Objective:To investigate the effect of Hong's One Stitch Method in pancreaticoduodenectomy(PD).Methods:A total of 40 patients who underwent PD in our hospital from Jan 2021 to Dec 2022 were divided into two groups according to random number table method,with 20 patients in each group.The control group was treated with end to end pancreatojejunal anastomosis,and the observation group was treated with"Hong's One Stitch Method".The perioperative indicators,complications,secondary surgery,mortality and quality of life were compared between the two groups.Results:The pancreatoenteroanastomosis time,operation time and hospitalization time in the observation group were shorter than those in the control group,and the incidence of pancreatic fistula was lower than that in the control group(P＜0.05).There were no significant differences in intraoperative blood loss,pancreatic biochemical leakage,bile fistula,hemorrhage,localized abdominal infection,gastric emptying obstruction,pulmonary infection,secondary surgery and mortality between the two groups(P＞0.05).The mental health score,emotional function score,social function score,energy score,general health status score,body pain score,and physiological function score in the observation group were higher than those in the control group(P＜0.05).Conclusion:In PD surgery,the application of"Hong's One Stitch Method"to perform pancreatoenterostomy is beneficial to shorten the pancreatoenterostomy time,operation time and hospitalization time,accelerate the postoperative recovery,reduce the incidence of pancreatic fistula,and improve the quality of life of patients.

Objective:To construct a microfluidic organ-on-a-chip and evaluate its capability in simulating subchondral bone remodeling during the progression of osteoarthritis.Methods:The chip′s main body was designed based on the microfluidic technology and cell co-culture technique. MC3T3-E1 cells were cultured adherently within the cell seeding micro-chamber, with the culture medium perfused at a flow rate of 0.5 ml/min at the bottom of the micro-chamber. Evaluation metrics were as follows: (1) Assessment of the microfluidic organ-on-a-chip: The growth culture medium was perfused and simulation experiments were conducted to test the concentration differences and equilibrium times of the fluid inside and at the bottom of the cell seeding micro-chamber at various time points; live-dead staining was performed to observe the biocompatibility of cells cultured continuously for 3 days and 7 days at a set flow rate, which was divided into 3-day and 7-day groups. (2) Osteogenic potential of the microfluidic organ-on-a-chip: The osteogenic induction medium was perfused, and ALP staining and PCR were performed to compare the number of the black alkaline phosphatase (ALP)-positive cells and the expression levels of osteogenesis-related marker genes including osteoblast-specific transcription factor 2 (RUNX2), type I collagen (COL1A1), bone morphogenetic protein-2 (BMP-2), and osteocalcin (OCN) under static, 3-day and 7-day perfusion conditions, which was divided into static non-induced, static-induced and perfusion-induced groups. (3) Characterization of morphology and size, and biocompatibility of extracellular vesicles (EVs) of three osteoblast subtypes: Three different subtypes of osteoblasts were obtained [endothelial-type osteoblasts (EnOB)-EVs, stromal-type osteoblasts (StOB)-EVs and mineralizing-type osteoblasts (MinOB)-EVs]. Their morphology and size were obtained through transmission electron microscopy and particle size analysis. Growth medium containing EVs of three different cell subtypes was perfused, and cell proliferation/apoptosis assay was performed to compare the biocompatibility of the addition of different EVs concentrations (1, 1.25, 2.5, and 5 μg/ml) for 24 hours, which was categorized into the EnOB-EVs group, StOB-EVs group and MinOB-EVs group. (4) Osteogenic effect of EVs from three subtypes of osteoblasts: Osteogenic induction media containing EVs from three different osteoblast subtypes were perfused for 3 days, and ALP staining and PCR were performed to compare the number of black ALP-positive cells and the expression levels of osteogenesis-related marker genes including RUNX2, COL1A1, BMP-2, and OCN, which was divided into non-EVs group, EnOB-EVs group, StOB-EVs group and MinOB-EVs group.Results:(1) Evaluation of the microfluidic organ-on-a-chip: Simulation results showed that the concentration in the top layer of the upper chamber reached more than 95% of that in the lower chamber and that the concentration in the bottom layer was about 96.5% of that in the lower chamber after 12 hours of continuous perfusion, reaching an equilibrium state of the concentration difference between the upper and lower chambers. The results of live-dead staining showed that the chip was biocompatible at a flow rate of 0.5 ml/min, and the cell survival rate at 3 and 7 days of perfusion was (99.48±0.12)% and (97.07±1.05)% ( P<0.01). (2) ALP staining results showed that at 3 days, the perfusion-induced group showed the highest number of black ALP-positive cells, followed by the static-induced group, and the least in the static non-induced group. At 7 days, the static-induced group had the highest number of black ALP-positive cells, followed by the perfusion-induced group, and the least in the static non-induced group. PCR results indicated that at 3 days, the expression levels of RUNX2, COL1A1, BMP-2, and OCN were 1.00±0.03, 1.00±0.12, 1.00±0.01, and 1.00±0.02 respectively in the static non-induced group; 1.80±0.04, 4.05±0.37, 9.80±1.94, and 4.38±0.89 respectively in the static-induced group, and 2.45±0.23, 5.48±0.42, 91.50±4.56, and 10.82±4.96 respectively in the perfusion-induced group ( P<0.01). At 7 days, the expression levels of RUNX2 was 1.00±0.01 in the static non-induced group, 1.46±0.46 in the static-induced group, and 1.11±0.08 in the perfusion-induced group ( P>0.05); the expression levels of COL1A1, BMP-2, and OCN were 1.00±0.03, 1.00±0.13, and 1.00±0.09 respectively in the static non-induced group, 9.38±0.25, 14.27±4.35, and 84.01±4.02 respectviely in the static-induced group, and 2.39±0.08, 133.64±8.87, and 86.64±8.36 respectively in the perfusion-induced group ( P<0.01). When comparing the static non-induced, static-induced, and perfusion-induced groups at both 3 and 7 days, the perfusion-induced group demonstrated the strongest osteogenic capability. (3) Characterization of morphology and size and biocompatibility of EVs from three osteoblast subtypes: Under the transmission electron microscope, EVs from EnOB-EVs, StOB-EVs, and MinOB-EVs all exhibited a typical saucer-shaped morphology. The particle sizes of EnOB-EVs, StOB-EVs, and MinOB-EVs were (91.3±14.7)nm, (106.0±16.0)nm, and (68.1±10.7)nm, respectively. Cell proliferation/apoptosis assay results indicated that the optimal administration concentration of EnOB-EVs, StOB-EVs, and MinOB-EVs was all 1.25 μg/mL. (4) Validation of osteogenic effect of the microfluidic organ-on-a-chip on three types of EVs: ALP staining results showed that the non-EVs group had the fewest black ALP-positive cells, followed by the EnOB-EVs group, then the StOB-EVs group, and the MinOB-EVs group had the most. PCR results showed that the expression levels of RUNX2, COL1A1, BMP-2, and OCN were 1.00±0.01, 1.00±0.03, 1.00±0.02, and 1.00±0.02 respectively in the non-EVs group, 1.95±0.11, 6.78±2.04, 7.99±0.57, and 6.93±3.83 repectively in the EnOB-EVs group, 0.79±0.12, 5.68±1.53, 12.59±3.15, and 25.59±0.95 respectively in the StOB-EVs group, and 0.68±0.10, 4.36±0.69, 18.75±3.21, and 34.74±3.98 repectively in the MinOB-EVs group ( P<0.01). Compared with the non-EVs group, EnOB-EVs group, StOB-EVs group, and MinOB-EVs group, the MinOB-EVs group showed the most significant osteogenic effect. Conclusions:The microfluidic organ-on-a-chip constructed using microfluidic technology and cell co-culture techniques is capable of maintaining the normal growth of MC3T3-E1 cells, enhancing their proliferation and osteogenic induction differentiation. EVs released by osteoblasts at different stages possess osteogenic effects and can accelerate the bone sclerosis in the remodeling of subchondral bone during the progression of osteoarthritis.

Objective To explore the correlation between polycystic ovary syndrome(PCOS)and periodontitis in light of cytokines levels,sex hormone levels and metabolism-related indicators and their changes during progression of the two diseases.Methods Twenty healthy subjects and 40 patients diagnosed with PCOS underwent full-mouth periodontal examinations to obtain full-mouth plaque score(FMPS),gingival bleeding index of probing(BOP),probing depth(PD),and clinical attachment level(CAL).The participants were divided into Group A without periodontitis or PCOS(n=15),Group B with PCOS but without periodontitis(n=28),Group C with periodontitis but without PCOS(n=5),and Group D with both diseases(n=12).Serum levels of luteinizing hormone/follicle stimulating hormone(LH/FSH),testosterone,prolactin,progesterone and estradiol,and the levels of interleukin 6(IL-6),IL-17A,tumor necrosis factor α and matrix metalloproteinase 8(MMP-8)in both serum and saliva samples were measured at the time of enrolment and at 3 and 6 months after enrolment and compared among the 4 groups.Results Serum MMP-8 level was significantly higher in Group B than in Group A(P<0.05).Salivary MMP-8 level was significantly higher in Group D than in Group B(P<0.05).Salivary MMP-8,LH,and LH/FSH levels and serum and salivary IL-6 and progesterone levels all tended to increase in the 6 months after enrollment(OR>1,P<0.05).During the follow-up period,serum IL-6 levels differed significantly between the non-PCOS groups(A and C)and PCOS groups(B and D)(P<0.05);serum IL-6 and salivary MMP-8 levels differed significantly between the non-periodontitis groups(A and B)and periodontitis groups(C and D)(P<0.05).Spearman correlation analysis indicated positive correlations of LH and LH/FSH with PD(P<0.05);testosterone and LH/FSH were positively correlated with serum MMP-8 levels(P<0.05),and PD,BOP and FMPS were positively correlated with salivary MMP-8 levels(P<0.01).Conclusion There is a correlation between PCOS and periodontitis,and their progression is accompanied by changes in serum and salivary levels of pro-inflammatory cytokines and serum sex hormones.

Objective To explore the correlation between polycystic ovary syndrome(PCOS)and periodontitis in light of cytokines levels,sex hormone levels and metabolism-related indicators and their changes during progression of the two diseases.Methods Twenty healthy subjects and 40 patients diagnosed with PCOS underwent full-mouth periodontal examinations to obtain full-mouth plaque score(FMPS),gingival bleeding index of probing(BOP),probing depth(PD),and clinical attachment level(CAL).The participants were divided into Group A without periodontitis or PCOS(n=15),Group B with PCOS but without periodontitis(n=28),Group C with periodontitis but without PCOS(n=5),and Group D with both diseases(n=12).Serum levels of luteinizing hormone/follicle stimulating hormone(LH/FSH),testosterone,prolactin,progesterone and estradiol,and the levels of interleukin 6(IL-6),IL-17A,tumor necrosis factor α and matrix metalloproteinase 8(MMP-8)in both serum and saliva samples were measured at the time of enrolment and at 3 and 6 months after enrolment and compared among the 4 groups.Results Serum MMP-8 level was significantly higher in Group B than in Group A(P<0.05).Salivary MMP-8 level was significantly higher in Group D than in Group B(P<0.05).Salivary MMP-8,LH,and LH/FSH levels and serum and salivary IL-6 and progesterone levels all tended to increase in the 6 months after enrollment(OR>1,P<0.05).During the follow-up period,serum IL-6 levels differed significantly between the non-PCOS groups(A and C)and PCOS groups(B and D)(P<0.05);serum IL-6 and salivary MMP-8 levels differed significantly between the non-periodontitis groups(A and B)and periodontitis groups(C and D)(P<0.05).Spearman correlation analysis indicated positive correlations of LH and LH/FSH with PD(P<0.05);testosterone and LH/FSH were positively correlated with serum MMP-8 levels(P<0.05),and PD,BOP and FMPS were positively correlated with salivary MMP-8 levels(P<0.01).Conclusion There is a correlation between PCOS and periodontitis,and their progression is accompanied by changes in serum and salivary levels of pro-inflammatory cytokines and serum sex hormones.

The accurate deduction of postmortem interval is a difficult and extremely critical task in the field of forensic pathology and criminal investigation.In recent years,physics,chemistry,microbiology,immunology,entomology,molecular biology and other disciplines have been on the rise,and special staining,spectroscopy,mass spectrometry,chromatography,radiographic and other techniques have been developed,and methods for postmortem interval inference are increasing,among which immunolabelling techniques have played an important role.In this paper,we systematically reviewed the domestic and foreign relevant studies on the postmortem interval inference using of immunolabelling techniques,including immunohistochemistry,immunoblotting,immunofluorescence,radioimmunoassay,etc.We summarized and analyzed the research progress on these techniques in postmortem interval inference,with the aim of exploring the ideas for the study of postmortem interval inference in forensic pathology,and provide reference for better applying them in forensic practice.

Objective:To investigated the safety and efficacy of donor-derived CD19+ or sequential CD19+ CD22+ chimeric antigen receptor T-cell (CAR-T) therapy in patients with B-cell acute lymphoblastic leukemia (B-ALL) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The data of 22 patients with B-ALL who relapsed after allo-HSCT and who underwent donor-derived CAR-T therapy at the Zhujiang Hospital of Southern Medical University and the 920th Hospital of Joint Logistics Support Force of the People’s Liberation Army of China from September 2015 to December 2022 were retrospectively analyzed. The primary endpoint was overall survival (OS), and the secondary endpoints were event-free survival (EFS), complete remission (CR) rate, and Grade 3-4 adverse events.Results:A total of 81.82% ( n=18) of the 22 patients achieved minimal residual disease-negative CR after CAR-T infusion. The median follow-up time was 1037 (95% CI 546–1509) days, and the median OS and EFS were 287 (95% CI 132-441) days and 212 (95% CI 120-303) days, respectively. The 6-month OS and EFS rates were 67.90% (95% CI 48.30%-84.50%) and 58.70% (95% CI 37.92%-79.48%), respectively, and the 1-year OS and EFS rates were 41.10% (95% CI 19.15%-63.05%) and 34.30% (95% CI 13.92%-54.68%), respectively. Grade 1-2 cytokine release syndrome occurred in 36.36% ( n=8) of the patients, and grade 3-4 occurred in 13.64% of the patients ( n=3). Grade 2 and 4 graft-versus-host disease occurred in two patients. Conclusion:Donor-derived CAR-T therapy is safe and effective in patients with relapsed B-ALL after allo-HSCT.

Objective To explore the feasibility of quartz glass for radiotherapy dosimetry through the experimental study of the thermoluminescence characteristics of synthetic quartz glass. Methods The thermoluminescence glow curves of quartz glass under different annealing conditions were analyzed, the thermoluminescence characteristics of quartz glass were studied, and the measurement parameters were optimized. Using the Co-60 reference radiation field in the National Secondary Standard Dosimetry Laboratory, the quartz glass samples under different annealing conditions were irradiated following the dose levels of radiotherapy, i.e., 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 10.0 Gy, respectively. According to the relationship between the absorbed dose of quartz glass and the relative thermoluminescence signal intensity, the linearity and dispersion of the dose response of quartz glass were obtained, and the feasibility of quartz glass for radiotherapy dosimetry was analyzed. Results The linear correlation coefficient of dose response of quartz glass under annealing condition of 430℃ for 10 min was 0.9984, and the dose response dispersion was 0.97% at the absorbed dose of 2 Gy. The linear correlation coefficient of dose response of quartz glass under annealing condition of 600℃ for 1 h was 0.9911, and the dose response dispersion was 1.4% at the absorbed dose of 2 Gy. Conclusion Preliminary results suggest that quartz glass with annealing condition of 430℃ for 10 min has the potential to be used for radiotherapy dosimetry.

Objective:This study aims to reveal the special immune infiltrating environment and possible immune escape mechanism of giant cell tumor of bone through single-cell sequencing data.Methods:The fresh samples obtained from 4 patients with primary giant cell tumor of bone from January 2018 to December 2021 were collected, and single-cell transcriptome sequencing was performed on the 10X platform to explore the characteristics and immune environment of giant cell tumor of bone by using t-distributed stochastic neighbor embedding ( t-SNE). The main cell types and signal pathways of immune cell regulation and function in giant cell tumor of bone were observed by cell communication analysis. Results:Cell clustering, the definition of basic cell types, the classification of immune cells, and the mutual regulatory relationship between cell types were analyzed for 35 643 single-cell data from 4 giant cell tumor samples of bone. It was found that giant cell tumor of bone was composed of 9 basic cell types, in which the immune cells were mainly CD8 + T cells (51%) and the non-immune cells were mainly fibroblast like spindle stromal cells (74%). The immune infiltration of giant cell tumor of bone is dominated by cytotoxic CD8 + T cells and lacks exhausted CD8 + T cells. CD4 + T cells are characterized by high expression of immune checkpoint genes CTLA4 and TIGIT. In giant cell tumor of bone, immune cells mainly act on multinucleated osteoclast like giant cells through PARs and CCL signaling pathways, but not stromal cells. Conclusion:This study defined the main cell types of giant cell tumor of bone through single cell sequencing data, and further revealed the composition characteristics of its immune infiltration, and found that the target of its immune cells was mainly multinuclear osteoclast like giant cells, which provided effective information for further understanding the occurrence and development of giant cell tumor of bone.

Long noncoding RNAs (lncRNAs) are a class of RNA molecules that are greater than 200 nt in length and do not have protein-coding capabilities or encode micropeptides only. LncRNAs are involved in the regulation of cell proliferation, differentiation, apoptosis and other biological processes, and are closely associated with the occurrence, recurrence and metastasis of a variety of malignant hematologic diseases. This article summarizes the function, regulatory mechanism and potential clinical application of lncRNAs in leukemia. In general, lncRNAs regulate the occurrence and development of leukemia and the multi-drug resistance in chemotherapy through epigenetic modification, ribosomal RNA transcription, competitive binding with miRNA, modulating glucose metabolic pathway, and activating tumor-related signaling pathway. Studies on lncRNAs provide new references for understanding the pathogenesis of leukemia, uncovering new prognostic markers and potential therapeutic targets, and addressing the problems of drug resistance and post-treatment recurrence in patients in clinical treatment of leukemia.
Cell Proliferation ; Humans ; Leukemia/genetics* ; MicroRNAs ; Neoplasms ; RNA, Long Noncoding/genetics*

Development of thoracolumbar vertebra (TLV) and rib primordium (RP) is a common evolutionary feature across vertebrates, although whole-organism analysis of the expression dynamics of TLV- and RP-related genes has been lacking. Here, we investigated the single-cell transcriptome landscape of thoracic vertebra (TV), lumbar vertebra (LV), and RP cells from a pig embryo at 27 days post-fertilization (dpf) and identified six cell types with distinct gene expression signatures. In-depth dissection of the gene expression dynamics and RNA velocity revealed a coupled process of osteogenesis and angiogenesis during TLV and RP development. Further analysis of cell type-specific and strand-specific expression uncovered the extremely high level of HOXA10 3'-UTR sequence specific to osteoblasts of LV cells, which may function as anti-HOXA10-antisense by counteracting the HOXA10-antisense effect to determine TLV transition. Thus, this work provides a valuable resource for understanding embryonic osteogenesis and angiogenesis underlying vertebrate TLV and RP development at the cell type-specific resolution, which serves as a comprehensive view on the transcriptional profile of animal embryo development.