Effect of Buzhong Yiqitang on Th17/Treg Immune Imbalance and Notch1 Signaling Pathway in AIT Mice / 中国实验方剂学杂志

ObjectiveTo explore the effect of Buzhong Yiqitang on the immune imbalance of helper T cell 17 （Th17）/regulatory T cell （Treg） and Notch1 signaling pathway in mice with autoimmune thyroiditis （AIT）. MethodA total of 60 8-week-old NOD.H-2h4 mice were randomly divided into the normal group， model group， western medicine group （selenium yeast tablet， 32.5 mg·kg-1）， and low-dose （4.78 g·kg-1·d-1）， middle-dose （9.56 g·kg-1·d-1）， and high-dose （19 g·kg-1·d-1） Buzhong Yiqitang groups， with 10 mice in each group. The normal group was fed with distilled water， and the other groups were fed with water containing 0.05% sodium iodide for eight weeks. After the animal model of AIT was formed spontaneously， the mice were killed under anesthesia after intragastric administration for eight weeks. Serum anti-thyroglobulin antibodies （TGAb）， thyroid-stimulating hormone （TSH）， free triiodothyronine （FT3）， and free thyroid hormone （FT4） were detected by enzyme-linked immunosorbent assay （ELISA）， and thyroid tissue changes were observed by hematoxylin-eosin （HE） staining. The mRNA and protein expressions of retinoid-related orphan receptor-γt （RORγt）， interleukin （IL）-17， forkhead box P3 （FoxP3）， IL-10， Notch1， and hair division-related enhancer 1 （Hes1） in thyroid tissue were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the normal group， the thyroid structure of the model group was severely damaged， and lymphocytes were infiltrated obviously. The levels of serum TGAb， FT3， and FT4 contents were significantly increased， and TSH content was significantly decreased （P<0.01）. The mRNA and protein expression levels of RORγt， IL-17， Notch1， and Hes1 were significantly increased， while those of FoxP3 and IL10 were significantly decreased in the model group （P<0.01）. Compared with the model group， thyroid structural damage and lymphocyte infiltration were improved in the treatment groups， and serum TGAb， FT3， and FT4 contents were significantly decreased. TSH content was increased， and mRNA and protein expression levels of RORγt， IL-17， Notch1， and Hes1 were decreased. mRNA and protein expression levels of FoxP3 and IL-10 were increased to different degrees （P<0.05， P<0.01）， and the middle-dose Buzhong Yiqitang group had the most significant intervention effect. ConclusionBuzhong Yiqitang can alleviate the thyroid structural damage in AIT mice， and its mechanism may be related to improving the abnormal differentiation of Th17/Treg immune cells and inhibiting the activation of the Notch1 signaling pathway.

Mechanism of catalpol regulating Th17 cell differentiation by interfering PKM2/LDHA expression / 中国药房

OBJECTIVE To investigate the mechanism of catalpol affecting the differentiation of helper T cell 17 （Th17） by interfering the expressions of pyruvate kinase M2 （PKM2） and lactate dehydrogenase A （LDHA）. METHODS The naive CD4+ T cells were selected from the spleen of C57BL/6 mice， and were differentiated into Th17 cells by adding directional differentiation stimulants for 72 hours. At the same time， the cells were treated with 0 （directed control）， 20， 40 and 80 μg/mL catalpol. The flow cytometry was used to detect the proportion of Th17 cell differentiation in cells； the colorimetric method was adopted to detect the levels of pyruvate and lactate in cell culture supernatant； mRNA expressions of retinoid-related orphan nuclear receptor gamma t （RORγt）， PKM2 and LDHA were detected by qRT-PCR method； Western blot was used to detect the expression levels of PKM2， LDHA， signal transducer and activator of transcription 3 （STAT3）， and phosphorylated STAT3 （p-STAT3） proteins in cells. RESULTS Compared with the directed control group， after 72 hours of treatment with 20， 40， 80 μg/mL catalpol， the differentiation ratio of Th17 cells were decreased by 6.74%， 8.41%， 9.24%， and the levels of pyruvate and lactate in the cell culture supernatant， the mRNA expressions of PKM2， LDHA and RORγt as well as the protein expressions of PKM2 and LDHA and the phosphorylation of STAT3 were significantly reduced （P＜0.05）. CONCLUSIONS Catalpol can reduce the glycolysis level by down-regulating the expressions of PKM2 and LDHA， thereby inhibiting the differentiation of Th17 cells.

Adalimumab combined with Dexamethasone intravitreal implant in the treatment of refractory noninfectious uveitis macular edema / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of adalimumab combined with dexamethasone intravitreal implant in the treatment of refractory non-infectious uveitis macular edema(UME).METHODS: A total of 92 cases(131 eyes)of refractory non-infectious UME patients admitted to our hospital from January 2020 to January 2022 were selected and randomly divided into control group, with 46 cases(63 eyes)treated with dexamethasone intravitreal implant and observation group, with 46 cases(68 eyes)treated with adalimumab subcutaneous injection combined with dexamethasone intravitreal implant. The best corrected visual acuity(BCVA), central retinal thickness(CRT), vitreous opacity and Th17/Treg cytokines were measured before and after treatment, and the occurrence of adverse reactions was recorded.RESULTS: Totally 3 cases(4 eyes)were lost to follow-up. After treatment for 1, 3, 6 and 12 mo, BCVA was improved in both groups compared with that before treatment, and CRT, vitreous opacity score, serum interleukin(IL)-17 and IL-22 levels were decreased compared with those before treatment, and serum transforming growth factor-β(TGF-β)and IL-10 levels were increased compared with those before treatment. BCVA in the observation group was better than that in the control group, and CRT, vitreous opacity score, serum IL-17 and IL-22 levels were lower than those in the control group, and serum TGF-β and IL-10 levels were higher than those in the control group(all P&#x0026;#x003C;0.05). During treatment and follow-up, no serious adverse reactions occurred in both groups.CONCLUSION: Adalimumab combined with dexamethasone intravitreal implants in the treatment of refractory non-infectious UME can significantly subside the macular edema, reduce vitreous opacity and improve visual acuity.

Intervention research status of traditional Chinese medicine monomers on autoimmune diseases by regulating Th17/Treg axis based on gut microbiota / 中国药房

Autoimmune diseases（ADs） are diseases in which the body’s immune tolerance is impaired， causing damage to its tissues. The pro-inflammatory helper T cell 17 （Th17） and anti-inflammatory regulatory T cell （Treg） are functionally antagonistic to each other， and the immune imbalance between them and the imbalance of related inflammatory factors are closely related to the occurrence of a variety of ADs. Plenty of evidence has shown that gut microbiota can regulate Th17/Treg differentiation， rebuild immune tolerance and delay the ADs process through regulating cytokine production， transcription factor expression and energy metabolism. This paper reviews the intervention effects of traditional Chinese medicine（TCM） monomers on the common ADs by regulating Th17/Treg differentiation balance based on intestinal flora： ulcerative colitis，rheumatoid arthritis and diabetes mellitus type 1. It is found that its mechanism of action may be to restore the balance of pro-inflammatory factors and anti-inflammatory factors to alleviate intestinal mucosal barrier damage， reduce synovial angiogenesis and improve pancreatic β cell destruction， which provides some ideas for the prevention and treatment of ADs with integrated traditional Chinese and western medicine.

Research status of traditional Chinese medicine regulating Th17/Treg balance in the treatment of rheumatoid arthritis / 中国药房

Rheumatoid arthritis （RA） is a refractory autoimmune disease that can cause symmetrical polyarticular disease. The key mechanism of its occurrence and development is the dysequilibrium of helper T cell 17 （Th17）/regulatory T cell （Treg） balance. Therefore， reconstructing Th17/Treg balance may be a new strategy for the treatment of RA. Traditional Chinese medicine has significant advantages in the treatment of RA such as integrity， multi-target， multi-link and multi-path. This paper summarizes the basic and clinical studies on the regulation of Th17/Treg balance in the treatment of RA by traditional Chinese medicine in the past five years， and finds that the active components/sites of traditional Chinese medicine such as flavonoids， alkaloids and terpenes have unique advantages in the regulation of Th17/Treg balance. The traditional Chinese medicine compound formula interferes with Th17/Treg balance by exerting the effects of dispelling wind， dehumidifying， removing blood stasis， unblocking collaterals， relieving pain， dispersing cold and strengthening health. The effect of external treatment of traditional Chinese medicine is obvious and can be used as a clinical adjuvant therapy for RA； related mechanisms of action include regulating the production of inflammatory factors， regulating the expression of transcription factors and interfering with the activation of signaling pathways. However， the existing research has the shortcomings of insufficient mechanism research， few clinical research， limited external treatment research of traditional Chinese medicine， and lack of combination therapy research， which need to be improved by follow- up research.

Mechanism of Bushen Huatan Prescription in Regulating Th17/Treg Balance in Ovariectomized Osteoporosis Rats / 中国实验方剂学杂志

Objective:To study the effect of Bushen Huatan prescription on helper T cell 17 （Th17）/T regulatory cells （Treg） balance of immune T cell subsets in the prevention and treatment of postmenopausal osteoporosis. Method:Sixty 6-month-old female SD rats were randomly divided into sham operation group， model group， estradiol valerate group （0.184 mg·kg^-1） and Bushen Huatan prescription low， medium and high groups （4.7， 9.4， 18.8 g·kg^-1） according to the random number table. All the groups except the sham operation group received ovariectomy to make postmenopausal osteoporosis model. Intragastric administration was started 1 week after operation， and the rats in model group and sham operation group received equal volume of normal saline， once a day for 12 weeks. Microcomputed tomography （Micro CT） was then used to detect bone mass and microstructure of rats， the contents of Forkhead box protein （Foxp3） and retinoic acid related nuclear orphan receptor （ROR<italic>γ</italic>t） in serum were detected by enzyme-linked immunosorbent assay （ELISA）， the mRNA expression levels of Foxp3 and ROR<italic>γ</italic>t in bone tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot was used to detect the protein expression of Foxp3 and ROR<italic>γ</italic>t in bone tissues， the number of Th17 and Treg cells in each group was analyzed and compared by flow cytometry. Result:Compared with the sham operation group， the bone mass and trabeculae of the model group decreased （<italic>P</italic><0.01）， the bone microstructure was destroyed， the concentration of Foxp3 in serum decreased， the concentration of ROR<italic>γ</italic>t increased （<italic>P</italic><0.01）， the mRNA and protein expression levels of Foxp3 in bone tissues decreased， ROR<italic>γ</italic>t increased， the number of Treg cells in bone tissues decreased， number of Th17 cells increased （<italic>P</italic><0.01）， and Th17/Treg ratio increased （<italic>P</italic><0.01） in model group. Compared with the model group， the bone mass in each treatment group increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， Foxp3 concentration in serum increased， ROR<italic>γ</italic>t concentration decreased （<italic>P</italic><0.01）， the mRNA and protein expression levels of Foxp3 in bone tissues increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， but no statistical difference was shown in mRNA expression between low dose group and the model group. In addition， the mRNA and protein expression of ROR<italic>γ</italic>t decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， number of Treg cells increased， number of Th17 cells decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， and Th17/Treg ratio decreased in treatment groups （<italic>P</italic><0.01）. Conclusion:Bushen Huatan prescription can increase bone mass， improve bone microstructure， increase the number of Treg cells and decrease the number of Th17 cells in ovariectomized rats. It is concluded that Bushen Huatan prescription may play a role in preventing and treating postmenopausal osteoporosis by regulating Th17/Treg balance.

Clinical Efficacy of Sheti Zhiqiu Decoction Combined with Intradermal Needling in Treatment of Allergic Rhinitis with Deficient Cold of Lung Qi Syndrome / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of Sheti Zhiqiu decoction combined with intradermal needling in treatment of allergic rhinitis （AR） of deficient cold of lung Qi syndrome and its effect on cytokines of helper T cell 17 （Th17） and regulatory T cells （Treg）. Method:A total of 105 patients with AR of deficient cold of lung Qi syndrome were randomly divided into traditional Chinese medicine （TCM） group， combination group and western medicine group， with 35 cases in each group. The TCM group was treated with Sheti Zhiqiu decoction， 1 dose a day. The combination group was treated with intradermal needling in addition to the therapy of the TCM group. The selected acupoints were Yintang， bilateral Yingxiang， Fengchi， Feishu and Zusanli. The needles were retained for 3 days before being replaced. The western medicine group was treated with mometasone furoate nasal spray， 100 μg/time at each side of the nasal cavity， <italic>qd</italic>， and desloratadine citrate tablet， 8.8 mg/time， <italic>qd</italic>. Three groups were treated for 4 weeks and followed up for 3 months. Nasal and ocular symptom scores and rhinoconjunctivitis quality of life questionnaire （RQLQ） scores of patients in three groups were observed before and after treatment. Levels of serum interleukin-17 （IL-17）， interleukin-10 （IL-10） and transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁） were measured before and after treatment. The clinical efficacy and safety were evaluated， and the disease recurrence rate was observed during the follow-up period. Result:Compared with before treatment， nasal and ocular symptom scores and total score， and RQLQ scores and total score of patients in three groups were significantly decreased after treatment （<italic>P</italic><0.01）. Compared with the TCM group and the western medicine group after treatment， except for eyes itching/foreign body sensation/red eyes score， nasal and ocular symptom scores and total score， and RQLQ scores and total score of the combination group were lower（<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with before treatment， level of serum IL-17 of patients in three groups was significantly decreased （<italic>P</italic><0.01）， while levels of IL-10 and TGF-<italic>β</italic>₁ were significantly increased after treatment （<italic>P</italic><0.01）. Compared with the TCM group and the western medicine group after treatment， level of serum IL-17 was lower， whereas levels of IL-10 and TGF-<italic>β</italic>₁ were higher in the combination group （<italic>P</italic><0.05，<italic>P</italic><0.01）. The clinical efficacy of the combination group was better than that of the TCM medicine group and the western medicine group （Z=-2.207，Z=-2.185，<italic>P</italic><0.05）. There was no significant difference in recurrence rate between combination group and the TCM group， and the recurrence rate of both groups was lower than that of the western medicine group（<italic>χ</italic>²=5.020，<italic>χ</italic>²=4.835，<italic>P</italic><0.05）. There was no significant adverse reaction during the treatment period in three groups. Conclusion:Sheti Zhiqiu decoction combined with intradermal needling is effective in treatment of patients with AR of deficient cold of lung Qi syndrome. It can significantly relieve patients' symptoms， improve patients' quality of life and reduce disease recurrence. It may play a role by regulating immune balance of Th17/Treg of patients and improving their immune function.

Traditional Chinese Medicine Intervention on Th17/Treg Balance in Chronic Obstructive Pulmonary Disease： A Review / 中国实验方剂学杂志

Chronic obstructive pulmonary disease （COPD）， a common clinical chronic respiratory disease， has a long course and is intractable. It is closely related to many factors， such as immune imbalance. Helper T cell 17 （Th17）， an immune-promoting cell， and regulatory T cell （Treg）， an immunosuppressive cell， maintain the balance of the immune microenvironment together. In the course of COPD， the proportion of Th17 cells usually increases， while the proportion of Tregs that inhibit Th17 activity decreases. Their coordination and balance are critical in the inflammatory and immune processes of COPD. At present， COPD is mainly treated with nasal inhalation preparations and oral drugs by western medicine. In spite of a certain therapeutic effect， side effects of drugs and heavy economic burden are becoming increasingly prominent. Modern research shows that traditional Chinese medicine （TCM） has the characteristics of few side effects， stable curative effect， and multi-target regulation， and it is advantageous and promising in the prevention and treatment of COPD. In recent years， a large number of TCM clinical and experimental trials on the intervention of Th17/Treg balance in COPD have been launched. Substantial pieces of evidence confirm that the intervention of Th17/Treg balance is an important potential target of TCM in the treatment of COPD. This study reviewed the previous research on the intervention effect of single Chinese medicine， effective components of Chinese medicine， and Chinese medicinal compound on Th17/Treg balance in COPD to comprehensively reveal the potential target of Th17/Treg balance in COPD for clinicians and scientific researchers， promote relevant research， and provide references for the rational application of TCM in the prevention and treatment of COPD.

Effect of medicated thread moxibustion of Chinese Zhuang Minority medicine on colonic inflammatory impairment and Th17/IL-17F signaling in rats with ulcerative colitis / 针刺研究

OBJECTIVE: To observe the effect of medicated thread moxibustion of Zhuang Minority medicine on helper T cell 17 (Th17)/ Interleukin-17F(IL-17F) signaling pathway in ulcerative colitis (UC) rats, so as to explore its mechanisms underlying improvement of UC. METHODS: Forty male SD rats were randomly divided into normal control, model, medication and medicated thread moxibustion (MTM) groups, with 10 rats in each group. The UC model was induced by free drinking of 4% dextran sulfate sodium (DSS) for 10 d. After successful modeling, rats of the medication group were treated by gavage of salazosulfapyridine (SASP). Medicated thread moxibustion was applied to unilateral "Tianshu" (ST25) and "Qihai" (CV6) alternatively for rats of the MTM group, once daily for 14 d. The body mass, stool shape, and fecal occult test were recorded and conducted daily to perform disease activity index (DAI) score. H．E. staining was used to display pathological changes of the colon tissue. The Th17 cells and IL-17F and retinoic acid related orphan receptor γ t (ROR-γt) in the colon tissue were detected by flow cytometry, and enzyme-linked immunosorbent assay (ELISA), respectively, and the expression levels of RORγt and IL-17F mRNA in colon tissue were detected by quantitive real-time PCR. RESULTS: After modeling, the DAI score, colonic Th17 percentage, RORγt and IL-17F contents, and RORγt and IL-17F mRNA expression were significantly increased in the model group in contrast to the normal control group (P0.05, except RORγt and IL-17F mRNA expression). H．E. staining showed disappearance of goblet cells, infiltration of a large number of inflammatory cells, exfoliation of the epithelial tissue and edema of colonic mucosal in rats of the model group, which was relatively milder in both medication and MTM groups. CONCLUSION: Medicated thread moxibustion of Zhuang Minority medicine can reduce the inflammatory damage of colon tissue in UC rats, which is associated with its effects in suppressing the expression of RORγt, production of Th17 cells, and secretion of pro-inflammatory factor IL-17F in colon tissue.

Ginkgolide A reduces airway inflammation in a neutrophil-predominant murine model of asthma via suppression of immune response of helper T cell 17 / 中国中西医结合急救杂志

Objective To determine the effects of ginkgolide A (GA) in a neutrophil-predominant murine model of asthma and explore underlying mechanisms. Methods Thirty-five female BALB/c mice were randomly divided into sham operation group (Sham group), asthma group, dexamethasone intervention control group (DEX group), low dose GA intervention group (L-GA group) and high dose GA intervention group (H-GA group), with 7 mice in each group. The asthma model was induced by intraperitoneal injection of 20 μg ovalbumin (OVA) and 75 μL Fluorine complete adjuvant (FCA) on day 0, 14 and 21, and challenged 30 minutes with 5% OVA atomization on days 22-24 consecutively; phosphate buffer (PBS) was sensitized and stimulated in Sham group. The mice in L-GA group and H-GA group were intraperitoneally injected with GA of 40 mg/kg and 80 mg/kg at 1 hour before each challenge, while the mice in DEX group were intraperitoneally injected with dexamethasone of 1 mg/kg. After 24 hours of the last OVA stimulation, the airway resistance was measured at the time of 0, 3, 6, 12, 25, 50 g/L acetylmethacholine aerosol stimulation. The total number of cells and cell classification in bronchoalveolar lavage fluid (BALF) were counted. The transforming growth factor-β1 (TGF-β1) and interleukin-17 (IL-17) in BALF were detected by enzyme linked immunosorbent assay (ELISA). The proportion of helper T cell 17 (Th17) to CD4+ T cell in lung tissue was detected by flow cytometry, and the pathological characteristics of lung tissue were evaluated. Results Compared with the Sham group, the airway hyper responsiveness (AHR), the total cells, the neutrophil counts, the levels of TGF-β1, IL-17 in BALF, and the proportion of Th17 cells in the lung tissue in the asthma group were significantly increased, obvious inflammatory cell infiltration and collagen fiber deposition around airway were observed, and airway inflammation score and mucus score were significantly increased. Compared with the asthma group, low and high doses of GA significantly reduced AHR, and there was a significant difference in airway resistance at the time of 50 g/L acetylmethacholine stimulation (cmH2O·s-1·mL-1: 5.29±0.40, 3.99±0.57 vs. 7.34±0.77, both P < 0.05); the total cells, neutrophil counts, and levels of TGF-β1, IL-17 in BALF, and the proportion of Th17 cells in lung tissue were significantly decreased [total cells count (×104/L): 21.00±1.00, 17.00±1.02 vs. 27.50±2.50; neutrophil count (×104/L): 12.600±0.600, 10.610±0.210 vs. 16.875±1.125; TGF-β1 (ng/L): 371.40±107.80, 289.60±70.76 vs. 551.90±68.34; IL-17 (ng/L): 60.75±11.79, 44.77±7.09 vs. 122.50±38.87; the proportion of Th17 cells: (5.53±0.40)%, (3.76±1.10)% vs. (8.30±1.19)%, all P < 0.05]; inflammatory cell infiltration around the airway and mucus secretion was significantly reduced, airway inflammation score and mucus score were significantly decreased (2.16±0.28, 1.16±0.28 vs. 3.77±0.25; 1.33±0.58, 1.17±0.29 vs. 3.67±0.58, all P < 0.05). The AHR, total cells, neutrophil counts, and IL-17 level in BALF, the proportion of Th17 cells in lung tissue and airway inflammation score decreased more obviously with the increase of GA dosage (all P < 0.05). For index mentioned above, no significant differences were observed between DEX group and asthma group. Conclusion GA treatment was effective in a murine model of neutrophil-predominant asthma via inhibiting response in the immune cells Th17.

Preventive effect of octreotide combined with netilmicin sulfate on postoperative infection in suppurative appendicitis and its effect on Th17/Treg imbalance / 中国生化药物杂志

Objective To investigate the preventive effect of octreotide combined with netilmicin sulfate on postoperative infection in suppurative appendicitis and its effect on helper T cell 17(Th17)/regulatory T cells(Treg)imbalance.Methods 148 patients with acute suppurative appendicitis in Xiangshan Hospital of Traditional Chinese Medicine from August 2014 to May 2016 were randomly divided into observation group(n=74)and control group(n=74).All were underwent laparoscopic appendectomy surgery,the control group took netilmicin sulfate combined with ornidazole for anti-infection after operative,while the observation group added octreotide acetate subcutaneously.The levels of serum interleukin(IL)-4,IL-6,IL-10,IL-17,IL-23,transforming growth factor β(TGF-β),interferon γ(IFN-γ)and Th17,Treg cell ratio and expression levels of related transcription factorbefore and after treatment were detected,Th17/Treg change were analyzed.Results The levels of IL-4,IL-6,IL-10,IL-17,TGF-β were decreased,IFN-γ were increased after treatment(P<0.01),the improvement of those indicators in the observation group were supior than control group(P<0.05).Th17,Treg cell ratio and expression levels of related transcription factor orphan nuclear receptorγt(RORγt),Forkhead box protein P3(Foxp3)were decrease,Th17/Treg was reduced after treatment(P<0.01),those indexes in observation group were lower than those in control group(P<0.05).The wound infection,intra-abdominal infections and other adverse events had no significant difference in two groups.Conclusions Octreotide combined with netilmicin sulfate can prevent postoperative infection in suppurative appendicitis,regulate Th17/Treg imbalance,restore immune function,and no significant adverse reactions.

Role of Foxp3/Treg and RORγt/Th17 cell imbalance in rat model of chronic obstructive pulmonary disease / 中华危重病急救医学

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.

Detection of Th17 cells in peripheral blood of patients with multiple myeloma and its clinical significance / 肿瘤

Objective: To examine the Th17 cells (helper T cells 17) in peripheral blood of patients with MM (multiple myeloma) and explore its clinical significance. Methods: The peripheral blood samples from 39 patients with MM and 30 healthy volunteers were collected. The proportion of Th17 cells in CD4 + T cells was determined by flow cytometry. The expression of ROR-γt (retinoid-related orphan receptor-γt) mRNA, a specific transcription factor of Th17 cells, was detected by RT-PCR. The plasma concentration of IL-17 (interleukin-17) was measured by ELISA assay. Results: The proportion of Th17 cells, the expression level of ROR-γt mRNA and the concentration of IL-17 in peripheral blood in patients with MM were higher than those in the healthy volunteers [(3.509±0.594)% vs (2.913±0.846)%, 0.568±0.177 vs 0.468±0.163, 27.200±4.666 vs 23.758±7.410 pg/mL; all P values 0.05), but correlated with ISS (international staging system) stage, β2-microglobulin level and the therapeutic response (P < 0.05). Conclusion: Th17 cells may be involved in the occurrence and development of MM. Detection of proportion of Th17 cells may contribute to the ajudgement of disease severity, therapeutic response and the prognosis of patients with MM. Copyright © 2013 by TUMOR.