The main commercial production of fructooligosaccharides (FOS) comes from enzymatic transformation using sucrose as substrate by microbial enzyme fructosyltransferase. A fructosyltransferase genomic DNA was isolated from Aspergillus niger QU10 by PCR. The nucleotide sequence showed a 1 941 bp size, and has been submitted to GenBank (KF699529). The cDNA of the fructosyltransferase, containing an open reading frame of 1 887 bp, was further cloned by RT-PCR. The fructosyltransferase gene from Aspergillus niger was functionally expressed both in Escherichia coli and Pichia pastoris GS 115. The highest activity value for the construction with the α-factor signal peptide reached 431 U/mL after 3 days of incubation. The recombinant enzyme is extensively glycosylated, and the active form is probably represented by a homodimer with an apparent molecular mass of 200 kDa as judged from mobility in seminative PAGE gels. The extracellular recombinant enzyme converted sucrose mostly to FOS, mainly 1-kestose and nystose, liberating glucose. FOS reached a maximal value and represented about 58% of total sugars present in the reaction mixture after 4 h reaction. The results suggest that the availability of recombinant Pichia pastoris as a new source of a FOS-producing enzyme might result of biotechnology interest for industrial application.
Aspergillus niger ; enzymology ; genetics ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Fungal Proteins ; genetics ; metabolism ; Glycosylation ; Hexosyltransferases ; genetics ; metabolism ; Molecular Sequence Data ; Molecular Weight ; Pichia ; Sucrose ; metabolism ; Trisaccharides ; metabolism

Objective To explore the clinical value ofmulti-slice spiral CT (MSCT) to carcinomatous intestinal obstruction of colon.Methods The clinical and MSCT data of 40 patients with carcinomatous intestinal obstruction of colon diagnosed by MSCT were analyzed.Results In the 40 cases,complete intestinal obstruction were in 17 cases,incomplete intestinal obstruction in 23 cases,39 cases were carcinomatous intestinal obstruction of colon confirmed by surgery and pathology,and 1 case was intussusception intestinal obstruction by multiple polyps.The MSCT qualitative analysis of obstruction reason was 97.5％ (39/40),the MSCT diagnostic accuracy of intestinal obstruction was 100.0％ (40/40).The obstruction site was at ascending colon showed in 14 cases,transverse colon in 4 cases,splenic flexure of colon in 2 cases,descending colon in 6 cases,sigmoid colon in 12 cases,both ascending colon and transverse colon in 1 case,colon and small intestine in 1 case.The obstruction site accuracy rate of MSCT was 100.0％ (40/40).Conclusion MSCT has obvious advantages in obstruction existing,obstruction site,obstruction reason to carcinomatous intestinal obstruction of colon,it can offer reliable examination data to clinic.

Objective To explore the curative effect of ursodeoxycholi acid on retinopathy of prematurity (ROP) caused by high concentration of oxygen in newborn rats. Methods The model of ROP was established. Neonatal rats were divided into normal control group, ROP model group, low dose ursodeoxycholi acid treatment group (10mg/kg) and high dose ursodeoxycholi acid treatment group (40mg/kg). Rats were sacrificed at days 17. The new retinal vessels were observed and counted under lfuorescence microscope. Results The new retinal vessels in ROP rats were hyperplastic, twisted and unevenly distributed. There was signiifcant difference in the number of new retinal vessels among different groups (P=0.000). The number of new retinal vessels of rats in ROP group, low-dose group and high-dose group was signiifcantly more than that in control group (P=0.000). The number of new retinal vessels in low-dose group and high-dose group was significantly less than that in ROP group (P<0.05). The number of new retinal vessels in high-dose group was less than that in low-dose group (P>0.05). Conclusions Ursodeoxycholi acid could inhibit the angiogenesis of retina and could have curative effect on ROP.

Objective To observe the effect of acute hypervolemic hemodilution(AHH) combined with nicardipine controlled hypotension on hemodynamics changes in revision operation of total hip replacement.Methods Forty patients were divided into two groups according to the sequence of the surgery(odd number came into group A,even number came into group B,20 in each group).In group A,AHH was carried out with nicardipine controlled hypotension;group B was control group.AHH was carried out in both groups before anesthesia induction.6%voluven was transfused in group A at 15 mL?kg-1 and 30 mL?min-1 and nicardipine was transfused at 0.3-0.5 mL?kg-1?h-1 with MAP altering between 60 and 65 mmHg;Same quantity of physiological saline was transfused in group B.HR,MAP,CVP,Hb,Hct and Plt were recorded and observed at preoperation,1 h after AHH and 24 h after the operation.Results In group A after AHH with nicardipine controlled hypotension,MAP was obviously lower than pre-operation(P

Objective To investigate clinicopathological features of scalp angiosarcoma, and to analyze the relationship of tumor stage and treatment with prognosis. Methods Clinical and follow-up data were collected from 16 patients with non-metastatic primary scalp angiosarcoma treated in the Department of Plastic and Reconstructive Surgery of Zhongda Hospital, Southeast University from September 2002 to June 2013. According to the seventh edition American Joint Committee on Cancer (AJCC)TNM staging system for soft tissue sarcomas (2010), staging of scalp angiosarcoma was performed for the 16 patients. Statistical analysis was carried out by the Kaplan-Meier method for survival rates and by the Log-rank test for survival curve. The Cox regression model was used for multivariate regression analysis. Results Of the 16 patients, 4 had stageⅠangiosarcoma, 4 stage Ⅱangiosarcoma, and 8 stage Ⅲ angiosarcoma. The tumor usually began as ecchymosis-like lesions on the head or face in early stage, and progressed into dark red infiltrative plaques, nodules and ulcers which easily ruptured and bled in later stage. Histopathological examination showed generalized vascular proliferation and infiltration with high histological morphological diversity. Cytologic atypia was commonly seen. The median time to recurrence was 15 months, and local recurrence occurred in 7 patients. The median time to metastasis was 20.5 months, and distant metastasis was observed in 8 cases, including 4 cases of pulmonary metastasis, 2 lymph node metastasis, 1 liver metastasis and 1 bone metastasis. The survival time was 33.0 ± 4.4 months (median, 32 months)in 4 patients with stage Ⅰangiosarcoma, 24.0 ± 7.9 months(median, 15 months)in 4 patients with stage Ⅱangiosarcoma, and 23.9 ± 3.9 months (median, 24 months)in 8 patients with stage Ⅲ angiosarcoma. Additionally, the survival time was 23.4 ± 5.2 months(median, 21 months), 24.4 ± 5.7 months(median, 24 months)and 35.8 ± 9.7 months(median, 26 months) in 5 patients receiving surgical treatment alone, 7 patients receiving surgical treatment and radiotherapy, and 4 patients receiving surgical treatment, radiotherapy and immunotherapy, respectively. Conclusions Surgical treatment combined with radiotherapy and immunotherapy may serve as the first-choice treatment for scalp angiosarcoma. Tumor size and treatment regimens are main factors influencing prognosis of scalp angiosarcoma.

BACKGROUND:Total knee arthroplasty had been generaly accepted as the final treatment plan, relieving pain and reconstructing function of knee joint. However, whether drainage tube can be used after replacement is stil controversial.
OBJECTIVE:To compare the clinical effects of drainageversusnondrainage after primary unilateral total knee arthroplasty.
METHODS:Total 102 patients undergoing primary unilateral total knee arthroplasty were randomly divided into 2 groups. In the drainagegroiup, a drainage tube was used. In the nondrainage group,
drainage tube was not used. Total blood loss was calculated by recording the hemoglobin and hematocrit before operation and that after 1, 3, 7 days of operation. The pain visual analogue scale scores, arthrocele, ecchymosis, infection rate, and deep venous thrombosis of lower extremity were examined and analyzed postoperatively. Knee Society Scores were recorded at 1 year postoperatively. Above indexes were compared between the two groups.
RESULTS AND CONCLUSION:(1) Total blood loss and blood transfusion rate were significantly higher in the drainage group than in the nondrainage group (P< 0.05). (2) There was no statisticaly significant difference in the pain visual analogue scale scores, arthrocele, ecchymosis, infection rate, and deep venous thrombosis of lower extremity between the two groups after replacement (P> 0.05). (3) No significant difference in Knee Society Scores was detected between the two groups (P> 0.05). (4) Results indicated that the total blood loss and blood transfusion rate may decrease significantly in patients without wound drainage after total knee arthroplasty. Limb sweling and ecchymosis area were not increased. No significant difference in infection, deep venousthrombosis of lower extremity and knee function was detectable between the two groups. Thus, total knee arthroplasty without wound drainage is safe and does not have obvious adverse consequences.

Objective To investigate the expressions and relationship between lysine-specific demethylase 1 (LSD1) and E-cadherin protein in gastric cancer tissues and adjacent normal tissues,and the correlation with the clinicopathological features and prognosis of patients with gastric cancer.Methods The case-control study was adopted.The gastric cancer tissues and adjacent normal tissues were collected by surgical resection from 80 patients with gastric cancer who were admitted to the Xiangya Hospital of Central South University from June 2008 to June 2009.Expressions of LSD1 and E-cadherin protein were detected by immunohistochemistry (IHC).The follow-up of telephone interview was performed to detect survival of patients till June 2015.Relationships between the expressions of LSD1 and E-cadherin protein and clinicopathological features or prognosis of patients were analyzed.Comparison of count data and correlation were analyzed by the chi-square test and Spearman rank correlation analysis.Survival curve was drawn using the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Results Expressions of LSD1 in cancer tissues and adjacent normal tissues were located at the cell nucleus.The positive expression rate of LSD1 was 67.5％ (54/80) and 43.8％ (35/80) in cancer tissues and adjacent normal tissues,respectively,with a significant difference (x2=9.141,P ＜ 0.05).Expressions of E-cadherin protein in cancer tissues and adjacent normal tissues were located at the cell membrane and cytoplasm.The positive expression rate of E-cadherin was 63.8％ (51/80) and 81.3％ (65/80) in cancer tissues and adjacent normal tissues,respectively,with a significant difference (x2 =6.140,P ＜ 0.05).The positive expression rate of LSD1 was 83.3％ (25/30),76.0％ (19/25) and 40.0％ (10/25) in the low-,moderate-and high-differentiated tumors,37.5％ (6/16),72.7％ (16/22),71.9％ (23/32) and 90.0％ (9/10) in the Ⅰ,Ⅱ,Ⅲ and Ⅳ stages of TNM stage,36.4％ (4/11) and 72.5 ％ (50/69) in patients without and with lymph node metastasis,respectively,showing significant differences (x2=12.870,9.425,4.111,P ＜ 0.05).The positive expression rate of E-cadherin protein was 53.3％ (16/30),56.0％ (14/25) and 84.0％ (21/25) in the low-,moderate-and high-differentiated tumors,75.0％ (12/16),63.6％ (14/22),71.9％ (23/32) and 20.0％ (2/10) in the Ⅰ,Ⅱ,Ⅲ and Ⅳ stages of TNM stage,100.0％ (11/11) and 58.0％ (40/69) in patients without and with lymph node metastasis,70.0％ (49/70) and 20.0％ (2/10) in patients without and with distant metastasis,respectively,showing significant differences (/x2 =6.494,10.073,5.547,7.426,P ＜ 0.05).There was a negative correlation in expressions between LSD1 and E-cadherin protein (r =-0.355,P ＜ 0.05).The survival time and 5-year overall survival rate of patients with positive and negative expressions of LSD1 were (36.9 ± 2.5) months and 31.1％,(47.4 ± 3.4) months and 56.0％,respectively,showing a significant difference (x2=4.550,P ＜0.05).The survival time and 5-year overall survival rate of the patients with positive and negative expressions of E-cadherin protein were (44.0 ± 2.5) months and 46.7％,(32.6 ± 3.5) months and 24.9％,respectively,showing a significant difference (x2 =7.306,P ＜ 0.05).Conclusions Positive expression of LSD1 in gastric cancer tissues is higher than that in adjacent normal tissues.There are increased expression of LSD1 and reduced expression of E-cadherin protein in low-differentiated gastric cancer tissues and high TNM stage,showing a negative correlation between them.Positive expression of LSD1 and negative expression of E-cadherin protein may indicate a poor prognosis.

Objective To explore the impacts of enriched environment(EE),which has different initiation time points and intensity,on the neural and ethological prognosis and contents of myelin basic protein(MBP)of neo-natal rats with hypoxic - ischemic brain damage(HIBD). Methods HIBD rat models were established. Rats were divided into the early,the intermediate and the late intervention groups,which experienced EE from 7,14 and 21 days after HIBD for 14 days. The early and intermediate intervention groups were then divided into 6 - h and 24 - h groups, which experienced EE intervention for 6 hours or 24 hours respectively each day. Trapeze tests and water maze tests were carried out to detect the neural and ethological prognosis. Immunohistochemical method was used to detect MBP of the brain white matter,and the percentages of positive cells with MBP were detected by an image analyzer. The con-tents of MBP were measured. Results The trapeze test scores of the early and intermediate sham operation group,HI group,early 6 - h and 24 - h EE groups and the intermediate 6 - h and 24 - h EE groups,the late sham operation group,the late HI and late EE intervention group were(4. 05 ± 0. 88)scores,(2. 35 ± 1. 02)scores,(3. 67 ± 1. 12) scores,(3. 50 ± 1. 41)scores,(3. 50 ± 0. 93)scores,(3. 56 ± 1. 13)scores,(4. 00 ± 0. 89)scores,(2. 17 ± 1. 17)scores,(3. 50 ± 0. 92)scores,respectively. The trapeze test scores of early,intermediate and late EE groups were higher than those of the HI groups in the same period. There was no significant difference between the early,the intermediate 6 - h EE groups and 24 - h EE groups. Scores of water maze of each corresponding group were(40. 68 ± 23. 77)seconds,(56. 66 ± 10. 96)seconds,(46. 49 ± 19. 27)seconds,(51. 72 ± 20. 46)seconds,(38. 20 ± 18. 36)seconds,(47. 96 ± 20. 65)seconds,(38. 63 ± 20. 44)seconds,(59. 66 ± 13. 81)seconds and(45. 93 ± 22. 45)seconds,respectively. The water maze scores of the early,the intermediate 6 - h EE group and the late 24 -h EE groups were higher than those of the HI groups in the same period. There was no significant difference between the early,the intermediate 6 - h EE groups and the 24 - h EE groups. The relative abundance of MBP of the early and intermediate and the late HI groups were 6. 32 ± 1. 63 and 6. 74 ± 2. 19,and significantly less than that of the sham groups in the same periods,which were 9. 09 ± 1. 69 and 9. 37 ± 2. 46. The relative abundance of MBP of early 6 - h and 24 - h groups,the intermediate 6 - h and 24 - h groups and the late EE group was 7. 84 ± 2. 51,8. 05 ± 1. 86, 8. 89 ± 2. 29,8. 48 ± 2. 67 and 7. 98 ± 2. 09,respectively,which was significantly higher than that of the HI groups in the same periods. It showed that the neural and ethological prognosis of neonatal rats with HIBD could be improved,no matter the intervention began in the early,the intermediate or the late periods,or the intervention time was 6 hours or 24 hours each day. And relative abundance of MBP in the white matter increased with EE. Conclusions EE interven-tion has a long window stage for young rats. EE intervention could improve the neural and ethological prognosis of rats with HIBD. EE intervention could elevate the contents of MBP in the white matter,which could be one of the mecha-nisms for EE to improve the neural and ethological prognosis of rats with HIBD.

Objective To explore the characteristics of illness attribution of outpatients with multiple somatic symptoms in Peking Union Medical College Hospital. Methods It was a cross-sectional study conducted from March to October,2012. A total of 150 outpatients were recruited from the departments of Gastroenterology,Traditional Chinese Medicine and Psychological Medicine by convenience sampling. Somatic symptom scale of the Patient Health Questionnaire (PHQ-15) was used to screening each patient in the waiting list. With the cut-off value of 10,patients were divided into the somatic symptom positive (SOM+) group and somatic symptom negative (SOM-) group. Sociodemographic characteristics were compared between these two groups. All the subjects completed interviews including questions about illness attribution. All the answers of illness attribution were concluded into three major groups as physical factors,situational factors and psychological factors. Results The proportion of female was significantly higher in SOM+ group than in SOM-group (69.3% vs. 53.3%;χ=4.048,P=0.044). In SOM+ group,significantly more patients contributed their illness to psychological factors (64.0% vs. 45.0%;χ=5.273,P=0.022). There was no significantly difference between SOM+ group and SOM-group on the phenomenon of multiple illness attribution (41.0% vs. 32.0%;χ=1.407,P=0.236). However,in the Department of Gastroenterology,there were significantly more outpatients in SOM+ group with multiple illness attribution (60.0% vs. 32.0%;χ=3.945,P=0.047).Conclusions The outpatients in general hospital with multiple somatic symptoms are more likely to contribute their illness to psychological factors. The phenomenon of multiple illness attribution is common among patients. Clinicians should increase their awareness and knowledge of illness attribution,so as to provide better holistic health services.

Objective To explore the influence of early rapidly increased amino acid dosaging on the nitrogen balance and growth in preterm infants.Methods A total of 37 prematures admitted to our NICU within 24 hours after birth and with birth weight of 1 000 g to 1 800 g from November 2011 to January 2013 were randomly divided into study group (n =18) and control group (n =19).Parenteral nutrition (PE) was provided in both groups within 24 hours after birth.The starting amino acid dose was 1.5g/kg per day,then rose by 1 g/kg per day in study group and by 0.5 g/kg per day in control group,until to maximum of 3.5 g/kg per day.The dosage of lipids,glucose,and electrolytes in parenteral nutrition was provided routinely.The trial was double-blinded.Results In the first 5 days of PE,the nitrogen balance had no significant difference between these two groups (F =0.025,P =0.652) ; however,the time of regaining birth weight was significantly shorter in study group [(7.23±2.48) d] than in control group [(9.42±2.03) d] (t=-2.090,P=0.044).There were no differences between these two groups in trems of completely feeding time,weight gain rate,head circumference growth rate,length of hospital stay,and hospital costs (all P ＞ 0.05).Meanwhile,the rates of neonatal necrotizing enterocolitis,sepsis,mechanical ventilation administration,and damage of hepatic and renal function were also not significantly different.The administration of different dosages of amino acids did not affect the energy intake in both groups (F =1.893,P =0.218).Conclusions Prematures need early amino acid administration,with a starting dosage of more than 1.5 g/(kg · d).Rapid increase of amino acid dosing will not affect early nitrogen balance in premature infants; meanwhile,it contributes to weight gain without increasing the complications and hospitalization costs.