Magnetic activated cell sorting (MACS) is considered as a flexible, fast, specific and simple cell sorting system that can separate target cells effectively according to specific markers on the cell surface, for which it has won a wide clinical application. MACS offers a new platform for male infertility research, as well as a novel idea for applying this technology in the optimization of semen quality and the isolation of germ cells. This article briefly introduces the basic principles of MACS, and summaries its present and potential clinical application in male infertility research, as in spermatozoa selection and cryopreservation, and the isolation of spermatogonial stem cells and germ cells.
Cell Separation ; methods ; Flow Cytometry ; Humans ; Infertility, Male ; Magnetics ; Male ; Semen Analysis

OBJECTIVETo study osteoblastic phenotype expression of New Zealand rabbit skin fibroblasts transfected with mouse core binding factor a1/osteoblast specific transplanting factor-2 gene (Cbfa1/Osf2).

METHODSCbfa1/Osf2 gene, engineered into eukaryotic expression vector pSG5, was introduced into New Zealand rabbit skin fibroblasts with catholyte liposomes-Lipofectamine 2000. Meanwhile, those transfected pSG5 and un-transfected were set the control groups. The expression of Cbfa1 gene, osteocalcin (OCN) gene, alkaline phosphatase (ALP) gene and pre-peptide 2 alpha gene of collagen type I were detected by RT-PCR assay. Cbfa1 protein was detected by Western-Blot assay, in-cell ALP activity by p-nitrophenyl phosphate (PNPP) assay and OCN content in the supernatant by radio-immunity method. The ossification nodules was detected by Alizarin-Red staining and scanning electron microscope.

RESULTSCbfa 1mRNA and Cbfa1 protein were expressed in New Zealand rabbit skin fibroblasts transfected with pSG5-Cbfa1/Osf2 from the first day to the fifth day, but they were not detected in the control groups. In the pSG5-Cbfa1/Osf2 transfected group, the expression of ALP gene and OCN gene were respectively induced from the third day and the forth day, pre-peptide 2 alpha gene of collagen type I was enhanced from the third day. From the sixth day, ALP activity greatly increased, OCN strongly secreted, and they were maintained at a high level for about 4 weeks, and the difference was significant compared with the control group (P < 0.05). On the forty-second day, ossification nodules were found on the surface of pSG5-Cbfa1/Osf2 gene transfected cells.

CONCLUSIONSNew Zealand rabbit skin fibroblasts transfected with pSG5-Cbfa1/Osf2 can express osteogenesis-related genes and proteins, and form ossification nodules on their surface.

Alkaline Phosphatase ; biosynthesis ; genetics ; Animals ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; biosynthesis ; genetics ; Fibroblasts ; cytology ; physiology ; Gene Expression ; Genetic Vectors ; Mice ; Osteocalcin ; biosynthesis ; genetics ; Osteogenesis ; genetics ; physiology ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate the influence of high glucose on Porphyromonasgingivalis (Pg) lipopolysaccharide (LPS) stimulating human gingival fibroblasts (HGF) to secret the cytokines.

METHODSHGF were obtained from the primary culture of the tissue explants. Cells were divided into four groups, low glucose (5.5 mmol/L) + 1 mg/L Pg LPS (group A);low glucose + 10 mg/L Pg LPS (group B); high glucose (25 mmol/L) +1 mg/L Pg LPS(group C); high glucose+10 mg/L Pg LPS (group D). The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in cell supernatants were detected by enzyme- linked immunosorbent assay at 6 h and 12 h. The expressions of toll-like receptor 2, 4 (TLR-2, 4) were examined by real-time polymerase chain reaction. After pretreatment with anti-TLR2 and anti-TLR4 monoclonal antibody in HGF, TNF-α and L-1β levels were detected.

RESULTSTNF-α concentration increased obviously in high glucose 6 h and 12 h after Pg LPS stimulation (P < 0.01). IL-1β secretion increased (P < 0.01). Meanwhile, TLR2, 4 mRNA expression increased, especially in high glucose+10 mg/L Pg LPS (P < 0.01). After inhibition of the TLR2, 4 in high glucose + 10 mg/L Pg LPS respectively, TNF-α level [(297.16±11.49), (390.01±12.81) ng/L] decreased (F = 166.02, P < 0.01), and IL-1β level [(49.90±4.08), (99.35±5.01) ng/L] also decreased (F = 153.51, P < 0.01).

CONCLUSIONSHigh glucose may promote Pg LPS to stimulate the secretion of TNF-α and IL-1β through regulating TLR2, 4 expression, which suggests that the elevating blood glucose precipitate in aggravating the process of periodontal disease.

Antibodies, Monoclonal ; Drug Synergism ; Fibroblasts ; drug effects ; metabolism ; Glucose ; administration & dosage ; pharmacology ; Humans ; Interleukin-1beta ; metabolism ; Periodontal Diseases ; etiology ; Polysaccharides, Bacterial ; toxicity ; Porphyromonas gingivalis ; chemistry ; Time Factors ; Toll-Like Receptor 2 ; immunology ; metabolism ; Toll-Like Receptor 4 ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

To explore the causes and solutions of common faults in the operation of the of Yi Kang A1 series lower limb intelligent feedback training system,and to conduct a comparative study on the structural principle and fault maintenance of the two models.Through the analysis of equipment spasm error,USB disconnection error,inability to access the game,black screen of display,splash screen or no display,standing numerical deviation and proprioceptive numerical deviation and other failure cases,gradually investigate software setting,hardware inspection,sensitivity setting,USB connection testing,game file testing,VGA interface testing,angle sensor testing and pressure sensor testing,etc.,to handle and solve the faults in a timely manner to ensure the equipment in good condition,to improve the normal use rate of equipment and ensure the timely rehabilitation of patients.

Objective To investigate the perinatal complications, birth defects and growth of children conceived through intracytoplasmic sperm injection (ICSI). Methods A total of 575 children conceived by ICSI in our reproductive medical center, were studied. The follow-up study would include items as pregnant complications, neonatal complications, birth defects in perinatal period, subsequently detected birth defects, body weight and body length/height growth. Results Prematurity and low birth weight of ICSI children were higher in the multiple births than in the singleton births. The rates of materal gestational hypertension, neonatal asphyxia, respiratory distress syndrome, infection diseases were higher in the multiple pregnancies than in the singleton pregnancies(P＜0.05). Eleven ICSI children had died. Ten of them died in the neonatal period and they were preterm infants. One fullterm singleton ICSI child died of hepatoblastoma at the age of 2. The rate of birth defects in perinatal period was higher in ICSI children of multiple pregnancies than in the general population (P＜0.05). The body weight and body length/height of most ICSI children had obtained the standard range between 1 to 3 year-olds. Conclusion The higher rates of perinatal complications in ICSI children were closely related to multiple pregnancies.

Objective To explore the role of infection control nurses in healthcare-associated infection(HAI)man-agement,and provide basis for HAI management.Methods Through setting up infection control nurses in clinical departments of the whole hospital,clarifying their responsibilities and duties,training,and supervising them,the effect of infection control nurses on HAI management was observed.Results A total of 67 infection control nurses were set up in the clinical departments of the whole hospital,HAI management knowledge among health care work-ers (HCWs)in 26 departments improved significantly,scores of HAI management knowledge among HCWs in April and December was compared,difference was statistically significant (Z = - 2.193,unilateral P = 0.014). Hand hygiene compliance rate of HCWs improved from 83.35%(1817/2180)in April to 89.53% (2002/2236)in December,difference was statistically significant(χ2 =36.13,P <0.01).A total of 56670 hospitalized patients were monitored from April to December 2015,the total length of hospital stay was 411164 days,utilization rate of three catheters was 27.18%,three catheter-related infection rate was 0.74‰.The median scores of supervision on HAI management in clinical departments improved from 95.30 in May to 97.70 in September(P <0.05).Conclusion Setting up infection control nurses is of great significance to strengthen the HAI management organization and pro-mote the quality of HAI management.

OBJECTIVETo study the distribution and drug resistance of methicillin resistant Staphylococcus strains in various specimens of inpatients in burn wards, and to provide reference for clinical treatment.

METHODSBacteria were isolated from specimens of wound exudate, blood, sputum, and bronchoalveolar lavage fluid etc., which were collected from patients hospitalized in our burn wards from January 2008 to December 2010. The bacteria were routinely cultured and identified. Drug resistance of the Staphylococci to 15 antibiotics commonly used in clinic was identified by K-B disk diffusion method. Data were processed with statistical software WHONET 5.5. The homology of 40 strains of methicillin resistant Staphylococcus aureus (MRSA) was analyzed by pulsed-field gel electrophoresis (PFGE).

RESULTSAltogether 386 strains of Staphylococcus were isolated, including 196 strains of Staphylococcus aureus and 190 strains of coagulase negative Staphylococcus. The mean annual isolation rates of MRSA and methicillin resistant coagulase negative Staphylococcus (MRCoNS) were respectively 73.00% (143/196) and 74.20% (141/190). The resistance rates of MRSA and MRCoNS to β-lactams drugs, such as penicillin, oxacillin, cefazolin, and cefuroxime were 100.00% in every year. No Staphylococcus strains resistant to vancomycin, teicoplanin, or linezolid were found. Three different PFGE patterns A, B, and C were identified among 40 MRSA strains, including 33 strains of type A (30 strains in sub-type A1 and 3 strains in sub-type A2), 6 strains of type B (respectively 3 strains in sub-types B1 and B2), and 1 strain of type C.

CONCLUSIONSThe isolation rates of MRSA and MRCoNS were high in our burn wards from January 2008 to December 2010. All of them showed strong drug resistance property, and they were multidrug resistant. The most prevalent strain was PFGE type A.

Burns ; microbiology ; Drug Resistance, Multiple, Bacterial ; Humans ; Methicillin-Resistant Staphylococcus aureus ; drug effects

OBJECTIVEThe present study was to evaluate the audiological characteristics of infants with normal auditory brainstem response thresholds in click and abnormal transiently evoked otoacoustic emissions. Relationships between test results of distortion product otoacoustic emissions (DPOAE) and other hearing testing methods were also evaluated.

METHODSThe participants consisted of eighty-nine infants, with a total of 123 ears. All participants' TEOAE screening results were abnormal but diagnostic click ABR results were normal. The participants were classified into the following groups based on the test results from distortion product otoacoustic emissions: group A (normal all-frequency), group B (abnormal low-frequency), group C (abnormal high-frequency), and group D (abnormal all-frequency).

RESULTSObtained from these groups were compared to results of other hearing tests including the latency of ABR wave I, 40 Hz auditory event related potential (40 Hz AERP), 226 Hz and 1000 Hz tympanometry, and acoustic reflex. Results In six hearing tests in the 123 ears, seven ears (5.7%) were normal, while 116 ears (94.3%) were abnormal. No significantly differences were detected between boys (93.9%) and girls (95.1%), as well as between left (93.1%) and right ears (95.4%). The proportion of abnormal test results ranked as follows: 59 ears in group D (48.0%), 34 ears in group B (27.6%), 20 ears in group A (16.3%), and 10 ears in group C (8.2%). The highest abnormal rates in groups A, B and D were acoustic reflex, which were 40.0% for group A, 55.9% for group B and 66.1% for group D respectively. The highest abnormal rate in group C was the latency of ABR wave I (50.0%). Distribution of low-frequency hearing loss in each group was mainly mild. However, one ear in group B was moderate hearing loss, six ears in group D were moderate hearing loss, and one ear in group D was severe hearing loss.

CONCLUSIONSThe present study showed that, of which infants with normal thresholds of ABR failed the hearing screening, comprehensively audiology assessment is needed. And of which infants with normal DPOAE in full frequency or abnormal in high frequency region or low frequency region need to be followed up.

Auditory Threshold ; Evoked Potentials, Auditory, Brain Stem ; Female ; Hearing Tests ; methods ; Humans ; Infant ; Infant, Newborn ; Male ; Neonatal Screening ; methods ; Otoacoustic Emissions, Spontaneous ; Reflex, Acoustic

Objective To investigate the association between cutaneous adverse drug reactions (CADRs) caused by antiepileptic drugs and HLA-B*1502 allele. Methods In 31 epileptic patients presented to the Epilepsy Clinic of the Second Affiliated Hospital of Guangzhou Medical College between January 2007 and May 2008, 13 had CADR to carbanazepine (CBZ) including 6 with Stevens-Johnson syndrome (SJS) and 7 with mild maculopapular exanthona (MPE);15 were CBZ-tolerant, and 3 had lamotrigine (LTG)-indueed MPE. All the patients underwent examinations using polymerase chain reaction with sequence specific palmers to analyze HLA -B*1502 allele frequencies, with 30 healthy subjects without a history of using CBZ or LTG as the control. Results HLA-B*IS02 allele frequency was 100% (6/6) in patients with CBZ-SJS, 57% (4/7) in patients with CBZ-induced MPE, and 33% (1/3) in patients with LTG-induced MPE. The frequency was 7% (1/15) in CBZ-tolerant patients and 10% (3/30) in the control subjects. Compared with the CBZ-tolerant patients and the control subjects, the patients with CBZ-induced SJS and MPE had significantly increased HLA -B*1502 allele frequency (P<0.05). Conclusions HLA-B*1502 allele is associated with CADRs to CBZ in epileptic patients.