Objective To establish a sensitive,accurate and rapid detection method for 10 drugs and metabolites in hair with triple quadrupole tandem mass spectrometry,addressing the identification of drugs in real hair samples.Methods After cryogenic grinding and ultrasonic extraction,hair was separated by Restek Allure PFPP column(100 mm × 2.1 mm,5 μm).The mobile phase A was 20 mmol/L ammonium acetate,0.1％formic acid and 5％acetonitrile aqueous solution.The mobile phase B was acetonitrile.An electrospray ionization source was used for data acquisition in scheduled MRM mode.Results The method showed good linearity for all analytes within the validated range(R2＞0.995),with the limits of detection ranging from 0.5 to 6 pg/mg,the limits of quantification ranging from 2.5 to 10 pg/mg.Accuracy ranged from 89.1％to 114.6％,with intra-day precision ranging from 0.2％to 11.7％,inter-day precision ranging from 4.0％to 15.8％.the matrix effects ranging from 89.4％to 118.4％,the recoveries ranging from 63.6％～112.1％.Totally 20 cases were detected positive in 196 actual hair samples.Conclusion The time-scheduled scanning method exhibits high stability and sensitivity,enabling high-throughput detection,and is suitable for forensic toxicology laboratories to identify drugs in hair.

Objective To establish a sensitive,accurate and rapid detection method for 10 drugs and metabolites in hair with triple quadrupole tandem mass spectrometry,addressing the identification of drugs in real hair samples.Methods After cryogenic grinding and ultrasonic extraction,hair was separated by Restek Allure PFPP column(100 mm × 2.1 mm,5 μm).The mobile phase A was 20 mmol/L ammonium acetate,0.1％formic acid and 5％acetonitrile aqueous solution.The mobile phase B was acetonitrile.An electrospray ionization source was used for data acquisition in scheduled MRM mode.Results The method showed good linearity for all analytes within the validated range(R2＞0.995),with the limits of detection ranging from 0.5 to 6 pg/mg,the limits of quantification ranging from 2.5 to 10 pg/mg.Accuracy ranged from 89.1％to 114.6％,with intra-day precision ranging from 0.2％to 11.7％,inter-day precision ranging from 4.0％to 15.8％.the matrix effects ranging from 89.4％to 118.4％,the recoveries ranging from 63.6％～112.1％.Totally 20 cases were detected positive in 196 actual hair samples.Conclusion The time-scheduled scanning method exhibits high stability and sensitivity,enabling high-throughput detection,and is suitable for forensic toxicology laboratories to identify drugs in hair.

Objective:To compare the clinical outcomes of arthroscopic external tension band fixation versus open reduction and internal fixation in the treatment of greater tubercle fracture of the humerus.Methods:A retrospective cohort study was conducted on 55 patients with greater tubercle fracture of the humerus admitted to Taizhou Hospital of Zhejiang Province from September 2019 to June 2022, including 24 males and 31 females, aged 26-80 years [(61.7±10.5)years]. Out of them, 35 patients treated with open reduction and internal fixation (open reduction group), and 20 patients were treated with external anchor tension band under arthroscopy (arthroscopy group). The operation time, and the Visual Analogue Scale (VAS) score, American Shoulder and Elbow Surgeons (ASES) score, Constant-Murley score and shoulder active range of motion (anterior flexion, abduction and posterior extension) before operation, at 1 month after operation and at the last follow-up were compared between the two groups. Bone healing was observed in both groups at the last follow-up. Postoperative complications were compared between the two groups.Results:All the patients were followed up for 12-29 months [(16.9±4.0)months]. There was no significant difference in operation time between the two groups ( P>0.05). There were no significant differences in the VAS score, ASES score, Constant-Murley score and shoulder active range of motion between the two groups before operation ( P>0.05). The VAS score of the arthroscopy group was 3(2, 3)points at 1 month after operation, which was significantly lower than that of the open reduction group [4(3, 4) points] ( P<0.01). No significant difference was found in the VAS score at the last follow-up between the two groups ( P>0.05).The ASES scores of the arthroscopy group were (70.6±4.2)points and (90.2±3.7)points at 1 month after operation and at the last follow-up respectively, which were significantly higher than those of the open reduction group [(64.7±6.4)points and (87.5±4.9)points respectively] ( P<0.05 or 0.01). There was no significant difference in the Constant-Murley score between the arthroscopy group [(71.8±4.3)points] and the open reduction group [(70.9±5.3)points] at 1 month after operation ( P>0.05), while the Constant-Murley score of the arthroscopy group was (94.1±3.1)points at the last follow-up, which was significantly higher than that of the open reduction group [(89.2±4.7)points] ( P<0.01). At 1 month after operation and at the last follow-up, ranges of motion of the anterior flexion, abduction and posterior extension were (52.7±12.3)° and (140.0±16.9)°, (57.4±8.6)° and (125.0±14.3)°, and 16(15, 19)° and 25(20, 30)° in the arthroscopy group respectively, which were significantly higher than those in the open reduction group [(42.2±5.2)° and (110.9±14.0)°, (52.8±6.0)° and (103.7±11.7)°, and 10(10, 20)° and 16(15, 25)° respectively] ( P<0.05 or 0.01). At the last follow-up, it was found that bony union was achieved in both groups. There were no obvious complications such as incision infection or joint stiffnessin both groups. In the open reduction group, 2 patients had internal fixation failure within 1-3 months after operation but was treated with revision operation; 6 patients developed shoulder stiffness at 3-6 months after operation but had outpatient rehabilitation. The incidence rate of postoperative complications in the arthroscopy group [0%(0/20)] was significantly lower than that in the open reduction group [23%(8/35)] ( P<0.05). Conclusion:Compared with open reduction and internal fixation with plates and screws, arthroscopic external anchor tension band fixation in the treatment of greater tuberosity fracture of the humerus has the advantages of earlier pain relief, better shoulder functional improvement, better recovery of shoulder mobility, and fewer complications.

In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.

Objective To detect and analyze the in vitro metabolites of amfepramone using a rat liver microsomal model by liquid chromatography coupled with high-resolution tandem mass spectrometry(LC-HR-MS/MS),and compare them with those derived from authentic urine samples collected from amfepramone users,so as to evaluate the consistency of the in vitro rat liver microsomal model in predicting the metabolites in vivo.Methods Wistar rat liver microsomes were exposed to 5 μg amfepramone and subsequently incubated for 1 h to simulate the metabolic process in the human body.The authentic urine samples collected from amfepramone users were precipitated with acetonitrile.Results A total of of ten phase I metabolites of amfepramone were produced in the in vitro rat liver microsome model.Eight out of the ten in vitro phase I metabolites were also found in authentic urine samples collected from amfepramone users.The primary metabolic pathways were deduced to be carbonyl reduction,N-dealkylation,and hydroxylation.This in vitro metabolic model-based approach proved to be a simple and rapid method for predicting amfepramone metabolites in vivo.However,it was observed that certain metabolites of amfepramone,including M2(ethcathinone)and M4(cathinone),possess non-exclusive origins,limiting their suitability as standalone biomarkers for detecting amfepramone use in urine specimens.Conclusion This study revealed the involvement of the hydroxylation pathway in amfepramone metabolism.Additionally,four new metabolites were identified in real urine samples from amfepramone users.These preliminary results contribute valuable insights into the pharmacokinetics and forensic analysis of amfepramone,providing a theoretical foundation for further research in these domains.

Objective To detect and analyze the in vitro metabolites of amfepramone using a rat liver microsomal model by liquid chromatography coupled with high-resolution tandem mass spectrometry(LC-HR-MS/MS),and compare them with those derived from authentic urine samples collected from amfepramone users,so as to evaluate the consistency of the in vitro rat liver microsomal model in predicting the metabolites in vivo.Methods Wistar rat liver microsomes were exposed to 5 μg amfepramone and subsequently incubated for 1 h to simulate the metabolic process in the human body.The authentic urine samples collected from amfepramone users were precipitated with acetonitrile.Results A total of of ten phase I metabolites of amfepramone were produced in the in vitro rat liver microsome model.Eight out of the ten in vitro phase I metabolites were also found in authentic urine samples collected from amfepramone users.The primary metabolic pathways were deduced to be carbonyl reduction,N-dealkylation,and hydroxylation.This in vitro metabolic model-based approach proved to be a simple and rapid method for predicting amfepramone metabolites in vivo.However,it was observed that certain metabolites of amfepramone,including M2(ethcathinone)and M4(cathinone),possess non-exclusive origins,limiting their suitability as standalone biomarkers for detecting amfepramone use in urine specimens.Conclusion This study revealed the involvement of the hydroxylation pathway in amfepramone metabolism.Additionally,four new metabolites were identified in real urine samples from amfepramone users.These preliminary results contribute valuable insights into the pharmacokinetics and forensic analysis of amfepramone,providing a theoretical foundation for further research in these domains.

Objective:To confirm the mechanism of Sedum alfredii extract (SafE) alleviating radiation injury in human small intestinal epithelial cells (HIEC-6). Methods:HIEC-6 cells were divided into 4 groups, including control group (Con), irradiation group (IR), SafE alone group (SafE) and SafE plus irradiation group (SafE+ IR). All of the SafE groups were treated with 0.02 g/ml (W/V) SafE for 24 h. Cell viability (CCK-8 method ) and intracellular ROS levels were investigated at 24 h after 2, 4, and 6 Gy irradiation. Samples were taken at 24 h after 4 Gy irradiation for transcriptome analysis, and the intracellular E3 ubiquitin ligase PRKN expression level was measured. The thickness of endoplasmic reticulum was detected at 24 h after 4 Gy irradiation using fluorescent dye.Results:SafE could maintain cell viability after irradiation ( t=2.94-10.40, P<0.05), and significantly reduced the level of ROS in the irradiated cells ( t=-13.29--4.53, P<0.05). PRKN was preliminarily verified to be the target gene of SafE that maintained PRKN transcript level and endoplasmic reticulum thickness after irradiation (IR group vs. Con group: t=-5.55, 3.27, P<0.05, SafE group vs. SafE+ IR group: P>0.05). Conclusion:SafE is effective in maintaining ER thickness and reducing cellular radiation damage and its target gene PRKN could be regulated by ionizing radiation.

Photoimmunotherapy (PIT) is an emerging tumor-targeted phototherapy that combines the tumor specificity of monoclonal antibodies with the phototoxicity of light absorbers to rapidly and selectively induce the immunogenic death of target tumor cells. PIT has minimal side effects due to its high specificity. The immunogenic cell death induced by PIT results in rapid maturation of immature dendritic cells proximal to dying tumor cells. Subsequently, the mature dendritic cells present the tumor antigens to CD8+ T cells and induce their activation and proliferation, thus enhancing the antitumor immune response of the host. PIT can also improve the anticancer efficacy by enhancing the penetration of nanomedicines into tumor tissues. In view of the excellent application prospects of PIT, this review summarizes the advances in the immune activation mechanism of PIT, the superenhanced permeability and retention effect, and the new strategies for combinatory therapy, providing references for future research and clinical translation.
Antibodies, Monoclonal/therapeutic use* ; Humans ; Immunotherapy ; Neoplasms/therapy* ; Photosensitizing Agents ; Phototherapy

Near infrared photoimmunotherapy (NIR-PIT) is a highly selective molecularly targeted phototherapy for cancer which is based on injecting a conjugate of IRDye700DX,a water-soluble near-infrared silicon-phthalocyanine dye,and a monoclonal antibody that targets an antigen on the cancer cell surface. Subsequent local irradiation of NIR light causes the rapid and specific tumor cell death. Due to the good clinical translation prospects of NIR-PIT,this paper summarizes the influencing factors,antitumor mechanism,main challenges and recent strategies,which may benefit for its research and clinical application.

The recurrence of colorectal polyps is caused by various factors and leads to the carcinogenesis of colorectal cancer, which ranks third in incidence and fourth in mortality among cancers worldwide. The potential risk factors for colorectal polyp recurrence have been demonstrated in multiple trials. However, an article that pools and summarizes the various results is needed. This review enumerates and analyzes some risk factors in terms of patient characteristics, procedural operations, polyp characteristics, and dietary aspects to propose some effective prophylactic measures. This review aimed to provide a reference for clinical application and guide patients to prevent colorectal polyp recurrence in a more effective manner.