As one of the key enzymes in cell metabolism, the activity of citrate synthase 3 (CS3) regulates the substance and energy metabolism of organisms. The protein members of CS3 family were identified from the whole genome of apple, and bioinformatics analysis was performed and expression patterns were analyzed to provide a theoretical basis for studying the potential function of CS3 gene in apple. BLASTp was used to identify members of the apple CS3 family based on the GDR database, and the basic information of CS3 protein sequence, subcellular localization, domain composition, phylogenetic relationship and chromosome localization were analyzed by Pfam, SMART, MEGA5.0, clustalx.exe, ExPASy Proteomics Server, MEGAX, SOPMA, MEME, WoLF PSORT and other software. The tissue expression and inducible expression characteristics of 6 CS3 genes in apple were determined by acid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Apple CS3 gene family contains 6 members, and these CS3 proteins contain 473-608 amino acid residues, with isoelectric point distribution between 7.21 and 8.82. Subcellular localization results showed that CS3 protein was located in mitochondria and chloroplasts, respectively. Phylogenetic analysis divided them into 3 categories, and the number of genes in each subfamily was 2. Chromosome localization analysis showed that CS3 gene was distributed on different chromosomes of apple. The secondary structure of protein is mainly α-helix, followed by random curling, and the proportion of β-angle is the smallest. The 6 members were all expressed in different apple tissues. The overall expression trend from high to low was the highest relative expression content of MdCS3.4, followed by MdCS3.6, and the relative expression level of other members was in the order of MdCS3.3 > MdCS3.2 > MdCS3.1 > MdCS3.5. qRT-PCR results showed that MdCS3.1 and MdCS3.3 genes had the highest relative expression in the pulp of 'Chengji No. 1' with low acid content, and MdCS3.2 and MdCS3.3 genes in the pulp of 'Asda' with higher acid content had the highest relative expression. Therefore, in this study, the relative expression of CS3 gene in apple cultivars with different acid content in different apple varieties was detected, and its role in apple fruit acid synthesis was analyzed. The experimental results showed that the relative expression of CS3 gene in different apple varieties was different, which provided a reference for the subsequent study of the quality formation mechanism of apple.
Citric Acid ; Malus/genetics* ; Citrate (si)-Synthase ; Phylogeny ; Citrates

Objective:To investigate the application value of major anatomical structure recognition model of minimally invasive liver resection based on deep learning.Methods:The retrospective and descriptive study was conducted. The 31 surgical videos of laparoscopic left lateral sectionectomy performed in Zhujiang Hospital of Southern Medical University from January 2019 to April 2023 were collected. Video clips containing the surgical procedure of left lateral lobe liver pedicle and left hepatic vein were screened by 2 liver surgeons. After quality control, screening and frame extraction, the major anatomical structures on the images of these clips were annotated. After pre-processing, these images were transported to the DeepLab v3+neural network framework for model training. Observation indicators: (1) video annotation and classification; (2) results of arti-ficial intelligence anatomical recognition model testing. Measurement data with normal distribution were represented as Mean± SD, and count data were described as absolute numbers. Results:(1) Video annotation and classification. A total of 4 130 frames of images were annotated in the 31 surgical videos, including 2 083 frames of annotated images for the left lateral lobe liver pedicle, 1 578 frames of annotated images for the left hepatic vein and 469 frames of annotated images for both the left lateral lobe liver pedicle and left hepatic vein. (2) Results of artificial intelligence anatomical recognition model testing. In four application scenarios (clean scene, bloodstain scene, partially obstruction by instrument scene, and small exposed area scene), the model was able to successfully recognize the left lateral lobe liver pedicle and left hepatic vein, with a recognition speed for anatomical markers >13 frames/s. When performing anatomical recognition on images with only the left lateral lobe liver pedicle, the Dice coefficient, intersection over union, accuracy, sensitivity and specificity of the model were 0.710±0.110, 0.560±0.120, 0.980±0.010, 0.640±0.030, and 0.980±0.010, respectively. The above indicators of the model were 0.670±0.180, 0.530±0.200, 0.980±0.010, 0.600±0.040, and 0.990±0.010 when performing anatomical recognition on images with only the left hepatic vein, and 0.580±0.180, 0.430±0.190, 0.980±0.010, 0.580±0.020, and 0.990±0.010 when per-forming anatomical recognition on images with both the left lateral lobe liver pedicle and left hepatic vein.Conclusion:The major anatomical structure recognition model of minimally invasive liver resection based on deep learning can be applied in identifying liver pedicle and hepatic vein.

Pyruvate dehydrogenase E1 component subunit beta-1 (PDHB-1) is a gene encoding the β-subunit of pyruvate dehydrogenase complex, which plays an important role in fruit acid accumulation. The aim of this study was to investigate the evolution characteristics of apple PDHB-1 family and its expression in apples with different acid contents. Bioinformatics analysis was performed using databases including NCBI, Pfam and software including ClustalX, MEGA, and TBtools. By combining titratable acid content determination and quantitative real-time PCR (qRT-PCR), the expression of this family genes in the peel and pulp of apple 'Asda' and 'Chengji No.1' with different acid content were obtained, respectively. The family members were mainly located in chloroplast, cytoplasm and mitochondria. α-helix and random coil were the main factors for the formation of secondary structure in this family. Tissue-specific expression profiles showed that the expression of most members were higher in fruit than in other tissues. qRT-PCR results showed that the expression profile of most members was consistent with the profile of titratable acid contents. In the peel, the expression levels of 14 members in 'Asda' apples with high acid content were significantly higher than that in 'Chengji No.1' apples with low acid content, where the expression difference of MdPDHB1-15 was the most significant. In the pulp, the expression levels of 17 members in 'Asda' apples were significantly higher than that in 'Chengji No.1' apples, where MdPDHB1-01 was the most highly expressed. It was predicted that PDHB-1 gene family in apple plays an important role in the regulation of fruit acidity.
Malus/metabolism* ; Fruit/genetics* ; Protein Structure, Secondary

Objective : To explore the type and control measure of black dot⁃like contaminants in cell cultures. Methods: The growth state of bacteria was investigated under an inverted microscope ; Their morphological characteristics were analyzed by Gram and auramine O staining as well as electron microscopy; 16S rDNA gene sequencing was used to analyze bacterial species ; Drug sensitivity test was used to screen antibiotics against the bacteria;Cryopreserved SH⁃SY5Y cells were resuscitated by cell culture supernatant of RAW264 cells. Results: Inverted microscopic real⁃time observations showed that black dot⁃like substances had two growth states : static and moving.They were negative for Gram staining while positive for auramine O staining. Electron microscopy revealed that they were short rod⁃shaped bacteria with a polar flagellum during moving phase. 16S rDNA gene sequencing showed that these bacteria were phenylobacterium zucineum HLK1. Ceftriaxone , carboxycillin and imipenem were screened by drug sensitivity test to have inhibitory effects on the bacteria , but cell culture experiments showed that they could not remove the bacteria from SH⁃SY5Y cells. Contaminated cells could not be cryopreserved for a long time , but resuscitation with RAW264. 7 cell culture supernatant significantly improved the survival rate of cells. Conclusion The black dot⁃like contaminants in cell cultures are a special type of oligotrophic bacterium with strong viability that can invade the cells and cannot be cleared with antibiotic treatment. RAW264. 7 cell culture supernatant seems contain some substances against bacteria , and resuscitating frozen cells with RAW264. 7 cell culture supernatant may significantly improve the survival rate of cells.

Objective:To investigate the application value of augmented reality navigation combined with indocyanine green(ICG) fluorescence imaging technology in laparoscopic anatomical segment 8 liver resection.Methods:Clinical and pathological data from 8 patients with hepatocellular carcinoma located in segment 8 of the liver admitted to the First Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University from October 2021 to October 2022 were collected restrospectively. Among them,there were 5 males and 3 females,aged between 40 and 72 years. During the operation,the self-developed laparoscopic augmented reality surgical navigation system was used to integrate the three-dimensional liver model with the laparoscopic scene,and ICG fluorescence imaging technology was used to guide the anatomical liver resection of segment 8. The predicted liver resection volume and actual liver resection volume,related surgical indicators and postoperative complications were analyzed.Results:Among the 8 patients, 4 underwent laparoscopic anatomical segment 8 liver resection,1 underwent laparoscopic anatomical ventral subsegment of segment 8 liver resection,2 underwent laparoscopic anatomical ventral subsegment combined with medial subsegment of segment 8 liver resection, and 1 underwent laparoscopic anatomical dorsal subsegment of segment 8 liver resection. All operations were completed under the guidance of augmented reality navigation combined with ICG fluorescence imaging,without conversion to open surgery. The operation time was (276.3±54.8)minutes(range:200 to 360 minutes). Intraoperative blood loss was (75.0±35.4)ml(range:50 to 150 ml). No blood transfusion was performed during the operation. The length of postoperative hospital stay was (7.6±0.8)days(range:7 to 9 days). There were no deaths or postoperative complications such as bleeding or biliary fistula during the perioperative period.Conclusion:Augmented reality navigation combined with ICG fluorescence imaging technology can guide the implementation of laparoscopic anatomical segment 8 liver resection.

Objective:To investigate the application value of augmented reality navigation combined with indocyanine green(ICG) fluorescence imaging technology in laparoscopic anatomical segment 8 liver resection.Methods:Clinical and pathological data from 8 patients with hepatocellular carcinoma located in segment 8 of the liver admitted to the First Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University from October 2021 to October 2022 were collected restrospectively. Among them,there were 5 males and 3 females,aged between 40 and 72 years. During the operation,the self-developed laparoscopic augmented reality surgical navigation system was used to integrate the three-dimensional liver model with the laparoscopic scene,and ICG fluorescence imaging technology was used to guide the anatomical liver resection of segment 8. The predicted liver resection volume and actual liver resection volume,related surgical indicators and postoperative complications were analyzed.Results:Among the 8 patients, 4 underwent laparoscopic anatomical segment 8 liver resection,1 underwent laparoscopic anatomical ventral subsegment of segment 8 liver resection,2 underwent laparoscopic anatomical ventral subsegment combined with medial subsegment of segment 8 liver resection, and 1 underwent laparoscopic anatomical dorsal subsegment of segment 8 liver resection. All operations were completed under the guidance of augmented reality navigation combined with ICG fluorescence imaging,without conversion to open surgery. The operation time was (276.3±54.8)minutes(range:200 to 360 minutes). Intraoperative blood loss was (75.0±35.4)ml(range:50 to 150 ml). No blood transfusion was performed during the operation. The length of postoperative hospital stay was (7.6±0.8)days(range:7 to 9 days). There were no deaths or postoperative complications such as bleeding or biliary fistula during the perioperative period.Conclusion:Augmented reality navigation combined with ICG fluorescence imaging technology can guide the implementation of laparoscopic anatomical segment 8 liver resection.

ObjectiveTo investigate the late detection of HIV/AIDS cases in the elderly in Jilin Province and analyze its influencing factors， to provide theoretical basis for improving their life quality. MethodsThe first CD4 values of HIV/AIDS patients aged 50 years and above living in Jilin Province were used to estimate late detection， and the influencing factors of late detection in elderly cases were analyzed. ResultsThe average CD4 cell count of newly reported HIV/AIDS cases aged 50 and above in Jilin Province from 1996 to 2021 was （230.55±191.97）， the low value group accounted for the largest proportion （50.8%）， and the late detection rate was 59.3% （1397/2325）. The late detection cases were mainly from sexual transmission （46.8% for same-sex and 48.2% for heterosexual contact）. From the perspective of sample sources， most of the late detection patients were diagnosed while testing for other illnesses， followed by testing and consulting. In terms of contact history， the late detection of cases of men who have sex with men was higher. The binary logistic regression analysis showed that gender， marriage， sample source and report year were the factors affecting the late detection of AIDS. The late detection rate of males was higher， and cases of married couples were more likely to be late detection. With the increase of report year， the late detection rate decreased， and testing and counseling could effectively reduce the late detection rate of AIDS. ConclusionThe CD4 cell count in the first detection of HIV/AIDS in the elderly in Jilin Province is low， and the late detection rate of male cases is high. In recent years， the expansion of voluntary counseling and testing in Jilin Province has effectively reduced the late detection rate of HIV/AIDS. At the same time， sex education should be strengthened for the elderly， healthy marital relationships should be advocated and more attention should be paid to the mental health of the elderly.

Objective:To analyze clinical data of 3 children with LMX1B-associated disease characterized by asymptomatic glomerular proteinuria, thus improving the recognition of asymptomatic proteinuria with genetic causes. Methods:Three patients with LMX1B-associated disease presented with prominent asymptomatic proteinuria diagnosed by the next-generation sequencing in Department of Pediatrics, Peking University First Hospital from April 2014 to October 2017 were included in this study.Clinical data, including renal and extrarenal manifestations, renal biopsy, and family history, were collected and retrospectively analyzed. Results:All 3 children were girls, the age of onset were 2 years, 1 year, and 4 years, respectively, and the diagnosis age were 11 years, 5 years and 6 years, respectively.All of them had glomerular proteinuria, and nephrotic-level proteinuria occurred in one patient.Microscopic hematuria was found in 2 patients.All of them had normal renal function.Only one patient underwent renal biopsy.Electron microscopy of the first time of biopsy revealed segmental thinning of the glomerular basement membrane.Re-biopsy 4 years later showed irregular thickening of the glomerular basement membrane, moth-eaten appearance and collagen fibrillar material deposition.No abnormalities of nails, limbs and joints were observed by physical examination.Two patients had a family history of renal disease.Conclusions:Genetic factors should be considered in children with obscure onset asymptomatic proteinuria without definite clinical causes.Genetic testing can help diagnose and guide treatment as early as possible.

Objective    To reveal and demonstrate the hotspots and further research directions in screening technology for early lung cancer, and provide references for the future studies. Methods    Researches related to lung cancer screening from 2011 to 2021 in the Web of Science database were included. Biblioshiny, a bibliometrics program based on R language, was used to perform content analysis and visualization of the included literature information. Results    Researches related to lung cancer screening were increasing year by year. Six major cooperation groups were formed between countries. The current research hotspots in the field of early lung cancer screening technology mainly focused on the multi-directional fusion of radiographic imaging, liquid biopsy and artificial intelligence. Conclusion    Low-dose spiral CT screening is still the most important and mainstream method for the screening of early lung cancer at present. The combination and integration of artificial intelligence with various screening methods and the innovation of novel testing and diagnostic equipment are the current research hotspots and the future research trend in this field.

To Track the source of the infection through an investigation of a clustering of coronavirus disease 2019(COVID-19), and provide scientific basis and Strategy for the effective control of the aggregated epidemic situation of COVID-19. Field epidemiological method was used to survey the cases and related close contacts in a family clustering epidemic of COVID-19 in Dandong city of Liaoning Province. We obtained survey data for a descriptive analysis.Real time RT-PCR technique was used to detect 2019-nCoV nucleic acid in samples collected from cases and related close contacts combined with serum specific antibody detection. A total of 3 confirmed cases and 2 asymptomatic infection cases were discovered in the clustering epidemic, with 34 close contacts.Of eight close family contacts visiting from other province, one patient was on the same flight as the confirmed case, and her antibody IgG was positive. The family clustering was caused by past infection case who visited her friend through Wuhan from other provinces to local area.