Analysis on Changes of Aquaporin and Intestinal Flora in a Rat Model with Spleen-Yin Deficiency Syndrome / 中国实验方剂学杂志

ObjectiveTo investigate the effects of spleen-Yin deficiency on gastrointestinal absorption， water metabolism and intestinal flora in rats with spleen-Yin deficiency syndrome. MethodA rat model of spleen-Yin deficiency syndrome was established by using the composite factors， including irregular meat and vegetable diet， weight-bearing fatigue swimming and gavage with warm-heat injury-Yin drugs. The changes of body weight， food intake， water intake and duration of swimming in the blank and model groups were observed. Hematoxylin-eosin（HE） staining was used to observe the histopathological damage of the stomach and colon. Urinary excretion rate of D-xylose was determined by phloroglucinol method. The content of gastrin（GAS） in serum was determined by enzyme-linked immunosorbent assay（ELISA）. The relative expression levels of vasoactive intestinal peptide（VIP）， aquaporin 3（AQP3） and AQP4 in gastric tissues were detected by Western blot. The relative mRNA expression levels of VIP， AQP3 and AQP4 in gastric tissues were detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， and the changes of intestinal flora were analyzed by 16S rDNA sequencing. ResultCompared with the blank group， the results of general physical signs showed that the body weight and food intake of rats in the model group were significantly decreased， the water intake was significantly increased（P<0.05， P<0.01）， and the duration of swimming was significantly decreased（P<0.01）. Pathological examination results showed that in the mucosa of gastric tissues of rats in the model group appeared to be misaligned， the mucosa of colonic tissues could be seen to be obviously thinned or mutilated， and the epithelial cells appeared to be necrotic or even exfoliated. Compared with the blank group， the urinary D-xylose excretion rate of rats in the model group was significantly decreased（P<0.01）， and the serum GAS content was significantly decreased（P<0.05）. Compared with the blank group， Western blot results showed that the relative expression level of VIP protein in gastric tissues of rats in the model group was significantly decreased， while the relative expression levels of AQP4 and AQP3 proteins were significantly increased（P<0.01）. Compared with the blank group， Real-time PCR results showed that the relative expression level of VIP mRNA in gastric tissues of rats in the model group was significantly decreased（P<0.01）， and the relative mRNA expression levels of AQP3 and AQP4 were significantly increased（P<0.05， P<0.01）. Compared with the blank group， the results of intestinal flora analysis showed that the number of operational taxonomic units（OTUs） and α-diversity increased and β-diversity decreased significantly in the model group， the abundance of Porphyromonadaceae was increased significantly， and the abundance of Oscillibacter_ruminantium was decreased significantly（P<0.05）. Spearman correlation analysis showed that Porphyromonadaceae was significantly positively correlated with AQP4 protein level， while Oscillibacter_ruminantium was significantly positively correlated with VIP protein level， and negatively correlated with AQP3 and AQP4 protein levels（P<0.05）. Linear discriminant analysis effect size（LEfSe） analysis results showed that there were significant differences in a variety of intestinal bacteria between groups， and the intestinal bacteria of the model group were significantly enriched in the phylum/order/family/genus of Elusimicrobia， Betaproteobacteria， Burkholderiales， Sutterellaceae and Parasutterella（P<0.05）. ConclusionSpleen-Yin deficiency syndrome can weaken the digestion and absorption capacity of gastrointestinal tract， and cause the disturbance of water metabolism and intestinal flora. AQP4， AQP3 and VIP protein levels of gastric mucosa are closely related to Porphyromonadaceae and Oscillibacter_ruminantium. And AQP4， AQP3 and VIP may be involved in the regulation of intestinal flora in order to affect the physiological function of spleen governing transportation and transformation.

Engineering the glycogen metabolism in cyanobacterial photosynthetic cell factories: a review / 生物工程学报

Cyanobacteria are important photosynthetic autotrophic microorganisms and are considered as one of the most promising microbial chassises for photosynthetic cell factories. Glycogen is the most important natural carbon sink of cyanobacteria, playing important roles in regulating its intracellular carbon distributions. In order to optimize the performances of cyanobacterial photosynthetic cell factories and drive more photosynthetic carbon flow toward the synthesis of desired metabolites, many strategies and approaches have been developed to manipulate the glycogen metabolism in cyanobacteria. However, the disturbances on glycogen metabolism usually cause complex effects on the physiology and metabolism of cyanobacterial cells. Moreover, the effects on synthesis efficiencies of different photosynthetic cell factories usually differ. In this manuscript, we summarized the recent progress on engineering cyanobacterial glycogen metabolism, analyzed and compared the physiological and metabolism effects caused by engineering glycogen metabolism in different cyanobacteria species, and prospected the future trends of this strategy on optimizing cyanobacterial photosynthetic cell factories.

Diagnosis of glutaric acidemia type ⅡC by fetal whole exome sequencing / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis of a fetus with enlargement and enhanced echo of the kidneys.@*METHODS@#The imaging data of the fetus were collected, in addition with 20 mL amniotic fluid sample and 2 mL peripheral blood samples of both parents. Amniotic DNA was extracted for library construction and whole exome sequencing, and Sanger sequencing was carried out to verify candidate variant associated with the fetal phenotype.@*RESULTS@#Prenatal ultrasound showed that the fetus had enlargement and enhanced echo of the kidneys, in addition with many small renal cysts. Whole exome sequencing showed that the fetus carried pathogenic compound heterozygous variants of the ETFDH gene, namely c.3G>C and c.1436dupA. Sanger sequencing of the family suggested that the variants were inherited from its mother and father, respectively.@*CONCLUSION@#By combining its clinical manifestations and results of whole exome sequencing, the fetus was diagnosed as glutaric acidemia type ⅡC due to the compound heterozygous variants of the ETFDH gene. Above results have provided a basis for prenatal diagnosis and genetic counseling. Fetal exome sequencing has provided an important tool for prenatal diagnosis.

Cyanobacteria based photosynthetic production of sucrose: development and prospect / 生物工程学报

Biorefinery technologies provide promising solutions to achieve sustainable development facing energy and environment crisis, while abundant sugar feedstock is an essential basis for biorefinery industries. Photosynthetic production of sucrose with cyanobacteria is an alternative sugar feedstock supply route with great potentials. Driven by solar energy, cyanobacteria photosynthetic cell factory could directly convert carbon dioxide and water into sucrose, and such a process could simultaneously reduce carbon emissions and supply sugar feedstocks. Here we introduced the history and updated the state-of-the-art on development of cyanobacteria cell factories for photosynthetic production of sucrose, summarized the progress and problems on mechanisms of sucrose synthesis, metabolic engineering strategies and technology expansions, and finally forecasted the future development direction in this area.

Progress and perspectives on cyanobacterial ploidy / 生物工程学报

Cyanobacteria are a phylum of bacteria which are believed to be the oldest photosynthetic prokaryotic microorganisms on earth. The phylogenetic group of cyanobacteria was thought to be one of the prokaryotes that contain monoploid, oligoploid and polyploid species, and one obstacle to engineering cyanobacteria is their polyploidy genome. In recent years, the ploidy level of cyanobacteria was found to be influenced by growth phase and by multiple genetic and environmental factors. In the present article, we reviewed the progress, analytical methods and influencing factors on the cyanobacterial ploidy, and discussed the significance of cyanobacterial polyploidy regarding to environmental ecology and biotechnology. Based on this observation, the future research directions in this field are prospected.

Cyanobacteria cell factories for ethanol photosynthetic production: development and prospect / 生物工程学报

Bioethanol is one of the most promising and representative biofuel products. Photosynthetic production of ethanol using CO₂ and solar energy based on cyanobacteria is of great significance for research and application, due to the potential to reduce CO₂ emission and to provide renewable energy simultaneously. Here we review the history and updated development of cyanobacteria cell factories for ethanol photosynthetic production, the progress and problems in pathway optimization, chassis selection, and metabolic engineering strategies, and finally indicate the future development in this area.

Effects of ggpS over-expression on glycosylglycerol and glycerol biosynthesis of Synechocystis sp. PCC 6803 / 生物工程学报

To study the roles of glucosylglycerol phosphate synthase (Ggps) in glucosylglycerol (GG) and glycerol biosynthesis, we over-expressed Ggps from either Synechocystis sp. PCC 6803 or Synechococcus sp. PCC 7002 in a Synechocystis strain with a high GG titer, and determined the GG and glycerol accumulation in the resultant mutants grown under different NaCl-stress conditions. Ion chromatography results revealed that GG yield was not improved, but glycerol production was significantly enhanced by over-expression of Ggps from Synechocystis sp. PCC 6803 (6803ggpS). In addition, increasing the NaCl concentration of medium from 600 to 900 mmol/L led to a further 75% increase of glycerol accumulation in the mutant strain with 6803ggpS over-expression. These findings show the role of ggpS in driving the carbon flux to the glycerol biosynthesis pathway, and will be helpful for further improvement of GG and glycerol production in Synechocystis.

Structural insights into the catalytic mechanism of aldehyde-deformylating oxygenases

The fatty alk(a/e)ne biosynthesis pathway found in cyanobacteria gained tremendous attention in recent years as a promising alternative approach for biofuel production. Cyanobacterial aldehyde-deformylating oxygenase (cADO), which catalyzes the conversion of Cn fatty aldehyde to its corresponding Cn-1 alk(a/e)ne, is a key enzyme in that pathway. Due to its low activity, alk(a/e)ne production by cADO is an inefficient process. Previous biochemical and structural investigations of cADO have provided some information on its catalytic reaction. However, the details of its catalytic processes remain unclear. Here we report five crystal structures of cADO from the Synechococcus elongates strain PCC7942 in both its iron-free and iron-bound forms, representing different states during its catalytic process. Structural comparisons and functional enzyme assays indicate that Glu144, one of the iron-coordinating residues, plays a vital role in the catalytic reaction of cADO. Moreover, the helix where Glu144 resides exhibits two distinct conformations that correlates with the different binding states of the di-iron center in cADO structures. Therefore, our results provide a structural explanation for the highly labile feature of cADO di-iron center, which we proposed to be related to its low enzymatic activity. On the basis of our structural and biochemical data, a possible catalytic process of cADO was proposed, which could aid the design of cADO with improved activity.

Effects of captopril in treatment of acute pancreatitis and its influence on serum C reactive protein / 中国基层医药

Objective To evaluate the effect of captopril in treatment of acute pancreatitis and its influence on serum C reactive protein ( CRP) .Methods 100 cases of acute pancreatitis patients were randomly divided into the control group and observation group,50 cases in each group.The control group was treated with fasting,gastroin-testinal decompression,somatostatin, anti-inflammatory and other conventional treatment, while the observation group was given captopril on basis of routine treatment,the effective rate after 7 days of treatment,pain relief time,amylase returned to normal time,pancreatic edema improved time,serum CRP before and after treatment were compared be-tween the two groups of patients.Results The effective rate after 7 days of treatment of the observation group was 86%,which was significantly higher than 72%of the control group(χ2 =7.334,P<0.05);The pain relief time,am-ylase returned to normal time,pancreatic edema improved time of the observation group after treatment were (1.75 ± 0.44)d,(3.49 ±1.22)d,(7.50 ±2.39)d,respectively,which of the control group were (2.25 ±0.58)d,(4.25 ± 1.59)d,(9.25 ±2.87)d,respectively,there were statistically significant difference between the two groups(t1 =3.55,t2 =4.18,t3 =8.33,all P<0.05);the serum CRP level of the observation group after treatment was (10.39 ± 3.66)mg/L,improved obviously compared to (25.46 ±8.33)mg/L of the control group,the differences between the two groups was statistically significant(t=11.45,P<0.05).Conclusion Captopril has a good effect in treatment of acute pancreatitis,which can effectively improve the level of serum CRP.

Construction and evaluation of efficient gene expression platforms in Synechocystis sp. strain PCC6803 / 生物工程学报

For metabolic engineering of cyanobacteria, there is an urgent need to construct a group of efficient heterologous gene expression platforms and to evaluate their expression efficiencies. Here we constructed three integrative vectors, the pKW1188-derived pFQ9F, pFQ9R and pFQ20, for integration of heterologous genes into the genome of the model cyanobacteria strain Synechocystis sp. strain PCC6803. The pFQ16, an RSF1010-derived broad host range shuttle vector, was constructed for conjugative transfer of genes to various cyanobacteria strains. All the four platforms constructed here applied the rbc (encodes Ribulose-1, 5-bisphosphate carboxylase/oxygenase) and the rbc terminator to promote and terminate the gene transcription. Besides, a "Shine-Dalgarno -AUG" fusion translation strategy was used to keep the high protein translation efficiency. Using lacZ as a reporter gene, the expression efficiency of pFQ20 was evaluated and showed a strong beta-galactosidase expression (109 Miller). Furthermore, the platform pFQ20 was used to express the E. coli tesA' gene and showed significant protein bands through the Western Blot test. The expression platforms constructed in this study offer useful molecular tools for metabolic engineering of cyanobacteria in the future.

Improvement of fatty acid ethyl ester production by optimizing thioesterase expression / 生物工程学报

Biosynthesis of fatty acid ethyl ester (FAEE) by genetically engineered Escherichia coli has attracted extensive attentions from scientific community. In this study, we evaluated the effects of thioesterase with different origins on FAEE production and the results show that Cc FatB1 from Cinnamomum camphorum is better than tesA' from E. coli for FAEE production. Then, the optimized FAEE-producing strain KC4, with 21.4 mg/(L x OD600) FAEE production under flask condition and 31.16 mg/(L x OD600) under 5 L fermentation condition, was constructed by co-expression of Cc FatB1 and tesA'. Compared with the reported FAEE-producing strain KC3, KC4 possesses the higher FAEE productivity.

Value of endoscopic ultrasound in distinguishing ectopic pancrease and stromal tumor in stomach / 中华超声影像学杂志

Objective To explore the value of endoscopic ultrasound (EUS) in differential diagnosis of ectopic pancreas and stromal tumor in stomach.Methods The clinical data and endoscopic ultrasound findings of 40 patients with ectopic pancrea and 141 patients with stromal tumor were retrospectively analyzed,including the location,size,growth pattern,layer of origin,internal echo pattern and so on.All patients were diagnosed pathologically.Results Ectopic pancreas were predominantly located in the antrum (92.5％) and stromal tumor were mainly located in the gastric body (45.4％),the antrum (23.4％) and the fundus (22.0％),there was a significant difference in lesion' s location between two diseases (P ＜0.001).There was no statistical difference in the ratio of longest/shortest diameter of the lesion size(P =0.057).But there was statistical difference in lesion size between ectopic pancrease and stromal tumor [(19.98 ± 12.80)mm vs (11.25 ± 3.61)mm].Mural growth pattern was most common in ectopic pancrease (82.5％) and mural growth pattern (37.6％),extraluminal growth pattern (24.8％) and intraluminal growth pattern (22.7％) in stromal tumor (P ＜0.001).The third layer (75.0％) was predominant in ectopic pancrease while the fourth (54.6％) and the second layers (34.0％) in stromal tumor (P ＜0.001).With regard to internal echo pattern,hypoechoic (32.5％),intermediate (27.5％) and heterogeneous (27.5％) was predominant in ectopic pancrease and homogeneous hypoechoic lesion (72.3％) in the stromal tumor (P ＜ 0.001).Conclusions There were statistical differences between ectopic pancrease and stromal tumor in the location,size,layer of origin,growth pattern and internal echo pattern under EUS.EUS plays an important role in identification of ectopic pancrease and stromal tumor.

Characterization of a key gene in membrane lipid cycle in Synechocystis sp. PCC6803 / 生物工程学报

Free fatty acid profiles of wild type and fatty acyl-ACP synthase deletion mutant strain of Synechocystis sp. PCC6803 indicated that one origin of these fatty acids is the process of lipid remodeling or lipid degradation. Lipase is the key enzyme involved in this process. The gene sll1969 is the sole gene encodes a putative lipase in Synechocystis sp. PCC6803. To identify the function of this gene and its role in fatty acid metabolism, we cloned the sll1969 from genomic DNA, overexpressed it in Escherichia coli BL21 (DE3) using pET expression system and purified this recombinant enzyme with Nickel-nitrilotriacetic acid affinity chromatography. The enzyme activity was assayed by spectrophotometric with p-nitro-phenylbutyrate as substrate. The K(m) and k(cat) of the enzyme is (1.16 +/- 0.01) mmol/L and (332.8 +/- 10.0)/min, respectively toward p-nitro-phenylbutyrate at 30 degrees C. The optimal temperature of the enzyme is 55 degrees C. To investigate the biological role of Sll1969 in fatty acid metabolism in cyanobacteria, we constructed sll1969 deletion and overexpression mutant strains in the background of fatty acyl-ACP synthase deletion mutant of Synechocystis sp. PCC6803. The analyses of the content of free fatty acids in different mutant strains showed that the contents of Sll1969 and free fatty acid are positively correlated. The free fatty acid profiles of the sll1969 mutant strains suggested this enzyme is not the sole enzyme for degrading lipid in Synechocystis sp. PCC6803.

Study on species identification of brucella with small fragments of PCR / 中华检验医学杂志

Objective To identify the species of brucella rapidly with small fragments of PCR products. Methods The primers were designed according to Brucella spp. repeated insertion sequence IS711 in addition to the specific gene sequences of Brucella melitensis, Brucella abortus and Brucella suis. The small fragments of 3 standard strains and 13 clinical isolates of Brucella were amplified by PCR. Then PCR products were T-A cloned and sequenced. The DNA of standard strains were diluted for sensitivity and stability testing. Results Four specific PCR products were obtained by PCR (63, 67, 81 and 83 bp). The results of T-A cloning and sequencing were in accord with the target gene fragment test. The detection limit of DNA was 1 μg/L. The expected results were achieved with different primers. Conclusion Species identification of Brucella with small fragments of PCR production is a method to identify Brucella strains with great specificity,sensitivity and stability.

Relationship among mTOR and HIF-1? expression and angiogenesis in gastric cancer / 中国现代普通外科进展

Objectives: To detect the expressions of mTOR and HIF-1?and their relationship with angiogenesis in gastric carcinomas. Methods: The expressions of mTOR and HIF-1?in gastric carcinomas tissues(n=70)and normal tissues(n=45)were measured by immunohistochemistry staining.CD34 immunostaining was used to measure the microvascular density(MVD).Results: The rates of the mTOR and HIF-1? positive expression were 51.4% and 67.1% respectively. MVD value of gastric cancer tissues was much higher than that of normal tissues(t=4.358, P

Study on the connection between adiponectin and chronic hepatitis B associated with liver fibrosis / 中国医师进修杂志

Objective To explore the adiponectin expression in the chronic hepatitis B (CHB) patients and compare the level of edipanectin before and after antiviral therapy, and investigate the relationship between ediponectin and CHB. Methods Serum adiponectin was detected by enzyme linked immunosorbent assay (ELISA), and the expression of adipanectin in liver tissue was detected by palymerase chain reaction (PCR) and immunohistochemistry. The liver functions and HBV DNA were also tested. Liver biopsy tissue was assessed by fibrosis stage. Results Serum adipanectin in CHB liver cirrhosis [(20.65 ± 1.29) mg/L]was higher than that in other stage fibrosis, the values were (8.57 ± 0.28 ), (12.22 ± 0.64), (15.22 ± 0.77 ) mg/L (P< 0.01 ). Serum adiponectin was positively associated with fibrosis stage(r = 0.976,P< 0.01 ). There was no significant difference between HBV DNA and ALT (P> 0.05 ). Serum adipanectiu was independently associated with liver cirrhosis (r = 1.07, P = 0.02, 95% CI = 1.00-l.14).At the end of antiviral therapy,the expressions of adiponectin declined in patients with improved fibrosis and sustained viral response. Conclusion Serum adiponectin may play a role in CHB fibrosis, and reduced adipanectin level is associated with improved fibrosis after antivital therapy.

Role of IL-17A and Th17 in inflammatory bowl disease / 中国现代普通外科进展

Objective:To detect the expression of IL-17A、IL-10 and IFN-? in patients wth different forms of inflammatory bowel disease(IBD) during clinical remission phase and investigate their role in the onset and treatment of IBD.Methods:Tissue samples were obtained from patients with ulcerative colitis(UC,n=46),Crohn's disease(CD,n=12),and normal colorectal tissues(n=20).IL-17A expression was evaluated by a standard immunohistochemical procedure.Serum IL-17A、 IL-10 and IFN-? levels were determined by ELISA.Results:IL-17A expression was not detected in samples from normal colonic mucosa but was present in the mucosa of IBD.The level of IL-17A significantly increased in IBD patients while it was not detected in the sera of normal individuals.The level of IL-10 in patients with Crohn's disease was significantly higher than that in patients with ulcerative colitis and control group(P

Slit2/Robo1 signaling expression and its correlation with angiogenesis in gastric cancers / 中国现代普通外科进展

Objective: To investigate the expressions of Slit2 and its receptor Robo1 in human gastric carcinomas. Methods: The expression of Slit2 protein, Robo1 protein and CD34-labeled microvessel density(MVD) were measured by immunohistochemical staining (SP) in 54 cases of gastric carcinomas and 28 cases of Para-cancer tissues. Results:The positive rates of Slit2 ,Robo1 were 63.0% and 77.8% and the expression of Slit2 , Robo1 and MVD in cancerous tissues were higher than those in para-cancer tissues2(?2=26.586,P