The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
Amino Acid Sequence ; Chromatography, High Pressure Liquid ; methods ; Interferons ; standards ; Mass Spectrometry ; methods ; Molecular Weight ; Oxidation-Reduction ; Peptide Mapping ; Protein Processing, Post-Translational ; Reference Standards

OBJECTIVETo identify and determine the congener and level of microcystins in the source water of Taihu Lake.

METHODSImproved method of SPE combined with HPLC was employed to detect the concentration and varieties of microcystins in source water and bloom samples collected from Meiliang Bay, Taihu Lake.

RESULTSThe contents of two predominant microcystin components, MC-RR, and MC-LR, were relatively high in samples during warm months and correlated with the phase of algae growth. The maximum concentrations of MC-RR and MC-LR in water sample reached 3.09 +/- 0.53 microg/L and 2.39 +/- 0.41 microg/L during the period of water bloom in September 2004, respectively. Even without waterbloom, the concentration of MC-LR in source water sample was still higher than the guideline value.

CONCLUSIONThe status of microcystin pollution in this region is serious and measures to monitor and control the growth of cyanobacteria are urgently needed.

Animals ; Bacterial Toxins ; analysis ; China ; Chromatography, High Pressure Liquid ; Cyanobacteria ; growth & development ; isolation & purification ; Environmental Monitoring ; Fresh Water ; analysis ; chemistry ; Microcystins ; analysis ; chemistry ; Spectrophotometry, Ultraviolet ; Time Factors ; Water Microbiology ; Water Pollutants, Chemical

Objective To investigate the relationship between the expression of matrix metalloproteinase-9 (MMP-9) and intercellular adhesion molecule-1 (ICAM-1) in the perihematomal tissues in human hypertensive,intracerebral hemorrhage (ICH) and brain edema formation following ICH.Methods Paraffin-embedded brain tissues of 39 human fatal cases of ICH from the perihematomal tissues,1—3 cm away from the margin of the hemorrhagic lesion,as well as tissues from the corresponding area at the opposite side as controls,were stained with HE and immunohistochemistry staining.The expressions of MMP-9 and ICAM-1 in the pefihematomal tissues were analyzed with the SPSS 11.5 system.Results ①With MMP-9 immunohistochemical staining positive capillaries in the perihematomal tissues were identified at 2 h ((1.2? 0.8)/HP).The number of MMP-9 positive capillaries began to rise at 5—10 h ((4.1?0.8)/HP) reaching the peak at 45—48 h ((10.6?1.4)/HP,P

Child ; Child, Preschool ; Cryptorchidism ; diagnosis ; surgery ; Humans ; Infant ; Laparoscopy ; methods ; Male

AIM To explore the clinical effects of Yiqi Huoluluo Mingmu Decoction combined with Conbercept Ophthalmic Injection on patients with non-proliferative diabetic retinopathy.METHODS One hundred and sixty-seven patients were randomly assigned into control group(83 cases)for administration of Conbercept Ophthalmic Injection,and observation group(84 cases)for administration of both Yiqi Huoluluo Mingmu Decoction and Conbercept Ophthalmic Injection.The changes in clinical effects,miR-15b,CTGF,VEGF,hemodynamic indices(PSV,EDV,RI),visual field gray value,CMT,hemangioma count,blood spot area,BCVA,electroretinograms(amplitudes of a wave,b wave under light-dark adaptation),TCM symptom scores and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed increased miR-15b,PSV,EDV,BCVA,electroretinograms(P＜0.05),and increased CTGF,VEGF,RI,visual field gray value,CMT,hemangioma count,blood spot area,TCM symptom scores(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with non-proliferative diabetic retinopathy,Yiqi Huoluluo Mingmu Decoction combined with Conbercept Ophthalmic Injection can improve hemodynamics,reduce fundus lesions,and regulate miR-15b,VEGF expressions.

OBJECTIVEWe identify ionizing radiation-induced mitochondrial DNA (mtDNA) deletions in human lymphocytes and their distribution in normal populations.

METHODSLong-range polymerase chain reactions (PCR) using two pairs of primers specific for the human mitochondrial genome were used to analyze the lymphoblastoid cell line following exposure to 10 Gy (60)Co γ-rays. Limited-condition PCR, cloning and sequencing techniques were applied to verify the mtDNA deletions detected with long-range PCR. Human peripheral blood samples were irradiated with 0, 2 and 6 Gy (60)Co γ-rays, and real-time PCR analysis was performed to validate the mtDNA deletions. In order to know the distribution of mtDNA deletions in normal population, 222 healthy Chinese adults were also investigated.

RESULTSTwo mtDNA deletions, a 7455-bp deletion (nt475-nt7929 in heavy strand) and a 9225-bp deletion (nt7714 -nt369 in heavy strand), occurring between two 8-bp direct repeats, were identified in lymphoblastoid cells using long-range PCR, limited-condition PCR and sequencing. These results were also observed for (60)Co γ-rays irradiated human peripheral blood cells.

CONCLUSIONTwo novel mtDNA deletions, a 7455-bp deletion and a 9225-bp deletion, were induced by ionizing radiation. The rate of the mtDNA deletions within a normal population was related to the donors' age, but was independent of gender.

Cell Line ; Cloning, Molecular ; Cobalt Radioisotopes ; DNA Damage ; genetics ; radiation effects ; DNA, Mitochondrial ; genetics ; radiation effects ; Gene Deletion ; Genetic Markers ; Humans ; Lymphocytes ; radiation effects ; Radiation, Ionizing ; Real-Time Polymerase Chain Reaction

The column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds from the EtOAc extract of medium and MeOH extract of cell cultures of Morus alba. Eight compounds were isolated. Based on physico-chemical properties and spectroscopic data, their structures were identified as isobavachalcone (1), genistein (2), norartocarpetin (3), albanin A (4), guangsangon E (5), mulberrofuran F (6), chalcomoracin (7), kuwanon J (8). Compounds 3-6 were isolated from the cell cultures of M. alba for the first time.
Benzofurans ; isolation & purification ; Cell Culture Techniques ; methods ; Chalcones ; isolation & purification ; Chromatography, Gel ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Genistein ; isolation & purification ; Morus ; chemistry ; cytology ; Plant Leaves ; chemistry ; cytology ; Plants, Medicinal ; chemistry ; cytology ; Silica Gel ; Terpenes ; isolation & purification

The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.
Amino Acid Sequence ; Chromatography, High Pressure Liquid ; Molecular Sequence Data ; Peptide Mapping ; Recombinant Fusion Proteins ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tandem Mass Spectrometry ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; chemistry

Recent randomized controlled clinical trials in Africa have demonstrated that adult male circumcision (MC) efficiently decreases the rate of HIV, HPV and HSV-2 infections. Many studies have clearly shown that MC is a simple, safe, and cost-effective method for the prevention of sexually transmitted diseases and urinary tract infection, and for improving genital hygiene. While a 30% MC prevalence exists worldwide, only 5% or less of the Chinese males have undergone circumcision. In this review, we report recent trends in international MC and HIV prevention efforts, as well as the potential benefits and importance of promoting MC in China. We appeal to medical and public health authorities to pay close attention to the international experience in MC and HIV prevention.
Circumcision, Male ; HIV Infections ; prevention & control ; Humans ; Male ; Reproductive Medicine ; Sexually Transmitted Diseases ; prevention & control

OBJECTIVETo establish a novel and useful rabbit model of lumbar disc degeneration using microinjection of fibronectin fragment (Fn-f).

METHODSThirty-two New Zealand white rabbits underwent injection of N-terminal 30 kDa Fn-f (experimental group) or phosphate buffered saline (PBS) (control group) into the central region of L1-2, L2-3, L3-4, L4-5 discs using a 32-gauge microsyringe. Two rabbits (blank group) with no treatments were sacrificed to examine the proteoglycan synthesis of neucleus pulposus (NP) using (35)S-sulfate incorporation assay. At the 4-, 8-, 12-, and 16-week time points, the discs were examined histologically, radiographically, and with proteoglycan synthesis.

RESULTSHistology demonstrated a progressive loss of the cell numbers in NP and architecture destruction in NP and anulus fibrosus (AF) in Fn-f-injected discs over the 16-week study period. The NP regions in Fn-f-injected discs shrinked distinctly after the 4-week time point, and were not discernible with the inner AF by the 16-week time point. Protoglycan synthesis in Fn-f-injected discs decreased progressively (F = 263.241, P = 0.000). At each time point, the Fn-f-injected discs showed significantly decreased proteoglycan synthesis compared with controls (t = -27.010 - -2.833, P < 0.05). The DHI% of the Fn-f-injected discs at the 4-, 8-, 12-, and 16-week time points were 96.5% ± 1.7%, 85.6% ± 3.8%, 77.2% ± 3.5% and 65.5% ± 5.6%, respectively. Comparing with the DHI% of PBS-injected discs (97.4% ± 1.2%), the Fn-f-injected discs exihibited no significant differences in disc heights at the 4-week time point (P > 0.05), but significant decreases in disc heights at the 8-, 12-, and 16-week time points (t = -21.225 - -10.795, P < 0.01). Apparent anterior osteophytes formed at the 12-week time point and enlarged remarkablely by the 16-week time point in the experimental spines.

CONCLUSIONSFn-f can induce a progressively degenerative process in rabbit discs which is ethical, cost-effective, reproducible, and consistent with the spontaneous degeneration in human. And it seem to be a novel and useful model for the study of disc degeneration at the molecular level.

Animals ; Disease Models, Animal ; Fibronectins ; pharmacology ; Intervertebral Disc Degeneration ; chemically induced ; Lumbar Vertebrae ; Rabbits ; Random Allocation