ObjectiveTo investigate the functional role and underlying molecular mechanisms of fumarylacetoacetate hydrolase （FAH） in the progression of glioblastoma （GBM）. MethodsDifferential expression analysis was performed on the TCGA-GBM， GSE4290， and GSE116520 datasets. Weighted gene co-expression network analysis （WGCNA） was used to identify key modules， and Cox regression and risk modeling were used to screen prognostic genes. Immune infiltration analysis of prognostic genes was carried out by using single-cell RNA sequencing panels. The clinical expression signature of FAH in GBM was analyzed in the TCGA and HPA databases. The functional role of FAH was validated by in vitro and in vivo experiments， and pathway analysis was performed to explore the underlying mechanisms. ResultsA total of 152 overlapping genes were identified across the three GBM datasets （P<0.05）. WGCNA revealed that the turquoise module was most strongly associated with tumor purity， stromal score， immune score， and ESTIMATE score （P<0.001）. Compared with normal tissues， three prognostic genes （CTSD， FAH， and THBD） were upregulated in GBM and correlated with immune infiltration （P<0.05）. FAH mRNA and protein levels were elevated in GBM tissues relative to normal tissues， and its expression was significantly associated with age stratification and TP53 mutation （P<0.05）. CCK-8 assay results showed that， compared with the shNC group， the proliferative activity of GBM cells in the shFAH group was reduced （P<0.001）. Transwell migration and invasion assays demonstrated that， relative to the shNC group， the numbers of migrated and invaded cells in the shFAH group decreased （P<0.05）. Western blot analysis revealed that the protein expression levels of PI3K， p-AKT， and p-mTOR in the shFAH group decreased compared with those in the shNC group （P<0.05）. In vivo subcutaneous xenograft experiments further confirmed that tumor volume and weight significantly decreased in the shFAH group compared with the shNC group （P<0.001）. ConclusionFAH promotes GBM progression by activating the PI3K/AKT/mTOR signaling pathway and may serve as a potential therapeutic target for GBM.

Objective:To analyze the clinical manifestations, genetic and hematological test results of patients with sitosteronism (STSL) complicated with cardiovascular and cerebrovascular events.Methods:Clinical data were collected from 11 STSL patients at the outpatient department of Ruijin Hospital affiliated to Shanghai Jiaotong University School of Medicine between November 2020 to June 2023. The whole exome sequencing technology was used to detect gene mutations associated with lipid metabolism, the serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were tested by the enzyme endpoint method; serum phytosterol levels by high-performance liquid chromatography; serum concentration of C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) by enzyme-linked immunosorbent assay, concentration of fibrinogen, the activity of protein C and coagulation factor Ⅷ (FⅧ) by the coagulation method; the antigen and activity of von Willebrand factor (vWF) by immunoturbidimetric assay; and the activity of antithrombin Ⅲ (ATⅢ) by chromogenic substrate assay.Results:There were 3 cases of coronary heart disease, 6 cases of cerebral infarction, 2 cases of coronary heart disease combined with cerebral infarction, 4 cases of eyelid melasma, and 2 cases of arthritis. Gene mutation was as follows: ABCG5 gene mutation including exon9: c G1166A: p R389H, exon9: c T1195C: p F399L, exon12: c.1762+1G>A, and ABCG8 gene mutation including exon 11 c.1720G>A: p.Gly574Arg, exon4:c.445_453del:p.A149_V151del, exon13 c.1949T>G: p.Leu650Arg. The percent of stomatocytes in the peripheral blood swears was (11.3±8.6)%. The concentrations of TC, LDL-C and sitosterol was (6.8±2.4), (4.4±2.0) mmol/L and 40.0 (22.0, 203.7) μmol/L. The level of CRP, interleukin IL-6, and TNF-α was 15.5 (7.2, 29.6)mg/L, (4.2±2.0) pg/ml and (6.7±1.5) pg/ml, respectively.The activity of PC, FⅧ and ATⅢ was (114±51)%, (110±41)% and (83±33)%. The values of FIB was (3.2±1.4)g/L.vWF antigen and vWF activity was (305±168)% and (275±112)%.Conclusions:STSL patients combined with cardiovascular and cerebrovascular events not only had complex dysfunctional lipid metabolism related gene defects, but also had significantly increased hematological indicators such as inflammatory mediator CRP, coagulation parameter vWF.

OBJECTIVE To prepare Lanqin extract/cyclodextrin complexes for probing its effects of different kinds of cyclodextrins on the taste-masking. METHODS Bitter compounds in the extracts were performed on ion exchange resin adsorption combined with HPLC. The formulations of complexes were screened by human taste panel method. The complexes were prepared by spray-drying and characterized through scanning electron microscope, differential scanning calorimetry, and hygroscopicty test. Moreover, the in vitro bitter taste perception of complexes was evaluated by electronic tongue and further valuation the credibility of the results was conducted on human gustatory sensation tests. RESULTS The sulfobutylether-β-cyclodextrin-based combinational formulation with multiple cyclodextrins could significantly inhibit the bitter taste of the extract which mainly caused by its alkaline constituents at a lower dosage. The results of electron scanning microscopy, differential scanning calorimetry, and hygroscopicity indicated that the Lanqin extract and cyclodextrin in the complex may form inclusion complexes rather than physical mixtures. The results of electronic tongue and human gustatory sensation tests showed that, compared with the extract suggested the taste characteristics of the optimal complexes was similar to corresponding excipient while the bitterness significantly reduced. CONCLUSION The Lanqin extract/cyclodextrin complexes prepared in this study are suitable for industrial production for its good flavour, less total amount of cyclodextrins, and simple process. The present study has important significance for the development of related taste masking products of Lanqin.

Objective:To evaluate the performance of an artificial intelligent (AI)-based automated digital cell morphology analyzer (hereinafter referred as AI morphology analyzer) in detecting peripheral white blood cells (WBCs).Methods:A multi-center study. 1. A total of 3010 venous blood samples were collected from 11 tertiary hospitals nationwide, and 14 types of WBCs were analyzed with the AI morphology analyzers. The pre-classification results were compared with the post-classification results reviewed by senior morphological experts in evaluate the accuracy, sensitivity, specificity, and agreement of the AI morphology analyzers on the WBC pre-classification. 2. 400 blood samples (no less than 50% of the samples with abnormal WBCs after pre-classification and manual review) were selected from 3 010 samples, and the morphologists conducted manual microscopic examinations to differentiate different types of WBCs. The correlation between the post-classification and the manual microscopic examination results was analyzed. 3. Blood samples of patients diagnosed with lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative neoplasms were selected from the 3 010 blood samples. The performance of the AI morphology analyzers in these five hematological malignancies was evaluated by comparing the pre-classification and post-classification results. Cohen′s kappa test was used to analyze the consistency of WBC pre-classification and expert audit results, and Passing-Bablock regression analysis was used for comparison test, and accuracy, sensitivity, specificity, and agreement were calculated according to the formula.Results:1. AI morphology analyzers can pre-classify 14 types of WBCs and nucleated red blood cells. Compared with the post-classification results reviewed by senior morphological experts, the pre-classification accuracy of total WBCs reached 97.97%, of which the pre-classification accuracies of normal WBCs and abnormal WBCs were more than 96% and 87%, respectively. 2. The post-classification results reviewed by senior morphological experts correlated well with the manual differential results for all types of WBCs and nucleated red blood cells (neutrophils, lymphocytes, monocytes, eosinophils, basophils, immature granulocytes, blast cells, nucleated erythrocytes and malignant cells r>0.90 respectively, reactive lymphocytes r=0.85). With reference, the positive smear of abnormal cell types defined by The International Consensus Group for Hematology, the AI morphology analyzer has the similar screening ability for abnormal WBC samples as the manual microscopic examination. 3. For the blood samples with malignant hematologic diseases, the AI morphology analyzers showed accuracies higher than 84% on blast cells pre-classification, and the sensitivities were higher than 94%. In acute myeloid leukemia, the sensitivity of abnormal promyelocytes pre-classification exceeded 95%. Conclusion:The AI morphology analyzer showed high pre-classification accuracies and sensitivities on all types of leukocytes in peripheral blood when comparing with the post-classification results reviewed by experts. The post-classification results also showed a good correlation with the manual differential results. The AI morphology analyzer provides an efficient adjunctive white blood cell detection method for screening malignant hematological diseases.

Objective:To investigate the effect of long-term oral administration of β-blocker on septic myocardial injury and prognosis.Methods:A retrospective study was conducted. Patients who were admitted to the emergency intensive care unit (EICU) and intensive care unit (ICU) of Tongde Hospital of Zhejiang Province from January 2015 to June 2020 were enrolled. A total of 289 patients who met the criteria of myocardial injury induced by sepsis were included in the analysis. Among them, 187 patients who had never taken β-blocker within 3 months before diagnosis were divided in the non-β-blocker group, and 102 patients who took β-blocker daily for more than 3 months before diagnosis were in the β-blocker group. The physiological and biochemical characteristics were compared between the two groups, including heart rate, mean arterial pressure (MAP) at the time of diagnosis, cardiac troponin I (cTnI), brain natriuretic peptide (BNP), MB isoenzyme of creatine kinase (CK-MB), blood lactic acid (Lac), central venous oxygen saturation (ScvO 2), sequential organ failure assessment (SOFA) score, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score within 24 hours of diagnosis, left ventricular ejection fraction (LVEF), early and late mitral orifice diastolic peak flow velocity ratio (E/A), utilization rate of vasoactive drugs during hospitalization and 28-day mortality. Results:The heart rate in the β-blocker group at the time of diagnosis was significantly lower than that in the non-β-blocker group (bpm: 107±8 vs. 110±7, P < 0.01), and the levels of cTnI and BNP within 24 hours of diagnosis were significantly lower than those in the non-β-blocker group [cTnI (μg/L): 0.191 (0.220) vs. 0.291 (0.300), BNP (ng/L): 627 (133) vs. 690 (201), both P < 0.05]. However, there were no significant differences in MAP, CK-MB, Lac, ScvO 2, SOFA score, APACHE Ⅱ score, LVEF, E/A, vasoactive drug utilization rate, and 28-day mortality between the β-blocker and non-β-blocker groups [MAP (mmHg, 1 mmHg = 0.133 kPa): 70.6±3.9 vs. 69.9±3.8, CK-MB (μg/L): 4.24 (3.33) vs. 4.32 (3.13), Lac (mmol/L): 3.50 (1.80) vs. 3.50 (1.90), ScvO 2: 0.729±0.032 vs. 0.735±0.041, SOFA score: 7.74±2.34 vs. 7.25±2.23, APACHE Ⅱ score: 17.19±5.13 vs. 18.27±6.12, LVEF: 0.567±0.058 vs. 0.557±0.051, E/A: 0.71 (0.20) vs. 0.69 (0.20), vasoactive drug utilization rate: 60.8% (62/102) vs. 56.7% (106/187), 28-day mortality: 23.5% (24/102) vs. 25.7% (48/187), all P > 0.05]. Conclusion:Long-term oral administration of β-blocker reduce myocardial injury in septic patients, and has no effect on disease severity and prognosis.

An 83-year-old male patient received moxifloxacin hydrochloride (moxifloxacin) 400 mg once daily orally for acute attack of chronic obstructive pulmonary disease. He developed scattered red rashes, accompanied by itching, on his both lower limbs 5 hours after the first dose. Next day, the rashes involved skin on the trunk, and purpura appeared on the multiple skin below the knees. Laboratory tests showed platelet count (PLT) 1×10 9/L, and thrombocytopenia related to moxifloxacin was considered. Moxi- floxacin was stopped and the treatments including hemostasis, anti-allergy, regulation of immune function, and platelet transfusion were given. On day 2 of drug withdrawal, his PLT was 3×10 9/L, and on day 4 the PLT was 35×10 9/L. He was transferred to a superior hospital and received the therapy including anti-immune response, platelet-raising, and hemostasis for 5 days. Then his PLT increased to 244×10 9/L.

An 83-year-old male patient received moxifloxacin hydrochloride (moxifloxacin) 400 mg once daily orally for acute attack of chronic obstructive pulmonary disease. He developed scattered red rashes, accompanied by itching, on his both lower limbs 5 hours after the first dose. Next day, the rashes involved skin on the trunk, and purpura appeared on the multiple skin below the knees. Laboratory tests showed platelet count (PLT) 1×10 9/L, and thrombocytopenia related to moxifloxacin was considered. Moxi- floxacin was stopped and the treatments including hemostasis, anti-allergy, regulation of immune function, and platelet transfusion were given. On day 2 of drug withdrawal, his PLT was 3×10 9/L, and on day 4 the PLT was 35×10 9/L. He was transferred to a superior hospital and received the therapy including anti-immune response, platelet-raising, and hemostasis for 5 days. Then his PLT increased to 244×10 9/L.

Objective: To establish a set of rules for autoverification of blood analysis, in order to provide a way to validate autoverification rules for different analytical systems, which can ensure the accuracy of test results as well as shorten turnaround time (TAT) of test reports. Methods: A total of 34 629 EDTA-K2 anticoagulated blood samples were collected from multicenter cooperative units including the First Hospital of Jinlin University during January 2017 to November 2017. These samples included: 3 478 cases in Autoverification Establishment Group, including 288 cases for Delta check rules; 5 362 cases in Autoverification Validation Group, including 2 494 cases for Delta check; 25 789 cases in Clinical Application Trial Group. All these samples were analyzed for blood routine tests using Sysmex XN series automatic blood analyzers.Blood smears, staining and microscopic examination were done for each sample; then the clinical information, instrument parameters, test results and microscopic results were summarized; screening and determination of autoverification conditions including parameters and cutoff values were done using statistical analysis. The autoverification rules were input into Sysmex Laboman software and undergone stage Ⅰ validation using simulated data, and stage Ⅱ validation for post-analytical samples successively. True negative, false negative, true positive, false positive, autoverification pass rate and passing accuracy were calculated. Autoverification rules were applied to autoverification blood routine results and missed detection rates were validated, and also data of autoverification pass rate and TAT were obtained. Results: (1)The selected autoverification conditions and cutoff values included 43 rules involving WBC, RBC, PLT, Delta check and abnormal characteristics. (2)Validation of 3 190 cases in Autoverification Establishment Group showed the false negative rate was 1.94%(62/3 190)(P<0.001), autoverification pass rate was 76.74%, passing accuracy was 97.47%; Validation of 2 868 cases in Autoverification Validation Group, the false negative rate was 3.38%(97/2 868)(P=0.002), autoverification pass rate was 42.26%, passing accuracy was 92.00%; Validation of Delta check on 288 cases in Autoverification Establishment Group and 2 494 cases in Autoverification Validation Group showed the false negative rates were respectively 1.39% and 2.61%(P<0.001). (3)Three hospitals adopted these rules of autoverification for 25 789 blood routine samples, and found that the average TAT of blood routine test reports were shortened by 24min, 32min and 7min respectively, the rate of samples reported within 30min were elevated by 33%, 53% and 7%. The autoverification pass rates were 72%-74%. Conclusions The application of this set of 43 autoverification rules in blood sample analysis can ensure test quality while shortenTAT and improve work efficiency. It is worth pointing out that for the same analytical systems in this research, validation is necessary before application of this set of rules, and periodic validation is required during application to make necessary adjustment; for different analytical systems, as this research provide a way to establish autoverification rules for blood routine tests.Clinical labs may establish their own suitable autoverification rules on the basis of technological parameters. (Chin J Lab Med, 2018, 41: 601-607)

objective To determine a proper fiducial photography distance setting for ideal amptitied endoscopic imaging of parathyroid gland by high definition endoscopy system.Methods 30 patients were operated with MIVAT mode (modified Miccoli's approach) for treatment of thyroid carcinoma from Apr.2013 to Mar.2014.High definition imaging was established by Image 1 Endoscopy System(Karl Storz Co.) to observe parathyroid gland and related fine anatomical structures during surgery.5 fiducial photography distances (1.0/1.5/2.0/2.5/3.0 cm) were separately tested during surgery.Maximally amplified parathyroid gland images of each setting were obtained by the approaching-amplifying photographic method,and then the size of the real parathyroid glands as well as their screen images were measured and recorded to calculate the magnification.A proper fiducial photography distance setting was determined postoperatively by comparison of the magnification times,as well as clarity,stability of the imaging and surgical maneuverability.Results ①90 parathyroid glands were successfully observed and measured.②At the longest fiducial photography distance (3.0 cm),the parathyroid gland could be stably magnified by 14.26±3.06(long trail)/12.62±2.88 (wide trail)times,but their contour and color not clear.③At the intermediate distance (2.5 cm),the parathyroid gland could be magnified by 16.74±3.15 (long trail)/14.81± 3.47(wide trail)times with the graphics stable,and the color and contour more clear,but the vascular pedicle and the tiny vessels under the capsule still blurred.④At the shortest distance (1.0 cm),the parathyroid gland could be magnified by 27.72±6.45 (long trail)/26.33±7.22(wide trail)times,not only the color and contour,but also the vascular pedicle and the tiny vessels under the capsule of the gland became further clearer,unfortunately the graphics was shimmy and unstable.Conclusions ①2.5 cm can be a proper fiducial photography distance for searching,identifying and preserving parathyroid gland in MIVAT,while 1.0 cm can be a special fiducial photography distance for further confirming parathyroid gland when necessary.② Current high definition endoscopy system can be applied to identify the parathyroid gland if fiducial photography distance was properly set and approachingamplifying photographic method was used.Along with the magnification of the imaging,the features of the parathyroid gland may become clearer,including its yellow-brown color and oval contour,as well as the detail structures such as the tiny vessels under the capsule and the vascular pedicle.

Hyperparathyroidism is an important complication of thyroid surgery.Identification is the premise of intraoperative pretection.At present,identification of the parathyroid gland relies on personal experience of surgeons.Amplifying display of endoscope or surgical magnifying glass,the use of dyeing agent such as methylene blue,nanocarbon,5-ALA or BB5-G1,the use of radionuclide imaging and contact endoscope,and biopsy like intraoperative frozen pathological examination and FNA are all important trials.This article is going to make a review of the methods.