The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy-transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Förster-type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy.
Animals ; Anti-Bacterial Agents ; chemistry ; metabolism ; pharmacology ; Binding Sites ; Cattle ; Humans ; Kinetics ; Lactoferrin ; chemistry ; metabolism ; Pefloxacin ; chemistry ; metabolism ; pharmacology ; Protein Binding ; Protein Conformation ; Serum Albumin ; chemistry ; metabolism ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet

The effect of L-proline as a promoter on the condensation reaction of salicylaldehyde or its derivatives with ethyl acetoacetate in neutral ionic liquid [emim]BF4 was studied. All reactions were carried out under mild reaction conditions and achieved high yields. Moreover, the ionic liquid containing L-proline could be recycled and reused for several times without noticeably decreasing in productivity. The results show that the L-proline-[emim]BF4 system has a potential in contribution to the development of environmentally friendly and inexpensive processes in organic syntheses.
Coumarins ; chemical synthesis ; Ionic Liquids ; chemistry ; Proline ; chemistry

Objective: To observe the migration of 4,4＇-diaminostilbene-2,2＇-disulfonc acid-based fluorescent whitening agents ( DSD-FWAs ) in food packaging paper, so as to provide evidence for quality and safety supervision for paper packaging materials. Methods: Forty-one paper samples with DSD-FWAs positive were made into 6 cm2 pieces and were soaked in four food simulants ( distilled water, 3% acetic acid, 10% ethanol and 95% ethanol, 10 mL each ). The experiment was carried out at the specified soaking temperature and time. The migration amounts of eleven DSD-FWAs were detected by high performance liquid chromatography-fluorescence detection. Results: C.I.220, C.I.24, C.I.210, C.I.85, C.I.113, C.I.264, C.I.353 and C.I.357 were found in all the four food simulants. At the same time and temperature, the migration amount was highest in 10% ethanol, followed by distilled water, 3% acetic acid and 95% ethanol. C.I.220 was dissolved in all four food simulants, in the range of 20-90 ℃, the migration amount increased with soaking temperature; at 20 ℃, 40 ℃ and 60 ℃, the migration amount increased first and then stabilized over time. Conclusion The higher the storage temperature and the longer the storage time of paper packaging, the easier the DSD-FWAs in packaging paper migrate to food.

OBJECTIVETo evaluate a "silicone suture" technique for enhancing the effect of scalp reduction.

METHODSUnder the local anesthesia, when an incision was made in the midline of the lesion, the dissection was carried out underlying the sub-galea on both sides of the lesion, as far as the width of the lesion. A 3 mm silicone suture in diameter was placed in the galea beyond the lesion. After the first suture bite was anchored in the tissue at one end, the suture device was continued across the midline in such a way as a running, buried, horizontal mattress suture and it was brought out to the skin surface through the deep tissue at the another end of the lesion with a locker. The extra-tissue of the lesion was then excised and the wound was directly closed in layers. After one week of the operation, the silicone suture was gradually tightened in 2-3 times a week for about 3-5 weeks, until both sides of the lesion were approximately closed. The device was there after removed and the wound was directly closed in layers after the scar was excised.

RESULTSBetween October of 1999 and March of 2006, 12 scarring-scalp patients, 7 males and 5 females, were treated by using the silicone suture device without complications. The excised defects were 5-10.5 cm in width. The stretching period was 26.4 days in mean. With the following-ups over 3 months, no hypertrophic scar and widening scar cases appeared.

CONCLUSIONSThe silicone suture as an alternative device for tissue extension could be a safe, simple, effective and economical device. It could significantly enhance the efficiency of scalp reduction.

Adolescent ; Adult ; Alopecia ; etiology ; surgery ; Cicatrix ; complications ; surgery ; Female ; Humans ; Male ; Scalp ; surgery ; Silicones ; Sutures ; Young Adult

OBJECTIVETo observe the level of intracellular reactive oxygen species (ROS) in rats with severe burn and pulmonary microvascular endothelial cells (PMVECs) treated with serum of rat with burn injury, and to investigate the relationship between ROS and apoptosis of PMVECs.

METHODS(1) Twenty-four SD rats were divided into sham injury group ( n = 3) and burn group (n = 21) according to the random number table (the same grouping method below). Rats in burn group were inflicted with 30% TBSA full-thickness scald on the back, and rats in sham injury group were sham injured. Blood samples were collected from abdominal aorta at post injury hour 6, 12, 24, 36, 48, 60, 72 respectively from 3 rats of burn group. The serum content of ROS was assayed by ELISA. The same determination was performed in rats of sham injury group. (2) Five rats were subjected to scald injury as above, and burn serum was prepared 24 hours after injury. Another 5 rats without receiving any treatment were used to prepare normal serum. (3) Marginal pulmonary tissue was harvested from 20 SD young rats. Cells were cultured with tissue block method and indentified with immunohistochemical staining. The third passage of PMVECs in logarithmic phase were inoculated in 6-well plates and 12-well plates. PMVECs in both plates were divided into 4 groups: normal serum group, burn serum group, normal serum + MnTBAP group, and burn serum + MnTBAP group, with 3 wells in each group. Cells in the former 2 groups were respectively cultured with special nutrient solution of endothelial cells without serum added with 15% healthy rat serum or 15% burn rat serum. Cells in the latter 2 groups were cultured with the same culture conditions as in the former two groups correspondingly with addition of 100 µmol/L MnTBAP in the nutrient solution. After being cultured for 24 h, the content of ROS in PMVECs in 6-well plates was detected with flow cytometry. The apoptosis of PMVECs in 12-well plates was observed with acridine orange-ethidium bromide staining, and the apoptosis rate was calculated. Data were processed with one-way analysis of variance and LSD-t test.

RESULTS(1) The serum contents of ROS in rats of burn group were respectively (187 ± 21), (235 ± 22), (231 ± 25), (291 ± 20), (315 ±23) nmol/mL at post injury hour 24, 36, 48, 60, 72, which were significantly higher than that in sham injury group [(141 ± 19) nmol/mL, with t values respectively 7. 86, 9. 57, 13. 87, 14.98, 18.40, P values below 0.01]. (2) Primary cells grew slowly and showed a cobblestone appearance. After passages, cells grew with orderly distribution. The positive rate of coagulation factor VIII of cells was (96 ± 5)% , and thus they were identified as PMVECs. (3) In normal serum group, burn serum group, normal serum + MnTBAP group, and burn serum + MnTBAP group, the contents of ROS in PMVECs were respectively 798 ± 40, 1 294 ± 84, 763 ± 59, 926 ± 42 ( F =93.01, P <0.01), and the apoptosis rates of PMVECs were respectively (6.2 ± 1.3)%, (57.3 ± 6. 7)%, (3.7 ± 0. 8)%, (28.7 ± 5. 7)% (F = 224.50, P <0.01) after being cultured for 24 h. Compared with those of normal serum group, the content of ROS and apoptosis rate of PMVECs in burn serum group increased significantly (with t values respectively 10.40 and 49.06, P values below 0.01). The content of ROS and apoptosis rate of PMVECs in burn serum + MnTBAP group were significantly lower than those in burn serum group (with t values respectively 7.48 and 23.94, P values below 0.01).

CONCLUSIONSSerum content of ROS was increased in severely burned rats. Burn rat serum stimulation on PMVECs can lead to the increase of the intracellular ROS and induce apoptosis. However application of MnTBAP can scavenge ROS and reduce the apoptosis induced by burn rat serum.

Animals ; Apoptosis ; Burns ; blood ; therapy ; Endothelial Cells ; pathology ; Enzyme-Linked Immunosorbent Assay ; Lung ; blood supply ; Oxygen ; Rats ; Reactive Oxygen Species ; blood ; Serum ; metabolism

OBJECTIVETo study the effect of the technique of synchronously perforating and transplanting hair follicular-units in the treatment of cicatricial alopecia after burn.

METHODSOne hundred and sixty-six patients with 217 bald scar areas after burn were treated with above-mentioned technique from January 2002 to April 2008. Scalp strips, with conforming the necessity for grafting, were harvested from the occipital or temporal region. A series of follicular-units, each composing 1 - 3 hairs, were dissected from the strips under microscope or magnifying glass. Size-matching micro-slots were made in the scarred recipient area with 16 - 20 G needles to accept the grafts. The prepared follicular-unit was synchronously implanted into the bottom of the micro-slot as the needle being withdrawn. Patients who were not satisfactory with the density of hairs after I stage surgery underwent II stage surgery a half year later. Ten recipient areas with clear boundary in 10 patients were optionally chosen to observe the density of follicular-units and hair amount with naked eyes after I stage surgery. Survived transplanted hairs in above-mentioned 10 areas were counted to calculate hair survival rate at follow-up. Patients' postoperative satisfaction ratings were surveyed with questionnaire.

RESULTSIn one half of the patients, treatment was finished after I stage surgery, the other one half received 2 stages of surgery. The follicular-unit density reached 15 - 25 grafts/cm(2) with 40 - 70 hairs/cm(2) after I stage surgery. All patients were followed up for over 8 months. Grafted hairs grew well in a natural way. 96.5% mean hair survival rate was observed in the 10 recipient areas. From patients who received only I stage surgery, 61 patients (73.5%) were very satisfactory and 22 patients (26.5%) satisfactory with the results. From the other half of patients, 76 patients (91.6%) were very satisfactory and 7 patients (8.4%) satisfactory with the results.

CONCLUSIONSThe technique of perforating and transplanting follicular-unit hair synchronously is safe and effective with less surgery-induced injury and less bleeding. Hairs transplanted on cicatricial alopecia area with this technique grow well with high survival rate.

Adolescent ; Adult ; Aged ; Alopecia ; etiology ; surgery ; Burns ; complications ; Child ; Cicatrix ; complications ; Female ; Hair Follicle ; transplantation ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Young Adult

The lack of effective in vitro infection model for hepatitis E virus (HEV) has greatly hindered the quantitative analysis of neutralizing titers of anti-HEV antibodies and human sera, thus impeding further studies of HEV-stimulated antibody responses and the immunological mechanisms. In order to improve this situation, the infection of HepG2 cells that are inefficient for HEV replication was continuously monitored until the viral load reached the limit of detection on day 13, the results of which confirmed the feasibility of using this cell line to establish the infection model. Then, neutralization assays of five anti-HEV murine monoclonal antibodies and serum samples collected from four HEV vaccine recipients (collected before and after vaccination) were performed by 96 multi-channel parallel infections, nucleic acid extraction, and qPCR. The results showed that the cell model can be applied for quantitative evaluation of the neutralizing capacity of different antibodies and antiserum samples from HEV vaccine recipients. In this study, we have successfully established a high-throughput in vitro HEV replication model, which will prove to be useful for the evaluation of HEV vaccines and studies of HEV epitopes.
Animals ; Antibodies, Viral ; analysis ; immunology ; Hepatitis Antibodies ; analysis ; immunology ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; chemistry ; immunology ; physiology ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; Mice, Inbred BALB C ; Neutralization Tests ; methods ; Virus Replication

Objective To investigate the effect of wheat-grain size cone moxibustion plus acupuncture on related high-frequency ultrasonographic indicators before and after its treatment of mild to moderate carpal tunnel syndrome and explore its mechanism of action. Method Sixty patients were randomized to an acupuncture group (20 cases) and a wheat-grain size cone moxibustion groups (40 cases). The acupuncture group received acupuncture at affected-side points Daling (PC7), Hegu (LI4), Neiguan (PC6) and Shousanli (LI10). The wheat-grain size cone moxibustion group received wheat-grain size cone moxibustion on point Daling and acupuncture at the other points. Treatment was given once every other day for a total of 10 times. The thickness of the transverse carpal ligament, the cross-sectional area of the median nerve at the level of the pisiform bone and the flattening ratio at the level of the hamate hook were measured by high frequency ultrasonography before and after treatment. The clinical therapeutic effect was evaluated using the SSS score. Result After treatment, the SSS score decreased in both groups compared with before (P<0.05) and was lower in the wheat-grain size cone moxibustion group than in the acupuncture group (P<0.05). The cure and marked efficacy rate was 71.1％ in the wheat-grain size cone moxibustion groupwhich was higher than 42.1％ in the acupuncture group (P<0.05). There were no statistically significant pre-/post-treatment differences in the thickness of the transverse carpal ligament and the flattening ratio at the level of the hamate hook in the two groups (P>0.05). After treatment, the cross-sectional area of the median nerve at the level of the pisiform bone decreased in both groups compared with before (P<0.05) and was smaller in the wheat-grain size cone moxibustion group than in the acupuncture group (P<0.05). Conclusion Wheat-grain size cone moxibustion plus acupuncture can markedly improve the clinical symptoms of carpal tunnel syndrome. Its mechanism of action is related to reducing the cross-sectional area and edema of the median nerve.

The aim of this study is to construct a phage display single-chain variable fragment (scFv) library against breast cancer cells and screen the specific antibodies against MCF-7 cells from the library. The BALB/C mice were immunized with MCF-7 cells. Total RNA of spleens was isolated. The heavy-chain (VH) and light-chain variable region genes (VL) of the antibodies were amplified by RT-PCR and joined into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The assembled scFv fragments were cloned into the phagemids(pCANTAB5E) and the recombinant phagemids were used to transform competent E. coli TG1. The transformed TG1 cells were infected by helper phage M13KO7 and the recombinant phagemids were rescued. The scFv fusion proteins were displayed on the surfaces of the recombinant phages. A phage display antibody library of repertoire of 1.2 x 10(6) clones was constructed. The specific antibodies against MCF-7 cells were enriched by 75 times after five rounds of affinity selection. Ten recombinant phages clones that exhibited specific binding to MCF-7 cells were identified. The specificity of those phage clones was analyzed by reactivity against HepG2 cells and Hela cells by ELISA. One of the selected phage clones against MCF-7cells was used to infect E. coli TOP10 to produce the soluble scFv antibodies after induction with IPTG. The strategy of construction and screening of antibody library directed against the whole tumor cells described in this report should be generally applicable to generate tumor cell-specific antibodies.
Animals ; Antibodies, Neoplasm ; genetics ; immunology ; Breast Neoplasms ; immunology ; therapy ; Cell Line, Tumor ; Female ; HeLa Cells ; Hep G2 Cells ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Single-Chain Antibodies ; genetics ; immunology

OBJECTIVETo investigate the mechanism of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells.

METHODSIn this study, the proteins extracted from paclitaxel-induced apoptotic MCF-7 cells were analyzed by 2-dimentional gel electrophoresis (2-DE), and compared with those from untreated MCF-7 cells. The differential proteins were identified by mass spectrometry.

RESULTSAt 24 hour after paclitaxel (100 nmol/L) treatment, MCF-7 cells were collected and extracted the whole proteins. Seventeen up-regulated or down-regulated proteins were found by analysis of the differential proteomic 2-DE map. Six of them were identified by mass spectrometry. They were enolase 1, chloride intracellular channel 1, keratin 8, ribosomal protein S12, galectin-1 and histidine triad nucleotide binding protein, respectively.

CONCLUSIONWe effectively found the changed proteins in the process of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells by proteomic techniques. These up-regulated or down-regulated proteins are important molecules for our further research about the mechanism of paclitaxel-induced apoptosis.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Female ; Galectin 1 ; metabolism ; Humans ; Keratin-8 ; metabolism ; Paclitaxel ; pharmacology ; Phosphopyruvate Hydratase ; metabolism ; Proteomics ; methods ; Ribosomal Proteins ; metabolism