ABSTRACT
Objective:To detect the levels of vascular endothelial growth factor (VEGF) and nitric oxide (NO) in the serum of patients with early diabetic retinopathy (DR) and to explore their value in the treatment of DR.Methods:Fifty-nine patients with type 2 diabetes mellitus from March 2019 to June 2020 in the Yulin Second Hospital were collected and divided into the non-diabetic retinopathy (NDR) group (32 cases) and background stage diabetic retinopathy (BDR) group (27 cases) according to the DR grading criteria, and 29 healthy subjects were enrolled in the healthy control (HC) group. Serum VEGF levels were measured by a double antibody sandwich enzyme-linked immunosorbent assay (ELISA), and serum NO levels were measured by the nitrate reductase method (one-step method). Pearson analysis was used to correlate serum VEGF with NO levels in each group, and a logistic regression model was used to analyze the risk factors for DR.Results:Compared with the HC group, the differences in serum VEGF and NO levels of patients in the BDR group were statistically significant (all P < 0.05). The serum VEGF and NO levels were positively correlated in all groups (all P < 0.01). Duration of diabetes, smoking, glycated hemoglobin A1c (HbA1c), serum VEGF, and NO levels were high risk factors for DR. Conclusions:Serum VEGF and NO levels may be potential pathological mechanisms for the development of microvascular complications in patients with type 2 diabetes, and the interaction may induce or accelerate the progression of DR. This study is expected to provide a new idea of complementary or combined treatment in the treatment of DR.
ABSTRACT
Objective:To investigate the changes of intestinal microecology in the early stage of sepsis rat model by 16S rDNA sequencing.Methods:Sixty male Sprague-Dawley (SD) rats were randomly divided into cecal ligation and puncture (CLP) group and sham operation group (Sham group), with 30 rats in each group. In the CLP group, sepsis rat model was reproduced by CLP method; the rats in the Sham group only underwent laparotomy without CLP. At 24 hours after the operation, the intestinal feces and serum samples of 8 rats in each group were collected. The survival rate of the rest rats was observed until the 7th day. The level of serum tumor necrosis factor-α (TNF-α) was detected by enzyme-linked immunosorbent assay (ELISA). Intestinal feces were sequenced by 16S rDNA sequencing technology. The operational taxonomic unit (OTU) data obtained after sequence comparison and clustering was used for α diversity and β diversity analysis, principal coordinate analysis and linear discriminant analysis effect size analysis (LEfSe) to observe the changes of intestinal microecology in early sepsis rats and excavate the marker flora.Results:At 24 hours after the reproduction of the model, the rats in the CLP group showed shortness of breath, scattered hair and other manifestations, and the level of serum TNF-α increased significantly as compared with that in the Sham group (ng/L: 43.95±9.05 vs. 11.08±3.27, P < 0.01). On the 7th day after modeling, the cumulative survival rate of the Sham group was 100%, while that of the CLP group was 31.82%. Diversity analysis showed that there was no significant difference in α diversity parameter between the Sham group and the CLP group (number of species: 520.00±52.15 vs. 492.25±86.61, Chao1 richness estimator: 707.25±65.69 vs. 668.93±96.50, Shannon index: 5.74±0.42 vs. 5.79±0.91, Simpson index: 0.93±0.03 vs. 0.94±0.05, all P > 0.05). However, the β diversity analysis showed that the difference between groups was greater than that within groups whether weighted according to OTU or not (abundance weighted matrix: R = 0.23, P = 0.04; abundance unweighted matrix: R = 0.32, P = 0.01). At the phylum level, the abundance of Proteobacteria and Candidatus_sacchari in the CLP group increased significantly as compared with the Sham group [18.100% (15.271%, 26.665%) vs. 6.974% (2.854%, 9.764%), 0.125% (0.027%, 0.159%)% vs. 0.018% (0.008%, 0.021%), both P < 0.05]. At the genus level, the abundance of opportunistic pathogen including Helicobacter, Ruthenium, Streptococcus, Clostridium ⅩⅧ in the CLP group was significantly higher than that in the Sham group [5.090% (1.812%, 6.598%) vs. 0.083% (0.034%, 0.198%), 0.244% (0.116%, 0.330%) vs. 0.016% (0.008%, 0.029%), 0.006% (0.003%, 0.010%) vs. 0.001% (0%, 0.003%), 0.094% (0.035%, 0.430%) vs. 0.007% (0.003%, 0.030%), all P < 0.05], and the abundance of probiotics such as Alloprevotella and Romboustia was significantly lower than that in the Sham group [7.345% (3.662%, 11.546%) vs. 22.504% (14.403%, 26.928%), 0.113% (0.047%, 0.196%) vs. 1.229% (0.809%, 2.29%), both P < 0.01]. LEfSe analysis showed that the probiotics belonging to Firmicutes were significantly enriched in the Sham group, and Romboustia was the most significantly enriched species. Opportunistic pathogens such as Helicobacter, Streptococcus and Clostridium ⅩⅧ were significantly enriched in the CLP group, Helicobacter_NGSU_ 2015 was the most significantly enriched species. Conclusion:In the early stage of sepsis, the intestinal microbiota structure of rats is significantly changed, which mainly shows that the abundance of Alloprevotella and other probiotics is significantly reduced, while that of Helicobacter and other opportunistic pathogens is significantly increased.
ABSTRACT
Objective To investigate the clinical significance of peripheral blood Th17 cells and their cytokines in the patients with tongue squamous cell carcinoma.Methods Sixty-six cases of tongue squamous cell carcinoma in our hospital from January 2013 to January 2016 served as the research subjects and contemporaneous 42 volunteers undergoing healthy physical examination were selected as the control group.Peripheral blood was collected in all subjects.The proportion of peripheral blood Th17 cells was detected by flow cytometry.The IL-17,TGF-β and IL-6 cytokine levels were detected by ELISA.Results The proportion of peripheral blood Th17 cells,IL-17,TGF-β and IL-6 in the observation group and the control group were (1.46±0.41)% vs.(0.31±0.12)%,(123.36±21.20)pg/mL vs.(20.76±8.95)pg/mL,(215.80±21.52)pg/mL vs.(26.90±10.41)pg/mL,(17.32±8.02)pg/mL vs.(5.85±1.49)pg/mL respectively,the differences were statistically significant(P<0.05).The proportion of peripheral blood Th17 cells,IL-17,TGF-β and IL-6 levels were increased with the increase of tongue squamous cell carcinoma clinical stage (P<0.01).The TGF-β level was positively correlated with the IL-17 level (r=0.626,P=0.021),the IL-6 level was positively correlated with the proportion of Th17 cells and IL-17 (r=0.626,0.597,P=0.021,0.034).Conclusion The increase of Th17 cells and IL-17 expression plays an important role in the development and progression of tongue squamous cell carcinoma,Th17 cells and IL-17 cytokine can become the important target spots of anti-tumor therapy.
ABSTRACT
This article was aimed to study the mechanism of Liu-Wei Bu-Qi (LWBQ) Capsules among rat models of chronic obstructive pulmonary disease (COPD) accompanied with reduced lung function based on MMPs/TIMPs and Th1/Th2. A total of 75 rats were randomly divided into the normal group, model group, LWBQ group, Jin-Shui-Bao (JSB) group, spleen aminopeptidase group. Except the normal group, smoke plus lipopolysaccharide tracheal instil-lation method was applied among rats in other groups to establish COPD rat model. Medication was given on the 28th day after the model was established. The medication was given for 30 days. Observation was given on changes of lung histology, lung function, interleukin (IL)-4, interferon-γ (IFN-γ), matrix metalloproteinases (MMP-9) and MMPs inhibitor 1(TIMP-1). The results showed that compared with the normal group, in the model group, lung tis-sues was damaged, and lung function was obviously reduced, while the level of inflammatory factor such as IL-1β, IFN-γ, Th1/Th2 were obvious increased (P< 0.05 or P< 0.01); while inflammation-inhibiting factors such as IL-4 and IL-35 were obviously decreased (P < 0.05 or P < 0.01); MMP-9 gene and protein expression of lung tissues were obviously increased (P< 0.05 or P< 0.01); TIMP-1 gene and protein expression were obviously decreased (P< 0.05 or P < 0.01). After medication, compared with the model group, in the LWBQ group, IFN-γ and Th1/Th2 were obviously decreased (P< 0.05 or P< 0.01); MMP-9 gene and protein expression were obviously decreased (P< 0.05 or P < 0.01); TIMP-1 expression was obviously increased (P < 0.05 or P < 0.01). Compared with the JSB group and spleen aminopeptidase group, the lung function and TIMP1 gene protein expression were increased, while MMP-9 and Th1/Th2 expression were obviously decreased (P < 0.05). It was concluded that LWBQ Capsules up-regulate IL-4 and TIMP-1, downregulate IFN-γ, Th1/ Th2 and MMP-9 expression, in order to reduce inflammatory response and improve lung function among COPD cases.
ABSTRACT
Objective To observe the changes in forkhead/winged helix transcription factor p3(Foxp3), regulatory T cells(Treg),retinoid-related orphan receptor gamma(RORγt)in rat model of chronic obstructive pulmonary disease(COPD). Methods Twenty Sprague-Dawley(SD)rats were randomly divided into normal control group and COPD model group,with 10 rats in each group. The COPD model was reproduced by smoke inhalation and tracheal instillation of lipopolysaccharide(LPS),and no such treatment was conducted in normal control group. Twenty-eight days after the model reproduction,the pulmonary function was determined,the pathological changes of lung tissue were observed with haematoxylin-eosin(HE)staining,interleukins(IL-6,IL-10)in serum were detected by enzyme-linked immunosorbent assay(ELISA),CD4+CD25+Foxp3+Treg of peripheral blood was determined by flow cytometry,and the expressions of Foxp3,RORγt,IL-17 protein in lung tissue were assayed by Western Blot. Results Under light microscope,significal interstitial infiltration of inflammatory cells was found in alveoli and interstitial tissue of the lung,and destruction of alveolar tissue,alveolar wall thinning,and even rupture to fuse into bullae,and bleeding into alveoli in different degress could be observed. Compared with the normal control group,forced vital capacity(FVC),forced expiratory volume in 0.3 second(FEV0.3),FEV0.3/FVC,peak expiratory flow(PEF)in model group were significantly decreased〔FVC(mL):8.04±2.03 vs. 9.97±2.14,FEV0.3(mL):6.16±2.23 vs. 8.84±2.12,FEV0.3/FVC:0.70±0.09 vs. 0.85±0.11,PEF(mL/s):33.56±4.76 vs. 40.14±5.64, P<0.05 or P<0.01〕. Serum IL-6 was obviously increased(ng/L:93.17±20.96 vs. 76.28±13.24,P<0.05), IL-10 was significantly decreased(ng/L:78.62±15.17 vs. 104.34±19.46,P<0.01),and CD4+CD25+FoxP3+Treg was significantly diminished〔(2.75±0.83)% vs.(4.16±1.14)%,P<0.01〕in model group compared with those in the normal control group. The expression of Foxp3 protein in lung tissue in model group was significantly down-regulated compared with that in the normal control group(gray scale:0.38±0.15 vs. 0.63±0.11,P<0.01), and RORγt and IL-17 protein expressions were significantly up-regulated〔RORγt(gray scale):0.96±0.23 vs. 0.47±0.11,IL-17(gray scale):1.02±0.24 vs. 0.34±0.08,both P<0.01〕. Correlation analysis showed that FEV0.3 was positively correlated with Foxp3(r=0.585,P<0.05),and FEV0.3/FVC was negatively correlated with IL-6 and RORγt(r=-0.655,r=-0.607,both P<0.05). PEF was positively correlated with Treg(r=0.573, P<0.05),and negatively correlated with IL-17(r=-0.198,P<0.05). IL-6 was negatively correlated with Foxp3(r=-0.603,P<0.05),and positively correlated with RORγt(r=0.588,P<0.05). IL-10 was positively correlated with Treg(r=0.573,P<0.05). Treg was positively correlated with Foxp3(r=0.607,P<0.05), and negatively correlated with IL-17(r=-0.569,P<0.05). Foxp3 was negatively correlated with RORγt(r=-0.591, P<0.05). RORγt was positively correlated with IL-17(r=0.578,P<0.05). Conclusion There is a relationship among decreased pulmonary function,inflammation and imbalance of Foxp3/Treg and RORγt/Th17 in COPD.
ABSTRACT
Objective To observe the effects of Liuwei Buqi capsules on CD4+CD25+forkhead box protein 3+(Foxp3+)regulatory T cells(Tregs),helper T cells(Ths)in patients with chronic obstructive pulmonary disease (COPD). Methods Eighty COPD patients admitted into the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from January 2012 to May 2013 were randomly divided into a traditional Chinese medicine (TCM)experimental group(40 cases)and a western medicine control group(40 cases);there were 40 research volunteers enrolled as healthy controls. After admission,both the TCM experimental group and western medicine control group were given conventional western medicine comprehensive treatment. Additionally,in the TCM experimental group,Liuwei Buqi capsules were given orally once 3 pills(0.4 g/pill)and twice a day,while in the western medicine control group nifedipine was given,once 10 mg and 3 times a day,30 days constituting one therapeutic course for both groups. BODE evaluation means that the body mass index(B),the degree of airflow obstruction(O),the scores of dyspnea(D)and excercise capacity(E)of COPD patients were evaluated. A spirometer was used to observe the changes in the patient's pulmonary function to detect the dyspnea score. The γ-interferon(IFN-γ),interleukin (IL-4 and IL-17)in serum were detected by enzyme-linked immunosorbent assay(ELISA). The expression of Treg of peripheral blood was detected by flow cytometry. Results Compared with healthy control group,lung function parameters,IL-4,CD4+CD25+Treg and CD4+CD25+Foxp3+Treg of peripheral blood were increased,while dyspnea score,BODE score,the expressions of IFN-γ,Th1/Th2,IL-17 in serum were decreased significantly in the COPD group(P<0.05 or P<0.01). After treatment,compared with those before treatment in the two groups, the forced expiratory volume in 1 second(FEV1),FEV1/forced vital capacity(FVC),IL-4,CD4+CD25+Treg, CD4+CD25+Foxp3+ Treg were all increased,while dyspnea score,BODE score,IFN-γ,Th1/Th2,IL-17 were all decreased(P<0.05 or P<0.01),and the peak expiratory flow(PEF)was decreased in western medicine control group and increased in TCM experimental group. After treatment,the comparisons of ratio of FEV1/FVC〔(78.12±14.96)%vs.(67.52±10.39)%〕,BODE score(1.07±0.72 vs. 1.77±0.74),IL-17(μg/L:40.80±8.97 vs. 48.22±6.51) Th1/Th2(1.05±0.23 vs. 1.42±0.21)and CD4+CD25+Foxp3+ Treg〔(6.61±2.26)% vs.(5.25±2.03)%〕between the two groups were all of statistical significant difference(all P<0.05). Conclusion Liuwei Buqi capsules can improve COPD symptoms by up-regulating the expressions of CD4+CD25+Foxp3+ Treg,IL-4,and down-regulating the expressions of IFN-γ,IL-17 to correct the balance of Th1/Th2 in patients with COPD.