Background N-ethyl-N-nitrosourea (ENU)-induced mouse mutagenesis is a powerful approach for the study of gene function and the generation of human disease models.Objective This study was to create the corneal morphologic change and map the mutant gene of a kind of corneal opacity in ENU mutagenesis in mouse.Methods ENU was intraperitoneally injected in forty C57BL/ 6J (B6) male mice aged 8-10 weeks old.The male mice were mated with the same strain female mice.Their progenies were screened for visible eye mutation,and the mutant mice were mated with the same strain mice to confirm the heredity of mutation phenotypes.Hematoxylin & eosin staining was used to examine the histopathological change of cornea in one mouse with ENU-induced corneal opacity.To map the mutant gene,[(B6×D2)F1 ×B6] N2 mutant mice were bred,and the genome of the N2 mice was scanned by microsatellite markers distributed equally on the mouse chromosome.The microsatellite linked to the mutant gene was determined by the log odds score.This experimental procedure was approved by Ethic Committee about Experimental Animal Care and Use of Yangzhou University.Results The founder mouse,which was the progeny of an ENU-treated B6 male mouse and an untreated B6 female mouse,had a corneal opacity phenotype.After mating the mutant with B6 mice,19 of 59 descendants appeared corneal opacity phenotype.Thickening of corneal stroma,neoangiogenesis,infiltration of inflammatory cells and proliferation of fibroblasts were exhibited in cloudy cornea in ENU-induced mutated mice under the optical microscope.After linkage analysis between microsatellite markers and the mutant gene,the mutant gene was linked to D2Mi307,which was located at 63.42 cM.Three cases of 26 N2 mice underwent recombination with the LOD 3.79.The mutant gene associated with the cornea phenotype was located on chromosome 2.Conclusions This study map the mutant gene associated with the cornea phenotype on chromosome 2.The strain might be used as a mouse model for heritable human corneal opacity.

BACKGROUND: The effects of low-level laser biostimulation have been proved by various experimental studies and clinical application, which are manifested as tissue repairing, analgesia, antiinflammation, etc.OBJECTIVE: To observe the effects of He-Ne laser and low-level CO2 laser irradiation on fracture healing.DESIGN: A randomized controlled animal experiment. SETTING: Research Room of Laser Medicine, Chengde Medical College.MATERIALS: Forty-eight healthy male New Zealand rabbits, weighing 2.0-2.5 kg, were used.METHODS: The experiments were carried out in the Research Room of Laser Medicine, Chengde Medical College from 1998 to 2003. ①The 48 rabbits were induced into models of experimental fracture of the left radius, and then they were divided into three groups with 16 rabbits in each group: control group, He-Ne laser irradiation group and CO2 laser irradiation group. The fracture areas of the animals in the He-Ne and CO2 laser irradiation groups were respectively irradiated with 28 mW/cm2 He-Ne laser and 150 mW/cm2 CO2 laser for 10 minutes, once a day. ② The animals were killed on the 15th and 35th days after taking roentgenogram respectively. At 15 days, the collagen and calcium contents in callus were detected. At 35 days, the biomechanic anti-torsion strength was examined.MAIN OUTCOME MEASURES: ①Results of roentgenogram in each group; ② Collagen and calcium contents in callus; ③ Results of biomechanic anti-torsion strength.RESULTS: Totally 45 rabbits were involved in the analysis of results. ① On the 15th and day after fracture, the roentgenogram, collagen and calcium contents in callus in the He-Ne and CO2 laser irradiation groups were better than those in the control group, and the collagen and calcium contents in callus were higher in the CO2 laser irradiation group than in the He-Ne laser irradiation group [(341.9±30.1) vs (302.1±28.7) mg/g; (197.1±19.7)vs (156.5±17.6) mg/g, P＜ 0.05]. ②On the 35th days after fracture, the biomechanic anti-torsion functions in the He-Ne and CO2 laser irradiation groups were superior to that in the control group, and there were no significant difference between the two laser irradiation groups. ③ The results of roentgenogram in the He-Ne and CO2 laser irradiation groups were better than that in the control group both on the 15th and 35th days after fracture, and there was no significant difference between the two irradiation groups.CONCLUSION: Both CO2 laser and He-Ne laser irradiations can accelerate the process of fracture healing, and the curative effect of CO2 laser is better than that of He-Ne laser on the 15th day after fracture, but there is no significant difference clinically between the CO2 laser and the He-Ne laser irradiations on the 35th day after fracture.

Objective To investigate the safety and efficacy of a customized guide plate which is designed by digital medical technology and manufactured by 3D printing in clinical percutaneous needling at the hip joint.Methods Enrolled in this study were 9 patients with hip disorder who required hip joint needling during the period from April 2015 to August 2015.They were 7 males and 2 females,21 to 53 years of age (average,42 years).Laterality:5 left sides and 4 right sides.Conditions:4 cases of traumatic arthritis and 5 cases of osteoarthritis.All of them underwent lamellar CT scans whose data were imported into the software of Mimics 10.01 for 3D reconstruction.Then the software of 3-matic was used to design a customized guide plate on the reconstructed models.Finally,3D printing was used to manufacture the guide plate.The needle angle,direction and depth were adjusted in vitro and the operation was simulated before actual needling in clinic.After percutaneous hip joint needling was performed in the 9 patients separately,the accuracy of needling was checked using C-arm fluoroscopy.Results The percutaneous hip joint needling was performed successfully in the 9 patients in whom the customized guide plate was used.The results were consistent with what had been designed on the reconstructed models.The operation time in the 9 patients averaged 417.7 s (range,from 387 s to 590 s).Little intraoperative bleeding was observed at the site of needling,fluoroscopy was applied only once for check.No neurovascular injury or other operation-related complications occurred.Conclusion For patients who need repeated percutaneous needling at the hip joint,the customized guide plate designed by us can improve accuracy and ensure safety and efficacy of needling.

Objective To investigate the effects of Baicalin on intestinal mucosal injury in rats with partial common bile duct ligation (PCBDL).Methods Forty male Wistar rats were randomly divided into 4 groups equally:sham operation,PCBDL,PCBDL1 and PCBDL2.Rats in PCBDL,PCBDL1 and PCBDL2 groups were subjected to partial common bile duct ligation.Baicalin [80 mg/(kg · d)] was fed in PCBDL1 group (for 2 weeks) and PCBDL2 group (for 3 weeks),while for other groups,9 g/L saline in the same volume was fed.Ileum mucosa were prepared for microscopic examination.The intestinal mucosal injury in rats was observed and scored.The level of NF-κB mRNA expression by Fluorescent in Situ Hybridization,and the level of NF-κB protein were determined by immunohistochemistry.Results 1.Compared with PCBDL group (3.2 ± 0.5),the pathological severity scores of intestinal mucosa significantly declined (F =21.120,P ＜ 0.01) in PCBDL1 group (1.9 ± 0.2) and PCBDL2 group (1.5 ± 0.3).2.Compared with sham operation group(0.066 ± 0.006),PCBDL1 group (0.107 ± 0.011),and PCBDL2 group (0.098 ± 0.010),NF-κB expression in PCBDL group (0.155 ± 0.012) presented significantly up-regulation (F =76.8,P ＜ 0.01).3.Compared with sham operation,PCBDL1 group,and PCBDL2 group,the positive expression rates of NF-κB mRNA of intestinal mucosal epithelium in PCBDL group significantly increased.Conclusions It is suggested that Baicalin exert protective effects on the intestinal mucosal injury in rats with PCBDL,partially by inhibiting NF-κB mRNA,down-regulating NF-κB protein expression of intestinal mucosal epithelial cells.

Aim To investigate estradiol inhibition of neointimal proliferation after rat carotid artery balloon injury. Methods Eight to ten-week-old SD rats (male,n=21,female,n=21) were divided into intact control(n=7),gonadectomy control(n=7) and estradiol (n=7, gonadectomy)groups in each sex. Left carotid artery was not injured with 2.0 F PTCA balloon until estradiol was injected for three days. Rats were killed 2 wk after injury. Neointimal areas and media area, ratios of intimal areas/media areas were measured with computer. Results Male neointimal areas and ratios of intimal areas /media areas in estradiol group were less than those in intact control group significantly(all P0.05). Conclusions Estrdiol inhibits neointimal proliferation after the gonadectomy in rats undergoing carotid artery balloon injury.

Objective To study the capability of high field MRI in demonstrating the post-mortem fetal brains at different gestational age(GA).Methods One hundred and eight post-mortem fetal brains of 14-40 weeks GA were evaluated by 3.0 T MRI. Eleven brains of 14 to 27 weeks GA with good 3.0 T MRI images were chosen and scanned by 7.0 T MRI. The developing sulci, layered structures of fetal cerebral cortex and basal nuclei were evaluated on MRI of different Tesla(3.0 T and 7.0 T)and their results analyzed. Results On T_1 WI of 3.0 T MRI, the layered structures of fetal cerebral cortex were present at 14 weeks GA, the sulci were more accurately identified after 16 weeks GA. The basal nuclei were clearly distinguishable after 20 weeks GA. and these structures were better visualized as the GA increased. On T_2WI of 7.0 T MRI, the sulei, layered structures of fetal cerebral cortex and basal nuclei were shown more clearly at the same GA when compared to 3.0 T, especially the sulci at the early developmental stages. Conclusions T_1 WI of 3.0 T MRI could show the developing structures of post-mortem fetal brain well, but the T_2 WI of 7.0 T MRI were comparatively better.

BACKGROUND AND OBJECTIVEThe brain is one of the most common metastatic sites of breast cancer. Brain metastases develop in 10%-15% of patients with breast cancer and are associated with poor prognosis. The purpose of this retrospective study was to analyze the clinical characteristics and survival of patients with brain metastases due to breast cancer of different subtypes and to identify the prognostic factors that affect clinical outcome.

METHODSA total of 89 patients with breast cancer brain metastases diagnosed between October 1997 and July 2008 at Sun Yat-sen University Cancer Center were included in this study. Among the 89 patients, the number of luminal A, luminal B, human epidermal growth factor receptor 2 (HER-2), and triple-negative (TN) subtypes were 30, 20, 16, and 14, respectively; 9 patients had an unknown subtype. The clinical characteristics, pathologic features, and prognostic factors were analyzed both at the initial diagnosis and at the diagnosis of brain metastases. Endocrine therapy for patients with luminal subtypes was further studied.

RESULTSThe median age of patients was 46 years (range 28-74 years). The median survival time was 8.0 months (range, 0-80 months), the 1-year survival rate was 32% and the 5-year survival rate was 4%. The time to brain metastasis differed according to clinical stage at the initial diagnosis, and the time for patients with the luminal A subtype was the longest (P < 0.001). Multivariate analysis demonstrated that performance status score > 1, multiple brain metastases and without whole brain radiotherapy (WBRT) in combination with chemotherapy were associated with poor prognosis. Compared with the luminal A subtype, features of the HER-2 and TN subtypes included early metastases, rapid progression after first-line treatment (8.0 months vs. 11.0 months), and poor overall survival (25.0 months vs. 63.0 months). The luminal A subtype showed a tendency for good prognosis and slow growth. Tamoxifen could improve the survival of luminal A/B subtypes (median survival 24.0 months vs. 7.0 months, respectively, P = 0.002).

CONCLUSIONSThe prognosis of brain metastases from breast cancer was poor, especially in patients with HER-2 and TN subtypes. Generally, WBRT in combination with chemotherapy was the standard treatment modality. Patients with the luminal subtypes could benefit from tamoxifen.

Adult ; Aged ; Antineoplastic Agents, Hormonal ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Brain Neoplasms ; secondary ; therapy ; Breast Neoplasms ; classification ; pathology ; therapy ; Carcinoma, Ductal, Breast ; classification ; pathology ; secondary ; therapy ; Chemotherapy, Adjuvant ; Cranial Irradiation ; methods ; Female ; Follow-Up Studies ; Humans ; Mastectomy ; methods ; Middle Aged ; Neoplasm Staging ; Radiotherapy, Adjuvant ; Receptor, ErbB-2 ; blood ; Receptors, Estrogen ; blood ; Receptors, Progesterone ; blood ; Retrospective Studies ; Survival Rate ; Tamoxifen ; therapeutic use

OBJECTIVETo investigate the effects of recombinant BmKIM (poly-peptide derived from Asian Scorpion Buthus martensi Karsch) on the sodium current (I(Na)) of isolated ventricular myocytes, transmembrane action potential and aconitine induced arrhythmia in vivo in rabbits.

METHODSVentricular myocytes were enzymatically dissociated from adult rabbits. Whole-cell patch-clamp technique was used to record voltage-dependent I(Na). Standard transmembrane action potentials in rabbit hearts in vivo were recorded by using floating glass microelectrodes. Incidence of arrhythmias, the early after depolarization (EAD) and/or delay after depolarization (DAD) were measured in vivo in rabbits post aconitine (100 microg/kg, iv) in the absence or presence of BmKIM (50 microg/kg iv).

RESULTS(1) BmKIM significantly inhibited I(Na) in a voltage-dependent manner and significantly shifted the I-V curves of I(Na) upward. BmKIM left shifted the inactivation curve of I(Na) and voltages at 50% inactivation of I(Na) were changed from (-70.8 +/- 2.6) mV to (-84.8 +/- 3.5) mV (P < 0.05). BmKIM prolonged the recovery of inactivation of I(Na). In the presence of BmKIM, the time constants of recovery (both tau(f) and tau(s)) of I(Na) were significantly prolonged from (28.9 +/- 6.1) ms and (107 +/- 21.6) ms in control group to (54.2 +/- 7.9) ms (P < 0.05) and (211.1 +/- 34.6) ms (P < 0.01), respectively. (2) BmKIM significantly shortened 50% and 90% of action potential duration (APD(50) and APD(90)), and reduced action potential amplitude (APA), declined maximum up stroke velocity of action potential (V(max)) in vivo. The Q-T duration was shortened and heart rate significantly increased post BmKIM injection. (3) Incidence of aconitine induced ventricular arrhythmias (77.8%) was significantly reduced by BmKIM (22.2%, P < 0.01).

CONCLUSIONSBmKIM significantly blocked I(Na) through affecting the inactivated state of I(Na) in rabbit ventricular myocytes. BmKIM could attenuate the influx of I(Na), therefore shorten action potential duration and reduce action potential amplitude and reduce the incidence of aconitine induced arrhythmias.

Action Potentials ; Animals ; Anti-Arrhythmia Agents ; pharmacology ; Arrhythmias, Cardiac ; metabolism ; physiopathology ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Peptides ; pharmacology ; Rabbits ; Recombinant Proteins ; pharmacology ; Scorpion Venoms ; pharmacology ; Sodium Channels ; metabolism

Objective To study the developmental process of the region of basal nuclei of postmortem fetuses by 3.0 T and 7.0 T MRI.Methods One hundred and thirty-one postmortem fetuses of 14 to 40 weeks of gestational age(GA)were scanned by 3.0 T MR,of which 11 fetuses of 14-27 weeks of GA were chosen and scanned by 7.0 T MR. The time when the structures in the region of basal nuclei could be detected and the changes of MR signal intensity were analyzed for MRI of different Tesla.Results On 3.0 T MRI.the dorsal thalamus could be delineated as early as 14 weeks of GA. The germinal matrix, caudate nucleus,and putamen could be visualized as early as 15 weeks of GA. The globus pallidus could be described as early as 18 weeks of GA.and the internal capsule and external capsule could be shown as early as 20 weeks of GA. The signal of the caudate nucleus during 15-30 weeks of GA was relatively hypointense on T1WI and hyperintense on T2WI.but during 31-40 weeks of GA, it was relatively hyperintense on T1WI and hypointense on T2WI. The putamen had a relatively high signal intensity on T1WI and low signal intemity on T1WI during 15-17 weeks of GA, and it appeared patchy during 18-25 weeks of GA,then it had a relatively low signal intensity on T1WI and high signal intensity on T2WI during 26-30 weeks of GA, and during 31-40 weeks of GA,its signal intensity was relatively high on T1WI and low on T2WI.The globus pallidus had a relatively high signal intensity on T1WI and low signal intensity on T2WI during 20-40 weeks of GA Compared to the 3.0 T MRI,the T2 images of 7.0 T MRl were more clear,and most structures in the region of basal nuclei could be clearly displayed as early as 16 weeks of GA.such as the germinal matrix,caudate nucleus,dorsal thalamus,putamen,globus pallidus,internal capsule,and external capsule.The claustrum could be delineated as early as 18 weeks of GA on 7.0 T MRI. Conclusions 3.0 T MRI could show the developmental process of the region of basal nuclei well,but the T2 images of 7.0 TMRl were comparatively better.

OBJECTIVETo study the characteristics of thyroglossal tract cyst epidemiology and summarize how to improve the operation therapeutic effect.

METHODSThe clinic appearance, the pathological characteristics and the therapeutic effect of the 60 thyroglossal tract cyst cases were analyzed.

RESULTSThyroglossal tract cyst mostly occurred in children and teenagers. The incidence of female patients was more than that of male patients. Most of thyroglossal tract cyst located in the middle line of the neck and were explored tubular-objects and fibroropes. The histopathological feature was variform.

CONCLUSIONThe clinical appearance and pathology characteristic of thyroglossal tract cyst are variform. Thyroglossal tract cyst may be recurred easily if it is not operated well.

Adolescent ; Child ; Female ; Humans ; Male ; Thyroglossal Cyst