A human diploid fibroblast cell line derived from normal abdominal skin of a 17-year-old female patient with ovary carcinoma was established and designated as HSF 79-2. The cultivation of skin fragments was employed in the primary explantation. The medium used was McCoy 5A supplemented with 15～25% calf serum. The grown fibroblasts were succesively subcultured at 1 to 2 split and stopped at the 45th to 54th passage.Having reached confluence the cells still had the ability to divide during 16 months observation if the medium was changed periodically. They displayed the property of dense growth and forming heavy, multilayer sheets, which became a visible membrane to the naked eyes. Under histological examination the membrane had the appearance of connective tissue. Part of the cells were subcultured again after 3,6, and 12 months maintenance in the same culture flask. Their growth character, ploidy, and the generation time were similar to that of the cells passaged normally.The characteristics of this cell line mentioned above appear to be shared by other diploid fibroblasts. It might be used for preserving cell lines and as a model for studying cell motility and differentiation.

Objective To explore the effects of L-carnitine on newborn with hypoxic ischemic myocardial injury.Methods Sixty cases with hypoxic ischemic myocardial injury were divided into two groups,including control group (n =30) receiving fructose sodium diphosphate injection,and experimental group (n =30) receiving both fructose sodium diphosphate and L-carnitine injection for 7 days.The clinical symptoms,changes of heart rates,blood pressure,volume of urine,lactic acid dehydrogenase,creatine kinase,creatine kinase-MB,cardiac troponin Ⅰ and electrocardiogram were investigated before and after treatment.Results Treatment with L-carnitine improved the symptoms of hypoxic ischemic myocardial injury,the total effective rate was 93.3％ (28/30),versus 66.7％ (20/30) in control group,the difference was statistically significant(x2 =6.667,P =0.01).Heart rates of the experimental group were significantly improved after treatment(t =-6.131,P ＜0.01),compared with the control group,there were no significant differences (P ＞ 0.05).Blood pressure and urine volume of experimental group both increased after treatment (P ＜0.05),compared with the control group,the differences were statistically significant (P ＜ 0.05).Creatine kinase and creatine kinase-MB significantly decreased after treatment,compared with the control group (P ＜ 0.05).A downward trend of cardiac troponin Ⅰ was observed in both groups,however,there was no statistically significant difference before and after treatment (P ＞ 0.05).After treatment,electrocardiogram improvement rates were no significantly different between the two groups (P ＞ 0.05).Conclusion L-camitine can contribute to protect hypoxic ischemic myocardial from further injury.

Objective To evaluate the role of μ opioid receptor exon 7 in the analgesic efficacy of endomorphin-2 in rats.Methods Twenty-four male Sprague-Dawley rats in which IT catheters were successfully implanted,weighing 220-260 g,were randomly divided into 3 groups (n =8 each) using a random number table:normal saline control group (group C),negative siRNA control group (group N-siRNA) andμ opioid receptor exon 7 siRNA group (group E7-siRNA).In C,N-siRNA and E7-siRNA groups,30μl saline solution,negative siRNA plasmid 20 μl + lipofectamine 2000 (10 μl),and μ opioid receptor siRNA plasmid 20μ1 + lipofectamine 2000 (10 μl) were intrathecally injected once a day for 3 consecutive days.The mechanical pain threshold was measured on 4th day (baseline).Endomorphin-2 10 μg was injected intrathecally at 1 h after measurement of the pain threshold.The mechanical pain threshold was measured at 5,20,40 and 60 min after endomorphin-2 injection,and the analgesic efficacy was calculated.Results There was no significant difference in the baseline pain threshold among the three groups.Compared with group C,no significant difference was found in the analgesic efficacy at each time point after endomorphin-2 injection in group N-siRNA,and the analgesic efficacy was significantly decreased at 5 and 20 min after endomorphin-2 injection in group E7-siRNA.Conclusion μ opioid receptor exon 7 is involved in the analgesic efficacy of endomorphin-2 in rats.

Objective To evaluate the effect of dexmnedetomidine pretreatment on apoptosis in hippocampal neurons of rats undergoing one-lung ventilation (OLV).Methods Ninety adult male SpragueDawley rats,aged 10-11 months,weighing 260-300 g,were divided into 3 groups (n=30 each) using a random number table:two-lung ventilation (TLV) group,group OLV and dexmedetomidine pretreatment group (group D).Dexmedetomidine 25 μg/kg was injected intraperitoneally at 45 min before OLV in group D.After tracheal intubation,the animals were ventilated in volume-controlled mode.OLV was performed for 90 mnin followed by 30 min of TLV in OLV and D groups.TLV was performed for 120 min in group TLV.On 1,3 and 7 days after ventilation,6 rats in each group were selected,and Morris water maze test was carried out to evaluate the cognitive function.The swimming speed,time of staying at the target quadrant,and frequency of crossing the platform quadrant were recorded.Six rats in each group were selected immediately after ventilation and sacrificed,the hippocampi were removed for detection of cell apoptosis,and the apoptosis index was calculated.Immediately after ventilation and on 1,3 and 7 days after ventilation,6 rats in each group were selected and sacrificed,and the hippocampi were removed for determination of phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2),phosphorylated cyclic adenosine monophosphate response element-binding protein (pCREB),Bcl-2 and Bax expression.The ratio of Bcl-2 expression to Bax expression (Bcl-2/Bax ratio) was calculated.Results Compared with group TLV,the time of staying at the target quadrant was significantly shortened,the frequency of crossing the platform quadrant was decreased,the apoptosis index was increased,the expression of pERK1,pERK2,pCREB and Bcl-2 was down-regulated,the expression of Bax was up-regulated,and Bcl-2/Bax ratio was decreased in group OLV (P＜0.05 or 0.01).Compared with group OLV,the time of staying at the target quadrant was significantly prolonged,the frequency of crossing the platform quadrant was increased,the apoptosis index was decreased,the expression of pERK1,pERK2,pCREB and Bcl-2 was up-regulated,the expression of Bax was down-regulated,and Bcl-2/Bax ratio was increased in group D (P＜0.05 or 0.01).Conclusion Dexmedetomidine pretreatment decreases apoptosis in hippocampal neurons through activating ERK/CREB signaling pathway,thus reducing cognitive dysfunction of rats undergoing OLV.

Objective To assess the efficacy of oral erythromycin on the functional gastrointestinal dysmotility in neonates.Methods In this double-blind,randomized,placebo controlled trial,90 neonates consecutively admitted to the neonatal intensive care unit of Shenzhen Children's Hospital from Jan 2009 to Dec 2011 were enrolled and randomly divided into low-dosage erythromycin group ( LE group,n =30 ),highdosage erythromycin group ( HE group,n =30 ) and control group ( n =30).Patients received either erythromycin ( 3 mg/kg or 10 mg/kg) or equivalent normal saline with oral or nasal feeding every 8 hours one time for 14 d.The time to achieve half,three-quarters,and full enteral nutrition,the time of parenteral nutrition,and hospital length of stay were compared among each group.Results The time to achieve half,three-quarters,and full enteral nutrition in HE group [ ( 3.0 ± 0.5 ) d,( 6.2 ± 0.7 ) d,( 8.2 ± 1.0 ) d ] and in LE group [(6.2±0.5) d,(8.3 ±0.6) d,(10.6 ±1.1) d] were shorter than that in control group [(8.1 ±0.4) d,( 13.5 ± 1.0) d,( 15.7 ± 1.2) d] ( P ＜ 0.05 ).The duration of parenteral nutrition [ ( 14.2 ± 1.4) d vs (9.3 ± 1.2) d vs (7.8 ± 1.1 ) d ] and hospital length of stay [ ( 13.0 ± 1.4 ) d vs ( 8.1 ± 0.8 ) d vs ( 6.8 ±0.7) d] were significantly prolonged in control group compared with LE and HE groups,and there were significant differences among the three groups ( P ＜ 0.05).The incidence of liver injury and septicemia during the treatment of erythromycin were similar between HE group and LE group,but it was significantly lower than control group.No serious adverse effect such as prolongation of QT intervals,dysrhythmia associated with erythromycin treatment was found.Conclusion Oral erythromycin can be considered as a treatment for neonates with functional gastrointestinal dysmotility who fail to establish adequate enteral nutrition,and highdosage oral erythromycin is more effective than low-dosage.

Aim To explore the proteomics mechanism of the differentiation induction effect of 4-amino-2-trif-luoromethyl-phenyl retinate(ATPR)on human leukemi-a K562 cells. Methods Human leukemia K562 cells were incubated with the same concentration (1 × 10 - 6 mol·L - 1 ) of ATPR or ATRA for 48 hours. The total cell proteins were collected, purified and digested by trypsin, solid phase extraction, and the peptides were detected by ESI-LC-MS / MS. The difference of the pro-tein expression between the cells treated with ATPR and ATRA was compared by using the Discoverer Pro-teome 1. 2 software, and the molecular function, the biological process and other information of those pro-teins were analyzed based on the DAVID, KEGG, STRING databases. Results 120 specific proteins were identified only in the ATPR group, 143 only in the ATRA group, and 422 other proteins in both groups. Results of DAVID analysis showed that ATPR-induced specific proteins were mainly involved in 39 biological processes of proteins and macromolecules metabolism, protein transport and localization and so on. Results of KEGG analysis revealed that ATPR-in-duced proteins participated in signal pathways, mainly metabolic pathways, PI3K-Akt signal pathway, TGF-beta signal pathway and other pathways in cancer. String protein interaction network analysis displayed that ATPR-induced proteins, like EIF3A, EIF6, RPL3, RPL8, RPL13, RPL7A, RPL21, RPS3, RPS14, NACA, BTF3, NHP2L1, PPP2CA proteins had direct interactions with more than or equal to 10 associated proteins. Conclusion The differentiation induction effect of ATPR on K562 cells might be as-cribed to the ATPR-induced proteins interaction net-work and the specific central proteins it induced, which are involved in the regulation of cell prolifera-tion, differentiation and apoptosis.

Objective To evaluate the method and result of individual deglutition training on dysphagia after Stroke. Methods 36 patients with dysphagia after stroke were randomized into observation group (20 patients receiving individual deglutition training and routine therapy of neurology) and control group (16 patients receiving basal deglutition training and routine therapy of neurology).Results After treatment for 4 weeks, the score of dysphagia in observation group were higher than in control group (P<0.05). Conclusion Individual deglutition training could significantly improve the swallowing function of the patients with dysphagia after stroke.

Objective To indentify Streptobacillus moniliformis isolated from the knee joint pus by 16S rRNA gene sequencing and biochemical reactions and explore the clinical value of the method. Methods The bacterial 16S rRNA gene sequence-based identification, bacterial morphology, VITEK 2 automate systems, API 20NE strips, API 20E strips and API 50CH were performed to identify the rare bacteria. Results The bacteria grew slow on blood agar and chocolate agar and were inhibited on Maconkey agar. The bacterial colony on blood agar tookes the form of 1~2 mmomelette, which was translucent and moist with circular protrusion and smooth edges. They were Gram-staining negative and in catenation, its thalli 1~3μm, round, oval or fusiform. Vitek 2 GN-13, API 20NE and API 20E were unable to reach the identification of the bacteria. 16S rRNA gene sequencing showed the bacteria were similar to streptobacillus moniliformis by 100%. Conclusion The rare bacteria isolated from left knee joint are streptobacillus moniliformis. 16S rRNA gene sequences combined with the biochemical reactions is accurate in the identification of these bacteria.

ScanningElectronMicroscopy/EnergyDispersionSpectrum(SEM/EDS),ElectronProbe Microanalysis ( EPMA ) and Optical Microscopy ( OM ) are the traditional methods for inclusion type identifying at present. The disadvantages such as inconvenient sample preprocessing, low sensitivity and time consuming limited their application. Laser-Induced Breakdown Spectroscopy ( LIBS) is not only performing in bulk analysis field but also in elemental distribution and depth profile information field, so it causes great concern in material science. In this paper, 34CrNiMo6 steel and heavy railway steel samples were analyzed by LIBS in scanning mode to characterize the inclusion type. For 34CrNiMo6 steel, 2D intensity distribution and channel combination revealed the spectra line intensity of Mn and S were abnormal high simultaneously at some region, which indicated there were some MnS inclusion existed in these samples. And for heavy railway steel, 2D distribution and channel combination revealed the spectra line intensity of Si, Ca, Mg and Al were abnormal high simultaneously at some region, showed the existence of Si-Al-Ca-Mg inclusion in these samples. The SEM/EDS analysis result of above-mentioned samples showed agreed well with LIBS.

Objective To observe the effect of scalp acupuncture on cognitive and motor functions of stroke patients in recovery stage.Methods80 stroke patients with cognitive disorder were randomly divided into the scalp acupuncture group (n=41) and control group (n=39). All patients of two groups were treated with routine therapy and rehabilitation training, but the cases of the scalp acupuncture group were added with scalp acupuncture. The cognitive function, motor function and activities of daily living (ADL) of all patients were assessed by delitescence and amplitude of event-related potential (ERP) P300, Fugl-Meyer Assessment (FMA), Barthel Index (BI) respectively before and 3 months after treatment.ResultsAfter treatment, P300 delitescence of the patients in the scalp acupuncture group shorten to 38 ms in average while in the control group shorten only to 17 ms, the therapeutic effect of the scalp acupuncture group was superior to that of the control group ( P<0.05). The scores of FMA and BI of the patients in the scalp acupuncture group were also better than those in the control group ( P<0.05).ConclusionScalp acupuncture can efficiently improve the cognitive function of stroke patients in the recovery stage, and promote the recovery of motor function and ADL.