Objective:To construct the liver organoid infected with hepatitis D virus (HDV), and to investigate the role of the sodium taurocholate cotransporting polypeptide (NTCP) receptor inhibitor bulevirtide in inhibiting viral replication.Methods:Hepatocyte-like cells (HLC) differentiated from induced pluripotent stem cells (iPSC) were seeded onto inverted colloidal crystal polyethylene glycol scaffolds (ICC) to construct liver organoids. After transfecting human hepatocelluar carcinoma cells (HuH7 cells) with plasmids, HDV particles were harvested from the supernatant, while HBV particles were extracted from the HepG2.2.15 cell supernatant. The liver organoids were infected with both HBV and HDV particles, and the negative control group without HDV infection was set up. The microstructure of the liver organoid units and the expression of hepatitis D antigen (HDAg) and hepatitis B surface antigen (HBsAg) were observed under laser scanning confocal microscope by immunofluorescence method. The protein levels of NTCP and HDAg in the liver organoids were detected by Western blotting. Bulevirtide was added before HDV infection (bulevirtide pre group) and 24 hours after infection (bulevirtide post group), and interferon-alpha (IFN-α) was also added after 24 hours infection (IFN-α group), and a control group without drug treatment was set up. HDV replication was compared among the four groups after drug intervention. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to measure the relative mRNA expression levels of Nanog homeobox (NANOG), sex determining region Y-box (SOX)2, SOX17, forkhead box protein A2 (FOXA2), hepatocyte nuclear factor 4 alpha (HNF-4α), albumin (ALB), alpha-fetoprotein (AFP), NTCP during the differentiation of iPSC, and the mRNA expression of HDV after the drug intervention of the four groups. Statistical analysis was performed using two independent sample t tests. Results:Within 21 days of the differentiation of iPSC into HLC, the mRNA expression level of NANOG gradually decreased, while the expression levels of SOX17, FOXA2 initially increased then decreased, and the expression levels of the HNF-4α, ALB, AFP and NTCP progressively increased. The protein level of NTCP in iPSC (0.118±0.003) was lower than that in HLC (1.315±0.073), and the difference was statistically significant ( t=11.92, P<0.001).The protein level of HDAg in the liver organoids after HDV infection was higher than that in the negative control group without HDV infection (1.284±0.128 vs 0.157±0.040), and the difference was statistically significant ( t=23.27, P<0.001).Laser scanning confocal microscopy showed three-dimensional spheroid structures and high expressions of HDAg and HBsAg at the 14th day of infection.Compared with the control group (1.000±0.077), the HDV mRNA expressions in both IFN-α group (0.453±0.028) and bulevirtide pre group (0.136±0.012) decreased after three days of drug intervention. The differences were statistically significant ( t=19.95 and 33.15, respectively, both P<0.001). However, there was no significant difference in HDV mRNA expressions between the bulevirtide post group (0.968±0.069) and the control group ( t=0.94, P>0.05). Conclusions:The liver organoids constructed from iPSC-derived HLC and ICC can simulate human liver functions and successfully be infected by HDV particles. Early blockade with bulevirtide can effectively reduce the level of viral replication in the HDV-infected liver organoids.

Objective To investigate the clinical features of of patients with Staphylococcus aureus blood-stream infection and risk factors for septic shock.Methods A total of 51 patients diagnosed with Staphylococ-cus aureus bloodstream infection in the hospital from January 2018 to March 2023 were enrolled in the study.According to whether the patients developed septic shock,they were divided into septic shock group and non-septic shock group.The clinical data of the patients were collected,and the clinical laboratory indicators were detected on the day of blood culture samples were collected.Bacteria isolated from blood culture specimens of patients were identified and tested for drug sensitivity.The clinical data and clinical laboratory test indicators of the two groups were compared.Multivariate Logistic regression was used to analyze the independent risk factors of septic shock in patients with Staphylococcus aureus bloodstream infection.The receiver operating characteristic(ROC)curve was used to analyze the predictive value of clinical laboratory test indicators for septic shock in patients with Staphylococcus aureus bloodstream infection.Results Septic shock occurred in 12 of 51 patients,with an incidence of 23.5％.The proportion of patients with diabetes,gouty arthritis,the proportion of patients with long-term glucocorticoid use,the proportion of patients with respiratory tract in-fection,the proportion of patients who died,and the hospitalization cost in the septic shock group were higher than those in the non-septic shock group,and the differences were statistically significant(P＜0.05).Long-term glucocorticoid use was an independent risk factor for septic shock in Staphylococcus aureus bloodstream infection(P＜0.05).The combination of C-reactive protein(CRP),albumin(Alb),neutrophil/lymphocyte ratio(NLR)and procalcitonin(PCT)had high value in predicting septic shock in patients with Staphylococ-cus aureus bloodstream infection,and the area under the ROC curve was 0.983.Conclusion Long-term use of glucocorticoids can lead to an increased risk of septic shock in patients with Staphylococcus aureus blood-stream infection.The combined detection of CRP,Alb,NLR and PCT has a higher predictive value than single detection for septic shock in patients with Staphylococcus aureus bloodstream infection.

Objective　To shorten the detection time of Staphylococcus aureus (SA), this study established a rapid detection method for the thermostable nuclease nuc gene and mecA resistance gene of SA based on recombinase aided amplification (RAA) combined with lateral flow strips (LFS). Methods Efficient RAA primers and probes were designed and screened based on the conserved sequences of the nuc gene in the SA genome and the mecA gene on the staphylococcal SCCmec removable genetic element, and then, the reaction temperature and for the simultaneous detection of nuc and mecA genes by time of RAA-LFS were verified. Sensitivity, specificity, and comparison with quantitative real-time PCR (qPCR) were also assessed. A prospective evaluation was conducted using 52 vials of non-repeat positive blood cultures from two tertiary hospitals. Results The RAA-LFS was able to amplify both nuc and mecA genes under the reaction conditions of 20-45 °C for 15-30 minutes, with a detection limit of 10² CFU/mL. A total of 50 clinically isolated non-SA strains were validated with 100% specificity for both nuc and mecA genes. Of the 30 methicillin-resistant Staphylococcus aureus (MRSA) and 30 methicillin-susceptible Staphylococcus aureus (MSSA) strains preserved in our laboratory, all were positive for the nuc gene, and 30 strains were positive for the mecA gene. The positive and negative concordance rates with qPCR were both 100%, with a consistency test Kappa value of 1. In a prospective analysis of the 52 vials of positive blood cultures, the identification and antibiotic susceptibility test results of 16 strains of MSSA and 6 strains of MRSA showed a 100% concordance rate with the results obtained using the Mérieux VITEK-2 compact microbiological detection system. Conclusions We combined nucleic acid release agents, RAA, and lateral flow strips to develop a simple, rapid, and highly sensitive assay applicable for SA and mecA resistance gene detection in colonies or positive blood culture bottles.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.

Objective:To explore the clinical efficacy of morcellator assisted by transurethral bipolar plasmakinetic enucleation and resction of the prostate in the treatment of benign prostatic hyperplasia.Methods:A total of 90 patients with benign prostatic hyperplasia treated in the Department of Urology, Shenyang Fifth People′s Hospital from Apirl 2020 to Apirl 2021 were retrospectively analyzed. They were divided into the experimental group ( n=50) and the control group ( n=40) according to different sugical methods. Among them, patients underwent transurethral plasma anatomical prostatectomy with the aid of morcellator in the experimental group, patients of the control group underwent plasma prostatectomy. The operation time, intraoperative blood loss, the decrease in hemoglobin, postoperative catheter retention, hospitalization time, international prostate symptom score (IPSS) of 3 months after surgery, quality of life score (QOL), maximum urine flow rate(Qmax), residual urine volume (PVR), surgical complications and other related indicators in the two groups were compared. Measurement data were expressed as mean±standard deviation ( Mean± SD), comparison between groups was by t-test; comparison of count data between groups was by Chi-square test. Results:Both groups were successfully treated by transurethral surgery. There were significant difference in the amount of blood loss [(62.11±5.32) mL vs (95.12±10.32) mL], the total operation time[(40.25±12.75) min vs (72.1±13.41) min], postoperative catheter retention[(3.02±0.41) d vs (4.73±1.32) d], hospitalization time[(4.03±0.41) d vs (6.52±0.85) d], the decrease in hemoglobin[(2.65±0.52) g/L vs (4.21±0.85) g/L]( P<0.05); there was no significant difference in the IPSS score(7.36±3.26 vs 8.12±3.56), QOL(2.12±0.32 vs 2.32±0.21), Qmax[(15.47±4.53) mL/s vs (16.23±3.21) mL/s], PVR [(15.25±5.14) mL vs (16.21±5.26) mL], the incidence of complications(6.00% vs 5.00%)( P>0.05). Conclusion:It is safe and feasible to treat benign prostatic hyperplasia by transurethral plasma anatomical prostatectomy assisted by tissue planer, which can significantly improve its clinical efficacy.

Objective:To examine the short-term outcomes of modified load-sharing rip-stop repair (mLSRS) and suture-bridge repair (SB) for large rotator cuff tear.Methods:The clinical data of patients who underwent shoulder arthroscopy for large rotator cuff repair at Department of Orthopedic, China Academy of Chinese Medical Sciences Wang Jing Hospital from January 2017 to December 2018 were collected. Totally 111 patients (39 in mLSRS group and 72 in SB group) were included in the study. There were 45 males and 66 females, aged (58.0±6.9)years (range: 40 to 72 years).A total of 39 pairs of patients were successfully matched by propensity score matching. The pain of shoulder joint was assessed by visual analogy score (VAS) , the shoulder function was assessed by Constant-Murley score, the abduction strength of bilateral shoulder joint was assessed by 90? force assessment method of upper limb on the plane of scapula, and the healing of rotator cuff on the operative side was assessed by sugaya ultrasonic classification standard. The difference of short-term outcome between the two groups were compared using t test, χ 2 test, Mann-Whitney U test, repeated measures design anova and multivariate analysis of variance. Results:At the 4th week after surgery, the VAS of mLSRS group was significantly lower than SB group(1 (2) vs. 2 (2) , F=5.32, P=0.02). One year after surgery, Constant-Murley score of two groups were significantly higher than those before surgery, but there was no significant difference between two groups(89.1±5.7 vs. 87.6±7.3, t=1.00, P=0.32). Abductor strength of affected limbs were lower than that of the opposite side, but there were no statistically significant differences between two groups(4.28±1.61 vs. 4.54±1.64, t=-0.70, P=0.49). According to the Sugaya classification, 32 cases in mLSRS group were healed while 7 cases were not healed,30 cases in SB group were healed while 9 cases were not healed(χ 2=0.32, P=0.58). Conclusion:The use of mLSRS fixation construct on large rotator cuff tear can reduce pain and the short-term outcome is reliable.

Objective:To examine the short-term outcomes of modified load-sharing rip-stop repair (mLSRS) and suture-bridge repair (SB) for large rotator cuff tear.Methods:The clinical data of patients who underwent shoulder arthroscopy for large rotator cuff repair at Department of Orthopedic, China Academy of Chinese Medical Sciences Wang Jing Hospital from January 2017 to December 2018 were collected. Totally 111 patients (39 in mLSRS group and 72 in SB group) were included in the study. There were 45 males and 66 females, aged (58.0±6.9)years (range: 40 to 72 years).A total of 39 pairs of patients were successfully matched by propensity score matching. The pain of shoulder joint was assessed by visual analogy score (VAS) , the shoulder function was assessed by Constant-Murley score, the abduction strength of bilateral shoulder joint was assessed by 90? force assessment method of upper limb on the plane of scapula, and the healing of rotator cuff on the operative side was assessed by sugaya ultrasonic classification standard. The difference of short-term outcome between the two groups were compared using t test, χ 2 test, Mann-Whitney U test, repeated measures design anova and multivariate analysis of variance. Results:At the 4th week after surgery, the VAS of mLSRS group was significantly lower than SB group(1 (2) vs. 2 (2) , F=5.32, P=0.02). One year after surgery, Constant-Murley score of two groups were significantly higher than those before surgery, but there was no significant difference between two groups(89.1±5.7 vs. 87.6±7.3, t=1.00, P=0.32). Abductor strength of affected limbs were lower than that of the opposite side, but there were no statistically significant differences between two groups(4.28±1.61 vs. 4.54±1.64, t=-0.70, P=0.49). According to the Sugaya classification, 32 cases in mLSRS group were healed while 7 cases were not healed,30 cases in SB group were healed while 9 cases were not healed(χ 2=0.32, P=0.58). Conclusion:The use of mLSRS fixation construct on large rotator cuff tear can reduce pain and the short-term outcome is reliable.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Objective: To investigate the differences of genotyping and virulence of hospital-acquired methicillin-resistant staphylococcus aureus (HA-MRSA) and community-acquired methicillin-resistant staphylococcus aureus (CA-MRSA). Methods: A total of 83 non-repetitive clinical specimens were collected in this study. All samples were classified into HA-MRSA and CA-MRSA according to the definition of the USA Centers for Disease Control and Prevention (CDC). Multiple polyemeras chain reaction (PCR) was used for SCCmec typing, and spa typing was conducted by means of PCR plus DNA sequencing. Panton-valentine leucocidin (PVL), phenol-soluble modulins α (PSMa), phenol-soluble modulins mec (PSM-mec), exfoliative toxin A (ETA), exfoliative toxin B (ETB), toxic shock syndrome toxin (TSST), α-toxin (hla), δ-toxin (hld) were also detected by conventional PCR. Results: The SCCmec type Ⅰ, Ⅱ, Ⅲ, Ⅳa and unclassified type accounted for 1.2%, 3.6%, 65.1%, 28.9% and 1.2%, respectively. 82.1% of the 39 strains of HA-MRSA were identified as SCCmec Ⅲ. While for the 44 strains of CA-MRSA, SCCmec Ⅲ and Ⅳa each accounted for 50%. The proportions of SCCmec Ⅲ and Ⅳa in HA-MRSA and CA-MRSA showed significant differences (χ²=9.343, 20.253, both P<0.05); There were 15 types of spa, t437, t062, t015 accounted for 39.8%, 21.7% and 10.8% respectively. Among the strains of HA-MRSA, the spa type t437 and t062 accounted for 28.2% and 23.1%, respectively, while for the spa type of CA-MRSA, t437 and t062 accounted for 50% and 20.5%, respectively. There was significant difference of t437 type between HA-MRSA and CA-MRSA (χ²=4.100, P=0.043). The positive rates of PSM-a, hla and hld were all 100%, and the positive rates of PVL, ETA, ETB, TSST and PSM-mec were 24.3%, 4.8%, 1.2%, 10.8% and 4.8%, respectively. The proportions of PVL positive HA-MRSA and CA-MRSA were 10.3% and 36.4%, respectively, which was significantly different (χ²=7.705, P=0.006), while PSM-mec positive HA-MRSA and CA-MRSA were 10.3% and 0%, respectively. Conclusions The strains of SCCmec Ⅳa and spa t437 are detected in HA-MRSA, suggesting that the more virulent CA-MRSA genotypes coexist with the traditional HA-MRSA genotypes in the hospital-acquired infections, which brings new challenges for the prevention and control of hospital infection.

Objective:To investigate the expression and clinical significance of PD-L1 in colorectal cancer (CRC). Methods:A total of 210 CRC patients who accepted radical surgery in our hospital from January 2015 to January 2016 were divided into three groups, namely, high-frequency microsatellite instability (MSI-H), low-frequency microsatellite instability (MSI-L), and microsatellite stable (MSS). The expression of PD-L1 was detected by immunohistochemistry, and the expression characteristics of PD-L1 in different types of CRC were analyzed. Results:CRC cases with low differentiation had a higher expression of PD-L1 than CRC patients with high differ-entiation (P<0.05). PD-L1 had a positive rate of 75.8%in the MSI-H group and a rate of 9.3%in the MSI-L and MSS groups, wherein the difference between the two groups was statistically significant (P<0.05). Conclusion:PD-L1 was positively expressed in some CRC tu-mor tissues, and its positive rate was significantly higher in MSI-H than in MSI-L and MSS. The therapeutic effect of a PD-L1 blocker for patients with MSI-H CRC might be preferable.