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ObjectiveTo investigate the therapeutic effects and difference in the effects of Arisaematis Rhizoma （AR） before and after processing （i.e.， Arisaematis Rhizoma Preparatum， ARP） with Zingiberis Rhizoma Recens-Alumen on allergic asthma in rats and to provide a basis for the theory of processing improving the efficacy. MethodA rat model of allergic asthma was established in 70 SD rats by intraperitoneal injection of ovalbumin （OVA）-aluminum hydroxide. The rats were administrated with the aqueous extracts of AR （1.2， 0.3 g∙kg-1） and ARP （1.2， 0.3 g∙kg-1） aqueous extracts by gavage， and montelukast sodium （0.001 g∙kg-1） was used as the positive drug. The T helper cell type 1/type 2 （Th1/Th2） ratio in the serum and bronchoalveolar lavage fluid （BALF） and percentages of inflammatory cells in BALF were determined. Polymerase chain reaction （PCR） was employed to determine the mRNA level of mucin 5AC （MUC5AC） in the lung tissue. The pathological changes in the lung tissue were observed by hematoxylin-eosin （HE） staining and PAS staining. Immunohistochemical assay was employed to measure the expression of c-Jun amino-terminal kinase （JNK）， extracellular signal regulated protein kinase （ERK）， and p38 mitogen-activated protein kinase （p38 MAPK） in rat lung tissue. Western blot was employed to determine the protein levels of ERK， p-ERK， JNK， p-JNK， p38， p-p38 in the lung tissue. The effects of AR and ARP were compared based on overall desirability. ResultCompared with the blank group， the levels of interleukin-12 （IL-12） and γ interferon （IFN-γ） in serum and BALF of rats in the model group were significantly lower （P<0.05， P<0.01）， and the levels of interleukin-4 （IL-4）， interleukin-5 （IL-5） and interleukin-13 （IL-13） were significantly higher （P<0.05， P<0.01）. Compared with the model group， the serum and BALF contents of IL-12 and IFN-γ in rats in the montelukast sodium group， high-dose AR group and high-dose ARP group were significantly higher （P<0.05， P<0.01）， and the contents of IL-4， IL-5 and IL-13 were significantly lower （P<0.05， P<0.01）， and the serum contents of IFN-γ in rats in the low-dose AR group and low-dose ARP group were in BALF was significantly higher （P<0.05） and IL-4 and IL-13 were significantly lower （P<0.05， P<0.01）， the percentages of macrophages， lymphocytes， neutrophils， and eosinophils were reduced in BALF， and the expression of JNK/ERK/p38 MAPK signaling pathway and MUC5AC protein was inhibited in lung tissues. Overall assessment of the normalized analysis revealed that the ARP group was slightly more potent than the AR group after administration of the same dose. ConclusionAR and ARP can effectively treat allergic asthma by inhibiting JNK/ERK/p38 MAPK signaling pathway， and the effect is better after concoction， which can provide data support for its "concoction efficiency".
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Objective: To evaluate the effect of short-Term and long-Term immunization of recombinant disorganized muscle protein-1 (rDIM-1) in rodents against human filarial parasite Brugia malayi. Methods: Recombinant Brugia malayi DIM-1 (rDIM-1bm) protein was cloned, expressed and purified using a Ni-NTA affinity column. Mastomys coucha were immunized with rDIM-1bm in three immunization schedules: short-Term (3-dose of rDIM-1bm), and long-Term (booster doses till 3-And 6-week) and subsequently challenged with infective third-stage larvae of filarial parasite Brugia malayi (L3). Microfilaraemia was monitored in L3 exposed groups on day 90 post larval inoculation (p.l.i.) and continued till day 205 p.l.i. On day 205 p.l.i. all the infected animals were killed and total worm burden was estimated. Cellular proliferative response, macrophage activity, nitric oxide (NO) release, specific IgG and its subtypes, IgE, IgA and Th1 (IFN-γ, TNF-α and IL-2) and Th2 (IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release were determined. Results: Of the 3 different immunization schedules, short-Term immunization (3-dose schedule) showed better reduction in microfilarial burden (36%-63%) in the peripheral circulation, adult worm load (52%), whereas long-Term immunization (3-And 6-week schedule) exerted less effect on peripheral microfilariae count (9%-58%), and adult worm burden (9%-12.5%). Short-Term immunization resulted in upregulation of cellular proliferation, macrophages activity, NO release, specific IgG, IgG1, IgG2a, IgG2b, IgE and IgA levels and both Th1 (IFN-γ, TNF-α and IL-2) and Th2 (IL-4, IL-5, IL-6, IL-10 and IL-13) cytokine release whereas long-Term immunization (3-And 6-week schedule) exerted less effect on parasite burden and showed mixed immunological responses. None of the rDIM-1bm administration schedules induced any pathology in lymphoid tissues, or alteration in mast cell number and granularity. Conclusions: The short-Term immunization with rDIM-1bm (3-dose schedule) induces robust immune responses and protects the host from filarial parasite infection.
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Objective: To investigate the effect of modified Renseng Yangrong Tang combined with chemotherapy on serum tumor markers and Th1/Th2 type cytokines expression in cervical cancer patients. Method: A total of 114 patients with cervical cancer admitted in our hospital from December 2014 to December 2017 were selected as the study subjects. According to the random number table method, the patients were divided into two groups. The control group (57 cases) was treated with conventional chemotherapy. The treatment group (57 cases) was treated with modified Renseng Yangrong Tang in addition to the conventional chemotherapy. Traditional Chinese medicine (TCM) symptom scores, serum tumor markers[carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCCA), carbohydrate antigen 125 (CA125)]and Th1/Th2 type cytokine[interferon gamma-interferon (IFN-γ), interleukin-4 (IL-4), interleukin-2 (IL-2), interleukin-6 (IL-6) levels], clinical efficacy after treatment 3 months, and adverse reaction rate of two groups were observed. Result: There was no significant difference in TCM symptom scores, serum tumor markers and Th1/Th2 cytokines between the two groups before treatment. After treatment, the TCM symptom scores, CEA, SCCA, CA125, IL-6, and IL-4 levels in the treatment group were lower than those in control group and before treatment (Pγ were higher than those in the control group and before treatment (PPχ2=4.524, 4.150, PConclusion: Modified Renseng Yangrong Tang can significantly relieve clinical symptoms, reduce the level of serum tumor markers, improve cellular immunity, improve the recent therapeutic effect, and reduce adverse reactions in patients with cervical cancer chemotherapy.
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Objective: To discuss the efficacy of Jiawei Changfengyin with retention enema therapy for acute radiation proctitis and investigate its influence on regulating levels of helper T cell(Th)1/Th2. Method: One hundred and twenty-eight patients were randomly divided into control group and observation group by random number table. Patients in control group (64 cases) got montmorillonite powder (3.0 g), dexamethasone (10 mg) and normal saline (100 mL), with retention enema therapy. Patients in observation group (64 cases) got Jiawei Changfengyin(Changfengyin+Xileisan+pearl powder), with retention enema therapy. Both groups of patients received enema once every night, and the treatment course was 4 weeks in both groups. Before and after treatment, scores of main symptoms, proctoscopy, routine examination of stool+occult blood, and KPS scores of quality of life were graded. Levels of Th1 cell factors[interleukin(IL)-1β, IL-8 and interferon-γ(IFN-γ)], Th2 cell factors (IL-4 and IL-10) were detected. Result: The total effective rate for comprehensive clinical efficacy was 95.31% in observation group, higher than 82.81% in control group (χ2=5.132, PPZ=2.764, PPPβ, IL-8 and IFN-γ in observation group were lower than those in control group, while levels of IL-4 and IL-10 were higher than those in control group (PConclusion: Jiawei Changfengyin can relieve symptoms of acute radiation proctitis, promote healing of rectal mucosa, improve quality of life, and regulate Th1/Th2 cytokines, with good repairing effect for intestinal mucosa.
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Objective:To elucidate the relationship between HLA-DQB1 allele polymorphism as well as the expression level of Th1/Th2 cytokines with familial clustering of hepatocellular carcinoma ( HCC) to provide some evidence for the seeking susceptibility gene or resistant gene of HCC in Guangxi yao,China.Methods:With the same sexuality,age ±5 year,40 members whose families have had two or more HCC patients( high-occurrence families) were selected as the case group,and 40 members whose families have no any cancer patient were selected as the controls.Peripheral blood samples were collected to extract DNA,PCR-SSP was used to detect HLA-DQB1 alleles and ELISA was used to detect IL-2,IL-4 and IL-10.Results:(1) The gene frequency of the HLA-DQB1*02/09 alleles in the case group was higher than that in the controls(P0.05 ).( 2 ) The gene frequency of alleles HLA-DQB1 in HBsAg positive group and HBsAg negative group were never significant difference (P>0.05).(3)The expression levels of IL-4,IL-10 in the case group was higher than that in the control ( P<0.05 ).( 4 ) The expression level of IL-10 in the positive group of the HLA-DQB1*02 allele was higher than that in the negative group of the HLA-DQB1*02 allele ( P<0.05 ).( 5 ) The expression level of IL-4 in the positive group of the HLA-DQB1*09 allele was higher than that in the negative group of the HLA-DQB1*09 allele( P<0.05) .Con-clusion:(1) HLA-DQB1*02/09 seem to be susceptibility genes of hepatocellular carcinoma in high HCC incidence areas of Guangxi yao.(2) There may be not significant correlation bewteen HLA-DQB1 alleles and the susceptibility of HBV infection in high HCC incidence areas of Guangxi yao.( 3 ) The imbalance of IL-4, IL-10 might be associated with familial clustering of hepatocellular carcinoma in Guangxi yao.(4)The imbalance of IL-10 might be due to the carrying of HLA-DQB1*02;the imbalance of IL-4 might be due to the carrying of HLA-DQB1*09.Through the same approaches,these might lead to the phenomenon of familial aggregation of HCC in Guangxiyao.
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Objective To investigate the changes of human leukocyte antigen-G (HLA-G) protein expression and Th1/Th2 type cytokines in intrahepatic cholestasis of pregnancy (ICP) and their relativity to the etiology of ICP. Methods Peripheral blood and placental tissues were obtained from 26 ICP patients (the ICP group) and 22 normal pregnant women (the NP group) in the operation room for Cesarean birth. Immunohistochemistry was used to detect the expression of HLA-G protein in the placental tissues. Meanwhile we tested the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4) by enzyme-linked immunosorbent assay (ELISA) in the peripheral blood and checked the levels of TBA in the serum.Results TBA level in the ICP group was (27.05±6.08) μmol/L, significant higher than that in the NP group (4.35±2.68)μmol/L (P＜0.01). The positive expression of HLA-G protein in extravillous trophoblast in the ICP group was significantly lower than that in the NP group (P＜0.01). The mean optical density (MOD) of positive expression of HLA-G protein in the placenta tissues in the ICP group (52.91±7.19) was significantly lower than that in the NP group (69.26±7.72) (P＜0.01). The concentration of TNF-α was significantly higher in the ICP group (101.31±19.30) pg/mL than that in the NP group (54.51±23.72) pg/mL (P＜0.01). The concentration of IL-4 was lower in the ICP group (22.16±6.55) pg/mL than that in the NP group (31.69±8.25) pg/mL (P＜0.01). The ratio of TNF-α/IL-4 was higher in the ICP group (4.52±1.91) than that in the NP group (1.72±0.61) (P＜0.01). There was a negative correlation between the MOD of HLA-G protein and TNF-α (r=-0.98, P＜0.01) in the ICP group. No correlation with IL-4 and TNF-α/IL-4 was seen (P＞0.05). There was a positive correlation between TBA and TNF-α (r=0.99, P＜0.01), and a negative correlation between TBA and the MOD of HLA-G protein (r=-1.00, P＜0.01) in the ICP group. No correlation with IL-4 and TNF-α/IL-4 was seen (P＞0.05). Conclusion There is an imbalance of Th1/Th2 cytokines to the Th1 type in the peripheral blood of ICP patients. The expression of HLA-G protein in the placenta of ICP patients decreases, leading to an increase of Th1 type cytokines that may be one of the reasons for liver destroy in ICP.
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Objective To study the effects of purified rabies vaccine for human use (RV) on specific Th1/Th2 cytokines in human. Methods Twenty cases were injected intramuscularly with 5 full doses of RV. PBMCs were isolated from the blood sample collected at day 0, 14, 45 after the RV inoculation. Neutralizing antibody was determined by ELISA, and the proliferation of lymphocyte by in vitro test. The levels of RV specific IFN-γ, TNF, IL-2,IL-4, IL-5, IL-10 in the culture supernatants were detected by cytometric bead array (CBA). Results The neutralizing antibody was tested positive in 19 cases 45 days after inoculation and 1 case after 60 days, with the positive rate reaching 100%. After stimulation with RV, the lymphocyte transformation index at day 14, 45 in cases were significantly higher than those day of 0 (P< 0.05), and similar results were confirmed with IFN-γ, IL-2, IL-4, IL-5 tested by CBA (P<0.05). Condusion The RV could induce humoral and antigen-specific cellular immune responses in human, tested by showing good protective effect on rabies virus.
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Objective Cervical heterotopic heart transplantation model was established in different inbred strains of mice with modified cuff technique. Inbred male Balb/c mice and C57BL/6 mice were selected as donors and recipients, respectively. Mice were randomly assigned into four groups: control group (the donor hearts were perfused through coronary artery with 200 μl, 0℃～4℃ St. Thomas Ⅱ solution during 2 to 3 min, then they were immersed in it for 15 min), CsA group ( the donor hearts were perfused with the same method as for the control's and intraperitoneal injection of CsA 5 mg· g-1 · d -1 was given after surgery ), H2-B1 transfection group (the donor hearts were perfused through coronary artery with 200 μl, 0℃ -4℃ St. Thomas Ⅱ solution contained with 30 μg H2-Bl plasmid vectors during 2 to 3 min, then they were immersed in it for 15 min ), and H2-B1 + CsA group ( the donor hearts were perfused with St. Thomas Ⅱ solution contained H2-Bl gene plasmid and intraperitoneal injection of CsA was given after surgery as mentioned above. ). At 1,3 and 7 days after transplantation, three allografts were harvested at each time points in all of the groups, respectively, for pathological examination and analysis of CD40 expression with immunohistochemistry assays. The expression of Th1/Th2 cytokines were also determined with flow cytometry. The survival time of rest allografts were observed. Results Histological features for rejection were observed more apparent in the grafts of control group than those in other groups, especially those in H2-Bl + CsA group. The expression of CD40 in H2-Bl + CsA group and CsA group was lower significantly than that of the control group ( P ＜0.01 ), so was the expression of CD40 in the H2-Bl group as compare with that of the control group (P ＜0.05). No significant difference between H2-Bl group and CsA group (P ＞0.05 ) at 7 days was observed. The expression of IL-2, TNF-α (Th1 cytokines) in control group was much higher than that in other groups, and the expression of IL-4 ( Th2 cytokine) in control group was much lower ( P ＜0.05 ). The level of IL-4 in CsA group increased significantly at 3 days ( P ＜ 0.05 ), with a peak level at 7 days after transplantation (P＜0.01). The survival time of grafts was significantly prolonged in CsA group (P＜0.01), H2-Bl group (P＜0.05) and H2-Bl+CsA group(P＜0.01). Conclusion Treating the donor hearts with H2-Bl plasmid vectors at the time of transplantation may suppress rejection in the heart allografts and prolong the survival time through some presumed mechanisms such as preventing upregulation of CD40 expression, relucing the production of IL-2 and TNF-α, increasing the production of IL-4, and as a result, inducing immune tolerance, as well as improving the function of transplanted heart grafts.
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Objective To analyze the effect of cyclosporin A(CsA), rapamycin(RPM) and macophenolic acid(MPA) on the co-stimulated lymphocytes, CD28 and CD40, and their production of Th1/Th2 cytokine, IL-2, IFN-γ, IL-4, IL-10 and IL-12. Methods The experimental groups were divided into ①mono-stimulating and co-stimulating groups: CD3 mAb mono-stimulating (group a),CD3 mAb+CD28 mAb co-stimulating (group b), CD3 mAb+CD28 mAb+CD40 L mAb co-stimulating(group c), CD3 mAb+CD28 mAb+CTLA4 mAb co-stimulating (group d). ②CsA groups: 300 ng/ml of CsA was added to group a, b, c and d. ③RPM groups: 300 ng/ml of RPM was added to group a,b, c and d. ④MPA groups:300 ng/ml of MPA was added to group a, b, c and d. The cytokine production was measured by ELISA.Results The co-stimulated CD28 and CD40 Th1/Th2 cytokines production of IFN-γ, IL-2, IL-4 and IL-10 were significantly increased. Compared with group a, IFN-γ increased from (248.91±11.20)ng/ml to (555.08±24.42)ng/ml and (548.19±33.06)ng/ml, IL-2 increased from (29.48±8.61)ng/ml to (1100.82±99.29)ng/ml and (842.23±29.31)ng/ml, IL-4 increased from (32.29±6.76)ng/ml to (116.02±15.03)ng/ml and (147.28±18.07)ng/ml, IL-10 increased from (147.01±10.47)ng/ml to (291.79±12.47)ng/ml and (302.52±35.18)ng/ml,respectively, P＜0. 01. Compared group b with group c, the Th1 cytokines production was decreased.IL-2 and IL-12 decreased (P＜0.05). The Th2 cytokine IL-4 production was increased (P＜0. 05).CTLA4 mAb and three other immunosuppressants, CsA, RPM and MPA, inhibited co-stimulated lymphocyte's both cytokines Th1 and Th2 production. The inhibitory effect of CsA on Th1/Th2 cytokine production was more significant than RPM and MPA did. The co-inhibitory effect of CTLA4 mAb and CsA was observed as well. The increased co-stimulated CD28 and CD40 IL-12 production could be suppressed by MPA. CsA and RPM had no inhibitory effect on the IL-12 production.Conclusions CD28/CD40 co-stimulatory pathway plays the key role in lymphocyte activation and Th1/Th2 cytokine production. CsA, RPM and MPA can inhibit co-stimulated lymphocyte's Th1 and Th2 cytokine production. CsA and CTLA4 mAb have co-inhibitory effect on co-stimulated lymphocyte's Th1/Th2 cytokines production. CD40 L mAb decreases the Th1 cytokines production(including IL-12) and increases the Th2 (mainly IL-4) production, which may be the mechanism of its longevity effect on allograft.
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Objective To explore the effect of gaseous signal molecule hydrogen sulfide(H2S)on the expression of lung Muc5ac and Th1/Th2 cytokines in ovalbumin(OVA)-induced asthma rats and explore the effect of H2S on asthma.Methods Twenty-six healthy SD rats were randomly divided into 3 groups:control group(Group A,n=8),asthma group(Group B,OVA induced,n=9)and NaHS group(Group C,OVA induced rats treated with NaHS,n=9).Twenty-four hours after treatment,rats were anatomized to measure serum interleukin(IL)-4,interferon-? and the levels of infiltration into inflammatory cells around bronchus were observed,which were scored with the optical microscope.The expression of lung Muc5ac was determined by immunohistochemical staining.Results The score of the levels of infiltration into inflammatory cells around bronchus expressed by median was 1 score in group A,4 score in group B and 2 score in group C.There were dramatically statistics significance among the 3 groups(H=13.75 P
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Objective:To study the immunologic and endocrinologic mechanism of Chinese herbs in the treatment of abortion Methods:SD rats injected with bromocriptine 0 125 mg/d subcutaneously(sc)during day 6~8 of pregnancy were used as aborting model Uteruses of the rats treated with or without Chinese herbs were taken out at day 12 of pregnancy for counting embryoes and determining mRNA levels of Th1/Th2 type cytokines in decidua by semiquantity RT PCR Results:The insufficiency of PRL and P in serum during gestation was related to embryo miscarriage remarkably A predominant Th1 response at the maternal fetal interface is harmful to pregnancy Aborting rate induced by bromocriptine was decreased after treating with Chinese herbs Chinese herbs can increase serum PRL and P , then trigger a predominant Th2 response in the model Conclusion:With it's effect of increasing PRL and P in serum and the Th2 bias response at the maternal fetal interface, Chinese herbs can reverse the abortion effect of bromocriptine
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107 copies/mi group. Conclusion: The presence of high amount of HBVDNA in serum is associated with suppressed co-stimulatory and regulatory effect of IL-12 to Th1/Th2 profilation