Objective To construct the eukaryotic expression vector of Notch1intracellular domain,p3XFLAG?CMV7?NICD1,so as to prepare for the further research and exploration of effect of Notch1 on promoting epithelial?mesenchymal transition of human lens epithelial cells. Methods The cDNA fragment was reversely transcribed by RT?PCR from total RNA extracted from the SRA01/04 cells and was encoded with the specific am?plification?targeted NICD1was obtained from the SRA01/04 cells,then the cDNA fragment was inserted into p3XFLAG?CMV7 to transcribe Esche?richia coli DH5α. And the recombinant plasmid was extracted after bacterial screening by LB plating medium and confirmed by the restriction endo?nuclease digestion and DNA sequencing. Results The target gene obtained had the same molecular size as predicted. It was indicated that recom?bined p3XFLAG?CMV7 plasmid contained correct recombinant human Notch1 sequences and p3XFLAG?CMV7?NICD1 was constructed success?fully. The western blotting showed protein NICD1 expressed in SRA01/04 cells transfected with p3XFLAG?CMV7?NICD1. Conclusion The suc?cessful construction of p3XFLAG?CMV7?NICD1 will provide a foundation for a further study studies in on the effect relationship of Notch signaling pathway and in posterior capsular opacification(PCO)after cataract extraction.

The methods of diagnostic imaging for pulmonary embolism is diversified. However, the different characteristic features of diagnostic imaging and diagnostic accuracy impressive influenced the choice of selection by clinicians. Furthermore, the principle and indication of interventional therapy would have great impending force on the outcomings. This article presents a comprehension of diagnostic imagings and interventional therapy for pulmonary embolization.

Objective:To investigate the common reasons for the failure of radical mastoidectomy in order to improve the result of treatment and obtain a dry ear.Method:Twenty-eight cases,who achieved no dry ear after radical mastoidectomy,underwent secondary surgery.Result:All cases obtained dry ear without vertigo or facial paralysis after operation and postoperative dressing.Conclusion:The reasons for the failure of radical mastoidectomy result from the incomplete clearance of lesions, the insufficient ventilation of mastoid cavities, the inappropriate postoperative dressings or the residual foreign bodies in surgical cavity.It is the key points to achieve skeletonization adequately, to eliminate the pathological tissues thoroughly under microscope, and to ensure unobstructed drainage of surgical cavities for preventing secondary surgery.

Objetive To study the medical value of the combined treatment of laparoscopy and duodenoscopy treatment for calculus of extrahepatic bile duct.Methods Retrospectivly analyzed the clinical data of 72 cases with combination of laparoscopy and duodenoscopy treatment for calculus of extrahepatic bile duct between March 2006 and October 2011.There were 11 cases of choledocholithiasis and 61 cases of choledocholithiasis and cholecystolithiasis.Results Three cases of severe abdominal adhesion converted to laparotomy and other 69 cases were cured.The cure rate was 95.83％.There were no complications such as infection,bile duct injury,bile leakage,intraabdominal hemorrhage,bile reflux cholangitis,pancreatitis and cicatrical stenosis of papilary.The examination after operation showed that there weren't any residual bile duct stones.Conclusions The combination of laparoscopy and duodenoscopy treatment for calculus of extrabepatic bile duct has less traumatic,few intercurrent disease,and was safe and reliable with well postoperative recovery.It can be regarded as the preference treatment for calculus of extrahepatic bile duct.

Hepatocyte transplantation is effective in the treatment of liver failure caused by a variety of factors.In pace with the progress of the study of induced pluripotent stem cells and its differentiation technology,a new method has arisen to obtain a great number of safe hepatocytes with biological function,which are suitable for seed cells of hepatocyte transplantation.In this article,we review the latest research progress about induced pluripotent stem cell-derived hepatocytes being transplanted to treat liver failure.

Objective To investigate the influence of Liuwei Dihuang pills on proliferation and differentiation of rat preadipocytes. Methods Rat preadipocytes were cultured, the influence of Liuwei Dihuang drug-containing serum on proliferation of rat preadipocytes was detected by MTT method, the influence of Liuwei Dihuang drug-containing serum on expression of GPDH was determined by enzyme tissue chemical method, the effect on lipid accumulation was determined by Oil Red O staining. Results Liuwei Dihuang drug-containing serum stimulated rat preadipocyte proliferation, inhibited GPDH up-regulation during differentiation, inhibited lipid accumulation in cell differentiation with the concentration below 10%. Conclusion Liuwei Dihuang pills stimulated rat preadipocyte proliferation, inhibited rat preadipocyte differentiation.

Asteroid Hyalosis (AH) is a common clinical disease,which has been considered a benign disorder as it rarely impairs visual acuity.It was often discovered when the patient was treated for other eye diseases.The mechanism was unclear.Its characteristic B-ultrasound property makes the B-ultrasound a very helpful diagnostic technique.In the case of the patients with other fundus diseases associated with AH,optical coherence tomography (OCT) and fluorescein angiography (FA) may be used to reduce the interference from asteroid bodies,therefore improve the fundus visibility.Recent studies have shown that AH can incorporate with many other eye diseases.For example,in patients with cataracts,asteroid hyalosis can cause surface calcification of silicone plate intraocular lenses,which in most cases may lead to the need for explantation of the calcified intraocular lenses.The efficacy of pars plana vitrectomy (PPV),the removal of some,or all,of the eye`s vitreous humor for AH remains controversial.In this paper,we provide a review of the recent literature on AH disease: the etiology,diagnosis and treatment.We hope to thus improve the awareness and outcomes of AH disease.

BACKGROUND:Ginsenoside can promote wisdom,prevent aging,protect cortical motor neurons,resist cell apoptosis,but the mechanisms are unclear.OBJECTIVE:To observe the effect of Ginsenoside Rb1 and Rg1 on nerve growth factor expression in Schwann cells.DESIGN,TIME AND SETTING:The in vitro cytological study was performed at the Second People's Hospital of Shenzhen City from March to June 2004.MATERIALS:Fresh adult ex vivo nerve was obtained from limbs that were dissociated by trauma and could not be reimplanted at the Second People's Hospital of Shenzhen City.Ginsenoside Rb1 and Rg1 was supplied by the Norman Bethune University of Medical Sciences.METHODS:Epineurium was removed and cut into 1.0-2.0 mm blocks.Schwann cells were isolated by enzyme digestion.Following removing fibroblasts by double 30-minute differential attachment,Schwann cells with above 95% purity rate were harvested,and then incubated on a 96-well culture plate coated with polylysine (105 cells/well).Schwann cells in the Ginsenoside Rb1 group were subjected to 20 uL of Ginsenoside Rb1 at 10,20,40,60,80 ug.Schwann cells in the Ginsenoside Rg1 group underwent 20 uL of Ginsenoside Rg1 at 10,20,40,60,80 ug.Schwann cells in the control group were treated with 20 uL of phosphate buffered saline.MAIN OUTCOME MEASURES:Nerve growth factor expression rate was determined in Schwann cells by using flow cytometry.RESULTS:Nerve growth factor expression rate in Schwann cells was significantly increased in the Ginsenoside Rb1 and Ginsenoside Rg1 groups compared with the control group at 48 hours following incubation (P ＜ 0.05),in a dose-dependent fashion.Nerve growth factor expression rate peaked when Ginsenoside Rb1 and Ginsenoside Rg1 were 60 mg/L.No significant difference in nerve growth factor expression rate was detected between the Ginsenoside Rb1 and Ginsenoside Rg1 groups (P ＞0.05).CONCLUSION:Ginsenoside Rb1 and Rg1 has potential of promoting the recovery of damaged peripheral nerve by increasing Schwann cell producing and secreting nerve growth factor.

BACKGROUND: Schwann cell derived neurotrophic factor, which is isolated and purified from the kytoplasm of Schwann cell with the relative molecular mass of 58000, is a kind of neurotrophic substance possessing obvious neurotrophic activity. It can be against neurovirulent substance of nitrogen monoxidum.OBJECTIVE:To create root avulsion animal models and observe the protective effects of Schwann cell derived neurotrophic factor (SDNF) on motoneurons of spinal anterior horn from spinal root avulsion induced cell death.DESIGN: Repeated observation and measure.SETTING: Third Department of Orthopaedics, Second People's Hospital of Shenzhen; Department of Micro-surgery , First Hospital Affiliated to Sun Yat-sen University.MATERIALS: This experiment was conducted at the Experimental Animal Center of Medical College of Sun Yat-sen University from March to May 2003. Twenty Sprague-Dawley rats with the age of 3-4 months, of clean degree, were selected and divided randomly into experimental group of Schwann cell derived neurotrophic factor and control group of normal saline with 10 rats in each group. The right side was injured, and the left side was intact served as normal control side.METHODS : ①A rat model of C6,7 spinal root avulsion induced motoneuron degeneration was established. ② A small piece of gelfoam presoaked in 40 μL SDNF solutions (1 g/L) was placed in contact with the injured spinal cord in the animals of the experimental group. Normal saline was added as the same way as above in the animals of the control group. ③ A silica pipe was put on the surface of gleform, one end of the silica was sutured to the glefoam , and the other end wasfixed subcutaneously with vaselinum. Local intramuscular injection of penicillinum was performed on the wound following closing the incision. All rats received an injection (20 μL) of either SDNF or normal saline solution at the lesion site through the silica pipe sutured to the glefoam once a week after the surgery. All the animals were killed by the end of the third weeks. ④The spinal region of C6,7 level was dissected out for observing survival rate and morphological change of motoneurons of spinal anterior horn as well as the expression of nitricoxide synthase(NOS).MAIN OUTCOME MEASURES: ① Survival and morphological change of spinal motor neurons. ②Change of nitricoxide synthase expression of spinal motor neurons.RESULTS: Totally 20 rats were enrolled in the experiment, and all of them entered the stage of result analysis. ① Survival and morphological changeof spinal motor neurons: 68.6% motoneurons of injured side of the control group died at 3 weeks after surgery. The survival rate was 31.4%,which was significantly lower than that of the intact side (P ＜ 0.01), and the survived neurons was shrinked significantly; the death rate of spinal motor neurons of injured side of experimental group was decreased by 35%as compared with control group (P＞ 0.05). The survival rate was 66.4%,and the survived neuron body was increased, similar to the intact side (P ＞ 0.05). ② Change of nitricoxide synthase expression of spinal motor neurons: In normal spinal cord, NOS positive neurons were shown in dorsal horn, surrounding the central canal and in the intermediolateral column.NOS was not seen in the anterior horn motonurons. At the end of the third week after C6,7 spinal root avulsion, increased NOS expression was not found at the injured side in the Schwann cell derived neurotrophic factor group and the intact side in the control side, while the significantly increased NOS expression of spinal motoneurons was found at the injured side of the control group.CONCLUSION: Degeneration of spinal motoneuron and increased expression of NOS can be induced by spinal root avulsion. SDNF has a significant effect in protecting spinal motoneurons from spinal root avulsion induced cell death and inhibiting the expression of NOS. These results suggest that the effects .of SDNF on motoneuron survival may be achieved by modifying the expression of certain cellular molecule such as NOS.

BACKGROUND: Schwann cell-derived neurotrophic factor is a bioactive protein isolated and purified from the kytoplasm of Schwann cell. It can obviously maintain the survival of spinal cord anterior horn motor neuron and promote the regeneration of peripheral nerve.OBJECTIVE: To observe the protective effect of Schwann cell-derived neurotrophic factor on the high injury of peripheral nerve-induced apoptosis of sensory neurons in spinal dorsal root ganglia.DESIGN: Randomized and controlled animal experiment.SETTING: Shenzhen Second People's Hospital.MATERIALS: Totally 30 3-week-old SD infant rats, of clean grade and either gender, were used in this experiment. They were randomly divided into neurotrophic factor group and control group with 15 rats in each one.Left sides of the animals in both two groups were set as normal sides and right sides as injured sides.METHODS: This experiment was carried out at the Experimental Animal Center, Medical College of Sun Yat-sen University from May 2003 to July 2003. ① L4.5 nerve root high-mutilation animal models were developed on the rats in two groups. Proximal nerve stump was connected with silicone tube. According to grouping, 60 mg/L Schwann cell-derived neurotrophic factors and 20 μL normal saline were injected into the silicone tubes respectively. Two ends of silicone tube were enveloped with vaseline.② Sample collecting was conducted at postoperative 4 weeks, survival rate and morphological change of sensory neurons in dorsal root ganglia of injured nerve was observed.MAIN OUTCOME MEASURES: ① Gross observation of sciatic nerve regeneration at injured side of the rats in two groups ② Survival of sensory neurons in dorsal root ganglia ③ Morphological change of sensory neurons in dorsal root ganglia.RESULTS: All the 30 rats entered the stage of result analysis. ① Gross observation of sciatic nerve regeneration: In the neurotrophic factor group,nerve new born axon grew along silicone tube, with 1cm in length; there were few and thin newborn axons in control group with 0.8 cm in length.② Survival of neuron in dorsal root ganglia of the rats in two groups: There was little fibrous tissue proliferation in the dorsal root ganglion in neurotrophic factor group. The loss of neurons was not obvious and the survival rate was 91.8%. Obvious fibrous tissue proliferation appeared in the dorsal root ganglia in control group, and a great many neurons were lost with the survival rate of 58.6%. Survival rate of neurons was 33.2% higher in neurotrophic factor group than in control group (P ＜ 0.01 ). ③ Morphological change of neurons in dorsal root ganglia: The diameter and area of neurons in dorsal root ganglia were significantly lower in control group than in neu rotrophic factor group and normal side [(21.8±1.4) μm,(373.1±50.9) μm2 vs (24.8±1.1) μm, (482.8±42.2) μm2 and (24.5±1.3) μm, (471.5±51.4) μm2,P ＜ 0.01], while there were no significant difference in diameter and area of neurons between neurotrophic factor group and normal side(P ＞ 0.05).CONCLUSION: Schwann cell-derived neurotrophic factors have obvious neurotrophic bioactivity for sensory neurons in the injured dorsal root ganglia.