Objective To investigate the effects of FasL gene-modified dendritic cell (DC) on the airway inflammation in mice sensitized/challenged by house dust mite (HDM) allergen.Methods FasL gene-modified DC (FasL-DC) and control DC (nontransfection DC) were administrated into HDM sensitized and challenged mice by intratracheal injection respectively，then HDM sensitized and challenged mice were sacreificed two days later.Total and differentiation cell counts and levels of interleukin-4（IL-4），IL-5 and interferon-γ(IFN-?) in bronchoalveolar lavage fluid (BALF) were detected and lung histological features were observed.Results After administration of FasL-DC，lung allergic inflammation was ameliorated while total cell counts，the percentage of eosinophil ,the levels of IL-4 and IL-5 in BALF decreased and the level of IFN-? in BALF increased.Conclusion Administrating FasL-DC into HDM sensitized/challenged mice can inhibit Th2 cells activation and ameliorate airway allergic inflammation.
ObjectiveTo compare the short-term effectiveness of suture hook suture via double posteromedial approaches and Fast-Fix total internal suture in treatment of Ramp lesions. Methods A clinical data of 56 patients with anterior cruciate ligament rupture combined with Ramp lesions, who met the selection criteria and admitted between December 2021 and February 2023, was retrospectively analyzed. The Ramp lesions were sutured using suture hook via double posteromedial approaches under arthroscopy in 28 cases (group A) and treated with Fast-Fix total internal suture under arthroscopy in 28 cases (group B). There was no significant difference in age, gender, cause of injury, type of injury, time from injury to operation, side of injury, body mass index, and preoperative Lysholm score, visual analogue scale (VAS) score, and Tegner score between the two groups (P>0.05). The patients were followed up regularly after operation, and the clinical and imaging healing of the Ramp lesion was evaluated according to the Barrett clinical healing standard and the MRI evaluation standard. Lysholm score, VAS score, and Tegner score were used to evaluate the function and pain degree of knee joint, and the results were compared with those before operation. ResultsThe incisions of the two groups healed by first intention. All patients were followed up 12-18 months (mean, 14.9 months). Postoperative McMurray tests were negative in both groups. The clinical healing rates of group A and group B were 71.4% (20/28) and 64.3% (18/28) at 6 months after operation, and 92.9% (26/28) and 82.1% (23/28) at 12 months after operation, respectively. The differences between the two groups was not significant (χ2=0.327, P=0.567; χ2=0.469, P=0.225). There was no significant difference in Lysholm score, VAS score, and Tegner score between the two groups at each time point after operation (P>0.05). The postoperative scores in the two groups significantly improved when compared with those before operation, and the scores at 12 months after operation further improved when compared with those at 6 months after operation, showing significant differences between the different time points in the two groups (P<0.05). At last follow-up, MRI examination of the knee joint showed that there were 26 (92.9%), 2 (7.1%), and 0 (0) cases of complete healing, partial healing, and nonunion in the Ramp lesion of group A, and 25 (89.3%), 1 (3.6%), and 2 (7.1%) cases in group B, respectively. There was no significant difference between the two groups (Z=?0.530, P=0.596). ConclusionSuture hook suture via double posteromedial approaches and Fast-Fix total internal suture under arthroscopy are safe and reliable in the treatment of Ramp lesion, and the knee joint function significantly improves after operation.
ObjectiveTo evaluate the effect of pre-infusion of allogeneic lymphoyctes treated with 5-FU on the rat liver graft. MethodsRat liver transplant models from Wistar to SD were established. Four groups were designed as following: control group: only liver transplantation without any other intervention; lymphocytes group: 1 ml of untreated lymphocytes (5×106/ml) from Wistar rats were preinfused into SD rats on day 7 and 4 separately before transplantation; lymphocytes with low concentration of 5-FU group: low concentration 5-FU (7.5 μg) treated lymphocytes were preinfused as above; lymphocytes with high concentration of 5-FU group: high concentration 5-FU (15 μg) treated lymphocytes were preinfused as above. Fas-L and CD8 expression were detected by immunohistochemistry method on day 7 after transplantation. ResultsThe integral opticaldensity (IOD) of Fas-L positive lymphocytes in the lobules of liver and portal areas were higher in lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. There was no difference between lymphocyte group and lymphocytes with high concentration of 5-FU group （Pgt;0.05）. The IOD of CD8+ expression in lobules of liver was not different among all the three lymphocytes treated groups （Pgt;0.05）. But in portal areas, CD8+ expression was lower in the lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. ConclusionPreinfusion of lymphocytes treated with low concentration 5-FU can induce graft immune tolerance, the probable mecanism of which is the increasing Fas-L expression in graft.
Objective To detect gene mutations of Fas gene death domain （exons 7-9） in 2 Chinese keloid pedigrees and to investigatethe significance of Fas gene mutations in the keloid formation.Methods The samples were selected from keloid pedigrees A and B in 2005. The polymerase chainreaction and DNA sequencing analysis technique were used to detect the sequenceof exons 7-9 of Fas gene from keloid tissues of 2 male patients in pedigree A,their peripheral vein blood and their surrounding normal skin served as their own contrast, their spouses’ peripheral vein blood served as normal contrast, the peripheral vein blood of 2 patients in pedigree B served as a contrast between different keloid pedigrees.Results No gene mutations and single nucleotidepolymorphism in Fas gene exons 7, 8 were found in all samples from pedigrees A and B. But point mutations and single nucleotide polymorphism in Fas gene exon 9were identified in 11 bp and 53 bpin 2 keloid tissue samples from Chinese keloid pedigree A.Conclusion Fas gene point mutations maybe indicate some relations in Fas protein function and keloid formation.
Objective To explore the expressions of Galectin-3, Fascin-1, and β-catenin protein in colorectal adenocarcinoma and the relations to clinicopathologic characteristics. Methods The expressions of Galectin-3, Fascin-1, and β-catenin protein were detected in 60 cases of colorectal adenocarcinoma, 30 cases of adenoma, and 30 cases of normal mucosa by microwave-EliVisionTM immunohistochemistry method, and analyzed the expressions of them and the relations to clinicopathologic characteristics. Results The expression rate of Galectin-3, Fascin-1, and β-catenin protein in CRC was 68.3% （41/60）, 53.3% （32/60）, and 81.7% （49/60） respectively, which was 46.7% （14/30）, 30.0% （9/30）, and 43.3% （13/30） respectively in adenoma, and 20.0% （6/30）, 3.3% （1/30）, and 13.3% （4/30） respectively in normal mucosa, the differences had statistical significance （P＜0.05）. The expressions of Galectin-3, Fascin-1, and β-catenin protein had statistically significant correlation with the TNM stage, invasive degree, and lymph node metastasis of colorectal adenocarcinoma （P＜0.05, P＜0.01）. The expressions of Galectin-3 and β-catenin protein had statistically significant correlation with the different differentiation degree of colorectal adenocarcinoma （P＜0.05）, but the expression of Fascin-1 protein was not related to differentiation degree of colorectal adenocarcinoma （P＞0.05）.The expressions of Galectin-3, Fascin-1, and β-catenin protein had not statistically significant correlation with the patient’s age and gender, and tumour size （P＞0.05）.There were positive correlations between the Galectin-3 and Fascin-1 or β-catenin （r=0.728，P＜0.01；r=0.696，P＜0.01）, and there was positive correlation between β-catenin and Fascin-1 （r=0.507，P＜0.01）. Conclusions The high expressions of Galectin-3, Fascin-1, and β-catenin protein in colorectal adenocarcinoma tissues are some extent correlated to the high invasive ability and lymph node metastasis, which could be used for the indexes to predict the invasion and metastasis in colorectal carcinoma potentially.
【Abstract】ObjectiveTo measure the expressions of Fas/FasL mRNA in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma, and to explore the relationship between the expressions of Fas/FasL mRNA in those tissues and the hepatocellular carcinogenesis. MethodsSemi-quantity reverse transcript-ploymerase chain reaction(QRTPCR) were performed to measure the relative quantity of the Fas and FasL mRNA expressions in normal liver (n=25), adjacent noncancerous liver parenchyma(n=40) and hepatocarcinoma(n=40). ResultsThe relative quantity of Fas and FasL mRNA expressed in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma were 0.792±0.039 vs 0.245±0.043，0.857±0.031 vs 0.429±0.035 and 0.473±0.047 vs 0.185±0.041, respectively. The relative quantity of Fas mRNA expression in hepatocarcinoma was lower than that of normal liver tissue and adjacent non-cancerous liver parenchyrna (P＜0.05). The relative quantity of FasL mRNA expression in hepatocarcinoma was also lower than that of normal liver tissue (P＜0.05) and adjacent non-cancerous liver parenchyma (P＜0.01), but its expression in adjacent non-cancerous liver parenchyma was higher than that of normal liver tissue (P＜0.05).ConclusionHepatorcarcinoma may escape the immune surveillance of the host, not only by means of reducing Fas expression, but also through adjacent non-cancerous liver parenchyma’s increasing expression of FasL to induce apoptosis of contact lymphocyte which highly expresses Fas.
The number of white blood cells in the leucorrhea microscopic image can indicate the severity of vaginal inflammation. At present, the detection of white blood cells in leucorrhea mainly relies on manual microscopy by medical experts, which is time-consuming, expensive and error-prone. In recent years, some studies have proposed to implement intelligent detection of leucorrhea white blood cells based on deep learning technology. However, such methods usually require manual labeling of a large number of samples as training sets, and the labeling cost is high. Therefore, this study proposes the use of deep active learning algorithms to achieve intelligent detection of white blood cells in leucorrhea microscopic images. In the active learning framework, a small number of labeled samples were firstly used as the basic training set, and a faster region convolutional neural network (Faster R-CNN) training detection model was performed. Then the most valuable samples were automatically selected for manual annotation, and the training set and the corresponding detection model were iteratively updated, which made the performance of the model continue to increase. The experimental results show that the deep active learning technology can obtain higher detection accuracy under less manual labeling samples, and the average precision of white blood cell detection could reach 90.6%, which meets the requirements of clinical routine examination.
Objective To replace dysfunctional Fas gene and reconstruct the blocked Fas signal by using two kinds of prepared recombinantAdenovirus which have human Fas gene. Methods After the keloids derived from fibroblasts were infected by the Adenovicus, the expressions of Fas protein before the exposure and after the exposure was compared. Then the function of the newly produced Fas protein was detected. Results The highly improve expression of Fas protein in the infected keloid derived fibroblasts was detected. Obvious apoptosis was also detected in the infected keloid derived from fibroblasts under the condition of exposing to FasMcab. Conclusion ①The recombinant Adenovirus with Fas gene can transfect the Fas gene into keloidderived fibroblasts and highly improved the expression of Fas protein. The newly expressed Fas gene can reconstruct the blocked Fas signal. ②Ad-Fas(B) has better therapeutic effect in vitro gene therapy. ③ The correlation between keloid and Fas gene was further proved and it may pave the way for further gene therapy in keloid .
Objective To investigate the rationale of immune privilege of testicular sertoli cell. Methods Testicular sertoli cell was prepared by digested collagenase, trypsin, and Dnase. In vitro, the sertoli cells were culture together with active lymphocytes to observe the effect on killing lymphocytes. SABC was used for labeling the Fas ligand on testicular sertoli cell.Results In vitro, sertoli cell can kill the active lymphocytes, and testicular sertoli cell expresses the Fas ligand. Conclusion Fas ligand expressing on the testicular sertoli cell may be the cause of immune privilege of testicular.
Objective Corticosteroids can destroy the cartilage. To investigate the effect of dexamethasone (Dexa) on the apoptosis and expression of Fas/FasL of human articular chondrocytes (HACs) in vitro so as to explore the mechanism ofpro-apoptotic role of Dexa on HACs. Methods Following full agreement of patients, the cartilage specimens were collectedfrom the patients with osteoarthritis undergoing knee replacement. The second passage HACs were incubated in cell culture media containing 0.125, 1.25, 12.5, 25, and 50 μg/mL Dexa for 48 hours respectively to determine the optimal concentration of Dexa by MTT. The apoptosis was assessed by TMRE/Hoechst/Annexin V-FITC/7-AAD quadruple staining after culture for 0, 24, and 48 hours. The mRNA expressions of Fas and FasL were determined by real-time quantitative PCR after culture for 48 hours. The protein expressions of Fas and FasL were determined by immunohistochemistry staining analysis after culture for 24 hours and 48 hours. Results The cell inhibitory rate of 25 μg/mL Dexa was significantly higher than that of 50 μg/mL Dexa (P lt; 0.05), and there were significant differences when compared with that at other concentrations of Dexa (P lt; 0.05), so 25 μg/mL Dexa was appropriately selected as an optimal concentration of Dexa. The apoptotic rates of HACs were 5.8% ± 0.3%, 27.0% ± 2.6%, and 36.0% ± 3.1% at 0, 24, and 48 hours, respectively, in a time dependent manner (P lt; 0.05). The expressions of Fas mRNA were (8.93 ± 1.12) × 10—3 in the experimental group and (3.31 ± 0.37) × 10—3 in the control group, showing significant difference (P lt; 0.05). The expressions of FasL mRNA were (5.92 ± 0.66) × 10—3 in the experimental group and (2.31 ± 0.35) × 10—3in the control group, showing significant difference (P lt; 0.05). The expressions of Fas and FasL proteins showed an increasing tendency with time in the experimental group and the expressions were significantly higher than those in the control group after culture for 24 hours and 48 hours (P lt; 0.05). Conclusion Dexa can induce the apoptosis and significantly upregulate the apoptotic gene expression of Fas/FasL, which can provide the experimental evidence to further investigate the role of Fas/FasL signaling pathway in Dexa-induced HACs apoptosis.