Action Potentials ; drug effects ; Animals ; Cadmium ; toxicity ; Guinea Pigs ; Male ; Myocytes, Cardiac ; drug effects ; physiology

Objective To explore the temporal and spatial expression pattern of tenascin-c(tnc) and tenascin-w(tnw) in zebrafish early development,to further explore the role of tenacsin in zebrafish embryo development,and the association between them.Methods Zebrafish embryos at 2 hours post fertilization(hpf),4 hpt,8 hpt,10 hpf,24 hpt,48 hpr,72 hpf and 7 days post fertilization(dpf) were collected to extract RNA for reverse transcription-polymerase chain reaction(RT-PCR) and fix the embryos at different stages for in situ hybridization.Temporal and spatial expression pattern of tnc and tnw on different stages of zebrafish early development was observed.Results tnc and tnw all expressed in zebrafish from 24 hpf to 7 dpf,but did not expressed from 2 hpf to 10 hpf.Tnc expressed at pharyngeal arch,notochord,somite in 24 hpf,then weakly expressed at somite,but highly expressed at otic vesicle,pectoral fin and hindbrain in 48 hpf,and tnc was expressed at hindbrain,pharyngeal and notochord and disappeared at somite and pectoral.tnw expressed at hindbrain,midbrain and otic vesicle in 24 hpf,expressed at somite,notochord,hindbrain,otic vesicle and pharyngeal in 48 hpf.In 72 hpf,tnw expressed weakly at somite and notochord.Conclusions Zebrafish tnc and tnw have special temporal and spatial expression pattern,and share partial overlapping expression pattern.

Objective To explore the expression pattern of tenascin-c(tnc)gene in zebrafish embryo abnormal development which was induced by ethanol,and to further understand the function of tnc gene in embryo develepment.Methods Zebrafish were treated with ethanol at different concentration from 100 to 500 mmol/L,and embryos at 24 and 48 hours were collected and fixed,then tnc expression pattern was observed by in situ hybridization and reverse transcriptase-polymerase chain reaction(RT-PCR).Results The result of RT-PCR showed that ethanol at 100 and 200 mmol/L could increase the expression of tnc,while the result of in situ hybridization showed that,while ethanol at 300 mmol/L and above decrease the expression of tnc in presumptive position at 24 hours,and ethanol at 100 mmol/L and above caused increase expression of tnc in zebrafish heart.Conclusions tnc is increased when treated with 100 and 200 mmol/L ethanol and is presented in the abnormal development of hearts of zebrafish,which can promote the normal development of embryos in some degrees.The expression pattern of tnc in pathologic state is highly conserved in all vertebrate,and in adult and embryos as well.

BACKGROUND: The quality of chest compressions can be significantly improved after training of rescuers according to the latest national guidelines of China. However, rescuers may be unable to maintain adequate compression or ventilation throughout a response of average emergency medical services because of increased rescuer fatigue. In the present study, we evaluated the performance of cardiopulmonary resuscitation (CPR) in training of military medical university students during a prolonged basic life support (BLS). METHODS: A 3-hour BLS training was given to 120 military medical university students. Six months after the training, 115 students performed single rescuer BLS on a manikin for 8 minutes. The qualities of chest compressions as well as ventilations were assessed. RESULTS: The average compression depth and rate were 53.7±5.3 mm and 135.1±15.7 compressions per minute respectively. The proportion of chest compressions with appropriate depth was 71.7％±28.4％. The average ventilation volume was 847.2±260.4 mL and the proportion of students with adequate ventilation was 63.5％. Compared with male students, significantly lower compression depth (46.7±4.8 vs. 54.6±4.8 mm,P<0.001) and adequate compression rate (35.5％±26.5％ vs. 76.1％±25.1％,P<0.001) were observed in female students. CONCLUSIONS: CPR was found to be related to gender, body weight, and body mass index of students in this study. The quality of chest compressions was well maintained in male students during 8 minutes of conventional CPR but declined rapidly in female students after 2 minutes according to the latest national guidelines. Physical fitness and rescuer fatigue did not affect the quality of ventilation.

OBJECTIVETo investigate the diagnostic application of molecular detection of enterovirus type 71 (EV71) infection using post-mortem paraffin-embedded tissue.

METHODSTwo autopsy cases of EV71 infection were studied by histopathological and immunohistochemical methods. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the viral RNA in paraffin-embedded tissue samples.

RESULTSCharacteristic features of acute encephalitis were seen in the brain, with most prominent lesions found in the brain stem in both cases. Inflammatory cells were largely CD68-positive microglia with a few CD15-positive neutrophils in the areas of neuronal necrosis. The 5'-untranslated region of EV71 was detected in the medulla by RT-PCR using paraffin-embedded tissues of both cases. Sequencing analysis of the RT-PCR products showed 100% homology to the EV71 strain, recently submitted to the GenBank database from Fuyang, Anhui province.

CONCLUSIONSMolecular detection of EV71 can be performed on formalin-fixed, paraffin-embedded tissue samples from fatally infected patients. Timely and accurate diagnosis of the infection by such molecular approach is crucial for the proper clinical and public health intervention.

5' Untranslated Regions ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Autopsy ; Brain ; metabolism ; Encephalitis ; metabolism ; virology ; Enterovirus A, Human ; genetics ; isolation & purification ; Enterovirus Infections ; metabolism ; pathology ; virology ; Female ; Humans ; Infant ; Lewis X Antigen ; metabolism ; Male ; Paraffin Embedding ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, RNA

OBJECTIVETo study the BRCA1 mutations in patients with early-onset breast cancer and their affected relatives in Guangdong province and explore the relationship between BRCA1 mutation and the expressions of estrogen receptor(ER), progesterone receptor(PR), HER2 and ALN.

METHODSFrom 58 patients with early-onset breast cancer and their affected relatives, the genomic DNA was extracted from the peripheral blood mononuclear cells and the coding regions of the BRCA1 gene was amplified using polymerase chain reaction. BRCA1 gene mutations were screened by denaturing high performance liquid chromatography (DHPLC) and subsequent direct DNA sequencing. The expression of ER, PR, HER2 and ALN were detected with immunohistochemistry and their relations with the gene mutation were analyzed.

RESULTSDisease-related BRCA1 mutations were detected in 2 of the 58 patients, who were younger than 35 years old, including 1 with a novel splice-site mutation (IVS5-1 G-->A). No association was found between this novel mutation and the expressions of ER, PR, HER2 and ALN.

CONCLUSIONThe incidence of BRCA1 mutation is significantly lower in patients with early-onset breast cancer and their affected relatives in Guangdong province than in the Western populations. The novel mutation identified in BRCA1 gene may represent a mutation characteristic of the patients in Guangdong province. BRCA1 gene mutations may not have any relation with the expression of ER, PR, HER2 and ALN.

Adult ; Age of Onset ; Base Sequence ; Breast Neoplasms ; genetics ; China ; DNA Mutational Analysis ; Female ; Genes, BRCA1 ; Genotype ; Humans ; Molecular Sequence Data ; Mutation ; Receptor, ErbB-2 ; genetics ; Receptors, Estrogen ; genetics ; Receptors, Progesterone ; genetics

OBJECTIVETo summarize clinical application of one-stage toenail lengthening in free second toe transfer for reconstruction of the thumb (finger).

METHODSNine patients (male 7, female 2) underwent thumb (finger) reconstruction with second toe transfer were treated by one-stage toenail lengthening technique. Eight were the thumb and 1 was the index finger. Patients aged from 18 to 46 years,with an average of 25 years. A rectangle skin was resected at 0.5 cm away from the eponychium, which was 0.2 cm high and as wide as the toenail. Then stripped U shape flap gently towards proximal end and sutured it. During the operation, the injury of the subcutaneous vascular network should be avoided.

RESULTSSuperficial infection at donor area happened in 1 case and was healed by changing dressings. All the reconstruction thumbs (fingers) had survived completely. 2 to 3 mm extending of toenail length was obtained and the appearance of thumb (finger) was improved. There was no growth deformation of toenail. After 7 to 24 months follow up (the average time 13 months), the appearance of the nail was good.

CONCLUSIONSOne-stage toenail lengthening in free second toe transfer for reconstruction of the thumb (finger), which can obtain a satisfactory appearance of the nail and have no influence on the motion of the reconstruction thumb (finger), is a simple and an effective operative procedure.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nails ; transplantation ; Reconstructive Surgical Procedures ; methods ; Thumb ; surgery

OBJECTIVE: To establish immortalized embryonic fibroblast lines in heat shock transcription factor 1 (HSF1) HSF1-/- and HSF1+/+ mice and to provide experimental models to study the function of HSF1. METHODS: A mammalian expression vector (pSV3neo) containing the SV40 large T antigen was used to transfect the HSF1-/- and HSF1+/+ mouse embryonic fibroblast using Lipofectamine 2000. Colonies were screened by G418 and expanded to immortalized cell lines. PCR was used to detect the integration of the large T antigen with genome in the mouse embryonic fibroblast. Expression of SV40 large T antigen gene in expanded cells was identified by RT-PCR. HSP70 expression was examined by Western blot in the embryonic fibroblast lines. RESULTS: The stable growth and serial propagation were observed in the HSF1-/- and HSF1+/+ cell lines for six months. The mRNA of SV40 T antigen gene expressed in the two cell lines. HSP70 expression could not be induced in the heat-treated HSF1-/- mouse embryo fibroblasts. CONCLUSION The immortalized cells of HSF1+/+ and HSF1-/- mouse embryo fibroblasts are successfully established.
Animals ; Antigens, Polyomavirus Transforming ; pharmacology ; Cell Line ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; Female ; Fibroblasts ; cytology ; Heat Shock Transcription Factors ; Male ; Mice ; Mice, Knockout ; Transcription Factors ; genetics

ObjectiveTo observe the effects of Qingzao Jiufeitang on the expression of adenosine 5′-monophosphate （AMP）-activated protein kinase （AMPK）， mammalian target of rapamycin （mTOR）， and UNC-51-like kinase 1 （ULK1） in lung cancer cells after the application of AMPK inhibitor （compound C）. MethodMale C57BL/6J mice were randomly divided into a model group， a cyclophosphamide （CTX） group （50 mg·kg^-1）， a Qingzao Jiufeitang group （11 g·kg^-1）， an AMPK inhibitor group （10 mg·kg^-1）， and a Qingzao Jiufeitang combined with AMPK inhibitor group （combination group） （11 g·kg^-1+10 mg·kg^-1）. Lewis lung cancer cells were subcutaneously injected into the right axilla to induce a tumor-bearing model. 24 hours after modeling， the mice in the CTX group were intraperitoneally injected once every other day for seven times in total. The mice in the AMPK inhibitor group and the combination group received intraperitoneal injection of compound C， once a day for 14 days. The mice in the Qingzao Jiufeitang group and the combination group were administered orally at the set dose for 14 days before and after modeling. At the end of the experiment， the mice in each group were sacrificed. The tumor-bearing tissues were collected， and the tumor weight of each group was counted. Transmission electron microscopy （TEM） was used to observe the formation of autolysosomes in lung cancer tissues of each group. Western blot was used to detect the protein expression of AMPK， phosphorylated AMPK （p-AMPK）， mTOR， phosphorylated mTOR （p-mTOR）， ULK1， phosphorylated ULK1 （p-ULK1）， microtubule-associated protein 1 light chain 3B （LC3B）， and p62. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of lung cancer in each group. ResultCompared with the model group， the Qingzao Jiufeitang group showed decreased tumor weight （P<0.01）， the formation of autolysosomes under the electron microscope， increased protein expression of p-AMPK， p-ULK1， LC3B， LC3B-Ⅱ， and p-AMPK/AMPK， p-ULK1/ULK1， and LC3B-Ⅱ/LC3B-Ⅰratios （P<0.01， P<0.05）， and reduced protein expression of p-mTOR， p62， and p-mTOR/mTOR ratio （P<0.05）. Compared with the Qingzao Jiufeitang group， the combination group showed no autolysosomes formation under the electron microscope， decreased protein expression of p-AMPK， p-ULK1， LC3B， LC3B-Ⅱ， and p-AMPK/AMPK， p-ULK1/ULK1， LC3B-Ⅱ/LC3B-Ⅰ ratios （P<0.05， P<0.01）， and increased p62 protein expression （P<0.05）. HE staining results showed that the pathological changes of lung cancer tissues in the groups with drug intervention were improved compared with those in the model group. ConclusionQingzao Jiufeitang can promote the elevation of LC3B-Ⅱ and decrease the expression of p62 protein， thus inducing autophagy. The mechanism of autophagy initiation may be achieved by the AMPK/ULK1 pathway instead of the mediation by the AMPK/mTOR/ULK1 pathway.

Objective To determine the adherence to and its determinants of methadone maintenance treatment (MMT) among heroin addicts in Dehong prefecture in Yunnan province from 2005 to 2009. Methods A dynamic cohort analysis was conducted with the time of treatment initiation as the time of cohort entry, to calculate the proportion of adhering to the treatment at different time points after initiation of the treatment, and to study the determinants of adherence.Results A total of 3758 had been treated. Among them, 95.8% were males, 75.8% aged between 20-39 years, 90.4% were peasants or unemployed, 57.0% were ethnic minorities, 35.0% were single and 55.5% were married with spouses, 43.9% were illiterate or educated at most primary school. The age of first using drugs averaged at 23.93 years. About 96.3% of the study subjects had used heroin and 21.7% were HIV-infected. The minimum time under MMT was less than 1 month and the maximum 61 months. The median methadone dose at first delivery was 25 ml, with the minimum 1 ml and the maximum 330 ml. By the end of the study or observation period, a total of 1798 patients had withdrawn from treatment and 1960 were still under treatment. The proportions of adherence to or still being under the treatment after 1,3,6,9, 12,24, 36,48 and 60 months treatment were 0.919,0.847,0.756, 0.690, 0.637, 0.519, 0.417, 0.360 and 0.321, respectively. Multiple regression analysis using Cox proportional hazard model indicated that withdraw from the methadone maintenance treatment was significantly associated with location of the treatment clinics, year of treatment initiation, marital status, HIV infection status, methadone dose of first delivery and the result of last urine test for heroin use. Conclusion MMT attendants in Dehong prefecture had a relatively high withdraw rate and low adherence rate. More efforts are needed to provide tailored counseling and education to MMT attendants, to provide family and community support, appropriate methadone dose at first delivery, and to better coordinate with local police department.