptibility mapping(QSM)values of the thalamus and the hippocampus were obviously increased upon admission(P＜0.01).Multi variate logistic regression analysis showed that admission bleeding volume(OR=1.083,95％CI:1.012-1.159,P=0.021),ferritin(OR=1.065,95％CI:1.016-1.116,P=0.009),and thalamic QSM value(OR=4.075,95％CI:2.848-5.830,P=0.000)were risk factors for neurological dysfunction in the HICH patients after minimally invasive surgical treatment,while GCS score(OR=0.430,95％CI:0.259-0.715,P=0.001)and iron ions(OR=0.193,95％CI:0.064-0.581,P=0.003)at 7 d of admission were protective factors.Conclusion Iron deposition in cerebrospinal fluid and abnormal iron metabolism in serum are related to the deterioration of neurological function in eld-erly patients with HICH after minimally invasive hematoma evacuation,and are regarded as po-tential therapeutic targets.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:To determine the median effective dose (ED 50) of oliceridine inhibiting responses to insertion of the gastroscope when combined with propofol in obese patients. Methods:In this prospective study, American Society of Anesthesiologists Physical Status classification I or Ⅱ patients of either sex, with a body mass index of ≥28 kg/m 2, undergoing elective painless gastroscopy, were selected. Propofol 1.5 mg/kg was slowly injected intravenously 2 min after intravenous injection of oliceridine. Gastroscopy was performed after the patient′s eyelash reflex disappeared and the jaw was relaxed. The optimal dose was determined by modified Dixon′s up and down method. The initial dose of oliceridine was 0.015 mg/kg, and the ratio between the two successive concentrations was 1.0∶1.2. If the patient had a positive response that affected the examination operation such as swallowing, bucking and body movement during insertion of the gastroscope, the next patient received a higher dose, or conversely if negative, a lower dose was given in the next patient. This process was repeated until the 7th turning point occurred. The ED 50 and 95% confidence interval of olicerdine inhibiting responses to insertion of the gastroscope were calculated by probit method. Results:A total of 21 patients were included in this study, the ED 50 of olicerdine inhibiting responses to implantation of the gastroscope was 0.016 mg/kg, and the 95% confidence interval was 0.010-0.023 mg/kg when combined with propofol 1.5 mg/kg in obese patients. One patient developed hypoxemia. One patient had mild dizziness after operation. Conclusions:The ED 50 of olicerdine inhibiting responses to implantation of the gastroscope is 0.016 mg/kg when combined with propofol 1.5 mg/kg in obese patients.

Objective:To evaluate the effect of sevoflurane on mitochondrial-associated endoplasmic reticulum membranes (MAMs) in mouse microglia and the relationship with silent information regulator-related enzyme 3 (SIRT3)/adenosine triphosphatase family protein 3A (ATAD3A) signaling pathway.Methods:After normal culture of mouse microglia (BV-2 cell line), the cells were divided into 4 groups ( n=33 each) by a random number table method: empty adenovirus-control group (group C), empty adenovirus-sevoflurane exposure group (group Sev), SIRT3 overexpression adenovirus-control group (group SIRT3 + C) and SIRT3 overexpression adenovirus-sevoflurane exposure group (group SIRT3 + Sev). The cells were infected with SIRT3 overexpression adenovirus 100 pmol/well in SIRT3+ C group and SIRT3+ Sev group and with empty adenovirus 100 pmol/well in C group and Sev group. After 48 h of infection, the cells were routinely cultured for 48 h in C group and SIRT3+ C group, the cells were incubated with 3% sevoflurane for 2 h, once a day for 3 consecutive days, followed by routine culture for 48 h in Sev group and SIRT3+ Sev group. The contents of mitochondrial Ca 2+ and reactive oxygen species (mtROS) were measured by flow cytometry. The mitochondrial membrane potential (MMP) was measured by JC-1 probe. The mitochondrial ATP content was measured by luciferase luminescence method. The expression of SIRT3 was detected by Western blot. The expression of acetylated ATAD3A was detected by immunoprecipitation. The co-localization of endoplasmic reticulum and mitochondria was determined by confocal fluorescence microscopy, and the Manders co-localization coefficient was calculated to evaluate the development of MAMs. Results:Compared with group C, the contents of mitochondrial Ca 2+ and mtROS were significantly increased, the contents of MMP and mitochondrial ATP were decreased, the expression of SIRT3 was down-regulated, the expression of acetylated ATAD3A was up-regulated, and the development of MAMs was increased in group Sev ( P<0.05). Compared with Sev group, the contents of mitochondrial Ca 2+ and mtROS were significantly decreased, the contents of MMP and mitochondrial ATP were increased, the expression of SIRT3 was up-regulated, the expression of acetylated ATAD3A was down-regulated, and the development of MAMs was decreased in SIRT3 + Sev group ( P<0.05). Compared with SIRT3+ C group, the contents of mitochondrial Ca 2+ and mtROS were significantly increased, the contents of MMP and mitochondrial ATP were decreased, the expression of SIRT3 was down-regulated, the expression of acetylated ATAD3A was up-regulated, and the development of MAMs was increased in SIRT3+ Sev group ( P<0.05). Conclusions:The mechanism by which sevoflurane induces mitochondrial dysfunction in mouse microglia may be related to the inhibition of SIRT3/ATAD3A signaling pathway, leading to excessive development of MAMs.

Objective:To determine the median effective dose (ED 50) of oliceridine inhibiting responses to insertion of the gastroscope when combined with propofol in obese patients. Methods:In this prospective study, American Society of Anesthesiologists Physical Status classification I or Ⅱ patients of either sex, with a body mass index of ≥28 kg/m 2, undergoing elective painless gastroscopy, were selected. Propofol 1.5 mg/kg was slowly injected intravenously 2 min after intravenous injection of oliceridine. Gastroscopy was performed after the patient′s eyelash reflex disappeared and the jaw was relaxed. The optimal dose was determined by modified Dixon′s up and down method. The initial dose of oliceridine was 0.015 mg/kg, and the ratio between the two successive concentrations was 1.0∶1.2. If the patient had a positive response that affected the examination operation such as swallowing, bucking and body movement during insertion of the gastroscope, the next patient received a higher dose, or conversely if negative, a lower dose was given in the next patient. This process was repeated until the 7th turning point occurred. The ED 50 and 95% confidence interval of olicerdine inhibiting responses to insertion of the gastroscope were calculated by probit method. Results:A total of 21 patients were included in this study, the ED 50 of olicerdine inhibiting responses to implantation of the gastroscope was 0.016 mg/kg, and the 95% confidence interval was 0.010-0.023 mg/kg when combined with propofol 1.5 mg/kg in obese patients. One patient developed hypoxemia. One patient had mild dizziness after operation. Conclusions:The ED 50 of olicerdine inhibiting responses to implantation of the gastroscope is 0.016 mg/kg when combined with propofol 1.5 mg/kg in obese patients.