Objective To evaluate the effect of linoleic acid on lipopolysaccharide (LPS)-induced release of inflammatory factors in the macrophages of mice.Methods The peritoneal macrophages obtained from C57BL/C mice were seeded in 24-well plates at a density of 4× 105 cells/well and in 6-well plates at a density of 2× 106cells/well.The cells were incubated and attached to the wall overnight in a 5％ CO2 incubator in humidity at 37 ℃.The experiment was performed in 2 parts.Part Ⅰ The cells in 24-well plates were randomly divided into 5 groups (n =8 each) using a random number table:control group (group C);LPS group;3 different concentrations of linoleic acid groups (LA1-3 groups).The sterile anhydrous alcohol 1 μl was added in group LPS,0.1,0.5 and 1.0 mol/ml linoleic acid 1 μl were added in LA1-3 groups,respectively,and 30 min later 100 μg/ml LPS 1 μ,l was added in LPS and LA1-3 groups.The culture medium was collected at 6 h after LPS administration to measure the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the supernatant by enzyme-linked immunosorbent assay.PartⅡ The cells in 6-well plates were randomly divided into 3 groups (n =6 each) using a random number table:control group (group C);LPS group;0.5 mol/ml linoleic acid group (group LA).The sterile anhydrous alcohol 1 μl was added in group LPS,0.5 mol/ml linoleic acid 1 μl was added in group LA,and 30 min later 100 μg/ml LPS 1 μl was added in LPS and LA groups.At 1 h after administration of LPS,the expression of Toll-like receptor 4 (TLR4) was determined by flow cytometry,and the expression of phosphorylated nuclear factor kappa B (NF-κB) p65 (p-NF-κB p65),phosphorylated extracellular signal-regulated protein kinase (p-ERK) and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) was determined by Western blot.Results Part Ⅰ Compared with group C,TNF-α and IL-6 concentrations in the supernatant were significantly increased in LPS and LA1 3 groups (P ＜ 0.05).Compared with group LPS,TNF-α and IL-6 concentrations in the supernatant were significantly decreased in LA1 3 groups (P＜0.05).Compared with group LA1,TNF-α and IL-6 concentrations in the supernatant were significantly decreased in LA2 and LA3 groups (P＜0.05).Compared with group LA2,TNF-α and IL-6 concentrations in the supernatant were significantly decreased in group LA3 (P ＜ 0.05).Part Ⅱ Compared with group C,the expression of TLR4,p-NF-κB p65,p-ERK and p-p38 MAPK in macrophages was significantly up-regulated in LPS and LA groups (P＜0.05).Compared with group LPS,the expression of TLR4,p-NF-κB p65,p-ERK and p-p38 MAPK in macrophages was significantly down-regulated in group LA (P＜0.05).Conclusion Linoleic acid can inhibit LPS-induced release of inflammatory factors in the macrophages of mice,and the mechanism may be related to the inhibition of TLR4 signaling pathway activation.

Objective To investigate the feasibility of macrophages as cell carriers to deliver floate modified oxygen loaded ultrasound contrast agent . Methods The phagocytic activity of macrophages was analyzed by ink phagocytose test , and the expression of folate recepters ( FRs ) on macrophages cell membrane surface was tested by immunofluofluorescence assay . Oxygen/paclitaxel loaded lipid microbubbles( OPLMB) and folate‐targeted OPLMB ( TOPLMB) were synthesized by mechanical shock method and incubated with macrophages in vitro . According to different treatment conditions ,the cells were divided into three groups:group A ( OPLMB) ,group B ( free folic acid + TOPLMB) and group C ( TOPLMB) , the fluorescence intensity of the cells were observed under fluorescence microscope ,and the phagocytic percentage and the phagocytic index of macrophages uptake OPLMB and TOPLMB were observed by bright field microscope . Results The phagocytic percentage of macrophages phagocytose ink was (99 .3 ± 1 .0)% ,FRs was highly expressed on macrophages cultured in vitro . After incubation for 30 minutes ,the fluorescence intensity of group C was significantly higher than those of A and B ,the phagocytic percentage in three groups were (19 .5 ± 0 .2)% ,(21 .0 ± 0 .2)% and (81 .2 ± 10 .0)% respetively . The phagocytic percentage of group C were significantly higher than those in group A and group B ( P <0 .05) . Conclusions Macrophages cultured in vitro possess highly phagocytic activity and these cells highly express FRs ,and can be used as cell carriers to deliver floate modified oxygen loaded multimodality ultrasound contrast agent .

ObjectiveTo analyze the clinicopathologic features of hereditary cutaneomucosal venous malformation (VMCM) in a Chinese family.MethodsFamily history was investigated in a family with VMCM,and tissue specimens were obtained from the lesions of the proband and subjected to histopathological analysis.ResultsAmong 65 members from 5 generations of the family,19 were affected by VMCM,hinting an autosomal dominant inheritance.None of the family members experienced gastrointestinal bleeding,central nervous system disorders,or cardiac defects.Affected individuals usually presented with multiple irregularly sized,blue-violet,elevated and slightly indurated masses located in the oral mucosa and subcutaneous tissue of the extremities.Pathological analysis showed malformed veins with abnormally dilated cavities and irregularly thickened walls.Although small veins were abnormally proliferating and clustered,there was no endothelial discontinuity.The smooth muscle layer was thickned in a varying degree or absent.ConclusionA diagnosis of VMCM is made according to the inheritance manner,clinical manifestation and pathological findings.

Objective To evaluate the knowledge,attitude and practice(KAP)related with AIDS of local floating population.Methods A total of 1 942 floating persons were sampled by multistage cluster sampling and interviewed with questionnaire in the agricultural markets,construction sites,and their habitats within 3 communities in Hongkou District.Results 82.9% of the interviewees were 20 ～49 years old.The average score was 34.77 ±3.52 (maximum score is 44)for knowledge,12.11 ±2.32(maximum score is 19)for attitude,5.50 ±0.95(maximum score is 7)for practice,respectively.The scores increased with educational level significantly.The scores of knowledge were 33.26 ±3.54,34.63 ±3.23,36.56 ±3.39 among the subjects with educational levels of primary school and below,junior high school,senior high school and above.The scores of attitude were 13.77 ±2.27,14.79 ±2.39,15.62 ±2.38,respectively.And the score of practice was 5.40 ±0.90,5.51 ±0.93,5.58 ±1.03,respectively.Conclusion In the present,the KAP relating AIDS of local floating population is poor in Hongkou District.A variety of intervention methods and operational patterns should be developed for the floating population with different educational levels and jobs.

Objective: To prepare a nanoprobe, anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods: The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDS-PAGE. The GD/ScFvMEL-QDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC-803 cells and melanoma A375 cells. Results: The recombinant pET32a-GD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDS-PAGE. The purified- and refolded-GD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMEL-QDs nanoprobe was successfully prepared. The GD/ScFvMEL-QDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC-803 cells. Conclusion: We have successfully prepared an anti-human melanoma ganglioside single-chain antibody-CdTe quantum dot nanoprobe, which can specifically bind melanoma cells.

Objective To examine the prevalence of depressive symptoms and related factors among HIV-positive men who have sex with men (MSM) in Shanghai.Methods All of the HIV-positive MSM were enrolled to participate in a cross-sectional study with questionnaire-based interview during Nov.,2014 to Nov.,2015.Depression was measured with a 12-item version of the Center for Epidemiological Studies Depression Scale (CES-D),where a score of 9 had been recommended as the cutoff score to indicate possible depressive symptoms.Results A total of 505 participants were recruited with a median age of 30.Among them,the majority were aged 21-44 (77.2％),non-local resident (64.6％),unmarried (73.1 ％) and university graduate or above (62.1 ％).Depressive symptoms were detected from 235 MSM and the prevalence of depression was 46.5 ％.Multiple logistic regression analysis showed that MSM who were non-local residents (OR =1.66,95 ％ CI:1.01-2.49)and had more than one male sex partner in the past year (OR=2.21,95％ CI:1.43-3.41) were more likely to have depressive symptoms compared with local residents and had only one male sex partner or less.On the contrary,compared with the education level of junior middle school or below and having no long-term partners,those who were senior high school educated (OR=0.39,95％ CI:0.21-0.74) and had homosexual behavior with main long-term partners without condoms (OR =0.38,95 ％ CI:0.17-0.85) had lower risk to be depressive.Conelusions The prevalence of depressive symptoms among HIV-infected MSM is high.Residence,education level,homosexual behavior with main long-term partner and number of male sex partner are significantly associated with depressive symptoms,suggesting psychological interventions should be further explored and studied.

The fermentative H2-producing strain Clostridium sp. H-61 was isolated from anaerobic sludge,was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production ability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production ability. which was measured in the batch culture experiments. With the condition of 10 g/L glucose,its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L and 33.19 mmol H2/g DW?h,69.89% and 68.14% higher than that of the original strain,respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type,while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation studied,including time,carbon source,nitrogen source,glucose concentration,glucose utilization,initial pH and incubation temperature had been studied,indicated the optimum condition of hydrogen production for the mutantHCM-23 as initial pH 6.5,temperature 36 ℃,and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different,such as,the growth lag phase and the utilization of inorganic nitrogen source,etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

OBJECTIVETo understand the characteristics of HIV/AIDS patients with late diagnosis and find the factors associated with late HIV detection.

METHODSHIV late diagnosed patients and early diagnosed patients, which were identified and classified by definition in advance, were selected from the case reporting database of HIV/AIDS Comprehensive Response Information Management System in eight counties of four provinces (Zhumadian, Nanyang, and Zhoukou of Hennan province; Liuzhou and Lingshan county of Guangxi autonomous region; Guangzhou and Shenzhen of Guangdong province; Dehong of Yunnan province) between January 1, 2009 and June 30, 2010. A total of 3912 eligible patients were investigated, including 2496 late diagnosis and 1416 early diagnosis. The structured questionnaires were used to obtain information on behaviors, HIV detection history and reason of late detection for all eligible HIV/AIDS patients. Late diagnosed patients were defined by CD4 T-cell counts less than 200 cells/mm(3) or diagnosis as AIDS within the reported year after the first HIV positive test. The univariate and multivariate logistic regression methods were used to analyze the characteristics of HIV/AIDS late diagnosed patients.

RESULTSOnly 14.2% (350/2469) of them have ever had the awareness of "to go for HIV testing", 68.8% (150/218)of which did not put it into practice within one month because of discrimination and stigma. Among those HIV late diagnosed patients without the awareness of "to go for HIV testing", the proportions of "never worried about HIV infection" or "never heard of AIDS" were 69.7% (1476/2116) and 18.1% (383/2116), respectively. When those HIV late diagnosed patients visited health settings because of AIDS related symptoms, only 40.0% (590/1475) of them received the HIV testing service. Furthermore, 54.5% (322/590) of those received HIV testing were not informed the results. Compared with early diagnosed patients, patients with late diagnosis were over 50 years old (OR = 4.14, 95%CI: 3.09 - 5.55), primary school education (OR = 1.29, 95%CI: 1.10 - 1.52) and illiteracy (OR = 2.15, 95%CI: 1.25 - 2.82), Routes of transmission from former illegal blood or plasma (OR = 2.91, 95%CI: 2.27 - 3.74) and transfusion of blood/blood products (OR = 2.79, 95%CI: 2.11 - 3.68). Late diagnosed patients were identified mainly from voluntary counseling and testing (45.4%, 1130/1528) and medical institutions (38.3%, 954/1469).

CONCLUSIONThe main reasons for late diagnosis of HIV infection are low initiative of HIV testing and discrimination and stigma. Furthermore, the low awareness of medical institutions to actively provide HIV testing affects the early diagnosis of HIV infections.

Acquired Immunodeficiency Syndrome ; diagnosis ; epidemiology ; Adult ; China ; epidemiology ; Counseling ; Delayed Diagnosis ; Female ; HIV Infections ; diagnosis ; epidemiology ; Humans ; Male ; Mass Screening ; Middle Aged ; Risk Factors

OBJECTIVETo study the mechanism of Huogu I formula (I) in treating osteonecrosis of femoral head.

METHODSForty-eight healthy female Leghorn chickens were randomly divided into control group, model group and Huogu I group, and each group consisted of 16 chickens. At the meantime of model establishment, chickens of the Huogu I group were administrated with decoction, while the model and control group with distilled water by gavage. At the 8th and 16th week after medication, blood samples were obtained for blood lipid detection while both sides of femoral head were harvested for the rest of examinations. Specifically, expressions of bone morphogenetic protein-2 (BMP2), transforming growth factor beta1 (TGFβ(1)), Smad4 and Smad7 were evaluated by immunohistochemistry, while expression of osteoprotegerin/receptor activator of nuclear factor kappaB ligand (OPG/RANKL) mRNA was detected by in situ hybridization.

RESULTSCompared with the control group, serum levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in the model group rose significantly. Positive cell counting of BMP2, TGFβ(1), Smad4 and OPG in femoral head of the model group dropped prominently. Positive cell counting of Smad7 and RANKL increased dramatically. In contrast with the model group, levels of TC, TG and LDL-C in Huogu I group reduced significantly. Positive cell counting of BMP2, TGFβ(1), Smad4 and OPG in femoral head of the Huogu I group increased prominently. Indices of Smad7 and RANKL both decreased significantly. Especially at the 8th week, these variations were more significant.

CONCLUSIONHuogu I formula is effective in promoting repair of necrotic femoral head by regulating the expressions of BMP2, TGFβ(1), Smads and OPG/RANKL of osteoclast in femoral head.

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Bone Regeneration ; drug effects ; physiology ; Chickens ; Chondrocytes ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Femur Head Necrosis ; chemically induced ; drug therapy ; metabolism ; Lipid Metabolism ; physiology ; Osteocytes ; metabolism ; Osteoprotegerin ; genetics ; metabolism ; Receptor Activator of Nuclear Factor-kappa B ; genetics ; metabolism ; Smad4 Protein ; metabolism ; Smad7 Protein ; metabolism ; Steroids ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

In vitro neuraminidase inhibition assays and ultrafiltration liquid chromatography with diodearray detector coupled to time of flight mass spectrometer (UPLC-DAD-TOF-MS) were combined to screen bioactive compounds inhibiting neuraminidase from Isatidis Radix. By comparing the compounds from Isatidis Radix before and after ultrafiltration, we found that arginine, goitrin and adenosinea can bind with neuraminidase, and the binding degree of the three compounds were (36.23 +/- 1.12)%, (32.54 +/- 1.02)% and (9.38 +/- 0.47)%, respectively. The IC50 of arginine and goitrin were (1.16 +/- 0.02), (1.20 +/- 0.02) g x L(-1), respectively. While the IC50 of adenosinea was higher than 500 g x L(-1). The results showed that arginine and goitrin might be the main compounds with antiviral activity of Isatidis Radix. This study may provide a useful method for the screening of bioactive compounds and quality control of Isatidis Radix.
Antiviral Agents ; analysis ; pharmacology ; Arginine ; analysis ; pharmacology ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Isatis ; chemistry ; Mass Spectrometry ; Neuraminidase ; antagonists & inhibitors ; metabolism ; Orthomyxoviridae ; drug effects ; enzymology ; Oxazolidinones ; analysis ; pharmacology ; Plant Roots ; chemistry ; Ultrafiltration ; Viral Proteins ; antagonists & inhibitors ; metabolism