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1.
World J Gastrointest Surg ; 16(5): 1336-1343, 2024 May 27.
Article in English | MEDLINE | ID: mdl-38817276

ABSTRACT

BACKGROUND: Magnetic anchor technique (MAT) has been applied in laparoscopic cholecystectomy and laparoscopic appendectomy, but has not been reported in laparoscopic partial hepatectomy. AIM: To evaluate the feasibility of the MAT in laparoscopic left lateral segment liver resection. METHODS: Retrospective analysis was conducted on the clinical data of eight patients who underwent laparoscopic left lateral segment liver resection assisted by MAT in our department from July 2020 to November 2021. The Y-Z magnetic anchor devices (Y-Z MADs) was independently designed and developed by the author of this paper, which consists of the anchor magnet and magnetic grasping apparatus. Surgical time, intraoperative blood loss, intraoperative accidents, operator experience, postoperative incision pain score, postoperative complications, and other indicators were evaluated and analyzed. RESULTS: All eight patients underwent a MAT-assisted laparoscopic left lateral segment liver resection, including three patients undertaking conventional 5-port and five patients having a transumbilical single-port operation. The mean operation time was 138 ± 34.32 min (range 95-185 min) and the mean intraoperative blood loss was 123 ± 88.60 mL (range 20-300 mL). No adverse events occurred during the operation. The Y-Z MADs showed good workability and maneuverability in both tissue and organ exposure. In particular, the operators did not experience either a "chopstick" or "sword-fight" effect in the single-port laparoscopic operation. CONCLUSION: The results show that the MAT is safe and feasible for laparoscopic left lateral segment liver resection, especially, exhibits its unique abettance for transumbilical single-port laparoscopic left lateral segment liver resection.

3.
J Hazard Mater ; 415: 125756, 2021 08 05.
Article in English | MEDLINE | ID: mdl-34088210

ABSTRACT

Accumulation of p-hydroxybenzoic acid (PHBA) in soil causes autotoxicity stress in cucumber. When the stress is mitigated by PHBA-degrading bacteria, plant metabolites have not been detected. To explore mechanisms underlining the mitigation, plant metabolites have not been combined with rhizospheric microbes, antioxidant and soil enzymes. In this study, a strain P620 of Klebsiella decomposed PHBA to acetyl CoA. Cucumber was sown into soil supplemented with P620 and/or PHBA. After addition with P620, P620 colonization and the enriched bacterial genera were observed in rhizosphere. Compared to PHBA stress alone, the combination of P620 application and PHBA stress improved plant growth, decreased PHBA concentration in soil, and increased the activities of five soil enzymes and eight antioxidant enzymes in leaves. Metabolomic and transcriptomic analysis highlighted that P620 application decreased the intensities of MAG(18:3) isomer 4, MAG(18:3) isomer 2, lysoPC 18:3 (2n isomer), 2'-deoxyadenosine-5'-monophosphate, pyridoxine, and glucarate O-phosphoric acid in PHBA-stressed leaves and down-regulated the expression of genes related to these metabolites. We propose a mechanism that P620 application alters microbial communities in PHBA-contaminated soil. Thus, the application reduces PHBA concentration in soil, activates antioxidant and soil enzymes, and also influences metabolites in leaves by affecting plant transcriptome, mitigating PHBA stress in cucumber.


Subject(s)
Cucumis sativus , Bacteria/genetics , Hydroxybenzoates , Klebsiella oxytoca , Metabolomics , Rhizosphere , Soil , Soil Microbiology , Transcriptome
4.
World J Surg ; 45(10): 3138-3145, 2021 10.
Article in English | MEDLINE | ID: mdl-34160656

ABSTRACT

BACKGROUND: Cholangiojejunostomy (CJ) is a popular operation; however, no specific anastomotic device is available. A novel magnamosis device for CJ was developed in 2017; here, we evaluated the feasibility and safety of the device. METHODS: Between January 2017 and December 2019, 23 patients who underwent CJ using a novel magnamosis device were enrolled. For the CJ: the parent magnet was placed in the proximal duct, and the purse-string suture was tightened over the rod of the parent magnet. The magnamosis device was introduced into the jejunum, and the mandrel penetrated the jejunum at the anastomotic site, before insertion into the rod of the parent magnet. After rotating the knob, the distance between two magnets was shortened enough to achieve coupling. RESULTS: Sixteen patients (69.6%) underwent open CJ, while 7 (30.4%) underwent laparoscopic CJ; 21 patients (91.3%) underwent choledochojejunostomy, and 2 (8.7%) underwent right or left hepatic duct jejunostomy. The mean time for completion of CJ was 9.2±2.5 min; it was significantly shorter for open CJ than for the laparoscopic way (8±1.2 min vs. 11.8±2.5 min, P<0.05). Only one patient (4.3%) suffered bile leakage after operation and was cured by conservative treatment. The magnets were discharged with a postoperative duration of 66.7±47.2 days, with a 100% expulsion rate. After a median follow-up of 15 months, only one patient (4.3%) developed inflammatory anastomotic stricture. CONCLUSION: The novel magnamosis device is a simple, safe, and effective modality for CJ.


Subject(s)
Jejunostomy , Laparoscopy , Anastomosis, Surgical , Choledochostomy , Humans , Magnets
5.
Front Microbiol ; 11: 522986, 2020.
Article in English | MEDLINE | ID: mdl-33193118

ABSTRACT

Continuous-cropping leads to obstacles in crop productivity by the accumulation of p-hydroxybenzoic acid (PHBA) and ferulic acid (FA). In this study, a strain CFA of Pseudomonas was shown to have a higher PHBA- and FA-degrading ability in media and soil and the mechanisms underlying this were explored. Optimal conditions for PHBA and FA degradation by CFA were 0.2 g/l of PHBA and FA, 37°C, and pH 6.56. Using transcriptome analysis, complete pathways that converted PHBA and FA to acetyl coenzyme A were proposed in CFA. When CFA was provided with PHBA and FA, we observed upregulation of genes in the pathways and detected intermediate metabolites including vanillin, vanillic acid, and protocatechuic acid. Moreover, 4-hydroxybenzoate 3-monooxygenase and vanillate O-demethylase were rate-limiting enzymes by gene overexpression. Knockouts of small non-coding RNA (sRNA) genes, including sRNA 11, sRNA 14, sRNA 20, and sRNA 60, improved the degradation of PHBA and FA. When applied to cucumber-planted soil supplemented with PHBA and FA, CFA decreased PHBA and FA in soil. Furthermore, a reduction of superoxide radical, hydrogen peroxide, and malondialdehyde in cucumber was observed by activating superoxide dismutase, catalase, glutathione peroxidase, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, and monodehydroascorbate reductase in seedlings, increasing the reduced glutathione and ascorbate in leaves, and inducing catalase, urease, and phosphatase in the rhizosphere. CFA has potential to mitigate PHBA and FA stresses in cucumber and alleviate continuous-cropping obstacles.

6.
Front Microbiol ; 9: 1262, 2018.
Article in English | MEDLINE | ID: mdl-29963024

ABSTRACT

Ferulic acid (FA) and p-hydroxybenzoic acid (PHBA) are main phenolic compounds accumulated in rhizosphere of continuously cropped cucumber, causing stress in plants. Microbial degradation of a mixture of FA and PHBA is not well understood in soil. We isolated a strain CSY-P13 of Acinetobacter calcoaceticus, inoculated it into soil to protect cucumber from FA and PHBA stress, and explored a mechanism underlying the protection. CSY-P13 effectively degraded a mixture of FA and PHBA in culture solution under conditions of 39.37°C, pH 6.97, and 21.59 g L-1 potassium dihydrogen phosphate, giving rise to 4-vinyl guaiacol, vanillin, vanillic acid, and protocatechuic acid. During FA and PHBA degradation, activities of superoxide dismutase (SOD), catalase, ascorbate peroxidase, and dehydroascorbate reductase in CSY-P13 were induced. Inoculated into cucumber-planted soil containing 220 µg g-1 mixture of FA and PHBA, CSY-P13 degraded FA and PHBA in soil, increased plant height, and decreased malonaldehyde, superoxide radical, and hydrogen peroxide levels in leaves. CSY-P13 also enhanced SOD, guaiacol peroxidase, catalase, glutathione peroxidase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase activities; increased ascorbate and glutathione contents; and elevated transcript levels of copper/zinc SOD, manganese SOD, and catalase in leaves under FA and PHBA. Moreover, CSY-P13 increased phosphatase, catalase, urease, and sucrase activities and changed bacterial richness, diversity, and community composition by high throughput sequencing in cucumber-planted soil supplemented with the mixture of FA and PHBA. So CSY-P13 degrades the mixture of FA and PHBA in soil and mitigates stress from the two phenolic compounds in cucumber by activating antioxidant enzymes, changing soil bacterial community, and inducing soil enzymes.

7.
Plant Physiol Biochem ; 83: 258-66, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25194776

ABSTRACT

Nitric oxide (NO) and auxin (indole-3-acetic acid; IAA) play vital roles in regulating plants tolerance to abiotic stresses. This study showed that both NO and IAA could induce cucumber plants tolerance to sodic alkaline stress, which depended on their roles in regulating reactive oxygen species (ROS) scavenging, antioxidative enzymes activities, Na(+) accumulation and protecting photosystems II (PSII) from damage. In addition, IAA has significant effect on NO accumulation in cucumber root, which could be responsible for IAA-induced sodic alkaline stress tolerance. Further investigation indicated that the function of IAA could be abolished by NO scavenger (cPTIO). On the contrary, IAA transport inhibitor (NPA) showed no significant effects on abolishing the function of NO. Based on these results, it could be concluded that NO is an essential downstream signal for IAA-induced cucumber tolerance to sodic alkaline stress.


Subject(s)
Cucumis sativus/metabolism , Nitric Oxide/metabolism , Photosystem II Protein Complex/metabolism , Signal Transduction , Sodium/metabolism , Stress, Physiological , Indoleacetic Acids/metabolism
8.
World J Gastroenterol ; 20(30): 10637-41, 2014 Aug 14.
Article in English | MEDLINE | ID: mdl-25132786

ABSTRACT

Splenic hamartoma (SH) is a rare benign tumor usually detected accidentally, which is composed of an aberrant mixture of normal splenic elements. Here, we report a case of 54-year-old man who presented with symptomatic multinodular SH and was admitted initially for thrombocytopenia and anemia. Physical examination revealed that the patients had an anemic appearance and palpable spleen, extending 10 cm below the costal margin. Preoperative ultrasound and computed tomography (CT) indicated splenomegaly with multinodular lesions. On enhanced CT scanning, during the arterial phase, the lesions demonstrated inhomogeneous enhancement, and in the portal phase the lesions were more hyperdense than the splenic parenchyma. The images were highly suggestive of a metastatic tumor. Splenectomy was performed 1 wk later. The tumor was eventually diagnosed as SH according to the morphological features and immunohistochemical detection, by which CD34 was positive in lining cells and some spindle cells, vimentin was positive in the tumor, factor-VIII-related antigen was positive multifocally in lining cells, and smooth muscle actin was positive in some spindle cells. Thrombocytopenia and anemia were cured after splenectomy.


Subject(s)
Hamartoma/diagnosis , Splenic Diseases/diagnosis , Splenic Neoplasms/secondary , Anemia/etiology , Biomarkers/analysis , Biopsy , Diagnosis, Differential , Hamartoma/complications , Hamartoma/metabolism , Hamartoma/surgery , Humans , Immunohistochemistry , Male , Middle Aged , Multimodal Imaging , Predictive Value of Tests , Splenectomy , Splenic Diseases/complications , Splenic Diseases/metabolism , Splenic Diseases/surgery , Thrombocytopenia/etiology , Tomography, X-Ray Computed , Treatment Outcome
9.
World J Surg Oncol ; 12: 220, 2014 Jul 17.
Article in English | MEDLINE | ID: mdl-25029913

ABSTRACT

Necrolytic migratory erythma (NME) is an obligatory paraneoplastic syndrome. Here we describe a woman admitted to the dermatology ward with NME which was later found to be associated with glucagonoma, a slow-growing, rare pancreatic neuroendocrine tumor. Even more rarely, the tumor was located in the pancreas head, while most of such lesions are located in the distal pancreas. The diagnosis of this rare tumor requires an elevated serum glucagon level and imaging confirming a pancreatic tumor. After surgical removal of the tumor, the patient's cutaneous and systemic features resolved. It is therefore imperative that clinicians recognize NME early in order to make an accurate diagnosis and to provide treatment for this rare tumor.


Subject(s)
Glucagonoma/diagnosis , Necrolytic Migratory Erythema/diagnosis , Neuroendocrine Tumors/diagnosis , Pancreatic Neoplasms/diagnosis , Adult , Female , Glucagon/metabolism , Glucagonoma/complications , Glucagonoma/surgery , Humans , Necrolytic Migratory Erythema/complications , Necrolytic Migratory Erythema/surgery , Neuroendocrine Tumors/complications , Neuroendocrine Tumors/surgery , Pancreatic Neoplasms/complications , Pancreatic Neoplasms/surgery , Prognosis
10.
World J Surg Oncol ; 10: 66, 2012 Apr 27.
Article in English | MEDLINE | ID: mdl-22540369

ABSTRACT

We present the case of a 66-year-old woman with a huge gastrointestinal stromal tumor of the stomach that traversed her upper abdomen. The predominant abdominal sign was a huge, palpable mass, but there were no other distinctive findings in her physical examination or her routine blood workup, including biochemical markers. It was difficult to judge the origin of the mass upon imaging. Furthermore, radiological findings revealed that the mass had a complex relationship with many major blood vessels. An exploratory laparotomy revealed a huge tumor protruding from the anterior wall of the stomach fundus, on the lesser curvature of the stomach, measuring approximately 21 × 34 × 11 cm in diameter and weighing 5.5 kg. A complete resection was performed and the tumor was characterized on immunohistochemistry as a gastrointestinal stromal tumor of the stomach. Preoperative diagnosis of gastrointestinal stromal tumors can be difficult, and we hope that the presentation of this rare case and literature review will benefit other diagnosing clinicians having similar problems.


Subject(s)
Gastrointestinal Stromal Tumors/diagnosis , Stomach Neoplasms/diagnosis , Aged , Female , Humans
11.
J Plant Physiol ; 168(9): 927-34, 2011 Jun 15.
Article in English | MEDLINE | ID: mdl-21353326

ABSTRACT

To elucidate the physiological mechanism of chilling stress mitigated by cinnamic acid (CA) pretreatment, a cucumber variety (Cucumis sativus cv. Jinchun no. 4) was pretreated with 50 µM CA for 2d and was then cultivated at two temperatures (15/8 and 25/18 °C) for 1d. We investigated whether exogenous CA could protect cucumber plantlets from chilling stress (15/8 °C) and examined whether the protective effect was associated with the regulation of antioxidant enzymes and lipid peroxidation. At 2d, exogenous CA did not influence plant growth, but induced the activities of some antioxidant enzymes, including superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), guaiacol peroxidase (GPX, EC 1.11.1.7), glutathione peroxidase (GSH-Px, EC 1.6.4.2) and ascorbate peroxidase (APX, EC 1.11.1.11) in cucumber leaves, and it also elevated the contents of reduced glutathione (GSH) and ascorbate (AsA). When CA was rinsed and the CA-pretreated seedlings were exposed to different temperatures, the antioxidant activities in leaves at 3d had undergone additional change. Chilling increased the activities of CAT, GSH-PX, APX, GSH and AsA in leaves, but the combination of CA pretreatment and chilling enhanced the antioxidant activities even more. Moreover, chilling inhibited plant growth and increased the contents of malonaldehyde (MDA), superoxide radical (O2⁻) and hydrogen peroxide (H2O2) in cucumber leaves, and the stress resulted in 87.5% of the second leaves being withered. When CA pretreatment was combined with the chilling stress, we observed alleviated growth inhibition and decreased contents of MDA, H2O2 and O2⁻ in comparison to non-pretreated stressed plants, and found that the withered leaves occurred at a rate of 25.0%. We propose that CA pretreatment increases antioxidant enzyme activities in chilling-stressed leaves and decreases lipid peroxidation to some extent, enhancing the tolerance of cucumber leaves to chilling stress.


Subject(s)
Cinnamates/pharmacology , Cold Temperature , Cucumis sativus/drug effects , Plant Leaves/drug effects , Antioxidants/metabolism , Cucumis sativus/enzymology , Cucumis sativus/physiology , Hydrogen Peroxide/metabolism , Lipid Peroxidation , Malondialdehyde/metabolism , Plant Leaves/enzymology , Plant Leaves/physiology , Reactive Oxygen Species/metabolism , Seedlings/drug effects , Seedlings/metabolism
12.
Physiol Plant ; 132(4): 467-78, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18334000

ABSTRACT

Cucumber (Cucumis sativus L.) cv. Xintaimici (a chilling-resistant cultivar) and cv. Jinyan no. 4 (a chilling-sensitive cultivar) were subjected to two temperatures (15/15 and 25/18 degrees C) under low light (100 mumol m(-2) s(-1)) to understand the relationship between ultrastructural changes and the antioxidant abilities caused by low temperature (15/15 degrees C). We also aimed to find indicators for chilling resistance that could be used on a routine basis in breeding programs of greenhouse crops. At the 15/15 degrees C treatment, the membranes of chloroplast, mitochondrion, ER and plasma were not significantly changed in Xintaimici, whereas they were seriously affected in Jinyan no. 4. This result was consistent with the changes of malonaldehyde in chilling-stressed cucumber leaves. The antioxidant activities were changed under low temperature according to cultivar-expected resistance, relating in part to the described ultrastructural changes. The activities of superoxide dismutase (EC 1.15.1.1) and guaiacol peroxidase (EC 1.11.1.7) increased in chilling-stressed leaves of both cultivars, but the two enzymes were not responsible for the difference between cucumber cultivars. At 15/15 degrees C, contents of GSH and activities of glutathione reductase (GR, EC 1.6.4.2) increased more in leaves of Xintaimici than in those of Jinyan no. 4, while catalase (CAT, EC 1.11.1.6) activities decreased less. GSH, GR and CAT were affected by low temperature and cultivars and correlated with the difference in ultrastructure between chilling-stressed cucumber cultivars. We propose that the three antioxidants might be therefore used as biochemical indicators to screen chilling-resistant cucumber cultivars.


Subject(s)
Antioxidants/metabolism , Cold Temperature , Cucumis sativus/metabolism , Light , Plant Leaves/metabolism , Catalase/metabolism , Cucumis sativus/enzymology , Cucumis sativus/ultrastructure , Glutathione/metabolism , Glutathione Reductase/metabolism , Peroxidase/metabolism , Plant Leaves/enzymology , Plant Leaves/ultrastructure , Superoxide Dismutase/metabolism
13.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 32(1): 138-43, 2007 Feb.
Article in Chinese | MEDLINE | ID: mdl-17344604

ABSTRACT

OBJECTIVE: To determine the clinical application of the new classification of adenocarcinoma of esophagogastric junction (AEG). METHODS: The data of cancer of distal esophagus, cancer of cardia, and proximal gastric cancer were reviewed. Clinicopathologic characteristics, surgical modes and survival were analyzed according to Siewert's standards. RESULTS: Among the 203 patients that were up to the standard, 29 had adenocarcinoma of the distal esophagus (Type I), 80 had true carcinoma of cardia (Type II), and 94 had subcardial carcinoma (Type III). The 5-year survival rates of the 3 types of patients after the operation were 34% for Type I, 27.5% for Type II, and 24.5% for Type III (P<0.05). Further analysis of the patients with curative resection suggested there was no significant difference in the 5-year survival rates, with 37.5% for Type I, 34.5% for Type II, and 33.3% for Type III (P>0.05). CONCLUSION: Difference has been found in the clinicopathologic characteristics of the 3 types of adenocarcinoma of the esophagogastric junction. The exact relation of the 3 types is still unknown. The TNM classification, complete tumor resection and the extent of lymph node metastasis are critical for the prognosis of the patients.


Subject(s)
Adenocarcinoma/classification , Esophageal Neoplasms/classification , Esophagogastric Junction/pathology , Adenocarcinoma/mortality , Adenocarcinoma/surgery , China , Esophageal Neoplasms/mortality , Esophageal Neoplasms/surgery , Esophagectomy , Esophagogastric Junction/surgery , Female , Humans , Male , Retrospective Studies , Survival Analysis , Survival Rate
14.
Appl Microbiol Biotechnol ; 73(3): 533-40, 2006 Dec.
Article in English | MEDLINE | ID: mdl-16802152

ABSTRACT

F(3) and F(4) strains of Aspergillus niger were screened from five strains of fungi to produce multienzyme preparations (containing cellulase, hemicellulase, glucoamylase, pectinase, and acidic proteinase) as dietary supplementation. Enzyme activities indicated that 1:4 (F(3) to F(4)) was the optimum mixture proportion, and 0.3% (W/W) was the preferable pitching rate. In bran mash containing 54.5% (W/W) water, F(3) and F(4) could produce the supplementation better when cultured 30 to 36 h at 30 degrees C. Monofactorial and orthogonal experiments were performed to optimize media. Results of the variance and range analysis showed that the optimum medium contained 80 g of bran, 20 g of cottonseed powder, 1 g of (NH(4))(2)SO(4), and 0.1 g of KH(2)PO(4). When F(3) and F(4 )strains were cultured in the optimum medium containing 54.5% (W/W) water, the activity of cellulase, hemicellulase, glucoamylase, pectinase, and acidic proteinase reached 996; 15,863; 13,378; 7,621; and 5,583 U/g, respectively.


Subject(s)
Animal Feed , Aspergillus niger/enzymology , Aspergillus oryzae/enzymology , Enzymes/biosynthesis , Culture Media , Fermentation , Industrial Microbiology , Multivariate Analysis , Species Specificity , Water
15.
Jpn J Clin Oncol ; 36(6): 364-7, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16766566

ABSTRACT

BACKGROUND: There had never been a clear definition of the cancer of cardia before Siewert's classification, which was proposed in 1996 and approved in 1997 at the second International Gastric Cancer Congress in Munich. On the basis of the classification, this study aims to research into the clinicopathological characteristics and surgical modes of adenocarcinoma of the esophagogastric junction in China. METHODS: The study reviewed the data of the distal esophageal cancer, the cancer of cardia and the proximal gastric cancer at the First Hospital of Xi'an Jiaotong University from January 1995 to December 1999. Surgical patients were defined and classified according to Siewert's classification, and 203 patients were up to the classification. Then the study compared and analyzed the clinicopathological characteristics and the survival rates of the three types of the tumor. RESULTS: Among the 203 patients, there were 29 patients with adenocarcinoma of the distal esophagus (Type I); 80 patients with true carcinoma of cardia (Type II); and 94 patients with subcardial carcinoma (Type III). Obvious differences were found in the clinicopathological characteristics of the three types, but no significant difference of the 5-year survival rates was found among the three types of patients with curative resection. CONCLUSION: On the data, the distribution of the three types of tumor was found to be different from that reported in Western countries and in Japan; and the three types of patients who had undergone curative resection were found to have similar 5-year survival rates.


Subject(s)
Adenocarcinoma/classification , Esophageal Neoplasms/classification , Esophagectomy , Esophagogastric Junction , Lymph Node Excision , Stomach Neoplasms/classification , Adenocarcinoma/mortality , Adenocarcinoma/surgery , Barrett Esophagus/pathology , Cardia , China/epidemiology , Esophageal Neoplasms/mortality , Esophageal Neoplasms/pathology , Esophageal Neoplasms/surgery , Esophagogastric Junction/surgery , Female , Gastrectomy , Gastric Mucosa/pathology , Humans , Lymph Nodes/pathology , Male , Metaplasia , Middle Aged , Morbidity , Postoperative Complications/etiology , Stomach Neoplasms/mortality , Stomach Neoplasms/surgery , Survival Rate
16.
Protein Pept Lett ; 13(4): 405-10, 2006.
Article in English | MEDLINE | ID: mdl-16712519

ABSTRACT

L-Arginine kinase (AK; ATP:L-arginine N-phosphotransferase; EC 2.7.3.3) catalyzes the reversible transphosphorylation between N-phospho-L-arginine (PArg) and ATP thus buffering cellular ATP levels. AK was purified from the leg muscle of the locust Migratoria manilensis by Sephacryl S-200 HR gel filtration chromatography and DEAE Sepharose CL-6B fast flow anion exchange chromatography to an apparent homogeneity with a recovery of 80%. The enzyme behaved as monomeric protein with molecular mass of about 40 kD, and had a pH and temperature optimum of 8.6 and 30 degrees C, respectively, and a pI of about 6.3. The Michaelis constants for synthesis of PArg are 0.936 and 1.290 mM for L-arginine and ATP, respectively and k(cat)/K(m)(Arg) 174. The activity of AK required divalent cations such as Mg(2+) and Mn(2+). In the presence of Cu(2+) and Zn(2+), AK activity was greatly inhibited. The intrinsic protein fluorescence emission maximum at 330 nm using the excitation wavelength at 295 nm suggested that tryptophan residues are below the surface of the protein and not exposed to solvent.


Subject(s)
Arginine Kinase/isolation & purification , Amino Acids/analysis , Animals , Arginine Kinase/chemistry , Arginine Kinase/metabolism , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Grasshoppers/enzymology , Hydrogen-Ion Concentration , Kinetics , Muscle, Skeletal/enzymology , Spectrometry, Fluorescence , Temperature
17.
Article in Chinese | MEDLINE | ID: mdl-16562664

ABSTRACT

OBJECTIVE: To construct eukaryotic expression plasmid of porcine CCK gene pIRES2-EGFP/CCK and express it in COS-7 cells and hamsters. Methods The aimed segments were obtained from intermediate vector pMD18-T/CCK and were inserted into an eukaryotic expression plasmid pIRES2-EGFP to construct a recombinant expression plasmid pIRES2-EGFP/CCK. The recombinant expression plasmid was transfected into COS-7 cells by liposome-mediated gene transfer method and was observed through fluorescence microscope. The plasmid was injected into the skeletal muscle of hamsters directly to detect the expression of the recombinant plasmid in vivo. RESULTS: A recombinant eukaryotic expression plasmid pIRES2-EGFP/CCK was successfully constructed. Green fluorescent protein could be detected in the transfected COS-7 cells 24, 48, and 72 hours after the transfection. On the 4th day postinjection into the skeletal muscle of hamsters, the protein could be detected at the injection site and the fluorescence intensity became much stronger on the 14th day than that on the 4th day. On the 42nd day the protein level increased. The green fluorescence protein was never expressed in the untransfected cells. CONCLUSION: The porcine CCK gene eukaryotic expression plasmid pIRES2-EGFP/CCK is constructed successfully, and is expressed in mammal COS-7 cells and hamsters in vivo. The research paves the way for the cross immunity therapy of hamster pancreatic carcinoma.


Subject(s)
Cholecystokinin/biosynthesis , Green Fluorescent Proteins/biosynthesis , Recombinant Fusion Proteins/biosynthesis , Animals , Base Sequence , COS Cells , Cancer Vaccines/therapeutic use , Chlorocebus aethiops , Cholecystokinin/genetics , Cricetinae , Eukaryotic Cells/metabolism , Green Fluorescent Proteins/genetics , Molecular Sequence Data , Muscle, Skeletal/metabolism , Pancreatic Neoplasms/therapy , Plasmids , Recombinant Fusion Proteins/genetics , Swine , Transfection
18.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 36(5): 609-12, 2005 Sep.
Article in Chinese | MEDLINE | ID: mdl-16235518

ABSTRACT

OBJECTIVE: To construct eukaryotic expression plasmid of porcine CCK gene pIRES2-EGFP/ CCK and express it in COS-7 cells and hamsters. METHODS: The aimed segments were obtained from intermediate vector pMD18-T/CCK by the method of restricted enzymatic resection and were inserted into a eukaryotic expression plasmid pIRES2-EGFP to construct a recombinant expression plasmid pIRES2-EGFP/CCK. The recombinant expression plasmid was transfected into COS-7 cells by liposome-mediated gene transfer method and observed through Fluorescence microscopy. The plasmid was injected into the skeletal muscle of hamsters directly to detect the expression of the recombinant plasmid in vivo. RESULTS: A recombinant eukaryotic expression plasmid pIRES2-EGFP/CCK was successfully constructed. Green fluorescent protein could be detected in the transfected COS-7 cells 24, 48, and 72 hours post transfection and the expression of green fluorescent protein reached its peak 72 h post transfection. The green fluorescent protein could be detected at the injection site on the 4th day post injection and the fluorescence intensity became stronger on the 14th day. The level of fluorescence became ever stronger on the 42nd day. No expression of green fluorescence was detected in the control group. CONCLUSION: Porcine CCK cDNA eukaryotic expression plasmid pIRES2-EGFP/CCK has been successfully constructed and expressed in mammal cells COS-7 and hamster in vivo. The research paved the way for cross immunity therapy of hamster pancreatic carcinoma.


Subject(s)
Cholecystokinin/genetics , Green Fluorescent Proteins/genetics , Recombinant Fusion Proteins/biosynthesis , Animals , COS Cells , Chlorocebus aethiops , Cholecystokinin/biosynthesis , Cricetinae , Female , Green Fluorescent Proteins/biosynthesis , Humans , Immunotherapy , Mesocricetus , Pancreatic Neoplasms/therapy , Plasmids/genetics , Recombinant Fusion Proteins/genetics , Swine , Transfection
19.
Shi Yan Sheng Wu Xue Bao ; 37(6): 494-500, 2004 Dec.
Article in Chinese | MEDLINE | ID: mdl-15789770

ABSTRACT

A cDNA fragment that encodes auxin-binding protein 1 was amplified by, reverse polymerase chain reaction from ovary of cucumber. Its expression signals were weak in the ovary of 1 d before anthesis, while got strong in 2, 4 and 6 d after pollination. Among the unpollinated ovary of 2 d after anthesis, those that got enlarged had strong expression signals; the others that were wilting had weak signals. This indicated that auxin-binding protein 1 gene possibly play a role in cucumber fruit development. When auxin-binding protein 1 gene of Arabidopsis was transformed into cucumber, the parthenocarpic rate of transgenic plants was 31.7%, higher than the control. This result showed that the sensibility to auxin was increased in transgenic plants.


Subject(s)
Cucumis sativus/growth & development , Fruit/growth & development , Plant Proteins/physiology , Plants, Genetically Modified/growth & development , Receptors, Cell Surface/physiology , Amino Acid Sequence , Arabidopsis/genetics , Base Sequence , Cucumis sativus/drug effects , Cucumis sativus/genetics , Fruit/drug effects , Fruit/genetics , Gene Expression Regulation, Developmental/drug effects , Gene Expression Regulation, Developmental/genetics , Gene Expression Regulation, Plant/drug effects , Gene Expression Regulation, Plant/genetics , Indoleacetic Acids/pharmacology , Molecular Sequence Data , Plant Proteins/chemistry , Plant Proteins/genetics , Plants, Genetically Modified/drug effects , Plants, Genetically Modified/genetics , Receptors, Cell Surface/chemistry , Receptors, Cell Surface/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment
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