Neural cell adhesion molecule (NCAM) is a glycoprotein expressing on the surface of neurons, glial cells, bone cells and natural killer cells. NCAM plays an important role in the process of cell - cell adhesion and cell migration, and is also a model protein to study polysialic acid. In this paper, NCAM gene from mouse mammary gland cells (NMuMG) was cloned into eukaryotic expression vectors pcDNA3.1(+) and transfected into mutant Chinese hamster ovary cells ldlD-14. The stable transfection over-expressing NCAM was obtained through the G418 selection and confirmed by Western blotting. Due to unique characters of ldlD-14 cells, carbohydrate chain of NCAM molecule can be easily manipulated with or without adding galactose in the serum free medium, and this modification can provide the basis for further studies on the effect of glycosylation on NCAM molecular function.
Animals ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Female ; Galactose ; Glycosylation ; Mammary Glands, Animal ; cytology ; Mice ; Neural Cell Adhesion Molecules ; biosynthesis ; Polysaccharides ; chemistry ; Sialic Acids ; chemistry ; Transfection

OBJECTIVE:To study the ADR mechanism of conjunctival hyperemia in model rats with prostacyclin-induced high intraocular pressure. METHODS:50 rats were randomly divided into normal control group,model group,prostacyclin low-dose, medium-dose,high-dose groups(100,200,400 mg/kg),10 in each group. Except for normal control group,right eyes of rats in other groups were established high intraocular pressure model,dropping corresponding medicine once a day,for 1 week. After last administration,the right eyes cornel peripheral corneal endothelial cells of rats in each group were isolated in vitro and cultured. Vascular endothelial cell viability,cell apoptosis and proliferation-related factor(Ki-76),apoptosis-related factors(Bad,Bax),in-hibito of apoptosis-related factors (Bcl-2,Bcl-xl) protein expressions were detected. RESULTS:Compared with normal control group,vascular endothelial cell viability in model group were obviously decreased;apoptosis rate was obviously increased;Bad, Bax protein expressions were obviously enhanced;Bcl-2,Bcl-xl,Ki-76 protein expressions were obviously weakened,with statisti-cal significances (P<0.05 or P<0.01). Compared with model group,vascular endothelial cell viability in prostacyclin low-dose,medium-dose,high-dose groups were obviously decreased;apoptosis rate was obviously increased;Bcl-2,Bcl-xl protein expres-sions were obviously weakened,with statistical significances(P<0.05 or P<0.01);the Bad,Bax protein expressions in prostacy-clin medium-dose,high-dose groups were obviously enhanced(P<0.05 or P<0.01),the other indexes had no statistical differenc-es(P>0.05). CONCLUSIONS:Prostacyclin may cause conjunctival hyperemia through promoting the apoptosis of cornel peripher-al corneal endothelial cells of model rats with high intraocular pressure and decreasing the cell viability.

Objective To study the relationship of microinflammation,nutrition and common carotid artery intima-media thickness(CCA-IMT) for the early prevention and interference of ischemic stroke. Methods(CCA-IMT was) measured by carotid arterial ultrasound in 250 elderly subjects.The levels of serum high-sensitivity C-(reactive) protein(hsCRP),ferritin,albumin,pre-albumin and transferrin were assayed at the same time.According to the results of CCA-IMT,all the subjects were divided into five groups:

Background The dysfunction of the blink reflex the eyelid-closure ability appears in the patients with facial paralysis,and its management is the implantation of mechanical-assisted eye-closure device in the upper eyelid.A novel device is palpebral spring implant.However,there is no similar study in China.Objective This study was to evaluate the clinical efficacy of palpebral spring placement for lagophthalmos caused by facial nerve palsy.Methods This clinical research complied with Helsinki declaration and the protocol was approved by Ethic Committee of Henan Eye Institute & Henan Eye Hospital.Written informed consent was obtained from each patient prior to the surgery.A retrospective serial case-observational study was performed.The medical records of 11 patients who underwent palpebral spring placement for hypophasis due to facial nerve palsy were reviewed at Henan Eye Hospital from August 2010 to November 2012.Palpebral spring placement was performed by the same surgeon to ensure a more even outcomes.Palpebral spring was made by nickel wire,with the diameter of 0.3 mm and implanted on tarsal plate in 11 eyes of 11 patients with symptomatic facial nerve palsy.The lower tip of Levine spring was encased into a small terylene bag and sutured to the anterior tarsal surface during the surgery.Preoperative and postoperative symptoms,upper eyelid margin to mid pupil distance (ULMD),degree of lagophthalmos and eyelid moving scope were examined and compared between before and after operation.The operating complication was followed-up for 8-38 months.Results The discomforted symptoms disappeared in all the operated eyes.The ULMD was (3.51±0.73) mm in preoperation and (3.20±0.86) mm in posteration,without significant difference between them (t=1.36,P=0.10).The degree of lagophthalmos was (5.94±1.57) mm and (1.06±0.98) mm in preoperation and postoperation respectively,showing a significant difference between them (t =9.42,P =0.00).The eyelid moving scope was (5.89±0.70) mm in postoperation,which was significantly increased in comparison with (0.11 ±0.33) mm of preoperation (t =22.97,P =0.00).The palpebral spring implant was regulated in 1 patient during the follow-up duration due to the trauma.No complication in other 10 patients appeared during the follow-up duration,such as implant exposure,metal fatigue or infection.Conclusions Palpebral spring placement is safe and effective for lagophthalmos in patients with facial nerve palsy.

Objective To observe the expression of synaptotagmin I(syt I)protein in the prefrontal cortex of adult-onset hypothyroidism rats and the effects of replicated therapy in different doses of thyroid hormone on the syt I protein.Methods All 44 aduh male Sprague-Dawley rats were divided into 4 groups randomly according to their body mass:hypothyroidism group,routine dosage thyroxine treatment group,high dosage thyroxine treatment group and control group.The adult male Sprague-Dawley rats were replicated to the adult-onset hypothyroidism and treatment models with propyhhiouracil(PTU).The levels of serum T3,T4 were assayed by the radioimmunoassay method and the level of the syt I protein in the molecular layer,external granular layer,external pyramidal layer,internal granular layer and internal pyramidal layer in prefrontal cortex was analyzed by immunohistochemistry.Results In the hypothyroidism group,the levels of serum T3 and T4[(0.34±0.04),(43.01±2.95)nmol/L]were significantly lower than those in the control group[(0.65±0.15), (55.20±3.56)nmol/L, F value: 6.026,5.940,4.503,P<0.05 or <0.01 ], the levels of the syt I protein in the molecular layer(0.018±0.010), external granular layer (0.020±0.007), external pyramidal layer(0.013±0.008), internal granular layer(0.011±0.005), internal pyramidal layer(0.024±0.013) of prefrontal lobe were significantly lower compared to the control group[(0.028±0.010,0.031 ± 0.010,0.028 ± 0.010,0.022 ± 0.008,0.038 ± 0.013), F value: 5.697,8.965,14.668,13.597,6.807,P<0.05 or <0.01 ]. In the routine dosage of the thyroxine treatment group, the levels of serum T3,T4 [(0.63 ±0.05), (55.04 ± 3.77)nmol/L] were not significantly different compared to the control group(F value: 3.162,0.367,all P>0.05), and the level of the syt I protein in the molecular layer, external granular layer, external pyramidal layer, internal granular layer and internal pyramidal layer in prefrontal cortex showed a significant improvement of the syt I protein(0.027 ± 0.013,0.025 ± 0.009,0.022 ± 0.008,0.020 ± 0.010,0.033 ± 0.010), which were similar to that of the control group(F value: 0.094,2.208,2.467,0.350,0.693, all P>0.05). In the high dosage thyroxine thyroid hormone treatment group, the levels of serum T3 and T4[ (1.11 ± 0.10), (96.68 ± 6.42)nmoL/L] were higher than the control group(F value: 6.291,12.031, all P<0.01), the expression of the syt I protein(0.028 ± 0.008,0.031 ±0.011,0.026 ± 0.012,0.023 ± 0.011,0.038 ± 0.010) were not significantly different compare to the control group (F value: 0.001,0.019,0.111,0.061,0.001, all P>0.05). Conclusions The expression of the syt I protein in the prefrontal cortex of adult-onset hypothyroidism can be decreased, which can be reversed by routine dosage of thyroxine treatment.

Objective To explore the technology and curative effect of thoracic endovascular aortic repair (TEVAR) for Stanford type B aortic dissection.Methods The clinical data of 85 patients with Stanford type B aortic dissection,who were admitted to authors' hospital during the period from January 2010 to April 2016 to receive TEVAR,were retrospectively analyzed.Conventional left brachial artery puncture and straight incision of right femoral artery were employed in all 85 patients,and DSA of ascending aorta was performed to find out the position of rupture,the position of the true and false lumens,and their relationship with the vascular openings of important organs.Endovascular covered stent was implanted to seal off the primary rupture;reexamination of ascending aorta angiography was adopted to check the sealing-off condition of the proximal rupture and the changes of blood flow in the aortic branches as well as in the true and false lumens.Results Successful TEVAR was accomplished in 84 patients.One patient died of sudden rupture of aortic dissection during preoperative anaesthesia.The technical success rate was 100％.In 9 patients the covered stent partially overlapped the left subclavian artery,in one patient the left subclavian artery “chimney” stem completely obstructed both the left common carotid artery and the left subclavian artery,and bypass surgery between left common carotid artery and left subclavian artery was carried out in 2 patients.After the treatment,internal leakage of type Ⅰ was detected in 2 patients.No death occurred during hospitalization period.After the surgery the patients were followed up for 3 months to 3 years,and all patients survived.New rupture at the distal site occurred in 2 patients.Conclusion For the treatment of Stanford type B aortic dissection,TEVAR is safe and effective.Strict observance of surgical indications,careful operative manipulation,and strengthening postoperative management after discharge from hospital are the key points to ensure a successful surgery as well as to improve the long-term survival rate.

OBJECTIVE:To establish a method of microbial limit test for liquid preparation of Iron sucrose injection before filtration and sterilization.METHODS:According to the microbial limit test in the 1005 and general rules 1006 of 2015 edition of Chinese Pharmacopoeia (vol.Ⅴ),plate method and membrane filtration method were used to measure total number of aerobic bacteria (Staphylococcus aureus,Pseudomonas aeruginosa,Bacillus subtilis,Candida albicans,Aspergillus niger) and total number of molds and yeasts (C.albicans,A.niger).The optimal test method was obtained by comparing the bacterial recoveries.RESULTS:By plate method,the recoveries of P aeruginosa and B.subtilis were 2％ and 5％.The test sample was diluted 10 times with pH 7.0 sodium chloride-peptone buffer solution,and the bacterial recoveries were in the range of 88％ to 96％;but he medium was dark in color.By membrane filtration method,without rinse solution,the bacterial recoveries in the range of 88％ to 95％.Add rinse solution,the bacterial recoveries were in the range of 91％ to 103％.After validated,the recoveries of menbrane filtration method with tlushing fluid ranged 50％-200 ％,which was in line with the requnements.CONCLUSIONS:The membrane filtration method established in this experiment has higher bacterial recovery rate than the plate method.The bacterial recoveries rate were higher after adding rinse solution,and no dark substance in the surface of filter membrane affect the accotmt.It can be used as the microbial limit test method for preparation liquid of Iron sucrose injection before filtration and sterilization.

Animals ; Depression ; physiopathology ; therapy ; Electric Stimulation Therapy ; methods ; Frontal Lobe ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism ; Transcranial Magnetic Stimulation ; methods

Objective To summarize clinical outcomes of interventional therapy on children with common congenital heart diseases(CHD).Methods A retrospective study was conducted.One hundred and fourteen patients with CHD were selected as our subjects,who underwent catheter interventional therapy in the Second Affiliated Hospital of Zhengzhou University from Jan.2004 and Dec.2012.The size of occluder was chose according to intraoperative echocardiography or cardiac imaging measurements,and occluder was released under assisted monitoring by subtraction angiography or cardiac ultrasound.Results There are all together 112 patients got the therapy successfully,2 cases failed(occluder detachment),and the success rate of operation was 98.2％.After the success of interventional,echocardiography examination showed that 11 cases were with star point across shunt,but the function of the around valve was not affected.One months after operation,echocardiography examination showed star point across shunt of 11 cases were disappeared,and no occluder was shifted as well as no thrombosis formed.Three months after operation,chest radiograph showed pulmonary congestion decreases and heart shadow was shrink.Thirty-eight cases were with three tricuspid regurgitation before operation and 32 cases were without reflux at 3 months after operation,and 6 cases relieved significantly.The patients were followed up for 6 months or 3 years,activity endurance was significantly improved than that before operation.No occluder was shiftand hemolysis and arrhythmia occurred.Meanwhile,No thrombosis or embolism occurred.Conclusion Interventional treatment for children with congenital heart disease is proved as a safe,effective methods and it have broad prospects in clinical application.