Most of the proteomics analysis methods based on tandem mass spectrometry rely on the matching scoring of actual spectrum and theoretical spectrum, the interference of a large number of co-eluting peptides could cause error in the identification and quantification of peptides and proteins. Peptide retention time prediction, as a auxiliary and verification index of the peptide, can transition the chromatographic behavior into stable independent time attributes, and improve the accuracy of the peptide identification. Prediction of peptide chromatographic retention in complex systems is also of great significance for optimizing proteomics determination conditions and improving the detection rate and repeatability of mass spectrometry data in data-independent acquisition. This review focused on the chromatographic retention prediction method of unmodified peptides and modified peptides, summarizes the content, characteristics and limitations of four types of peptide retention time prediction methods based on standardized indexes, peptide molecular models, amino acid residue parameters, and machine learning, as well as their applications in proteomics, with a prospect of their future.

Objective To investigate the location of receptor interacting protein 3( RIP3) in Necroptosis and its function in this signal passage, and explore the relationship between receptor interacting protein 1 ( RIP1 ) and RIP3 in nuclear translocation. Methods Primary cerebrocortical neurons were cultured for 12 days,then pre-treated with zVAD-fmk(20μ,mol/L) for half an hour to block apoptosis. ①Extracting nuclear and cytoplasmic protein after neurons were exposed to TNF for different time ,then protein levels of RIP3 were analyzed by western blot and immunofluorescence for qualitative observation;②In the following research,the neurons were treated with Nec-1 and shRlPl ,then the protein level of RIP1 and RIP3 with western blot were analyzed, cell viability were determined by measuring LDH levels. Results ①In signaling pathways of necroptosis, the protein level of RIP3 in cytoplasmic decreased gradually with prolonged TNF exposure, to the corresponding it rolled up in nucleus and a-chieved the peak in 12 hours of TNF treatment ( Cytoplasmic 0. 45 ± 0. 03 ,0. 41 ± 0. 02,0. 73 ± 0. 03 ,0. 90 ± 0.01,1.15 ±0.04,1.30 ±0.02,0.99 ±0.03,0.63 ±0. 03;Nucleus 0. 07 ±0.02,0. 26 ±0.02,0. 57 ±0. 02,0. 68 ± 0.02,0. 80 ± 0.01,0.92 ± 0.02,1.28 ± 0.03,0. 87 ± 0.02) (P < 0.01). ②Blocking the relationship between RIP1 and RIP3 with necrostatin-1 and shRIPl , nuclear translocation of RIP3 decreased and caused a great increase in cell viability( 1.00 ±0.05,0.39 ±0.03,0.50 ±0. 03) (P<0. 01). Conclusion RIP3 mainly locates in cy-tolymph of normal cells,it translocates into nucelus as necroptosis takes place. RIP1 function with RIP3 in nuclear translocation. Block nuclear translocation of RIP3 is a potential way to protect cells.

Objective To explore the significance of NK cell activity,interleukin-2 receptors (sCD25) and glycosylated ferritin in the early diagnostic of acquired hemophagocytic lymphohistiocytosis (HLH).Methods 57 patients suspected of HLH from June 2005 to May 2008 and 25 healthy subjects were enrolled in the study.The patients suspected of HLH were divided into three groups i.e.(1) a group with diagnosis confirmed at first visit;(2) a group with diagnosis confirmed at subsequent visit and (3) a group with diagnosis unconfirmed according to HLH-2004 diagnostic criteria.Healthy subjects were enrolled as control.NK cell activity was determined with a released LDH assay.The percentage of glycosylated ferritin was determined with phytohemagglutinin adsorption assay,sCD25 was examined with ELISA double antibody sandwich assay.We compared the coincidence of each diagnostic index before and after diagnosis.Results The median percentage of NK cell activity was significantly lower in the first group ( 18.3±5.6) % and the second ( 16.7±6.7)% than that in the third group (33.4±6.8)% or in the controls (36.6±5.0)%.The median percentage of glycosylated ferritin was also significantly lower in the first group ( 15,4 ± 2.0)% and the second group (16.9 ± 3.4)% than that in the third group (40.4 ± 3.0)% or in the controls (45.2±2.2)%.Meanwhile,the median level of sCD25 was significantly higher in the first group (12 916±4328) ng/L and the second group (12 117 ± 5465) ng/L than that in the third group (4728±1482) ng/L or in the controls (3841 ± 993) ng/L.Furthermore,NK cell activity,sCD25 and glycosylated ferritin were abnormal in all the patients in the early stage of HLH.Conclusion NK cell activity,sCD25 and glycosylated ferritin may be helpful markers for the early diagnosis of HLH.

Objective To investigate the malnutrition status among the elderly in age care institutions in Shanghai suburb and analyze its potential influential factors.Methods The MiniNutritional Assessment (MNA) was adopted to evaluate the nutritional status of the 190 elderly people in age care institutions.The dietary supply by the institution canteen and the quantity of residual food left by the malnourished elderly people were weighted.Results In the age care institutions,the malnutrition rate reached 23.7％,47.9％ of the elderly people were at the risk of malnutrition,and only 28.4％ of the elderly people were well nourished.Logistic regression analysis showed that the major influential factors for malnutrition in the elderly people were food intake ability,mobile capability,chewing and swallowing ability,ageing and mental Illness.The malnourished elderly people had the most residual meat and vegetables,and insufficiency of nutrient intake was the main cause for the malnutrition in the elderly people.Conclusions The elderly people in age care institutions in Shanghai suburb have the higher risk of malnutrition.The malnutrition occurs under influence of many factors,of which some are unavoidable,however,some factors like dietary factors can be changed to improve the nutritional status of the elderly people in age care institutions.

Objective Studies show that the ERCC1 gene may be involved in secondary cisplatin resistance.This article aims to investigate the effects of shRNA targeting silencing excision repair cross-complementation group 1 (ERCC1-shRNA) on the proliferation and apoptosis of lung cancer A549/DDP cells treated with different concentrations of cisplatin.Methods Lung cancer A549/DDP cells were divided into a negative control, a blank control, an ERCC1-shRNA1, and an ERCC1-shRNA2 group.Human interfering RNA (RNAi) targeting the human ERCC1 gene was constructed and transfected into the A549/DDP cells using Lipofectamine 2000.The mRNA and protein expressions of ERCC1 in the A549/DDP cells were detected by real-time PCR and Western blot respectively, the proliferation-inhibition rate was assessed by MTT, and their cell cycle and apoptosis were determined by flow cytometry.Results ERCC1-shRNA was successfully constructed and transfected into the A549/DDP cells.Both the mRNA and protein expressions of ERCC1 were significantly lower in the ERCC1-shRNA1 (0.20±0.04 and 0.24±0.10) and ERCC1-shRNA2 (0.47±0.28 and 0.37±0.11) than in the negative control (0.96±0.12 and 1.32±0.13) and blank control groups (0.84±0.07 and 1.45±0.23) (P<0.01).Compared with the negative and blank control groups, the ERCC1-shRNA1 group showed a significantly decreased IC50 value (16.71±2.33 and 16.69±1.69 vs 7.78±0.54, P<0.01) and an increased proportion of G0/G1 phase cells ([72.87±3.23] and [71.75±4.56] vs [82.99±4.23]%, P<0.05), with the cell cycle arrested in the G0/G1 phase.The apoptosis rate of the cells in the ERCC1-shRNA1 group was remarkably lower after treated with cisplatin at the concentrations of 6.25 and 12.5 μg/mL than at 0 μg/mL ([8.17±0.65] and [11.91±1.41] vs [29.97±3.14]%, P<0.05).Conclusion ERCC1-shRNA can inhibit the proliferation and enhance the apoptosis of A549/DDP cells by silencing the expression of the ERCC1 gene.

OBJECTIVE: To investigate the expression of vascular endothelial growth factor in the nasal mucosa of chronic rhinosinusitis with nasal polys patients, and explored the regulation of clarithromycin on VEGF. METHOD: RT-PCR was used to detect the expression of VEGF in nasal mucosa from healthy control and CRSwNP. Nasal mucosal tissue explant culture measure and ELISA were used to explore the effect of clarithromycin on VEGF expression. RESULT: (1) VEGF mRNA expression level was significantly increased in CRSwNP compared with control and showed a statistic difference (P < 0.01). (2) There was a significant decrease in CRSwNP group undergo clarithromycin treatment on protein expression level of VEGF and showed a statistic difference (P < 0.05). CONCLUSION VEGF were overexpressed in CRSwNP group, which presume that play an important role in the pathogenesis of nasal polyps. Clarithromycin may play a therapeutical role on chronic rhinosinusitis through down-regulated the expression of VEGF.
Adult ; Chronic Disease ; Clarithromycin ; pharmacology ; Female ; Humans ; Male ; Nasal Mucosa ; metabolism ; Nasal Polyps ; complications ; Rhinitis ; complications ; metabolism ; Sinusitis ; complications ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

Objective To sum up the experience of position nursing of patients with ankylosing spondylitis and severe kyphosis. Methods Eleven patients with ankylosing spondylitis and severe kyphosis underwent surgery. The patients′spine and head were kept in a line during anesthesia to avoid spinal cord injury. The eye mask was used to make sure the eyelid close, so the bulbar conjunctiva and corneal did become dry. The angle of operation bed in time during operation was adjusted. The crushed area was observed and the position was changed to avoid pressure sores and edema. After surgery, they were keep inactive to make sure the spine stable and relieve the pressure. Results The 11 patients′surgery were successful. There were no pressure ulcer, fracture, spinal cord injury or eye injury occurs. The operation time was 9.3 ± 2.9 h. Conclusion Position nursing is very important to improve the success rate of surgery and reduce complications.

OBJECTIVETo investigate the protective effect of dexmedetomidine (Dex) preconditioning against ischemia/reperfusion (I/R) injuries in isolated rat hearts and its relation to mitochondrial permeability transition pore (mPTP) and mitochondrial ATP-sensitive K(+) channel (mitoKATP).

METHODSThe hearts of male SD rats were isolated to mount on the Langendorff apparatus and subjected to 30 min global ischemia followed by 120 min reperfusion. The isolated hearts were treated with Dex (10 nmol/L) before ischemia for 15 min. The left ventricular hemodynamic parameters,coronary flow (CF) and the lactate dehydrogenase (LDH) release in the coronary effluent at 5 min reperfusion were measured. The formazan content was assayed to determine the myocardial viability at the end of reperfusion.

RESULTSCompared with normal controls, I/R markedly decreased the left ventricular developed pressure and CF during the whole reperfusion period and the formazan content; while the left ventricular end diastolic pressure and LDH release were significantly increased. Dex preconditioning markedly improved the myocardial viability and cardiac function (P<0.01), which were reversed by the treatment with both atractyloside (20 μmol/L before ischemia), an opener of mPTP, and 5-hydroxydecanoate (100 μmol/L at the beginning of reperfusion), an inhibitor of mitoKATP, for 20 min.

CONCLUSIONDex has protective effect against I/R injuries in isolated rat hearts, which may be related to inhibiting the opening of mPTP at the beginning of reperfusion and activating mitoKATP before ischemia.

Animals ; Dexmedetomidine ; pharmacology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; Male ; Mitochondrial Membrane Transport Proteins ; drug effects ; Myocardial Reperfusion Injury ; physiopathology ; prevention & control ; Potassium Channels ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To explore whether hypoxia-inducible factor-1α( HIF-1α )is involved in oxygenglucose deprivation (OGD)induced caspase-independent cell death in primary cortical neurons and whether their expression is infected by necrostatin-1 ( Nec-1 ).Methods ( 1 ) Primary cerebrocortical neurons were cultured for 14 days.Pretred z-VAD.Fmk (z-VAD)and Nec-1 with 0.1,1,5,10,25 and 50 μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours,then cell viability was determined by measure LDH level.(2)Pretred z-VAD before the neurons were exposed to OGD for 2 hours,then reoxygenated for 0,2,6,12,24and 48 hours.Then western blot analysis protein level of HIF-1 α ;rt-PCR check its RNA level.(3)Pretred z-VAD and Nec-1 with 25μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours.Then western blot analysis protein level of HIF-1 α; rt-PCR check its RNA level.Result ( 1 ) When cells were pretread Nec-1 with 5 μ mol/L(6.97 ± 0.06),the level of LDH was lower than cells untreated( 14.23 ± 0.08 ) (P＜ 0.05);At 25 μmol/L( 2.21 ± 0.05),the level of LDH was essentially the same as that of the control( 1.03 ±0.03 ) (P＞0.05).(2)The protein level of HIF-1 αwas different from normal (0.24 ±0.01 ) when exposed to OGD for 2 hours and reoxygenated for 2 hours (0.57 ± 0.09) and was highest after cells were exposed to OGD for 2 hours and reoxygenated for 12 hours(0.91 ± 0.08 ) (P＜ 0.05 ).The RNA level of HIF-1 α when cells were exposed to OGD was not deferent from normal (P ＞ 0.05 ).( 3 ) When cells were pretread with Nec-1 (0.32 ± 0.04 ),the protein level of HIF-1α were lower than untreated(0.83 ±0.03) (P＜0.05),but the RNA level of HIF-1α had no deference(P ＞ 0.05).Conclusion HIF-1α was involved in cell' s caspase-independent cell death;Nec-1 can protect neurons through inhibiting the expression of HIF-1α.

Objective To evaluate the efficacy of caudal block with dexmedetomidine mixed with lidocaine for management of perioperative analgesia in children. Methods Thirty pediatric patients, aged 2-6 yr, weighing 8-23 kg, scheduled for elective unilateral high ligation of hernial sac, were equally and randomly assigned into either lidocaine group ( group L ) or dexmedetomidine mixed with lidocaine group ( group DL) using a random number table. Each patient received a single caudal dose of 1% lidocaine 1 ml∕kg in group L. Each patient received a single caudal dose of 1% lidocaine 1 ml∕kg mixed with dexmedetomidine 1 μg∕kg in group DL. Postoperative analgesia was assessed using FLACC scale. When FLACC score ≥4, ibuprofen suspension 10 mg∕kg was given orally. The consumption of ibuprofen was recorded within 8 h after operation. The onset time of caudal block and duration of analgesia were recorded, and adverse effects were observed. Results Compared with group L, the onset time of caudal block was significantly shortened, the duration of analgesia was prolonged, and the requirement for ibuprofen was decreased in group L. There was no significant difference in adverse effects between the two groups. Conclusion Addition of dexmedetomidine 1 μg∕kg to caudal lidocaine can significantly optimize the efficacy of caudal block with lidocaine alone for the management of perioperative analgesia in children.