Objective To study the influencing factors and quality control of chromosome preparation during chromosome aberration test in vitro,and to summarize and analyze the method and key points of successful preparation of chromosome specimens in vitro. Methods Chinese hamster lung cells(CHL)were used for cell culture and chromosome preparation. Mitomycin and cyclophosphamide were used as positive mutagens. After routine hypotonic treatment,fixation, and squash preparation, finally, to read the film under the microscope. Results The CHL chromosome aberration test showed that both the chromosome aberration rates of mitomycin- and cyclophosphamide-treated cells were significantly increased(>20%),while the aberration rates in the negative control group were less than 5%, either with and without metabolic activation. The success rate was high and the prepared chromosomes were well dispersed with a moderate length. Conclusions Many factors can affect the specimen preparation in chromosome aberration test. Every step is very important,and it should be strictly following the operating procedures. It is of importance to grasp the principle of each step and to operate carefully,patiently and scientifically in order to prepare good specimens.

Objective To study the function of ten-eleven translocation 2 (Tet2) in γglobin gene expression in patients with β-thalassemia.Methods Gamma globin expression was induced by 5-azacytidine and Tet2 gene expression was knocked down by short hairpin RNA (shRNA) in a human immortalized myelogenous leukemia K562 cell line.The global 5-hydroxymethylcytosine (5hmC) level was measured by an ELISA kit.5hmC level of γglobin gene was quantified by sulfite sequencing.The mRNA level of Tet2,γglobin,and related transcription factors Nfe4 and Klfl were quantified by real-time PCR.Results Tet2 knockdown resulted in a decreased global 5hmC level from 0.14％ to 0.03％ as of the control group in K562 cells.The expression of γ globin was enhanced after 5-azacytidine treatment in vitro.However,γglobin mRNA level in Tet2 knockdown cells was only 55％ as that in control group.The CG sites on γ globin gene were unmethylated.As Tet2 was down-regulated,the expression levels of Nfe4 and Klf1 decreased by about 80％ and increased to 3.5 folds,respectively.Conclusions Tet2 appears to maintain 5hmC level and facilitates γ globin gene activation.Moreover,Tet2 more likely regulates γglobin expression via affecting transcription factors rather than the gene itself.Thus,Tet2 could be a potential therapeutic target for β thalassemias.

OBJECTIVE To investigate the expression of zonula occluden-1(ZO-1) in hypopharyngeal squamous cell carcinoma(HSCC) and its clinical significance.METHODS Immunohistochemistry with tissue microarrays was used to detect the expression of the ZO-1 protein in 52 cases of HSCC.The correlation of prognosis and clinicopathological features was analyzed.RESULTS The expression of ZO-1 protein in HSCC was lower than that in adjacent tissues,its expression was closely related with tumor differentiation,tumor stage,tumor location and lymphatic metastasis.The Log-rank univariate analysis showed that the prognosis of HSCC was related to tumor differentiation,tumor stage,lymphatic metastasis and tumor location.The patients with lower expression of ZO-1 showed the worse prognosis than patients with higher expression of ZO-1.As shown in the Cox regression analyses,tumor differentiation,lymphatic metastasis,the ZO-1 expression and tumor stage were all independent prognostic factors.CONCLUSION The ZO-1 can serve as an independent prognostic factor of HSCC,and the low expression of ZO-1 is closely related to poor prognosis.

OBJECTIVETo explore the disease-causing mutations in a patient suspected for giant axonal neuropathy(GAN).

METHODSTarget sequence capture sequencing was used to screen potential mutations in genomic DNA extracted from peripheral blood sample of the patient. Sanger sequencing was applied to confirm the detected mutation. The mutation was verified among 400 GAN alleles from 200 healthy individuals by Sanger sequencing. The function of the mutations was predicted by bioinformatics analysis.

RESULTSThe patient was identified as a compound heterozygote carrying two novel pathogenic GAN mutations, i.e., c.778G>T (p.Glu260Ter) and c.277G>A (p.Gly93Arg). Sanger sequencing confirmed that the c.778G>T (p.Glu260Ter) mutation was inherited from his father, while c.277G>A (p.Gly93Arg) was inherited from his mother. The same mutations was not found in the 200 healthy individuals. Bioinformatics analysis predicted that the two mutations probably caused functional abnormality of gigaxonin.

CONCLUSIONTwo novel GAN mutations were detected in a patient with GAN. Both mutations are pathogenic and can cause abnormalities of gigaxonin structure and function, leading to pathogenesis of GAN. The results may also offer valuable information for similar diseases.

Amino Acid Sequence ; Child ; Computational Biology ; Cytoskeletal Proteins ; genetics ; Giant Axonal Neuropathy ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Sequence Analysis, DNA

OBJECTIVE:To study the effects of mifepristone on the expression of inflammatory factors in endometriosis (Ems)model rats. METHODS:SD rats were randomly divided into normal group(normal saline),model group(normal saline), mifepristone low-dose,medium-dose and high-dose groups [0.65,1.30,2.60 mg/(kg·d)],with 10 rats in each group. Except for normal group,Ems model was induced in other groups,and they were given relevant medicine intragastrically for consecutive 4 weeks. The volume of the ectopic focus was compared before and after treatment. The serum contents of TNF-α,IL-6 and IL-8 were detected by ELISA method. The phosphorylation of NF-κB p65 and the expression of COX-2 and PGE2 were detected by West-ern blot. The expression of NF-κB p65 mRNA in ectopic focus was detected by RT-PCR. RESULTS:Compared with normal group,the volume of the ectopic focus,the serum contents of TNF-α,IL-6 and IL-8,the expression of COX-2 and PGE2 protein, the phosphorylation of NF-κB p65 and the expression of NF-κB p65 mRNA in ectopic focus were increased in model group(P<0.01). Compared with model group,the volume of the ectopic focus,the serum contents of TNF-α,IL-6 and IL-8,the expression of COX-2 and PGE2 protein,the phosphorylation of NF-κB p65 and the expression of NF-κB p65 mRNA in ectopic focus were de-creased in mifepristone groups(P<0.01),in dose-dependent manner. CONCLUSIONS:Mifepristone can reduce the volume of the Ems ectopic focus,via blocking NF-κB signaling pathway and reducing the expression of inflammatory factors.

Animals ; Cell Differentiation ; Hepatocyte Growth Factor ; genetics ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mouse Embryonic Stem Cells ; cytology ; metabolism ; Protein Precursors ; genetics ; metabolism ; Serine Endopeptidases ; genetics ; metabolism

Objective:To explore the correlation between resilience and depression in children with chronic disease.Methods:A total of 109 children from Jun 2014 to Dec 2015 in four hospitals in Shenyang city were randomly selected.The family situation, personal resilience and depression was investigated with Personal Information Questionnaire,Chronic Illness Children’s Resilience Scale,and Children’s Depression Inventory.Results:The average level of resilience was (104??77±11??58).The average level of depression rating scale was (10??86±5??92).There was statistical difference in average level of resilience in the depression group and non?depression group (P<0??05).There was negative correlation between depression and the level of resilience,toughness of the children’s personality characteristics, coping style, interpersonal relationships in children with chronic disease (r =-0.418,-0??461,-0.428,-0.345).Conclusion:Nurses should take various measures to improve the resilience of the children,in order to enhance the children’s disease management behavior,so as to promote their mental health.

OBJECTIVE: To study the efficacy of treatment with microsurgery in combination with local injection of type A botulinum toxin for vocal process granuloma. METHOD: 28 patients with vocal process granuloma received endotracheal intubation under general anesthesia. The lesion was removed with micro-scissor and CO2 laster under a self-retaining laryngoscope and microscope. The incision and mucous membrane surrounding the wound was closed with 8-0 absorbable suture. 4-point injection of botulinum toxin type A 8-15 u was then performed along the thyroarytenoid muscle and arytenoid muscle of the same side. Postoperative medication was administered based on disease causes. RESULT: All patients experienced vocal cord dyskinesia of the injected side 2-3 days after surgery. At 1 month after the surgery, wound healing was good in all the 28 patients, and the vocal cord movement was limited at the injected side. At 3 months, movement of the bilateral vocal cords was normal, and the vocal cord process mucosa was smooth. Patients were followed up for more than a year, and only one patient had recurrence in 2 months after surgery. The cure rate was 96. 4%. CONCLUSION Combination of laryngeal microsurgery and type A botulinum toxin local injection can shorten the treatment course of vocal process granuloma.
Anesthesia, General ; Botulinum Toxins ; administration & dosage ; Granuloma ; drug therapy ; surgery ; Humans ; Injections ; Intubation, Intratracheal ; Laryngeal Mucosa ; Laryngeal Muscles ; Laryngeal Neoplasms ; drug therapy ; surgery ; Laryngoscopes ; Larynx ; Microsurgery ; Postoperative Period ; Recurrence ; Vocal Cords ; Wound Healing

Maximum a Posteriori (MAP) method has been widely applied to the ill-posed problem of image reconstruction. The choice of prior is the crucial point on MAP methods. However, the most conventional priors will lead to a blurring of the whole image or cause ladder-like artifacts. We therefore proposed a Tsallis entropy-based prior for positron emission tomography (PET) iterative reconstruction in MAP framework. The method uses a Tsallis entropy-based prior to eliminate the uncertainty between prior information and the estimated images. We tested this method in the phantom image, compared it with the traditional prior methods. the results showed that the proposed algorithm could suppress noise and obtain better reconstructed image quality.
Algorithms ; Artifacts ; Entropy ; Image Processing, Computer-Assisted ; methods ; Phantoms, Imaging ; Positron-Emission Tomography ; methods

OBJECTIVE: To investigate hypoxia tolerance of children with recurrent respiratory papillomatosis with the spread of trachea in surgical procedures without ventilation via endotracheal tube intermittently under general anesthesia. METHOD: Forty children with recurrent respiratory papillomatosis were enrolled in the observation. The duration of SpO2 from 100% to the points of 99%, 95%, 90%, 85%, the heart rate in each time point above. PaO2, PaCO2, pH valve when SpO2 was 85%, the duration of SpO2 back to 100% were recorded respectively. RESULT: Duration of SpO2 from 100% down to 99%, 95%, 90%, 85% was (168.4 +/- 58.3)s, (204.6 +/- 56.4)s, (224.8 +/- 58.9)s, (239.9 +/- 60.6)s, respectively. Heart rate was (121.6 +/- 14. 6)bpm, (123.3 +/- 15.1) bpm, (124.1 +/- 14.8)bpm, (125.0 +/- 15.1)bpm, respectively. When SpO2 was 85%, pH value was 7.22 +/- 0.05, PCO2 was (69 +/- 8.7)mmHg, PO2 was (52 +/- 7.9)mmHg. Duration of SpO2 up to 100% was (28.6 +/- 2.5)s; When SpO2 back to 100%, pH value was 7.40 +/- 0.02, PCO2 was (40.5 +/- 2.0)mmHg, PO2 was (358 +/- 104.3)mmHg. CONCLUSION Intermittent apnea during the surgical procedures in RRP children with distal spread of papillomas was safe, hypercapnia and hypoxia caused by the apnea can be corrected soon after the re-ventilation. Duration of apnea should be controlled within 3--5 minutes in each apnea-reoxygenation cycle.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Hypoxia ; physiopathology ; Male ; Monitoring, Intraoperative ; Papillomavirus Infections ; pathology ; surgery ; Respiratory Tract Infections ; pathology ; surgery ; Respiratory Tract Neoplasms ; pathology ; surgery