Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

OBJECTIVETo summarize early diagnosis and treatment methods of 20 patients with compartment syndrome caused by landslides during coal mine accidents in order to improve the level of diagnosis and treatment of compartment syndrome and reduce disability.

METHODSFrom September 2006 to April 2010,20 patients with compartment syndrome were treated with the methods of early decompression, systemic support. All the patients were male with an average age of 42 years (ranged, 23 to 54). All the patients with high tension limb swelling, pain, referred pain passive positive; 5 extremities feeling diminish or disappear and the distal blood vessel beat were normal or weakened or disappeared; myoglobinuria, hyperkalemia, serum urea nitrogen and creatinine increased in 5 cases and oliguria in occurred 1 case. The function of affected limbs was observed according to disability ratings.

RESULTSThree cases complicated with infection of affected limb and 6 cases occurred with renal function insufficiency. Total recovery was in 16 cases, basically recovery in 3, amputation in 1 case. All patients were followed up for 6-15 months with an average of 12 months. The ability to work according to national standard identification--Employee work-related injuries and occupational disability rating classification (GB/T16180-2006) to assess, grade 5 was in 1 case, grade 8 in 2 cases, grade 10 in 1 case, no grade in 16 cases.

CONCLUSIONArteriopalmus of dorsalis pedis weaken and vanished can not be regard as an evidence in early diagnosis of compartment syndrome. Early diagnosis and decompression, systemic support and treatment is the key in reducing disability.

Adult ; Compartment Syndromes ; diagnosis ; surgery ; Decompression, Surgical ; methods ; Early Diagnosis ; Humans ; Landslides ; Male ; Middle Aged ; Water-Electrolyte Imbalance ; therapy

AIMTo study unitary-core osmotic pump tablet for delivering water-insoluble drug for 24 hours.

METHODSUnitary-core osmotic pump tablet was prepared using nifedipine as the model drug. The effects of various core formulation variables on drug release were studied.

RESULTSPolyethylene oxide and potassium chloride have comparable positive effects on drug release, whereas, nifedipine has markedly negative effect on drug release.

CONCLUSIONUnitary-core osmotic pump tablet is very easy in preparation and it can deliver water-insoluble drug in substantially constant rate for 24 hours.

Calcium Channel Blockers ; administration & dosage ; chemistry ; Delayed-Action Preparations ; Drug Delivery Systems ; Nifedipine ; administration & dosage ; chemistry ; Osmosis ; Polyethylene Glycols ; pharmacology ; Potassium Chloride ; pharmacology ; Solubility ; Tablets ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the sleep quality and related factors among medical students in China, understand the association between dormitory environment and sleep quality, and provide evidence and recommendations for sleep hygiene intervention.

METHODSA total of 555 undergraduate students were selected from a medical school of an university in Beijing through stratified-cluster random-sampling to conduct a questionnaire survey by using Chinese version of Pittsburgh Sleep Quality Index (PSQI) and self-designed questionnaire. Analyses were performed by using multiple logistic regression model as well as multilevel linear regression model.

RESULTSThe prevalence of sleep disorder was 29.1%(149/512), and 39.1%(200/512) of the students reported that the sleep quality was influenced by dormitory environment. PSQI score was negatively correlated with self-reported rating of dormitory environment (γs=-0.310, P<0.001). Logistic regression analysis showed the related factors of sleep disorder included grade, sleep regularity, self-rated health status, pressures of school work and employment, as well as dormitory environment. RESULTS of multilevel regression analysis also indicated that perception on dormitory environment (individual level) was associated with sleep quality with the dormitory level random effects under control (b=-0.619, P<0.001).

CONCLUSIONSThe prevalence of sleep disorder was high in medical students, which was associated with multiple factors. Dormitory environment should be taken into consideration when the interventions are taken to improve the sleep quality of students.

Beijing ; epidemiology ; Housing ; Humans ; Logistic Models ; Prevalence ; Risk Factors ; Schools, Medical ; Self Report ; Sleep Wake Disorders ; epidemiology ; Social Environment ; Students, Medical ; psychology ; statistics & numerical data

OBJECTIVETo investigate body weight and the body mass index (BMI) in girls with adolescent idiopathic scoliosis (AIS).

METHODSSix hundred and thirteen AIS female patients admitted from January 2005 to January 2007 and 449 healthy girls were recruited for this study with a range of age from 12 to 16 years old. The direct indexes included body height, body weight and date of birth. The indirect indexes included corrected body height, BMI and corrected BMI. The corrected body heights of AIS patients were adjusted by using the greatest Cobb angle to correct for spinal deformity according to Bjure's formula. Two-tailed Student's test was used for group comparison.

RESULTSIn the AIS patients, the maximum Cobb angle was (31 +/- 11) degrees (11 degrees to 77 degrees). The corrected body height were significantly higher in AIS patients than in the normal controls from 12 to 16 years old (P < 0.05). The body weight were significantly lower in AIS girls than in normal controls from 12 to 16 years of age (P < 0.01). From 12 to 16 years of age, the BMI were (17.6 +/- 1.9), (17.9 +/- 2.5), (17.9 +/- 2.1), (18.6 +/- 2.3) and (19.0 +/- 1.9) kg/m2 in AIS patients; while the BMI were (19.5 +/- 3.4), (19.8 +/- 3.0), (20.4 +/- 2.9), (20.4 +/- 2.8) and (20.2 +/- 2.2) kg/m2 in normal controls. The BMI were significantly lower in AIS girls than the normal controls from 12 to 16 years of age (P < 0.001).

CONCLUSIONSAfter the onset of puberty, significantly longer corrected height, lower body weight and lower BMI were found in AIS patients. Results of this large-scale study revealed the presence of abnormal growth in AIS patients during peripubertal development.

Adolescent ; Body Height ; Body Mass Index ; Body Weight ; Child ; Female ; Humans ; Kyphosis ; physiopathology ; Scoliosis ; physiopathology

OBJECTIVETo explore the location of the anterior border of facets and the posterior border of vertebral bodies in lower cervical spine,and to provide a quantitative data to evaluate the correct length of transarticual screws in lower cervical spine during procedure.

METHODSOne hundred standard lateral X-ray films and fifty CT films on cervical spine were used to measure the distance of the anterior border of facets and the posterior border of vertebral bodies in lower cervical spine. HS, HM and HI were defined as parameters, which means the distance between the anterior border of the superior (HS), median (HM) and inferior (HI) part of facets and the posterior border of corresponding vertebral bodies. The value will be negative if the anterior border of the facet located before the vertebral body.

RESULTS'HS > HM > HI' was found in all facets in lower cervical spine. The anterior border of the facet in C(3,4) located before the posterior border of the vertebral body of C3. The anterior border of C(4,5) and C(5,6) was inclined to posterior. The anterior border of C(6,7) located after the posterior border of the vertebral body of C6. The pattern of HS increased from C(3,4) to C(6,7), the minimal (0 +/- 0.25) mm and the maximal (2.91 +/- 1.05) mm. The tendency of HM raised from C(3,4) to C(6,7), the minimal (-1.57 +/- 0.53) mm and the maximal (1.54 +/- 0.39) mm. The pattern HI added from C(3,4) to C(6,7), the minimal (-2.03 +/- 0.40) mm and the maximal (1.08 +/- 0.70) mm.

CONCLUSIONDuring the implantation of the transarticular screws, the tip of the screws should be 0-2 mm before the posterior border of the vertebral body of C3 at C(3,4), 0-2 mm after that of C4 at C(4,5), 0.5-2.5 mm at C(5,6) and 1-3 mm at C(6,7). The quantitative location between the anterior border of facets and the posterior border of the corresponding vertebral bodies can offer an indirect method to evaluate the correct length of transarticual screws in lower cervical spine during procedure.

Cervical Vertebrae ; chemistry ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Middle Aged ; Spinal Diseases ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Zygapophyseal Joint ; chemistry ; surgery

Objective: By observing the effects of electroacupuncture (EA) on the apoptosis of conjunctival cells of rabbits with dry eye syndrome (DES) and the expressions of apoptosis-related proteins Caspase-3, Fas and Bcl-2, to discuss the mechanism of EA in the treatment of DES from the perspective of cell apoptosis. Methods: Male New Zealand rabbits were randomly divided into a normal group (NG), a model group (MG), an EA group (EAG) and a sham EA group (SEAG). DES rabbit model was developed by eye drop of 0.1％ benzalkonium chloride. The rabbit tear secretion and tear film break-up time (BUT) were measured; terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling (TUNEL) assay was used to detect the apoptosis of conjunctival cells; the expressions of Caspase-3, Fas and Bcl-2 proteins in conjunctival cells were detected by immunohistochemistry. Results: Compared with the NG, the rabbit tear secretion decreased and the BUT was shortened in the MG (both P<0.01); compared with the MG and the SEAG, the rabbit tear secretion increased and the BUT was prolonged in the EAG (all P<0.05). Compared with the NG, the apoptosis of rabbit conjunctival cells increased (P<0.01), the expressions of Caspase-3 and Fas proteins increased (both P<0.05), and the expression of Bcl-2 protein decreased (P<0.01) in the MG; compared with the MG and the SEAG, the apoptosis of rabbit conjunctival cells decreased (both P<0.01), the expressions of Caspase-3 and Fas proteins decreased (all P<0.05), and the expression of Bcl-2 protein increased (both P<0.01) in the EAG. Conclusion: EA can inhibit the apoptosis of rabbit conjunctival cells, down-regulate the expressions of apoptosis-related proteins Caspase-3 and Fas, and up-regulate the expression of Bcl-2 protein, which may be one of the mechanisms of EA in treatment of DES.

The chitosan thermosensitive hydrogel is liquid at room temperature but gels rapidly when heated to body temperature. This hydrogel are wildly used for cell encapsulation, drug delivery or tissue-engineered scaffolds. The system can sustain the release of macromolecules over a period of several hours to a few days. However, with low-molecular-weight compounds, the release is generally completed within 24 h. To prepare a functional chitosan thermosensitive hydrogel for slow release both broad-spectrum antibiotic chlorhexidine and growth factor recombined human bone morphogenetic protein-2 (rhBMP-2), The beta-cyclodextrin was used to prepare an inclusion complex with chlorhexidine, and then the latter was incorporated into the chitosan thermosensitive hydrogel system. Simultaneously, rhBMP-2 was added into the hydrogel system. By HAAKE viscosity measuring instrument, we contrasted the viscoelastic properties of system with or without objective factors. And the in vitro release kinetics of chlorhexidine and rhBMP-2 was investigated by HPLC (high performance liquid chromatography) and ELISA (enzyme-linked immunosorbent assay) respectively. The results showed that the addition of chlorhexidine/beta-cyclodextrin inclusion complex to the thermosensitive solution did not change the gelling behavior of the thermosensitive system. Further, the in vitro release profiles demonstrated that the release rate of chlorhexidine and rhBMP-2 from hydrogel became slower, controlled delivery over at least 1 month. By first preparing chlorhexidine/beta-cyclodextrin inclusion complex, and then mixing the IC and rhBMP-2 into the chitosan thermosensitive hydrogel, a functional chitosan thermosensitive hydrogel system with ability of slow release both rhBMP-2 and chlorhexidine is successfully made.
Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; administration & dosage ; Chitosan ; chemistry ; Chlorhexidine ; administration & dosage ; Delayed-Action Preparations ; chemical synthesis ; Drug Carriers ; chemistry ; Drug Combinations ; Hydrogels ; chemistry ; Recombinant Proteins ; administration & dosage ; Temperature ; Transforming Growth Factor beta ; administration & dosage

OBJECTIVETo investigate the association between the tissue inhibitor of metalloproteinase-2 (TIMP-2) gene polymorphisms with the predisposition and disease severity of thoracic adolescent idiopathic scoliosis (AIS).

METHODSThree hundred and fifty-four female thoracic AIS patients treated from January 2007 to March 2009 and 210 healthy female who underwent health examination during March 2005 to June 2006 as normal controls were recruited in this study. One SNP-418G/C (rs8179090) in the promoter region were selected for TIMP-2 gene. PCR- RFLP was used for genotyping.

RESULTSNo significant differences of genotype and allele frequency distribution were found between AIS patients and normal controls (P > 0.05). In AIS patients, the frequency of C allele of the patients with the body mass index (BMI) < 17 kg/m(2) was significantly higher than those with the BMI > or = 17 kg/m(2) (P < 0.05), and the frequency of C allele of cases with the major Cobb angle > or = 40 degrees was significantly higher than that with Cobb angle < 40 degrees (P < 0.05). Among the patients who reached skeletal maturity without any interference of natural history, significantly higher average maximum Cobb angle was found in patients with GC and CC genotype compare with those with GG genotype.

CONCLUSIONSThe SNP-418G/C (rs8179090) in the promoter region of TIMP-2 gene may be associated with abnormal growth pattern and curve progression of thoracic AIS. TIMP-2 gene is a disease-modifier gene of thoracic AIS.

Adolescent ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Scoliosis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; genetics