[摘 要] 目的：探究浸润性复层产生黏液的复层上皮癌（ISMC）的临床组织及分子病理特征。方法： 回顾性分析2018年1月至2024年4月间安徽医科大学安庆医学中心/安庆市立医院及皖南医学院第一附属医院/弋矶山医院的病理数据库的11例ISMC和4例产生黏液的复层上皮内病变（SMILE）的临床病理资料、免疫组化、阿利新蓝（AB）/过碘酸雪夫（PAS）染色、分子学检测及PD-L1表达情况。结果：ISMC患者多表现为阴道不规则流血。细胞质内含有黏液的细胞呈复层排列，周围呈栅栏状，肿瘤细胞可呈印戒样或胞质透明。ISMC不仅存在单纯型，也可呈混合型。ISMC具有高侵袭性的生物学特性。CK7、p16，p40和（或）p63表达呈癌巢周栅栏状阳性或局灶表达。AB/PAS染色阳性。人乳头状病毒（HPV）检测结果：ISMC中HPV16/18阳性（1/4），术前检测出HPV16/18阳性（4/6）；SMILE组织中HPV阴性。ISMC均表达PD-L1。成功随访8例ISMC患者，时间4~39个月（平均20.50月），4例SMILE患者，时间1~25个月（平均8.25月），随访患者均存活，1例ISMC术后出现多脏器转移。结论：ISMC具有独特的形态学特征及免疫表型，表现为高侵袭性和不良预后。所有ISMC均呈PD-L1阳性，提示所有患者均可从PD-L1免疫治疗中获益。

Objective To explore the mechanism of Medicoscab tincture in the treatment of second-degree burns based on network pharmacology and molecular docking technology. Methods The effective components of the tincture were screened by the TCMSP; the effective components of the tincture and burn related targets were screened by GeneCards and OMIM database; Cytoscape 3.7.2 software was used to draw “Chinese medicine-disease-effective components-targets” network diagram; the related gene ontology （GO） functions and pathways of the tincture were obtained through GO enrichment analysis and Kyoto encyclo-pedia of genes and geomes （KEGG） pathway analysis on the targets through Metascape platform; the pathway bubble diagram was constructed by the pathways enriched in the KEGG database, and finally verified by AutoDockTools for molecular alignment. Results 19 effective components and 179 target intersections of Medicoscab tincture were selected. GO analysis showed the intervention burns process mainly involved the reaction of inorganic substances, the reaction of cells to nitrogen compounds, and the response to xenobiotic stimuli, as well as biological processes such as membrane rafts, vesicular cavities, transcriptional regulatory factor complexes, receptor complexes, and endoplasmic reticulum cavities. KEGG analysis showed the function mainly includes AGE-RAGE signal pathway, PI3K-Akt signal pathway, IL-17 signal pathway, TNF signal pathway, etc. Analysis of cytoscape software showed the core targets were AKT1, TNF, IL-6, GAPDH, TP53, etc. Molecular docking showed that the active components of Medicoscab tincture were docking with multiple targets, among which β- sitosterol had strong binding activity with AKT1, GAPDH and TP53. Conclusion Quercetin, kaempferol, baicalein, β-sitosterol and other core active ingredients in the tincture of making scabs, which could assist in the relief of burns, regulate the signalling pathways such as AGE-RAGE, PI3K-Akt, IL-17, TNF and so on by acting on the targets of AKT1, GAPDH, TP53, IL-6 and so on. This study laid a theoretical foundation for clarifying the mechanism of action of tincture of scab making for the treatment of burn-like diseases.

Objective To compare the clinical application of empirical thoracoscopic segmentectomy and precise segmentectomy planned by artificial intelligence software, and to provide some reference for clinical segmentectomy. Methods A retrospective analysis was performed on the patients who underwent thoracoscopic segmentectomy in our department from 2019 to 2022. The patients receiving empirical thoracoscopic segmentectomy from January 2019 to September 2021 were selected as a group A, and the patients receiving precise segmentectomy from October 2021 to December 2022 were selected as a group B. The number of preoperative Hookwire positioning needle, proportion of patients meeting oncology criteria, surgical time, intraoperative blood loss, postoperative chest drainage time, postoperative hospital stay, and number of patients converted to thoracotomy between the two groups were compared. Results A total of 322 patients were collected. There were 158 patients in the group A, including 56 males and 102 females with a mean age of 56.86±8.82 years, and 164 patients in the group B, including 55 males and 109 females with a mean age of 56.69±9.05 years. All patients successfully underwent thoracoscopic segmentectomy, and patients whose resection margin did not meet the oncology criteria were further treated with extended resection or even lobectomy. There was no perioperative death. The number of positioning needles used for segmentectomy in the group A was more than that in the group B [47 (29.7%) vs. 9 (5.5%), P<0.001]. There was no statistical difference in the number of positioning needles used for wedge resection between the two groups during the same period (P=0.572). In the group A, the nodule could not be found in the resection target segment in 3 patients, and the resection margin was insufficient in 10 patients. While in the group B, the nodule could not be found in 1 patient, and the resection margin was insufficient in 3 patients. There was a statistical difference between the two groups [13 (8.2%) vs. 4 (2.4%), P=0.020]. There was no statistical difference between the two groups in terms of surgical time, intraoperative blood loss, duration of postoperative thoracic drainage, postoperative hospital stay, or conversion to open chest surgery (P＞0.05). Conclusion Preoperative surgical planning performed with the help of artificial intelligence software can effectively guide the completion of thoracoscopic anatomical segmentectomy. It can effectively ensure the resection margin of pulmonary nodules meeting the oncological requirements and significantly reduce the number of positioning needles of pulmonary nodules.

Seven triterpenoids were isolated and purified from the 95% aqueous EtOH extract whole plants of P. villosa by various chromatographic techniques, such as silica gel, ODS, Sephadex LH-20 gel column chromatography and preparative high performance liquid chromatography. Based on physicochemical properties and spectral analyses, the structures of the seven compounds were identified as 29-acetoxyoleanolic acid-3-O-α-L-arabinopyranoside (1), oleanolic acid (2), 3β-hydroxy-24-norursa-4(23),12,20(30)-trien-28-oic acid (3), 3β-hydroxy-24-nor-urs-4(23),12-dien-28-oic acid (4), ursolic acid (5), hederagenin (6), oleanolic acid 3-O-arabinoside (7). Compound 1 is a new compound. Compounds 3, 4, 6, and 7 are isolated from P. villosa for the first time. All compounds were assayed for their anti-inflammatory activity by the prodution of NO in lipopolysaccharide-stimulated RAW 264.7 cells. The results showed that compounds 1, 2, 3, 4, 6, and 7 significantly inhibited the NO release.

Objective To establish a regression equation for risk factors of refeeding syndrome (RFS) in critically ill elderly patients and analyze the relevant intervention measures. Methods Clinical materials of 154 critically ill elderly patients treated in Intensive Care Unit (ICU) from January 2021 to March 2023 were retrospectively analyzed, and they were divided into RFS group (n=51) and non-RFS group (n=103) according to incidence condition of RFS. The influencing factors of RFS were analyzed by Logistic regression model; the predictive values of predictors for RFS were analyzed by receiver operating characteristic (ROC) curve; the relevant Logistic regression equations were constructed and verified, and relevant nursing interventions were formulated. Results The incidence of RFS in critically ill elderly patients was correlated with the Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score, the Nutritional Risk Screening 2002 (NRS2002) score, invasive mechanical ventilation, fasting time before feeding, D-dimer level, nutritional intake methods, and pre-feeding blood phosphorus, potassium and magnesium levels (P < 0.05). Logistic regression analysis showed that APACHE Ⅱ score, NRS2002 score, nutrient intake methods and pre-feeding blood phosphorus and potassium levels were the independent risk factors of RFS in critically ill elderly patients (P < 0.05). The ROC curve results showed that the values of area under the curve (AUC) of APACHE Ⅱ score, NRS2002 score, nutritional intake methods, pre-feeding blood phosphorus and potassium levels, and the combined predictor in predicting the incidence of RFS in critically ill elderly patients were 0.754, 0.723, 0.707, 0.783, 0.774 and 0.859, respectively (P < 0.05). The Logistic regression equation for RFS was as follow: L=0.085×APACHE Ⅱ score-0.337×NRS 2002 score+0.537×nutrient intake methods-0.776×pre-feeding blood phosphorus level-0.207×pre-feeding blood potassium level+0.942. The predictive value of this equation was good, and targeted nursing interventions could be formulated based on this equation. Conclusion The regression equation of risk factors can be used for clinical prediction of the RFS risk in critically ill elderly patients, and clinical nursing interventions can be formulated based on the regression equation to prevent the occurrence of RFS.

This study aimed to analyze the impact of single nucleotide polymorphism (SNP) of ADCY3 (encoding adenylate cyclase 3) on the outcome of high-intensity interval training (HIIT) on body composition and screen genetic markers sensitive to HIIT in Chinese Han youth. A total of 237 non-regular exercise Han college students were recruited in a 12-week HIIT program, attending sessions 3 times a week. Before and after the HIIT program, their body composition was measured. DNA from the white blood cells was extracted and genotyped. PLINK (V1.09) software was used for quality control screening of SNPs loci, and a linear regression model was constructed to analyze the association between ADCY3 gene SNPs loci and body composition. ANOVA multiple comparisons (LSD) were performed to test the difference between groups, with the significance level set at 0.05. The results showed that: 1) A total of 22 SNPs loci were identified by the gene microarray scanning of ADCY3 gene, with 15 of them meeting the quality control criteria. The rs6753096 locus was associated with the training effect of HIIT on body composition; 2) The rs6753096 locus was not associated with pre-HIIT body composition; 3) Compared with volunteers with TT genotype, those with CT/CC genotype exhibited significant decrease in body mass index (BMI) and total body fat after training (P < 0.05); Male volunteers carrying the C allele had more significant training changes in skeletal muscle and lean body weight, while HIIT was more effective in decreasing body fat in female volunteers with CT/CC genotype; 4) The rs6753096 locus was significantly correlated with body fat sensitivity to HIIT (P = 0.0475), indicating that volunteers with CT/CC genotype were more sensitive to HIIT. In conclusion, 12-week HIIT program effectively improved the body composition of college students. The ADCY3 gene rs6753096 locus is not associated with pre-HIIT body composition, but it is associated with body composition sensitivity to HIIT, with individuals carrying CT/CC genotype showing greater responsiveness to HIIT.
Humans ; Adenylyl Cyclases/genetics* ; Male ; Female ; Body Composition/genetics* ; Polymorphism, Single Nucleotide ; Young Adult ; High-Intensity Interval Training/methods* ; Genotype ; Adult ; Adolescent

【Objective】 To explore the composition of culturable bacteria in platelets through bacterial culturomics and verify the results of culturomics and metagenomics to improve the detection rate of bacteria in platelets. 【Methods】 Platelet samples from 6 healthy people were collected. Eight kinds of culture media were placed in aerobic conditions and 12 kinds of culture media were placed in anaerobic conditions for large-scale culture and isolation of bacteria in platelets. The isolated single colony was identified by 16S rRNA gene sequencing. The bacterial abundance of healthy human platelet microbiome was analyzed by metagenomic sequencing, and the cultivable bacterial species in platelets was confirmed based on metagenomic and culturomics results. 【Results】 A total of 90 strains of bacteria belonging to 3 phylums, 5 classes, 5 orders, 7 families, 9 genus and 23 species were isolated from 6 platelet samples by culturomics. Among them, the strains with more monoclonal clones at the species level were Brevundimonas aurantiaca (16.7%), Bacillus sp. Y1 (15.6%), Cutibacterium acnes (14.4%) and Brevibacillus brevis (13.3%). The platelet samples sequenced by mNGS showed that the abundance values of Proteobacteria, Firmicutes and Actinobacteria were high. The bacteria detected by both culturomics and metagenomic sequencing methods were as follows: Firmicutes: Bacillus sp. Y1, B. thuringiensis, B. cereus, B. mobilis, B. velezensis, Staphylococcus epidermidis, and Brevibacillus brevis; Actinobacteria: Cutibacterium acnes; Proteobacteria: Escherichia coli and Delftia tsuruhatensis. 【Conclusion】 The mutual validation of culturomics and metagenomics has identified some bacteria, proving that bacteria exist in platelets.

【Objective】 To explore the influence of common methods of reducing non-viral nucleic acid on the abundance of plasma virus group. 【Methods】 Three kinds of library construction, five kinds of centrifugation conditions, two kinds of filters, four kinds of enzymes and four concentrations of chloroform were used to treat plasma samples added quantitatively 2.16 mL of pseudorabies virus(PRV) and 2.16 mL of porcine parvovirus(PPV). A total of 21.6 mL of plasma samples were processed, including 54 samples. Subsequently, nucleic acid was extracted, mitochondrial DNA(mtDNA) and two viruses were quantitated, the library of the next generation sequencing was constructed, Illumina NovaSeq 6000 was used for the next generation sequencing. The sequencing data were compared with Kraken Py 2.0 software, and the species annotation analysis was conducted. The corresponding species classification information of each segment was obtained to analyze the impact of different reducing non-viral nucleic acid methods on the relative abundance of microorganisms and two indicator viruses. 【Results】 After sequencing by Illumina NovaSeq 6000, 306.27 GB raw data and 193.17 GB clean data were obtained, with Q20>90%, Q30>85%, Error Rate of 0.03%, and average GC Content of 45.02%. The DNA library construction process significantly increased the proportion of microbial sequences and the PRV abundance [(91.8±0.5)%](P<0.05); RNA library construction and combined library construction can increase the abundance of Pestivirus, an RNA virus, and the PRV abundance was(17.7±3.3)% and(8.1±1.5)% respectively. The Ct value of mtDNA was increased and the proportion of human sequence decreased to less than(89.5±1)%, while the proportion of microbial sequence increased to (2.4±0.03)% after treatment of five centrifugation conditions(P<0.05); After centrifugation at 4℃, 100 g, 30 min, the PRV abundance was increased to (40.6±6)%, and centrifugation at 4℃, 4 000 g, 45 min reduced the PRV abundance to (4.1±0.01)%(P<0.05). Both of 0.22-μm filter and 0.45-μm filter increased the Ct value of mtDNA to above 25.56±0.13, decreased the proportion of human sequence to less than (86.1±0.6)%, increased the proportion of microbial sequence to (3.1±0.1)% and (3.4±0.2)%, and decreased the PRV abundance to (1.6±0.3)% and (4.1±0.7)%(P<0.05), while there was no statistical difference in the effect on PPV concentration and abundance. DNase Ⅰ and Benzonase increased the Ct value of PPV to 25.65±0.06 and 25.36±0.45, decreased the proportion of human sequence to (81.7±5.6)% and (72.8±6.7)%, and increased the proportion of microbial sequence and PRV abundance to (11.0±4.1)% and (16.1±4.7)%, (55.8±2.3)% and (39.0±8.9)%, respectively(P<0 05); After treatment with RNase A, the Ct value of PRV increased to 25.20±0.11, and the human sequence proportion decreased to (85.4±5.6)%(P<0 05); Lysozyme had no effect on removing non-viral nucleic acid. The chloroform of 1%, 5%, 10% and 20% increased Ct value of PRV and mtDNA to no less than 27.17±0.21 and 25.68±0.04; Only 10% chloroform increased the proportion of microbial sequences to (3.1±1.2)%(P<0.05); The abundance of PRV with 1% and 5% chloroform treatment was increased to (48.7±13.3)% and (42.1±5.5)%(P<0.05), while 10% and 20% chloroform reduced PRV abundance to (1.0±0.5)% and (3.4±2.8)%(P<0.05). There was no statistical difference in the effect of chloroform with four contents on PPV abundance. 【Conclusion】 Centrifugation at 4℃, 5 000 g, 10 min is suitable for increasing the overall abundance of virus, and centrifugation at 4℃, 100 g, 30 min is suitable for increasing the content of virus similar to PRV. 0.45-μm filter, DNase Ⅰ, Benzonase and low concentration chloroform can effectively reduce the proportion of non-viral nucleic acid sequence in plasma to increase the abundance of the indicated virus group. Thus, the enrichment effect of plasma meta-virome is closely related to the nature of the virus, and the appropriate virus enrichment method should be selected according to the research purpose to establish the corresponding enrichment strategy.

【Objective】 To evaluate the application value of nucleic acid testing (NAT) by studying the NAT-yield of syphilis screening reactive blood from five blood centers. 【Methods】 The blood samples and demographic information of syphilis screening positive donors were collected from five domestic blood centers, i. e. Chongqing, Guangxi, Luoyang, Liuzhou, Mianyang and Urumqi. The treponema pallidum particle agglutination (TPPA) and the established SYBR Green qPCR method were used to analyze the difference between the results of NAT and the other two test results. 【Results】 Among 1 679 reactive blood samples for syphilis screening, 819 were confirmed positive by TPPA, accounting for 49%, with the false positive rate exceeded 50%. As to NAT results, the NAT-yield of syphilis screening reactive samples and confirmed positive samples was the same (both 2.20%); the NAT-yield of TPPA-positive and TPPA-negative samples were 2.20% and 2.74%, respectively. 【Conclusion】 Primary syphilis screening by ELISA has high sensitivity, but also presents high false positive rate. Although TPPA confirmatory test has strong specificity, it cannot reflect the existence of T. pallidum. Therefore, NAT may be used as a supplementary test for syphilis screening so as to more effectively ensure the safety of blood transfusion and blood supply.

{L-End}Objective To explore the feasibility of using positron emission tomography (PET) -computed tomography (CT) to detect brain metabolic abnormalities caused by trimethyltin chloride (TMT) poisoning. {L-End}Methods Specific pathogen free healthy SD rats were randomly divided into model group and control group with six rats in each group. Rats in the model group were intraperitoneally injected with a single dose of 10 mg/kg body mass of TMT solution, and rats in the control group were intraperitoneally injected with a single dose of an equal volume of 0.9% sodium chloride solution. Rats were anaesthetized after three days of modeling and underwent PET-CT brain scanning to detect the standardized uptake value (SUV) of ¹⁸F-2-fluro-D-deoxy-glucose (¹⁸F-FDG). After scanning, rats were sacrificed and brain tissues were collected for brain organ coefficients calculation and brain histopathological analysis. {L-End}Results The rats in the model group showed symptoms of head tremor, limb twitching, irritability and others after TMT modeling. There was no significant difference in the body mass between the two groups of rats on the third day of modeling (P>0.05). The ¹⁸F-FDG uptake in the cerebral cortex, cerebellum and brainstem of the rats in the model group was significantly weakened compared with the control group, with deceased SUV values (all P<0.05). No obvious abnormalities were found in CT images and freshly collected brain tissues of rats of the control and model groups. The brain organ coefficients of rats in the two groups showed no significant difference (P>0.05). The results of hematoxylin-eosin staining of brain tissue showed that the cerebral cortex of rats in the model group had more tiny cavities than that of the control group, and some neuronal cells and a small number of hippocampal vertebral cells were tightly and deeply stained, with the cytoplasm and nucleus poorly demarcated, and pericellular space enlarged. The results of Nissen staining showed that the arrangement of neuronal cells in the model group was slightly disordered, and the interstitial space was slightly enlarged, but no other significant abnormal changes were observed. {L-End}Conclusion PET-CT can be used in detecting the metabolic abnormalities of brain in TMT poisoning rat model, making it a sensitive detection method for TMT poisoning.