Aim To investigate the expression of GRKs in rats of synovial tissue with collagen-induced arthritis(CIA) and the effect of total glucosides of paeony.Methods SD rats were divided into six groups including normal,model,TGP(25,50,100 mg?kg-1)groups,GTW(40 mg?kg-1)group.Chicken type Ⅱ collagen was used to induce CIA in rats.The expression of GRKs was detected by Westernblot.Results The expression of GRK2,5,6 increased in model group than that in normal group.Compared with the model group,the expression of GRK2,5,6 decreased in TGP groups.Conclusion In rats of synovial tissue with CIA,the expression of GRKs was abnormal,and TGP could change the variation of GRKs which may be one of the mechanisms of TGP improvement CIA.

Aim To investigate the regulatory effects of Paeoniflorin on G protein-coupled signaling by synoviocytes of collagen-induced arthritis (CIA) rats. Methods CIA was induced by chicken type Ⅱcollagen. Pae (25, 50, 100 mg?kg-1) was administered to CIA rats from d 16 to d 23 after immunization. Arthritis was evaluated by hind paw swelling and arthritis index. Expression of inhibitory G protein (Gi) was detected by Western blotting technique. The level of cAMP in synoviocytes was measured by radioimmunoassay. Protein kinase A (PKA) activity was assessed by Kinase-Glo Luminescent Kinase Assay. Results There was remarkably secondary inflammation in CIA rats. The expression of Gi in synoviocytes increased. While cAMP level and PKA activity of synoviocytes decreased. The inflammatory responses of CIA rats were inhibited after administration of Pae. At the dose of 100 mg?kg-1, Pae reduced Gi expression. cAMP level and PKA activity were enhanced by Pae at the doses of 50 and 100 mg?kg-1 respectively. In ivtro, Pae (2.5, 12.5, 62.5 mg?L-1) reduced Gi expression, but enhanced the level of cAMP and PKA activity. Conclusion G protein-coupled signaling was associated with the pathogenesis of synovitis in CIA rats. Pae has anti-inflammatory effects on CIA rats by modulating G protein -coupled signaling.

Objective To use the nested PCR technology to diagnose and identify a case of imported Plasmodium ovale wallikeri subspecies .Methods Blood sample was collected from 1 case of initially diagnosed imported tertian malaria and performed the ex‐amination of microscopy ,rapid diagnostic tests (RDTs) and nested‐PCR .Moreover the sequencing was conduced .Results RDTs showed the negative result ;the ring form and trophozoite of Plasmodium could be observed in the blood smear by microscopy ;the Plasmodium ovale wallikeri subspecies specific primer rOVA 1v/rOVA2v was adopted for conducting nested PCR ,the specicific am‐plification band appeared at 760 bp ,after sequencing and Blast aligning ,its coincidence with the partial sequence of Plasmodium ovale wallikeri subspecies in the Genbank database was 99% .Conclusion This patient is the first case of Plasmodium ovale wallik‐eri subspecies infection in Hubei province by nested PCR and sequencing analysis .

Objective:To establish the methods of isolated culture and functional identification of mice bone marrow derived tolerogenic dendritic cells (CD11b+F4/80 +TDCs) in vitro.Methods: Mice bone marrow cells were isolated and cultured to obtain iDCs with the simulation of mouse rmGM-CSF and rmIL-4.CD11b+F4/80 +TDCs were purified by fluorescence-activated cell sorting on day 6.The morphological changes of TDCs were observed with the inverted microscope dynamically .The expression of CD11b+F4/80 +TDCs were analyzed by the flow cytometry .Tolerogenic function of CD11b+F4/80 +TDCs was evaluated by the expression of MHCⅡ, CD83, IDO, TLR-2, IL-10 and TGF-β1.The expression of MHCⅡ was analyzed by the flow cytometry , and the expression of CD83, IDO and TLR-2 were analyzed by immune-histochemistry.The levels of IL-10 and TGF-β1 in the supernatant of CD11b+F4/80 +TDC were analyzed by ELISA .Meanwhile mature DCs ( mDCs) induced by LPS were used as control .Results:The fresh isolated bone marrow cells look like round and small under microscope .After two days of culture , cells became big and formed into clusters . Five or six days later, cells clusters increased, and the morphology of cells became irregular .At the same time, more dendrite ap-peared on the surface of cells .The percentage of CD11b+F4/80 +TDCs induced by rmGM-CSF and rmIL-4 was about 23%, and the purity of the purified BM CD11b+F4/80 +iDC was about 99%.Compared with mDCs, CD11b+F4/80 +TDCs expressed low levels of MHCⅡand CD83 and high levels of IDO, TLR-2, IL-10 and TGF-β1.Conclusion:CD11b+F4/80 +TDCs derived from mouse bone marrow could be induced successfully by rmGM-CSF and rmIL-4 in vitro.CD11b+F4/80 +TDCs showed tolerogenic function by the expressions of IL-10, TGF-β1, IDO and TLR-2.

Objective To understand the malaria epidemic situation in Hubei Province from 2010 to 2014,so as to provide the evidence for formulating the effective malaria elimination strategies and measures in this province. Methods The data from the Disease Reporting Information System of Chinese Center for Disease Control and Prevention were collected and analyzed with the descriptive epidemiological method for the epidemiological characteristics of malaria in Hubei Province from 2010 to 2014. Results A total of 997 malaria cases were reported in Hubei Province from 2010 to 2014,there were 618 cases of vivax malaria,352 cases of falciparum malaria,18 cases of Plasmodium ovale infection,and 9 cases of Plasmodium malariae infec?tion. Among all the reported cases,479 were local cases and 518 were imported cases. No local malaria cases were reported from Hubei Province since 2013. The overall imported malaria cases showed a gradual increasing trend from 2010 to 2014,the pro?portion of falciparum malaria increased quite significantly from 2010 to 2014. The malaria cases were mainly distributed in Xiangyang,Wuhan,Xiaogan,Yichang,Jingmen and Suizhou cities,reaching 81.85%of the cases of the whole province. There were 810 male cases and 187 female cases,with a sex ratio of 4.33∶1. The local malaria cases were mainly aged from 40 to 69 years,accounted for 78.29%of the total local cases,and 88.22%(457/518)of the whole local cases were concentrated in 20-49 age groups. The local cases were mainly farmers(67.01%). Among the imported malaria cases,the occupation distribution concentrated mainly on the worker,migrant worker,and farmer(63.90%). Conclusions The local malaria epidemic situation has been effectively controlled in Hubei Province,which reflects the initiative achievements of malaria elimination. However, there are still many imported malaria cases from abroad. Therefore,the imported malaria from abroad still remains the key of ma?laria control in Hubei Province.

Objective To evaluate the effectiveness of the application of Wondfo Rapid Diagnostic Kit (RDTs) in the diagnosis of imported malaria cases in the Malaria Reference Laboratory of Hubei Provence. Methods The complete blood samples of malaria cases and negative card deletion cases reported in Hubei Province from January 2015 to June 2018 were collected and retrospectively analyzed. The results of the provincial malaria reference laboratory were used as the standard, and were compared with those results detected by RDTs, microscopic examination and nested PCR. The differences were statistically analyzed. Results A total of 440 complete samples were collected by the Malaria Reference Laboratory of Hubei Provence, of which 418 samples were confirmed as positive, and 22 samples were confirmed as negative. In terms of the identification ability of P. falciparum, RDTs performed the best, with a coincidence rate of 100.00%, and the coincidence rates nested PCR and microscopic examination were 97.49% and 91.40%, respectively. In terms of the identification specificity for another 3 species of Plasmodium (P. vivax, P. ovarian and P. vivax), nested PCR was the best, the microscopy method was the second best, and RDTs was the lowest. Based on the comprehensive analysis of 12 individual indicators, RDTs had the highest score (32), while the microscopic examination and nested PCR scored 24 and 19, respectively. Conclusion RDTs had certain advantages in the detection of malaria, but they had a low identification specificity for different species. Thus, they can be used as auxiliary tools for microscopic examination and widely used in surveillance work after malaria elimination in Hubei Province.

Objective To understand the number and prevalence of chronic filariasis patients in Hubei Province, and to provide a basis for effective care for patients. Methods The chronic filariasis patients registered in Hubei Province were interviewed to investigate the general condition of patients, the results of previous microfilaria examinations, and the current clinical symptoms, signs and onset of illness. Results A total of 487 chronic filariasis patients were investigated. The youngest was 32 years old, the oldest was 97 years old, and the average age was 76 years old. The 80-89 years old group had the largest number, accounting for 44.97%. The male accounted for 46.61%, and the female accounted for 54.39%. The occupation was dominated by farmers, accounting for 89.94%. Patients with lymphangitis accounted for 72.28%, with dermatitis accounted for 89.12%, with chyluria accounted for 11.29%, and with hydrocele accounted for 6.16%. Among the 352 patients with lymphangitis/lymphadenitis, the site of the attack was mainly at groin and lower limbs, accounting for 96.02%. Among the 433 patients with lymphedema/elephantiasis, the disease staging of the left and right legs was mainly concentrated in stage I-III, and there was no stage VII patient. Among the 55 patients with chyluria, the dysuria accounted for 30.91%. Of the 30 patients with hydrocele, 17 had tenderness, and 8 had positive light transmission tests. Conclusions Chronic filariasis patients in Hubei Province gradually decreased with the natural attrition. More care should be given by CDC at all levels and primary medical units to patients with chronic filariasis to alleviate their symptoms and improve their quality of life. Since the filariasis has not been eliminated globally, it is still necessary to strengthen the surveillance of migrant workers who go abroad to filariasis-endemic areas.