Objective To observe the clinical efficacy of needle warming therapy plus pelvic floor muscle training in treating post-stroke neurogenic bladder.Method A total of 124 patients with post-stroke neurogenic bladder were divided into a treatment group and a control group by random number table, 62 cases in each group. The treatment group was intervened by acupuncture, moxibustion plus pelvic floor muscle training; the control group was intervened by Chinese patent drugs. The intervention was conducted with 2 weeks as a treatment course, for 3 treatment courses in total. The daily voiding frequency test and maximum cystometric capacity (MCC) in the two groups were measured before and after the intervention, and the clinical efficacies were compared between the two groups.Result Each item of the daily voiding frequency test in the two groups was significantly changed after the treatment (P<0.01). Each item of the daily voiding frequency test in the treatment group was significantly different from that in the control group after the treatment (P<0.01). The MCC was significantly changed after the intervention in both groups (P<0.01). The MCC in the treatment group was significantly different from that in the control group after the treatment (P<0.01). The recovery rate and total effective rate were respectively 35.5% and 96.8% in the treatment group versus 19.4% and 77.4% in the control group. There were significant differences in comparing the total effective rate (P<0.01) and the recovery rate between the two groups (P<0.05).Conclusion Needle warming therapy plus pelvic floor muscletraining can produce a significant therapeutic efficacy in treating post-stroke neurogenic bladder, and the therapeutic efficacy is obviously better than that of medication.

A novel unsaturated polyphosphoester (UPPE) was devised in our previous research, which is a kind of promising scaffold for improving bone regeneration. However, the polymerization process of UPPE scaffolds was unfavorable, which may adversely affect the bioactivity of osteoinductive molecules added if necessary, such as recombinant human bone morphogenetic protein-2 (rhBMP2). The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2 (UPB) and to investigate the bioactivity of rhBMP2 in this scaffold. Furthermore, the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro. A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres, and then the microspheres were added to UPPE for synthesizing UPB scaffold. The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy. The cumulative release of UPB scaffolds was detected by using ELISA. The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells (bMSCs) seeded on the surface of UPB scaffolds. The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates (ALP) activity in bMSCs seeded. The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2. The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h. bMSCs attached and grew on the surface of soaked UPB scaffolds, exerting well biocompatibility. The ALP activity of bMSCs seeded was significantly enhanced, indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds. It was concluded that UPB scaffolds have low cytotoxicity, good biocompatibility and preserve bioactivity of rhBMP2. UPB scaffolds are promising in improving bone regeneration.

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.

A novel unsaturated polyphosphoester (UPPE) was devised in our previous research, which is a kind of promising scaffold for improving bone regeneration. However, the polymerization process of UPPE scaffolds was unfavorable, which may adversely affect the bioactivity of osteoinductive molecules added if necessary, such as recombinant human bone morphogenetic protein-2 (rhBMP2). The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2 (UPB) and to investigate the bioactivity of rhBMP2 in this scaffold. Furthermore, the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro. A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres, and then the microspheres were added to UPPE for synthesizing UPB scaffold. The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy. The cumulative release of UPB scaffolds was detected by using ELISA. The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells (bMSCs) seeded on the surface of UPB scaffolds. The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates (ALP) activity in bMSCs seeded. The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2. The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h. bMSCs attached and grew on the surface of soaked UPB scaffolds, exerting well biocompatibility. The ALP activity of bMSCs seeded was significantly enhanced, indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds. It was concluded that UPB scaffolds have low cytotoxicity, good biocompatibility and preserve bioactivity of rhBMP2. UPB scaffolds are promising in improving bone regeneration.
Bone Morphogenetic Protein 2 ; chemistry ; pharmacology ; Bone Regeneration ; drug effects ; Humans ; Lactic Acid ; chemistry ; pharmacology ; Phosphatidylinositol Phosphates ; chemistry ; pharmacology ; Polyglycolic Acid ; chemistry ; pharmacology ; Tissue Scaffolds

Objective To assess the interference by calcium dobesilatein 7 peroxidase-baseduric acid assays and to determine its clinical significance.Methods In the in vitro experiments, uric acid in pooled serum with final concentrations of calcium dobesilate additions (0, 2, 4, 8, 16, 32, and 64μg/ml) were measured by 7 peroxidase-based assays.Percent Bias (%) was calculated relative to the drug-free specimen.In the in vivo experiments, changes in serum uric acid and calcium dobesilate concentrations were observed before and after calcium dobesilate administration ( baseline, 0 h, 1 h, 2 h, 3 h, 4 h, 6 h ) involunteers.The interference in different assays was assessed compared with LC-IDMS/MS method. Calcium dobesilate levels in 40 specimens from those taking calcium dobesilate were measured by HPLC method.Of the 40 specimens, 10 were selected to analyse the levels of uric acid by both peroxidase and UV measurement method to assess the impact in clinical status.Results In the in vitro study, concentrations of uric acid measured by 7 peroxidase-based assays were reduced by -6.3%to -21.2%compared with drug-free serum, when theconcentration of calcium dobesilate was16μg/ml.In the in vivo study, comparedto UA levels at 0 h, the biasesof serum uric acid determined by peroxidase method after calcium dobesilate administration(1 h, 2 h, 3 h, 4 h, 6 h) were of -3.33%, -6.79%, -7.49%, -6.07%, -4.09%, respectively.The observed uric acid concentrations for 8 participants measured by enzymatic assays were inhibited by -3.75% to -6.89% at 0 hour and by -16.9% to-22.22% at 2 hours relative to the concentrations measured by the LC-IDMS/MS method. Conclusions Calcium dobesilate produced a clinically significant negative interference with uric acid in all peroxidase-based uric acid assays,which may result in false evaluation of uric acid level in clinical status.Significant differences in the degree of interference were observed among the assays.

Objective To investigate the changes of prevalence of hyperuricemia ( HUA) and its correlations with blood glucose and lipid in healthy adults receiving physical examination at Peking Union Medical College Hospital (PUMCH) from 2012 to 2017. Meth-ods An observational approach was adopted for the data analysis.The test results of uric acid (UA),fasting blood glucose (FBG),to-tal cholesterol (TC),triacylglycerol (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C), creatinine (Cr) and Urea of 399 089 cases (206 881 males and 192 208 females) at PUMCH from January 2012 to December 2017 were collected and statistically analyzed.Results The total prevalence of HUA was 17.4% in which the prevalence of males was signif-icantly higher than that of females (25.6% vs 8.5%,χ2＝20 234.850,P＜0.01).During the years of 2012 to 2017,the prevalence of HUA was 26.5%,24.7%,28.6%,23.9%,24.8% and 24.5% in males,and 13.8%,6.3%,7.9%,6.1%,6.2% and 6.8% in females for each year respectively.The prevalence of HUA in males aged 18 to 64 years old was significantly higher than that in the age-matched fe-males (all P＜0.05).However, the prevalence of HUA in males aged≥65 years old was similar to the age-matched females.There was no statistically significant difference of HUA prevalence between males and females aged ≥65 in 2013,2015,2016 and 2017 ( χ2＝1.792,0.017,1.440 and 0.205 respectively;all P＞0.05).The percentages of hyperlipidemia in both males and females of HUA group were higher than those of non-HUA group respectively (all P＜0.01).The percentage of hyperglycemia in males of non-HUA group was higher than that of HUA group,but the percentage of hyperglycemia in females of non-HUA group was lower than that of HUA group ( all P＜0.01).High levels of TC,TG and FBG were risk factors of HUA with increased OR values in increased concentrations of TC,TG and FBG,respectively.Conclusion During the recent 6 years, in healthy adults receiving physiced examination at PVMCH, the preva-lence of male HUA diagnosed was at overall high level,but the prevalence of female HUA was in decreasing and relatively stable trend. Hyperlipidemia and hyperglycemia should be the risk factors of HUA.

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.

Objective:By exploring a construction and improvement system for total lab automation in emergency lab, this work aims to improve the success rates of project implementation and the effective utilization of medical resources.Methods:Firstly, clarify the short board of current solution and determine the configuration of the TLA solution which include the number of pre-analytic, post-analytic modules and analyzers by workflow and data analysis which based on the data from 16th to 22nd September 2019. Secondly, define the basic principles and design the optional solutions. Thirdly, determine the final solution based on the simulation results. Fourthly, evaluate the effectiveness of the solution by comparing the turnaroud time (TAT) before and after implementation.Result:We upgrade the system to improve the automation level of the track by add 1 online centrifuge module and upgrade the capacity of online storage module from 9 000 samples to 15 000 samples, and increase the throughput of the clinical chemistry analyzers form 2 500 tests/h to 5 400 tests/h and immunoassay analyzers from 668 tests/h to 1 320 tests/h with no more space. The 95th percentile TAT (from sample check in to validation) have 33% reduction on the premise of 9% increase on the workload of clinical chemistry and immunoassay, and it is highly consistent with the results obtained by data simulation whose 95th percentile TAT is 67 min.Conclusion:Good uasge of workflow and data analysis in the stage of solution design can help to ensure that the result can meet the expectations of the clinicians and effective utilization of medical resources.

Objective:To explore the mechanism of acupuncture combined with sand therapy on bone biomechanics and immune function in rabbits with knee osteoarthritis based on G protein coupled signal.Methods:Totally 35 rabbits were randomly divided into normal group, model group, sand therapy group, acupuncture and moxibustion group and combined group, with 7 rabbits in each group. Except for the normal group, all other groups were prepared with knee osteoarthritis models. After 12 h modeling, rabbits in the sand therapy group and the combined group were treated with sand therapy. Rabbits in the acupuncture and moxibustion group and the combined group received acupuncture stimulation intervention therapy. Rabbits in normal group and model group only moved freely. The levels of IgG, IgM and IgA in serum and IL-1β and TNF-α were detected by ELISA method. The biomechanical properties of femur were measured by American Instron universal material experimental machine. The pathological changes of rabbits in each group were observed by HE staining. Protein expression levels of Gαs, cAMP and Gi in rabbit cartilage tissues were detected by Western blot.Results:Compared with model group, the levels of IgG, IgM and IgA increased ( P<0.05)l the levels of IL-1β and TNF-α decreased ( P<0.05); the maximum load, breaking load, structural stiffness and capacity absorption of rabbits in sand therapy group, acupuncture group and combined group increased ( P<0.05); the expressions of Gαs and cAMP protein increased ( P<0.05), while the expression of Gi protein decreased ( P<0.05). Conclusion:Acupuncture combined with sand therapy in the treatment of knee osteoarthritis can effectively improve the bone strength of osteoarthritis, reduce the inflammatory response, balance the expression of G protein and improve the imbalance of immunoglobulin.

Objective:To investigate the effects of external application of Compound Dahuang Powder combined with acupuncture on the synovial ultrastructure, pathological angiogenesis, and Ras/Raf signaling pathway in rheumatoid arthritis rats.Methods:Totally 100 SPF grade SD rats were selected and divided into normal group, model group, acupuncture group, external application group, and combination group according to random number table method, with 20 rats in each group. Except for the normal group, Freund's complete adjuvant was injected into the bilateral plantars to establish a rheumatoid arthritis model. One week after modeling, the acupuncture group received acupuncture at "Yinlingquan (SP9)" and "Sanyinjiao (SP6)" for 30 minutes, with continuous intervention for 6 days and 7 days as one course of treatment, for a total of 3 courses of intervention. The external application group of rats applied compound Dahuang Powder on the right hind foot and ankle joint, 10 g/time, twice a day, for 7 days as one course of treatment, with a total of 3 intervention courses; the combination group received a combined intervention of acupuncture and external application of compound Dahuang Powder. No intervention was performed in the normal group and model group. The levels of TNF-α and IL-1β in serum of rats in each group were detected using enzyme-linked immunosorbent assay. Transmission electron microscopy was used to observe the ultrastructure of synovial cells. HE staining was used to observe the morphology of synovial tissue. Immunohistochemistry was used to detect vascular endothelial growth factor (VEGF) and its receptor VEGFR2 in synovial tissue. Western blot and RT qPCR were used to detect Ras/Raf protein and mRNA in synovial tissue, respectively.Results:Compared with model group, the levels of TNF-α and IL-1β in serum of rats in acupuncture group and external application group decreased ( P<0.05), the expressions of VEGF and VEGFR2 decreased ( P<0.05), and the expressions of Ras and Raf protein and mRNA in acupuncture group and external application group decreased ( P<0.05). Compared with acupuncture group and external application group, the levels of TNF-α and IL-1β in serum of rats in the combination group decreased ( P<0.05), the expressions of VEGF and VEGFR2 decreased ( P<0.05), and the expressions of Ras and Raf protein and mRNA in the combination group decreased ( P<0.05). The results of synovial cell ultrastructure showed that the nuclear membrane of synovial cells in normal group was good, and the nuclear membrane in model group was seriously damaged. The acupuncture group and external application group had different degrees of improvement, and the combination group had the most significant improvement. HE staining results showed that the joint synovial tissue of the normal group was normal, the joint synovial tissue of the model group had a large amount of inflammatory infiltration, the joint synovial injury of the acupuncture group and the external application group was slightly improved, and the pathological injury of the combined group was significantly reduced. Conclusion:The combination of external application of compound Dahuang Powder and acupuncture can regulate angiogenesis related factors, regulate the Ras/Raf signaling pathway, and improve the ultrastructure of synovial cells in rheumatoid arthritis model rats.