ABSTRACT
Lignans are a powerful weapon for plants to resist stresses and have diverse bioactive functions to protect human health. Elucidating the mechanisms of stereoselective biosynthesis and response to stresses of lignans is important for the guidance of plant improvement. Here, we identified the complete pathway to stereoselectively synthesize antiviral (-)-lariciresinol glucosides in Isatis indigotica roots, which consists of three-step sequential stereoselective enzymes DIR1/2, PLR, and UGT71B2. DIR1 was further identified as the key gene in respoJanuary 2024nse to stresses and was able to trigger stress defenses by mediating the elevation in lignan content. Mechanistically, the phytohormone-responsive ERF transcription factor LTF1 colocalized with DIR1 in the cell periphery of the vascular regions in mature roots and helped resist biotic and abiotic stresses by directly regulating the expression of DIR1. These systematic results suggest that DIR1 as the first common step of the lignan pathway cooperates with PLR and UGT71B2 to stereoselectively synthesize (-)-lariciresinol derived antiviral lignans in I. indigotica roots and is also a part of the LTF1-mediated regulatory network to resist stresses. In conclusion, the LTF1-DIR1 module is an ideal engineering target to improve plant Defenses while increasing the content of valuable lignans in plants.
ABSTRACT
OBJECTIVE@#AP2/ERF (APETALA2/ethylene-responsive factor) superfamily is one of the largest gene families in plants and has been reported to participate in various biological processes, such as the regulation of biosynthesis of active lignan. However, few studies have investigated the genome-wide role of the AP2/ERF superfamily in Isatis indigotica. This study establishes a complete picture of the AP2/ERF superfamily in I. indigotica and contributes valuable information for further functional characterization of IiAP2/ERF genes and supports further metabolic engineering.@*METHODS@#To identify the IiAP2/ERF superfamily genes, the AP2/ERF sequences from Arabidopsis thaliana and Brassica rapa were used as query sequences in the basic local alignment search tool. Bioinformatic analyses were conducted to investigate the protein structure, motif composition, chromosome location, phylogenetic relationship, and interaction network of the IiAP2/ERF superfamily genes. The accuracy of omics data was verified by quantitative polymerase chain reaction and heatmap analyses.@*RESULTS@#One hundred and twenty-six putative IiAP2/ERF genes in total were identified from the I. indigotica genome database in this study. By sequence alignment and phylogenetic analysis, the IiAP2/ERF genes were classified into 5 groups including AP2, ERF, DREB (dehydration-responsive element-binding factor), Soloist and RAV (related to abscisic acid insensitive 3/viviparous 1) subfamilies. Among which, 122 members were unevenly distributed across seven chromosomes. Sequence alignment showed that I. indigotica and A. thaliana had 30 pairs of orthologous genes, and we constructed their interaction network. The comprehensive analysis of gene expression pattern in different tissues suggested that these genes may play a significant role in organ growth and development of I. indigotica. Members that may regulate lignan biosynthesis in roots were also preliminarily identified. Ribonucleic acid sequencing analysis revealed that the expression of 76 IiAP2/ERF genes were up- or down-regulated under salt or drought treatment, among which, 33 IiAP2/ERF genes were regulated by both stresses.@*CONCLUSION@#This study undertook a genome-wide characterization of the AP2/ERF superfamily in I. indigotica, providing valuable information for further functional characterization of IiAP2/ERF genes and discovery of genetic targets for metabolic engineering.
Subject(s)
Abscisic Acid , Isatis/genetics , Multigene Family , Phylogeny , Homeodomain Proteins/genetics , Genome, PlantABSTRACT
OBJECTIVE@#To investigate the clinical significance and screen the risk factors of redundant nerve roots(RNRs) in patients with lumbar spinal stenosis.@*METHODS@#The clinical data of 196 patients with lumbar spinal stenosis in the department of Spinal Surgery, Yijishan Hospital, Wannan Medical College from April 1, 2015 to November 30, 2020 were retrospectively analyzed. All patients were divided into RNRs positive group and RNRs negative group according to the presence of RNRs. The differences in general clinical data, imaging parameters, visual analogue scale(VAS), Oswestry disability index(ODI), and other indicators between the two groups were compared. The risk factors which are highly correlated with RNRs were screened by binary Logistic regression analysis.@*RESULTS@#There were 59 cases in the RNRs positive group, with an occurrence rate of 29.95% (59/137), and 137 cases in the RNRs negative group. The incidence rate of RNRs in 196 patients with lumbar spinal stenosis was 30.10% (59/196). VAS and ODI scores of patients in the two groups were statistically significant (P<0.05), and clinical symptoms of patients in the RNRs positive group were more severe than those in the RNRs negative group. There were significant differences in age, number of stenosis segments, average area of lumbar dural sac, area of the narrowest segment and the narrowest segment(P<0.05). Binary logistic regression analysis showed that the number of stenosis segments, the average median sagittal diameter of spinal canal, and the average area of dural sac in lumbar intervertebral space were correlated with the generation of RNRs (P<0.05). The regression coefficient of the number of stenosis segments was -1.115, the regression coefficient of the median sagittal diameter of the spinal canal was -1.707, and the regression coefficient of the mean dural sac area of the lumbar intervertebral space was 7.556.@*CONCLUSION@#The clinical symptoms of patients with lumbar spinal stenosis accompanied by RNRs are more severe than those without them. The number of narrow segments, median sagittal diameter of the spinal canal, and the area of the lumbar intervertebral dural sac are the high-risk factors for RNRs, with the area of the lumbar intervertebral dural sac has the highest correlation.
Subject(s)
Humans , Spinal Stenosis/surgery , Constriction, Pathologic , Clinical Relevance , Retrospective Studies , Risk FactorsABSTRACT
【Objective】 To explore the causes of hypothermia in patients undergoing transurethral thulium laser prostatectomy. 【Methods】 A total of 170 patients who underwent transurethral thulium laser prostatectomy in our hospital during Sep.2020 and May 2021 were prospectively enrolled in the study. The patients were divided into normal body temperature group (n=143) and hypothermia group (n=27), based on whether perioperative hypothermia happened. The clinical data were analyzed to evaluate the risk factors of hypothermia. 【Results】 Univariate analysis showed that there were statistical differences in anesthesia time, operation time, prostate size and total amount of perfusion fluid between the two groups (P<0.05). Logistic analysis showed that the size of prostate was the independent risk factor of perioperative hypothermia (P<0.05). Patients were further divided according to prostate size. For patients with prostate < 80 g, the size of prostate was the independent risk factor of perioperative hypothermia (P<0.05), while for patients with prostate ≥ 80 g, the amount of perfusion fluid was the independent risk factor (P<0.05). 【Conclusion】 Perioperative hypothermia in patients undergoing transurethral thulium laser prostatectomy is related to the anesthesia time, operation time, prostate size and total amount of perfusion fluid. It is necessary to evaluate the risk factors before operation and take effective thermal insulation measures.
ABSTRACT
To explore the application of IL-6, PCT, T lymphocyte subsets and TIGIT expression on T lymphocytes in the evaluation of Crohn's disease status. Using a cross-sectional study, total of 119 confirmed patients with Crohn's disease who were treated in the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine from June 2020 to December 2022 were selected. The age range was 18-59 years old, and the median age (interquartile range) was 37 (29, 45) years old, including 57 cases in active disease group (30 males, 27 females), 62 cases in disease remission group (33 males, 29 females); 50 healthy control groups (27 males, 23 females), the age range was 19-60 years old, and the median age (interquartile range) was 38 (31, 46) years old. The level of IL-6 was detected by flow fluorescence microsphere method, the concentration of PCT was detected by immunochromatography, and the levels of T lymphocyte subsets and TIGIT were detected by flow cytometry. The differences and correlations between the detection indicators in each group were compared, logistic regression was used to analyze the factors influencing the progression of Crosne's disease and the clinical value of each detection indicator was analyzed by ROC curve. The results showed that there were no statistically significant differences in age and gender among the control group, the remission group, and the active group (H=1.422,χ2=0.020;P=0.491, P=0.990); in the active group, IL-6 was 17.55(9.67, 21.72)pg/ml, PCT was 0.38(0.14, 0.43)ng/ml, CD3+CD4+ was 35.47%±6.01%, CD3+CD8+ was 30.50%±5.20%, TIGIT was 25.08%±6.30%; in the remission group, IL-6 was 8.46(5.21, 10.04) pg/ml, PCT was 0.26(0.11, 0.35) ng/ml, CD3+CD4+ was 37.62%±4.86%, CD3+CD8+ was 28.30%±5.28%, TIGIT was 34.22%±5.45%; in the control group, IL-6 was 6.13(3.57, 8.12)pg/ml, PCT was 0.17(0.10, 0.21)ng/m, CD3+CD4+ was 39.74%±3.94%, CD3+CD8+ was 26.59%±4.50%, and TIGIT was 37.64%±6.22%.There were significant differences in IL-6, PCT, CD3+CD4+%, CD3+CD8+%, and TIGIT among the three groups(H=58.688, H=18.003, F=9.600, F=8.124, F=65.059;P<0.001, P<0.001, P<0.001, P<0.001, P<0.001), Among them, IL-6 and TIGIT in the active group were significantly different from those in the remission group (P<0.001, P<0.001), and only TIGIT was significantly different between the remission group and the control group (P=0.007);Spearman correlation analysis showed that the expression of TIGIT on T lymphocytes was negatively correlated with the levels of IL-6; the results of Logistic regression analysis showed that IL-6, PCT and TIGIT were independent factors affecting the progression of Crohn's disease;Comparing the ROC curves of the active group and the remission group, found that TIGIT was significantly different from PCT, CD3+CD4+, CD3+CD8+(Z=4.011, Z=4.091, Z=4.157; P<0.001, P<0.001, P<0.001), no statistical difference with IL-6 (Z=1.193, P=0.233). Selected the combined detection of IL-6 and TIGIT with the best AUC area and Youden index, which shows that the clinical value is improved, the AUC area of IL-6+TIGIT was significantly different from that of IL-6 (Z=2.674, P=0.008). In summary, TIGIT of T lymphocytes and IL-6 detection may be valuable in the diagnosis and treatment of Crohn's disease, and the combined detection of TIGIT and IL-6 may be meaningful for evaluating the status of Crohn's disease.
Subject(s)
Male , Female , Humans , Adult , Adolescent , Young Adult , Middle Aged , Interleukin-6 , Crohn Disease/diagnosis , Cross-Sectional Studies , T-Lymphocyte Subsets , Receptors, ImmunologicABSTRACT
To explore the application of IL-6, PCT, T lymphocyte subsets and TIGIT expression on T lymphocytes in the evaluation of Crohn's disease status. Using a cross-sectional study, total of 119 confirmed patients with Crohn's disease who were treated in the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine from June 2020 to December 2022 were selected. The age range was 18-59 years old, and the median age (interquartile range) was 37 (29, 45) years old, including 57 cases in active disease group (30 males, 27 females), 62 cases in disease remission group (33 males, 29 females); 50 healthy control groups (27 males, 23 females), the age range was 19-60 years old, and the median age (interquartile range) was 38 (31, 46) years old. The level of IL-6 was detected by flow fluorescence microsphere method, the concentration of PCT was detected by immunochromatography, and the levels of T lymphocyte subsets and TIGIT were detected by flow cytometry. The differences and correlations between the detection indicators in each group were compared, logistic regression was used to analyze the factors influencing the progression of Crosne's disease and the clinical value of each detection indicator was analyzed by ROC curve. The results showed that there were no statistically significant differences in age and gender among the control group, the remission group, and the active group (H=1.422,χ2=0.020;P=0.491, P=0.990); in the active group, IL-6 was 17.55(9.67, 21.72)pg/ml, PCT was 0.38(0.14, 0.43)ng/ml, CD3+CD4+ was 35.47%±6.01%, CD3+CD8+ was 30.50%±5.20%, TIGIT was 25.08%±6.30%; in the remission group, IL-6 was 8.46(5.21, 10.04) pg/ml, PCT was 0.26(0.11, 0.35) ng/ml, CD3+CD4+ was 37.62%±4.86%, CD3+CD8+ was 28.30%±5.28%, TIGIT was 34.22%±5.45%; in the control group, IL-6 was 6.13(3.57, 8.12)pg/ml, PCT was 0.17(0.10, 0.21)ng/m, CD3+CD4+ was 39.74%±3.94%, CD3+CD8+ was 26.59%±4.50%, and TIGIT was 37.64%±6.22%.There were significant differences in IL-6, PCT, CD3+CD4+%, CD3+CD8+%, and TIGIT among the three groups(H=58.688, H=18.003, F=9.600, F=8.124, F=65.059;P<0.001, P<0.001, P<0.001, P<0.001, P<0.001), Among them, IL-6 and TIGIT in the active group were significantly different from those in the remission group (P<0.001, P<0.001), and only TIGIT was significantly different between the remission group and the control group (P=0.007);Spearman correlation analysis showed that the expression of TIGIT on T lymphocytes was negatively correlated with the levels of IL-6; the results of Logistic regression analysis showed that IL-6, PCT and TIGIT were independent factors affecting the progression of Crohn's disease;Comparing the ROC curves of the active group and the remission group, found that TIGIT was significantly different from PCT, CD3+CD4+, CD3+CD8+(Z=4.011, Z=4.091, Z=4.157; P<0.001, P<0.001, P<0.001), no statistical difference with IL-6 (Z=1.193, P=0.233). Selected the combined detection of IL-6 and TIGIT with the best AUC area and Youden index, which shows that the clinical value is improved, the AUC area of IL-6+TIGIT was significantly different from that of IL-6 (Z=2.674, P=0.008). In summary, TIGIT of T lymphocytes and IL-6 detection may be valuable in the diagnosis and treatment of Crohn's disease, and the combined detection of TIGIT and IL-6 may be meaningful for evaluating the status of Crohn's disease.
Subject(s)
Male , Female , Humans , Adult , Adolescent , Young Adult , Middle Aged , Interleukin-6 , Crohn Disease/diagnosis , Cross-Sectional Studies , T-Lymphocyte Subsets , Receptors, ImmunologicABSTRACT
@# Objective - ( ) To explore the influence on the diagnosis of occupational noise induced deafness ONID using three , Methods versions of diagnostic criteria in 2002 2007 and 2014. A total of 1 766 workers who asked for ONID diagnosis were selected as the research subjects using judgment sampling method. The results of pure tone audiometry were collected. GBZ 49-2002Diagnostic Criteria of Occupational Noise-inducedHearing Loss( The ONID was diagnosed using hereinafter referred to as GBZ 49-2002),GBZ 49-2007Diagnostic Criteria of Occupational Noise-induced Deafness( GBZ 49-2007) hereinafter referred to as GBZ 49-2014 Diagnostic of Occupational Noise-induced Deafness( GBZ 49-2014), and hereinafter referred to as and the Results - - , - diagnostic results were compared. Compared with GBZ 49 2002 and GBZ 49 2007 diagnosis with GBZ 49 2014 had ( vs , vs , P ), ( vs , a higher rate of ONID 57.9% 66.0% 44.8% 66.0% both <0.01 and had a higher rate of mild ONID 47.3% 54.6% vs , P ) - - 36.0% 54.6% both <0.01 . The diagnostic rate for ONID using GBZ 492014 was higher than those using GBZ 49 2002 and - ( P )Conclusion - GBZ 49 2007 in each age groups all <0.01 . GBZ 49 2014 improved the diagnostic rate of ONID compared - - with GBZ 49 2002 and GBZ 49 2007. The reason is related to the inclusion of 4 000 Hz hearing threshold with a weight of 0.1 - as the diagnostic hearing threshold and the use of a new age and gender correction method in GBZ 49 2014.
ABSTRACT
This paper reviews the application of "co-regulation of body and mind" of acupuncture for post-stroke spasticity. It is found that acupoints on the head and the back of the governor vessel, as well as Jiaji (Ex-B 2) points are mainly used for regulating the mind, and the local sites of spastic muscles and the points on the antagonistic muscles are for regulating the body specially. It is believed that regulating the mind should be integrated with regulating the body, while, the acupoint selection be associated with needling methods so as to fully achieve the "co-regulation of body and mind" and enhance the practical value of acupuncture for post-stroke spasticity. It is proposed that the classical anti-spastic needling techniques, such as huici (relaxing needling) and guanci (joint needling), should be more considered.
Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy/methods , Muscle Spasticity/therapy , Muscles , Stroke/therapyABSTRACT
Cannulated Screw is a common internal fixation for the treatment of femoral neck fractures. However, the traditional implantation method has disadvantages such as inaccuracy and large radiation exposure. Based on the anatomical characteristics of the femoral neck and geometric principles, we develop a novel guide device for cannulated screws insertion. The cadaver experiment showed that it can improve the accuracy of cannulated screws implantation, reduce puncture attempts and the radiation exposure of doctors and patients.
Subject(s)
Humans , Bone Screws , Femoral Neck Fractures/surgery , Fracture Fixation, Internal , Robotic Surgical ProceduresABSTRACT
@#[摘 要] 目的:设计并制备一种分别靶向B细胞表面抗原CD19和CD22的CAR-T细胞,检测其对肿瘤细胞的体内外杀伤效果。方法:将含有人源化 CD19 ScFv的二代CAR分子和带有CD3ε链作为共刺激结构域的CD22 ScFv CAR分子以P2A自剪切肽连接,序列连接于慢病毒载体pLTR-CMV-MCS中,以HEK-293T细胞包装相应的慢病毒载体,感染健康志愿者提供的T细胞制备CAR-19-22-T细胞,同时以相同二代结构分别构建单靶向CAR-T细胞作为参照。构建表达荧光素酶、CD19和/或CD22的前列腺癌3M细胞(靶细胞)。将各种CAR-T细胞与靶细胞共同培养,采用荧光素酶化学发光法和ELISA法检测其对靶细胞的杀伤能力和细胞因子的分泌水平。通过尾静脉注射Raji-Luc细胞构建NOD-SCID免疫缺陷小鼠白血病模型,分别注射各组CAR-T细胞进行治疗并评估其疗效。结果:培养7 d的CAR-19-22-T细胞的CAR-19表达率为13.7%,CAR-22表达率为14.3%。CAR-19-22-T细胞在10∶1效靶比时,对3M-CD19-Luc、3M-CD22-Luc和3M-CD19-CD22-Luc细胞的杀伤率均显著高于T细胞[(78.1±14.4)% vs (11.1±4.3)%、(46.7±10.7)% vs (12.4±2.7)%、(90.5±4.3)% vs (14.3±3.7)%,均P<0.01];与3M-CD19-Luc、3M-CD22-Luc、3M-CD19-CD22-Luc靶细胞共培养后,CAR-19-22-T细胞IFN-γ、TNF-α和IL-2水平均显著低于CAR-19-T和CAR-22-T细胞(P<0.05或P<0.01)。CAR-19-22-T细胞对移植Raji-Luc细胞模型小鼠治疗效果明显,其生存期显著长于T细胞组(P<0.01),与CAR-19-T组和CAR-22-T组荷瘤小鼠比较差异均无统计学意义(均P>0.05)。结论:成功设计并制备了一种双靶点CAR-19-22-T细胞,其能够有效杀伤表达CD19和/或CD22抗原的肿瘤细胞,对Raji-Luc细胞的白血病模型小鼠有显著的治疗效果。
ABSTRACT
Salvia miltiorrhiza is widely used in the treatment of the angina pectoris, coronary heart disease and myocardial infarction. CYP76AH3 is the key P450 enzyme, locating at the branch point of the tanshinone synthesis pathway. The crystal structural study and key amino acid analysis are of great significance for synthetic biology study on tanshinone. But it is always a challenge for scientists to carry out protein purification, crystallization and crystal structural studies on transmembrane type Ⅱ P450 enzymes. In this study, the prokaryotic expression plasmid was generated, and the high-purity target protein was purified. CYP76AH3 was successfully crystallized, and the crystal structure was solved.After docking range was determined by Cavityplus analysis, molecular docking with Discovery Studio was conducted. The docking result indicated that Gly298 and Asp294 had hydrogen bond interaction with the substrate, while Phe479, Leu367 and Leu293 had hydrophobic interaction with the substrate.In addition, the effect of mutations at the key amino acids on the protein structure stability was predicted throung point mutation simulation. This study would provide a target for protein engineering of CYP76AH3 and lay a foundation for the study of synthesis biology on tanshinones.
ABSTRACT
OBJECTIVE@#To investigate the relationship between spine-pelvic sagittal parameters and clinical efficacy before and after oblique lumbar interbody fusion(OLIF).@*METHODS@#A retrospective analysis of clinical data of 65 patients with lumbar degenerative diseases treated with OLIF were performed from July 2017 to July 2018. There were 26 males and 39 females aged from 33 to 79 years old with an average of (62.72±10.23) years old. Oswestry Disability Index (ODI) and visual analogue scale (VAS) before and at the latest follow up were evaluated. Disc height (DH) and spine- pelvic sagittal parameters of the surgical segment were measured before and at the latest follow- up, including pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS) and lumbar lordosis (LL). According to the difference of PI-LL, it was judged whether PI and LL match and the patients were grouped, PI-LL ranged from -9° to 9° was set as matching group, and PI-LL less than -9° or larger than 9° was set as mismatching group. The spine-pelvic sagittal parameters were analyzed before and at the latest follow-up of OLIF in patients with lumbar degenerative diseases, and the correlation between changes and clinical efficacy was compared.@*RESULTS@#All patients were followed up from 8 to 20 months with an average of (14.20±3.68) months. Operation time was (91.54±25.97) min, intraoperative blood loss was (48.15±10.14) ml, and the hospitalization time ranged from 6 to 19 days with an average of (9.28± 2.50) days. Totally 84 surgical levels, 46 patients were single segment and 19 patients were double segments. VAS and ODI score were improved from (4.88±0.99) point, (67.60±13.73) % preoperatively to (2.85±1.30) points, (30.57±6.48) % at the latest follow-up. There were significant differences in VAS and ODI scores between before and at the latest follow-up. The sagittal parameters of LL, PT, SS, PI, PI -LL and the surgical level DH were (42.80 ±16.35)° , (23.22 ±10.91)° , (26.95 ± 13.30)°, (50.22±14.51)°, (7.53±16.13) °, (0.91±0.29) cm preoperatively and improved to the latest follow-up (49.95± 12.82) °, (17.94±9.24) °, (33.71±12.66) °, (51.65±10.26) °, (1.68±17.00) °, (1.20±0.40) cm;there were statistical differences in LL, PT, SS, PI-LL, DH before operation and at the latest follow up, while no difference in PI. LL of preoperative PI-LL in matched group was (48.76±11.09)° , and (38.00±18.37)° in PI-LL mismatch group, there was difference between two groups. There were no differences in VAS, ODI, PT, SS, PI and DH between two groups. At the latest follow-up, ODI between PI-LL matched group and PI-LL mismatched group were (29.40±5.93)% and (32.86±7.02)% respectively, and had difference in ODI between two groups;while there were no significant differences in VAS, LL, PT, SS, PI, and DH. Pearson correlation analysis showed preoperative PT-LL was positively correlated with VAS;PT was positively correlated with ODI at the latest follow-up.@*CONCLUSION@#OLIF has a good surgical effect on lumbar degenerative diseases, and could change spine-pelvic sagittal parameters of patient to a certain extent, and further restoring the balance of the sagittal plane of lumbar spine.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Lumbar Vertebrae , Lumbosacral Region , Pelvis , Retrospective Studies , Spinal Fusion , Treatment OutcomeABSTRACT
AP2/ERF transcription factor is a kind of transcription factors widely existing in plants, and contains at least a conserved AP2/ERF domain composed of about 60-70 amino acids. AP2/ERF transcription factors are widely involved in a variety of physiological processes in plants, including plant development, fruit ripening, flower development and other plant development processes, as well as such stress response processes as damage, pathogen defense, high-salt condition and drought. In recent years, secondary metabolic engineering that takes transcription factors as genetic manipulation targets has developed rapidly in improving the content of active ingredients and the quality of medicinal plants. This paper reviews the recent progress in the regulation of secondary metabolites biosynthesis with AP2/ERF transcription factors, and provides theoretical basis for the exploration of efficient regulatory targets, the regulation of secondary metabolites in medicinal plants, the targeted improvement of the content of active ingredients in traditional Chinese medicine, and the sustainable supply of high-quality traditional Chinese medicines.
Subject(s)
Gene Expression Regulation, Plant , Phylogeny , Plant Proteins/metabolism , Transcription Factor AP-2/metabolism , Transcription Factors/metabolismABSTRACT
Objective@#To investigate the expression of inhibitory receptor TIGIT gene in peripheral NK cells of patients with rheumatoid arthritis (RA) and its clinical significance.@*Methods@#A case control study was conducted of 58 RA patients(30 patients with active RA disease, 28 patients with remission of RA) and 22 healthy controls (HC) in the department of rheumatology and immunology from Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine during December 2018 to June 2019, the related clinical data were collected. Flow cytometry was used to compare the expression of TIGIT gene on peripheral NK cells. The IFN-γ secretion level of cytokines in peripheral blood was detected by ELISA, and analyzed the correlations between TIGIT gene and disease activity and IFN-γ level. T-test or non-parametric test was used for comparison between the two groups, and Pearson correlation analysis was used for correlation between the two variables.@*Results@#Compared with HC group, the number of NK cells in RA disease group was reduced, which was (13.88±4.56) ×107 cells/L in RA disease group and (25.69± 2.48) ×107 cells/L in HC group (t=-2.036, P=0.041). The percentage of TIGIT gene expression in peripheral blood NK cells was not statistically different between RA disease group (39.73±9.37)% and HC group (45.64±9.91)% (t=-1.241, P=0.218). However, the average fluorescence intensity (MFI) of TIGIT gene expression was decreased, which was (7.21±2.03) in RA group and (9.01±3.29) in HC group (t=-2.947, P=0.004).MFI of TIGIT gene in NK cells of the disease active subgroup was (6.72±2.01), lower than that of the disease remission subgroup (8.75±2.64), (t=-3.316, P=0.002), and MFI of TIGIT gene in NK cells of the disease active subgroup was negatively correlated with DAS28 score (r2=0.649 6, P<0.000 1).The secretion level of IFN-γ cytokine in the RA disease group was (67.13±14.84) pg/ml, higher than that in the HC group (57.21±14.23) pg/ml (t=2.757, P=0.017), and the secretion level of IFN-γ cytokine in the disease active subgroup was negatively correlated with the MFI of TIGIT gene on NK cells (r2=0.662 2, P<0.000 1). Experimental results of peripheral blood mononuclear cell stimulation in the RA disease activity subgroup showed that the secretion level of cytokines IFN-γ was reduced after stimulation compared with that before stimulation (t=11.38, P<0.000 1).@*Conclusion@#The abnormal expression of TIGIT gene on peripheral NK cells are observed in patients with RA, which correlate with disease activity and IFN-γ secretion level.
ABSTRACT
The filiform needling technique is an important factor affecting the acupoint effect, and it is the key to option the needling technique corresponding to the disease so that the clinical curative effect can be improved. This paper systematically reviews the application of kinetic needling in the treatment of spasm, in order to provide some theoretical basis for the optimal acupuncture regimen of spasm. By summarizing and analyzing the similarities and differences of acupoint selection principle, needling characteristics, stimulation range, stimulation amount and indications in the treatment of spasm, it is found that kinetic needling emphasizes the effective combination of acupuncture and kinesis, which is an effective mean of treating spasm.
Subject(s)
Humans , Acupuncture Points , Spasm , Vascular Surgical ProceduresABSTRACT
Objective To investigate the expression of inhibitory receptor TIGIT gene in peripheral NK cells of patients with rheumatoid arthritis (RA) and its clinical significance. Methods A case control study was conducted of 58 RA patients(30 patients with active RA disease, 28 patients with remission of RA) and 22 healthy controls (HC) in the department of rheumatology and immunology from Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine during December 2018 to June 2019, the related clinical data were collected. Flow cytometry was used to compare the expression of TIGIT gene on peripheral NK cells. The IFN-γ secretion level of cytokines in peripheral blood was detected by ELISA, and analyzed the correlations between TIGIT gene and disease activity and IFN-γ level. T-test or non-parametric test was used for comparison between the two groups, and Pearson correlation analysis was used for correlation between the two variables. Results Compared with HC group, the number of NK cells in RA disease group was reduced, which was (13.88 ± 4.56) × 107 cells/L in RA disease group and (25.69 ±2.48) ×107 cells/L in HC group (t=-2.036, P=0.041). The percentage of TIGIT gene expression in peripheral blood NK cells was not statistically different between RA disease group (39.73 ± 9.37) % and HC group (45.64 ± 9.91) % (t=-1.241, P=0.218). However, the average fluorescence intensity (MFI) of TIGIT gene expression was decreased, which was (7.21±2.03) in RA group and (9.01±3.29) in HC group (t=-2.947,P=0.004).MFI of TIGIT gene in NK cells of the disease active subgroup was (6.72±2.01), lower than that of the disease remission subgroup (8.75 ± 2.64), (t=-3.316, P=0.002), and MFI of TIGIT gene in NK cells of the disease active subgroup was negatively correlated with DAS28 score (r2=0.6496, P<0.0001).The secretion level of IFN-γcytokine in the RA disease group was (67.13±14.84) pg/ml, higher than that in the HC group (57.21 ± 14.23) pg/ml (t=2.757, P=0.017), and the secretion level of IFN-γ cytokine in the disease active subgroup was negatively correlated with the MFI of TIGIT gene on NK cells (r2=0.6622, P<0.0001). Experimental results of peripheral blood mononuclear cell stimulation in the RA disease activity subgroup showed that the secretion level of cytokines IFN-γwas reduced after stimulation compared with that before stimulation (t=11.38, P<0.0001). Conclusion The abnormal expression of TIGIT gene on peripheral NK cells are observed in patients with RA, which correlate with disease activity and IFN-γsecretion level.
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OBJECTIVE@#To investigate the effects of mangiferin on proliferation, apoptosis and cycle of FLT3-ITD mutation-positive acute myeloid leukemia cells and its mechanism.@*METHODS@#The effects of different concentration of mangiferin on proliferation of MV4-11 cells were detected by CCK8 method. Apoptosis, cell cycle and FLT3 transmembrane protein expression were detected by flow cytometry. FLT3 mRNA expression was detected by real-time fluorescent quantitative polymerase chain reaction (PCR) .@*RESULTS@#Mangiferin obviously inhibited MV4-11 proliferation in a concentration and time dependent manner (48 h,r=0.922;72 h,r=0.959;96 h,r=0.973). The ratio of G0/G1 phase in cell cycle increased with the enhancement of concentration of mangiferin in MV4-11 cells for 48 h, and the ratio of S phase decreased with enhasment of concentration. The increase of apoptosis was more obvious. The expression of FLT3 transmembrane protein significantly decreased after the actior of IC50 concentration of mangiferin in MV4-11 cells for 48 h. The results of qRT-PCR showed that the expression of FLT3 mRNA significantly decreased after treatment of MN4-11 cells with mangiferin (P<0.05).@*CONCLUSION@#Mangiferin inhibits MV4-11 cell proliferation, arrests cell cycle progression and promotes apoptosis, which may be related with the inhibition of FLT3 activity by mangiferin and the subsequent signaling pathways involved in apoptosis and proliferation of cells.
Subject(s)
Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Leukemia, Myeloid, Acute , Mutation , Xanthones , fms-Like Tyrosine Kinase 3ABSTRACT
Abstract This study aimed to determine the association between the polymorphisms and haplotypes in the xeroderma pigmentosum group D (XPD) gene and the risk of pancreatic cancer in the Chinese Han population. SNaPshot was used for genotyping six SNP sites of the XPD gene. Comparisons of the correlations between different genotypes in combination with smoking and the susceptibility to pancreatic cancer were performed. Individual pancreatic cancer risk in patients who carry mutant C alleles (AC, CC, and AC+CC) at rs13181 increased (p < 0.05). Taking non-smoking individuals who carry the AA genotype as a reference, and non-smoking individuals who carry mutant allele C (AC+CC), the risk of pancreatic cancer increased by 3.343 times in individuals who smoked ≥ 20 cigarettes daily, 3.309 times in individuals who smoked ≥ 14 packs per year, 5.011 times in individuals who smoked ≥ 24 packs per year, and 4.013 times in the individuals who smoked ≥ 37 packs per year (P < 0.05). In addition, haplotype analysis revealed that haplotype AGG, which comprised rs13181, rs3916874 and rs238415, was associated with a 1.401-fold increase in pancreatic cancer risk (p < 0.05). We conclude that the polymorphism of XPD Lys751Gln (rs13181) in combination with smoking contributes to increased risk of pancreatic cancer in the Chinese Han population. Haplotype AGG might be a susceptibility haplotype for pancreatic cancer.
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BACKGROUND: ClonePix, a cell cloning and screening system, can quickly and efficiently screen cell clones undergoing suspension culture with low melting point agarose, which can be used to provide a sufficient number of seed cells for biological therapies. OBJECTIVE: To isolate and identify chondrogenic stem cells from the rat lumbar endplate.METHODS: The lumbar endplates of 10 Sprague-Dawley rats with an age of 4 weeks were digested using trypsin and type II collagenase to isolate primary chondrogenic stem cells, followed by suspension culture with low melting point agarose. Then, the cell clones were selected by ClonePix and expanded in vitro for morphological observation.The multidirectional differential potential of cell clones was identified through osteogenesis, adipogenesis and chondrogensis tests, and the monoclonal formation ability was determined. RESULTS AND CONCLUSION: The cell clones could be successfully separated by using the agarose culture system, which were spindle-shaped after in vitro expansion.The osteogenesis,adipogenesis and chondrogensis capacities of the cells were identified using alizarin red, oil red O and safranin staining, respectively. Single cells of the clone group were inoculated into 10 cm culture dishes, and after 12 days of in vitro expansion culture,colonies of cells were observed with the naked eye.With the increase of cell seeding density,the number of cell colonies decreased. When the cells were inoculated into 10 cm culture dishes at a density of 100 cells, the colony forming ability was strongest, and more than 20 cell colonies could be formed. These findings indicate that chondrogenic stem cells that are isolated by the agarose gel culture system have multidirectional differentiation potential and high proliferation ability.
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Objective?To analyze the clinical efficacy and related X-ray findings of patients underwent arthroscopic treatment of femoral acetabular impingement (FAI) syndrome with different anatomical features.?Methods?Twenty-four patients with FAI underwent arthroscopic surgery from September 2015 to December 2016 were selected to analyze the clinical features, postoperative pain, knee joint function, activity and complications.?Results?Compared with those before treatment, the visual analogue scale (VAS) scores of patients at 7 d, 1 month, 3 months and 6 months after treatment were significantly lower than those before treatment, while the Harris scores were significantly increased, at the same time patients’ knee activity was significantly increased, The difference was statistically significant (P < 0.05). The α angle of the hip joint of the cam-type patient was significantly higher than that of the jaw-type, while the eccentricity, acetabular depth, acetabular coverage and centerline (CE) angle were significantly lower than the jaw-type, and the difference was statistically significant (P < 0.05); The α angle of the cam-type patient was significantly higher than that of the healthy person’s hip joint, and the eccentricity was significantly lower than that of the healthy person’s hip joint. The difference was statistically significant (P < 0.05); the acetabular depth and hip of the clamp-type patient Radon coverage and CE angle were higher than those of hip joints in healthy people, and the difference was statistically significant (P < 0.05). There was no significant difference in acetabular anterior tilt between the three groups (P > 0.05). The incidence of complications in 24 patients underwent arthroscopy was 20.83%.?Conclusion?Hip arthroscopic treatment of hip impingement syndrome can shorten the patient’s pain relief, improve knee function and activity, its effect is good, and different hip anatomical X-ray film was significantly different.