Effect of acupuncture on acupoint "Yingxiang-Hegu" on Th1, Th2 cytokines and T-bet/GATA-3 of allergic rhinitis rats

Objective:To explore the effect of"Yingxiang-Hegu"on Th1,Th2-related cytokines and[2]transcription factors T-bet and GATA-3 in rats with allergic rhinitis.Methods:Rats were randomly divided into three groups:blank group,model group and acupoint group.The rat model of ovalbumin(OVA)AR was established,and the general condition of the rats was observed and scored.Acupuncture intervention was performed on the acupoint group on the second day after successful modeling,once per day for 20 min for 10 d.After intervention,the general behavior,behavioral score and histomorphological changes of nasal mucosa were observed.Eosinophils(EOS)were counted under microscope after nasal lavage smear staining,and the contents of total IgE,IFN-γ,IL-12,IL-4 and IL-5 in serum were detected by ELISA.Westernblot and IHC were used to detect the protein level and positive protein expression of specific transcription factors T-bet and GATA-3 in rat nasal mucosa.Results:After the establishment of the model,except for the blank group,the behavioral observation scores of rats in the model group and acupoint group were more than 5 points,indicating that the model was successful.After acupuncture intervention on acupoint"Yingxiang-Hegu",the behavioral score of rats in the acupoint group and western medicine group was significantly lower than that in the model group(P<0.05).Microscopic examination showed that the structure of nasal mucosa in the model group was obviously damaged,cilia were arranged discontinuously,uneven,local congestion and swelling,a large number of epithelial cells exfoliated and necrotic,goblet cell proliferation,obvious inflammatory cell infiltration.The pathological degree of nasal mucosa in the pair point group was significantly less than that in the model group.Compared with the model group,the levels of IFN-γ,IL-2 and IL-12 in serum were significantly increased,while IgE,IL-4,IL-5 and IL-6 were significantly decreased,GATA-3 protein and positive expression in nasal mucosa were significantly decreased and T-bet was significantly increased after acupuncture.Conclusion:Acupuncture at"Yingxiang-Hegu"can effectively improve the nasal sensitive symptoms and control nasal inflammation in AR rats.The mechanism may be that acupuncture at Yingxiang-Hegu can up-regulate the expression of T-bet,decrease the level of GATA-3,promote the production of Th1 cytokines and inhibit the synthesis and secretion of Th2 cells,thus restoring the immune balance of Th1 and Th2.

Expression status of GATA3 and mismatch repair proteins in upper tract urothelial carcinoma

GATA binding protein 3 (GATA3) and mismatch repair (MMR) deficiency contribute to the development of urothelial carcinoma. However, the combined expression of GATA3 and microsatellite instability (MSI) in upper tract urothelial carcinoma (UTUC) and its prognostic value have not been investigated. Here, we immunohistochemically stained GATA3 and MMR proteins in 108 UTUC samples. GATA3 was positive in 74 cases, and its expression was significantly lower than in adjacent benign urothelium (P<0.001). Loss of GATA3 expression was statistically associated with adverse clinicopathologic parameters, such as advanced stage, lymphovascular invasion, neural invasion, lymph node metastasis, and extensive necrosis. Cancer-specific survival (CSS, P=0.028) and disease-free survival (DFS, P=0.024) were significantly shorter in patients with GATA3 negative tumors than in patients with GATA3 positive tumors. The absence of MMR proteins was observed in 8.3% of the cases, and focal staining was identified in 13.0%. When using “lax criteria” which resulted in counting cases as negative where MMR staining was in fact focally positive (<5%), we found that GATA3 was inversely associated with MSI (P=0.005). Moreover, GATA3−/microsatellite stability (MS) tumors were correlated with advanced pT stage (P<0.001) and poor outcome (P=0.019 for CSS, P=0.016 for DFS) compared with GATA3+/MSI ones. The GATA3−/MSI cases had unfavorable clinical outcomes compared with GATA3+/MSI cases (P=0.008 for CSS, P=0.023 for DFS). This finding raises a question as to whether GATA3 interacts with MSI through the TGF-β signaling pathway and regulates UTUC progression.

Clinical Auditory Phenotypes Associated with GATA3 Gene Mutations in Familial Hypoparathyroidism-deafness-renal Dysplasia Syndrome

Background： Hypoparathyroidism-deafness-renal dysplasia （HDR） syndrome is an autosomal dominant disorder primarily caused by haploinsufficiency of GATA binding protein 3 （GATA3） gene mutations, and hearing loss is the most frequent phenotypic feature. This study aimed at identifying the causative gene mutation for a three-generation Chinese t;amily with HDR syndrome and analyzing auditory phenotypes in all familial HDR syndrome cases. Methods： Three affected family members underwent otologic examinations, biochemistry tests, and other clinical evaluations. Targeted genes capture combining next-generation sequencing was pertbrmed within the family. Sanger sequencing was used to confirm the causative mutation. The auditory phenotypes of all reported familial H DR syndrome cases analyzed were provided. Results： In Chinese family 712 l, a heterozygous nonsense mutation c.826C〉T （p.R276＊） was identified in GA TA3. All the three affected members suffered from sensorineural deafness and hypocalcemia; however, renal dysplasia only appeared in the youngest patient. Furthermore, an overview of thirty HDR syndrome families with corresponding GATA3 mutations revealed that hearing impairment occurred earlier in the younger generation in at least nine familial cases （30%） and two thirds of them were found to carry premature stop mutations. Conclusions： This study highlights the phenotypic heterogeneity of HDR and points to a possible genetic anticipation in patients with HDR, which needs to be further investigated.

Prognostic signifi cance of transcription factors FOXA1 and GATA-3 in ductal carcinoma in situ in terms of recurrence and estrogen receptor status

Aim:The aim was to analyze the expression of novel biological transcription markers,forkhead-box A1(FOXA1),GATA binding protein 3(GATA-3),and established markers such as Ki-67(MIB-1)and human epidermal growth factor receptor 2(HER2)in estrogen receptor(ER(+))and ER(-)ductal carcinoma in situ(DCIS)patients with/without recurrence.Methods:Two hundred and ninety-one cases of DCIS were retrieved from our pathology database,with complete data available for 219 cases.The follow-up period is from 1988 to 2009.Recurrence is defined in terms of DCIS or invasive carcinoma(IC).No recurrence was seen in 88%(196/219)of cases;12%(26/219)had a recurrence(IC:13,DCIS:13).We are reporting the results of biological marker expression in terms of recurrence and ER status.Results:Our study demonstrates strong expression of GATA-3 in the ER(+)DCIS in recurrence and nonrecurrence groups similar to previously described in IC.A reduced expression of GATA-3 was observed in ER(-)recurrence and nonrecurrence groups.A strong HER2 protein expression,as well as high proliferation index,was seen in recurrence group(DCIS and IC).FOXA1 expression is reduced across the groups though not statistically signifi cant.Conclusion:This is the first study to analyze novel transcription markers FOXA1 and GATA-3 in DCIS.Further work needs to be done on a larger cohort of DCIS cases with recurrence to better understand,which variables are best able to predict recurrence and guide therapy decision strategies.Maintenance of FOXA1 and GATA-3 expression in ER(-)DCIS may offer new promising targets for therapy in future.

Effect of Yiqi Tongyang Decoction(益气通阳方) on Blood T Cell Subsets in Patients with Chronic Immune Thrombocytopenia

Objective： To measure the proportions of blood T cel subsets, Th1, Th2, Th17, Th22, and Treg cel s, and other parameters in patients with chronic immune thrombocytopenia（CITP） before and after treatment with Yiqi Tongyang Decoction（益气通阳方, YTD） to explore T cel status of patients with CITP, and to define the mechanism of action of YTD. Methods： The changes in peripheral blood T lymphocyte subsets, and those of Th1, Th2, Th17, Th22, and Treg cel s in 30 patients with CITP（22 females and 8 males） were analyzed using multiparametric flow cytometry before and after treatment with YTD for 6 months, and 26 healthy volunteers（14 males and 12 females） acted as a control. T-box expressed in T-cel s（T-bet） and GATA binding protein 3（GATA-3） m RNA levels in patients and controls were analyzed using real-time reverse transcription-polymerase chain reaction. Results： The proportions of Th1, Th17, Th22, Th1/Th2, and Th17/Treg cells increased in the peripheral blood of patients with CITP compared to those in controls before YTD therapy（P〈0.05）. Th1 cel numbers and the Th1/Th2 ratio fel in the treated patients with CITP to approximate the values of the control group（P〉0.05）. Th17 cel numbers and the Th17/Treg ratio also decreased in the treatment group（P〈0.05）, but not to the levels of the controls. The number of Treg cel s in the peripheral blood of patients with CITP before treatment was lower than that in the control group（P〈0.05）, but increased after YTD treatment（P〈0.05）, but not to the level of controls. T-bet and GATA-3 m RNA levels in peripheral blood were initially higher in patients before treatment than controls（P〈0.05）, but decreased after YTD therapy（P〈0.05）. Conclusions： Imbalances in T lymphocyte levels, particularly those of Th1/Th2 and Th17/Treg cel s, play important roles in the pathogenesis of CITP. YTD efficiently regulated the dynamics of Th1/Th2 and Th17/Treg equilibria.