To study the alkaloid constituents in the seed of Sophora alopecuroides L. and isolation of lehmannine by transhydrogenation of sophocarbine to matrine present in the mixture. Methods　Various processes of exchange, ion exchange and column chromatograph with very similar physical and chemical properties was successfully separated by transhydrogenation. Results　5 alkaloids were obtained and identified by elementary analysis, IR, MS, 1HNMR and 13CNMR as oxymatrine, oxysophoridine, matrine, sophoridine and lehmannine (12, 13-dehydromatrine). Conclusion　Lehmannine was isolated from the seed of S. alopecuroides for the first time.

Object To study the alkaloid constituents in the seed of Sophora alopecuroides L and isolation of lehmannine by transhydrogenation of sophocarbine to matrine present in the mixture Methods Various processes of exchange, ion exchange and column chromatograph with very similar physical and chemical properties was successfully separated by transhydrogenation Results 5 alkaloids were obtained and identified by elementary analysis, IR, MS, 1HNMR and 13 CNMR as oxymatrine, oxysophoridine, matrine, sophoridine and lehmannine (12, 13 dehydromatrine) Conclusion Lehmannine was isolated from the seed of S alopecuroides for the first time

Objective To study the pathogenesis of anemia in chronic aristolochic acid nephmpathy(CAAN) rats. Methods The hemoglobin(Hb)values of sixty-two male SD rats were assayed to determine its normal range.Among them,24 rats with normal Hb value were randomly divided into 2 groups:model group (MG)in which rats received the extract of Aristololochia manshuriensis Kom (AmK) by gavage,and control group (CG) received tap water only by gavage.Body weisht(BW),Hb,24 h urinary protein excretion(UP)and creatinine clearance (Ccr)of 6 rats in each group were measured before administration and at the end of the 8th week, respeetively.then these rats were sacrificed.The relative area of renal interstitial fibrosis was measured by microscopy.The mRNA expression of erythropoietin (EPO)in kidney tissue Was determined by real-time RT-PCR;protein expression of type I collagen(Coll),aminopeptidase P (APP),hypoxia indHeible factor let and 2α(HIF-1α and HIF-2α)in kidney tissue Was examined by immunohistochemistry staining. Results Hb values of normal rats presented normal distribution. The normal Hb was (155.9±16.5) g/L. Rat anemia was diagnosed when Hb was below 123.6 g/L. There was no difference in all the examination results between CG and MG before administration (P>0.05). Compared with CG, the Hb and Cer in MG were significantly decreased [(121.66±15.68) g/L vs (169.00±12.89) g/L, (0.63±0.13) ml/min vs (1.27±0.18) ml/min, P< 0.01], and the UP in MG was significantly increased at the end of the 8th week [(27.04±9.40) mg/d vs (6.11±0.84) mg/d, P<0.01]; the relative areas of fibrosis and Col l in renal interstitium of MG were significantly enlarged [(12.89±2.33)% vs (0.55±0.10)%, (13.92±2.92)% vs (1.32±0.84)%, P<0.01]; the protein expression of APP and the mRNA expression of EPO in the kidney tissue of MG were significantly down-regulated [(0.55±0.23)% vs (3.77±1.06)%, 0.005±0.001 vs 0.032±0.013, P<0.01]; the protein expression of HIF-lα and HIF-2α in the kidney tissue of MG was significantly up-regulated (2.55±0.16 vs 1.12±0.46, 2.33±0.33 vs 1.15±0.27, P<0.01), at the end of the 8th week. Conclusions The pathogenesis of anemia in CAAN may be due to the decreased production of EPO caused by the destruction of peritubular capillary. The compensatory up-regulation of HIF-lα and HIF-2α expression can not prevent the anemia development.

Objective To explore the dynamic changes of diffusion tensor imaging(DTI) in spinal cord of goats with persistent compression injury. Methods Eighteen goats weighted 20--25 kg were divided into three groups with completely random design: A, B and C. A balloon catheter was inserted into the epidural space at C3-4 level via intervertabral foramen for each goat. The balloon was inflated by injection of variable volumes of saline in group A and B 10 days following operation. The volume of saline was 0. 3 ml in group A and 0. 2 ml in group B,respectively. The compression sustained for 40 days. Group C served as uncompressed control without injection of saline. The locomotor rating score was applied to each group. Conventional MRI and DTI were performed. The apparent diffusion coefficient (ADC)and fractional anisotropy (FA) values were measured. Histopathological assessments of the compressed spinal cord were performed 50 days following operation with light microscope and transmission electron microscopy. Results Before operation, the locomotor rating score was 5, the ADC value was ( 1.23 ± 0. 05 ) × 10-3 mm2/s and the FA value was (0. 72 ± 0. 05 ) each group. Of six goats in Group A, the locomotor rating score severely decreased and reached( 1.5±0. 4)on the 40 th day after compression. The ADC value at compression site decreased soon and reached the minimum (0. 75±0. 04) × 10-3mm2/s on the 5 th day after compression. Then the ADC value increased gradually, restored normal on the 10 th day or so, then became markedly higher than normal and reached( 1.61±0. 05) × 10-3mm2/s on the 40 th day. The FA value at compression site decreased soon, reached(0. 54±0. 04)on the 1st day, then decreased gradually and reached(0. 43 ± 0.05) on the 40 th day. It appeared high signal intensity on T2WI on the 10 th day. In Group B, the locomotor rating score was moderately decreased and reached(3.4±0. 5 )on the 40 th day. The ADC value at compression site decreased slightly firstly, reached( 1.08±0. 04) × 10-3mm2/s on the 1st day, restored normal on the 20 th day or so, then increased gradually, became higher than normal and reached ( 1.27 ± 0. 05) × 10-3mm2/s on the 40 th day. The FA value increased slightly firstly, reached (0. 78±0. 05 )on the 1st day, then decreased gradually, restored normal on the 15 th day or so, became lower than normal and reached(0. 67±0. 05) on the 40 th day. There was no abnormality on conventional MRI. In Group C, the locomotor rating score, ADC value and FA value remained unchanged. There was no abnormality on conventional MRI. There were dynamic changes over time of the ADC value and FA value in Group A and B, which was more marked in Group A than that in Group B ( repeated measurements analysis of variance, F=426. 7 for the ADC value and F =7895.2 for the FA value, P < 0. 01 ). Histopathologically, swelling and degeneration of axons and neurons as well as the disarrangement of myelin sheathes could be seen. The pathological changes were more marked in Group A than in Group B. In Group C, no abnormality could be seen. Conclusion There are dynamic changes of DTI in cervical spinal cord with compressive injury that correlated with the degree and duration of compression. The ADC value decreased firstly, restored normal and then increased. The FA value increased firstly, restored normal and then decreased in mild compression while solely decreased in serious compression.

The key challenge to generate engineered cells by synthetic biology for producing 7-dehydrocholesterol (7-DHC) in a high titer is the match between functional module and chassis. Our study focused on solving this problem by combining different promoters and yeast chassis to increase 7-DHC production. To optimize the chassis in order to accumulate zymosterol, the substrate for 7-DHC synthesis, we overexpressed truncated HMG-CoA reductase (tHmglp) and squalene epoxidase (Erglp), both are key genes of yeast endogenous zymosterol biosynthetic pathway. In addition, we knocked out C-24 methyl transferase (Erg6p) and C-22 dehydrogenase (Erg5p) to inhibit the conversion of zymosterol to ergosterol. By introducing heterologous C-24 reductase under three promoters with different strengths, namely TDH3p, PGK1p and TDH1p, we constructed functional modules of diverse activities. Nine engineeredcells were generated based on the combination of these three modules and three chassis. The result shows that the engineered cell composed of functional module regulated by TDH3p and chassis SyBE_000956 had the highest 7-DHC production, indicating a better match than others. This study provides evidences for importance of match and empirical support for rational design of subsequent researches.
Cholesterol ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; Dehydrocholesterols ; metabolism ; Gene Knockout Techniques ; Hydroxymethylglutaryl CoA Reductases ; metabolism ; Industrial Microbiology ; Methyltransferases ; genetics ; Promoter Regions, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; Synthetic Biology

Objective To explore the effects of BSA on hypoxia inducible factor/hypoxia response element (HIF/HRE) transcription activity in rat tubular epithelial cells (NRK-52E) with HRE-Luc reporter plasmid.Methods Luciferin activity of NRK-52E cells incubated by a medium contained BSA in varying concentration (0,5,10,20 mg/ml) and stimulus duration (24,48,72 h) was detected by dual luciferase detecting system based on HRE-Luc reporter plasmid and HIF-1 α expression was detected by Western blotting.Results HIF/HRE transcription activity of NRK-52E cells was increased in BSA incubation group (10 mg/ml,48 h) compared with blank control (BSA 0 mg/ml,48 h) [(2.59±0.35)vs (1.03±0.09),P=0.000].HIF-1α expression of NRK-52E cells was increased in BSA incubation group (20 mg/ml,48 h) compared with blank control (BSA 0 mg/ml,24 h) [(0.052±0.010) vs (0.014±0.003),P=0.000].Conclusion Albumin can increase HIF/HRE transcription activity of TEC.

Objective To assess the value of a fibrin-targeted contrast agent (EP-2104R) for MR detection of thrombus, and to compare this modality with non-contrast-enhanced (NCE) MRI and Gd-DTPA injection at acute period after thrombus generation. Methods Thrombus was induced with external injury and stasis in 5 rabbits. MRI was performed before and after contrast agent injection at 6.0 h after injury, and the MRI findings were compared with that of histopathologically examinations. Results EP-2104R enhanced MRI accurately detected thrombus, which was superior to both NCE and Gd-DTPA injection (P<0.001). Gd-DTPA injection was not associated with improvement of thrombus detection. Conclusion Being a fibrin-targeted MR contrast agent for in vivo detection of acute thrombus, EP-2104R is superior to NCE MRI and Gd-DTPA injection.

Objective To investigate the feasibility of automatic spectral imaging protocol selection (ASIS) and adaptive statiatical iterative reconstruction (ASiR) technique to reduce radiation dose and dose of contrast agent.Methods Sixtyfour patients underwent routine abdominal examination were randomly divided into two groups.The test group used ASIS technique,with 30％ ASiR and 50％ ASiR reconstruction algorithm.The control group used 120 kVp tube voltage,FBP reconstruction method.The noise of liver,pancreas,sacrospinal muscle,CNR of liver and pancreas,subjective image score in arterial phase and portal venous phase were compared between the image of 70 keV+30％ ASIR and control group.CNR of abdominal aorta and its branchs,CNR of portal vein,and subjective image score were statistically analyzed between im age 55 keV+50％ ASiR and control group in the arterial phase and portal venous phase.Results Compared with control group,CT dose index volume for arterial phase and portal venous phase in test group decreased by 23.68％,23.57％ and dose length product decreased by 25.61％,18.45 ％,total contrast injection decreased 16.86 ％,the noise of liver,pancreas and sacrospinal muscle in 70 keV+30％ ASiR were lower than those of control group in abdominal arterial and portal phase (all P＜0.05).CNR of abdominal aorta,superior mesenteric artery,celiac axis and score in 55 keV+50％ ASiR were higher than those of control group in abdominal arterial phase (all P＜0.05),CNR of portal vein and score in portal phase had no statistically difference (all P＞ 0.05).Conclusion Combining of ASIS and ASiR including 70 keV + 30％ ASiR and 55 keV+50％ ASiR,images are superior to that of the conventional 120 kVp+FBP scan mode for abdominal CT image and vessel image quality,which can reduce the radiation dose and the dose of contrast agent.

Objective: To observe the role of Qudu medicinal granules (祛毒冲剂) on enterogenous endotoxemia . Methods: Sixty-one cases with enterogenous endotoxemia were randomly divided into two groups:Qudu medicinal granule group (n=30) that was treated with Qudu medicinal granules combined with western medicine, and smecta group which was treated with smecta and western medicine (n=31). Changes of symptoms and signs were observed before treatment and 1, 3, 7 days after treatment. Blood samples were collected in the morning to measure the white blood cell (WBC), plasma lipopolysaccharide (LPS) and tumor necrosis factor (TNF) levels. Results: Recovery speed of WBC count in Qudu medicinal granule group was faster than that of the smecta group, there was significant difference on the third day after treatment (P