Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
Ammonium Sulfate ; chemistry ; Glycerolphosphate Dehydrogenase ; chemistry ; isolation & purification ; Molecular Weight ; Polyethylene Glycols ; chemistry ; Water

Objective To analyze the epidemiological characteristics of measles in Jinshan District of Shanghai from 2004 to 2015 in order to provide guidelines for measles control. Methods A descriptive epidemiological analysis was conducted on measles cases reported in Jinshan District during 2004 to 2015. Epidemiological data of those cases were collected from surveillance system of infectious disease, Measles Surveillance System ( MSS) and case study records. Results A total of 225 confirmed measles cases were analyzed in this study with an annual incidence of 2. 66/100 000. The highest incidence was 11. 27/100 000 in 2005 and the lowest incidence was 0. 14/100 000 in 2011, taking on a descending trend (Z=-6. 689, P<0. 000 1). The number of cases from March to May accounted for 65. 33%. The incidence of measles in floating population (4. 00/100 000) was higher than that in resident population (2. 18/100 000) (χ2=20. 467, P<0. 000 1). The measles cases mainly concentrated in age groups of ≥20 years and <5 years. Workers, scattered children, domestic workers, farmers and fishermen had high incidences of measles. Mea-sles vaccine immunization history was clear in nine cases. Conclusion The incidence of measles in Jinshan District shows a general trend of decline in this decade with time and population clusters in occurrence, sug-gesting that the occurrence of measles in Jinshan is under control. It is necessary to strengthen measles sur-veillance, regular immunization among children and supplementary immunization among adults, especially in floating population, in order to maintain high levels of immune protection.

Objective: To investigate the effect of a comprehensive exercise intervention program combined with diet control on ANGPTL2 and vascular endothelial function in obese male adolescents, and to provide theoretual basis for exercise to reduce the risk of cardiovascular disease in obese male adolescents. Methods: Forty two obese male adolescents and 10 healthy male adolescents were selected, a comprehensive intervention of 6 weeks of exercise combined with diet control was carried out on obese male adolescents, and changes in morphological function, glucose and lipid metabolism, inflammation level and vascular endothelial function were measured before and after the intervention. Results: After 6 weeks of comprehensive intervention, the morphological and functional indicators of obese adolescents were significantly reduced:weight, BMI, waist circumference, waist to hip ratio, systolic blood pressure, diastolic blood pressure ( P <0.05); blood lipid levels significantly decreased, including TC, TG and LDL-C ( P <0.05); inflammation levels significantly decreased, including TNF α and ANGPTL2 ( P <0.05); Vascular endothelial function significantly improved:RHI, NO/ET-1, NO, AI( P <0.05). Before and after 6 weeks of comprehensive intervention, the ANGPTL2 of obese adolescents was significantly correlated with weight ( r =0.37), BMI ( r =0.45), RHI ( r =-0.46), NO/ET-1 ( r =-0.41), NO( r =-0.45), and AI ( r =0.33)( P <0.05). Conclusion The comprehensive intervention can effectively improve the morphological function, blood lipid level and vascular endothelial function of obese male adolescents, and reduce the circulating level of ANGPTL2. ANGPTL2 may be involved in the process of comprehensive intervention to improve the vascular endothelial function of obese adolescents.

ObjectiveTo investigate the application of intraoperative Computed Tomograph (CT) using in surgery for complex acetabular fractures.MethodsFrom June 2008 to December 2010,14 patients (9 males,5 females; with the mean age of 45.1 years; range,28-62 years) with complex acetabular fractures were operated using intraoperative CT.Preoperative radiotherapy and CT scan were adopted to evaluate the fractures.Three dimensional reconstruction based on CT scan was used to mimic surgery.The surgery approach and the type of internal fixators were noted.Intraoperative C-arm and CT scan were used to evaluate the fractures reduction respectively.Decision of additional reduction was made by surgeons according to above mentioned methods respectively and the results were noted.Comparing to preoperative design,the change of surgery plan were noted.Overall time,frequency and radiation dose of intraoperative CT scan were also noted.ResultsAll patients in this study received average 2.7 times of intraoperative CT scan.Mean time of CT scan was 40.4 min and the overall dose of radiation was 47.2 mGy.Decision of additional reduction was made in 3 cases according to C-arm radiography and 4 cases according to CT scan (above mentioned 3 cases were included).The change of surgery plan was made in one case.In postoperative radiography evaluation according to Matta's score system,anatomical reduction were achieved in 8 cases,imperfect reduction in 3 cases and poor reduction in 3 cases.ConclusionIntraoperative CT scan increases the radiation time and dose of patients dramatically.When used to evaluate fracture reduction intraoperatively,it can't take the advantage of traditional C-arm radiography.When delicate preoperative plan is made with radiography and three dimensional reconstruction based on CT data,the efficiency of intraoperative CT scan for complex acetabular fractures are to be discussed.

Objective To analyze the imaging characteristics of lung cancer detected by LDCT ( low-dose CT )lung cancer screening. Methods Between July 1st, 2007 and June 30th, 2013, 7 141 asymptomatic enrolled participants aged 40-88 years old (male 4 710, female 2 431, median age 47), and 1 071 volunteer participants aged ≤39 underwent chest LDCT. The imaging characteristics were analyzed retrospectively in lung cancer pathologically proved. Three types were classified according to the imaging findings: solid lesion, part-solid lesion and non-solid lesion. Results A total of 31 participants (32 lesions) were diagnosed as lung cancer, including 30 adenocarcinomas, 1 carcinoid and 1 small cell lung cancer. The detecting rate of the lung cancer was 0.4%(31/8 212). The solid lesion was further classified as classical solid nodule, irregular solid lesion and atypical solid nodule, and the part-solid lesion was further classified as part-solid nodule, irregular part-solid lesion and cystic part-solid nodule. Lung cancer or probably lung cancer was diagnosed in 24 cases (77.4%), and uncertainty diagnosis was made in 3 cases (9.7%). Benign or probably benign was diagnosed in 3 cases, and another 1 cases were missed at baseline screening. The false positive rate and the false negative rate was 9.7%and 3.1%, respectively. Conclusion The imaging characteristics of lung cancer detected by LDCT are varied, which provide preliminary experience in lung cancer screening.

Objective: To investigate the effects of exercise combined with dietary intervention on glucose and lipid metabolism, liver and kidney function and cardiovascular function in obese adolescents. Methods: Seventy-one obese adolescents were enrolled in this study. Six weeks of exercise combined with dietary intervention were performed to determine anthropometry, glucose and lipid metabolism, liver and kidney function and cardiovascular function before and after intervention. Results: After 6 weeks of comprehensive intervention, in addition to significant improvement in anthropometry indicators, fasting insulin [(18.76±11.46 vs 11.32±6.54)uU/L], LDL-C [(2.96±0.69 vs 2.22±0.62)mmol/L], TG [(1.57±0.82 vs 0.89±0.37)mmol/L] and TC [(4.52±0.76 vs 3.53±0.62)mmol/L] decreased significantly and insulin resistance improved significantly. AST [(36.1±32.28 vs 22.89±7.27)U/L], ALT [(57.42±61.25 vs 27.86±22.12)U/L], blood urea nitrogen [(4.78±0.89 vs 3.44±0.79)mmol/L] and uric acid [(498.83±120.6 vs 471.07±120.96)mmol/L] were significantly decreased and the detection rates of fatty liver and kidney were significantly decreased(P<0.05), RHI(1.34±0.28 vs 1.69±0.45) and nitric oxide/ endothelin-1 (ET-1) (1.69±0.41 vs 2.67±0.86) significantly increased and atherogenic index (3.06±0.96 vs 2.58±0.80) significantly decreased(P<0.01). Conclusion The comprehensive intervention of exercise combined with diet can effectively improve anthropometry indicators, glycolipid metabolism, liver and kidney function as well as cardiovascular function of obese adolescents.

The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.

OBJECTIVE Despite the status of cisplatin (DDP) as a classical chemotherapeutic agent in the treatment of cancer, the development of multidrug resistance often leads to a failure of DDP therapy.Traditional Chinese medicine(TCM)as adjuvant chemotherapy of cancer drugs in China has been widely used in cancer treatment.ZuoJin WAN (ZJW),a TCM formula,was proved reversing drug resistance in gastric cancer,but its exact mechanism was still unclear. METHODS CCK-8 assay was used to detect the cell viability. The levels of proteins and mRNA were evaluated using Western blot and q-PCR. Mitochondrial membrane potential was measured by fl ow cytometry. Depolymerisa-tion of F-actin and translocation of G-actin(gamma-actin)from the cytoplasm to the mitochondria was detected using an immuno fl uorescence assay. RESULTS phosphorylated coflin-1 (p-coflin-1) was overexpressed in the DDP-resistant human gastric cancer cell lines SGC7901/DDP and BGC823/DDP, relative to the respective parent cell lines(SGC7901 and BGC823),and DDP induced the dephosphory-lation of p-coflin-1 in both parent lines but not in the DDP-resistant lines. However, ZJW could induce the dephosphorylation of pcoflin-1 and promote coflin-1 translocation from the cytoplasm into the mito-chondria in both SGC7901/DDP and BGC823/DDP cells. This mitochondrial translocation of coflin-1 was found to induce the conversion of flamentous actin to globular-actin, activate mitochondrial dam-age and calcium overloading, and induce the mitochondrial apoptosis pathway. These effects of ZJW on DDP-resistant human gastric cancer cell lines could be reversed via transfection with coflin-1-specifc siRNA,or treatment with a PP1 and PP2A inhibitor.CONCLUSION ZJW can be used as an inhibitor of chemoresistance in gastric cancer, which may partly be due to dephosphorylation of p-coflin-1 via the activation of PP1 and PP2A.

Objective To analyze the spatial point pattern distribution characteristics of hemorrhagic fever with renal syndrome (HFRS) in Jingzhou city, Hubei province during the two seasons spring- summer and autumn-winter of 2017, to discuss its high incidence area and reason, and to provide basis for the resource allocation of public health. Methods The analytical data was collected from Infectious Disease Reporting Information System in China, and the spring-summer season was from March to August of 2017, while the autumn-winter was from the September of 2017 to the February of 2018. The Ripley's K-function and kernel density estimation were applied to analyze the spatial point pattern distribution and compare the distribution characteristics of spatial point pattern between the two seasons. Results In 2017, 133 cases of HFRS were reported in Jingzhou city, including the spring- summer and autumn-winter two pick incidences. The strongest aggregation distance was 17.77km in spring-summer season, and 14.40 km in autumn-winter season. The spatial gathering center was located in the north of Jianli County in spring-summer, and it moved to the south of Jiangling County and Shashi District in autumn-winter. Conclusions The key areas for the prevention and control of HFRS in Jingzhou City are Jiangling County, the southern part of Shashi District and the northern part of jianli county. The key groups are the residents of the urban-rural junction in the southern part of Shashi City, residents along the route of large-scale projects, and farmers engaged in agricultural planting or crayfish breeding in the gathering areas.

Objective:To evaluate effect of diammonium glycyrrhizinate(DG)on lipopolysaccharide(LPS)-induced TLR4/NF-κB inflammatory signaling pathway in BV2 microglial cells and to explore its potential regulatory mechanisms on ameliorating neu-roinflammation.Methods:BV2 microglia injury model was induced by LPS and treated with 20 and 60 μmol/L DG.MTS was used to determine vitality of cells.NO level secreted by cells was detected by Griess.Inflammatory factors TNF-α,IL-6,and IL-1β levels in cell supernatant were measured by ELISA.TNF-α,IL-6,IL-1β,TLR4 and MyD88 mRNA levels were detected by RT-qPCR.Western blot was used to determine expressions of TLR4,MyD88,NF-κB,p-NF-κB,IκB-α and p-IκB-α proteins of cells.Results:Compared with control group,LPS-treated BV2 microglia had significantly lower viability(P<0.05),significantly higher NO secretion(P<0.05),significantly up-regulation levels of inflammatory factors TNF-α,IL-6 and IL-1β(P<0.05),and significantly higher mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),TLR4,MyD88,NF-κB,p-NF-κB,IκB-α,and p-IκB-α protein expressions were significantly increased(P<0.05).Compared with LPS group,BV2 microglia intervened with different concentrations of DG had significantly higher viability(P<0.05),significantly lower NO secretion(P<0.05),significantly lower TNF-α,IL-6 and IL-1β inflammatory factors levels(P<0.05),significantly lower mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),and TLR4,MyD88,p-NF-κB and p-IκB-α protein expressions were significantly reduced(P<0.05).Conclusion:DG can inhibit LPS-induced neuroinflammatory responses in microglia,whose underlying mechanism is suppression of TLR4/NF-κB signaling pathway.