A high active soybean isoflavone glucoside hydrolase-producing mould strain was isolated from spirit qu. Its optimal hydrolase-producing conditions were as follows: 2.5% wheat bran as carbon source, 1% NaNO3 as nitrogen source, initial pH7. 0, culture medium volume 40mL/250mL, inoculating quantity 8% , culture temperature 30℃, revolutions 160r/min and culture time 84h. The enzyme activity reached 82 U/mL. Cu2+ can inhibit Absidia sp. R strain from producing the hydrolase, the influence of other metal ions was not remarkable on it.

Humans ; Laparoscopy ; Proctectomy ; Rectal Neoplasms/surgery* ; Rectum/surgery* ; Transanal Endoscopic Surgery

Background The retina microglia play a eliminating effect on apoptotie cells in the neural retinal layer of normal rats during postnatal development.Milk fat globule epidermal growth factor 8(MFG.E8)can combine specifically with phosphatidylinositol serine of the surface of apoptotie cells and enhance macrophage phagoeytosis of apoptotic cells.Objective Present study was to evaluate the localization and expression of MFG-E8 and its relevant cytokines in the neural retinal layer of normal rats during postnatal development Methods Normal royal college of surgeon(RCS)rats were divided into P0,P3,P7,Pi4,P30,P45 groups according to their postnatal days,and the 30-day-old RCS rats(2 rats)served as controls.Double stain of M FG.E8 and microglial cells marker(CD11b)was performed by immunofluorescence.Expressions of MFG-E8,integrin β5,CD11b and interleukin-6(IL-6)mRNA in the neural retina were analyzed by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).The utilization of animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State and Science and Technology Commission.Results MFG-E8 and CD11b were positively co-expressed in retinal ganglion cell layer and external plexiform layer with the green fluorescence for FITC-labeled IgG and red fluorescence for cy3-labeled lgG respectively in normal adult rats.RT-PCR showed that the mRNA of MFG-E8,integrin 85,CD11b and IL-6 was detectable at P0 rats.The expression level of these eytokines began to rise fterward and reached peak value at P14 rats and then declined gradually,showing significant differences among different ages groups in various cytokines mRNA expression(all P<0.05).Conclusion MFG-E8 can be specifically expressed in the neural layer of retina microglia in RCS rat.

Animals ; Cells, Cultured ; Female ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Lead ; pharmacokinetics ; toxicity ; Liver ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

OBJECTIVETo explore the clinical application of mandibular-driven simultaneous maxillo-mandihular distraction to correct hemifacial microsomia with rapid prototyping technology.

METHODSThe patient' s skull resin model was manufactured with rapid prototyping technology. The osteotomy was designed on skull resin model. According to the preoperative design, the patients underwent Le Fort I osteotomy and mandibular ramus osteotomy. The internal mandible distractor was embedded onto the osteotomy position. The occlusal titanium pin was implanted. Distraction were carried out by mandibular-driven simultaneous maxillo-mandihular distraction 5 days after operation.

RESULTSThe distraction in five patients was complete as designed. No infection and dysosteogenesis happened. The longest distance of distraction was 28 mm, and the shortest distance was 16 mm. The facial asymmetry deformity was significantly improved at the end of distraction. The ocelusal plane of patients obviously improved.

CONCLUSIONSRapid prototyping technology is helpful to design precisely osteotomy before operation. Mandibular-driven simultaneous maxillo-mandibular distraction can correct hemifacial microsomia. It is worth to clinical application.

Face ; abnormalities ; surgery ; Facial Asymmetry ; congenital ; surgery ; Goldenhar Syndrome ; surgery ; Humans ; Hyperplasia ; surgery ; Mandible ; surgery ; Maxilla ; surgery ; Osteogenesis, Distraction ; methods ; Osteotomy ; methods ; Osteotomy, Le Fort

OBJECTIVETo study the genotoxicity of components of diesel engine exhausts with ethanol-diesel blending fuel. To provide scientific arguments to find more economical and less polluted fuels.

METHODSAmes test, comet assay and GC-MS technique were used to test the genotoxicity and 16 kinds of PAHs on diesel engine exhausts with different proportions of ethanol (E0, E5, E10, E20).

RESULTSBoth Ames test and comet assay were positive. It shows that diesel engine exhausts can lead to mutation and DNA damage, especially in pure diesel oil. But the content of 16 kinds of PAHs and DNA damage level decreased in exhausts of E5. With the increase of ethanol proportion in diesel oil, the content of 16 kinds of PAHs and DNA damage level increased.

CONCLUSIONCompared with pure diesel oil and high proportion of ethanol fuel, E5 can reduce the genotoxicity and the brake specific exhausts of PAHs.

Air Pollutants ; toxicity ; Air Pollution ; Carbon Monoxide ; Comet Assay ; DNA Damage ; Ethanol ; toxicity ; Gasoline ; toxicity ; Mutagenicity Tests ; Particulate Matter ; Vehicle Emissions ; toxicity

Objectives To explore the effects of aging on midgastric transverse band(MTB)and gastric emptying.Methods In our prospective study,57 healthy volunteer were divided into young, middle and old-age groups.After taking test meal labeled by 99mTc-iethylenetriaminepentaacetic acid(99mTc-DTPA), the pictures were collected using double probe single photon emission computed tomography(SPECT).Stomach in each frame of the pictures was divided into proximal, midgastric transverse band(MTB)and distal parts.And half gastric emptying time,gastric remnant rate at 90 min, areas of different gastric parts were tested and calculated respectively.Results Half gastric emptying time of whole stomach was(43.24± 11.87)min,(42.07 ± + 9.31)min and(45.81 ± 10.73)min in young,middle and old-age groups, respectively, with P＞0.05.Among young, middle and oldage groups, half gastric emptying time was(38.09 ± 10.26)min,(37.33 ± 9.28)min and(26.74 ± 12.07)min in proximal stomach, and it was(38.35 ± 12.96)min,(37.73 ± 7.46)min and(46.41 ± 10.74)min in distal stomach,respectively.The half gastric emptying time was significantly reduced in proximal stomach and increased in distal stomach in old-age group(both P＜0.05).The gastric nuclide remnant rate at 90min in total stomach was(30.38 ± 19.32)％,(29.03 ± 10.36)％ and(31.92 ± 13.47)％ ,in young, middle and old-age groups, respectively, with P＞0.05.This rate in proximal stomach was(25.01 ± 12.35)％,(26.36± 15.29)％ and(19.54±8.47)％ among three group, respectively.The rate in proximal stomach was(42.25 ± 12.36)％,(41.56 ±± 9.33)％ and(56.05 ± 11.72)％ among three group,respectively.The gastric remnant rate at 90min was reduced in proximal stomach and was increased in distal stomach significantly in old-age group(both P＜0.05).Compared with young and middle-age group,the old-age group showed no difference in areas of total stomach in all the time, while the areas were reduced in proximal stomach and increased in distal stomach significantly from 30 min to 90 min(all P＜0.05).Total stomach versus proximal and distal stomach showed no difference in count/pixel ratio in all time,while a count/pixel ratio was reduced in proximal stomach and increased in distal stomach significantly from 30 min to 90 min(both P＜ 0.05).Both areas and count/pixel ratio of MTB at 60 min and 90 min were significantly increased in old-age group(both P＜0.05).Conclusions The total gastric emptying is not delayed along with aging, while the gastric emptying is increased in proximal stomach and reduced in distal stomach in the elderly.This abnormity of intragastric distribution of food might be related with larger area of midgastric transverse band.

The 1.7 kb full-length hemagglutinin (HA) gene fragment of H5N1 subtype avian influenza virus was amplified by RT-PCR and then cloned into the pFastBacHT donor plasmids. The recombinant plasmid pFastBac-H5 was identified by restriction enzyme digestion and sequencing. Following the transposition pFastBac-H5 into the bacmid in DH10Bac E.coli competent cells, the colonies were identified by blue and white selection. The recombinant bacmid (rBacmid-H5) was verified by PCR analysis. Transfection of rBacmid-H5 DNA into sf9 cells using Cellfectin reagent results in the production of recombinant viral stock. Cells were harvested 72h post infection and analyzed by SDS-PAGE, Western-blot, hemagglutination and hemagglutination inhibition test;the expressed HA protein (rH5)shows hemaggluting activity and can be inhibited by H5N1 virus immunized chicken sera. On the other hand, immunization of chickens with rH5 protein results in high titers of H5N1 virus specific hemagglutation inhibition antibodies, which proved its biological activity.

This study examined the methylation difference in AIRE and RASSF1A between maternal and placental DNA, and the implication of this difference in the identification of free fetal DNA in maternal plasma and in prenatal diagnosis of trisomy 21. Maternal plasma samples were collected from 388 singleton pregnancies, and placental or chorionic villus tissues from 112 of them. Methylation-specific PCR (MSP) and methylation-sensitive restriction enzyme digestion followed by fluorescent quantitative PCR (MSRE + PCR) were employed to detect the maternal-fetal methylation difference in AIRE and RASSF1A. Diagnosis of trisomy 21 was established according to the ratio of fetal-specific AIRE to RASSF1A in maternal plasma. Both methods confirmed that AIRE and RASSF1A were hypomethylated in maternal blood cells but hypermethylated in placental or chorionic villus tissues. Moreover, the differential methylation for each locus could be seen during the whole pregnant period. The positive rates of fetal AIRE and RASSF1A in maternal plasma were found to be 78.1% and 82.1% by MSP and 94.8% and 96.9% by MSRE + PCR. MSRE + PCR was superior to MSP in the identification of fetal-specific hypermethylated sequences (P<0.05). Based on the data from 266 euploidy pregnancies, the 95% reference interval of the fetal AIRE/RASSF1A ratio in maternal plasma was 0.33-1.77, which was taken as the reference value for determining the numbers of fetal chromosome 21 in 102 pregnancies. The accuracy rate in 98 euploidy pregnancies was 96.9% (95/98). Three of the four trisomy 21 pregnancies were confirmed with this method. It was concluded that hypermethylated AIRE and RASSF1A may serve as fetal-specific markers for the identification of fetal DNA in maternal plasma and may be used for noninvasive prenatal diagnosis of trisomy 21.

This study examined the impact of 935MHz phone-simulating electromagnetic radiation on embryo implantation of pregnant mice. Each 7-week-old Kunming (KM) female white mouse was set up with a KM male mouse in a single cage for mating overnight after induction of ovulation. In the first three days of pregnancy, the pregnant mice was exposed to electromagnetic radiation at low-intensity (150 μW/cm(2), ranging from 130 to 200 μW/cm(2), for 2- or 4-h exposure every day), mid-intensity (570 μW/cm(2), ranging from 400 to 700 μW/cm(2), for 2- or 4-h exposure every day) or high-intensity (1400 μW/cm(2), ranging from 1200 to 1500 μW/cm(2), for 2- or 4-h exposure every day), respectively. On the day 4 after gestation (known as the window of murine embryo implantation), the endometrium was collected and the suspension of endometrial glandular cells was made. Laser scanning microscopy was employed to detect the mitochondrial membrane potential and intracellular calcium ion concentration. In high-intensity, 2- and 4-h groups, mitochondrial membrane potential of endometrial glandular cells was significantly lower than that in the normal control group (P<0.05). The calcium ion concentration was increased in low-intensity 2-h group but decreased in high-intensity 4-h group as compared with the normal control group (P<0.05). However, no significant difference was found in mitochondrial membrane potential of endometrial glandular cells between low- or mid-intensity groups and the normal control group, indicating stronger intensity of the electromagnetic radiation and longer length of the radiation are required to inflict a remarkable functional and structural damage to mitochondrial membrane. Our data demonstrated that electromagnetic radiation with a 935-MHz phone for 4 h conspicuously decreased mitochondrial membrane potential and lowered the calcium ion concentration of endometrial glandular cells. It is suggested that high-intensity electromagnetic radiation is very likely to induce the death of embryonic cells and decrease the chance of their implantation, thereby posing a high risk to pregnancy.