Objective: To investigate the objective characteristics of tongue manifestation in different stages of damp-heat syndrome in diabetic kidney disease (DKD). Methods: A cross-sectional study enrolled 134 patients with DKD G3-5 stages who met the diagnostic criteria for damp-heat syndrome in DKD. The patients were treated at Dongzhimen Hospital, Beijing University of Chinese Medicine, from May 2023 to January 2024. The patients were divided into three groups: DKD G3, DKD G4, and DKD G5 stage, with 53, 33, and 48 patients in each group, respectively. Clinical general data (gender, age, and body mass index) and damp-heat syndrome scores were collected from the patients. The YZAI-02 traditional Chinese medicine (TCM) AI Tongue Image Acquisition Device was used to capture tongue images from these patients. The accompanying AI Open Platform for TCM Tongue Diagnosis of the device was used to analyze and extract tongue manifestation features, including objective data on tongue color, tongue quality, coating color, and coating texture. Clinical data and objective tongue manifestation characteristics were compared among patients with DKD G3-5 based on their DKD damp-heat syndrome status. Results: No statistically significant difference in gender or body mass index was observed among the three patient groups. The DKD G3 stage group had the highest age (P<0.05). The DKD G3 stage group had a lower score for symptoms of poor appetite and anorexia(P<0.05) than the DKD G5 group. No statistically significant difference was observed in damp-heat syndrome scores among the three groups. Compared with the DKD G5 stage group, the DKD G3 stage group showed a decreased proportion of pale color at the tip and edges of the tongue (P<0.05). The DKD G4 stage group exhibited an increased proportion of crimson at the root of the tongue, a decreased proportion of thick white tongue coating at the root, a decreased proportion of pale color at the tip and edges of the tongue, an increased hue value (indicating color tone) of the tongue color in the middle, an increased brightness value (indicating color lightness) of the tongue coating color in the middle, and an increased thickness of the tongue coating (P<0.05). No statistically significant difference was observed in other tongue color proportions, color chroma values, body characteristics, coating color proportions, coating color chroma values, and coating texture characteristics among the three groups. Conclusion Tongue features differ in different stages of DKD damp-heat syndrome in multiple dimensions, enabling the inference that during the DKD G5 stage, the degree of qi and blood deficiency in the kidneys, heart, lungs, liver, gallbladder, spleen, and stomach is prominent. Dampness is more likely to accumulate in the lower jiao, particularly in the kidneys, whereas heat evil in the spleen and stomach is the most severe. These insights provide novel ideas for the clinical treatment of DKD.

Objective　To construct a recombinant lentiviral expression vector for human lymphocyte activation gene 3 (LAG3) and generation of monoclonal cell lines that preferentially express LAG3 by transfection of the vector into mouse fibroblast cells 3T3. Methods　After extracting total RNA extracted from human peripheral blood mononuclear cells, the RNA is reversely transcribed into cDNA. The LAG3 extracellular and transmembrane region sequences are amplified by PCR using high-fidelity DNA polymerase. The PCR products are double-digested with the restriction endonucleases EcoRⅠ and NotⅠ, then ligated with the lentiviral vector pTSB-copGFP to construct the recombinant expression vector pTSB-LAG3-copGFP, which is subsequently transformed into Escherichia coli DH5α. Positive clonal bacteria are selected by PCR, and the plasmids are extracted and sequenced for verification. The recombinant vector pTSB-LAG3-copGFP, along with packaging plasmids psPAX2 and pMD2.0G, are co-transfected into human embryonic kidney 293T cells to assemble and release virus particles, the virus infected 3T3 cells were collected. During the puromycin selection of infected 3T3 cells, the limited dilution method is used to obtain 3T3 monoclonal cells that stably express LAG3. Real-time fluorescent quantitative PCR, immunofluorescence and flow cytometry were utilized to verify the transcription of LAG3 mRNA and the expression of LAG3 protein respectively. Results　Sequencing of the recombinant pTSB-LAG3-copGFP lentiviral vector plasmid reveals that the amplified LAG3 sequence contains a synonymous mutation in the His codon at nucleotide position 1 697 bp within the LAG3 transmembrane region, which aligns with the standard LAG3 sequence (accession number NM_002286.6) in GenBank. The 3T3 cells infected by pTSB-LAG3-copGFP packaging virus screened with puromycin. A total of 20 LAG3+copGFP+-3T3 monoclonal cell lines were obtained, all of which exhibited transcription of LAG3 mRNA. The monoclonal cell line MC-6 exhibits the highest transcriptional level of LAG3. Effective expression and distribution of LAG3 protein on the cell membrane and cytoplasmic organelle membranes in MC-6 indicated by immunofluorescence and flow cytometry. Conclusion　The pTSB-LAG3-copGFP lentiviral vector was successfully constructed. LAG3+copGFP+-3T3 monoclonal cell lines overexpressing lymphocyte activating 3 were efficiently established, laying the foundation for subsequent studies on the relationship between LAG3 and the development of chronic infectious diseases such as hepatitis B, as well as the interventional treatment of LAG3.

Periodontal bone defects, primarily caused by periodontitis, are highly prevalent in clinical settings and manifest as bone fenestration, dehiscence, or attachment loss, presenting a significant challenge to oral health. In regenerative medicine, harnessing developmental principles for tissue repair offers promising therapeutic potential. Of particular interest is the condensation of progenitor cells, an essential event in organogenesis that has inspired clinically effective cell aggregation approaches in dental regeneration. However, the precise cellular coordination mechanisms during condensation and regeneration remain elusive. Here, taking the tooth as a model organ, we employed single-cell RNA sequencing to dissect the cellular composition and heterogeneity of human dental follicle and dental papilla, revealing a distinct Platelet-derived growth factor receptor alpha (PDGFRA) mesenchymal stem/stromal cell (MSC) population with remarkable odontogenic potential. Interestingly, a reciprocal paracrine interaction between PDGFRA⁺ dental follicle stem cells (DFSCs) and CD31⁺ Endomucin⁺ endothelial cells (ECs) was mediated by Vascular endothelial growth factor A (VEGFA) and Platelet-derived growth factor subunit BB (PDGFBB). This crosstalk not only maintains the functionality of PDGFRA⁺ DFSCs but also drives specialized angiogenesis. In vivo periodontal bone regeneration experiments further reveal that communication between PDGFRA⁺ DFSC aggregates and recipient ECs is essential for effective angiogenic-osteogenic coupling and rapid tissue repair. Collectively, our results unravel the importance of MSC-EC crosstalk mediated by the VEGFA and PDGFBB-PDGFRA reciprocal signaling in orchestrating angiogenesis and osteogenesis. These findings not only establish a framework for deciphering and promoting periodontal bone regeneration in potential clinical applications but also offer insights for future therapeutic strategies in dental or broader regenerative medicine.
Receptor, Platelet-Derived Growth Factor alpha/metabolism* ; Humans ; Neovascularization, Physiologic/physiology* ; Dental Sac/cytology* ; Single-Cell Analysis ; Transcriptome ; Mesenchymal Stem Cells/metabolism* ; Bone Regeneration ; Animals ; Dental Papilla/cytology* ; Periodontium/physiology* ; Stem Cells/metabolism* ; Regeneration ; Angiogenesis

Patients with chronic obstructive pulmonary disease have the disease phenomenon of fear of exercise because of dyspnea, which can accelerate the body degradation rate, weaken muscle strength, reverse increase dyspnea, and delay the recovery of the disease. As a result, this article examines the theoretical underpinnings and specific measures of dyspnea belief intervention programs for chronic obstructive pulmonary disease patients at home and abroad, summarizes the limitations of previous studies, and makes pertinent recommendations in an effort to serve as a guide for early patient prevention and the development of scientific and feasible intervention programs.

Objective To investigate the clinical value of extracellular volume(ECV)based on CT delayed phase in combination with pathologic indicators in predicting early recurrence of gastric mesenchymal tumors after surgery.Methods A retrospective case-control trial was conducted on the imaging,clinical and pathological data of 110 patients with gastric mesenchymal tumors who were surgically resected at the First Affiliated Hospital of Army Medical University from January 2011 to August 2022.They were 60 males and 50 females,at a mean age of 58±10 years.All of them received preoperative multiphase dynamic CT enhancement examination of the abdomen,and ECV value was calculated with the formula:ECV=(1-hematocrit)×(△HU tumor/△HU aorta).According to the postoperative recurrence within 24 months after surgery,they were divided into early recurrence group and non early recurrence group.Statistical indexes:① Consistency analysis.② The factors affecting early recurrence after resection of gastric mesenchymal stromal tumors were analyzed and a prediction model was conducted.Delong test was used to assess the predictive value of the model.Then a nomogram was plotted based on the combines model,and calibration curves were drawn to assess the efficacy of the column charts,and decision curve analysis(DCA)was adopted to assess the value of the model for clinical application.Results ① Consistency analysis.After 2 radiologists outlined the region of interest and obtained ECV value according to the above formula,The intraclass correlation coefficient(ICC)was 0.806.② For the 110 subjected patients,21 cases of them had early recurrence,and 89 one did not.Multivariate analysis showed that ECV value,risk degree,and tumor length were independent influencing factors for predicting early recurrence.Receiver operating characteristic(ROC)curve analysis indicated that the area under the curve(AUC)value of ECV,hazard degree,and tumor length diameter in predicting early recurrence was 0.838(95％CI 0.758～0.918),0.774(95％CI 0.656～0.892),and 0.700(95％CI 0.589～0.810),respectively,and the value of their combined model was 0.899(95％CI 0.811～0.987),which was higher than that of each independent model.The sensitivity and specificity of the combined model was 85.71％and 86.52％,respectively,and the optimal cutoff value was 0.19.Delong test revealed that there was statistical difference between the combined model and the clinical model established by the hazard level(Z=6.548,P＜0.001,95％CI 0.140～0.259).Calibration curve analysis suggested that the combined model had a better fit,and DCA displayed that the combined model had a better net benefit.Conclusion The model established by ECV combined with pathological indicators has good predictive performance and can be used as a more effective predictor of early recurrence of gastric mesenchymal tumors after surgery.

Objective To prepare mouse anti-human lymphocyte activation gene 3 (LAG3) monoclonal antibody (mAb) and perform immunological identification of the antibody. Methods BALB/c mice were immunized with LAG3-mLumin-3T3 cells, which stably express the extracellular and transmembrane regions of human LAG3 in mouse 3T3 cells. The secretion of anti-human LAG3 antibodies in mouse serum was assessed using flow cytometry and immunofluorescence. SP2/0 cells were injected subcutaneously into the mice to elicit solid myelomas, and mouse myeloma cells were subsequently isolated. Spleen cells from the immunized mice were fused with the myeloma cells to establish hybridomas, which were then separated using the limiting dilution method. Flow cytometry was used to detect LAG3 mAbs in the hybridoma culture medium. To map the epitopes recognized by these mAbs, 3T3 cells expressing individual extracellular domains of LAG3(LAG3 domains 1/-2/-3/-4-3T3) were used. Flow cytometry was also applied to analyze LAG3 expression on activated human peripheral blood mononuclear cells (PBMC) before and after co-culture with the LAG3 mAbs. Results Mice immunized with the recombinant eukaryotic cell antigen produced anti-LAG3 antibodies. The generated hybridomas secreted mouse anti-human LAG3 mAbs, with each hybridoma line recognizing different LAG3 antigenic domains. Conclusion Mouse anti-human LAG3 mAbs were successfully generated, with different hybridoma clones secreting antibodies that recognize distinct LAG3 epitopes. These findings lay the groundwork for further studies into the biological properties of LAG3 and the development of diagnostic reagents and therapeutic blocking antibodies for cancer treatment.
Animals ; Humans ; Mice ; Lymphocyte Activation Gene 3 Protein ; Antibodies, Monoclonal/immunology* ; Mice, Inbred BALB C ; Hybridomas/immunology* ; Antigens, CD/genetics* ; Immunization ; Recombinant Proteins/immunology* ; Female ; Eukaryotic Cells/immunology* ; Flow Cytometry ; Epitopes/immunology*

Objective: To describe the current status of the prevalence of co-morbid myopia and obesity among 7-18 years students in the Inner Mongolia Autonomous Region in 2021 and to analyze the moderating effect of lifestyle in this association ，so as to provide scientific basis for the establishment of the mechanism of Co-morbidity，Shared Etiology，and Shared Prevention of common diseases in children and adolescents. Methods: A total of 139 630 primary and secondary school students aged 7-18 years from Inner Mongolia Autonomous Region were selected by stratified random cluster sampling method in September,2021. Myopia was determined using distance visual acuity examination and refractive error examination, and obesity was determined according to the BMI classification criteria for overweight, obesity screening of Chinese school age children and adolescents. Used a questionnaire, healthy lifestyles were determined according to the American Heart Association s Healthy Lifestyle Score by totaling the six scores for smoking, alcohol consumption, diet, exercise, screen time, and sleep duration.The χ 2 test was used to compare the association between group differences in the co-morbidity rate of myopia and obesity. The multivariable Logistic regression model was used to explore the influencing factors of the co-morbidity of myopia and obesity, and the stratified analysis was used to analyze the moderating effect of lifestyles on the prevalence of the co-morbidity. Results: The prevalence of myopia and obesity co-morbidity among students aged 7-18 years old in the Inner Mongolia Autonmous Region in 2021 was 13.7%, higher among boys than girls ( 15.5 % vs. 11.8%), higher among those aged 10-12 years old than 7-9,13-15,and 16-18 years old (14.7%,13.7%, 13.3%, 12.0%), higher among other ethnic minorities than Han Chinese and Mongolians (15.3%, 14.0%, 12.5%), higher in urban areas than that in suburban areas(15.3%, 13.0%), and middle economic level tracts were higher than poor and good tracts (14.8%, 12.9 %, 12.6%) ( χ 2=392.37，115.73，62.80，119.02，121.60, P <0.05). Multivariable Logistic regression modeling showed that unhealthy lifestyles ( OR=1.24, 95%CI=1.19-1.29 ) and middle level of lifestyle score ( OR=1.15, 95%CI=1.10-1.19 ) students had higher prevalence of co-morbidity, and the results were statistically significant among both boys and girls, the age groups of 10- 12, 13-15, and 16-18 years old, as well as the Han and Mongolian ethnic groups (all P <0.05). Conclusion In 2021, the current situation of myopia and obesity co-morbidity and unhealthy lifestyles among primary and secondary school students in the Inner Mongolia Autonomous Region are not optimistic.

Objective:To evaluate the safety and efficacy of rubber band and clip facilitated endoscopic submucosal dissection (RAC-ESD) for colorectal neoplasms.Methods:A retrospective cohort study was performed. Clinical data of 115 patients with colorectal neoplasm receiving ESD from September 2018 to August 2019 were retrospectively analyzed. Thirty-four patients received RAC-ESD treatment (RAC-ESD group) and 81 received conventional ESD treatment (conventional ESD group). The procedure time, the dissected area per minute during ESD, en bloc resection rate, complete resection rate, curative resection rate, complication occurence and recurrence rate were compared between the two groups.Results:The median specimen area of RAC-ESD group was 6.32 (7.53) cm 2, and the median procedure time was 40.0 (55.0) min. The mean dissected area per minute was 0.14 (0.20) cm 2/min. While in conventional ESD group, the median specimen area was 4.71 (5.02) cm 2, the median procedure time was 50.0 (50.0) min and the mean dissected area per minute was 0.09 (0.07) cm 2/min. The median specimen area of RAC-ESD group was slightly larger and the median procedure time was slightly shorter than those of conventional ESD group, but neither was significantly different(both P>0.05). The median dissected area per minute of RAC-ESD group was significantly larger than that of the conventional ESD group ( P=0.008). The en bloc resection rate, complete resection rate and curative resection rate of RAC-ESD group were 100.0% (34/34), 100.0% (34/34) and 97.1% (33/34), while those of the conventional ESD group were 100.0%(81/81), 96.3%(78/81) and 91.4%(74/81), respectively. There was no ESD-related complication in either group. After 10.0±5.5 months of follow-up, there was no local recurrence in both groups. Conclusion:RAC-ESD may increase resection efficacy with safety.

Objective? To investigate the effect and safety of high-flow nasal cannula (HFNC) oxygen therapy for patients after extubation in lung transplantation. Methods? A retrospective study was conducted on 60 hospitalized patients with lung transplant during January 2017 to December 2017 in Wuxi People＇s Hospital. According to the different methods of respiratory support prescribed after extubation, the patients were divided into two groups: observation group (n=28, via HFNC) and control group (n=32, via nasal oxygen tube) . The two groups were compared in terms of the clinical indicators including their blood gas analysis(oxygenation index, lactic acid, partial pressure of carbon dioxide) , viscosity of sputum and comfort, etc. Results? Six hours after extubation and before transferring to other departments, the observation group＇s oxygenation index was(263.70±48.97)and(273.22±43.26)mmHg, which were statistically different from those of the control group with (217.83±77.30)and(229.08±68.64)mmHg respectively (P＜ 0.05). Before transferring to other departments, the partial pressure of carbon dioxide in the observation group was (37.04±8.56)mmHg, lower than the control group with (42.43±6.14)mmHg with a statistical difference (P＜0.05); the viscosity of sputum in the obseration group was lower than the control group with statistical difference (P＜0.05); the patients＇ comfort in the observation group achieved (7.72±1.06)points, higher than the control group＇s (4.39±0.82)points with statistical difference (P＜ 0.05). Conclusions? HFNC oxygen therapy for the patients after extubation in lung transplantation has good effects, is safe and reliable, and can be widely applied in clinical practice.

This study was aimed to investigate the effects of Si-Wu mixture on the secretion of E2 and the expression of CYP19al gene in granulosa cells after cell injury.Ovarian granulosa cells of SD rats were treated with cisplatin (CDDP).And the expression levels of E2 and CYP19a1 were determined by different testing methods.The results showed that the level of E2 induced by radioimmunoassay of the Si-Wu group was significantly higher than that of the CDDP group.Meanwhile,the group which added TGF-β3 protein pathway blocker was lower than others.The results of immunohistochemistry and western blotting showed that the expression level of CYP 19a1 of the Si-Wu group was higher than that of the CDDP group.In the western blotting,the group which added blocker was significantly lower than the non-blocking group.It was concluded that the pharmacological serum of Si-Wu mixture can enhance the level of E2 in CDDP cells through TGF-β3 protein pathway.And the effect is accomplished by the intervention of CYP 19a1.