Advances of development and application amino acid biosensors / 生物工程学报

Amino acids are the basic building blocks of protein that are very important to the nutrition and health of humans and animals, and widely used in feed, food, medicine and daily chemicals. At present, amino acids are mainly produced from renewable raw materials by microbial fermentation, forming one of the important pillar industries of biomanufacturing in China. Amino acid-producing strains are mostly developed through random mutagenesis- and metabolic engineering-enabled strain breeding combined with strain screening. One of the key limitations to further improvement of production level is the lack of efficient, rapid, and accurate strain screening methods. Therefore, the development of high-throughput screening methods for amino acid strains is very important for the mining of key functional elements and the creation and screening of hyper-producing strains. This paper reviews the design of amino acid biosensors and their applications in the high-throughput evolution and screening of functional elements and hyper-producing strains, and the dynamic regulation of metabolic pathways. The challenges of existing amino acid biosensors and strategies for biosensor optimization are discussed. Finally, the importance of developing biosensors for amino acid derivatives is prospected.

Effect of Shaoyaotang on Expressions of TLR4, NF-κB p65 and IL-6 in Rats with Damp-heat Ulcerative Colitis / 中国实验方剂学杂志

Objective:To explore the mechanism of Shaoyaotang in the treatment of ulcerative colitis (UC) based on toll-like receptor 4 (TLR4)/nuclear factor kappaB (NF-κB) signaling pathway. Method:A total of 50 Wistar rats were selected, including half male and half female. The damp-heat UC rat model was replicated by the methods of the combination of diseases and syndromes and the combination of 2, 4, 6-nitrobenzene sulfonic acid (TNBS) and ethanol. After the successful modeling, the model rats were randomly divided into model group, salazulesulfonate group, and low, medium and high-dose Shaoyaotang groups, and 10 rats (half male and half female) were selected as the blank control group. Low, medium and high-dose Shaoyaotang groups were given 6, 12, 24 g·kg-1 by gavage, and salazonyl arsenic group was given 1 g·kg-1 by gavage. Blank control group was given the equal volume of normal saline for 21 consecutive days. Colon samples were collected after the last administration, and the expressions of TLR4, NF-κB p65 and IL-6 mRNA in colon tissues were detected by fluorescent quantitative polymerase chain reaction (Real-time PCR）, and the expressions of TLR4, NF-κB p65 and IL-6 protein in colon tissues were detected by Western blot. Result:Compared with the blank control group, the relative expressions of TLR4, NF-κB p65, IL-6 mRNA and protein in the model group were significantly increased (P<0.05). Compared with the model group, the expression levels of TLR4, NF-κB p65 and IL-6 mRNA and protein in the salazopyridine group and Shaoyaotang groups were significantly decreased (P<0.05). Conclusion:Shaoyaotang can inhibit the development of UC by regulating the expressions of TLR4, NF-κB p65 and IL-6 mRNA and proteins in the TLR4/NF-κB pathway.

Ginsenoside Rb1 improving autophagic flux against myocardial ischemia reperfusion injury in isolated-heart of rat / 解剖学报

Objective To explore the protective effect of ginsenoside Rb1 on the myocardial ischemia / reperfusion (I / R) injury in rats in vitro. Methods Totally 60 adult male SD rats were randomly divided into 6 groups:sham group,I / R group,ginsenosde Rb1 pretreatment groups(at the doses of 1 μmol / L,5 μmol / L,10 μmol / L and 20 μmol / L,respectively), 10 in each group. The Langendorff perfusion system was used to establish I / R model. The Lab Chart electrophysiological system was used to monitor real-time heart function by monitoring heart rate (HR), left ventricular development pressure (LVDP) and left ventricular development pressure (± dp / dtmax). TTC staining method was used to measure myocardial infarct size. The Western blotting were used to assay Beclin 1, LC3, p62 and Lamp 2 expression, respectively. The immunohistochemistry were used to assay Beclin 1 expression. Results Ginsenoside Rbl of all the four different concent rations improved the decrease of LVDP and ± dp / dtmax arising from myocardial I / R injury. Meanwhile, ginsenoside Rbl significantly decreased the area of cardial infarction. Ginsenoside Rb1 (10 μmol / L) precondition group protected the heart most significantly (P<0. 05). The expression of Beclin 1 with I / R increased significantly in the cytoplasm of cardiomyocytes. Moreover, Beclin 1 expression decreased after addition pretreatment with ginsenoside Rb1 (10 μmol / L) (P < 0. 05). Compared with sham group, we found that the autophagic flux was impaired in I / R group which the expression of Beclin 1, LC3 and p62 increased significantly, as well as the expression of Lamp 2 decreased significantly. On the other hand, pretreatment with ginsenoside Rb1 (10 μmol / L) could reverse impaired autophagic flux (P < 0. 05). Conclusion Ginsenoside Rbl demonstrates pharmacological preconditioning effect and protects against myocardial I / R injury by improving damaged-autophagy flux, the dose of 10 μmol / L precondition protectes the heart most significantly.

Effect of Shaoyaotang on Expressions of AP-1 and TNF-α in Colon of Rats with Hygrothermal Ulcerative Colitis / 中国实验方剂学杂志

Objective: To observe the effect of Shaoyaotang on mRNA and protein expressions of colon tissue activated protein-1 (AP-1) and tumor necrosis factor-α (TNF-α) of hot and humid-type intrinsic ulcerative colitis (UC) model in rats, in order to explore the mechanism of action of herbaceous peony decoction in the treatment of UC. Method: Totally 60 Wistar rats were randomly divided into blank group, model group, SASP group, and low, medium and high-dose Shaoyaotang groups. The damp-heat intrinsic UC rat model was replicated based on integrated disease and syndrome, namely, high-fat and high-sugar spicy food and immune complex method combined with 2,4,6-trinitrobenzene sulfolnic acid (TNBS) and ethanol complex method. After the successful modeling, low, medium and high-dose Shaoyaotang (6, 12, 24 g·kg-1) was given by gavage, and 1 g·kg-1 dose of salazol sulfadiazine was given to by gavage. The blank group was given constant volume normal saline for 21 d. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was used to detect mRNA expressions of AP-1 and TNF-α in colon tissues, and Western blot was used to detect protein expressions of AP-1 and TNF-α in colon tissues. Result: Compared with the blank group, relative mRNA and protein expressions of AP-1, TNF-α in the model group were significantly increased (Pα in the treatment groups were significantly decreased (PConclusion: Shaoyaotang can inhibit the expression of TNF-α and stimulate AP-1 protein expression in rats with damp-heat UC.

Expression changes of cell cycle related molecules in palatal tissue of fetal mice with cleft palate induced by TCDD / 中华整形外科杂志

Objective@#The purpose of this study is to investigate the expression change of cell cycle-related molecules in platal tissue of fetal mice with cleft palate, induced by 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD), and to explore the mechanism of cell cycle-related molecules in cleft palate.@*Methods@#In vivo, 48 pregnant mice were randomly divided into TCDD treatment group and control group with Random number table, 24 mice in each group. On the embryonic day 10.5 (E10.5), pregnant mice were orally administrated with TCDD 28 μg/kg (containing 5 μg/ml TCDD of corn oil) in TCDD treatment group. The same volume of corn oil was given to the mice in control group. The pregnant mice in each group were sacrificed on E13.5, E14.5 and E15.5, to collect the fetal palates for analysis. Fetal palates were used to extract total RNA and total protein, so as to detect the expression levels of cell cycle-related molecules, using RT-PCR and western blotting respectively. In vitro, human kidney embryo 293t (HEK293t) cells were treated with different concentrations of TCDD (0.01, 0.1, 0.5 and 1 nmol/L), and cells proliferation activity was detected using MTT assay. Statistical analysis was performed with IBM SPSS 24.0. Kolmogorov-Smimov test was used for normal distribution check, and the distribution was normal. Independent t-test was carried out among two groups. P<0.05 was considered statistically significant.@*Results@#At E13.5, E14.5 and E15.5, the expression level of interferon regulatory factor 6 (Irf6) protein were higher in the control group (1.26 ± 0.13, 1.67 ± 0.14 and 1.42 ± 0.15, respectively) compared to that in the TCDD group (0.81 ± 0.08, 1.04± 0.02 and 0.86 ± 0.12, respectively), on each time point (t value were 2.836, 3.662 and 2.867, respectively; P values were 0.0471, 0.0146 and 0.0241, respectively). The expression level of cyclin-dependent kinase inhibitor 1A (P21) protein on E13.5 and E14.5 of the control group (2.26 ± 0.21, 1.99 ± 0.21)were higher than that in the TCDD group on each time point(1.43 ± 0.12、0.93 ± 0.22), (t value were 3.398 and 3.378; P value were 0.8726 and 0.0273). The expression level of cyclin D1 in the control group (1.00±0.02, 0.94±0.03 and 1.11±0.09, respectively)were higher than that of the TCDD group (0.28±0.01, 0.33±0.06 and 0.88±0.01, respectively) on each time point (t value are were 22.53, 22.35 and 14.27, respectively, P value <0.001, <0.001 and<0.001, respectively). The expression of cyclin E1, cyclin A2, cyclin B1, CDK6, CDK2 and CDK1 in TCDD groups were higher than that of the controls (P<0.05). However, there was no significant difference of cyclin B1 on E13.5 and Cdk2 on E15.5. As treatment with TCDD (0.1 nmol/L) at 1, 2 and 3 days (0.70 ± 0.05, 1.05 ± 0.03 and 1.39 ± 0.04, respectively), the proliferation of HEK293t cells increased compared with the control group (0.49 ± 0.04, 0.98 ± 0.03 and 1.55 ± 0.02, respectively). The differences were statistically significant (t value were 2.829, 1.395 and 2.692, respectively; P value were 0.0198, 0.1320 and 0.0247, respectively).@*Conclusions@#TCDD down-regulates Irf6 and P21, and interferes with the normal expression of cell cycle-associated molecules, which in turn interferes with medial edge epithelia (MEE) cells cycle arrest and proliferation. These indicate that the disorder of spatiotemporal expression of cell cycle-related molecules during palatal development may be involved with the mechanism of TCDD-induced cleft palate..

1H-MRS detection of metabolites in posterior cingulate gyrus of Parkinson disease with cognitive impairment patients / 中国医学影像技术

Objective To investigate the value of 1H-MRS technology combined with linear combination model (LCmodel) software in diagnosis of Parkinson disease (PD) cognitive impairment.Methods Thirty-five PD patients (PD group) and 22 matched healthy subjects (control group) were collected.Patients in PD group were divided into PDN and PDMCI subgroups according to whether having cognitive impairment or not.The concentration of metabolites of posterior cingulate gyrus (PCG)was applied with 1H-MRS technology combined with LCmodel software.The differences of metabolites were compared between the two groups,and the correlations between metabolites level and cognitive status were analyzed.Results The absolute concentrations of metabolites in PDN subgroup were not significantly different from those in control group (all P＞0.05).The absolute concentrations of total creatine (tCr),N-acetyl aspartate (NAA),myo-inositol (mI) and glycerophosphocholine+ phosphocholine (tCho) in PDMCI subgroup were lower than those in control group (all P＜0.05).The absolute concentration of tCr in PDMCI subgroup was lower than that in PDN subgroup (P＜0.05).There was positive correlation among the absolute concentration of tCr (r=0.444,P=0.01),glutathione (GSH;r=0.393,P=0.024) and MMSE scores,as well as among the absolute concentration of tCr (r=0.367,P=0.035),GSH (r=0.376,P=0.031),tCho (r=0.375,P=0.031) and MoCA scores.Conclusion 1 H-MRS technology combined with LCmodel software can quantitatively analyze the changes of metabolites in PCG,therefore being helpful to evaluating PD cognitive impairment.

Based on the teeth in vitro skull specimen to investigate CARE kV CT scan technology of decreasing radiation dose / 实用医学杂志

Objective Based on the teeth in vitro skull specimen to investigate the feasibility and accuracy of CARE kV CT scan for reducing radiation dose. Methods Of eight fixed and preserved skull teeth were scanned repeatedly in groups of A, B, C. In expose factors of 100 kVp/150 mAs in conventional scanning of group A, open CARE Dose 4D scan of group B and CARE kV scan of group C. We assessed the radiation dose in the group A, B and C, the average CT values of the images, noise, SNR, CNR and subjective ratings. Results Three groups of actual scan tube current, tube voltage and radiation dose were statistically significant (P 0.05). Conclusion CARE kV technology based on the use specimens tooth can reduce the radiation dose, protect image quality, and be consistent to the reports of previous researches.

Application of low kilovoltage combined with sinogram affirmed iterative reconstruction in assessment of calcified plaque in coronary artery / 实用医学杂志

Objective To investigate the performance of dual source CT scanning combined with low-tube-voltage iterative reconstruction (SAFIRE) for demonstrating coronary artery calcium plaques. Methods 70 patients with clinically suspected coronary atherosclerosis and with normal BMI received SAFIRE-3 or filtered back projection (FBP) for reconstruction after undergoing CT scans with tube voltage range of 100 kV. The CT images (average CT values, noise, SNR, and CNR) and subjective scores (number of calcium plaque, calcium plaque adjacent artifacts, and image quality) were compared between the two groups. Results The average CT value of the aortic root did not differ significantly between the two procedures for reconstruction (P > 0.05). Ob jective evaluation of the images (noise, SNR, and CNR) between the two groups was statistically significant (P < 0.05 for all comparisons), so was image subjective rating (number of calcium plaque and rate of calcium plaque edge)(P < 0.05 for all comparisons). SAFIRE-3 reconstruction was markedly superior to FBP reconstruc-tion. Conclusions Low kilovoltage combined with reconstruction of SAFIRE-3 can reduce calcium plaque adja-cent artifacts and enhance quality of images , which is better than FBP in displaying the morphology and adjacent conditions of calcified plaque; therefore it is worth popularizing.

Laparoscopic permanent sigmoid stoma creation through the extraperitoneal route versus transperitoneal route, [A meta-analysis of stoma-related complications]

To compare laparoscopic extraperitoneal colostomy with transperitoneal colostomy for construction of a permanent stoma by measuring the incidence of parastomal hernia, and other postoperative complications related to colostomy. The meta-analysis was carried out in the General Surgery Department of the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China in 2014. A literature search of Medline, EMBASE, Cochrane database, and the Chinese Biomedical Literature Database [CBM] from the years 1990 to 2014 was performed. The literature searches were carried out using medical subject headings and free-text words: extraperitoneal colostomy, transperitoneal colostomy, laparoscopic extraperitoneal colostomy, rectal cancer, laparoscopic abdominoperineal resection, parastomal hernia, permanent stoma, and colostomy-related complications. Two different reviewers carried out the search and evaluated studies independently. One randomized controlled trial and 6 retrospective studies were included. A total of 378 patients [209 extraperitoneal colostomy and 169 transperitoneal colostomy] were identified. Our analysis showed that there was a significantly lower rate of parastomal hernia [odds ratio 0.10; 95% confidence interval 0.03-0.29, p<0.0001] in the extraperitoneal colostomy group. However, the other stoma-related complications were not significantly different between the 2 groups. Colostomy construction via the extraperitoneal route using a laparoscopic approach can largely reduce the incidence of parastomal hernia. Laparoscopic permanent sigmoid stoma creation through the extraperitoneal route should be the first choice after laparoscopic abdominoperineal resection

Clinical and MRI features of posterior reversible encephalopathy syndrome in patients with systemic lupus erythematosus / 中华神经科杂志

Objective To investigate the clinical and MRI features and pathogenic mechanism of posterior reversible encephalopathy syndrome (PRES) in patients with systemic lupus erythematosus (SLE).Methods Six cases of PRES in SLE proved by integrated clinical diagnosis were collected from January 2008 to December 2013.The clinical and MRI features of these patients were studied retrospectively and the related literatures were reviewed.Results The initial episode of nervous system was involved in 4 cases.The clinical presentations were headache (2 cases),seizures (4 cases),acute confusion state (2 cases),altered mentation (1 case) and vision change (1 case).All cases had lupus nephritis and hypertension,in which 4 cases had renal failure;6 cases were treated with immunosuppressive agents and 4 cases with cyclophosphamide for lupus nephritis when they developed PRES.The vasogenic edema lesions were distributed in the parietal or occipital lobe (5 cases),the frontal lobe (4 cases),temporal lobe (4 cases),basal ganglia (3 cases),splenium (1 case) and cerebellar hemispheres (2 cases).Three major patterns of PRES included dominant parietal-occipital (2 cases),the holohemispheric watershed (3 cases),and superior frontal sulcal (1 case).Conclusions The clinical and MRI findings of PRES in patients with SLE are typical.Awareness of this entity as early as possible can help to improve curative effect and prognosis.

Comparsion on therapeutic effect of hydroxyl - glucoside and sodium hyaluronate on dry eye after age-related cataract surgery / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the tear film changes after phacoemulsification combined with intraocular lens implantation in age - related cataract patients and to compare the therapeutic effect of hydroxyl-glucoside and sodium hyaluronate on the postoperative dry eye. METHODS:A total of 49 patients ( 70 eyes ) with age-related cataract suffering from dry eye after phacoemulsification combined with IOL implantation were divided into treatment group 1 ( group A, 23 eyes with conventional therapy and hydroxyl - glucoside at 7d postoperatively) , treatment group 2 ( group B, 22 eyes with conventional therapy and sodium hyaluronate at 7d postoperatively) and control group ( group C, 25 eyes with conventional therapy only ) . Questionnaire score of dry eye symptoms, SchirmerⅠtest ( SⅠt) , tearfilm break up time ( BUT ) and corneal fluoresce in staining ( CFS ) were measured at 2d preoperatively and 7, 14, 30, 90d postoperatively. RESULTS: No statistical differences existed among the three groups of preoperative 2d ( P > 0. 05 ). At 2d preoperatively and 90d postoperatively, the results of questionnaire score of dry eye symptoms, SⅠt, BUT, and CFS displayed no statistical differences in the patients of three groups (P>0. 05). While there was statistical significance among preoperatively and 7, 14, 30d postoperatively of the three groups (PCONCLUSION: At the early stage after phacoemulsification combined with IOL implantation, the tear film stability is decreased, which may promote eyesymptoms. Management with hydroxyl-glucoside or sodium hyaluronate plays a role in relief of the structure and stability of the tear film and improves dry eye symptoms, while sodium hyaluronate eye drops is more effective.

Assessment of a five-color flow cytometric assay for verifying automated white blood cell differentials / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>White blood cell (WBC) counts and differentials performed using an automated cell counter typically require manual microscopic review. However, this last step is time consuming and requires experienced personnel. We evaluated the clinical efficiency of using flow cytometry (FCM) employing a six-antibody/five-color reagent for verifying automated WBC differentials.</p><p><b>METHODS</b>A total of 56 apparently healthy samples were assessed using a five-color flow cytometer to verify the normal reference ranges of WBC differentials. WBC differentials of 622 samples were also determined using both a cell counter and FCM. These results were then confirmed using manual microscopic methods.</p><p><b>RESULTS</b>The probabilities for all of the parameters of WBC differentials exceeded the corresponding normal reference ranges by no more than 7.5%. The resulting WBC differentials were well correlated between FCM and the cell counter (r > 0.88, P < 0.001), except in the case of basophils. Neutrophils, lymphocytes, and eosinophils were well correlated between FCM and standard microscopic cytology assessment (r > 0.80, P < 0.001). The sensitivities of FCM for identification of immature granulocytes and blast cells (72.03% and 22.22%, respectively) were higher than those of the cell counter method (44.92% and 11.11%, respectively). The specificities of FCM were all above 85%, substantially better than those of the cell counter method.</p><p><b>CONCLUSION</b>These five-color FCM assays could be applied to accurately verify abnormal results of automated assessment of WBC differentials.</p>

Proximal gastrectomy versus total gastrectomy for proximal gastric carcinoma. A meta-analysis on postoperative complications, 5-year survival, and recurrence rate

To compare proximal gastrectomy [PG] with total gastrectomy [TG] for proximal gastric carcinoma, through the 5-year survival rate, recurrence rate, postoperative complications, and long-term life quality. The meta-analysis was carried out in the General Surgery Department of the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China. We searched Medline, EMBASE, and the Cochrane Library from June to November 2012. The literature searches were carried out using medical subject headings and free-text word: 'proximal gastrectomy' 'total gastrectomy' 'partial gastrectomy' 'stomach neoplasms' and 'gastric cancer'. Two different reviewers carried out the search and evaluated studies independently. Two randomized controlled trials and 9 retrospective studies were included. A total of 1364 patients were included in our study. Our analysis showed that there is no statistically significant difference in 5-year survival rate between PG and TG [60.9% versus 64.4%]. But, the recurrence is higher in the PG group than the TG [38.7% versus 24.4%]. The anastomotic stenosis rate is also higher in the PG than the TG [27.4% versus 7.4%]. Proximal gastrectomy is an option for upper third gastric cancer in terms of safety. However, it is associated with high risk of reflux symptoms and anastomotic stenosis. Therefore, TG should be the first choice for proximal gastric cancer to prevent reflux symptoms

The establishment and research of ventilator quality control information system / 中国医疗器械杂志

Ventilator quality control is an important task for the management of hospital medical equipment. Analysis the problems of hospital medical device quality control and introducing ventilator quality control information system design and technical characteristics. The application of the system can improve the efficiency of quality control, and enhance hospital medical equipment management level.

Deficiency of succinic dehydrogenase or succinyl-coA synthetase enhances the production of 5-aminolevulinic acid in recombinant Escherichia coli / 生物工程学报

5-aminolevulinic acid (ALA), a precursor for biosynthesis of pyrrole compounds in living organisms, has been widely used in agriculture and medical photodynamics therapy and is regarded as a promising value-added bio-based chemical. In the previous investigations on ALA production with recombinant Escherichia coli expressing heterogenous C4 pathway gene, LB media supplemented with glucose and ALA precursors succinate and glycine is widely used, leading to high production cost. Succinate participates in ALA biosynthesis in a form of succinyl-CoA. In this study, genes involved in succinyl-CoA consumption, sdhAB (encoding succinic dehydrogenase) or sucCD (encoding succinyl-CoA synthetase) of E. coli MG1655 was knocked out and tested for ALA accumulation. In comparison with the recombinant E. coli strain expressing heterogenous ALA synthetase, the sdhAB- or sucCD-deficient strain accumulate 25.59% and 12.40%, respectively, more ALA in a 5 L fermentor using a defined synthetic medium with glucose as main carbon source and without supplementation of succinate, providing a novel cost-effective approach for industrial production of ALA.

Fistula plug versus conventional surgical treatment for anal fistulas A system review and meta-analysis

To evaluate the recurrence and fecal incontinence of anal fistula plug versus conventional surgical treatment for anal fistulas. This meta-analysis was carried out in the General Surgery Department of the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China. We searched the Medline, EMBASE, and the Cochrane Library from June 2011 to April 2012. The literature searches were carries out using medical subject headings and free-text word: anal fistula, fibrin adhesive, fibrin sealant, and fistula plug. Two randomized controlled trials and 3 retrospective controlled studies were included. A total of 428 patients were included in our study. The recurrence rate was higher in those patients who accept fistula plug treatment [62.1% versus 47%] [p=0.004]. Anal fistula plug has a moderate probability of success with little risk of incontinence, but the recurrence rate is significantly higher than the conventional surgical treatment. This treatment is minimally invasive, repeatable, and sphinctersparing. This meta-analysis failed to find a statistically significant difference in incontinence rate between conservative treatment and conventional surgical treatment

Software for Hospital Infection Control / 中华医院感染学杂志

OBJECTIVE To develop the nosocomial infection information management subsystem,as a subsystem of the hospital information system,and to enhance the monitoring methods of hospital infections. It can monitor and provide necessary infection information for all administers of the hospitals. METHODS After collecting the data of hospital infections and analyzing the datum structure of the hospital information system,we developed the software of nosocomial infection information management subsystem using PowerBuilder9.0 and Oracle Database. RESULTS The subsystem could monitor infection of all inpatients,collect and analyze the data of infection examination,patients infected,usage of antibiotics and intervening operations,therefore,relieve information administers from collection works and make them focus on analysis,guidance and problem solutions. CONCLUSIONS As a part of the hospital information system,nosocomial infection information management subsystem can improve the effectivity and efficiency of hospital infection administers due to its accuracy,timeliness and wide coverage.