With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.
Glycoproteins ; chemistry ; Glycosylation ; Polysaccharides ; chemistry ; Ultrafiltration ; instrumentation

Objective To investigate the spherical diopter and astigmatism change of humans at sitting and supine position.De- sign Prospective case series.Participants 96 eyes of 52 patients (spherical diopter from-2.50 D to-10.00 D,astigmatism diopter from -0.75 D to-4.50 D) were selected.Methods The subjects were examined with NIKON portable retinomax at sitting and supine posi- tion,respectively.Main Outcome Measures The spherical diopter,cylinder diopter and axis change were analyzed statistically.Re- sults Spherical diopter at supine position (-5.31?3.43 D) was a little higher than that at sitting position (-5.27?3.24 D) statistically(P= 0.25),and cylinder diopter at sitting position (-2.27?1.24 D) and at supine position (-2.35?1.19 D) was no statistically difference (P= 0.20).The axis of astigmatism changed from-16?to +18?.Axis change was within 2?in 52.1% eyes,6?-10?in 5.2%,over 10?in 3.1%. The change of axis rotation tended to counter-clockwise in the right eye and clockwise in the left eye.Conclusions Eye rotation at sit- ting and supine position may cause the astigmatism axis change.It may be one of the main factors affecting the results of LASIK.

Objective To analyze the clinical manifestations and electroencephalogram (EEG)characteristics of agyria-pachygyria for its early diagnosis,treatment and prognosis judgment in clinical practice.Methods The clinical manifestations and EEG features of twenty-four patients with agyria-pachygyria who were diagnosed by CT or magnetic resonance imaging(MRI) at Pediatric Neurology of Children's Hospital of Chongqing Medical University from July 2004 to July 2013 were retrospectively analyzed.Results Of twenty-four patients,eighteen cases were diagnosed as diffuse agyria-pachygyria and six cases were diagnosed as partial agyria-pachygyria.The clinical features were mainly manifested as mental retardation (twenty-four patients),and motor retardation (twenty-four patients),and epilepsy (eighteen patients).All of the twenty-four patients had abnormal EEG pattern which were mainly three tapes.Type Ⅰ had diffused high amplitude alpha and beta activity in all cortical regions,frontal-central,or parietal-occipital region (fourteen patients).Type Ⅱ showed alternating high amplitude bursts with sharp and slow waves (seven patients).Type Ⅲ was characterized by high amplitude spike or sharp wave activity generalized or multifocal distribution and δ,θ wave mixing graphics (twelve patients).Nine of twenty-four patients showed two or three EEG characteristic patterns in an awake-asleep EEG recording.During the follow-up of 1-8 years old,twelve of the thirteen patients who were diagnosed as epilepsy in diffuse agyria-pachygyria had refractory epilepsy,mainly with infantile spasms or Lennox-Gastaut syndrome.One of the five patients who was diagnosed as epilepsy in focal agyria-pachygyria had refractory epilepsy,mainly for partial epilepsy secondary generalized seizures.There was a significant difference between them (P =0.008).Eighteen of twenty patients who had moderate-severe mental retardation or dyskinesia were diagnosed as diffuse a gyria-pachygyria,while two were focal agyria-pachygyria.Both of them had a significant difference (P =0.005).Conclusions Agyria-pachygyria is a brain malformation caused by neuronal migration abnormality.Diffuse agyria-pachygyria is presented with serious clinical manifestations and poor outcome while the clinical manifestation of focal agyria-pachygyria is relatively mild and epilepsy could be controlled by antiepileptic drugs or epilepsy surgery.These characteristics of EEG patterns along with clinical findings could provide important evidence for early diagnosis,timely treatment and prognosis judgment of agyria-pachygyria.

Objective To identify the relationship among serum 25-hydroxyvitamin D[25(OH)D] level, chronic inflammation state and malnutrition in maintenance hemodialysis (MHD) patients. Methods The serum 25 (OH)D level of 119 patients on MHD was detected. All the patients were divided into three groups according to the serum 25(OH)D level. Vitamin D deficiency: 25(OH)D ≤ 15 ng/mL, vitamin D insufficiency: 15 ng/mL <25(OH)D ≤ 30 ng/mL, vitamin D normal: 25(OH)D > 30 ng/mL. Inflammatory factor and nutritive index were compared among the three groups. Result The average of serum 25 (OH)D level was 3.4~45.3 ng/mL (22.5~14.8 ng/mL), and the prevalence of 25 (OH)D deficiency and insufficiency was 88.2%. Significant differences existed in age, inflammatory factor (IL-6, TNF-α, CRP) and nutritive index (Alb, PA, SGA) between 25(OH)D deficiency, insufficiency groups and normal group (P<0.05). Conclusion The serum 25(OH)D level is possibly related to the malnutrition in MHD patients. It may inhibit chronic inflammation state , in which it can promote state of nutrition of MHD patients.

Objective To investigate the clinical efficacy of mallet needle plus acupuncture in treating stiff neck. Method Eighty patients with stiff neck were randomly allocated to treatment and control groups, 40 cases each. The treatment group received mallet needle therapy plus acupuncture and the control group, acupuncture alone, for three consecutive days. In the two groups, the Pain Rating Index (PRI) score and the Visual Analogue Scale (VAS) score were recorded before and after treatment and the clinical therapeutic effects were evaluated after treatment. Result The PRI score and the VAS score decreased in both groups after treatment compared with before (P<0.05) and decreased more in the treatment group than in the control group at the end of treatment (P<0.05). The total efficacy rate was 95.0% in the treatment group and 87.5% in the control group; there was a statistically significant difference between the two groups (P<0.05). Conclusion Mallet needle therapy plus acupuncture is more effective than acupuncture alone in treating stiff neck.

Adolescent ; Adult ; Cross Infection ; microbiology ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Organophosphate Poisoning ; Pesticides ; poisoning ; Pneumonia ; microbiology ; Respiration, Artificial ; Respiratory Insufficiency ; chemically induced ; therapy

G mutation were intervened and avoided the occurrence of deafness,1 babies with 235delC homozygote was confirmed severe sensorineural hearing loss in the hearing screening.Conclusion Newborn gene screening make up the defects of missed diagnosis in simple hearing screening in finding the newborn babies with late-onset deafness or the high risk as well as the pathogenic carriers.So the hearing and gene screening were necessary in the current situation,and this screening strategy would be developed further in Henan province.

OBJECTIVETo study the influence of experimentally osteoporosis to osteogenic efficiency of bone matrix gelatin(BMG) implanted into the calvarial defects of rats.

METHODSSixty-eight female SD rats of 12 +/- 1 weeks were randomly divided into two groups with 34 rats in each group. The ovaries were excised in the ovariectomized group (VG). The control group underwent sham surgery. Ninety days after ovariectomy, 10 rats from each group were examined to ensure the formation of postmenopausal osteoporosis by measuring bone density of the femur with single photon absorptiometric measurements. A critical-sized (8 mm in diameter) calvarial defect was created on the rest of 48 rats. Bone matrix gelatin was implanted to the defect. The rats were scarified at the 21st and 56th day after surgery respectively. The new bone forming capability of BMG was evaluated with undecalcified histological observation, tetracycline fluorescence marker, quantitative bone histomorphometry, At 90th day after ovariectomy, bone density of scanning electron microscopy and X-ray spectrometry.

RESULTSOVG showed very significant difference compared with the control group (0.315 +/-.015) g/cm2 vs [(0.347 +/- 0.017) g/cm2, P < 0.01 ]. At the 21st day following the implantation operation, new bone formed within the bone defects in both groups. The amount of new bone in OVG was lower than the control group. The tetracycline-labeled region in the bone defect was sparser in the OVG. At the 56th day, the bone defects healed mostly in the control group but fibrous tissue filled parts of bone defect in the OVG. The distance between two fluorescent lines of incorporated tetracycline and the mean mineralization deposition were significantly lower in the OVG than the control at the 21st day and 56th day. Mineralization of callus in OVG was inferior. Significant difference was found between the OVG and the control group in the calcium to phosphate ratio of callus in bone defects at the two time-points.

CONCLUSIONSExperimentally induced osteoporosis depressed osteogenic efficiency of BMG, suggesting that estrogen could play an important role in bone remodeling with bone substitute participating.

Animals ; Bone Matrix ; chemistry ; Bone Regeneration ; Female ; Femur ; metabolism ; Gelatin ; therapeutic use ; Osteogenesis ; Osteoporosis ; surgery ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Skull Fractures ; surgery

ObjectiveTo study the intelligence quotient(IQ) profile of the children with idiopathic generalized tonic-clonic seizure (GTCS) and the factors influencing IQ of them.MethodsAll 28 children with GTCS were selected who were aged 9 ～ 16 years in 20 GTCS families in a mountain area of the south of Anhui Province,all available healthy siblings of the children included in the epilepsy group,did not have epilepsy,and were nearest in age to the children with GTCS ( control group 1 ),and thirty children aged 9 ～ 16 years old who had lived in the same village ( control group 2) entered into our study.The IQ was studied of the three groups of children and the factors influencing IQ of children with GTCS at the same time.The data were input SPSS12.0 and analyzed.ResultsThe IQ of children with GTCS(85.64 ±20.57)was lower than that control group 1( 103.39 ± 11.17)and the control group 2 ( 106.17 ± 11.67).The difference between children with GTCS and the two control groups were significant for almost all the subtest quotients except completion of drawing and picture arrangement.No significant differences were found between the control group 1 and the control group 2 on the IQ and the subtest quotients.IQ scores of children with GTCS showed significant linear correlation with father's education( r=0.453,P＜0.01 ),age at onset of epilepsy( r=0.506,P＜0.01 ),duration of seizure disorder( r=-0.533,P＜0.0l ),status epilepticus( r=-0.732,P＜0.01),total number of seizures( r=-0.761,P＜0.01) and seizure frequency ( r=-0.708,P ＜ 0.01 ).ConclusionThe IQ scores of the children with idiopathic GTCS are lower significantly than general children population.Epilepsy-related variables affecting IQ scores of the children with idiopathic GTCS are duration of seizure disorder,status epilepticus,age at onset of epilepsy,total number of seizures,seizure frequency.

Activation of acid-sensing ion channels (ASICs) plays an important role in neuroinflammation.Macrophage recruitment to the sites of inflammation is an essential step in host defense.ASIC 1 and ASIC3 have been reported to mediate the endocytosis and maturation of bone marrow derived macrophages.However,the expression and inflammation-related functions of ASICs in RAW 264.7 cells,another common macrophage,are still elusive.In the present study,we first demonstrated the presence of ASIC 1,ASIC2a and ASIC3 in RAW 264.7 macrophage cell line by using reverse transcriptase polymerase chain reaction (RT-PCR),Western blotting and immunofluorescence experiments.The non-specific ASICs inhibitor amiloride and specific homomeric ASIC 1 a blocker PcTx 1 reduced the production of iNOS and COX-2 by LPS-induced activating RAW 264.7 cells.Furthermore,not only amiloride but also PcTx 1 inhibited the migration and LPS-induced apoptosis of RAW 264.7 cells.Taken together,our findings suggest that ASICs promote the inflammatory response and apoptosis of RAW 264.7 cells,and ASICs may serve as a potential novel target for immunological disease therapy.