Objective To establish a rapid and convenient UPLC-MS/MS method for the determination of tamoxifen and its active metabolite endoxifen in plasma of patients with breast cancer.Methods With diazepam as the internal standard,the plasma sample was precipitated with acetonitrile and methanol-acetonitrile was used as mobile phase.The samples were separated by chromatographic column ZORBAX Eclipse plus C18(18 μm,2.1 mm x50 mm)and scanned by electrospray ion source and positive ion multiple reaction monitoring mode.The ion channels were detected as tamoxifen m/z 372.0→72.1,endoxifen m/z 374.0→58.2 and diazepam m/z 285→192.9.Results The linearity of tamoxifen and endoxifen were good in the concentration range of 1-500 ng·mL-1(R2=0.999 5)and 0.5-250 ng·mL-1,respectively(R2=0.999 9).The intra-day and inter-day precision of low,medium and high quality control concentrations were all less than 10％.Tamoxifen and endoxifen were stable at low temperature and stable after repeated freeze-thaw.Conclusion The experimental results show that the method meets the requirements of biological sample analysis,and the sample treatment is simple,specific,and can be used for the accurate and rapid determination of tamoxifen and indoloxifen in human plasma.

Diabetic kidney disease(DKD)is one of the most important complications of diabetes.In recent years,domestic and foreign studies have found that mammalian target protein of rapamycin(mTOR)related signaling pathway is a classic pathway involved in the regulation of autophagy,which can achieve the therapeutic effect of DKD by targeting the autophagy pathway,and plays a crucial role in the prevention and treatment of DKD.In this paper,we reviewed the mechanism of mTOR-related signaling pathway targeted autophagy in the prevention and treatment of DKD,in order to provide a new reference and basis for clinical prevention and treatment of DKD.

Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

Objective To overcome the limitations of existing human respiratory syncytial virus(hRSV)animal models,such as semi-permissiveness and short duration of infection,this study established an IL2rg gene knockout(IL2rg-/-)rat model using TALEN gene editing technology.Methods The animal model was infected with hRSV intranasally.Clinical characteristics,body weight,and temperature changes were observed over the infection period(0～35 days).The total viral loads in respiratory organs,such as the nasal tissue,trachea,and lungs,were measured at various time points(4,11,20,and 35 days post-infection).Pathological analysis was conducted on target organs at the endpoint of observation(35 days post-infection).Changes in peripheral blood T,B,NK,and NKT cells and various cytokines were assessed at various time points(4,20,and 35 days post-infection).Results(1)IL2rg/-knockout rats sustained high viral loads in the nasal cavity upon intranasal inoculation with hRSV.The average peak titer rapidly reached 1 × 1010 copies/g in nasal tissue and 1 × 107 copies/g up to 5 weeks post-infection.(2)However,no significant pathological changes were noted in nasal,tracheal,or lung tissues.(3)An increase was observed in the content of peripheral blood B cells in hRSV-infected IL2rg--rats.(4)IL-6 and MCP-1 were increased in the early stage of infection and then decreased at the end of the observation period.Conclusions This study established a new IL2rg-/-rat model using TALEN technology and found that this model effectively supported high-level replication and long-term infection of hRSV,providing a good basis for antiviral drug screening and in vivo efficacy evaluation of anti-hRSV antibodies.

The steps for installation and withdrawal of WYD2000 field surgical lamp were introduced.The failures and causes of broken cross-arm connector of WYD2000 field surgical lamp were analyzed.The problems of WYD2000 field surgical lamp in vulnerability to breaking and difficulty in maintenance were solved by designing and manufacturing a special maintenance tool and optimizing the materials and fixing mode of cross-arm connection.References were provided for main-tenance and improvement of WYD2000 field surgical lamp.[Chinese Medical Equipment Journal,2024,45(4):116-118]

Objective:To explore the characteristics of drug-induced liver injury induced by acetaminophen (APAP) in children, and to clarify the mechanism of improving APAP liver injury in children on targeting MCJ.Methods:The C57BL/6 young and adult mice and mouse liver parenchymal AML-12 cells were used in the prospective study. Forty C57BL/6 young mice of 3 to 4 weeks and 40 C57BL/6 adult mice of 6 to 8 weeks were randomly divided into 5 groups (control 0 h group, APAP 3 h group, APAP 6 h group, APAP 12 h group, APAP 24 h group), 8 rats in each group and intraperitoneally injected with APAP (300 mg/kg) to replicate drug-induced liver injury (DILI) model. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected. Hematoxylin-eosin staining (H&E) staining was used to observe the pathological change of liver tissue. The oxidative damage indexes lactate dehydrogenase (LDH), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) of liver tissue homogenate were detected. The expression of MCJ protein was detected by Western blot. Mouse liver parenchymal AML-12 cells were transfected with MCJ interference sequence and APAP to construct hepatocyte injury model. Hepatocyte viability, LDH and SOD levels were measured.Results:As shown in the animals results, after 3 h of APAP administration, ALT and AST levels were increased significantly in young mice, compared with adult mice: (105.25 ± 18.38) U/L vs. (37.38 ± 15.75) U/L, (87.38 ± 30.68) U/L vs. (28.50 ± 19.44)U/L, which suggested that the time liver necrosis occurred in young mice was earlier than that of adult mice. Moreover, the levels of LDH and MDA were significantly increased, while GSH and SOD were significantly decreased after 3 h of APAP in young mice, compared with adult mice, (1 231.38 ± 183.94) U/L vs. (810.50 ± 190.64) U/L, (19.63 ± 3.58) nmol/mg vs. (16.50 ± 3.51) nmol/mg, (39.75 ± 4.62) nmol/mg vs. (53.63 ± 13.07) nmol/mg, (51.38 ± 6.37)% vs. (63.25 ± 9.13)% ( P<0.01), and the levels of MCJ protein were significantly increased. After MCJ knockdown, hepatocyte viability and LDH, SOD levels of mouse liver parenchymal AML-12 cells were significantly improved compared with APAP model group ( P<0.01). Conclusions:APAP liver injury was more common in young mice than in adult mice. Knockdown MCJ expression may ameliorate APAP liver injury in young mice. MCJ may serve as a target for early intervention of APAP liver injury in children, which provide new therapeutic targets and strategies for APAP liver injury in children.

Chronic kidney disease(CKD)commonly used dietary assessments including 24-hour dietary recall(24 h DR)/3-day dietary recall(3DDR),food frequency questionnaire(FFQ),dietary records,and estimation of dietary protein intake based on nitrogen balance.Given the high prevalence of CKD patients in Asian population and the scarcity of research using FFQ method,it is crucial to develop an FFQ suitable for Chinese CKD patients.This review summarizes the advantages and disadvantages of dietary assessment methods for CKD,the current research status,and the content and steps involved in establishing an FFQ,with the aim of providing reference for the modification of FFQ for Chinese CKD patients.

Objective To investigate the effect of rapid ultrasonic tissue processing on breast biopsy tissue.Methods Totally 100 breast puncture samples from the Department of Pathology at Shengjing Hospital of China Medical University in 2023 were randomly selected and dehydrated using rapid ultrasound tissue processing technology(the rapid group).Paired surgical samples(0.3 cm×0.3 cm×0.3 cm)that did not undergo neoadjuvant therapy before surgery were collected and subjected to routine pathological tissue treat-ment(the conventional group).The two groups of samples were subjected to routine sectioning after dehydration.The dehydration time,hematoxylin and eosin(HE)staining effect,and fluorescence in situ hybridization(FISH)results of the two groups were compared.Next,the estrogen receptor(ER),progesterone receptor(PR),and human epidermal growth factor receptor 2(HER2)immunohistochemical(IHC)detection results of the two groups were compared using the kappa test with paired counting data.Results Dehydration time in the rapid group(46.70±1.42 min)was significantly shorter than that in the conventional group(659.25±6.03 min,t=987.44,P<0.001).No significant differences were observed between the HE staining scores of the rapid group(10.13±1.12)and that of the conventional group(10.15±0.97).The consistency rate of ER expression between the two groups was 92.5%(Kappa=0.794,P<0.05),that of PR expres-sion between the two groups was 76.25%(Kappa=0.639,P<0.05),and that of HER2 expression between the two groups was 72.5%(Kappa=0.610,P<0.05).The FISH results were consistent between the two groups.Conclusion Rapid ultrasonic tissue treatment has the advantages of a short procedure time,less pollution,and simple operation.The present study showed no statistically significant differences between the effect of rapid tissue treatment on tissue sections and that of routine dehydration.Therefore,this method can be widely used in pathology for the analysis of small samples.

Objective To observe the alterations in functional complexity of brain regions in autism spectrum disorder(ASD)patients and correlations with the predicted brain age.Methods Open brain resting-state functional MRI(rs-MRI)data of 93 ASD patients and 96 typically developing adolescents(healthy subjects)were downloaded.The functional complexity in brain regions were extracted with self-developed virtual digital brain software,and the alterations in functional complexity of brain regions in ASD patients and correlations with their ages were analyzed.Two networks were prospectively trained with data of 65 ASD patients and 67 healthy subjects as the training set to predict brain age,and the results were evaluated,and the predicting errors were compared using test set,i.e.the other 28 ASD patients and 29 healthy subjects.Results Compared to healthy subjects,on the basis of anatomical automatic labeling(AAL)atlas,ASD patients exhibited significantly reduced functional complexity based on Shannon entropy in the left precuneus,left cuneus and right parahippocampal gyrus.Conversely,functional complexity of ASD patients based on permutation entropy significantly increased in the left cuneus and right cerebellar Crus Ⅱ region.The left hippocampus showed reduced functional complexity based on Pearson correlation coefficient,while the left middle temporal gyrus showed increased functional complexity based on Pearson correlation coefficient.The functional complexity in brain regions of ASD patients were not closely correlated with ages(all|r|＜0.4).According to the trained fully connected network,the predicted brain ages of ASD patients and healthy subjects in test set were all lower than their physiological ages,but no significant difference was found between the prediction errors of ASD patients and healthy subjects(P=0.283).Conclusion Functional complexity changed in some brain region functions in ASD patients.The predicted brain ages of ASD patients based on the obtained fully connected network were on the low side,but not obviously affected by the alterations of functional complexity in brain regions.

Objective:To explore the mechanism of herb-partitioned moxibustion in relieving rat intestinal inflammation by focusing on the neutrophil extracellular traps(NETs)in Crohn disease(CD)development. Methods:Rats were randomly divided into a normal group,a model group,a herb-partitioned moxibustion group,and a mesalazine group.The CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid except for rats in the normal group.Rats in the normal group and model group did not receive any treatment but had the same fixation as the other groups.Rats in the herb-partitioned moxibustion group received herb-partitioned moxibustion at Qihai(CV6)and bilateral Tianshu(ST25).Rats in the mesalazine group received intragastric administration of mesalazine enteric-coated tablets.The general situation of rats in each group was recorded,and the histopathological changes in the colon were observed and scored by hematoxylin-eosin staining.The serum concentrations of NETs DNA(NETs-DNA),neutrophil elastase(NE)-DNA,and myeloperoxidase(MPO)-DNA were detected by ABC enzyme-linked immunosorbent assay,and the citrullinated histone 3(citH3),MPO,and NE protein and mRNA expression levels in rat colon tissue were observed by immunofluorescence and real-time quantitative polymerase chain reaction. Results:Compared with the normal group,the mucosal ulcer reached the muscularis,the epithelium was incomplete,the goblet cells decreased obviously with significant inflammatory cell infiltration in the colon;the colonic mucosa damage index(CMDI)score increased significantly(P<0.01);the serum NETs-DNA,NE-DNA,and MPO-DNA concentrations increased(P<0.05);the NE,citH3,and MPO protein and mRNA expression in the colonic tissue increased significantly in the model group(P<0.01 or P<0.05).Compared with the model group,the mucosal epithelium in the herb-partitioned moxibustion group and the mesalazine group was repaired and the goblet cells increased with a few infiltrating inflammatory cells in the colon;the CMDI score decreased(P<0.01);the serum NETs-DNA,NE-DNA,and MPO-DNA concentrations decreased(P<0.05);the NE,citH3,and MPO protein and mRNA expression in the colonic tissue was down-regulated(P<0.01 or P<0.05). Conclusion:Herb-partitioned moxibustion reduced the serum NETs complex and inhibited the protein and mRNA expression of NETs complex in the colon tissue,which may be one mechanism of herb-partitioned moxibustion in relieving colon mucosal inflammation in CD.