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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.
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Sepsis has the characteristics of high morbidity and high mortality, which is the main cause of death in critically ill patients. Peripheral 5-hydroxytryptamine (5-HT) is mainly synthesized and secreted by enterochromaffin cells and plays a role in the development and progression of inflammation. This article focuses on 5-HT, 5-HT transporter (SERT) and selective serotonin reuptake inhibitors (SSRIs) in immune response, inflammatory factor release, oxidative stress, intestinal bacterial translocation and microcirculation of sepsis. The role of the review is to find new ideas for the clinical treatment of sepsis.
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Objective To evaluate the effect of a protein kinase CK2 inhibitor on intracellular levels of reactive oxygen species and DNA double?stand break in human non?small cell lung cancer H460 cells. Methods H460 cells were exposed to 0, 12?5, 25.0, and 50.0μmol/L quinalizarin, a specific inhibitor of protein kinase CK2, for 24 hours. The changes in protein and mRNA levels of CK2 subunits were measured. Flow cytometry was used to measure changes in the intracellular levels of reactive oxygen species in H460 cells after 4 or 24 hours of quinalizarin treatment. Immunofluorescence assays were performed to determine the effect of the CK2 inhibitor onγ?H2 AX expression and the average fluorescent number ofγ?H2 AX foci in H460 cells. Comparison was made by analysis of variance and t test. Results There were no significant differences in protein or mRNA levels of CK2 subunits in H460 cells after quinalizarin treatment ( CK2α,0μmol vs. 12?5 μmol/L, P=0?966;0 μmol/L vs. 25 μmol/L, P=0?355;0 μmol/L vs. 50 μmol/L, P=0?864, CK2α’ , 0 μmol/L vs. 12?5μmol/L,P=0?409;0μmol/L vs. 25μmol/L,P=0?833;0μmol/L vs. 50 μmol/L, P=0?0. 746, CK2β, 0 μmol/L vs. 12?5 μmol/L, P=0?532;0 μmol/L vs. 25 μmol/L, P=0?830;0 μmol/L vs. 50 μmol/L, P= 0?061 ) . The intracellular levels of reactive oxygen species were substantially elevated in H460 cells with the increase in quinalizarin concentration and treatment time. Different concentrations of quinalizarin resulted in dose?and time?dependent increases in the numbers of γ?H2 AX foci after 4 and 24 hours of treatment ( treated by Quianlizarin for 4 or 24 h, 0 μmol/L vs. 12?5μmol/L,12?5 μmol/L vs. 25 μmol/L, 25 μmol/L vs. 50 μmol/L, all P=0?000, concentration is 12?5μmol/L,25 μmol/L or 50 μmol/L, 4 h vs. 24 h, all all P=0?000 ) . Conclusions Quinalizarin can increase the intracellular levels of reactive oxygen species and DNA double?stand break in H460 cells by inhibition of protein kinase CK2 activity. This study provides a theoretical basis for using quinalizarin as a potential radiosensitizer for lung cancer.
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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.
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OBJECTIVE:To establish a method for determining the content of paeoniflorin in Xiaohe syrup. METHODS:HPLC method was adopted. The Waters SunFire C18 column was used with the mobile phase of acetonitrile-0.1% phosphoric acid(12∶88, V/V)at the flow rate of 1.2 ml/min;the detection wavelength was 230 nm with column temperature at 30℃. Sample size was 10μl . R E-SULTS:There was a good linear relationship between the amount of paeoniflorin and peak area in the range of 0.061 0-1 . 2 2 0 8 μg (r=0.999 9). The RSDs of precision,stability and reproducibility tests were all less than 2% and the average recovery was 98.77%(RSD=1.09%,n=6). CONCLUSIONS:The method is simple,rapid and accurate and can be used for the content deter-mination of paeoniflorin in Xiaohe Syrup.
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Objective To explore the effect of SFI in radiation-induced mice brain injury after 20 Gy cranial radiation.Methods The mice were divided into three groups:(1) control group,(2) RT-only group:the whole brain was irradiated with a dose of 20 Gy,(3) RT and SFI group:SFI at 20 ml/kg/d from 4 weeks after 20 Gy cranial radiation theraty(CRT).Results Morris water maze test showed that the latency of the irradiated group was longer than control group and SFI improved the cognitive function of mice (t =6.34,6.70,P ＜0.05).The expression of TNF-α reached to the highest level at 3 h after irradiation,and then it decreased but got the second higher level again at 4 weeks after irradiation.The expression of IL-1 β reached to the highest level at 72 h after irradiation and decreased until 4 weeks after irradiation.SFI decreased both expressions of TNF-α (t =11.34,9.70,6.07,P ＜ 0.05) and IL-1 β (t =12.27,5.70,7.52,P ＜ 0.05).Immune florescence staining showed that SFI reduced the number of activated microglia (t =12.35,8.64,7.82,P ＜ 0.05)and inhibited the translocation of p65 of microglia after irradiation.Conclusions Findings suggest that SFI may decrease microglial activation and suppress the expression of TNF-α and IL-1β by inhibiting the translocation of NF-κB p65 and then attenuate irradiation-induced brain injury.
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Objective To investigate the changes of hippocampal neurogenesis and cognitive dysfunction induced by cranial radiation therapy.Methods C57BL/6J mice aged 10 d were subjected to 10 Gy whole brain irradiation with 6 MV X-rays to develop irradiation-induced brain injury model.Morris water maze was designed to estimate spatial learning and memory.At different time post irradiation,brain tissue was removed to stain with hematoxylin-eosin for the pathological results.DCX and PCNA immunohistochemical staining was used to mark the level of neurogenesis in the hippocampus,and ED1immunohistochemical staining to mark the activation of microglia.The TUNEL assay was used to assess the apoptotic neuron death in situ in the hippocampus.Real-time PCR was supplied to inspect the expression of TNF-α and IL-1 β mRNA.Enzyme Linked Immunosorbent Assay (ELISA) was tested for the concentration of TNF-αt in the plasma.Results Pathological studies demonstrated that radiation could induce interstitial edema,inflammatory cell infiltration,cell degeneration,necrosis,apoptosis in the acute phase,edema subsiding,reduction of inflammatory cells,and cytothesis in the dentate gyrus of the hippocampus.IHC studies revealed that,at different time post irradiation,the number of DCX-positive cells and PCNA-positive cells decreased (F =4.9-12.5,5.2-15.7,P ＜ 0.05) but ED1-positive cells increased significantly (F =20.8,P ＜ 0.05).TUNEL-positive cells began to appear in the dentate gyrus of hippocampus 6 h post-irradiation,and its number reached to the highest level at 48 h post-irradiation (F =15.1,P ＜ 0.05).The formation of γ-H2AX foci got at the top 0.5 h post-irradiation (F =18.4,P ＜0.05) and then decreased.After irradiation,the expressions of TNF-α and IL-1β mRNA in the the irradiated group was higher than those of the control group (t =16.3,12.7,P ＜ 0.05).The concentration of TNF-α in the plasma of the irradiated group was higer than that in the control group 3 h post-irradiation,and maximized at 1 week post-irradiation (F =10.5,P ＜ 0.05).Morris water maze tests showed that the latency had no significant differences between the irradiated group and the control group at 1,2,3 d postirradiation,but the latency in the irradiated group was longer than that in the control group with a significant differences at 4,5,6 d post-irradiation (F =7.01,8.17,4.22,P ＜ 0.05).Conclusions Irradiation-induced cognitive dysfunction may be caused by microglial activation and suppression in hippocampal neurogenesis following cranial radiation therapy.
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The present study investigated the enhanced radiosensitivity of U-251 cells induced by sodium butyrate (NaB) and its possible mechanisms. Increased radiosensitivity of U251 cells was examined by clonogenic cell survival assays. The expression of Ku70 mRNA and protein was detected by using RT-PCR and Western blotting respectively. γ-H2AX foci were measured at different time points after ionizing irradiation alone or combined with NaB treatment. The results showed that cell survival rate was significantly reduced, both D0 and Dq values were decreased (D0: 1.43 Gy vs. 1.76 Gy; Dq: 1.22 Gy vs. 2.05 Gy) after the combined treatment as compared with irradiation alone, and sensitivity enhancing ratio (SER) reached 1.23. The average number of γ-H2AX foci per cell receiving the combined treatment was significantly increased at different time points, and the expression levels of Ku70 mRNA and protein were suppressed by NaB in a dose-dependent manner. It was concluded that enhanced radiosensitivity induced by NaB involves an inhibited expression of Ku70 and an increase in γ-H2AX foci, which suggests decreased ability in DSB repair.
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Objective To explore the inhibitory effects of Tanshinone Ⅱ A on the radiationinduced microglia activation and the possible mechanism.Methods Microglia cells BV-2 were irradiated with 2,4,8,16,and 32 Gy doses or sham-irradiated in presence or absence of 1.0 μg/ml Tanshinone Ⅱ A for 12 h,respectively.The effects of Tanshinone Ⅱ A on radiation-induced pro-inflammatory cytokines were evaluated using real-time PCR.The expression level of NF-κB p65 in cytoplasm and nucleus was measured by using Western blot.Immunofluorescence staining and confocal microscopy analysis were applied to detect the expression of γ-H2AX and p65 post-irradiation.Results The microglia cells were activated at 16,32 Gy post-irradiation.Radiation-induced release of the pro-inflammatory cytokines in BV-2 cells was detectable after irradiation.Tanshinone Ⅱ A decreased radiation-induced release of proinflammatory cytokines(t=5.56,P ＜ 0.05).Furthermore,western blotting showed that Tanshinone Ⅱ A could attenuate the nuclear translocation of NF-κB p65 submit post-irradiation.Immunofluorescence staining showed that γ-H2AX foci formation while p65 translocation into nucleus post-irradiation.Conclusions Tanshinone Ⅱ A exerts anti-inflammatory properties by suppressing the transcription of proinflammatory cytokine genes that might be associated with NF-κB signaling pathway.It is postulated that irradiation causes immediate cellular reaction and DSB triggers the molecular response which leads to NFκB pathway activation.
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Objective To explore the inhibitory effects of Corilagin on the production of proinflammatory cytokines in microglia induced by radiation. Methods The cytotoxicity of Corilagin was measured by MTT assay. Microglia BV-2 cells were irradiated 0 or 32 Gy after pretreated with Corilagin for 12 hours. Realtime-PCR was used to detect the mRNA levels of inflammatory cytokines, such as IL-1β,TNF-α on several time-points. The content of nitric oxide (NO) was determined with nitrate reductase method. The translocation of NF-κB was measured by Western blot and immunocytochemical stain.Confocal microscopy was used to observe the expression of Iba-1 and Nemo. Results No cytotoxicity was detected on BV-2 cells with 1-10 μg/ml Corilagin. Iba-1 expression in microglia cells was activated by irradiation, the expression levels of inflammatory cytokines, such as IL-1β, TNF-α and NO were also elevated. Whereas, the production of IL-1 β, TNF-α in activated microglia cells was significantly inhibited with 5 μg/mL corilagin ( tIL-1β = 6. 341, tTNF-α = 3.41 1, tNO = 3. 134, P ＜ 0. 05 ). Corilagin significantly inhibited the expression of Nemo and the translocation of NF-κB p65. Conclusion Corilagin could inhibit the activation of irradiated microglia cells and down-regulate the expression of inflammatory cytokines, via inhibition of the NF-κB signaling pathway.
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Objective To investigate ff the γ-H12AX foci is a precise index for the DSB formation and repair in mature neurons of brain in vivo after clinically relevant doses irradiation. Methods For the DSB formation experiment, the mature neurons in the neocortex of brain tissue of C57BL/6 mice were analyzed at 10 rain after whole-body irradiation with 0.1, 0.5 and 1.0 Gy. For the DSB repair kinetics experiment, the mature neurons in the neocortex of brain tissue of repair-proficient (C57BL/6 mice) and repair-deficient mouse strains (BALB/c, A-T and SCID mice) were analyzed at 0.5, 2.5, 5, 24 and 48 h after whole-body irradiation with 2 Gy. The mature neurons in the neocortex of brain tissue of sham-irradiated mice of each strain served as controls. γ-H2AX immunohistochemistry and γ-H2AX and NeuN double immunofluorescence analysis was used to measure DSBs formation and repair in the mature neurons in the neocortex of brain tissue of the different mouse strains. Results For the DSB formation experiment, γ-H2AX foci levels with a clear linear dose correlation and very low backgrounds in the nuclei in the neocortex of brain tissue were observed. Scoring the loss of γ-H12AX foci allowed us to verify the different, genetically determined DSB repair deficiencies, including the minor impairment of BALB/c mice. Repair-proficient C57BL/6 mice exhibited the fastest decrease in foei number with time, and displayed low levels of residual damage at 24 h and 48 h post-irradiation. In contrast, SCID mice showed highly increased γ-H2AX foci levels at all repair times (0.5 h to 48 h) while A-T mice exhibited a lesser defect which was most significant at later repair times (≥ 5 h). Radiosensitive BALB/c mice exhibited slighdy elevated foei numbers compared with C57BI./6 mice at 5 h and 24 h but not at 48 h post-irradiation. Conclusion Quantifying the γ-H2AX loci in normal tissue represents a sensitive tool for the detection of induction and repair of radiation-induced DSBs at clinically relevant doses in vivo.
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Objective To evaluate the current sanitary situation of the rural household latrine and the effects of decontamination of the feces in some epidemic areas of schistosome in Hubei province.Methods Six villages(4 villages with the sanitary latrines,2 villages with the non-sanitary latrines) were chosen for the investigation and 30 latrines were selected randomly in each village in September of 2007.The structure of the latrine and the situation of the faecal treatment were investigated by questionnaires.At the same time,the feces samples were collected and detected according to the related sanitary standards.Results The rate of the sanitary latrines without maggots,pupas and flies was 95%,while the rate of the non-sanitary latrines with the same situation was 33.33%.After non-hazardous treatment in the three-case-cesspool and the firedamp latrines,the qualified rates of the roundworm eggs were 95.6% and 98.6% respectively;the qualified rates of fecal coliform were both 100%;no schistosome eggs,the alive schistosome eggs and the alive roundworm eggs were detected.The morbidity rate of schistosomiasis of the villages with the sanitary latrines was 43.7%,lower than those with the non-sanitary latrines.Conclusion The faecal treatment of the rural household latrine in epidemic areas of schistosome can improve the sanitation and kill the pathogens in faeces effectively,and also can decrease the morbidity rate of schistosomiasis obviously.
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To explore the effects of Taushinone ⅡA on the proliferation, apoptosis and gene ex- pression of p53 and bcl-2 in human gastric carcinoma MKN-45 cells. Cell count and MTT assay were used to study the proliferation-inhibiting effect of Tanshinone ⅡA on MKN-45 cells. The effect of Tanshinone ⅡA on the cell cycle and apoptosis of MKN-45 cells were examined by propidium io- dide (PI) staining and flow cytometry. Semi-quantitative RT-PCR was used to further verify the ex- pression of p53 and bcl-2 gene after exposure to Tanshinone ⅡA in MKN-45 cells. The results showed that Tanshinone ⅡA significantly inhibited the growth and proliferation of MKN-45 cells in a dose- and time-dependent manner (P<0.05). Tanshinone ⅡA arrested MKN-45 cells in G2/M phase which led to an obvious accumulation of G2/M phase cells while decreased number of G0/G1 phase cells. This resulted in apoptosis of MKN-45 cells and the apoptosis rate was as high as 43.91% after treatment with 2.0 μg/mL Tanshinone ⅡA for 96 h. It was also found that Tanshinone ⅡA up-regulated expression of p53 gene and down-regulated expression of bcl-2 gene. The cytostatic and antiproliferative effect of Tanshinone ⅡA makes it a promising anticancer agent for the treatment of gastric carcinoma.