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.

Objective To investigate the protective effect of breviscapine on ischemia-reperfusion renal injury, which provides scientific theoretical basis for clinical prevention and treatment of renal ischemia-reperfusion injury. Method 36 SPF male healthy SD rats were randomly divided into 3 groups, 12 rats in each group. Group A was ischemia-reperfusion group, group B was erigeron breviscapus injection preconditioning group, and group C was sham operation group. Rats in group A and C were injected with normal saline, while rats in group B were given 12 ml/kg erigeron breviscapus injection by intraperitoneal injection., After 14 days, renal function, renal antioxidant indexes, renal cell apoptosis indexes and expression of Bcl-2, Bax in renal tissue of three groups were compared. Results The indexes of renal function showed that blood urea nitrogen (BUN) and Serum creatinine (SCr) in group B were significantly lower than group A (P<0.05), but significantly higher than group C (P<0.05). Renal antioxidant indexes showed that superoxide dismutase (SOD) and malondialdehyde (MDA) in group B were significantly higher than group A (P<0.05), but significantly lower than group C(P<0.05). The index of renal cell apoptosis showed that the apoptosis index AI in group B was significantly lower than group A (P<0.01), but significantly higher than group C (P<0.01). The results of immunohistochemical staining showed that the expression of Bcl-2 in group B were significantly higher than that of group A and group C (P<0.05), and the expression of Bax in group B was significantly lower than that in group A (P<0.05) while significantly higher than group C (P<0.01). The ratio of Bcl-2/Bax in group B was significantly higher than group A and group C (P<0.05). Conclusion Erigeron Breviscapus Injection may have a protective role on renal ischemia reperfusion injury though antioxidant and anti-apoptosis.

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 analyze the malaria control measures and epidemic trend in Hubei Province from 1974 to 2015, so as to provide the evidence for malaria elimination path analysis. Methods The malaria control data in Hubei Province from 1974 to 2015 were collected and analyzed retrospectively by descriptive epidemiological methods. Results The epidemic pro?cess of malaria in Hubei Province was divided into four stages. From 1974 to 1979,it was high prevalence state of malaria,and the average annual incidence was 174.47/10 000. From 1980 to 1999,the main control strategies were to control the infection source and mosquitoes,and the average annual incidence was 17.30/10 000,significantly downward. From 2000 to 2009, through the surveillance of infection sources and controlling malaria outbreaks and strengthening the floating population manage?ment,the average annual incidence was 0.42/10 000. After 2010,followed by the elimination phase of malaria,the incidence continued to decline. In 2013,there was no local infection for the first time. The difference of average annual incidence among above?mentioned stages was statistically significant(χ2=1 254.36,P<0.05). Conclusions The malaria epidemic process in Hubei Provincial experienced the high epidemic stage,sharply drop stage,low incidence phase and the elimination phase. How?ever,the natural factors affecting malaria still exist. Therefore,strengthening the control of imported malaria and surveillance should be the main task in the process of eliminating malaria in the future.

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 study the effect of combined acupuncture and rehabilitation training on early-stage Barthel index of ischemic stroke cases.Method:Fifty ischemic stroke cases were randomized into a treatment group of 24 cases and control group of 26 cases.In addition to routine internal treatment methods,rehabilitation training was applied to cases in the control group,and combined acupuncture and rehabilitation training were applied to cases in the treatment group.Warming and reinforcing manipulations were applied to Jianyu (LI 15),Quchi (LI 11),Hegu (LI 4),Yanglingquan (GB 34),Yinlingquan (SP 9),Zusanli (ST 36),and Sanyinjiao (SP 6),20 min of needle retaining for each point in one treatment.Then the scores of clinical neurological impairment and Barthel index of stroke cases were evaluated before the treatment and 3 weeks after respectively.Results:The significant progress and progress cases in the treatment group were 14 and 12 respectively,whereas that in the control group were 6 and 11 respectively,coupled with 7 cases of no progress.The therapeutic effect in the treatment is substantially better than the control group (P＜0.05).There was no significant difference (P＞0.05) in neurological impairment and scores of Barthel index of the two groups before the treatment,whereas significant difference (P＜0.05) after the treatment.The scores of neurological impairment in the treatment group were significantly reduced after the treatment (P＜0.05),and the scores of Barthel index were significantly increased (P＜0.05).Conclusion:Combined acupuncture and rehabilitation training can produce significantly better effect on early neurological impairment and Barthel index of stroke cases than rehabilitation training alone.

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 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.

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.