2.Cloning, Sequence Analysis and Expression of dhaT Gene from Citrobacter freundii and Purification and Property of Corresponding Recombinant Enzyme
Xiang-Hui QI ; Zhao-Fei LUO ; Yu-Tuo WEI ; Fa-Zhong CHEN ; Shan-Shan WANG ; Shou-Hai HOU ; Dong-Qing LIAO ; Ri-Bo HUANG ;
China Biotechnology 2006;0(07):-
1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.
3.Surgical repair of ruptured sinus of Valsalva aneurysm to right atrium.
Hong-Wei GUO ; Qian CHANG ; Cun-Tao YU ; Xiao-Gang SUN ; Xiang-Yang QIAN ; Yong-Bo WU ; Jun FENG ; Sheng-Shou HU
Chinese Journal of Surgery 2010;48(15):1158-1160
OBJECTIVESTo summarize the experience of surgical repair of ruptured sinus of Valsalva aneurysm to right atrium and to compare the difference between through right atrium repair and transaortic combined with right atrium approach.
METHODSBetween January 2004 and December 2009, 53 patients with ruptured sinus of Valsalva aneurysm to right atrium underwent surgical repair. There were 35 male and 18 female, aged from 15 to 63 with a mean of (33 ± 9) years. Repair through right atrium had undergone in 40 patients (group I), while transaortic combined with right atrium approach in 13 patients (group II). Surgical results between the two group and group were compared in cardiopulmonary bypass time, clamp aorta time, mechanical ventilation time, ICU time and postoperative stay time.
RESULTSThere were no significant differences between two groups in cardiopulmonary bypass time [(86 ± 29) min vs. (96 ± 30) min], clamp aorta time [(59 ± 29) min vs. (71 ± 25) min], mechanical ventilation time [(9 ± 4) h vs. (16 ± 23) h], ICU time [(35 ± 23) h vs. (35 ± 23) h], postoperative stay time [(7.1 ± 0.9) d vs. (7.7 ± 2.8) d] (P > 0.05). Follow-up was performed from 1 to 64 months, with a mean of (32 ± 21) months. There was no death during follow up. One needed operation due to severe aortic valve regurgitation. One combined with coronary artery disease used medication. Heart function (NYHF) of the other patients were I and II degree during follow up.
CONCLUSIONSSurgical repair of ruptured sinus of Valsalva aneurysm to right atrium shows good result. There is no significant difference between through right atrium repair and transaortic combined with right atrium approach.
Adolescent ; Adult ; Aorta ; surgery ; Aortic Rupture ; surgery ; Female ; Follow-Up Studies ; Heart Atria ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sinus of Valsalva ; Treatment Outcome ; Young Adult
4.Expression and SNP analysis of BRD7 gene in acute leukemia cells.
Xue-yuan TANG ; Fen LIU ; Ya PENG ; Bo XIANG ; Cheng-hong WANG ; Chen-jiao YAO ; Shou-rong SHEN
Journal of Central South University(Medical Sciences) 2008;33(8):645-650
OBJECTIVE:
To evaluate the expression level of BRD7 gene in bone marrow mononuclear cells (BMNCs) in patients with acute leukemia (AL) and to analyze BRD7 single nucleotide polymorphism(SNP).
METHODS:
RT-PCR was used to detect BRD7 expression in patients with AL and normal bone marrow subjects. Single-strand conformation polymorphism and DNA sequence analysis were also used to identify BRD7 mutation or SNP to investigate the relation between BRD7 and AL.
RESULTS:
BRD7 mRNA in BMNCs from 52 patients with AL and 30 control subjects was expressed. The mRNA relative expression of BRD7 in patients with AL was higher than that of the control group (P=0.001). Three SNPs (C657A,C495T and A737G) in BRD7 gene coding region (447 approximately 844 bp) were found, and A737G was coupled with C495T . The allele frequencies of SNP C657A were not significantly different between AL and the control group. The genotype and the allele frequencies of the 2 coupled SNPs were significantly different (P<0.01). But there was no significant discrepancy among the mRNA expression levels of AA, AG, and GG genotypes in the leukemia group (P>0.05).
CONCLUSION
Expression of BRD7 gene is up-regulated in AL cells. The 2 coupled SNPs (C495T and A737G ) in BRD7 gene coding region (447 approximately 844 bp) are correlated with AL, indicating that SNPs may be one of the genetic susceptibility factors of AL.
Adolescent
;
Adult
;
Aged
;
Amino Acid Sequence
;
Base Sequence
;
Child
;
Chromosomal Proteins, Non-Histone
;
biosynthesis
;
genetics
;
Female
;
Gene Frequency
;
Humans
;
Leukemia, Myeloid, Acute
;
genetics
;
Male
;
Middle Aged
;
Molecular Sequence Data
;
Polymorphism, Single Nucleotide
;
Precursor Cell Lymphoblastic Leukemia-Lymphoma
;
genetics
;
RNA, Messenger
;
biosynthesis
;
genetics
6.Postoperative diaphragmatic paralysis after cardiac surgery in children: incidence, diagnosis and surgical management.
Yan-Bo ZHANG ; Xu WANG ; Shou-Jun LI ; Ke-Ming YANG ; Xiang-Dong SHENG ; Jun YAN
Chinese Medical Journal 2013;126(21):4083-4087
BACKGROUNDCardiac surgery for congenital heart disease covers a wide spectrum from simple to complex cardiac and extracardiac malformations. Innovations in pediatric cardiac surgery and perioperative care over the past decades have allowed surgical correction or at least palliation in almost all complex congenital heart defects in the first years of life. Diaphragmatic paralysis (DP) due to phrenic nerve injury after congenital cardiac surgery is an important respiratory complication resulting with respiratory insufficiency, lung infections, prolonged hospital stay time and even death.
METHODSBetween April 2001 and December 2010, among patients undergoing cardiac surgery for congenital heart disease, postoperative DP was diagnosed in 47/10 200 (0.46%) patients. Diaphragmatic placation was performed in 37/47 patients. DP was suspected in children who failed to wean from mechanical ventilation or in those with persistent respiratory distress when there is no cardiac cause. Decreased respiratory sounds in auscultation, paradoxical breathing during spontaneous ventilation and elevated hemidiaphragm on chest X-ray led us to use fluoroscopy, ultrasound and/or electromyogram (EMG). When chest X-rays did not have a diagnostic value in patients with persistent respiratory distress, bilateral DP was suspected and immediate fluoroscopy of EMG was performed for diagnosis. In all patients, diaphragmatic placation was performed using a thoracic approach, through the sixth or seventh intercostals space with lateral thoracotomy.
RESULTSA total of 47 patients (21 females and 26 males) with a median age of 7.21 months (range 0.27-71 months) were diagnosed DP after cardiac surgery. The incidence of DP was 0.46% after cardiac surgery. The paralysed hemidiaphragm was left side in 26/47 (55.3%), right side in 17/47 (36.2%) and bilateral in 4/47 (8.5%) cases. The assisted ventilation time after cardiac surgery was (450±216) (116-856) hours. The median time from cardiac surgery to surgical placation was (24±14) (5-56) days. No patient died in this study. The follow-up period was (26.2±16.8) months. The position of the plicated diaphragm was normal on chest X-ray, in all plicated survivors within the 1st, 6th and 12th months after discharge.
CONCLUSIONSDP caused by phrenic nerve injury during surgical intervention for congenital heart disease is an important risk factor in terms of morbidity during the postoperative period. Diaphragmatic placation appears a good option, especially in newborns and small children, to wean patients from mechanical ventilation and to prevent long-term side effects of mechanical ventilation.
Cardiac Surgical Procedures ; adverse effects ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Postoperative Complications ; Respiratory Paralysis ; epidemiology ; etiology ; surgery
7.Endothelial nitric oxide synthase gene polymorphisms associated with susceptibility to high altitude pulmonary edema in Chinese railway construction workers at Qinghai-Tibet over 4 500 meters above sea level.
Sun YU-JING ; Fang MING-WU ; Niu WEN-QUAN ; Li GUANG-PING ; Liu JING-LIANG ; Ding SHOU-QUAN ; Xu YING ; Yu GUO-SHU ; Dong JIAN-QUN ; Pan YUN-JUN ; Dong WEI-YA ; Wang TIAN ; Cao JING-WEN ; Li XIAO-BO ; Wang ZHONG-XIANG ; Yu GUANG-XUE ; Sun HUI-CHENG ; Jia ZHONG-HOU ; Liu JUN ; Wang XIAO-MING ; Si QIN ; Wu QI-XIA ; Zhou WEN-YU ; Zhu TONG-CHUN ; Qiu CHANG-CHUN
Chinese Medical Sciences Journal 2010;25(4):215-221
OBJECTIVETo examine whether the polymorphisms of endothelial nitric oxide synthase (eNOS) gene are associated with the susceptibility to high altitude pulmonary edema (HAPE) in Chinese railway construction workers at Qinghai-Tibet where the altitude is over 4 500 m above sea level.
METHODSA case-control study was conducted including 149 HAPE patients in the construction workers and 160 healthy controls randomly recruited from their co-workers, matching the patients in ethnicity, age, sex, lifestyle, and working conditions. Three polymorphisms of eNOS gene, T-786C in promoter, 894G/T in exon 7, and 27bp variable number tandem repeat (VNTR) in intron 4, were genotyped using polymerase chain reaction (PCR) and confirmed with DNA sequencing.
RESULTSThe frequencies of 894T allele and heterozygous G/T of the 894G/T variant were significantly higher in HAPE patients group than in the control group (P=0.0028 and P=0.0047, respectively). However, the frequencies of the T-786C in promoter and the 27bp VNTR in intron 4 were not significantly different between the two groups. Haplotypic analysis revealed that the frequencies of two haplotypes (H3,T-T-b, b indicates 5 repeats of 27 bp VNTR; H6, C-G-a, a indicates 4 repeats of 27 bp VNTR) were significantly higher in HAPE patients (both Pü0.0001). On the contrary, the frequencies of H1 (T-G-b) and H2 (T-G-a) were lower in HAPE patients than in healthy controls (both Pü0.001).
CONCLUSIONSTwo haplotypes (T-T-b and C-G-a) may be strongly associated with susceptibility to HAPE. Compared with the individual alleles of eNOS gene, the interaction of multiple genetic markers within a haplotype may be a major determinant for the susceptibility to HAPE.
Adolescent ; Adult ; Altitude ; Base Sequence ; Case-Control Studies ; DNA Primers ; Genotype ; Haplotypes ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; genetics ; Occupational Diseases ; enzymology ; genetics ; Polymorphism, Genetic ; Pulmonary Edema ; enzymology ; genetics ; Tibet ; Young Adult
8.High Physical Activity is Associated with an Improved Lipid Profile and Resting Heart Rate among Healthy Middle-aged Chinese People.
Bo HU ; Xiao Yu LIU ; Yao ZHENG ; Hong Min FAN ; Su Feng YIN ; Chun Yue GUO ; Yun LI ; Shou Ling WU ; Fu Min FENG ; Ju Xiang YUAN
Biomedical and Environmental Sciences 2015;28(4):263-271
OBJECTIVETo investigate the effects of physical activity (PA) on dyslipidemia and elevated resting heart rate (RHR) in a large-scale cross-sectional study in China.
METHODSWe recruited community-based individuals who were 40-60 years old using a cluster sampling method. The PA levels of the participants were classified as low, moderate, or high, using the International Physical Activity Questionnaire. Dyslipidemia was defined as the detection of abnormalities in lipid indicators, and 4 lipid parameters were evaluated using fasting blood samples. Multivariate logistic regression analyses were used to evaluate the associations of PA with dyslipidemia and RHR.
RESULTSA total of 10,321 participants (38.88% men) were included in this study. The percentages of individuals with high, moderate, and low PA levels were 46.5%, 43.9%, and 9.6%, respectively. In both men and women, high PA provided odds ratios of 0.88 [95% confidence interval (CI): 0.83, 0.94] for dyslipidemia and 0.82 (95% CI: 0.73, 0.92) for elevated RHR, compared to participants with low PA.
CONCLUSIONOur data suggested that substantial health benefits (related to dyslipidemia and elevated RHR) occurred at higher intensity PA, with greater energy consumption, in middle-aged Chinese people, and particularly in men.
Adult ; China ; epidemiology ; Cross-Sectional Studies ; Dyslipidemias ; epidemiology ; etiology ; Female ; Heart Rate ; Humans ; Lipids ; blood ; Logistic Models ; Male ; Middle Aged ; Motor Activity ; Odds Ratio ; Risk Factors ; Sex Factors
10.Comparison on the efficacy of Chinese-made novel-designed mechanical-locked and elastic self-locked transcatheter edge-to-edge repair system in the treatment of patients with functional mitral regurgitation.
Da ZHU ; Shou Zheng WANG ; Zhi Ling LUO ; Jia Hua PAN ; Ke YANG ; Chun Mei XIE ; Yong Yan TANG ; Hong Bo YANG ; Mi Er MA ; Jian Bin GAO ; Xiang Bin PAN
Chinese Journal of Cardiology 2023;51(8):832-837
Objective: To evaluate and compare the short-term efficacy of domestic mechanical-locked (Clip2Edge) and elastic self-locked (ValveClip) transcranial mitral valve edge-to-edge interventional repair (TEER) devices in the treatment of functional mitral regurgitant valves. Methods: In this retrospective non-randomized comparative study, patients underwent TEER procedure in Fuwai Yunnan Cardiovascular Disease Hospital from May 2022 to April 2023 for heart failure combined with moderate to severe or severe functional mitral valve were divided into Clip2Edge and ValveClip groups based on the TEER system used. Baseline, perioperative, and postoperative 30 d follow-up data were collected and compared between the two groups. The primary outcome was the success rate on the 30 d post operation, while secondary outcomes included immediate postoperative technical success rate and the incidence of all-cause mortality on the 30 d post operation, readmission rate of acute heart failure, cerebral infarction, severe bleeding, and other serious adverse events rates. Results: A total of 60 patients were enrolled, 34 patients were in the Clip2Edge group and 26 in the ValveClip group, mean age was (63.8±9.3) years, and 24 patients (40%) were female. There were no significant differences in baseline data of age, cardiac function, comorbidities, mitral regurgitation 4+(19(73%) vs. 29(85%)), the end-diastolic volume of left ventricle ((220.8±91.2) ml vs. (210.8±71.7) ml) between the two groups (all P>0.05). The technical success rate immediately after the procedure was 100%. There were no readmission of acute heart failure, death, cerebral infarction, severe bleeding, and other serious adverse events up to the 30 d follow-up. Device success rate was similar between the ValveClip group (24 cases (100%)) and the Clip2Edge group (27 cases (96%)) (P>0.05). Conclusion: Both types of novel domestic TEER devices are safe and feasible in treating patients with functional mitral regurgitation.
Humans
;
Female
;
Middle Aged
;
Aged
;
Male
;
Mitral Valve Insufficiency/etiology*
;
Retrospective Studies
;
East Asian People
;
Heart Valve Prosthesis Implantation
;
Treatment Outcome
;
China
;
Heart Failure/etiology*
;
Cardiac Catheterization