Six 89bp primers were designed on the base of the cDNA sequence encoding the human leptin reported on the NCBI. The synthetic gene with 464bp encoding rhLep was obtained by SOE ( splicing by overlap extension) PCR. The expression vector pET22b(+)/rhLep was constructed and transformed into E. coli BL21 (DE3). The rhLep protein was expressed as inclusion bodies with the yield of more than 50% of total bacterial proteins after IPTG induction. The rhLep protein, which has a molecular weight about 16kD, was purified by Ni2+ affinity chromatography column and identified by SDS-PAGE. The MTT Assay shows that rhLep promotes EC304 cells growth at the low concentration of 10ng/mL to 30 ng/mL, and rhLep appears cytotoxic to EC304 cells with the high dose of 50ng/mL to 225ng/mL. The viability of EC304 cells decreases to 1.2% with the concentration of 225ng/mL of rhLep. The massive apoptosis of rhLep on EC304 cells is observed by AO-staining under fluorescent microscope. All these results would lay the foundation for the further study of its biological functions in vitro and in vivo.
Apoptosis ; drug effects ; Escherichia coli ; genetics ; Humans ; Leptin ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology

OBJECTIVETo evaluate the effect of etodimate infusion on serum cortisol in patients undergoing radical resection of lung cancer operations during the perioperative period.

METHODSForty ASA I-II patients undergoing radical resection of lung cancer were randomly divided into etomidate group (Group E) and propofol group (Group P) (n=20). The serum cortisol was measured at 8:00 am (T(0)) before anesthesia, 4:00 pm (T(1)) on the day of operation and 24 h after the operation (T(2)) by radioimmunoassay.

RESULTSCompared with that at T0, the serum level of cortisol significantly increased at 24 h after the operation in both groups (P<0.01); serum cortisol decreased lightly at T1, which was not statistically significant (P>0.05), and remained higher than the normal level. At each of the time points, serum cortisol levels were comparable between the two groups (P>0.05).

CONCLUSIONEtomidate infusion can not inhibit the synthesis of cortisol in patients undergoing radical resection of lung cancer.

Adult ; Anesthesia, General ; Etomidate ; administration & dosage ; Female ; Humans ; Hydrocortisone ; blood ; Lung Neoplasms ; blood ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods ; Postoperative Period ; Young Adult

UNLABELLEDTo prepare a novel fusion protein (tTF/SA) as a universal effector for targeting therapy of blood coagulation and to analyze its biological activities. The fusion gene tTF/SA was constructed by PCR, then inserted into expression vector pET22 b (+), and expressed in E. coli BL21 (DE3). The fusion protein was purified using Nickel-affinity chromatography column. The activities of tTF moiety of the fusion protein were analyzed by clotting assay and FX activation assay, and the binding activities of Streptavidin(SA) to Biotin(B) were analyzed using ELISA.

RESULTSThe recombinant plasmid tTF/SA/pET22 b (+) with the correct sequence was obtained. The fusion gene tTF/SA was expressed with high yield in E. coli BL21 (DE3). The purified fusion protein retain the abilities of activating FX, inducing blood coagulation, and binding Biotin. The fusion gene tTF/SA was successfully expressed in E. coli BL21 (DE3). The recombinant tTF/SA proteins retain the activities of TF and SA. The multitarget therapy of selectively inducing thrombosis in tumor blood vessels can be achieved by the combination of tTF/SA, as a universal effector, and biotinlated carriers directing to tumor blood vessels.

Animals ; Binding, Competitive ; drug effects ; Biotin ; metabolism ; Blood Coagulation ; drug effects ; physiology ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; Mice ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Streptavidin ; genetics ; metabolism ; pharmacology ; Thromboplastin ; genetics ; metabolism ; pharmacology ; Time Factors

OBJECTIVETo study the progression of myopia in school-age children over the past 12 years and factors influencing myopia progression.

METHODSA total of 4569 cases of 5 to 12-year-old children who had refractive examinations in the Third Xiangya Hospital, Central South University between January 2000 and December 2011 were enrolled in this study. The children had no family history of congenital high myopia or other eye diseases. Myopia progression was evaluated when the children were re-examined. The refractive state of each child was measured with cyclopiegic retinoscopy.

RESULTSThe mean spherical equivalent (SE) myopia was-2.0±1.7 D between January 2000 and December 2011. There was no statistical difference in yearly myopia progression between different years. The average age of the myopic children decreased from 10.1 in 2000 to 8.9 years old in 2011 (P<0.05). Mean myopia progression was -0.6±0.7 D per year from 2000 to 2011. Myopia progression reduced gradually in 5 to 8-year-olds (P<0.05), however, it accelerated between ages 9 and 11 years. Myopia progression in 10- and 11-year-olds was significantly greater than in 7- and 8-year-olds (P<0.01). The multiple linear regression analysis demonstrated that age and baseline myopic refraction were positively related to myopia progression.

CONCLUSIONSThere was no obvious change in the yearly myopia progression of the children over the past 12 years. The mean age of myopia occurrence became younger with time. More preventive measures are needed to ward off high myopia in children with moderate myopia, especially those aged over 10 years.

Adolescent ; Child ; Child, Preschool ; Disease Progression ; Female ; Humans ; Linear Models ; Male ; Myopia ; etiology ; Retrospective Studies

Objectives: To investigate whether remote ischemic conditioning (RIC) applied to patients with ST-segment elevation myocardial infarction (STEMI) before percutaneous coronary intervention (PCI) could affect circulating miR-208b level or not. Methods:Patients diagnosed with STEMI undergoing PCI from January 2016 to July 2017 were enrolled from the Department of Cardiology, Affiliated Zhongshan Hospital of Dalian University.The patients were randomly allocated to two groups: (1) control group (n=25), PCI alone; (2) RIC group (n=50), PCI combined with RIC (three cycles of 5 min inflation and 5 min deflation of the right lower limb with blood pressure cuff performed before reperfusion). Serum miR-208b was measured before and immediately, at 24 h, and 48 h after PCI with real-time quantitative polymerase chain reaction. Results: The expression of miR-208b was significantly higher immediately post PCI than that before operation in the control group (84.1±9.0 vs 77.8±9.4; P=0.032), while it was significantly lower immediately post PCI than that before operationin RIC group (71.0±9.3 vs 77.4±8.8; P=0.028).miR-208b level was similar before PCI between the control and RIC groups (P=0.874), which was significantly reduced immediately post PCI in RIC group as compared with the control group (P=0.021).The peak value of creatine kinase isoenzyme (CK-MB) in the limb RIC group was significantly lower than that in the control group ([135.2±18.6] U/L vs [167.7±17.2] U/L; P=0.038).The area under the CK-MB curve of the RIC group was significantly smaller than that of the control group ([3 060.7±17.1] U/L vs [3 635.9±15.1] U/L); P=0.047]. The left ventricular ejection fraction (LVEF) in RIC group was significantly higher than that in the control group ([57.8±7.8]% vs [51.9±7.9]%; P=0.003) post PCI. The expression level of serum miR-208b was positively correlated with CK-MB AUC in RIC group (r=0.498, P<0.001). Conclusions: RIC of the lower limb prior to PCI could reduce miR-208b level and improve cardiac functionin STEMI patients.

OBJECTIVETo provide materials for the study of the function of ESC42 protein specifically expressed in the human epididymis.

METHODSThe ESC42 gene was amplified from the human epididymis cDNA library by PCR and then cloned into prokaryotic expression vector pGEX-4T-1, expressed and purified by recombinant DNA techniques. The specificity of ESC42 protein was identified by Western blot and MALDI-TOF-MS. The database was searched by Ms-Fit.

RESULTSThe recombinant plasmid expressed a Mr 38 x 10(3) fusion protein in E. coli at a level of 30% of the total protein, and the purity was as high as 99%. The ESC42 protein was identified by ESC42 monoclonal antibody and its molecular weight was 11 978.12, tested by MALDI-TOF-MS. The peptide mass fingerprint analysis showed that the coverage rate of the sequence reached 48% with 100% matching. The motif scan in Prosite database reveal that ESC42 belonged to the beta-defensin family and had antibacterial activity.

CONCLUSIONObtaining high purity of rhESC42 protein may lay a foundation for the study of its functions.

Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; immunology ; Cloning, Molecular ; Defensins ; biosynthesis ; genetics ; immunology ; Epididymis ; metabolism ; Escherichia coli ; genetics ; Gene Library ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; immunology

OBJECTIVEWU polyomavirus (WUPyV), a new member of the genus Polyomavirus in the family Polyomaviridae, has been found to be associated with respiratory tract infections recently. But the role of the WUPyV as agents of human disease remains uncertain. We sought to describe the detection and clinical characterization of WUPyV in acute respiratory tract infection in children.

METHODFrom July 2008 through June 2009, nasopharyngeal aspirates were collected from 771 children who were hospitalized with acute respiratory tract infection in Second Affiliated Hospital of Shantou University Medical College, and from 82 asymptomatic children who visited the health checkup clinic. WUPyV was detected by using PCR technology and was identified by using DNA sequencing. All WUPyV-positive specimens were screened for 9 common viruses [influenza A and B, respiratory syncytial virus (RSV), parainfluenza virus (PIV) 1 and 3, human metapneumovirus, human bocavirus, adenovirus and rhinovirus] by using PCR or RT-PCR. The clinical data of WUPyV infection were collected and analyzed.

RESULTIn this study, fifteen of the 771 tested specimens with acute respiratory tract infection were positive for WUPyV, the positive rate was 1.95% and all of the asymptomatic children who visited the health checkup clinic were negative. Of the 15 cases who were positive for the virus, the age range was 2 to 48 (mean 18.8) months, 9 (60%) were male and 6 (40%) were female. WUPyV was the sole virus detected in 9 specimens (60%) from patients with acute respiratory tract infection. WUPyV was associated with the co-infection with another respiratory virus in 6 of 15 (40%) cases, most frequently with RSV (n = 4), followed by adenovirus (n = 1) and rhinovirus (n = 1). The most common clinical findings in the patients with WUPyV were cough, fever and wheezing. The most frequent diagnoses were pneumonia (n = 8), bronchiolitis (n = 4), upper respiratory tract infections (n = 2) and bronchitis (n = 1). A severe case was complicated with viral encephalitis.

CONCLUSIONWUPyV may be a respiratory pathogen because it was the sole virus detected in 9 specimens from patients with respiratory illness and all of the asymptomatic controls were negative. The most common clinical findings are cough and wheezing. Young children may be susceptible to infection with this virus and occasionally the infection with this virus may cause severe disease. More comprehensive and in-depth studies are required to prove the pathogenicity of these viruses.

Child ; Child, Preschool ; Female ; Genes, Viral ; Humans ; Infant ; Infant, Newborn ; Male ; Polymerase Chain Reaction ; Polyomavirus ; genetics ; isolation & purification ; Polyomavirus Infections ; physiopathology ; virology ; Respiratory Tract Infections ; virology

Objective Vasoconstriction and vascular hypersensitivity to serotonin were previously shown in animal models of adventitia injury. We investigated the contribution of angiotensin Ⅱ(Ang Ⅱ)/Ang Ⅱ receptors and oxidative stress to vascular contractility and reactivity in this model. Methods Wistar Kyoto rats were divided into 3 groups: normal(n =6, no any intervention, only for measuring the serum Ang d-1). After one week of treatment, adventitia injury was induced by positioning a silicone collar around the right carotid artery for one week. Blood flow and vascular reactivity to serotonin were determined one week after injury, the blood from left ventricle was taken to measure the serum Ang Ⅱ concentration by ELISA,and carotids were harvested for morphometry and Western blot analysis. Results Adventitia injury induced lumen cross-sectional area reduction(- 44% vs. - 5%), media diameter increase(62% vs. 10%),blood flow reduction[(2. 79 ± 0. 22)vs.(4. 33 ± 0. 84)ml/min]were significantly attenuated by valsartar. The increased vascular reactivity sensitivity to serotonin in vehicle group was also significantly reduced in valsartan group. Serum Ang Ⅱ concentration was significantly increased in vehicle group [(45.21 ± 4. 52)pg/ml vs.(19. 83 ± 0. 5)pg/ml in normal rats, P = 0. 0148]and the expression of Ang Ⅱtype 1(AT1)receptor, Ang Ⅱ type 2(AT2)receptor, as well as p22pbox in collared arteries were significantly upregulated. Valsartan did not affect the AT1 receptor expression but further increased serum Ang Ⅱ concentration[(89. 73 ±20. 44)pg/ml vs.(45.21 ±4. 52)pg/ml, P =0. 001], and AT2 receptor expression, while downregulated p22phox expressions. Conclusions Collar-induced adventitia injury resulted in chronic vsoconstriction and vascular hypersensitivity to serotonin via increased serum Ang Ⅱ level,upregulated Ang Ⅱ receptors expression in the vascular well, and activated local oxidative stress. These changes could be blocked by valsartan suggesting a crucial role of Ang Ⅱ/Ang Ⅱ receptors on vascular contractility and reactivity changes in this model.

Objective To provide data for the control and prevention of hepatitis B and HBV surface antigen(HBsAg)status among the appliances and practitioners working in the public service places.Methods 63 beauty parlors,barber shops and bathing centers selected under stratified randomization sampling method and 682 workers were investigated through questionnaire.HBsAg from the appliances of the public service places and employee was detected by RIA.Results Two main sanitizing modes that including alcohol cleaning(34.60%)and ultraviolet light disinfection(30.79%)were used.The rates of testing on HBsAg among the appliances were 2.13% at the public service places,and were 0.63%,2.67% and 3.70% in large-.medium-and small-sized appliances respectively.The rate of testing on HBsAg on large-,medium-and small-sized appliances were statistically different(χ2=6.68,P＜0.05).The positive rates of HBsAg on the appliances of beauty parlors,barbering shops and footbath inns were 2.97%,0.61% and 3.42% respectively.People working in different service sites had different rates of HBsAg:those who worked at the‘acne needle'and the forceps were 5.13% and 4.17%.The positive rate of HBsAg among the workers in the public service places was 7.13%.The rates of HBsAg among the workers in large-,medium-and small-sized public service places were 7.34%,8.33% and 2.94% respectively.The rates of HBsAg among the workers in beauty parlors,barbering shops,footbath inns and bathing centers were 9.01%,6.37%,4.35% and 7.29% respectively.HBsAg positive rates were different among the workers working at different service sites:13.33% at tattoo business.12.68% in pedicures workers and 8.03% in massagists.Conclusion It is important to improve the sanitizing management of the appliances used in the public service places and to improve the knowledge,attitude,as well as practice of vaccination on hepatitis B among those populations.

OBJECTIVETo describe the epidemiology of neural tube defects (NTDs) in high- and low-prevalence areas of China.

METHODSBirth defects surveillance data, collected from 1992 through 1994 was analyzed. These data were collected as part of the Sino-American cooperative project on NTDs prevention. We classified NTDs as anencephaly, encephalocele, high-level and low-level spina bifida (SB) according to location of the lesion (high vs low) and whether the defect was isolated or occurred in association with other birth defects. Rates were compared in the high-prevalence (North) region and the low-prevalence (South) region, after adjusted for classification, urban and rural, season and sex, and calculated the adjusted rate of NTDs.

RESULTSAmong seven hundred and eighty-four NTDs cases in 326 874 recorded births (include in livebirth, stillbirth and fetal death with a gestational age of at least 20 weeks), the overall NTDs prevalence in the North was 5.57/1,000 births, and in the South was 0.88/1 000. There were also significant differences in the prevalence of anencephaly, encephalocele, high-level and low-level SB between North (0.97, 0.49, 2.75 and 1.11/1,000 birth) and South (0.36, 0.15, 0.21 and 0.14/1,000 birth) (P < 0.01), with adjusted prevalences in the North 3 - 7 times higher than those in the South. There were significant difference between urban (2.04) and rural areas (6.57/1,000 birth) in the North (P < 0.01), urban (0.52) and rural areas (0.95/1,000 birth) in the South (P < 0.05). Adjusted prevalence rates in the rural were 3 - 4 times higher than those of urban in the North and 1.6 - 1.9 times higher than in the South; The seasonal rate of high-level SB increased between September and November in the North (3.44/1,000 birth), while the seasonal rate of anencephaly decreased between September and November (0.18/1,000 birth) in the South. However there were no seasonal changes in other classified NTDs both in the South and North.

CONCLUSIONSThe birth prevalence of NTDs in the North of China was the highest in the world. There were significant differences between the North and the South, urban and rural. There was seasonal change in high-level SB in the North, which was in accordance to the phenotype of NTDs. It was suggested that there might exist etiological heterogeneity among anecephalus, low- and high-level SB.

China ; epidemiology ; Female ; Humans ; Incidence ; Male ; Neural Tube Defects ; epidemiology ; Seasons