OBJECTIVETo establish a simple and rapid method to detect Streptococcus mutans and streptococcus sobrinus simultaneously in human saliva.

METHODSChromosomal DNA from the bacteria was obtained by the extraction method with phenol-chloroform. A nested PCR method with two sets of primers specific for portions of the glucosyltransferase genes (gtfB of S. mutans and gtfI of S. sobrinus), was optimized to detect S. mutans and S. sobrinus from standard strains, clinical strains and directly in human saliva.

RESULTSThe first process of nested PCR was capable of amplifying DNA fragments specific for these species from chromosomal DNA extracted from 10(5) CFU cells of standard and clinical strains, or from 1 ml clinical saliva samples containing 10(5) CFU cells of either species. a second process of nested PCR, using the first PCR product as a template with new internal primers to detect 10(3) CFU of either streptococcal species in 1ml saliva samples.

CONCLUSIONNested PCR could detect S. mutans and S. sobrinus rapidly and simply in human saliva. This finding would be important to studies of elucidation the role of these two streptococcal species in the etiology of dental caries.

Humans ; Polymerase Chain Reaction ; Saliva ; microbiology ; Streptococcus mutans ; genetics ; isolation & purification ; Streptococcus sobrinus ; genetics ; isolation & purification

This study was aimed to investigate the clinical feasibility of using multiplex PT-PCR assay for screening rare/cryptic chromosome translocations in patients with de novo acute myeloid leukemia. For 126 patients with de novo AML-M4/M5 without common chromosome translocations including t(15;17), t(8;21) and t(16;16), 3 parallel multiplex RT-PCR assays were set up to detect 6 mll-related gene rearrangements (mll/af10, mll/af17, mll/ell, mll/af9, mll/af6 and mll/enl) with low detection rate and 4 rare fusion genes (dek/can, tls/erg, aml1/mds (evi1) and npm/mlf1). The results showed that 11 patients with positive result from 126 patients were detected which involved in 5 molecular abnormalities. Among them, 10 cases were AML-M5 (16.67%), 1 cases AML-M4 (1.51%). The marker chromosomes were observed in 2 cases out of 11 cases through conventional karyotyping analysis, the karyotyping analysis in 1 case was not performed because this case had 1 mitotic figure only, no any cytogenetic aberrations were found in other 8 cases through R-band karyotyping analysis. It is concluded that multiplex RT-PCR designed in this study can quickly, effectively and accurately identify the rare/cryptic chromosome translocations and can be used in clinical detection.
Chromosome Banding ; Gene Rearrangement ; Genetic Testing ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Translocation, Genetic

OBJECTIVEThe effect of vascular endothelial growth factor(165) (VEGF(165)) on intracellular free magnesium ([Mg(2+)](i)) and the relationship between Mg(2+) and angiogenesis in human umbilical vein endothelial cells (HUVECs) were investigated in this study.

METHODS[Mg(2+)](i) in HUVECs loaded with fluorescent magnesium indicator mag-fura-2 were quantitatively detected with the use of intracellular cation measurement system. HUVECs were obtained from normal fetus and cultured in M199 with 0.2 fetal bovine serum. The angiogenesis effects of VEGF(165) were observed in presence of 0 mmol/L, 1 mmol/L or 2 mmol/L of extracellular Mg(2+).

RESULTSVEGF(165) significantly increased [Mg(2+)](i) in a dose-dependent manner independent of extracellular Mg(2+), Na(+) and Ca(2+) and this effect could be blocked by pretreatment with VEGF(165) receptor-2 (KDR) inhibitor (SU1498). The angiogenesis induced by VEGF(165) was significantly inhibited cells with 0 mmol/L extracellular Mg(2+), the angiogenesis effects of VEGF(165) were similar in cells with 1 mmol/L and 2 mmol/L extracellular Mg(2+) and these effects could be blocked by SU1498.

CONCLUSIONSThese results suggest that the [Mg(2+)](i) increase induced by VEGF(165) originates from intracellular Mg(2+) pools and promotes angiogenesis via KDR-dependent signaling pathways.

Cations, Divalent ; Cells, Cultured ; Endothelial Cells ; metabolism ; Humans ; Magnesium ; metabolism ; Neovascularization, Physiologic ; Signal Transduction ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Objective To study the routine examination of Aconitum sinomontanum Nakai and determine the contents of lappacontine and ranaconitine; To provide basis for establishing the quality standard of Aconitum sinomontanum Nakai.Methods Aconitum sinomontanum Nakai were collected from different areas.A method of TLC was used for qualitative discrimination. The methods in the Chinese Pharmacopoeia were adopted for the determination of moisture content, ash content and extractives. Determination of lappacontine and ranaconitine were performed by HPLC. Results The TLC showed that the spots were clear and the separation was good. Individual provisional standards:the moisture,total ash and acid-insoluble ash content of Aconitum sinomontanum Nakai were not more than 11.0%, 12.0%, and 7.0%, respectively; water soluble and alcohol soluble extractives were not less than 18.2% and 10.6%,respectively.The content of ranaconitine and lappacontine in Aconitum sinomontanum Nakai were not less than 0.125% and 0.815%, respectively. Conclusion The method established by the study is accurate and reliable,and can be used for quality evaluation of Aconitum sinomontanum Nakai.

Objective To investigate the distribution and antimicrobial resistance of pathogens causing bacterial peritonitis,provide laboratorial guidance for rational use of antimicrobial agents.Methods Pathogenic strains iso-lated from peritoneal fluid specimen of patients with peritonitis in the Affiliated Hospital of Xuzhou Medical Univer-sity in 2011-2015 were collected,performed bacterial identification and antimicrobial susceptibility testing,distri-bution of pathogens and antimicrobial resistance were analyzed.Results A total of 491 strains were collected,in-cluding 291(59.26%)strains of gram-negative bacilli,196(39.92%)of gram-positive cocci,and 4 (0.82%)of fun-gi.The top 5 pathogens were Escherichia coli (30.14%),coagulase negative staphylococcus(12.22%),Staphylo-coccus aureus (10.39%),Klebsiella pneumoniae (8.55%),and Enterococcus faecium(6.52%).Antimicrobial re-sistance rates of Escherichia coli ,Klebsiella pneumoniae ,Acinetobacter baumannii ,and Pseudomonas aeruginosa to imipenem were 4.90%,31.04%,77.28% and 26.27% respectively.Methicillin-resistant Staphylococcus aureus (MRSA)and methicillin-resistant coagulase negative staphylococcus(MRNCS)accounted for 56.02% and 70.02%respectively.Conclusion The main pathogens causing bacterial peritonitis are gram-negative bacilli,Escherichia co-li ranks first;resistance of pathogens is serious,standard use of antimicrobial agents should be strengthened to re-duce the emergence of drug-resistant strains.

OBJECTIVETo study the clinical efficacy of the frog shape four-step pulling method for the treatment of sacroiliac joint subluxation, and to explore the rationality of the method.

METHODSAmong 92 patients diagnosed as sacroiliac joint subluxation, 19 patients were male and 73 patients were female,ranging in age from 14 to 61 years, with an average of 38.7 years. The course of disease ranged from 1 month to 96 months,with an average of 18.4 months. All the patients were treated with the frog shape four-step pulling method. The therapeutic effects were observed.

RESULTSThe treatment times ranged from 1 to 10 (averaged 4.3 times). According to evaluation standard of sacroiliac joint subluxation, 71 patients got an excellent result, 19 good, 1 fair and 1 bad.

CONCLUSIONThe frog shape four-step pulling method had many advantages such as simplicity of operation, targeted, high rate of success, not easy rebound and less pain for patients and so on.

Adolescent ; Adult ; Female ; Humans ; Joint Dislocations ; therapy ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Sacroiliac Joint ; injuries

Objective:To observe the clinical effect of lower-reinforcing and upper-reducing acupuncture method for hyperplasia of mammary gland (HMG) and its influence on estradiol (E2) and progesterone (P).Methods:A randomized,single-blinded and controlled trial was conducted.A total of 124 cases conforming to the inclusion criteria were randomized by random number table into a treatment group and a control group,with 62 cases in each group.Patients in both groups received acupuncture therapy at the same acupoints,while patients in the treatment group received lower-reinforcing and upper-reducing method,and patients in the control group received even reinforcing-reducing manipulation.The treatment started around 10 d before menstruation and was conducted every other day.Patients received 5 treatments in each menstruation cycle for consecutive 3 cycles.The levels of E2,P and E2/P and clinical efficacy were measured before and after treatment.Results:After treatment,the breast lump size,pain intensity and concomitant symptoms score in both groups were substantially lower than those before treatment,showing statistical significances (all P＜0.01),and the improvement in the treatment group was higher than that in the control group,and the between-group comparisons showed statistical significances (all P＜0.01).After treatment,the overall effective rate was 91.9％ in the treatment group,higher than 72.6％ in the control group,and the between-group comparison showed a statistical significance (P＜0.01).After treatment,levels of E2,P and E2/P value showed no statistical significance when compared with those before treatment in the two groups (all P＞0.05).Conclusion:Lower-reinforcing and upper-reducing acupuncture method can effectively alleviate clinical symptoms and signs in HMG patients,and produce a better effect than even reinforcing-reducing manipulation.The majority of HMG patients' E2,P level and E2/P value were not beyond the normal ranges;therefore,acupuncture showed no substantial influence on E2 and P levels and E2/P value.

OBJECTIVETo study the expression level of cyclin D1-CDK4 protein in human embryonic lung fibroblasts (HELF) induced by quartz, and to study whether the expression level of cyclin D1-CDK4 protein mediated by mitogen activated protein kinase (MAPK)/(AP-1) signaling pathways.

METHODSCells were harvested after stimulation 2 h for the detection of cytokines. Cyclin D1 and CDK4 (cyclin dependent kinase 4) proteins was measured by immunocytochemistry (IC) and Western blot (WB).

RESULTSThe exposure of HELF to crystalline quartz for 2 hours could cause the decrease of cyclin D1 and cyclin dependent kinase 4 (CDK4) protein expression level, (7.91 +/- 0.29) x 10(3) and (5.17 +/- 0.28) x 10(4) respectively, which was lower than that of the HELF group (P < 0.05). AG126 (chemical inhibitor of the extracellular signal regulated protein kinase (ERK) signaling pathway) and the dominant negative mutant of ERK2 (molecular inhibitor of ERK2), prevented the decrease of cyclin D1 and CDK4 protein expression level. The chemical inhibitor of c-Jun NH2-terminal amino kinase (JNK), SP600125, could prevent both cyclin D1 and CDK4 protein expression level decrease. But SB203580, the chemical inhibitor of p38, prevented neither cyclin D1 nor CDK4 protein expression level decrease. Curcumin could prevent CDK4 protein expression level decrease but not cyclin D1 protein.

CONCLUSIONERKs and JNKs, but not p38, are responsible for the decrease of cyclin D1 and CDK4 protein expression level in HELF induced by quartz. AP-1 is responsible for the decrease of CDK4 expression level but not that of cyclin D1.

Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Humans ; Lung ; cytology ; embryology ; Mitogen-Activated Protein Kinases ; metabolism ; Quartz ; toxicity ; Transcription Factor AP-1 ; metabolism

OBJECTIVETo analyze retrospectively the clinical outcome of surgical management for upper cervical spine injury caused by trauma.

METHODSFrom January 2005 to March 2007, 16 patients with injury of upper cervical spine were treated by different management. There were 11 males and 5 females with an average age of 44 years ranging from 24 to 75. Of all, 5 cases were the odontoid fracture, 3 were atlas fracture, 5 were Hangman's fracture, 3 were atlanto-axial dislocation. MR imaging of cervical spine showed cervical cord compression and changes of T2 high signal in 5 cases. According to the injury mechanism, the imageological appearance, fracture classification, the methods of treatment were selected.

RESULTSSeven patients received non-operative treatment and nine patients underwent operation. Sixteen patients were followed up for 7 to 34 months (means 10.5 months). All fractures were healing or bone graft fusion and no internal fixation was lossing. There were no injuries of vertebral artery, nerve root or spinal cord.

CONCLUSIONCT and MRI are required in the course of diagnosis for the traumatic injury of upper cervical spine. The optimal modus operandi should be choose to retain upper cervical spine, meanwhile, can reserve the cervical movement.

Adult ; Aged ; Cervical Vertebrae ; injuries ; Female ; Fracture Healing ; physiology ; Humans ; Joint Instability ; Male ; Middle Aged ; Spinal Cord Injuries ; surgery ; Spinal Fractures ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVE To establish an UPLC-MS/MS method for the determination of lithospermoside and griffonilide in rat plasma and study its pharmacokinetic characteristics.METHODS The separation was performed on Aglient C18 column (100 mm × 2.1 mm,1.8 μm) with acetonitrile (A) and 0.1％ aqueous formic acid (B) as the gradient elution mobile phase (0 ～1 min,5％～20％B;1～2min,20％～60％B;2～6min,60％～70％B;6～6.Smin,70％～5％B;6.5～7.5min,5％B),the flow rate was 0.3 mL/min,the column temperature was set at 35 ℃ and the injection volume was 3 μL.UPLC-MS/MS was used to detected the concentration of lithospermoside and griffonilide in rat plasma after oral administration of Semiaquilegiae Radix.The pharmacokinetics parameters of lithospermoside and griffonilide were calculated by DAS 2.0 software.RESULTS Good linearity was found in lithospermoside and griffonilide within linear range from 0.701 9 ～ 140.4 ng/mL and 1.120 2 ～ 240.4 ng/mL,respectively.The RSD of accuracy and intra-/inter-day precision were less than 15％.The recovery of the method was ranged from 87.22 ％ to 107.75 ％.Lithospermoside and griffonilide were absorbed rapidly,the highest plasma concentration reached at 0.5 h,and the drug removed from body very fast and eliminated completely in 24 h.CONCLUSION This method is accuracy and stability for the study on pharmacokinetics of lithospermoside and griffonilide in rat plasma.