Survey of vitamin D status in apparently healthy younger and elder adults / 中华检验医学杂志

Objective To analyze the vitamin D status among apparently healthy younger and elder adults in Beijing based on liquid chromatography tandem mass spectrometry.Methods This is an observational study.Participants included 287 apparently healthy young adults(143 males and 144 females) with an average of (32.2 ± 6.9) years old (19-44 years).At the same time 198 middle-and elder-aged adults were recruited [90 males,108 females,(55.6 ± 7.6) years],and fasting blood samples were collected and serum were isolated.They measured 25-hydroxyvitamin D (25OHD:25OHD2 and 25OHD3)using liquid chromatography tandem mass spectrometry method.Vitamin D with deficiency,insufficiency,sufficiency and intoxication was categorized as 25OHD ＜ 20 ng/ml,20-30 ng/ml,30-150 ng/ml,and ≥ 150 ng/ml,respectively.ALT,Ca,P,Cr,Glu,TG,TC and iPTH wereanalyzed using automatic analyzers.Statistical analysis was performed using SPSS17.0.Results The median 25OHD level in the total studied younger adults was 16.0 [2.5％-97.5％:(6.1-29.0) ng/ml] which didn't show significant difference with that of middle-and elder-aged adults.Younger males had significant higher level of 25OHD than females [17.9 (8.3-32.3) ng/ml vs.14.4 (5.4-26.4) ng/ml,Z =-4.238,P ＜ 0.01].Of the total younger subjects,the rate of vitamin D with deficiency (＜ 20 ng/ml),insufficiency (20-30 ng/ml)and sufficiency (≥30 ng/ml) was 72.8％,25.1％,2.1％,respectively,while that of middle-and elderaged adults was 76.3％,21.2％,2.5％ respectively,and that of younger males was 65.0％,30.8％,4.2％,respectively while that of younger females was 80.6％,19.4％,0％,respectively.Younger females had significantly higher rate of 25OHD deficiency (x2 =31.766,P ＜ 0.01).With adjusting sex,age and BMI,serum iPTH (r =-0.264,P ＜ 0.01) was significantly negatively correlated with 25OHD while Cr (r =0.221,P ＜ 0.01) showed significantly positively correlation with 25OHD.Conclusion Vitamin D deficiency is prevalent in both younger and elder adults in Beijing,especially in younger females.

Quantification of human urine and serum iodine by inductively coupled plasma mass spectrometry / 中华检验医学杂志

Objective This paper aims at establishing a inductively coupled plasma mass spectrometry ( ICP-MS) method for quantification and evaluation of iodine in human urine and serum in routine clinical laboratory .Methods This study was methodology validation research on iodine evaluation using ICP-MS.Ammonia, isopropanol and ultrapure water were mixed at certain ratio to dilute samples in the ratio of 1:10, and then the diluted samples were analyzed by ICP -MS.Re was used as the internal standard.And linearity, lower limit of detection, recovery, precision, accuracy, carryover and stability was evaluated thoroughly .Results of iodine of pregnant women who required iodine tests were retrospectively analyzed to evaluate the status of iodine .Results The method only needs 30s for analysis of one sample .It was sensitive with a lower limit detection of 0.87μg/L, the correlation coefficient was higher than 0.999 9 in ten measurements.The recovery in both serum and urine was approximately 100% (95.3% -109.9%). Based on the NIST standard reference material 3668 comparison, the bias was less than 4%( -0.9% -3.9%).The inter-coefficient variation (CV) for serum iodine and urine iodine was 1.2%-3.0%, 2. 0%-2.9%, respectively;and total CV for serum iodine and urine iodine were 3.0%-3.8%, 4.1%-4.9%, respectively.The mean carryover of this method was 0.03% and iodine was stable for at least one month at -20℃ and 4℃.The urine and serum iodine for pregnant women was (154.8 ±89.7) μg/L (mean ±SD),(75.8 ±21.4) μg/L, respectively.The correlation between urine and serum iodine was 0.21. Conclusion Establishe a rapid and simple ICP -MS method for urine and serum iodine measurement with high accurate and precise in routine clinical laboratory .

Molecular diagnosis of thalassemia compound heterozygotes in PUMCH from 2012 to 2015 / 中华检验医学杂志

Objective To investigate the genotype distribution of thalassemia intermedia , major and compound thalassemia in Peking Union Medical College Hospital from 2012 to 2015. Methods Retrospectively 1 084 suspected thalassemia cases were analyzed in recent four years .Three common deletions of αglobin chain were detected by GAP-PCR.Three common point mutations of αglobin chain and seventeen common mutations of βglobin chain were identified by PCR reverse dot blot hybridization . Hemoglobin electrophoresis was carried out by Capillary Electrophoresis System .RBC associated parameters and morphology were analyzed by hematology analyzer and blood smear .Results 702 cases were confirmed to be thalassemia, and the positive rate was 64.76% (702 /1084).19 types of gene defects were detected. There were 4 types of gene defects in 23 case with α-thalassemia intermeida, including -α3.7 /--SEA , -α4.2 /--SEA , αCSα/--SEA and αQSα/--SEA , -α3.7 /--SEA to be the most common genotype (18 cases) .3 cases with β-thalassemia intermeida were confirmed and the genotypes were βCD 17(A→T) /β-29(A→G) , β-28(A→G) /β-28(A→G) andβIVS-Ⅱ-654(C→T) /βCD17(A→T) , respectively.There were also 1 βCD 41 -42(-TTCT) /βCD17(A→T) thalassemia major case. The genotypes of 2 HbE/β-thalassemia cases were βCD41 -42(-TTCT) /βE and βCD17(A→T) /βE.5 αβ-thalassemia including 2 βCD 41 -42(-TTCT) /βA compounded with αα/-α3.7 , 1βIVS-Ⅱ-654(C→T) /βA compounded with --SEA /αCSα, 1βCD17(A→T) /βA compounded with -α4.2 /ααand 1βCD 41 -42(-TTCT) /βA compounded with αCS α/αα.Rare and untypical haematological results were found , such as normal level HbA 2 and undetectable HbH, in compound heterozygosity with --SEA /αCS α and βIVS-Ⅱ-654(C→T) /βA. Conclusions The genotypes of thalassemia intermedia, major and compound thalassemia in Peking Union Medical College were highly variable .

Effect of decorin on proliferation and collagen type I synthesis of stiff knee joint synovial type B cells / 中国病理生理杂志

AIM:To explore the effects of decorin on procollagen type I (PcI), mRNA expression,collagen type I synthesis and proliferation of synovial type B cells of stiff knee joint synovial membrane.METHODS:Type B cells of synovial membrane were isolated from the stiff knee joint synovial membrane and cultured in vitro.The cells were treated with decorin at concentrations of 0.1 mg/L, 5 mg/L and 10 mg/L.After cultured for 24 h, 48 h and 72 h, the cell proli-feration rates were measured by MTT colorimetric determination.Cell cycle distribution and apoptosis were analyzed by flow cytometry.The mRNA level of Pc I was detected by RT-PCR, while collagen type I was measured by Western blot.RE-SULTS:The proliferation of synovial type B cells was significantly inhibited, the percentage of synovial type B cells at G1 phase was significantly increased by 5 mg/L and 10 mg/L decorin (P<0.05), and PcⅠmRNA expression and collagen type I synthesis were significantly decreased.The cells with late apoptosis were not found in control group and experimental groups.CONCLUSION:Recombinant human decorin inhibits synovial type B cell proliferation and decreases PcⅠmRNA expression and collagen type I synthesis in synovial type B cells of stiff knee joint synovial membrane in vitro, suggesting that decorin potentially contributes to the therapy of human knee stiffness.

Comparison of five automated 25OHD immunoassays and liquid chromatography tandem mass spectrometry method / 中华检验医学杂志

Objective To compare the concordance of five automated 25OHD immunoassays with liquid chromatography tandem mass spectrometry method ( LC-MS/MS) .Methods During May to July in 2014, 245 clinical serum samples that requested 25OHD tests were selected, with a total 25OHD range of 2.8 ng/ml-64.0 ng/ml, in which 154 samples did not contain 25OHD2 and 91 samples contains both 25OHD2 and 25OHD3 .To used a LC-MS/MS method that built in our laboratory to measure 25OHD, five commercial automated chemiluminescent immunoassays from Abbott Diagnostics ( A ) , DiaSorin LIASON (B), IDS-iSYS(C), Roche Diagnostics(D), and Siemens ADVIA Centaur(E).Taking the reference method LC-MS/MS as a standard , to compared the concordance and performance of the five automated 25OHD immunoassays.And used the commonly accepted cutoffs for 25OHDdeficiency (<20 ng/ml), and insufficiency ( 20 -30 ng/ml ) , and sufficiency (≥30 ng/ml ) to compare the uniformity of different methods .Statistical analysiswere performed by MedCalc software , Passing & Bablok regression , Bland &Altaman plots and Box and whisker plots were performed to compare the differences of the methods .Results The medium ( range:2.5% -97.5%) 25OHD of the 245 serum samples of the six methods was 23.5 (5.8-44.2) ng/ml(LC-MS/MS),20.6 (7.1-43.5)ng/ml(A),19.0 (5.4-38.0) ng/mL (B),23.0 (10.0-38.1) ng/ml(C),20.1 (5.1 -46.0) ng/ml (D),31.3 (12.3 -71.1) ng/ml (E), respectively .Passing and Bablok regression showed that method B had the best correlation coefficient with LC-MS/MS (r=0.894), while methods A, C and D had relatively small bias compared withLC-MS/MS and method E had the large bias .If the serum samples did not contain 25OHD2 , all the five automated immunoassays correlated well with LC-MS/MS with a correlation coefficient higher than 0.84, and B has the best correlation with LC-MS/MS ( r=0.930 ) .While all the correlation coefficient between immunoassays and LC-MS/MS decreasedwhen analyzing the samplescontaining 25OHD2.Using the clinical cutoffs, A, B, C, D and E had a concordance of 68.6%, 64.9%, 67.8%, 70.6% and 51.8% compared with LC-MS/MS, respectively .Conclusions There are significant differences between different detection systems of 25OHD.All the immunoassays results were affected by the existence of 25OHD2 .The concordance of serum 25OHD resultswas poor between different methods , and it may be necessary to built exclusive cutoffs for different methods.

Anterior retropharyngeal approach for treatment of C2/3 fracture and dislocation / 中华创伤杂志

Objective To investigate the surgical techniques and clinical effects of anterior retropharyngeal approach in treatment of C2/3 fracture and dislocation.Methods Twelve patients with C2/3 fracture and dislocation treated via anterior retropharyngeal approach between November 2011 and April 2013 were included in the study.There were 7 males and 5 females aged from 19 to 65 years (mean,35 years).Primary pathologies included 7 patients with traumatic C3 fracture,2 with Hangman fracture and 3 with fracture and dislocation of the anteroinferior margin of C2 vertebrae.C2-C4 vertebrae were exposed using anterior retropharyngeal approach,followed by C2/3 discectomy or C3 corpectomy,decompression,interbody cage fusion or titanium mesh cage fusion,and anterior internal fixation.Results Exposure of lesion was sufficient for all patients and all operations were completed under direct vision,with mean operation time of 140 minutes and mean blood loss of 120 ml.One patient with reduced tone after operation gradually recovered in a week; one with dysphagia after operation recovered in 3 months; one with skin necrosis 7 days after operation was recovered by changing dressing; for the rest,there were no complications of incision hematoma,infection,or asphyxia.Ten patients were followed up for mean 15 months,which showed bony fusion in mean 6 months.At final follow-up,no implant loosening or displacement occurred.Conclusion Anterior retropharyngeal approach to C2/3 fracture and dislocation provides sufficient exposure of lesions,minor trauma,and less bleedings and complications,but as the local anatomy is complicated,there indeed exists a learning curve of the approach.

Texo-pulsed Dendritic Cells Inhibited the Growth of Osteosarcoma Cells by Stimulating CTL Response in Vitro / 医药导报

Objective To investigate the stimulation of exosome derived from osteosarcoma cells suppressing cytotoxic T cells and the inhibitory effect of active T cells for surpressing osteosarcoma cells. Methods Exosome derived from tumor cells was isolated and purified by ultracentrifugation. Its morphology was observed with transmission electron microscope, and the major histocompatibility complex-I ( MHC-I) molecules were analyzed by western blot. Mice bone marrow-derived dendritic cells were pulsed with exosome. Surface membrane MHC-I molecules were analyzed with flow cytometry. The effect of active T cells on the growth of osteosarcoma cells were detected by MTT assay after the T cells being stimulated by exosome-pulsed dendritic cells. Results The exosome was round or near round corpuscle, and the diameter was about 50-100 nm by transmission electron microscope. The size was relatively homogeneous. Western blot showed that the exosome expressed MHC-Ⅰmolecules. Surface membrane CD80,MHC-I and II molecules were expressed on 77. 16%, 83. 21%, and 91. 26% of LPS-treated dendritic cells, respectively, which were up-regulated compared to untreated cells. Dendritic cells pulsed with exosome derived from osteosarcoma cells caused significantly higher T cells stimulation and osteosarcoma cells inhibition as compared to un-pulsed dendritic cells. (P<0.05). Conclusion T cells can inhibit the growth of osteosarcoma cells after being stimulated by exosome-pulsed dendritic cells in vitro.

Correlation of ApoE gene polymorphism and serum uric acid in Ningxia Hui Autonomous Region / 中华检验医学杂志

Objective Toexplore the correlation of ApoE gene polymorphism and serum uric acid levels in Ningxia Hui Autonomous Region , China.Methods A case-control study.October 2011 to November 2011, five hundred twenty eight ( 296 male, 232 female ) apparently healthy individuals were studied.Questionnaires and physical examinations were performed by standard operation procedure.Fasting blood was collected for biochemistry testing including serum lipid parameters , uric acid concentration and creatinine levels.The multi-ARMS PCR was applied to determine ApoE genotypes ,and the relation of ApoE genotypes with serum lipid parameters and uric acid levels were analyzed.Non-normal distribution were compared using cause and inspection.Results The common six kinds of ApoE genotype can be detected.The total cholesterol ( TC) ,low density lipoprotein cholesterol ( LDL-C) and uric acid ( UA) levels in different genotype subgroups had statistical differences.The individuals with ε2/3 genotype had a significantly greater reductions in TC and LDL-C levels but increment in uric acid concentration than those withε3/3 and ε3/4 genotype (P<0.05).The effect of ApoE gene polymorphism on uric acid levels still remained significantly after adjustment for age , gender , region and other factors.Conclusion The ApoE polymorphism is associated with serum uric acid levels and individuals with ε2 allele have higher serum uric acid levels.

Establishment and preliminary clinical application of serum bile acid spectrum analysis by ultra high / 中华检验医学杂志

ObjectiveTo establish an ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for detecting the bile acid expression in serum and to explore the relationship between the bile acid spectrum and the obesity.Methods After pre-treatment through solid phase extraction,serum samples were chromatographed by gradient elution on an UPLC HSS T3 column,and then mass spectrometric analysis of bile acid was performed by multiple reaction monitoring ( MRM ) mode with negative electrospray ionization (ESI).And its methodology performance, including specificity,linearity,sensitivity,imprecision and recovery,were verified according to the guideline of State Food and Drug Administration (SFDA).Furthermore,serum samples from 10 simple obesity subjects and 10 healthy controls were analyzed with this establishedmethod, and ranksum test was used for statistical analysis.Results This established UPLC-MS/MS method could simultaneously quantify 14 bile acid subgroups in serum sample,its analytical linear range was 10 - 20 000 nmol/L.For different bile acid subgroups,the limit of detection (LOD) was 0.02 - 7.90 nmol/L,limit of quantitation (LOQ) was 0.07 -44.20 nmol/L; and its within-day coefficient of variation (CV) was 0.35％ - 12.41％,between-day CV was 1.11％ - 13.04％ ; the relative recovery ratio of this method was 89.8％ - 114.6％.Some differenceswere observed in chromatogram between obesity and control subjects,and both free bile acid and conjugated bile acid concentrations in obesity subjects [ 0.49 ( 0.45 - 1.90 ),1.44 ( 0.84 - 3.72 ) μmol/L ] were lower than them in control subjects [0.98(0.53 -3.06),1.99(0.67 -2.88) μmol/L],but the difference was not significant ( Z =- 0.958,- 0.801,P ＞ 0.05 ).ConclusionsThis established UPLC-MS/MS method can efficiently differentiate and quantify 14 bile acid subgroups,and is characterized with broad analytical measurement range,good analytical sensitivity and precision.This method can be applied for the free and conjugated bile acid analysis in clinical specimens.

Post-marketed re-evaluation of fleabane injection and Dengzhan Shengmai capsule study on treatment in patients with ischemic stroke / 中国中药杂志

<p><b>OBJECTIVE</b>To verify the efficacy and safety of post-marketed fleabane injection combined with Dengzhan Shengmai capsules in the treatment of ischemic stroke (IS).</p><p><b>METHOD</b>A multicentre, prospective, practical, randomized controlled study was carried out to compare the efficacy and safety of Dengzhan group (n = 343) and western medicine group (n = 335), appling "clinical study central stochastic system". The treatment of Dengzhan group is using fleabane injection in acute stage and Dengzhan Shengmai capsules in convalescence. The primary indexes of effect evaluation are the important outcome events in 360 days' follow-up, including mortality, recurrence, disability and quality of life to reflect the effect of clinical study. The indexes of safety evaluation involve laboratory examination results and incidence of adverse events.</p><p><b>RESULT</b>After 360 days' follow-up, 4 people died of IS in Dengzhan group, and the mortality rate of which is 1.17%, while 16 died in Western medicine group (WM group), and the mortality rate is 4.78%, suggesting that the mortality rate of Dengzhan group is significantly lower than WM group (P<0.05). Eleven cases recurred in Dengzhan group, and the recurrence rate of which is 3.21%, while 12 recurred in WM group, and the recurrence rate is 3.59%, indicating that the recurrence rate of Dengzhan group is slightly lower than WM group. The disability rate of Dengzhan group is 39.53%, among which the rate of severely disabled cases are 1.49%, while the disability rate of WM group is 40.13%, among which the rate of severely disabled cases are 3.13%, suggesting that the disability rate of Dengzhan group is lower and the severity of disability is also lighter than WM group. In the field of quality of life, the activity ability and the upper limb function store of stroke patients in Dengzhan group improved far much better than WM group (P<0.05). Analysis of safety suggested that, adverse events occurred in 11 cases in Dengzhan group, among which 4 cases is related with the drug treatment, the incidence of adverse events of which is 1.17%, and the main manifestations involve fever and chilling, rash, nausea, dizziness, palpitation, etc. which were all appeared after the treatment of fleabane injection, and disappeared 1 to 2 days after drug withdrawal. 13 cases occurred abnormal liver function and 2 cases abnormal kidney function in Dengzhan group. According to the judgment of clinical physicians, 3 case of ALT abnormality is possibly related to the treatment, the others are all unrelated with the treatment.</p><p><b>CONCLUSION</b>Fleabane injection and Dengzhan Shengmai capsules are all safe and effective TCM in the treatment of ischemic stroke.</p>

Post-marketing re-evaluation of Kudiezi injection study on early treatment in patients with ischemic stroke / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect and safety of Kudiezi injection on patients with acute ischemic stroke.</p><p><b>METHOD</b>Seven hundreds patients were divided into two groups by central randomization system. The study group, 346 cases, was treated with kudiezi injection plus traditional Chinese medicine (TCM) synthesis rehabilitation project, and the control group, 354 cases, was treated with synthetic rehabilitation project. The patients were treated for 10 to 21 days. Before treatment and at the 7th, 14th and 21th day of treatment, the indexes include NIHSS used for evaluating the neurological deficit degree and the motor function score (Fugl-Meyer) for evaluating motor function were observed. The safety index is defined by adverse observation event and laboratory test. The incidence of adverse events and laboratory tests results were observed before and after treatment at the same time.</p><p><b>RESULT</b>Application of generalized estimating equation model, we found that as the treatment time, NIHSS score and FMI score of the two groups showed a trend of improvement. And at the 14th days and 21th days of treatment, compared to the control group the treatment group showed significant statistical difference on the impact of NIHSS and FMI (P<0.05). No serious adverse events were observed.</p><p><b>CONCLUSION</b>Kudiezi injection plus TCM rehabilitation project of ischemic stroke showed some superiority to western medicine rehabilitation program on improving the neurological deficit and motor function. Kudiezi injection is safe and effective in the treatment of acute ischemic stroke.</p>

Chronic tibial defection in children treated with grafting of fibula with vessel pedicles and Lebone under external fixators / 中华创伤骨科杂志

Objective To report one stage treatment of chronic tibial defection in children. Methods 19 children with chronic tibial defection were treated in our department when their infection was at the stage of latency. First their focuses of infection were cleared away. Next a piece of fibula with vessel pedicles whose length was equal to that of the tibial defect was taken as the support to restore the length of the tibia. Then the external fixators were applied and a mixture of antibiotic and Lebone powder was used to fill the defect. Results In 15 cases the incision healed at one stage, slight infection occurred in 4 cases whose incisions healed after treatment of 3 to 4 weeks. Follow-ups of 0.5 to 1.5 years showed that no infection occurred again, bone defects healed well and limb function was satisfactory. Conclusion To treat chronic tibial defection in children at one stage, combined therapy should be applied. Grafting of fibula with vessel pedicles and Lebone under external fixators is a good one, because it can shorten the treatment time at no cost of satisfactory effects.