OBJECTIVE: To establish a 15-plex rapid STR multiplex amplification system. METHODS: Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives. RESULTS: Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure. CONCLUSION The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.
Alleles ; Chromosome Mapping ; DNA/genetics* ; DNA Fingerprinting/methods* ; Forensic Genetics/methods* ; Humans ; Microsatellite Repeats/genetics* ; Polymerase Chain Reaction/methods* ; Racial Groups/genetics* ; Sensitivity and Specificity ; Tandem Repeat Sequences

OBJECTIVE:To provide reference for rational application of Levofloxacin hydrochloride injection in the clinic. METHODS:With reference to the package insert of Levofloxacin hydrochloride injection,the guiding principles of clinical use of antibiotics,by reviewing related literatures,based on the weighted TOPSIS methods,detailed rules for drug utilization review (DUR) of Levofloxacin hydrochloride injection were made. And then 100 archived medical records of Levofloxacin hydrochloride injection in the first half of 2014 were evaluated in respect of medication rationality based on these rules. RESULTS:Among 100 cases,relative proximity of 51 cases was more than 70%(51.0%);that of 37 was between 50%-70%(37.0%);that of 12 cases was between 30%-50%(12.0%). CONCLUSIONS:Established DUR method of Levofloxacin hydrochloride injection on the basis of weighted TOPSIS methods can be used to evaluate the rationality of drug use and promote more rational evaluation behavior. And the results indicate that unreasonable use of Levofloxacin hydrochloride injection is still common in the hospital.

BACKGROUNDVisfatin, a visceral fat-derived adipocytokine, plays a significant physiological function in lipid metabolism. However, the precise function of visfatin and its regulation by thyroid hormones are still unknown. This study observed the plasma visfatin concentrations in subjects with hyperthyroidism and hypothyroidism in vivo, and investigated the possible regulation mechanism between visfatin and tri-iodothyronine (T3) in vitro as a further interpretation.

METHODSThe experiment in vivo included clinical subjects (57 patients with thyroid dysfunction and 29 euthyroid healthy volunteers) and an animal model (24 Wistar rats). All subjects were divided into hyperthyroidism, hypothyroidism and euthyroidism groups, with plasma thyroid hormones, thyrotropin, visfatin and triglyceride concentrations determined. Visfatin mRNA expression in visceral fat and liver of rats was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). The experiment in vitro studied 3T3-L1 cells and visfatin mRNA expression under nine different T3 concentrations (0, 0.1, 0.25, 0.5, 1, 5, 10, 20, 100 nmol/L) using quantitative real-time RT-PCR.

RESULTSClinical subjects and animal models showed elevated plasma visfatin concentrations in the hyperthyroidism group (20.466 ng/ml (15.263, 26.795 ng/ml) and (1209.164±165.292) ng/L) and hypothyroidism group (12.457 ng/ml (11.115, 15.454 ng/ml) and (1205.425±109.200) ng/L) compared to euthyroidism group (6.891 ng/ml (5.888, 8.803 ng/ml) and (926.650±54.002) ng/L, P<0.001). For animal models, visfatin mRNA expression in visceral fat in the hyperthyroidism and hypothyroidism groups increased about 3.33-fold and 1.98-fold compared to the euthyroidism group (P<0.001), which was positively correlated with plasma visfatin concentrations (r=0.713, P<0.001). However, no significant group difference (P>0.05) and correlation (r=0.121, P=0.572) was found in the liver. T3 induced a remarkable increase of visfatin mRNA expression in 3T3-L1 cells at low concentrations (0-0.5 nmol/L T3) followed by a sharp decrease at higher concentrations (0.5-100 nmol/L T3), with an inflection point at 0.5 nmol/L T3.

CONCLUSIONElevated circulating visfatin levels in subjects with hyperthyroidism and hypothyroidism are possibly due to an increase of visfatin mRNA expression in visceral fat, and a nonlinear regulation mechanism on visfatin mRNA expression under various T3 concentrations might be involved.

3T3-L1 Cells ; Adult ; Animals ; Female ; Humans ; Hyperthyroidism ; blood ; genetics ; metabolism ; Hypothyroidism ; blood ; genetics ; metabolism ; Male ; Mice ; Middle Aged ; Nicotinamide Phosphoribosyltransferase ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Triiodothyronine ; blood

Objective To study characteristics of rpoB gene mutations associated with Mycobacterium tuberculosis( MTB) resistance to rifampicin in Guizhou of China to discuss rapid screening methods of detecting MTB resistance to rifampicin.Methods MTB DNA of clinical samples were tested by fluorescent PCR .Low DNA concentration samples were excep-ted.All MTB DNA positive samples were tested by gene chip to screen rpoB gene mutation.The samples tested to be abnormal were further sequenced and aligned. Results MTB DNA of 3966 samples was positive.The positive rate was 22.72%(3966/17452 ).All of the 2738 samples were tested by gene chip , of which 552 samples were detected to have gene mutations and the rate was 20.16%( 552/2738 ) .Nine sam-ples of rpoB gene were tested abnormal .These 9 samples were confirmed to have nucleotide deletion , insertion, the same codon mutation of double base or multisite mutation .There were 26 types of gene mutations and mainly were at site 531 , site 526 , site 516 , site 511 , site 533 , site 513 and site 512 and the mutation rates were 56.23%, 18.09%, 9.67%, 8.61%, 4.57%, 1.76% and 0.18%, separately.Conclusion The characteristics of rpoB gene mutations associated with MTB resistance to rifampicin in Guizhou were basically the same as those in the literature reports.The gene chip can be used as the rapid screening method to detect MTB resistance to rifampicin in Guizhou .

Objective To explore the relationship between duration of sleeping and cerebral infarction.Methods A case-control study involved 1037 cerebral infarction patients admitted by the Second Affiliated Hospital of Harbin Medical University,December 2011-December 2012 as cases.Another 1205 adults free from cerebro-vascular diseases who had undergone physical examination in the hospital at the same period,were served as controls.All the subjects were interviewed with unified questionnaire.Chi-square test,u-test and multivariate logistic regression analysis were performed.Results After adjustment for potential confounding factors including age,sex,body mass index,wrist-hip ratio,smoking,alcohol intake,hypertension,diabetes mellitus,coronary artery disease and lipid parameters,data from the multivariate logistic regression analysis showed that the risk of cerebral infarction was greater in people who slept less than 6 hours per night than those who slept between 6 hours and 8 hours per night,with an odds ratio (95％ CI) as 2.81 (95％ CI:1.68-4.70).There was no significant association between factor as ‘sleeping longer than 8 hours/pre day' and cerebral infarction.Through the subgroup analysis,data showed that the association between ‘ shorter than 6 hour sleep/night' and cerebral infarction consistently exsited,across the categories of sex,and the degree of association was greater in women than in men,with the odds ratio as 5.58 (95％CI:1.78-17.52) and 2.00(95％CI:1.10-3.64) respectively.Conclusion Short sleeping duration might increase the risk of developing cerebral infarction.

OBJECTIVETo investigate the significance of flexible bronchoscopy in children with respiratory diseases.

METHODSA retrospective analysis was performed for the clinical data of 80 children who were hospitalized due to respiratory diseases (including severe pneumonia, Mycoplasma pneumoniae pneumonia with atelectasis/lung consolidation/local emphysema, protracted pneumonia, coughing and wheezing of unknown cause, chronic cough of unknown cause, and laryngeal stridor) and who underwent flexible bronchoscopy/alveolar lavage.

RESULTSBronchoscopy found that all the 80 children had endobronchial inflammation, among whom 28 children had severe airway obstruction by secretion. Twenty-four children had congenital airway dysplasia besides endobronchial inflammation, and three children had bronchial foreign bodies. In the children with coughing and wheezing of unknown cause and laryngeal stridor, some had congenital airway dysplasia or bronchial foreign bodies. Among the 27 children with Mycoplasma pneumoniae pneumonia, 26 had severe airway obstruction/embolization by secretion; 25 children (93%) underwent chest imaging again at 2 weeks after alveolar lavage, and the results showed complete or partial lung recruitment. Among the 80 children who underwent bronchoscopy, 3 had severe hypoxemia during surgery, 1 had epistaxis, 1 had minor bleeding during alveolar lavage, 3 had transient bronchospasm, and 5 had postoperative fever; these children were all improved after symptomatic treatment.

CONCLUSIONSFlexible bronchoscopy is safe and reliable in children with respiratory diseases. Early alveolar lavage under a flexible bronchoscope is recommended for children with severe/refractory Mycoplasma pneumoniae pneumonia to improve prognosis. Flexible bronchoscopy is recommended for children with recurrent coughing and wheezing and persistent laryngeal stridor, in order to directly observe the throat and airway under an endoscope.

Bronchoscopy ; adverse effects ; methods ; Child ; Child, Preschool ; Cough ; diagnosis ; Female ; Humans ; Infant ; Lung Diseases ; diagnosis ; Male ; Pneumonia, Mycoplasma ; diagnosis ; Respiratory Sounds ; diagnosis ; Retrospective Studies

OBJECTIVETo evaluate the clinical effect of a 10-day sequential therapy which was made up of omeprazole, clarithromycin, amoxicillin-clavulanate and metronidazole for the eradication of Helicobacter pylori (Hp) infection in children.

METHODA total of 214 children with abdominal pain, who were confirmed to have Hp infection through endoscopy, biopsy, and Hp culture. The 214 cases were randomly divided into four groups. A 10-day sequential therapy group accepted omeprazole 0.8 - 1.0 mg/(kg·d) plus amoxicillin-clavulanate 50 mg/(kg·d) for five days and omeprazole 0.8 - 1.0 mg/(kg·d), clarithromycin 20 mg/(kg·d) and metronidazole 20 mg/(kg·d) for the remaining five days. The 7-day triple therapy group, 10-day triple therapy group and 14-day triple therapy group received omeprazole 0.8 - 1.0 mg/(kg·d), amoxicillin-clavulanate 50 mg/(kg·d) and clarithromycin 20 mg/(kg·d) for 7 days,10 days,14 days, respectively. All drugs were given twice daily. All these patients received (13)C urea breath test ((13)C-UBT) four weeks after the treatment.

RESULTFinally, 199 patients were followed up, and the total rate of loss to follow-up was 7.0% (15/214). Hp eradication rate was 85.2% and 90.2% in the 10-day sequential therapy group on intention to treat (ITT) and per protocol (PP) analyses, 66.0% and 71.4% in the 7-day triple therapy group on ITT and PP analyses; 60.0% and 67.3% in 10-day triple therapy group on ITT and PP analyses, and 78.8% and 82.0% in patients who received the 10-day sequential regimen on ITT and PP analyses, respectively. By ITT analysis, there was significantly difference between the 10-day sequential therapy group and 7-day or 10-day triple therapy group (P < 0.05), while no significant difference was found between the 10-day sequential therapy group and 14-day triple therapy group (P > 0.05). The results of the ITT analysis and the PP analysis were the same. The four groups had neither significant difference in abdominal pain relief (P > 0.05) nor in incidence of adverse reactions (P > 0.05).

CONCLUSIONThe 10-day sequential regimen was significantly more effective than both 7-day triple regimen and 10-day triple regimen, while had the same eradication rate compared with the 14-day sequential therapy. But 10-day triple regimen to eradicate Hp infection in children had the advantages such as short course of treatment and better compliance.

Administration, Oral ; Adolescent ; Amoxicillin ; administration & dosage ; adverse effects ; Anti-Bacterial Agents ; administration & dosage ; adverse effects ; Anti-Ulcer Agents ; administration & dosage ; Breath Tests ; methods ; Child ; Child, Preschool ; Clarithromycin ; administration & dosage ; adverse effects ; Drug Administration Schedule ; Drug Therapy, Combination ; Female ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; drug effects ; isolation & purification ; Humans ; Male ; Metronidazole ; administration & dosage ; adverse effects ; Microbial Sensitivity Tests ; Omeprazole ; administration & dosage ; adverse effects ; Time Factors ; Treatment Outcome

OBJECTIVETo study clinical features, treatment and curative effects in children with acute clenbuterol poisoning, in order to provide a basis for early diagnosis and treatment.

METHODSClinical data of 28 hospitalized children with acute clenbuterol poisoning in April 2011 were retrospectively studied.

RESULTSOf the 28 patients, there were 15 males and 13 females, aged 1 to 13 years (mean age 6.5±4.8 years). Vomiting, palpitations and limb shaking were found as main clinical manifestations in the patients. Main changes of blood biochemical included hypokalemia, lactic acidosis, hyperglycemia, hypsocreatinkinase. Snus tachycardia and S-T segment depression were observed on ECG. Patients' symptoms were gradually alleviated after 12-78 hours by use of beta blockers, potassium supplement, protecting the heart and other symptomatic and supportive treatment. Blood biochemical indexes were improved after 48 hours of admission. All of the patients were cured after 5 days. The symptoms of the patients do not longer occur during a follow up of half a month.

CONCLUSIONSAcute clenbuterol poisoning is characterized by vomiting, palpitations, limb shaking, hypokalemia, lactic acidosis and tachycardia in children. An early effective treatment of this disease can improve prognosis in children.

Acute Disease ; Adolescent ; Adrenergic beta-Agonists ; poisoning ; Child ; Child, Preschool ; Clenbuterol ; poisoning ; Electrocardiography ; Female ; Humans ; Infant ; Male ; Retrospective Studies

OBJECTIVETo investigate the significance of CD11b expression in neutrophils and lymphocytes in children with systemic inflammatory response syndrome (SIRS).

METHODSCD11b expression in neutrophils and lymphocytes was measured using flow cytometry in 36 children with SIRS (SIRS group) and 28 children with infectious disease but without SIRS (control group). The sensitivity and specificity of neutrophil CD11b for diagnosis of SIRS were evaluated.

RESULTSDuring the acute phase, an increased CD11b expression in neutrophils (96.7+/-8.1%) was observed in the SIRS group compared with the control group (85.1+/-5.1%) (p<0.05). Using neutrophil CD11b expression >92.2% as a cut-off value for diagnosis of SIRS, the sensitivity and the specificity were 97.2 % and 92.9% respectively. Lymphocytic CD11b expression in the SIRS group (13.4+/-8.6%) was lower than that in the control group (19.2+/-6.4%) in the acute phase (p<0.05). In the SIRS group, lymphocytic CD11b expression was remarkably suppressed in the severe sepsis subgroup (7.27+/-3.04%), showing significantly decreased expression compared with the non-infectious subgroup (19.3+/-2.9%) and the sepsis subgroup (15.9+/-12.5%) (p<0.01). In the convalescence stage lymphocytic CD11b expression in the SIRS group was similar to that in the control group.

CONCLUSIONSCD11b expression in neutrophils may serve as a reliable indicator for diagnosis of SIRS. The down-regulation of lymphocytic CD11b expression might be a signal of the condition aggravation in children with SIRS.

C-Reactive Protein ; analysis ; CD11b Antigen ; blood ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Lymphocytes ; chemistry ; Male ; Neutrophils ; chemistry ; Sensitivity and Specificity ; Systemic Inflammatory Response Syndrome ; diagnosis ; immunology

Objective To investigate the infection rate of human papillomavirus(HPV)among pregnant women, and to explore the effect of HPV infection on adverse pregnancy outcome. Methods A total of 1 679 pregnant women in hospital were collected for the research. The flow-through hybridization and genechip(HybriMax)method was used to detect the infection of HPV. Univariate analysis was used to analyze the factors affecting HPV infection in pregnant women. The binary logistic analysis was used to analyze risk factors affecting adverse pregnancy outcome. Results HPV infection rate was 31.39%(527/1 679), including 14.23%(239/1 679)of HR-HPV, 15.54%(261/1 679)of LR-HPV and 1.61%(27/1 679)of mixed of HR-HPV and LR-HPV. Univariate analysis showed that there was significant difference in initial sex age, education level and smoking history between infection group and non-infection group, with statistical difference(P<0.05). The incidence rate of adverse pregnancy outcomes in infection group(31.50%) was significant higher than that of non-infection group(9.81%), with statistical difference(P<0.01). The incidence rate of premature rupture of fetal membranes, newborn respiratory papillomatosis and other adverse pregnancy outcomes among HR-HPV group, LR-HPV group and mixed group had no significant difference (P>0.05). Binary logistic regression analysis showed HR-HPV infection(OR=4.194, 95% CI: 3.099-5.675), LR-HPV infection(OR=1.771, 95%CI: 1.288-2.434)and mixed type infection(OR= 3.350, 95%CI: 1.630-7.735)were the risk factors affecting adverse pregnancy outcome(P<0.01), however, age and times of gestation had no statistical significance in the binary logistic analysis(P>0.05). Conclusion HPV infection was the risk factors for adverse pregnancy outcome, which indicated that screening work in pre-pregnancy and pregnancy, and persisting in early prevention, early detection and early treatment could reduce the incidence rate of adverse pregnancy outcome.