Two high protease producing strains, whic h are desi gnated Aspergillus oryzae AS100 and AS102, were screened and applied as main fermentation strains of swine blood meal. A Saccharomyces cerevisiae Y113 and a Bacillus Asp007 were also used as assistant fermentation strains. The enzyme production abilities we re studied,and a series of parameters of the fermentation technology were deter mined. Through swine blood meal fermentation, a high-protein fermented feed wa s produced, which is strong soy flavor and was rich in protein(69%), free amino acid, vitamin such as VitD 3 and niacin and organic elements such as Fe. It is a kind of high-protein feed for animals and it could be used as feedstuff addi tive.

Objective To observe the expressions of bone morphogenetic protein-2(BMP-2)and bone morphogenetie protein-7(BMP-7)in the synovial tissue of fluorosis rats and its correlation with pathogenic mechanism of fluorosis arthritis.Methods Thirty-two SD rats were randomly divided into 4 groups:the control group,low,moderate and high-dose fluoride group.The control group ate commou fodder.The low,moderate and high dose fluoride group were fed with fodder composed of 25%.35%and 68%of corn(containing fluorine of 148.00 mg/kg)in chronic endemic fluorosis region in Guizhou Province.After 140 days,the expressions of BMP-2 and BMP-7 protein were determined by immunohistochemistry and assayed the absorbanee by computer image-pattern analysis system.Light microscope was used to observe the synovial tissue by Hematoxin Eosin,and calculated the pathological integral of synovium according to pathological grade standard.Results The expressions of BMP-2 (32.50±2.73)and BMP-7(38.90±2.56)in the control group was spare.Compared with the control group,the expressions of BMP-2(59.43±5.12,79.82±6.41,101.76±7.56)and BMP-7(55.10±4.82,78.42±5.61,98.46± 6.05)in the synovial tissue was up-regulated in each experimental groups(P＜0.05),especially in the moderate dose and the high-dose groups(P＜0.05).Compared with the control group(0.54±0.21).the pathological integral of synovium increased(P＜0.05)in each experimental groups(1.04±0.98,4.69±1.28,8.60±2.07).The expressions of BMP-2 and BMP-7 in the synovial tissue was found to be positively related with the pathological integral of synovium(r=0.98,0.99,P＜0.05).Conclusion The BMP-2 and BMP-7 play an important role in the development of fluorosis arthritis,probably by affecting osteogenesis.

Objective To investigate the relationship between hemoglobin and blood pressure of pregnant women in Zhoushan islands, so as to provide scientific evidence for the etiological study of gestational hypertension. Methods A retrospective study was conducted among 1 383 pregnant women who received perinatal care in Zhoushan Maternal and Child Health Hospital of Zhejiang Province from January 2017 to June 2018. Pregnant women were monitored for hemoglobin content and blood pressure in the early, middle and late pregnancy. The multivariate linear regression was used to analyze the relationship between hemoglobin content and blood pressure in different pregnancy. Results The incidence of anemia in early, middle and late pregnancy was 7.74%, 25.45% and 15.76% respectively. The multivariate linear regression showed that hemoglobin levels during pregnancy had effects on systolic blood pressure in early, middle and late pregnancy, and the earlier hemoglobin levels were monitored, the more obvious the effect on systolic blood pressure was.With the increase of hemoglobin level, systolic blood pressure increased, such as the effect of hemoglobin on systolic blood pressure in early pregnancy, mid-pregnancy and late pregnancy. Hemoglobin of first trimster had the greatest effect (β=0.10, P<0.001), Hemoglobin of second trimester had no obvious effect, and that of third trimester had the second effect (β=0.04, P=0.027).Hemoglobin levels and diastolic blood pressure levels were similar to their relationship with systolic blood pressure. Conclusions Hemoglobin levels during pregnancy have significant effects on systolic and diastolic blood pressure in first, second and third trimsters of pregnancy. Regular measurement of hemoglobin levels during pregnancy can improve the health of pregnant women.

OBJECTIVETo isolate and purify exosomes derived from human bladder transitional cell carcinoma T24 cells, analyze the morphology and protein composition, and investigate the antitumor effect of specific cytotoxic T lymphocytes induced by exosomes.

METHODSExosomes were isolated and purified by ultrafiltration and sucrose gradient centrifugation, and characterized by electron microscopy and Western blot. Dendritic cells were amplified and purified from peripheral blood and pulsed with exosomes. Then they were co-cultured with T cells, and divided into 3 groups: exosome-pulsed DC group, unplused DC group and control group. Alamar-Blue assay was used to evaluate the specific cytolytic activity.

RESULTSThe exosomes were in size about 30 approximately 90 nm saucer-shaped membranous vesicles. HSP70, ICAM-1 and CK20 were detected by Western blot. The CTL induced by DC pulsed with exosomes had significant cytolytic activity (P < 0.01).

CONCLUSIONThe exosomes derived from T24 cells are loaded with immunoprotein HSP70 and ICAM-1, and DC pulsed with exosomes can promote the anti-tumor effect of CTLs in vitro.

Carcinoma, Transitional Cell ; pathology ; Cell Line, Tumor ; Coculture Techniques ; Cytotoxicity, Immunologic ; immunology ; Dendritic Cells ; cytology ; immunology ; ultrastructure ; Exosomes ; immunology ; metabolism ; ultrastructure ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Keratin-20 ; metabolism ; Lymphocyte Activation ; T-Lymphocytes ; cytology ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Urinary Bladder Neoplasms ; pathology

AIM: To evaluate the safety and efficacy of intravitreal bevacizumab ( avastin ) injection in patients with exudative age related macular degeneration ( AMD) .
METHODS: The records of patients treated with intravitreal injection of 1. 75mg bevacizumab for AMD were retrospectively reviewed. All patients were evaluated by complete ophthalmic examination, optical coherence tomography and fundus fluorescein and/or indocyanine green angiography. Observation was made on the best corrected visual acuity ( BCVA) , intraocular pressure, and the changes of lens, vitreous, central retinal thickness (CFT) and total macular volume (TMV), at 1d, 3d, 7d, 1mo and 6mo after the treatment and then compared with those of pre - operation. Repeated treatment with intravitreous bevacizumab occurred if there were signs of persistent or recurrent exudation. And all cases were followed up at least 6mo. An intravitreal injection of bevacizumab (1. 75mg) was given once every 6wk.
RESULTS:All 50 eyes of 48 patients with the average of 58±20. 46 years old were included. The mean baseline of BCVA and CFT were 0. 82±0. 53, and 364. 97±151. 83μm respectively. Although there was no significant decrease in mean CFT and TMV one week after the injection, the mean BCVA had significant improvement. At the last visit of 9. 7mo follow - up, BCVA, CRT and TMV showed significant improvements over baseline values. BCVA was improved by at least two lines in 32 eyes (64%),remained stabilization in 18 eyes (36%) at the last visit. A total of 98 injections were performed and the average number of injections was 1. 98 for each eye in the group. About 50%of re - injections gained at least two lines of vision improvement one week postoperatively. There were no serious adverse events during the treatment.
CONCLUSION: Intravitreal bevacizumab ( avastin ) injection for managing CNV due to age-related macular degeneration is safe and few side effects. Intravitreal avastin associated with improvement in visual acuity ( VA ) , which can reduce macular edema and choroidal neovascularization leakage. But a prolonged treatment effect needs further observation.

To evaluate the safety and efficacy of intravitreal bevacizumab injection in patients with macular edema (ME) induced by retinal vein occlusion (RVO).
● METHODS: The records of patients treated with intravitreal injection of 1. 75mg bevacizumab for ME induced by RVO were retrospectively reviewed. All patients were evaluated by complete ophthalmic examination, optical coherence tomography ( OCT) and fundus fluorescein angiography ( FFA ), etc. Best corrected visual acuity (BCVA), intraocular pressure, the change of lens and vitreous, central foveal thickness (CFT) were observed at 1, 2, 3, 6mo after treatment and compared with before treatment. Repeated treatment with intravitreous bevacizumab occurred if there were signs of persistent or recurrent exudation. All the cases were followed up at least 6mo. An intravitreal injection of bevacizumab (1. 75mg) was given at 6wk intervals.
●RESULTS: Fifty patients (56 eyes) with the average of (57±18. 56) years old were included. The mean baseline of BCVA, CFT were (logMAR0. 82±0. 63), (626. 5±178. 0)μm respectively. Although there was no significant decrease in mean CFT at 1wk after injection, the mean BCVA had significant improvement. Followed up at mean 10. 26 ± 5. 87mo, BCVA, CFT showed significant improvements over baseline values. The statistics of CFT at 1, 2, 3mo after injection were significant differences compared with before injection in each of the three groups. CFT at 1, 3, 12mo after injection were (365. 11±23. 212) μ m, (333. 42± 35. 526) μ m, (267. 6 ± 116. 8) μ m, which had a significant difference ( P < 0. 001), namely macular retinal thickness was thinner obviously that before treatment, ME was improved obviously. CFT was no significant difference at each time point after injection in the group of BRVO-ME and CRVO- ME (P> 0. 05). OCT image showed that after injection macular retinal thickness was becoming thinner. FFA showed that after injection macular fluorescein leakage decreased. BCVA was improved by at least two lines in 48 eyes (86%),remained stable in 8 eyes (14%) at the last visit. A total of 112 injections were performed and the average number of injections was 1. 96 in the group. About 50% of reinjections gained at least two lines of vision improvement at 1wk following the retreatment. There was no serious complications during the treatment.
●CONCLUSlON: lntravitreal injection of bevacizumab can improve visual acuity (VA) of RVO (CRVO and BRVO) in patients with ME, relieve ME, reduce the leakage of CNV, and repeated treatment is better. But a prolonged treatment effect needs further observation. There are no serious ocular and systemic complications occurred in our study.

OBJECTIVEThis study aimed to prepare a hypoxic brain damage model in the neonatal rat using a new approach, 0% oxygen exposure, and to explore the reliability and advantages of the new model.

METHODSSeven-day-old Wistar rats were randomly exposed to either 7 minutes of 0% oxygen, to the conventional Rice-Vannucci method (ischemia + 2 hrs hypoxia exposure), or to left common carotid artery ligation (ischemia). Rat pups which were not subjected to any hypoxia-ischemia treatment were used as the control group. Brain water content and neuronal apoptosis were measured. Neurofunctional assessment was performed. Brain pathological changes were observed using hematoxylin and eosin staining.

RESULTSThe water content (88.96+/-0.29%) and apoptosis of neurons (31.52+/-5.45%) of the left brain in the 0% oxygen group were significantly higher than those of the ischemic and the control groups (P<0.01), and similar to those in the Rice-Vannucci group. The water content (88.68+/-0.24%) and apoptosis of neurons (30.85+/-5.38%) of the right brain in the 0% oxygen group were significantly higher than those of the Rice-Vannucci, the ischemic and the control groups (P<0.01). Both side brains of the 0% oxygen group showed pathological injuries, but only left brain of the Rice-Vannucci group had pathological changes. No pathological abnormalities were seen in the ischemic and the control groups. Significant neurofunctional impairments were found in the 0% oxygen and the Rice-Vannucci groups.

CONCLUSIONSA hypoxic brain damage model of neonatal rat was successfully prepared using 7 minutes 0% oxygen exposure. The new approach appears to be simple and reliable.

Animals ; Animals, Newborn ; Apoptosis ; Brain ; pathology ; Disease Models, Animal ; Female ; Hypoxia, Brain ; pathology ; physiopathology ; Male ; Rats ; Rats, Wistar

BACKGROUNDCisplatin (DDP) is one of most effective and most commonly used therapeutic agent in treating tumors, it can accumulate in the kidney and lead to acute renal failure. MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family. In the present study, we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.

METHODSHK-2 cells were cultured, transfected with microRNA-181a inhibitor for 48 hours, and stimulated with 50 µmol/L cisplatin for 24 hours. MicroRNA-181a expression was analyzed by real time PCR, and cell apoptosis was detected by flow cytometry. Moreover, Bcl-2 and Bcl-2-associated X protein (Bax) expression were measured by Western blotting.

RESULTSMicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor, compared with that in untransfectd cells (21.19 ± 2.01 vs. 38.87 ± 1.97, P < 0.05). Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor. Compared with DDP treated cells alone, Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.

CONCLUSIONOne pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.

Apoptosis ; drug effects ; genetics ; Blotting, Western ; Cell Line ; Cisplatin ; pharmacology ; Flow Cytometry ; Humans ; MicroRNAs ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the effects of particulate matters less than 2.5 μm in aerodynamic diameter (PM2.5) on heart repolarization/depolarization and heart rate variability (HRV).

METHODSWe conducted a panel study for elderly subjects with heart disease in Beijing from 2007 to 2008. PM2.5 was measured at a fixed station for 20 h continuously each day while electrocardiogram (ECG) indexes of 42 subjects were also recorded repeatedly. Meteorological data was obtained from the China Meteorological Data Sharing Service System. A mixed linear regression model was used to estimate the associations between PM2.5 and the ECG indexes. The model was adjusted for age, body mass index, sex, day of the week and meteorology.

RESULTSSignificant adverse effects of PM2.5 on ECG indexes reflecting HRV were observed statistically and the strongest effect of PM2.5 on HRV was on lag 1 day in our study. However, there were no associations between PM2.5 and ECG indexes reflecting heart repolarization/depolarization. Additionally, the effects of PM2.5 on subjects with hypertension were larger than on the subjects without hypertension.

CONCLUSIONThis study showed ambient PM2.5 could affect cardiac autonomic function of the elderly people with heart disease, and subjects with hypertension appeared to be more susceptive to the autonomic dysfunction induced by PM2.5.

Aged ; Air Pollutants ; toxicity ; Electrocardiography ; Environmental Monitoring ; Female ; Heart Diseases ; physiopathology ; Heart Rate ; drug effects ; Heart Ventricles ; drug effects ; physiopathology ; Humans ; Male ; Middle Aged ; Particle Size

BACKGROUNDBlood coagulation factor VII (FVII) is physiologically synthesized in the liver and released into the blood. Binding of FVII to tissue factor (TF) is related to the metastatic potential of tumor cells, also a significant risk factor in the development of hepatic metastasis in patients with colorectal cancer (CRC). It has been found that some cancer cells can produce FVII extrahepatically. However, little is known about FVII and CRC. We therefore hypothesized that CRC cells may synthese FVII, leading to tumor invasion and metastasis.

METHODSWe detected the expression of FVII protein in 55 CRC specimens by immunohistochemical staining. The FVII mRNA in 45 of 55 CRC cases, 6 colon cancer cell lines and one hepatoma cell line was measured by real-time reverse transcription-PCR (RT-PCR). Transwell invasion assays were performed to evaluate the changes of cell migration and invasion of LoVo cancer cells in vitro. We further observed the likely effectors regulated by the TF/FVIIa complex Western blotting assay.

RESULTSExtrahepatic synthesis of FVII was detected in the cytoplasm of 32 (58.2%) CRC specimens by immunohistochemistry, but not in normal mucosa. Liver metastasis (P = 0.003) and TNM staging (P = 0.005) were significantly correlated with FVII antigen expression. The positive ratios in stages I, II, III and IV were 33.3%, 40.0%, 52.4% and 87.5%, respectively. The expression of FVII mRNA in CRC with hepatic metastasis was significantly higher than CRC without hepatic metastasis (5.33 ± 2.88 vs. 1.47 ± 0.51, P = 0.03). Ectopic FVIIa induced a slight increase (1.34-fold) in the number of migrating cells, which was inhibited by the specific TF antibody. The formation of TF/FVIIa complex resulted in a marked increase in the expression of matrix metalloproteinases (MMP)-2 (3.5-fold) and MMP-9 (4.7-fold) in a time-dependent and dose-dependent manner.

CONCLUSIONSExtrahepatic synthesis of FVII by CRC cells may promote tumor invasion and metastasis. MMPs, as downstream effectors of TF/FVIIa signaling, facilitate the development of metastasis in colon cancer.

Adult ; Aged ; Aged, 80 and over ; Cell Line, Tumor ; Cell Movement ; Colorectal Neoplasms ; metabolism ; pathology ; Factor VII ; analysis ; biosynthesis ; genetics ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; secondary ; Male ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; RNA, Messenger ; analysis ; Thromboplastin ; physiology