Molecular technologies are sensitive, fast, and cheap in the study on microbial ecology. However, these methods do not provide information about morphology, number, and spatial distribution of the microorganisms. In contrast, Fluorescence in situ hybridization (FISH) combines the precision of molecular biology with the visual information from microscope, and permits visualization and identification of individual miciobial cells within their natural habitat. FISH not only allows the detection of slow growing microorganisms, but also of yet-to-be cultured. It is useful for many applications in diagnosis and assessment of the population structure of complex microbial communities, and is a powerful tool for molecular ecology studies in microbiology, In this paper, major techniques and progresses of FTSH were described. Its application in microbial ecology, as well as problems , pitfalls, and perspectives of FISH are discussed.

Environmental Monitoring ; Humans ; Occupational Diseases ; etiology ; prevention & control ; Occupational Health Services ; organization & administration ; Population Surveillance ; methods ; Risk Assessment ; Surveys and Questionnaires ; standards

OBJECTIVETo analyze the epidemiological characteristics of pediatric burn in the midwest region of Inner Mongolia and the related areas, and to provide reference for seeking pertinent measures of prevention and treatment.

METHODSMedical records of patients hospitalized in the 253rd Hospital of PLA, the 322nd Hospital of PLA, and the Inner Mongolia Autonomous Region Hospital from January 1996 to December 2010 were collected. Patients were divided into pediatric burn group with specific reason (group SF, with scald resulted from construction defect of Guo-lian-kang--a heatable brick bed linked to a cooking pot), and burn control group with other causes (group C) according to the main injury cause. Clinical data of patients in both groups, including general condition, family background, occurrence regularity, and outcome, were analyzed. The epidemiological trend of variation before and after taking preventive measures (1996 to 2001 and 2002 to 2010) was compared. Data were processed with chi-square test and rank sum test.

RESULTS(1) General condition: out of 16 595 pediatric burn patients, 15 816 cases (95.3%) suffered scald due to liquids with high temperature, and 779 cases (4.7%) suffered burns due to other causes. Patients in group SF (scald due to specific cause--Guo-lian-kang) accounted for 32.2% (5089/15 816) of the total suffered scald by liquids with high temperature, and 30.7% (5089/16 595) of all the inpatients the cause of burn was related to Guo-lian-kang (group SF). The patients in group SF admitted to the 322nd Hospital of PLA accounted for 34.2% of all the inpatients admitted to this hospital (1803/5267), more than the other two hospitals in this study. The number of patients in group C was 11 506, accounted for 69.3% of all the inpatients. The age of patients ranged from 8 months to 5 years in group SF and 1 month to 12 years in group C. The age of the majority of patients ranged from 1 to 3 years in both groups. The ratio of male to female was 2.1:1.0 in group SF and 1.4:1.0 in group C. The incidence of scald involving multiple body parts in group SF (3590 cases accounting for 70.5%) was obviously higher than that of group C (6311 cases accounting for 54.8%, χ(2) = 361.138, P < 0.01). In both group SF and group C, the incidence in different sites was ranked from high to low as follows: upper limbs, lower limbs, the head-face-neck region, and the trunk. The degree of injury in group SF was much more severe than that of group C (Z = 27.770, P < 0.01). The rate of patients without pre-hospital treatment was 31.2% (1588/5089) in group SF, which was obviously higher than that of group C (24.8%, 2857/11 506, χ(2) = 73.010, P < 0.01). The rate of patients treated with cryotherapy was obviously lower in group SF (14.7%, 747/5089) than in group C (19.6%, 2255/11 506, χ(2) = 57.636, P < 0.01). The rate of patients treated with delayed resuscitation (6 hours after injury) in group SF (31.5%, 1601/5089) was obviously higher than that of group C (7.8%, 897/11 506, χ(2) = 1545.234, P < 0.01). (2) Family background and occurrence regularity: in group SF, 67.3% (3424/5089) of the patients came from farming area, 22.1% (1123/5089) from villages and towns, and 10.7% (542/5089) from urban areas. In group C, 32.4% (3727/11 506) of the patients came from farming area, 48.4% (5570/11 506) from villages and towns, and 19.2% (2209/11 506) from urban areas. Most of the patients in group SF (77.8%, 3958/5089) were injured between October and March, while most of the patients in group C (58.2%, 6697/11 506) were injured between May and October. (3) Outcome and epidemiological variation: the cure rate of patients in group SF was 32.3% (1645/5089), which was obviously lower than that of group C (44.7%, 5143/11 506, χ(2) = 215.615, P < 0.01). The mortality of patients in group SF was 1.6% (79/5089), and it was obviously higher than that of group C (0.4%, 46/11 506, χ(2) = 62.700, P < 0.01). From 1996 to 2001, patients in group SF accounted for 42.5% (2213/5212), while patients in group C accounted for 57.5% (2999/5212) of the inpatients scalded by hot liquid. After taking preventive measures against injury due to Guo-lian-kang, incidence of scald injury in group SF was lowered to 27.1% (2876/10 604), while the incidence in group C remained at 72.9% (7728/10 604) of the inpatients with hot liquid scald from 2002 to 2010. The difference between the two periods was statistically significant (χ(2) = 376.695,P < 0.01).

CONCLUSIONSThe defect of construction of Guo-lian-kang is one of the main factors that lead to a high incidence of pediatric burn in the midwest of Inner Mongolia. Installation of a protective bannister between the cooking pot and the "kang (heatable brick bed)" can obviously reduce the incidence of scald injury. Special injury-causing factors, unprofessional pre-hospital treatment of the wound, delayed resuscitation after shock are the main causes of increasing mortality and disability, and they constitute the key targets of prevention and treatment of such injury in future.

Burns ; epidemiology ; Child ; Child, Preschool ; China ; Female ; Humans ; Infant ; Male

OBJECTIVETo investigate whether mammalian target of rapamycin (mTOR) kinase was abnormally activated in maldeveloped balloon cells and dysmorphic neurons of focal cortical dysplasia (FCD) with refractory epilepsy.

METHODSA total of 12 archival cases of FCD typeIIwith medically intractable epilepsy treated between 2008 and 2010 were retrieved. Perilesional brain tissue was used as control specimens (n = 8). The expression of phosphorylated p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was investigated by imunocytochemistry.

RESULTSThe expression of p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was found in meldeveloped balloon cells and dysmorphic neurons of FCD. A weak stain in a small amount of pyramid neurons was also found in the control group.

CONCLUSIONAbnormal activation of mTOR in maldeveloped balloon cells and dysmorphic neurons of FCD may be a key molecular mechanism underlying the histological changes and repeated seizures.

Adolescent ; Adult ; Brain Diseases ; metabolism ; pathology ; Child, Preschool ; Epilepsy ; metabolism ; pathology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Male ; Malformations of Cortical Development ; metabolism ; pathology ; Malformations of Cortical Development, Group I ; Nestin ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; TOR Serine-Threonine Kinases ; metabolism ; Young Adult

OBJECTIVETo explore the technique and clinical results of close reduction by manipulation and minimally invasive percutaneous osteosynthesis with intramedullary nail for the treatment of femur shaft fractures. methods: A retrospective study was conducted to analyze 96 patients with the femur shaft fractures who had been treated with close reduction by manipulation and minimally invasive percutaneous osteosynthesis with intramedullary nail. There were 67 males and 29 females. The average age of patients was 39 years old (ranging from 16 to 88). According to AO fracture classification for the femur shaft fractures,there were 29 cases of type A,46 type B,21 type C.

RESULTSAll the patients were followed up and the duration ranged from 12 to 24 months (averaged, 15 months). All the fractures showed union. The time required for the bony union ranged from 3 to 10 months (averaged,4 months). The clinical results were evaluated by Thorsen classification system. At the latest follow-up, 87 patients obtained excellent results, 7 good, 2 fair.

CONCLUSIONThis treatment method combines advantages of intramedullary nail with close manipulative reduction, so can get satisfactory clinical results for the treatment of femoral shaft fracture with minimal trauma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Nails ; Female ; Femoral Fractures ; diagnostic imaging ; surgery ; therapy ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; instrumentation ; Musculoskeletal Manipulations ; methods ; Retrospective Studies ; Tomography, X-Ray Computed ; Wound Closure Techniques ; Young Adult

OBJECTIVETo observe the effects of ginkgo flavone aglycone (GA) on oxidized low-density lipoprotein (ox-LDL) induced oxidative injury of human aortic endothelial cells (HAECs) and its mechanisms.

METHODSHAECs were in vitro cultured. Then they were divided into 6 groups, i.e., the vehicle control group, the ox-LDL group, the GA 30 mg/L group, the GA 60 mg/L group, the GA 90 mg/L group, and the Vit E group. The oxidative injury model was duplicated in the rest 5 groups by adding 150 mg/L ox-LDL except the vehicle control group. GA was added as intervention at corresponding dose to the GA 30 mg/L group, the GA 60 mg/L group, and the GA 90 mg/L group. Vit E at 200 micromol/L was administered to those in the Vit E group. The survival rate of HAECs was detected by MTT. The contents of reactive oxygen species (ROS) in HAECs were determined by CM-H2DCFDA fluorescent probe. The contents of NADPH oxidase were detected by ELISA. The levels of malondialdehyde (MDA) were measured by thiobarbituric acid (TBA) test. The contents of nitric oxide (NO) were determined by Griess reagent method. The contents of superoxide dismutase (SOD) were detected by xanthine oxidase method.

RESULTSCompared with the vehicle control group (100.00%), the cell survival rate in the ox-LDL group (70.68%) obviously decreased (P <0.05). The cell survival rate was 88. 95% in the VitE group, 83.25% in the GA 30 mg/L group, and 94.93% in the GA 60 mg/L group, obviously higher than that of the ox-LDL group (70.68%, P <0.05). The optimal effects were shown in the GA 60 mg/L group. Compared with the vehicle control group, the contents of ROS, MDA, and NADPH oxidase increased, the contents of NO and the SOD activity decreased in the ox-LDL group, showing statistical difference (P <0.05). Compared with the ox-LDL group, the contents of ROS, MDA, and NADPH oxidase decreased, the NO content and the SOD activity increased in the GA 30 mg/L group, the GA 60 mg/L group, and the Vit E group, showing statistical difference (P <0.05). The optimal effects were shown in the GA 60 mg/L group.

CONCLUSIONSGA could obviously inhibit ox-LDL induced synthesis of ROS, lower the contents of MDA, and elevate the levels of NO. Its mechanisms might be associated with increasing the activity of SOD and lowering the activity of NADPH oxidase.

Aorta ; cytology ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Ginkgo biloba ; chemistry ; Humans ; Isoflavones ; pharmacology ; Lipoproteins, LDL ; pharmacology ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Vitamin E ; pharmacology

0.05 for all).Conclusions The allelic frequencies of LPL gene at Pvu Ⅱ locus in Hei Yi Zhuang were different from those in Han,but the genotypie frequencies in Hei Yi Zhuang were not different from those in Han.There was no significant correlation between the polymorphism of LPL gene at Pvu Ⅱ site and the serum lipid levels in two ethnic groups.

A bacterial strain which could grow well on the substrate of PAEs as the sole source of carbon and energy was isolated from contaminated sludge in the river of WeiFang in ShangDong province and it was designated as JDC-3. Based on the morphology,biophysical and biochemical properties as well as molecular characteristics,this isolate was preliminarily identified as Delftia sp.. A fragment of phthalate dioxygenase gene was successfully amplified from the genus of Delftia for the first time using a set of degenerate primers. Meanwhile,the degradation capability of JDC-3 was determined by HPLC using DMP as test substrate. The results showed that the optimal pH and temperature were at 7.0～8.0 and 30?C～35?C respectively. The degradation kinetics of JDC-3 was studied in different initial DMP concentration under optimal conditions. The results indicated that the degradation dynamic equation was ln C =-0.06837 t + A when DMP concentration was lower than 300 mg/L,with half life of 12.48 h. The degradation rate decreased and half life of JDC-3 prolonged as the initial concentration kept on increasing.

Through the analysis on the methods of medicinal paste preparation, the irritation of skin to medicine and the plaster materials adopted in acupoint plaster therapy for the prevention of winter-attacked disease in summer, the acupoint plaster materials and devices were improved. According to the differences in age, illness condition, acupoint and medicinal irritation of patients, the high-dosage, moderate-dosage and low-dosage series of medicine were prepared in proportion; 2. 5 mL and 5 mL syringes were manually reconstructed as the pushers for the delivery of the medicine paste of different specifications. The new-type materials such as spun-bonded non-woven fabrics, transparent dressing film and spun-laced non-woven skin-color stick plaster were adopted. In the operation, the medicine was classified and prepared more specifically. The dedicated acupoint plaster was characterized as less in skin irritation, breathable in property, convenient in practice and proper in stickiness. The plastic anti-seepage film in the middle and the medicine storage pool for stabilizing medicinal paste could avoid drug leakage. The medicinal paste pusher could achieve the even size, proper thickness and precise dosage of the paste. The new-type plaster material and the self-prepared innovated plaster device contribute to the development of acupoint plaster therapy in clinical application.
Acupuncture Points ; Dosage Forms ; Drug Therapy ; instrumentation ; methods ; Drugs, Chinese Herbal ; administration & dosage ; Humans

OBJECTIVETo construct a multiple myeloma (MM)-specific APE1siRNA expression vector, and detect the specific knock-down effect of the siRNA on expression of APE1 protein.

METHODSAPE1siRNA cDNA sequence was designed, synthesized and inserted into pSilencer 2.0-U6 linear expression vector. pSilencer APE1siRNA was digested by enzyme EcoRI and BamHI, then linear vector and IgP fragments were conjugated by T4 DNA ligase. pSilencer IgP-APE1siRNA and pSilencer IE-IgP-APE1siRNA were digested by enzyme EcoRI or XhoI. Linear vector and IE or Kappa fragments were conjugated by T4 DNA ligase. Then a MM specific pSilencer K-IE-IgP-APE1siRNA was cloned. The recombinant products were identified by DNA sequencing and enzyme digestions at each step. pSilencer K-IE-IgP-APE1siRNA plasmid was transfected to KM3, HOS, MDA-231 cells by liposome. APE1 gene silence induced by RNAi was analysed by Western blot.

RESULTSAPE1 protein in KM3 cells could be knocked down effectively and specifically by pSilencer K-IE-IgP-APE1siRNA vector. After 2 days, the level of APE1 protein in KM3 cells transfected with siRNA was 0.118 +/- 0.047, while that transfected with plasmid only was 0.988 +/- 0.029. The efficiency of gene silence was 90%.

CONCLUSIONA MM specific APE1siRNA expression vector was successfully constructed.

Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Genetic Vectors ; genetics ; Humans ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection