The preservation of urinary proteins on a membrane plays a vital role in biomarker research, and the efficient elution of proteins preserved on nitrocellulose membrane (NC membrane) determines the application of this method. During the heating elution procedure, we raised the temperature to reduce the intense vortexing time, and kept gentle rotating while precipitation to prevent nitrocellulose reformation. We also used SDS-PAGE and LC-MS/MS to analyze the urinary proteins prepared by heating elution procedure, intense vortexing elution procedure and acetone precipitation method. There was no degradation of proteins prepared by heating elution procedure. Compared with proteins prepared by heating elution method and acetone precipitation method, the overlapping rates of the proteins was almost the same (92.6% versus 96.8%) and the ratios of CV values (< 20%) of the proteins were both high (85.2% and 94.4%). The heating elution procedure achieved good technical reproducibility, and was much simpler and more efficient than the previous one. It can facilitate the application of the preservation of urinary proteins on membrane.
Acetone ; Biomarkers ; urine ; Chromatography, Liquid ; Collodion ; Electrophoresis, Polyacrylamide Gel ; Hot Temperature ; Humans ; Proteins ; isolation & purification ; Reproducibility of Results ; Tandem Mass Spectrometry ; Urine ; chemistry

BACKGROUND: International research discovers that the risk factors of stroke have certain correlation with microalbuminuria(MAU) but the results are inconsistent.OBJECTIVE: To investigate the relationship between the incidence of MAU in patients with acute cerebral infarction and the risk factor of stroke and the recurrence of cerebral infarction.DESIGN: A case-control study based on patients with acute cerebral infarction and individuals with risk factors of cerebral infarction, and the healthy elderly as controls.SETTING: Inpatient department of neurology in a university hospital.PARTICIPANTS: Totally 214 cases with cerebral infarction or risk factors of cerebral infarction aged between 50 and 80 years old were selected from the Department of Neurology of the First Affiliated Hospital of Chongqing Medical University between January 2000 and December 2001. Acute cerebral infarction group (infarction group, attack within one week, n =78), cerebral infarction risk factor group(risk group, n = 56), such as, hypertension, diabetes,coronary heart disease (CHD), transient ischemic attack(TIA) or stroke history(more than 6 months), and healthy elderly(control group, n=28) .Thereinto, combined group( n = 109) included 78 cases of acute cerebral infarction and 31 cases with TIA or stroke history of risk factors.METHODS: Seralbumin, serum creatine(SCr), blood sugar, total cholesterol, total triacylglycerol(TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol(LDL-C) were detected after more than 12 hours of fasting. The albumin in the first urine in the morning was detected by radio-immunity analysis.the incidence of neo-vascular affair in 2 years of follow-up including recurtute indicator: incidence of MAU in each group.RESULTS: Patients with MAU accounted for 35% of acute cerebral infarction, which were significantly higher than that of risk group(15% ) and control group(3.5%, P ＜ 0.01). Six independent risk factors predicting MAU existed in infarction group and combined group, which were diabetes, seralbumin level, age, CHD, hypertension, and HDL-C( P ＜ 0. 05, or P ＜ 0.01).After(1.5 ±0. 9) years of follow-up, 23% of patients of infarction group suffered from recurrent vascular accident, which was significantly higher than that of risk group(12% ) and control group(0%, P ＜ 0.01 ). 36% of MAU patients in infarction group suffered from recurrent vascular accident, which was significantly different from that of patients with a great deal of albuminuria(41% ) and normal albuminuria(10%, P ＜ 0. 05). MAU is the independent predicting indicator for the recurrence of stroke after the control of diabetes and hypertension in patients of infarction group and risk group with the risk rate of 3.8 (95% CI, 1.2-13.5; P ＜0.05) or 4. 7(95%CI,1.4-16.7, P ＜ 0.01).CONCLUSION: MAU is quite common in acute cerebral infarction patients. After the adjustment of basic risk factors, MAU is the independent risk factor of the recurrence of cerebral infarction.

Pericytes are a very important cellular constituent of the blood-brain barrier.They play a regulatory role in brain angiogenesis,endothelial cell tight junction formation,blood-brain barrier differentiation,microvascular dynamic motion and structural stability.Pericytes exhibit unique functional characteristics in some diseases,such as cerebrovascular disease,neurodegenerative disease,neuroimmune disease and traumatic brain injury.This article reviews the roles of pericytes in the blood-brain barrier.

Objective To analyze the changes in the composition of hospitalization of thyroid disease in Lianyungang City from 2002 to 2010,to learn the impact of iodine intake on thyroid disease in Lianyungang City after implementing universal salt iodization (USI),and to get epidemiologic evidence for prevention and control of iodine deficiency disorders scientifically.Methods Medical reports of the patients with thyroid disease were collected from hospitals at county level and above institutions in Lianyungang City from 2002 to 2010.Monitoring results of household iodized salt were also collected from five counties (districts) in Lianyungang City.The changes in the composition of hospitalization of thyroid disease were analyzed,and the relationship between thyroid disease and iodized salt was studied.Results ① A total of 3 559 patients with thyroid disease were enrolled in the study,the ratio between male and female was 1 ∶ 3.36.Mean age was (46.81 ± 14.94) years old and the peak age was from 40 to 60 years old.② Among the hospitalized patients with thyroid disease,the proportion of goiter patients,hyperthyroidism patients,thyroiditis patients,hypothyroidism and other thyroid disorders was 41.64％ (1 482/3 559),40.07％ (1 426/3 559),6.83％ (243/3 559),6.21％ (221/3 559) and 5.25％ (187/3 559),respectively.③In 2002-2010,the hospitalization rate of thyroid disease showed an upward trend (x2 =460.12,P＜ 0.01),and the hospitalization rates of hypothyroidism,goiter,hyperthyroidism,thyroiditis and other thyroid disorders were all increased year by year (x2 =50.54,281.91,85.15,19.51 and 65.01,all P ＜ 0.01).④From 2002 to 2006,among the patients with thyroid disease,the proportion of hyperthyroidism was the highest,and the range of values was from 42.03％ (124/295) to 53.02％ (114/215).Followed by goiter,and the range of values was from 34.55％ (142/411) to 39.25％ (73/186).From 2007 to 2010,the proportion of goiter became the highest,and the range of values was from 42.11％ (232/551) to 49.57％ (291/587).Followed by hyperthyroidism,and the range of values was from 33.05％ (194/587) to 40.47％ (223/551).⑤From 2002 to 2010,the coverage of iodized salt and the consumption rate of qualified iodized salt both increased year by year (x2 =183.75,211.99,all P ＜ 0.01).⑥There were positive correlations between the hospitalization rates of thyroid disease,hypothyroidism,goiter,other thyroid disorders and the coverage of iodized salt,and the consumption rate of qualified iodized salt (rs =0.83,0.93,0.82,0.87,0.83,0.93,0.74 and 0.91,all P ＜ 0.05).⑦There was a positive correlation between the hospitalization rate of hyperthyroidism and the consumption rate of qualified iodized salt (rs =0.72,P ＜ 0.05).Conclusions There are some changes in the composition of hospitalization of thyroid disease in Lianyungang city from 2002 to 2010.Iodine intake may affect the hospitalization rate of thyroid disease.

Objective To investigate and analyze the personality characteristics and situation of mental health in patients with sudden deafness,and discuss the relationship between the two,so as to provide basis for the auxiliary diagnosis and mental health intervention.Methods Eighty-eight patients with sudden deafness(sudden deafness group) were evaluated by Symptom Checklist 90 (SCL-90) and adult Eysenck Personality Questionnaire (EPQ).The survey results were compared with the norm of healthy people (norm group).The correlation analysis was performed between SCL-90 results and EPQ score.Results The SCL-90 total score in sudden deafness group was higher than that in normal group,but there was no significant difference,P>0.05.The scores of somatization,depression,anxiety,panic,psychosis and positive number,positive projects split were (1.58±0.46),(1.71±0.43),(1.59±0.44),(1.38±0.39),(1.46±0.51) scores,(32.65±15.81) nubmers,(2.96±0.65) scores in sudden deafness group,which were higher than those in normal group (1.37±0.48),(1.50±0.59),(1.39±0.43),(1.23±0.41),(1.29±0.42) scores,(24.92±18.14) nubmers,(2.60±0.59) scores,and there were significant differences,t=3.640,2.996,3.855,3.045,3.314,3.562,5.033,P＜0.01.The standard T points of psychotic,neurotic and emotional stability dimensions were (54.63±10.37),(45.59±8.67) scores in sudden deafness group,which were higher than those in normal group (48.14±9.74),(41.27±9.42) scores,and there were significant differences,t=8.207,3.811,P＜0.01.The standard T points of internal and external dimensions,hide were (52.28±11.67),(48.68±8.57) scores in sudden deafness group,which were lower than those in normal group (57.75±8.26),(51.27±10.43) scores,and there were significant differences,t=5.455,2.066,P＜0.01 or 0.05.Neurotic and emotional stability dimension,psychotic part dimension were a positive correlation with SCL-90 factor points.Multi-factor stepwise regression analysis showed that the neurotic or emotional stability were correlated with interpersonal relationship,depression,anxiety,hostility,terror.Conclusions Patients with sudden deafness overall mental health status is poorer,the somatization,depression,anxiety,terror,psychoticism of various factors on negative emotions is more obvious.Sudden deafness patients have emotional instability and psychotic personality characteristics.The personality characteristics of sudden deafness patients have a correlation with the mental health.In the process of comprehensive treatment for patients with sudden hearing loss,individual psychological health intervention is necessary.

AIM:To establish a HPLC method for determining contents of tetrahydropalmatine and pepper alkali in Zhitong Capsule(Rhizoma Corydalis,Fructus Piperis,Radix Paeoniae alba,etc.)to control its quality.METHODS:The samples were extracted in water by ultrasonic wave,and then extracted by aether to refine after being acidized by 1 mol/mL HCl.Following that the pH of water solution was adjusted to 9.0-10.0 by NH_3?H_2O,and then extracted by aether again.After that,the aether solution was collected to evaporation to dryness and metered by methanol.The sample solution was determined by high performance liquid chromatography on a Hypersil ODS 2 C_ 18 column(4.6 mm?250 mm,5 ?m)with mobile phase composed methanol-0.1% phosphoric acid water solution(the pH was adjusted to 6.0 by triethylamine)(55∶45).The flow rate was 1.0 mL/min,the column temperature was at 30 ℃ and the signal from 0 to 14.5 minutes were acquired at 280 nm,and that from 14.5 to 22 minutes were detected at 328 nm.RESULTS:The resolution of tetrahydropalmatine and pepper alkali was good,with no miscellaneous peak.The linear range was at 0.196-1.96 ?g(r=0.999 6)for tetrahydropalmatine,0.03-0.3 ?g(r=0.999 7)for pepper alkali.The average rate of recovery of tetrahydropalmatine was 97.74%(RSD= 0.42%,n=6),and that of pepper alkali was 104.51%(RSD=2.01%,n=6).CONCLUSION:The method is simple,reliable,accurate and can be applied to the quality control of the preparation.

Scientific research training program is an important component in the 8-year medical education program. It focuses on the training of scientific research thinking and research methods. It is necessary to establish an effective evaluation system in order to achieve the learning outcomes of scientific research training program. PUMC has established an effective evaluation system which involves research project selecting,supervising,formative assessment,oral defense. This paper introduces the evaluation system of scientific research training program at PUMC.

This is a review of student enrollment by nursing school of PUMC from 2003 to 2006.The paper covers the introduction of national policy for student enrollment,analysis of PUMC student background and challenge in the processing of student admission.The information supports the reformation of enrollment in terms of matriculation including objective and constructed interviewing approach.

Objective To study the effects and mechanisms of resveratrol (Resv) on the expression of adiponectin receptors ( AdipoR1 and AdipoR2 ) in rat mesangial cells (MCs) cultured in intermittent high glucose medium.Methods The MCs cultured in normal glucose ( 5.6 mmol/L) medium were transfected by the plasmid vector of the FoxO1 short hairpin RNAs ( FoxO1 shRNA ),and then cultured in intermittent high glucose medium (5.6mmol/L or 30 mmol/L,alternately every 8 hours ) and resveratrol ( Resv 20 μmol/L).The MCs cultured in normal glucose (5.6 mmol/L) medium served as normal glucose control group (NG),and the MCs exposed to iutermittent high glucose medium were divided into five groups:intermittent high glucose group (IHG),IHG+Resv group,IHG+FoxO1 shRNA group,IHG+ Resv + FoxO1 shRNA group,IHG + Negative control group ( shNC ),each group was cultured for 72 h.The level of reactive oxygen species (ROS) was assessed by Fluorescence microplate reader.The mRNA levels of Sirt1,Foxo1,AdipoR1,and AdipoR2 were assessed by RT-RCR.The protein levels of FoxO1,phosphorylation FoxO1 ( p-FoxO1 ),AdipoR1,and AdipoR2 were assessed by Western blotting.Results ( 1 )Compared with NG,intermittent high glucose significantly decreased the mRNA expression of Sirt1,but markedly increased the level of p-FoxO1.Furthermore,the mRNA and protein expression levels of AdipoR1 were obviously decreased while the level of ROS was enhanced in IHG ( all P＜0.05 ).(2) Compared with IHG,the mRNA and protein expression levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was enhanced in group IHG+FoxO1 shRNA ( all P＜0.05 ).( 3 ) Compared with IHG,after treating with Resv,the mRNA expression level of Sirtl was increased,whereas the level of p-FoxO1 was decreased.Moreover,the mRNA and protein levels of AdipoR1 was increased while the level of ROS was lowered in group IHG+Resv (all P＜0.05 ).(4) Compared with group IHG+Resv,the mRNA and protein levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was increased in group IHG+Resv+FoxO1 shRNA (all P＜O.05).(5) In addition,the mRNA and protein expression level of AdipoR2 showed no significant difference among these groups ( P＞0.05 ).Conclusion ( 1 ) Resveratrol significantly increases the expression of AdipoR1 in MCs induced by intermittent high glucose.( 2 ) FoxO1 plays an important role in regulating the expression of AdipoR1 by resveratrol.

BACKGROUND:Studies on high glucose exposure in human periodontal ligament cel s usual y focus on the biological behaviors, pathways and secretory factors, but whether the metabolic memory is involved is little known. OBJECTIVE:To investigate the metabolic memory of high glucose exposure in human periodontal ligament cel s. METHODS:Human periodontal ligament cel s were primarily cultured and identified. Cel s at 5-8 passages were selected and randomized into four groups. Group A (controls):DMEM containing 5.5 mmol/L glucose for 8 days;group B (5-day memory group):DMEM containing 35 mmol/L glucose for 3 days and DMEM containing 5.5 mmol/L glucose for 5 days;group C (3-day memory group):DMEM containing 35 mmol/L glucose for 5 days and DMEM containing 5.5 mmol/L glucose for 3 days;group D (8-day high glucose group):DMEM containing 35 mmol/L glucose for 8 days. The cel proliferation was detected by cel counting kit-8, the cel apoptosis was determined by flow cytometry, and the levels of total proteins and alkaline phosphatase were investigated using ELISA. RESULTS AND CONCLUSION:Compared with the control group, the cel proliferation in the other three groups was significantly reduced (P<0.05), the number of apoptotic cel s was significantly increased, while the levels of total proteins and alkaline phosphatase were significantly decreased (P<0.05). These results suggest that high glucose causes persistent changes in human periodontal ligament cel s by inhibiting cel viability, increasing the apoptosis and downregulating the levels of the total proteins and alkaline phosphatase