Effect of Maxing Shigantang on Airway Injury and EGFR Expression in Asthma Model Rats / 中国实验方剂学杂志

ObjectiveTo explore the optimal formula of Maxing Shigantang in regulating epidermal growth factor receptor（EGFR）expression and alleviating airway injury in asthmatic rats and to reveal the underlying mechanism. MethodSD male rats were randomly divided into normal group, model group, dexamethasone group (5×10-4 g·kg-1) and Maxing Shigantang 1∶0.5, 1∶1, 1∶2 groups (group A, B, C, 10 g·kg-1), with 8 rats in each group. The other groups except the normal group received nebulization of 2% acetylcholine chloride and 0.4% histamine phosphate for the modeling of asthma. One hour before modeling， the normal group and the model group were given the same amount of normal saline， and the other groups were given the same amount of corresponding drugs， once a day for 7 days. On the 7th day， the model was established and the incubation period of asthma was recorded. The rats were then immediately anesthetized， and arterial blood and tracheal tissue were collected. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the levels of interleukin-2 （IL-2）， interleukin-4 （IL-4）， and tumor necrosis factor-α （TNF-α） in serum. Pathological sections were prepared for the observation of the pathological changes of tracheal tissues and the ultrastructure of epithelial cells in each group. Terminal-deoxynucleotidyl transferase-mediated nick-end labeling （TUNEL） was adopted to detect epithelial cell apoptosis， and in situ hybridization and Western blot were employed to determine the mRNA and protein levels of epidermal growth factor receptor （EGFR）， respectively. ResultCompared with the model group， groups A， B and C prolonged the incubation period of asthma （P<0.05，P<0.01）. Compared with the control group， the model group showed declined IL-2 level （P<0.01）， risen IL-4 and TNF-α levels （P<0.05，P<0.01）， increased airway pathology score， collagen volume fraction， and airway epithelial cell apoptosis index （P<0.01）， and up-regulated mRNA and protein levels of EGFR in trachea tissue （P<0.01）. Compared with the model group， group A showed increased IL-2 level （P<0.05） and declined IL-4 （P<0.05，P<0.01） level， and group B showed declined IL-4 level （P<0.05）. The level of TNF-α in groups A， B， and C declined compared with that in the model group （P<0.01）. Maxing Shigantang repaired the tracheal tissue to different degrees （P<0.05）. Among the three groups， group A inhibited tracheal fibrosis （P<0.05） and had the most significant effect of repairing the ultrastructural changes of airway epithelial cells. Groups A， B and C all inhibited the apoptosis of airway epithelial cells （P<0.05）. All the three groups inhibited the up-regulation of EGFR mRNA level （P<0.05，P<0.01）， and groups B and C inhibited the up-regulation of EGFR protein level （P<0.05，P<0.01）. ConclusionMaxing Shigantang can inhibit the abnormal changes of airway epithelial structure， alleviate airway injury， and can down-regulate the expression of EGFR in the tracheal tissue of asthma model rats. In this study， the optimal compatibility of Maxing Shigantang to repair airway epithelial injury in asthmatic rats was group A， with the Ephedrae Herba-Armeniacae Semen Amarum-Glycyrrhizae Radix et Rhizoma-Gypsum Fibrosum ratio of 1∶0.5∶4∶1.

Acupuncture combined with western medication for ocular myasthenia gravis: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the clinical efficacy between acupuncture combined with western medication and simple western medication for ocular myasthenia gravis (OMG), and to explore its possible mechanism.@*METHODS@#A total of 60 patients of ocular myasthenia gravis were randomized into an acupuncture combined with western medication group (30 cases, 1 case dropped off) and a western medication group (30 cases, 2 cases dropped off). Oral pyridostigmine bromide tablet and prednisone acetate tablet were given in the western medication group. On the basis of the treatment in the western medication group, Tongdu Tiaoqi acupuncture (acupuncture for unblocking the governor vessel and regulating qi ) was applied at Baihui (GV 20), Fengfu (GV 16), Hegu (LI 4), Zusanli (ST 36), etc. in the acupuncture combined with western medication group, once a day, 6 days a week. The treatment was given 8 weeks in both groups. Before and after treatment, the OMG clinical absolute score was observed, electrophysiological indexes of orbicularis oculi (value of mean jitter, percentage of jitter >55 μs and percentage of blocks) were measured by single-fiber electromyography (SFEMG), serum levels of acetylcholine receptor antibody (AChR-Ab), interferon-gamma (IFN-γ) and interleukin-4 (IL-4) were detected by ELISA method.@*RESULTS@#After treatment, the OMG clinical absolute scores, values of mean jitter, percentages of jitter >55 μs, percentages of blocks and serum levels of AChR-Ab, IFN-γ and IL-4 were decreased compared before treatment in both groups (P<0.05), and those in the acupuncture combined with western medication group were lower than the western medication group (P<0.05).@*CONCLUSION@#Acupuncture combined with western medication can effectively improve ptosis, palpebra superior fatigability, eye movement disorder and neuromuscular junction dysfunction in patients with ocular myasthenia gravis, the therapeutic effect is superior to simple western medication. Its mechanism may be related to down-regulating serum levels of AChR-Ab, IFN-γ and IL-4 and promoting the recovery of orbicularis oculi function.

Mechanism of Total Saponins from Dioscoreae Nipponicae Rhizoma to Treat Gouty Arthritis by Regulating M1/M2 Polarization of Macrophages / 中国实验方剂学杂志

Objective:To illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis. Method:A total of 72 male SD rats were randomly divided into four groups： normal group， model group， total saponin group （160 mg·kg^-1）， celecoxib group （43.3 mg·kg^-1）， with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin（HE） staining. Immunohistochemistry method was used to detect the protein expression change of CD68， interleukin-4（IL-4）， inducible nitric oxide synthase （iNOS） and transforming growth factor-<italic>β</italic>₁（TGF-<italic>β</italic>₁）. Result:HE staining results showed that the inflammation of the model group was most obvious on the third day after modeling， and the disease was in the acute stage. On day 5， the inflammation was alleviated， and on day 8， the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue， and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd，5th and 8th day of administration （<italic>P</italic><0.01）. Compared with the model group， the total saponins group could reduce the expression of CD68 and iNOS （<italic>P</italic><0.05，<italic>P</italic><0.01）on the 3rd day of administration， and significantly reduced them expression on the 5th and 8th days （<italic>P</italic><0.01）. Compared with the normal group， IL-4 and TGF-<italic>β</italic>₁ expression were increased in the model group when the drug was given for three days（<italic>P</italic><0.01）. Total saponin group could enhance IL-4 expression（<italic>P</italic><0.05）and decreased the TGF-<italic>β</italic>₁ expression（<italic>P</italic><0.01）. Compared with normal group， the expression of IL-4 in the model group decreased on the 5th and 8th day of administration （<italic>P</italic><0.01）， and the expression of TGF-<italic>β</italic>₁ in the model group decreased on the 5th day of administration（<italic>P</italic><0.01）. Compared with the model group， the total saponins group could increase the expression of IL-4 and TGF-<italic>β</italic>₁ at 5 d and 8 d after administration （<italic>P</italic><0.01）. Conclusion:Total saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.

Effect of electroacupuncture combined with pill on sex hormone and Th2 cytokines in patients of decreased ovarian reserve function with liver-kidney deficiency / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) combined with pill on clinical symptoms, levels of serum sex hormone and Th2 cytokines in patients of decreased ovarian reserve function (DOR) with liver-kidney deficiency, and to compare the efficacy between EA combined with pill and pill alone.@*METHODS@#Sixty patients with DOR were randomly divided into an observation group (30 cases, 2 cases dropped off) and a control group (30 cases, 1 case dropped off). The patients in the control group were treated with pill, 1 pill each time, 3 times a day. Based on the treatment of the control group, the patients in the observation group were additionally treated with acupuncture at Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Zigong (EX-CA 1), Zusanli (ST 36), Sanyinjiao (SP 6), Taixi (KI 3) and Taichong (LR 3); EA was applied at bilateral Zusanli (ST 36) and Sanyinjiao (SP 6), with continuous wave, in frequency of 20 Hz and current intensity of 1 to 4 mA, for 20 min. The treatment was given 3 times a week. All the patients terminated treatment during menstrual period, and the treatment was given for 3 continuous menstrual cycles. The menstrual condition score and systemic symptom score were compared between the two groups before and after treatment. The levels of serum sex hormones on 2nd to 3rd day of menstruation, including follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2), and the serums levels of interleukin (IL) -4 and IL-10 secreted by Th2 cytokines were compared between the two groups before and after treatment.@*RESULTS@#After the treatment, the menstruation condition scores and systemic symptom scores in the two groups were reduced (<0.05), and the scores in the observation group were lower than those in the control group (<0.05). After the treatment, the levels of serum FSH, LH and FSH/LH were reduced (<0.05), and the E2 levels were increased in the two groups (<0.05), and the levels of FSH, LH in the observation group were lower than those in the control group (<0.05), and the E2 level was higher than that in the control group (<0.05). After the treatment, the levels of serum IL-4 and IL-10 in the two groups were increased (<0.05), and the levels of IL-4 and IL-10 in the observation group were higher than those in the control group (<0.05).@*CONCLUSION@#EA combined with pill could significantly improve menstruation, systemic symptoms and serum sex hormone levels in patients of decreased ovarian reserve function with liver-kidney deficiency, which may restore ovarian function by up-regulating the expression of Th2 cytokines.

Effect of Bletillae Rhizoma Polysaccharide on PI3K/Akt in Gastric Tissue of GU Model Rats / 中国实验方剂学杂志

Objective::To investigate the effect of Bletillae Rhizoma polysaccharide on the expressions of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) gene protein and its mediated cytokines interleukin-2 receptor (IL-2R) and interleukin-4 (IL-4) in gastric tissue of rats with gastric ulcer (GU). Method::Sixty SPF Wistar rats were randomly divided into blank group and model group.The GU model was replicated by direct acetic acid cauterization in model group.The GU model rats were randomly divided into five groups: model group, positive control group, and large, medium and small-dose Bletillae Rhizoma polysaccharide groups, with 10 rats in each group.Rats in blank group and GU model group were given 10 mL·kg-1·d-1 distilled water by gavage, rats in large, medium and small-dose groups were given 0.5, 0.25, 0.125 g·kg-1·d-1 Bletillae Rhizoma polysaccharide by gavage, while rats in positive control group were given 0.3 g·kg-1·d-1 ranitidine by gavage for 15 days.Serum nitric oxide (NO) content, pepsinase activity and cytokines IL-2R and IL-4 levels in rats of each group were measured by enzyme-linked immunosorbent assay (ELISA), PI3K and Akt mRNA expressions were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), and PI3K and Akt protein expressions were detected by Western blot. Result::Compared with the blank group, the contents and gene expressions of cytokines IL-2R and IL-4 in gastric tissue were significantly increased, and the PI3K and Akt genes and protein expressions were significantly increased, with statistical significance (P<0.01). Compared with GU model group, the content and gene expressions of IL-2R and IL-4 in large, medium and small-dose Bletillae Rhizoma polysaccharide groups were decreased significantly, and the PI3K and Akt gene and protein expressions were decreased significantly in large-dose Bletillae Rhizoma polysaccharide group, while those in large and medium-dose Bletillae Rhizoma polysaccharide groups were decreased significantly (P<0.05, P<0.01). Conclusion::Bletillae Rhizoma polysaccharide can protect gastric mucosa by down-regulating PI3K and Akt gene and protein expressions and inhibiting abnormal secretion of cytokines IL-2R and IL-4.

Clinical Efficacy of Modified Xiaofengsan and Its Effect on Cytokines in Persistent Asthma Patients with Wind-asthma Pattern / 中国实验方剂学杂志

Objective: To observe the clinical efficacy of modified Xiaofengsan and its effect on cytokines in persistent asthma patients with wind-asthma pattern. Method: The 120 eligible patients with chronic and persistent asthma were randomly divided into control group (60 cases) and treatment group (60 cases). Patients in control group received the budesonide/formoterol (160 μg/4.5 μg, 2 inhales/time, bid). Patients in treatment group additionally received the modified Xiaofengsan combined with budesonide/formoterol (160 μg/4.5 μg). The treatment course was 1 month in both groups. The clinical efficacy of the two groups was observed. Before and 3 months after treatment, asthma control test (ACT) and the average times of acute exacerbation in 6 months before and after treatment between two groups were observed; percentage of forced expiratory volume in first second and its predicted value (FEV1%), variety ratio of Peak expiratory flow (PEF), eosinophilic granulocyte (EOS) count in peripheral blood and the levels of interleukin-4(IL-4), interleukin-6(IL-6) and interleukin-13(IL-13) in serum were measured before and after treatment respectively. In addition, the safety of the two groups was evaluated. Result: The total effective rate of the treatment group was higher than that of the control group (P1% in treatment group were higher(PPConclusion: Modified Xiaofengsan combined with budesonide/formoterol could relieve symptoms of asthma, improve pulmonary function and lower the levels of IL-4, IL-6 and IL-13 in serum.

Effect of moxibustion combined with autohemotherapy of acupoint injection for life quality of allergic rhinitis withdeficiency of lung and spleen:a randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of moxibustion combined with autohemotherapy of acupoint injection and simple moxibustion for allergic rhinitis(AR) withdeficiency of lung and spleen and to explore its mechanism.</p><p><b>METHODS</b>By random number table,75 patients who matched the criteria were assigned into a moxibustion combined with autohemotherapy of acupoint injection group(a combination group,37 cases) and a moxibustion group(38 cases). In the combination group,moxibustion was used at bilateral Yingxiang(LI 20) and injection bila-teral Feishu(BL 13),Pishu(BL 20) and Zusanli(ST 36). In the moxibustion group,moxibustion was applied bilaterally at all the above acupoints. Acupoint injection was given twice a week,and moxibustion once a day,seven days as a course. The patients were observed for four courses and followed up half a year after treatment. Rhinoconjunctivitis quality of life questionnaire(RQLQ) was compared before and after treatment and at follow-up. The short-term and long-term effects as well as the changes of interferon-(IFN-),interleukin-4(IL-4) and immunoglobulin E(IgE) were observed in the two group.</p><p><b>RESULTS</b>After treatment,the obviously effective rate of the combination group was 75.68%(28/37),which was higher than 52.63%(20/38) of the moxibustion group(<0.05). But at follow-up, the obviously effective rate of the moxibustion group was 47.37%(18/38),which was better than 24.32%(9/37) of the combination group(<0.05). Life quality of the two group in all aspects after treatment and at follow-up were apparently improved compared with the baseline(<0.01,<0.05). After treatment,the life quality,sleep condition,non nasal-eye symptoms,behavior issue of the combination group were superior to those of the moxibustion group(all<0.05),and at follow-up,the above outcomes,the eye and nasal symptoms as well as emotional response of the mo-xibustion group were better(all<0.05). There was statistical significance in the two group about serum IFN-,IgE,IL-4 before and after treatment(<0.01,<0.05),and significance existed between the two groups(<0.05).</p><p><b>CONCLUSIONS</b>Moxibustion combined with autohemotherapy of acupoint injection can effectively improve the short-term quality of life of AR withdeficiency of lung and spleen. The mechanism may be related to balancing the immune response of Th1 and Th2.</p>

Gene Therapy of Inner Ear Injection with Bone -Marrow Mesenchymal Stem Cells Decorated with Interleukin-4 Gene on Immune-Mediated Inner Ear Disease in Guinea Pigs / 听力学及言语疾病杂志

Objective To evaluate the gene therapeutic effects of guinea pigs model with immune -mediated inner ear disease(IMIED)after locally injection of bone -marrow mesenchymal stem cells(BMSCs) decorated by in-terleukin-4 gene .Methods Guinea pigs were immunized with keyhole limpet hemocyanin (KLH) and caused 55 animal models ,divided into five groups ,each group with 11 animals :groupA(BMSCs carrier) ,group B(BMSCs emp-ty -carrier control group) ,group C(recombinent lentivirus IL -4 gene) ,group D(lentivirus empty -carrier control group) ,group E(simulation operation control group) .Groups were all injected with the corresponding suspension (20 μl)[includs BMSCs ceas of (1 .5~2 .0) × 106 ,the concentration of (entivirus) is 0 .5 × 108 pfu] by the scala tympani window into the inner ear .The fluorography immunohistochemistry test and enzyme immunohistochemistry test for the situation of IL -4 gene express and productive protein distribution in inner ear .Auditory functions and the KLH level of guinea pigs blood were monitored respectively by auditory brainstem responses (ABRs) and ELISA test .Results The threshold of ABR wave Ⅲ decreased in group A ,group B and group C .The result were more significant in group A and group B than that in group C ,but results in group A was more prominent (P<0 .05) . The results of immunohistochemistry test showed that fluorescence positive BMSCs mainly scattered in scala tymani and scala vestibule .The microscope results showed that for the group A ,B and C ,there were only few foccule and red and white blood cells in scala tympani floc ,but for group D and group E ,with different levels of labyrinthine hy-drops and some mononuclear cells around the spiral ganglion and small blood vessels .Conclusion Restructuring lentiviral vector with IL -4 gene can be successfully transfected into BMSCs in vitro ,compared to inplangting into inner ear in scala tympani approach ,the cells can migrate and generate gene product of IL -4 ,to significantly im-prove the auditory functions and inflammatory reaction of inner ear disease ,and BMSCs can be used as a carrier to migrate to the damaged part with therapeutic gene .

Effects of Mycobacterium Phlei on Funcitonal States of Th_1/Th_2 in Patients with COPD and Preventive Effects on AECOPD / 药物流行病学杂志

Objective:To investigate the effects of Mycobacterium Phlei on functional states of Th_1/Th_2 in patients with chronic obstructive pulmonary disease(COPD),and clinical effects on the prevention of acute exacerbation of chronic obstructive pulmonary diseases(AECOPD).Method:60 patients with COPD of gradeⅡwere randomly divided into a treatment group and controlled group.The controlled group were given a routine therapy,while the treatment group were given utilins in addition to the routine treatment.The contents of?-interferon(IFN-?) and interleukin-4(IL-4) in their peripheral blood and their clinical efficacy were observed before and after the treatment.Result:Before and after the treatment, IFN-?/IL-4 ratio in the treatment group was 1.39?0.20 and 1.94?0.31 respectively(P

Study of the sex differences in experimental autoimmune encephalomyelitis / 中国康复理论与实践

@# ObjectiveTo explore the mechanism causing the sex differences in experimental autoimmune encephalomyelitis (EAE).MethodsEAE model was established with PLP 139-151 in SJL/J mice. The sex differences of illness state were evaluated by neurologic score, and that of response of peripheral lymphocytes to autologous antigens were detected by methods of MTT and ELISA.ResultsThe onset time of female SJL mice was 15±2.1 days less than that of male ones (22±4.3 days), and the feature of diseases course of female mice was relapse after recovery, but that of male mice was transient. No significant differences were observed in T cell responses and interferon-γ productions between male and female mice (P>0.05). Male mice secreted more interleukin-4 than female mice (P<0.01).ConclusionThe sex differences of EAE in mice are due to peripheral lymphocytes secreting more interleukin-4 in male mice.

The Effect of Ginkgo on Interleukin4(IL-4) and Interleukin5(IL-5) in Serum of Asthmatic mice / 中国医师杂志

Objective To observe the effect of Ginkgo on IL-4,IL-5 in serum of asthmatic mice. Methods 40 mouse were randomly divided into three groups(control group, sensitization group, and therapeutic group). The mouse asthmatic models were established by sensitizing with aerosolized ovalbumin(OVA). IL-4,IL-5 in serum in 16 sensitization mouse and 8 control mouse were measured by ELISA, respectively. We injected 0.15ml Ginkgo solution in the abdominal cavity of 16 therapeutic mouse before the episode of asthma. After that, they were challenged with aerosolized OVA and IL-4,IL-5 in serum were measured by ELISA. Results The levels of IL-4,IL-5 in serum in control group were (12.965?2.450)pg/ml, (13.398?4.460)pg/ml, respectively, (16.889?4.835)pg/ml, (19.471?7.207)pg/ml in sensitization, (13.695?4.068)pg/ml, (14.774?5.749)pg/ml in therapeutic group. In the sensitization group, the levels of IL-4,IL-5 in serum were higher than those of the control group( P

An immunohistochemical study on the time-dependent expression of IL-10 and IL-4 during skin incised wound healing in mice / 中国法医学杂志

Objective To explore the temporal expressions of interleukin-10(IL-10)and interleukin-4(IL-4)and their relationship with the wound age during skin incised wound healing in mice.Methods Immunohistochemical and image-analysis techniques were employed in vital skin wounds 0.5h～168h after incision and postmortem incision 0.5h～6h after death.Results Weak expression of IL-10 was detected at 0.5h after incision,which increased subsequently,and peaked at 3h post-injury.Then decreased to the pre-incision level at 24h,but peaked again at 72h after injury.Immunostaining revealed that epidermal cells and infiltrating mononuclear cells were the major source of IL-10.IL-4-positive cells were observed in the dermis at 24h,and reached the maximum level at 96h,which maintained until 168h post-injury.IL-4 was mainly expressed in the fibroblast-like cells of the granulation tissue and dermis.In addition,faint IL-10-positive cells were merely identified in the epidermal cells in the postmortem incision skin within 1～3h after incision.No IL-4-postitive cells were found in the postmortem incision skin.Conclusion The time-dependent expressions of IL-10 and IL-4 during cutaneous wound healing may be used for the estimation of wound age.

The studies on the properties of dendritic cells from human cord blood and malignant pleural effusions / 肿瘤研究与临床

Objective To investigate the culture and proliferation of dendritic cells from human cord blood and malignant pleural effusions and identify the phenotype and the function of them in vitro. Methods Existing DCs were obtained from cord blood and malignant pleural effusions. The DCs were cultured with IL-4, GM-CSF and TNF-?. The morphological properties of DCs were observed by invert optical microscope. The phenotypes of DCs were analysed by flow cytometry. The proliferative bioactivities of T cells activated by DCs were observed with mixed lymphocyte reaction (MLR). Results No remarkable differences were found in the morphology and cellular phenotype of the DC, derived from two different sources, but there was remarkable difference between their culture time. The DCs from two different sources could activate homologous lymphocytes to proliferate mostly. But the capacity to stimulate the proliferation of homologous lymphocytes had remarkable difference. Conclusion Mature DCs could be induced from cord blood and malignant pleural effusions.