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This study aimed to obtain a lentinan-specific fluorescent probe for the in situ visualization of lentinan. A carbohydrate-binding module with potential lentinan-binding capacity was discovered in a β-1,3-D-glucanase by using the bioinformatics techniques, and further recombinantly expressed. The binding specificity of the expressed protein, which was named as LBM was determined by the microtiter plate assays. The results indicated that LBM displayed a desired specificity for lentinan. The protein exhibited positive binding signals to lentinan, while could not bind to several examined polysaccharides including barley β-glucan, curdlan, or konjac glucomannan. Furthermore, the first lentinan-specific fluorescent probe was successfully constructed by fusing LBM with a green fluorescent protein EmGFP, based on which the in situ visualization of lentinan in Lentinula edodes was realized. It was shown that lentinan was not only presented in the mycelial cell wall, but also amorphously distributed in the interstitial space and inside the cell. The construction of the EmGFP-LBM provided a promising tool for the in situ visualization of lentinan.

Extremely low environmental temperature would lead the intracellular as well as the extracellular ice formation, decrease the fluidity and permeability of microbial cell membrane, cause abnormal folding of some proteins and nucleic acids in the cells and thus decrease the survival of the bacterial cells. In this paper, the effects of cold stress on membrane fluidity, protein and nucleic acid function of lactic acid bacteria (LAB) are reviewed, and the strategies adopted by LAB to overcome the multiple challenges encountered in cold stress are discussed, and the methodology employed to explore the cold stress on bacteria is also summarized. Perspective research of cold resistance of LAB should highlight on genomics, targeting on the critical genes involved in the synthesis of fatty acids, to unravel the mechanism of LAB in regulating membrane lipid composition. In addition, emphasis should be focused on the flexibility of the structure of chaperones, especially those of cold shock proteins on the resistance to the producer cells under cold stress.

Malware detection and homology analysis has been the hotspot of malware analysis.API call graph of malware can represent the behavior of it.Because of the subgraph isomorphism algorithm has high complexity,the analysis of malware based on the graph structure with low efficiency.Therefore,this studies a homology analysis method of API graph of malware that use convolutional neural network.By selecting the key nodes,and construct neighborhood receptive field,the convolution neural network can handle graph structure data.Experimental results on 8 real-world malware family,shows that the accuracy rate of homology malware analysis achieves 93%,and the accuracy rate of the detection of malicious code to 96%.

Background: Nasopharyngeal bacterial flora can cause respiratory tract diseases as well as invasive bacterial diseases. Moraxella catarrhalis colonizing in the nasopharynx is considered an important potential pathogen with an increasing production of β-lactamase. This study examined the nasopharyngeal colonization rate of M. catarrhalis and the antibiotic susceptibility of M. catarrhalis. Materials and Methods: Healthy children who visited one of the three University hospitals in the Republic of Korea or attended a day-care center around the participating hospitals were enrolled in this study. The nasopharyngeal samples were obtained by nasopharyngeal washing with normal saline and M. catarrhalis was isolated. The nasopharyngeal colonization rate of M. catarrhalis was investigated and the minimal inhibitory concentrations (MICs) were measured for commonly used oral antibiotics (amoxicillin, amoxicillin/clavulanate, cefaclor, cefixime, cefdinir, cefditoren, erythromycin and trimethoprim). Results: Three hundred and seventy-nine children aged between 6 months and years were enrolled, and the nasopharyngeal colonization rate of M. catarrhalis wa 33% (124 children). All isolated M. catarrhalis produced/β-lactamase. The MIC90 the antibiotics were as follows: amoxicillin, >16 mg/L; amoxicillin/clavulanate, 0.5 mg/L; cefaclor, 8 mg/L; cefixime, 0.125 mg/L; cefdinir, 0.25 mg/L; cefditoren, 0.25 mg/L; erythromycin, 0.5 mg/L; and trimethoprim, >16 mg/L. Conclusions: M. catarrhalis was colonized in 33% of the children aged 6 months to 5 years, and showed low MICs for amoxicillin/clavulanate and oral 2nd and 3rd generation cephalosporins. [ABSTRACT FROM AUTHOR]

Palliative care refers to the prevention and relief of physical and mental suffering through early recognition,active assessment,and management of pain and other painful symptoms to improve quality of life for both the patients with severe diseases and their families.A successful case of palliative care requires not only the establishment of correct concepts but also the team work and the improvement of the medical system.This paper introduced the end-of-life care experience for a patient with advanced lung cancer,showing the gains and deficiencies in the practice of palliative care.

SIRT6, a member of the silencing information regulatory protein family, is a nicotinamide adenine dinucleotide-dependent histone deacetylase and an ADP-ribose transferase enzyme. It plays an important role in fundamental physiological and pathological processes, including lipid metabolism, inflammation, oxidative stress and fibrosis, and is considered as a potential therapeutic target for metabolic syndrome. SIRT6 knockout mice displayed severe fatty liver, and the expression of SIRT6 in the liver of nonalcoholic steatohepatitis (NASH) mice was significantly lower than that of normal mice. Overexpression of SIRT6 significantly ameliorated NASH-induced liver damage. It is suggested that SIRT6 may play a key role in protecting against NASH. In this paper, we review the important regulatory functions of SIRT6 in the occurrence and development of NASH.

Geo-informatic spectrum analysis method was used to understand the complex geogra-phical phenomena concisely in graphic language. It is important for the integration research on spatial pattern and temporal process of land use change over multiple temporal and space scales. Based on remote sensing images in 1975, 1990, 1995, 2000, 2005, 2010 and 2015, we built the geo-spectrum of land use/cover change (LUCC) and quantitatively analyzed both the process and rend of LUCC in Manas River Basin. Results showed that the range of land use change in this basin was gradually decreasing, and land use structure tended to be simplistic, then tended to stable in later stage during 1975-1990. At the same period, the area of grassland increased greatly, which was mainly derived from unused land. The area of cultivated land expanded from 1990 to 2015, which mainly converted from unused land, forestland, and grassland. The geo-spectrum of land use change model showed that prophase change type, anaphase change type and continuous change type accounted for 1.3% of the total basin area. The overall characteristics of geo-spectrum unit were dominated by unused land converted to cultivated land, forestland and grassland. The land use change process was relatively stable. The Manas River Basin was covered 61.2% by region with comparatively stable spatial pattern. The most frequent way of land use change was increasingly expanding of cultivated land, which was displayed as: Liumaowan Bay Town of Shawan County and Liuhudi Town of Manas County were the center, then expanded to the two sides and spread to the Gurbantunggut Desert. The construct geo-spectrum of LUCC enriched the LUCC spatial-temporal analysis method system and provided an effective approach for the future research of LUCC under multiple temporal and spatial scales.

Snow samples were collected from the snow event on January 20-22, 2016 from 11 sites in downtown Hangzhou to explore the occurrence of sixteen perfluorinated compounds (PFCs) in the atmosphere. All samples were prepared by solid-phase extraction with Oasis WAX cartridges and analyzed using ultra performance liquid chromatography interfaced with tandem mass spectrometry (UPLC-MS/MS). The results showed that seven medium- and short-chain PFCs including C 4 and C 8 perfluorinated sulfonates (PFSAs) and C 4 -C 6 , C 8 , and C 9 perfluorinated carboxylic acids (PFCAs) were detected in the snow samples. Total PFC concentrations ranged from 2.85 to 35.1 ng·L -1 , whereas perfluorooctanoic acid (PFOA) dominated, with ranges of 2.15-23.0 ng·L -1 , and perfluorooctane sulfonate (PFOS) was detected at lower levels, ranging from 0 to 0.46 ng·L -1 . As compared to the results from other studies, the PFOA concentrations of the study region were at mid-level and the PFOS concentrations were relatively low. The spatial distribution of PFCs varied, and the pollutant concentrations of the snow samples from the sampling sites located in the urban areas were higher than those in the rural areas. The greatest total PFC concentrations were detected in Fuyang, whereas the lowest concentrations were detected in Jiande and Chun'an. In this study, the high concentrations of PFCs dominated by PFOA that were measured in the Hangzhou snow samples emphasize the importance of atmospheric wet deposition as one of the sources of PFC contamination in this ecological system and should be addressed. The occurrence of PFCs in the air, indicated by their presence in the snow, suggests that the atmospheric environment may be an important contributor in human and ecological exposure to PFCs by local residents.

Objective: To evaluate the visual development and prevalence of amblyopia, strabismus among preschool children., Methods: A random sample survey was performed in 4 610 preschool children from both urban and rural, aged 3 to 6 years. Participants underwent eye examination including visual acuity, refractive status, eye position, strabismus and amblyopia., Results: Percentage of visual acuity above 1.0 was 28.4%, 39.3%, 46.2% and 76.5% in children of 3, 4, 5 and 6-year-old group, respectively. The mean visual acuity of each group was 0.63 ± 0.19 in 3-years old, 0.69 ± 0.16 in 4-year-old, 0.71 ± 0.22 in 5-year-old, 0.79 ± 0.29 in 6-year-old. Rural children have better vision acuity compared with those from urban. Hypermetropic was frequent refractive errors. Manifeststrabismus was found in 2.21%, with exotropia being more prevalence than esotropia; detection rate of recessive strabismus was 33.52%, mainly being exophoria; Based on current diagnostic criteria, the prevalence of amblyopia were 2.93% in 6 year-old group, 4.81% in 5-year-old group, 16.21% in 4-year-old group, 33.33% in 3-year-old group., Conclusion: Vision acuity is increasing with age in preschool population. A diagnosis standard of amblyopia suitable for each age group should be established to substitute the current one which has a high visual standard for amblyopia. Refractive error, strabismus and amblyopia are the leading causes of visual impairment among preschool-aged children, which represent the focus of prevention of blindness in preschool children.

Objective: To investigate the cellular mechanisms of adipocyte lipolytic response to a high concentration of glucose, which liberates free fatty acids (FFA) efflux from adipose cells to plasma., Methods: Adipocytes were isolated from epididymal fat pads of Sprague-Dawley (SD) rats, and were incubated in the presence or absence of excess glucose (25 mmol/L). Glycerol released in the culture medium was determined by use of a colorimetric assay and served as an index of lipolysis. The data are expressed as micromoles glycerol per liter packed cell volume (PCV) of adipocytes. The protein levels of native and phosphorylated perilipin, hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) were examined by immunoblotting. Adipose lipases activity was determined by performing an enzymatic assay., Results: A high concentration of glucose at 25 mmol/L caused a significant increase in lipolysis in primary rat adipocytes. A 24 h incubation with 25 mmol/L glucose elevated glycerol accumulation from 4.4 mmol/L PCV to 6.4 mmol/L PCV, by 1.5 fold (P<0.01) and promoted glycerol release from 0.11 mmol/L PCV to 0.24 mmol/L PCV, by 2.1 fold (P<0.01). The lipolytic response mediated by high glucose occurred apparently at the end of 16 h and remained high after 24 h incubation. The lipolytic action of glucose at 25 mmol/L was more effective than that at 10 mmol/L. To elucidate the mechanism by which high glucose stimulated lipolysis, we showed that high glucose promoted the phosphorylation of perilipin but did not alter its protein level. Further, we showed that high glucose caused a significant increase in adipose lipases activity by 1.74 fold. The protein level of HSL but not ATGL was significantly upregulated by high glucose. Moreover, the acute lipolytic response to isoproterenol was further enhanced by high glucose., Conclusion: High glucose directly stimulates the releases of glycerol and FFA from adipocytes; the cellular mechanisms of this lipolytic action involve promoted perilipin phosphorylation, augmented HSL expression and increased adipose lipases activity under high glucose conditions. These results suggest that a high concentration of glucose alone can directly liberate FFA from adipocyte to plasma, hence increasing systemic FFA levels and impairing insulin sensitivity.

Objective: To describe a community-based model for prevention and control of severe alpha and beta thalassemias in Zhuhai city of Guangdong province., Methods: Couples for premarital medical examination or regular healthcare examination in pregnancy were enrolled in this prospective screening program, which was supported by the two-level network composed of 6 local hospitals for testing thalassemias and follow-up for genetic counseling. A conventional heterozygote screening strategy was used to determine alpha and beta thalassemia traits in women and their partners according to the standard procedures of hematological phenotype analysis. Then confirmative diagnosis of alpha and beta thalassemia was performed on those couples suspected at-risk for severe thalassemia by using the PCR-based molecular diagnostic assays. The couples at-risk for severe thalassemia were counseled and offered prenatal diagnosis and termination of pregnancy in case of an affected fetus., Results: During the period between January 1998 and December 2005, the screened records included 85522 young females and their partners for premarital screening and 10439 pregnant women for prenatal screening, with 71.38% coverage of total population recorded in this city for premarital screening. Six thousands five hundreds and sixty-three individuals in total were found to be the carriers of thalassemias, with 4312 for alpha thalassemia (4.5%) and 2251 for beta thalassemia (2.3%), respectively. One hundred and forty-eight couples were diagnosed to be at-risk for thalassemias, including 103 for alpha thalassemia and 45 for beta thalassemia, respectively. Successful prenatal diagnosis was made for 142 (98 for alpha thalassemia and 44 for beta thalassemia) out of 148 (95.9%) pregnancies at-risk for severe thalassemias. Twenty-three cases of hydrops fetalis, 4 of Hb H diseases and 14 of beta thalassemia were identified. All 41 pregnancies with affected fetuses were voluntarily terminated. Thus, this has led to a marked decrease of severe thalassemia syndrome since the program started., Conclusion: We presented the first community-based prospective screening program in China for control of alpha and beta thalassemia in Zhuhai city with a population of 1.29 million through premarital or prenatal screening. This model could be used for control of thalassemias and other hemoglobinopathies in other regions of China and also in other developing countries.

This study was aimed to investigate the effects of DMSO on platelets during pre-treatment for lyophilization, including centrifugation, washing and loading trehalose. After pre-treatment for lyophilization, the expression of platelet membrane surface glycoprotein (GP) including CD62p and PAC-1 was analyzed by FCM before and after induction with thrombin, the mean platelet volume (MPV) and platelet maximal aggregation with several platelet inducers were investigated. The results showed that the expression rates of CD62p and PAC-1, as the platelet activation signs, increased and were 30.37% and 15.01% respectively in group without DMSO after pre-treatment. And their differences in comparison with control were statistically significant, but that of CD62p was 10.72% and PAC was 10.11% in group with DMSO, in comparison with group without DMSO respectively, their differences were statistically significant after diluting with DMSO, CD62p was re-expressed to 54.39% in group with DMSO and more than that in group without DMSO and lower than control statistically significant. PAC-1 was re-expressed to 49.28% in group with DMSO and more than that in group without DMSO (p<0.01) and reached to control. Platelet maximal aggregations induced by thrombin, restocetin and propyl gallate were 92.76%, 91.24% and 89.66 respectively in group with DMSO. These were closed to that in control group and in group without DMSO. But the aggregation induced by ADP was 34.33%, it was less than control (p<0.01) and more than that in group without DMSO (p<0.01). It is concluded that DMSO can inhibit the expression of CD62p and PAC-1 on platelet in vitro. But when diluted with plasma, platelets can express CD62p and PAC-1 induced by thrombin and be led to aggregate by several inducers, so the inhibitory effects of DMSO on platelet activation are reversible. DMSO play roles in inhibitor damage from platelet activation and cryoprotectant. This property of DMSO is very important in research of platelets lyophilization.

This study was aimed to compare the difference of quality between buffy-coat-collected platelet concentrates (BC-PC) and single-donor plateletpheresis (SDP). 15 packs of BC-PC and 15 units SDP were stored at 20 degrees C - 24 degrees C with agitation. Platelet concentration, platelet volume, residual leukocyte and residual erythrocyte in two groups were examined after preparation for 1 hour. Mean platelet volume, pH value, hypotonic shock response (HSR), CD62p expression and CD62p re-expression of platelet were detected on 0, 1, 2, 3, 4, 5 days of platelet preservation. The results showed that the platelet yields, residual leukocyte and residual erythrocyte in two groups accorded with the national quality standard respectively, but residual leukocyte and residual erythrocyte in BC-PC group were higher than those in SDP group when platelet yields in two groups were equal (p < 0.01). Lactate concentration, CD62p expression of platelet increased with prolongation of preseved time, while pH value decreased gradually. Compared with SDP group, there were significant differences in CD62p expression, CD62p re-expression of platelet preserved for 0 - 5 days (p < 0.01), and in pH value of platelet preserved 2 - 5 days (p < 0.01). There was no changes in HSR of SDP group for 0 - 5 days, while HSR in BC-PC group decreased gradually. There were significant differences in HSR of platelet preserved for 1 - 5 days (p < 0.01). It is concluded that the platelet concentrates prepared by BC-PC are not equal to SDP in quality, the preparation technology of BC-PC should be optimized further in order to reduce residual leukocyte, residual erythrocyte and activated platelet yields, as well as improve the quality of BC-PC.

Objective: To assess the value of pulsed Doppler tissue imaging (PW-DTI) in evaluating the left ventricular diastolic dysfunction (LVDD) in patients with type 2 diabetes mellitus (DM)., Methods: Seventy-seven type 2 diabetic patients and 43 healthy volunteers underwent PW-DTI and mitral inflow pulsed wave Doppler (MPWD). Six positions of the mitral annulus were measured by PW-DTI for early diastolic velocity (Em), end diastolic velocity (Am), Em/Am ratio. Early mitral inflow velocity (E), end mitral inflow velocity (A), left ventricular isovolumetric relaxation (IVRT) and E/A ratio were measured by MPWD., Results: In the diabetic patients, Em, Em/Am and E/A ratios were significantly decreased and Am and A increased with prolonged IVRT. The two examination modalities showed a significant correlation between Em/Am and E/A. The detection rate of LVDD in the diabetic group was 94.81% by PW-DTI and 79.22% by MPWD., Conclusion: PW-DTI and MPWD show a significant correlation in assessment of the left ventricular filling, but the former displays better performance in sensitive and accurate diastolic function evaluation and provides early evidence for of diabetic cardiomyopathy of due to its potent ability in differentiation of false-normal presentations.

Perilipins are the proteins associating with the lipid droplets in adipocytes and steroidogenic cells. Unphosphorylated perilipins coat the surface of intracellular lipid droplets to form a barrier that prevents lipase from accessing to triacylglycerol core, thus suppressing lipolysis. Upon activation of protein kinase A (PKA), two proteins, hormone-sensitive lipase (HSL) and perilipins, are phosphorylated. The phosphorylated perilipin is required for inducing the translocation of HSL from the cytosol to the lipid droplets of adipocytes and is essential for the initiation of lipolytic reaction. It is proposed that phosphorylation of perilipin is a key step for the activation of lipolytic cascade via PKA and ERK signaling pathways. Dysregulation of perilipin involves in the pathogenesis of obesity, diabetes and atherosclerosis.

Objective: To screen efficient siRNA for inhibiting hepatitis B virus using the technique of PCR-based tRNA(val) Pol III-shRNA expression cassettes (SECs)., Methods: Based on core gene sequence of HBV, five target sites of siRNA were designed. tRNAval Pol III-shRNA expression cassettes produced by one-step overlapping extension PCR strategy were co-transfected with HBV C gene and pC-EGFP plasmid into AD293 cells respectively. Forty-eight hours after transfection, fluorescence of HBVC-GFP protein was detected by fluorescence-activated cell sorting (FACS); HBV C mRNA was detected by semi-quantitative RT-PCR. HBV-producing HepG2. 2. 15 cells were transfected with selected SECs for 72 h, HBsAg and HBeAg in the cell culture medium were detected by radioimmunoassay assay (RIA). HBV pgRNA from cell total RNA was detected by semi-quantitative PCR., Result: Co-transfection with pC-GFP plasmid and SECs into AD293 cells resulted in inhibition expression of HBV C gene and decrease of EGFP fluorescence intensity. SEC-492i showed most significant inhibition effect on HBV C-EGFP expression compared with other SECs. Selected SEC-492i or SEC-282i targeting core gene could efficiently decrease expression of HBeAg and the level of HBV pgRNA in a dose-dependent manner. SEC-492i inhibited HBV replication and antigen expression in a more efficient way than SEC-282i at the same final concentration., Conclusion: The expressed shRNA, which targets sites on HBV C mRNA in 492i, is to have having most efficient RNAi effect. tRNAval Pol III-shRNA expression cassettes produced by one-step overlapping extension PCR strategy should be useful for identification of optimal siRNA.

Objective: To observe the protective effect of diammonium glycyrrhizinate on hepatic function in burn patients., Methods: Twenty burn patients with hepatic dysfunction were enrolled in the study and were randomly divided into 2 groups, i. e. treatment (T, n = 10, with conventional treatment and intravenous infusion of 150 mg diammonium glycyrrhizinate per day for 14 days), and control (C, n = 10, with conventional treatment) groups. The blood samples in both groups were collected before and 7 and 15 days after the treatment. The serum contents of ALT, AST, GGT, ALP and PA in the blood samples were determined and analyzed comparatively., Results: There was obvious difference in the serum contents of ALT, AST, GGT, ALP and PA in the T group before treatment (168 +/- 46 U/L, 104 +/- 29 U/L, 162 +/- 37 U/L, 149 +/- 17 U/L, 310 +/- 35 mg/L, respectively) and 15 days after treatment (51 +/- 9 U/L, 31 +/- 3 U/L, 56 +/- 10 U/L, 103 +/- 9 U/L, 372 +/- 44 mg/L, respectively, P < 0.05). There was no difference in these indices in the C group before and after treatment (P > 0.05)., Conclusion: Diammonium glycyrrhizinate seemed to be beneficial to the management of postburn hepatic dysfunction with obvious rapid depression of hepatic enzymes.

Objective: To develop an effective report gene system to test the effect of small interfering RNA (siRNA)., Methods: HBV S gene was fused with enhanced green fluorescent protein (EGFP) gene to form HBs-GFP and the plasmid containing HBs-GFP was constructed. A vector expressing small hairpin RNA (shRNA) pAVU6 + 4sh357 was also constructed. Two plasmids were co-transfected into HepG2 cells transiently. The fluorescence of HBs-GFP was detected by fluorescence-activated cell sorting (FACS). The mRNA expression in HepG2 cells was detected by conventional RT-PCR and real-time PCR., Results: siRNA inhibited the expression of HBs-GFP 72 hours post transfection. The fluorescence of HBs-GFP in HepG2 cells treated with pAVU6+4sh357 was reduced by 55.4% compared with that of controls. The HBs-GFP expression in HepG2 cells treated with pAVU6+4sh357 was reduced by 76.3% and 90% as measured with conventional RT-PCR and real-time PCR, respectively., Conclusion: This investigation demonstrated siRNA derived from shRNA expression vectors can inhibit the expression of HBs-GFP in HepG2 cells.

Objective: To find some effective short interfering RNA's sites targeting HBV surface gene sequence using shRNA expression vectors., Methods: Four shRNA expression vectors targeting HBV surface gene sequence were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with HBs-EGFP fusion gene plasmid. The changes of HBs-EGFP image were detected by FACS and microscopy. The HBs-EGFP mRNA expression was evaluated by RT-PCR., Results: Four shRNA expression vectors and HBs-EGFP fusion gene plasmid were successfully constructed. pAVU6 + 4sh579 vector inhibited the HBs-EGFP expression by 69.8% in AD293 and suppressed the HBs-EGFP mRNA expression by 74.6%., Conclusions: The results showed that the 579 site of HBV surface gene sequence was an effective target and pAVU6 + 4sh579 vector could suppress the HBs-EGFP expression in AD293 cells