Zinc oxide nanoparticle (nano-ZnO) has a size between 1 and 100 nm. Nano-ZnO has some special effects, such as small size effect, surface effect, quantum size effect, which makes it different from the ordinary ZnO, and is widely used in rubber industry, food processing, cosmetics and pharmaceutical fields. It has been reported that nano-ZnO has toxic effects in vitro and in vivo, but the mechanism of toxicity is still unclear. Therefore, it is important to evaluate the safety nano-ZnO by studying its biological toxic effects and related mechanisms. In this paper, we summarize the characterization, ingestion pathway, metabolism, systematic toxicity of nano-ZnO and its mechanisms, which may provide us with new strategy for the toxic research of nano-ZnO.
Animals ; Humans ; Metal Nanoparticles ; toxicity ; Zinc Oxide ; toxicity

OBJECTIVETo investigate the epidemic characteristics of hand-foot-and-mouth disease (HFMD) in children and exposed population in Hangzhou city.

METHODSThe throat swab or stool samples from children with HFMD admitted in Hangzhou Children's Hospital were collected. The HFMD pathogens were detected by real-time fluorescent quantitative PCR. The distribution of different HFMD pathogens in HFMD patients was subsequently determined. Human enteric virus type-71 (HEV71) in stool samples from subjects, who had close or general contact to 54 severe HFMD children with positive HEV71, was detected, and these contact persons were followed-up for one month. The diversity of predominant pathogens of HFMD in the area during 2011-2013 was investigated.

RESULTSIn 641 HFMD children, the male/female ratio was 1.4:1 and 80.3% was 1-3 years old. HEV71 was detected in 24.3% HFMD children (156/641), while coxsackievirus group-A type-16 (CVA16) and other enteroviruses were detected in 4.7% (30/641) and 71.0% (455/641) of the cases, respectively. 75.6% (118/156) of HEV71-infected cases were diagnosed as severe HFMD cases, while those for CVA16-infected and other HFMD viruses-infected were 13.3% (4/30) and 6.2% (28/455) respectively (Χ(2)=43.28, P<0.05). HEV71 was the predominant HFMD pathogens during 2011-2012, while the predominant HFMD pathogens in 2013 were the other HFMD viruses. In the 54 close contact persons or 54 general contact persons, 9 or 10 persons were detectable for HEV71, but no clinical symptoms of HFMD were presented.

CONCLUSIONThere are no marked changes of epidemic seasons, favorable age and gender ratio of HFMD in Hangzhou area in 2013. The infection of HEV71 tends to cause the severe HFMD but the other enteroviruses have substituted HEV71 as the predominant pathogens of HFMD.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Infant ; Male

OBJECTIVETo evaluate the quality of life in patients with chronic obstructive pulmonary disease (COPD) by COPD Assessment Test (CAT) and Body Mass Index, Airflow Obstruction, Dyspnea, Exercise Capacity Index (BODE).

METHODSOne hundred patients with stable COPD admitted in Putuo People's Hospital were recruited in the study. CAT and BODE index were measured for each patient.The deaths and frequency of exacerbations were recorded during 3-year follow-up period,and the correlation between CAT and BODE in evaluating COPD prognosis was analyzed.

RESULTSThere were 28, 30, 29 and 13 patients with CAT score of 1, 2, 3 and 4, respectively; while there were 31, 29, 28 and 12 cases with BODE scores of 1, 2, 3 and 4. CAT scores were well correlated with BODE evaluation in terms of overall score and scores of 4 items (r= -0.237, -0.772, 0.789, -0.767, 0.888, respectively, Ps<0.05). COPD exacerbation incidence and mortality increased with the increasing CAT levels. The rank sum test showed that there were no significant differences between CAT and BODE index in the frequency of acute exacerbation(P<0.05); and in the death toll, the difference was not significant(1 group Χ2=0.919, 2 group Χ2=0.001, 3 group Χ2=0.177,4 group Χ2=0.322, Ps>0.05).

CONCLUSIONCAT is relevant to BODE in evaluating incidence of exacerbation and mortality for patients with COPD and CAT is more easily to be applied.

Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Pulmonary Disease, Chronic Obstructive ; diagnosis ; Quality of Life

OBJECTIVETo investigate the mechanism of Mycobacterium tuberculosis invasion to mouse dendritic cells (DC).

METHODSMycobacterium tuberculosis strain H37Rv was co-cultured with mouse DC2.4 cells.The mRNA expression of Toll-like receptor 2/4(TLR2/4) in DC2.4 cells was detected by fluorescent quantitative real-time PCR and the protein expression of nuclear factor κB(NF-κB) was assessed by Western blotting.The extracellular concentration of tumor necrosis factor α(TNF-α) was measured by ELISA methods during Mycobacterium Tuberculosis invasion.Indirect immunofluorescent staining and flow cytometry assay were used to detect the expression of CD80 and CD86 on DC2.4 cells before and after invasion.

RESULTSThe invasion of Mycobacterium tuberculosis in DC2.4 cells was observed after 2 h of co-incubation.The rates of invasion were (37.9±5.6)%,(51.2±7.6)%,(57.2±8.9)% and(63.9±6.8)% at 6,8,10 and 12 h after co-incubation,respectively.The mRNA expression level of TLR2 /4 was significantly increased at 6 h but decreased at 10 h after co-incubation.The expressions of NF-κB p65 and TNF-α were higher in DC2.4 cells after being invaded by 6,8,and 10 h and then gradually decreased.CD80 and CD86 expression were increased on DC2.4 at 6 h after co-incubation.

CONCLUSIONInvasion of Mycobacterium tuberculosis strain H37Rv to DC might enhance its antigen-presenting function through activation of TLR2/4-NF-kB signaling pathway.

Animals ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Cells, Cultured ; Dendritic Cells ; immunology ; metabolism ; Mice ; Mycobacterium tuberculosis ; NF-kappa B ; metabolism ; Signal Transduction ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo construct and identify lentiviral vector containing human ILK-shRNA and mda7 gene.

METHODSBased on the human ILK gene sequences, RNAi target sequences were designed and cloned into the lentiviral vector pSicoR-eGFP by restriction endonuclease HpaI and XhoI double digestion and T4 DNA ligase ligation. Based on the human mda7 gene sequences, PCR primers were designed to clone the full-length mda7, and were cloned into the lentiviral vector pLVX-Puro. After the candidate clones were identified by DNA sequencing, the recombinant plasmid and the three packaging plasmids were co-transfected into the human embryonic kidney 293T cells by lipofectamine 2000 to produce the lentiviral particles. Human prostate cancer PC-3 cells were infected with the constructed lentiviral vector. The ILK and mda7 expression levels in PC-3 cells were quantified by qPCR and Western blot, respectively. The effect of ILK and mda7 on proliferation and migration of PC-3 cells were assessed by MTT method and Transwell assay, respectively.

RESULTSILK-pSicoR-eGFP and mda7-pLVX-Puro lentiviral vectors were successfully constructed. Strong green fluorescence was observed in the 293T cells under the fluorescent microscope after co-transfection of 293T cells with 4 plasmids of lentiviral vector. The transfection efficiency of the collected virus exceeded 90% in the 293T cells and the PC-3 cells were infected with the lentiviral particles with high efficiency. The A and B lentiviral vector inhibited the expression of ILK at both the mRNA and protein levels in PC-3 cells significantly. The mda7-pLVX-Puro lentiviral vector increased the expression of mda7 in PC-3 cells, and the ability was maintained for one month. Within 96 h, ILK and mad7 significantly inhibited the proliferation and migration of PC-3 cells (Ps<0.05).

CONCLUSIONThe lentiviral vectors of ILK knockdown and mda7 over-expression have been successfully constructed and identified. The recombinant lentivirus can efficiently infect human prostate cancer PC-3 cells, in which ILK expression is inhibited and mda7 is over-expressed.

Cell Line ; Genetic Vectors ; Humans ; Interleukins ; genetics ; Lentivirus ; genetics ; Plasmids ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo establish a polyethylene glycol (PEG6000) precipitation method for screening macroprolactinemia in patients with high serum prolactin (PRL).

METHODSPEG6000 precipitation method was used to remove macroprolactin (MPRL) molecules in serum of PRL-elevated patients. The effect of PEG6000 precipitating serum MPRL was determined by Sephacryl S-100HR chromatography plus chemiluminescent immunoassay and SDS-PAGE plus Western Blot assay. The PEG6000 precipitation plus chemiluminescent immunoassay was applied to screen serum samples of PRL-elevated patients for macroprolactinemia. The clinical manifestations of patients with true-hyperprolactinemia, hyperprolactinemia/macroprolactinemia or true-macroprolactinemia were analyzed and compared.

RESULTSAfter precipitation with PEG6000, MPRL peak or hybridization signal in the serum samples was markedly decreased, while the big or small prolactin (BPRL or SPRL) levels were not affected. In 1538 PRL-elevated patients, 16.1% (247/1538) were detectable for macroprolactinemia, while the 83.9% (1291/1538) were identified as true-hyperprolactinemia. In 247 samples of macroprolactinemia, 93.5% (231/247) were determined as true-macroprolactinemia, while 6.5% (16/247) were identified as hyperprolactinemia plus macroprolactinemia. In 508 true-hyperprolactinemia patients, menoxenia, menolipsis/menostasia, dysgenesia or hypophysoma were manifested in 438 (86.2%), which were also manifested in 85.7% (6/7) of hyperprolactinemia/macroprolactinemia patients. However, only 11 cases in 71 true-macroprolactinemia patients (15.5%) presented above clinical diseases.

CONCLUSIONThere is a certain proportion of true-macroprolactinemia (pseudo-hyperprolactinemia) in serum PRL-elevated patients. The PEG6000 precipitation method established in this study can efficiently distinguish true-hyperprolactinemia from pseudo-hyperprolactinemia in patients.

Adolescent ; Adult ; Aged ; Female ; Humans ; Hyperprolactinemia ; blood ; diagnosis ; Middle Aged ; Polyethylene Glycols ; Prolactin ; blood ; Young Adult

OBJECTIVETo assess changes of liver function in HIV-positive children with/without HBV/ HCV co-infection after 1 year of highly active antiretroviral therapy (HARRT).

METHODSSeventy-eight pediatric AIDS patients with HBV/HCV co-infection,19 pediatric AIDS patients with HBV co-infection and 44 pediatric AIDS patients without HBV/HCV co-infection who received HAART at least for 1 year were enrolled. HIV-1 viral load was quantitatively detected using a standardized reverse transcriptase-polymerase chain reaction assay, and blood cells were determined by three-color flow cytometry. Anti-HCV antibody and HBsAg was detected using an enzyme-linked immunosorbent technique, and ALT, AST and TBIL were detected by automatic biochemical analyzer.

RESULTSAfter 1 year-HAART, the viral load was decreased to the lowest limit of detection in 90.34% patients (t=2.61, P<0.01), and CD4+ T cell counts were increased from 170.187±132.405/ μl to 796.014±158.491/ μl (t=3.17, P<0.01). The levels of ALT and AST were elevated (t=2.02, P<0.05), while the ALT and AST levels in patients receiving nevirapine (NVP) based HAART increased from 18.28±13.74 U/L and 24.23±8.09 U/L to 55.35±22.40 U/L and 69.97±26.72 U/L, respectively(t=3.80,t=4.11;Ps<0.01). The increment of ALT and AST in NVP based HAART were significantly higher than that in the efavirenz based HAART (ALT:46.28±13.35 U/L vs 37.70±15.25 U/L and AST:19.53±7.23 U/L vs 1.25±0.21 U/L, respectively; t=4.53, t=5.79; Ps<0.01), particularly in patients co-infected with HIV/HBV/HCV (ALT:54.32±22.85 U/L vs 16.89±14.42 U/L and AST:41.71±19.26 U/L vs -3.44±15.59 U/L, respectively; t=3.42, t=2.98, Ps<0.01).

CONCLUSIONHARRT can repress HIV-1 replication effectively, but it also cause the damage of liver function, especially in patients with HBV and/or HCV co-infection.

Antiretroviral Therapy, Highly Active ; Child ; Coinfection ; drug therapy ; Female ; HIV Infections ; complications ; drug therapy ; physiopathology ; Hepatitis B ; complications ; Hepatitis C ; complications ; Humans ; Liver ; physiopathology ; Male

OBJECTIVETo purify polyphenols from Sabina vulgaris and to investigate its antioxidant properties.

METHODSPolyphenols were purified from Sabina vulgaris Antoine with macroporous resin HPD-700, and the quantity of polyphenols was determined by Folin-Ciocalteu colorimetry. The antioxidant properties of polyphenols were evaluated by total antioxidant capacity (T-AOC) and its activities of scavenging DPPH (1,1 diphenyl-2-picry-hydrazyl) radicals, superoxide anion (O2·-), hydroxyl free radicals (OH·) and ferric reducing antioxidant power (FRAP).

RESULTSAfter purification, the purity of polyphenols increased from 0.053% to 0.995%.The antioxidant properties study showed that its inhibition rate of scavenging DPPH radicals and FRAP was 151.83 U/ml and 204.59 U/ml. Its scavenging capacity for superoxide anion (O2·-) and hydroxyl free radicals (OH·) was 151.83 U/ml and 204.59 U/ml. The total antioxidant capacity was 72.68 U/ml.

CONCLUSIONPolyphenols from Sabina vulgaris Antoine have high antioxidant properties, suggesting that it worth further study of its pharmacological effects.

Antioxidants ; pharmacology ; Cupressaceae ; chemistry ; Polyphenols ; isolation & purification

OBJECTIVETo construct the vectors of human glutathione S-transferase A1 (GSTA1), P1 (GSTP1), T1(GSTT1) genes and express in Escherichia coli (E. coli).

METHODSHuman GSTA1, GSTP1 and GSTT1 gene whole length cDNAs were amplified by RT-PCR and then subcloned into pET-28a(+) vectors. The proteins were expressed in E. coli BL21(DE3). After purified by Ni2+ affinity chromatography, the enzymatic activities of GSTs were measured with 1-chloro-2,4 -dinitrobenzene (CDNB) as substrate.

RESULTSThe correct GSTA1, GSTP1 and GSTT1 genes were cloned. And soluble GSTA1, GSTT1, GSTP1 proteins were expressed in E.coli. After purification, GSTA1, GSTT1 and GSTP1 showed good enzymatic activities, which were 17.55, 0.02, 18.75 μmol·min-1·mg-1, respectively.

CONCLUSIONThe expression plasmids for GSTA1, GSTT1 and GSTP1 have been constructed and the recombinant proteins are expressed successfully.

DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; Glutathione S-Transferase pi ; biosynthesis ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Recombinant Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo determine the contents of L-enantiomer impurity in valaciclovir hydrochloride.

METHODSValaciclovir enantiomers were separated and determined by using chiral high performance liquid chromatography. Chromatographic conditions were as follows:CROWNPAK(®) CR(+) chiral column (4 mm×150 mm, 5 μm), detection wavelength:254 nm, mobile phase:water-methanol-perchloric acid (19:1:0.1), flow rate:0.75 ml/min, sample injection volume:10 μl.

RESULTSD-valaciclovir was completely separated from L-enantiomer impurity. The contents of L-enantiomer impurity were 0.65%-2.62% on average in 8 batches of valaciclovir hydrochloride.

CONCLUSIONEnantiomeric impurity contents in each batch of products were all meet criteria of United States Pharmacopeia, which can be used in criteria of Chinese Pharmacopeia as references.

Acyclovir ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Stereoisomerism ; Valine ; analogs & derivatives ; analysis