Two parental strains BY-14 and BY-6,with high leavening ability in lean and sweet dough respectively,were selected.Through spore production and separation,two haploids with opposition types were selected for cross-breeding.At last one hybridization strain was obtained,with good fermentation ability as BY-14 in lean dough and better than BY-6 by 25%in sweet dough.

Cryptococcosis ; complications ; Eosinophilia ; etiology ; Humans ; Lung Diseases, Fungal ; complications

Child ; Critical Care ; Humans ; Intensive Care Units, Pediatric ; Medical Records ; Severity of Illness Index

Background Vitronectin is a glycoprotein that has a variety of functions.Its expression was markedly higher in the retina of oxygen induced mice,which was confirmed in our animal model,and also increased in human umbilical vein endothelial cells (人 UVECs) that were cultured in high glucose.However,there was no evidence that showed vitronectin was involved in retinal neovascularization.Objective This study was to observe the influence of vitronectin on cytoskeleton remodeling,cell migration and blood vessel formation in 人 UVECs conditioned by high glucose.Methods 人 UVECs were cultured in high glucose and the expression of vitronectin was knocked down using RNA interference technology.The experiments were divided into the high glucose group (人 UVECs were conditioned with DMEM medium that contained 50 mmol/L glucose),negative interference group (人 UVECs were transfected with control siRNA in advance,and then were conditioned with DMEM medium that contained 50 mmol/L glucose) and positive interference group (HUVEC were transfected with vitronectin siRNA in advance,and then were conditioned with DMEM medium that contained 50 mmol/L glucose).The protein expression of vitronectin was measured by Western blot,and the microfilament cytoskeleton of 人 UVECs was examined by immunofluorescence cytochemical staining followed by fluorescence microscopy.Cell migration ability in a scratch wound assay and blood vessel formation ability in a matrigel assay of 人 UVECs were evaluated.The general differences were analysed by One-Way ANOVA ;further contrasts of the two groups were analysed by the LSD-t test.Results The differences in vitronectin expression of the three groups were not obvious at 0 hour (F=1.064,P＞0.05).After 24 hours,vitronectin expression was highest in the high glucose group,lower in the negative interference group,and the lowest in the positive interference group,and the differences were significant (F =15.519,P＜0.05).After 48 hours,vitronectin expression of the three groups displayed the same pattern,and the differences were also significant (F=37.521,P＜0.05).Immunofluorescence showed that the cytoskeleton structure was most obvious in the high glucose group,moderate in the negative interference group,and was the least obvious in the positive interference group,after both 24 hours and 48 hours.In the scratch wound assay,the cell migration ability of the high glucose group was the highest,lower in the negative interference group,and the lowest in the positive interference group after 24 hours,and the differences were significant (F=90.685,P＜0.05).After 48 hours,the cell migration abilities of the three groups displayed the same pattern,and the differences were also significant (F=67.880,P＜0.05).In the matrigel assay,after 6 hours,the number of blood vessels formed in the high glucose group was more than that in the negative interference group,and the least amount was found in the positive interference group.The differences among of them were significant (F =86.653,P＜0.05).The number of blood vessel formed in the positive interference group was also the lowest after 12 hours,and the differences were also significant (F=18.992,P＜0.05).Conclusions Vitronectin can bring about cytoskeleton remodeling,increase in cell migration,and enhancement of blood vessel formation in 人 UVECs conditioned in high glucose.It may be one of the important influence factors of diabetic retinopathy.

Objective To evaluate the efficacy and safety of diacerhein in the treatment of patients with knee osteoarthritis.Methods The efficacy and safety of diacerhein was randomly investigated in 42 pa- tients with knee osteoarthritis using parallel group methodology and a double-dummy technique to ensure dou- ble blind status with respect to diacerhein and control drugs diclofenac.Results Significant changes were ob- served in 20 meters walk pain,knee joint tenderness scale,WOMAC index scale,5F-36 health survey,knee joint swelling scale,compared with baseline(P＞0.05)in both diacerhein and diclofenac group respectively.No difference was found between diacerhein group and diclofenae group.The patient global assessment and physi- cian's global assessment were similar in diacerhein group and diclofenac group(P＞0.05).The side effect was similar in two groups.All of these side effects in gastrointestinal tract appeared to be transient.Conclusion Diacerhein can effectively relieve pain and swelling of knee osteoarthritis,and provides us a new effective and safe approach for treating knee osteoarthritis.

Objective To investigate the significance and effect of ultrasonic diagnosis on the autologous bone marrow mesenchymal stem cells (MSC) in angiogenesis. Methods Twenty-four New Zealand rabbits were divided into experiment group (12) and the control group (12). Then rabbit bone marrow MSCs from experiment group were isolated, caltured and marked with Brdu. After ischemic hind limb animal model on all rabbits was set up, autologous bone marrow MSCs were directly injected into the ischemic hind limb muscles in experiment group while same volume normal saline was used in the control group. Two weeks after the implantation of autologous bone marrow MSCs, 2D and color Doppler flow imaging (CDFI) detection were used in rabbit femoral artery of the two groups to observe the inner diameter of the blood vessel, the peak velocity and the acceleration time. The disposition of transplaned cells and the state of angiogenesis in ischemic muscles were assessed using immunofluorescence staining. Results The results of 2D and Doppler ultrasound detection showed the inner diameter of the blood vessel and the peak velocity of the blood current in experiment group obviously higher than that of the control group , and the acceleration time was obviously smaller than that of the control group P<0.01. The immunofluorescence staining showed there were transplanted cells existed in transplanted portion and state of angiogenesis was supurior obviously than that of the control. Conclusions Bone marrow MSCs had the effect to promote angiogenesis. Implantation of autologous bone marrow MSCs was a simple and efficient therapeutic method for the ischemia hind limb. Using high-frequency ultrasound to detect femoral artery may provide a practical and useful method to evaluate the effect on implanted autologous bone marrow mesenchymal stem cells.

ObjectiveTo observe the effect of progesterone on progesterone receptor in brain tissue with hypoxia-ischemia brain da-mage(HIBD) in neonatal rats,and discuss the protective mechanism of progesterone on HIBD of neonatal rats.MethodsTwenty-four 7-day-old neonatal rats were randomly divided into 3 groups:sham-operated group,hypoxia-ischemia group and pretreatment group.Rats in hypoxia-ischemia group and pretreatment group were subjected to left common carotid artery ligation,then were exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in 37 ℃ closed container for up to 2.5 h to establish hypoxia-ischemia encephalopathy(HIE) model.Progestero-ne was injected intraperitoneally into pretreatment group rats respectively at 30 minutes before hypoxia,solution was injected into the first 2 groups.All rats were killed at the 24 hour after operation,immunohistochemistry staining was used to examine the expression of progesterone receptor in brain.ResultsTotally 24 neonatal rats entered the result analysis without loss.Progesterone receptor was expressed in both endochylema and nucleus in every group rat.The amount of the positive cell of progesterone receptor in hypoxia-ischemia group significantly decreased compared with that in sham-operated group(P

CTB protein possessed mucosal adjuvant immunoactivity. The CTB gene was amplified by PCR method from a strain V. cholerae. The nucleotide sequence of CTB gene was 375 bp and shared 96.0%~99.2% homology with other 6 CTB genes. The recombinant plasmid pTWIN1-CTB transformed E. coli strain BL21(DE3) expressed with 0.8 mmol/L IPTG. The molecular weight of expression products was identical with expectative weight by SDS-PAGE electrophoresis. The CTB fusion proteins mainly assembled inclusion bodies and the outputs of proteins were approximately 20% of the total bacterial proteins. The CTB proteins possessed mucosal immunoactivity by GM1-ELISA assay.

Background Researches determined that the alteration of A69S locus of age-related maculopathy susceptibility 2 ( ARMS2 ) gene is closely associated with the pathogenesis and progression of age-related maculopathy ( AM D ).However,the location of ARMS2 protein in normal eye tissue is still in controversy,therefore,its function is below understanding up to now.Objective The goal of this laboratory work was to investigate the distribution,expression and location of ARMS2 protein in normal adult retina and choroid as well as in retinal pigment epithelial (RPE) cells and lay a basis for exploring further its function in the protein level.Methods Ten donor eyeballs of normal adult male with the age from 28-42 years were collected in eye bank of Qingdao Eye Hospital.The frozen sections of the retina and choroid were prepared for the detection and location of ARMS2 in 3 eyes by immunofluorescence under the confocal laser microscope.The retina was isolated for the primary culture of RPE cells using explant culture method.The cells were then identified by CK32 antibody by immunofluorescence.The distribution and expression of the ARMS2 protein in retina,ehoroid and RPE cells were determined by immunofluorescence technique.Results ARMS2 protein was strongly expressed in retinal vessel,RPE cell layer,Bruch membrane and choroidal vessel,but weak expression was in retinal ganglion cell layer,inner nuclear layer,outer plexiform layer,outer nuclear layer and inner plexiform layer in the normal eyes.The primarily cultured cells appeared the polygon shape with the abundant pigment in cytoplasm.The immunofluorescence of the cells showed the positive response for CK32,exhibiting the green fluorescence granules in the cytoplasm.The positive expression of ARMS2 protein also was seen in the cytoplasm of RPE cells,appearing the red fluorescence.Conclusions ARMS2 protein mainly distribute and locate retinal and choroidal vessels,RPE cells and Bruch membrane in normal eye.

Objective To investigate the protective effect of insulin-like growth factor-1 (IGF-1) on 6-hydroxy dopamine (6-OHDA) induced oxidative damage in PC-12 cells. Methods PC12 cells were treated with 6-OHDA (concentrations of 25, 50, 100, 150 and 200 μmol/L). In order to select the optimal experimental concentration and treatment time, the activity of PC12 cells was detected by MTT at different time points of 12 h, 24 h and 48 h after treatment. PC12 cells were divided into three groups: control group, 6-OHDA group and IGF-1+6-OHDA group. The activity of PC12 cells was detected by MTT assay. Reactive oxygen species (ROS) level of PC12 cells was detected by immunofluorescence staining, and apoptosis was detected by Hoechst33342 / PI double staining method. Results With the increased concentration and the prolongation of the action time of 6-OHDA, the activity of PC12 cells decreased gradually. The concentration of 150 μmol/L and action time of 24 h of 6-OHDA were selected as the optimal experimental concentration and observation time for this study. Compared with 6-OHDA group, the activity of PC12 cells increased, the expression level of ROS and the apoptosis decreased in IGF-1+6-OHDA group. Conclusion IGF-1 pretreatment can reduce 6-OHDA induced oxidative damage and apoptosis of PC12 cells, also can increase cell activity, which can provide a potential strategy for the prevention and treatment of Parkinson’s disease.