Objective To discuss the improvement for the verification of inter-day precision of PS activity assay and the verification protocol for limits of quantitative of FⅧ:C and FⅨ:C assay,aiming at the problems in the performance verification of anticoagulant protein S(PS),coagulation factor Ⅷ activity(F Ⅷ:C)and coagulation factor Ⅸ activity(F Ⅸ:C)assay.Methods SYSMEX CN-6000 and supporting reagents were used,and low-and high-value quality control(QCs)were used as the study samples.Following the American Clinical and Laboratory Standards Institute(CLSI)document EP15-A3,an inter-day precision verification study of the PS activity assay was performed with three reagent preparation methods designed(specification-required method,instrument's manual method and improved method).The specification-required method was carried out completely according to the specification,and the instrument manual method involved allowing the required reagents to stand in the device for 30 minutes based on the specification-required method,and the improved method mixed and dispensed the reagents needed to be based on the specification-required method.To study the bottle variation of the same batch of reagents for the same sample,the acceptable range of external quality assessment(EQA)of the National Center for Clinical Laboratories(NCCL)was used as the evaluation standard.Following the CLSI EP25 document,the PS activity assay reagent onboard stability verification was performed with the specification-required method and the improved method.Following the CLSI EP17-A2 document,WS/T 514-2017"Establishment and Verification of Detection Capability for Clinical Laboratory Measurement Procedures"and the International Committee for Standardization in Hematology(ICSH)guidelines,the LoQ for FVIII:C and FIX:C assays was verified.The verification results were passed if the requirements of the specification were met.Results The verification result of inter-day precision(CVWL:12.9%～21.6%)of PS activity assay according to the specification method and the instrumental manual method exceeded the requirements of the specification(<10%CVWL at high levels,<20%CVWL at low levels).The results of inter-day precision verification with the improved method(CVWL:2.9%and 4.5%)were consistent with the requirements of the specification.The bottle variation exceeded the acceptable range of EQA.The improved method corrected the reagent on-board stability verification results of reagents(relative deviations of-4.24%～9.97%)by the specifications(high levels<10%,low levels<20%).The LoQ verification results for FⅧ:C and FⅨ:C assay were by the product specifications(FⅧ:C:0.75%～1.46%and 0.74%～1.40%;FⅨ:C:0.71%～1.27%and 0.70%～1.32%).Conclusion An improved method to improve the inter-day precision of PS activity detection is provided,and LoQ verification protocol for F Ⅷ:C and FⅨ:C assay is provided for clinical laboratory reference.

Objective To discuss the improvement for the verification of inter-day precision of PS activity assay and the verification protocol for limits of quantitative of FⅧ:C and FⅨ:C assay,aiming at the problems in the performance verification of anticoagulant protein S(PS),coagulation factor Ⅷ activity(F Ⅷ:C)and coagulation factor Ⅸ activity(F Ⅸ:C)assay.Methods SYSMEX CN-6000 and supporting reagents were used,and low-and high-value quality control(QCs)were used as the study samples.Following the American Clinical and Laboratory Standards Institute(CLSI)document EP15-A3,an inter-day precision verification study of the PS activity assay was performed with three reagent preparation methods designed(specification-required method,instrument's manual method and improved method).The specification-required method was carried out completely according to the specification,and the instrument manual method involved allowing the required reagents to stand in the device for 30 minutes based on the specification-required method,and the improved method mixed and dispensed the reagents needed to be based on the specification-required method.To study the bottle variation of the same batch of reagents for the same sample,the acceptable range of external quality assessment(EQA)of the National Center for Clinical Laboratories(NCCL)was used as the evaluation standard.Following the CLSI EP25 document,the PS activity assay reagent onboard stability verification was performed with the specification-required method and the improved method.Following the CLSI EP17-A2 document,WS/T 514-2017"Establishment and Verification of Detection Capability for Clinical Laboratory Measurement Procedures"and the International Committee for Standardization in Hematology(ICSH)guidelines,the LoQ for FVIII:C and FIX:C assays was verified.The verification results were passed if the requirements of the specification were met.Results The verification result of inter-day precision(CVWL:12.9%～21.6%)of PS activity assay according to the specification method and the instrumental manual method exceeded the requirements of the specification(<10%CVWL at high levels,<20%CVWL at low levels).The results of inter-day precision verification with the improved method(CVWL:2.9%and 4.5%)were consistent with the requirements of the specification.The bottle variation exceeded the acceptable range of EQA.The improved method corrected the reagent on-board stability verification results of reagents(relative deviations of-4.24%～9.97%)by the specifications(high levels<10%,low levels<20%).The LoQ verification results for FⅧ:C and FⅨ:C assay were by the product specifications(FⅧ:C:0.75%～1.46%and 0.74%～1.40%;FⅨ:C:0.71%～1.27%and 0.70%～1.32%).Conclusion An improved method to improve the inter-day precision of PS activity detection is provided,and LoQ verification protocol for F Ⅷ:C and FⅨ:C assay is provided for clinical laboratory reference.

Objective:To analyze the operational efficiency and Total Factor Productivity (TFP) of township health centers in Ningxia from 2013 to 2022,in order to provide decision-making basis for optimize the resource allocation of township health centers in Ningxia. Methods:The BCC model of Data Envelopment Analysis (DEA) and Malmquist productivity index were used to analyze the operation efficiency and TFP of township health centers of Ningxia. Results:(1) From 2013 to 2022,the personnel input in Ningxia township health centers increased by 45.80％,the number of outpatient visits had no significant increase,and the number of inpatient visits decreased by 45.40％. (2) The average comprehensive technical efficiency of resource allocation was 0.658,the average of pure technical efficiency and scale efficiency were 0.747 and 0.880. (3) The average value of TFP index was greater than 1 in 7 counties,and less than 1 in the other 15 counties. Conclusion:The efficiency of township health centers in Ningxia was low. It is of great significance to improve the resource allocation and management ability of township health centers and strengthen the policy support for improving the operation efficiency of township health centers.

Objective:To analyze the operational efficiency and Total Factor Productivity (TFP) of township health centers in Ningxia from 2013 to 2022,in order to provide decision-making basis for optimize the resource allocation of township health centers in Ningxia. Methods:The BCC model of Data Envelopment Analysis (DEA) and Malmquist productivity index were used to analyze the operation efficiency and TFP of township health centers of Ningxia. Results:(1) From 2013 to 2022,the personnel input in Ningxia township health centers increased by 45.80％,the number of outpatient visits had no significant increase,and the number of inpatient visits decreased by 45.40％. (2) The average comprehensive technical efficiency of resource allocation was 0.658,the average of pure technical efficiency and scale efficiency were 0.747 and 0.880. (3) The average value of TFP index was greater than 1 in 7 counties,and less than 1 in the other 15 counties. Conclusion:The efficiency of township health centers in Ningxia was low. It is of great significance to improve the resource allocation and management ability of township health centers and strengthen the policy support for improving the operation efficiency of township health centers.

Objective: To compare the differences and similarities among the system of quality of life instruments for cancer patients (QLICP) V1.0, the quality of life questionnaire (QLQ) from European Organization for Research and Treatment of Cancer (EORTC) and Functional Assessment of Cancer Therapy (FACT) from Center on Outcomes, Research and Education (CORE) of America. Methods: Based on literatures and our measuring data from patients at hospitals, the constructs, characteristics and psychometrics of the systems above were analyzed and compared. Internal consistency reliability was assessed using Cronbach α coefficient for each domain, and test-retest reliability through calculating the Pearson correlation coefficient r between the first and second assessments as well as intra-class correlation (ICC). Construct validity was evaluated by Pearson correlation coefficient r (item-domains correlations) and factor analysis. The criterion-related validity was evaluated by correlating corresponding domains of two instruments. Responsiveness was assessed through comparing the mean difference between the pre-treatment and post-treatment with standardized response mean (SRM). Results: The instruments of three systems were of different outstanding characteristics with all psychometrics meeting requirements. Measurements for 12 types of cancers showed that the internal consistency reliability Cronbach α coefficient for the overall scale of QLICP (V1.0) was 0.67-0.92, and for FACT was 0.79-0.98. The test-retest reliability (r or ICC) for the overall scale of QLICP (V1.0) was 0.61-0.99, and for FACT was 0.60-0.98. The SRM for the overall scale of QLICP (V1.0) was 0.25-1.28, and for FACT was 0.11-0.83. However, the QLICP was of better construct (clear hierarchical structure with items→facets→domains→overall) and Chinese culture. Conclusion The instruments of three systems can be used as the instruments to assess quality of life for patients with cancer with selections basing on different settings.

As a dioxygenase, Ten-Eleven Translocation 2 (TET2) catalyzes subsequent steps of 5-methylcytosine (5mC) oxidation. TET2 plays a critical role in the self-renewal, proliferation, and differentiation of hematopoietic stem cells, but its impact on mature hematopoietic cells is not well-characterized. Here we show that Tet2 plays an essential role in osteoclastogenesis. Deletion of Tet2 impairs the differentiation of osteoclast precursor cells (macrophages) and their maturation into bone-resorbing osteoclasts in vitro. Furthermore, Tet2 mice exhibit mild osteopetrosis, accompanied by decreased number of osteoclasts in vivo. Tet2 loss in macrophages results in the altered expression of a set of genes implicated in osteoclast differentiation, such as Cebpa, Mafb, and Nfkbiz. Tet2 deletion also leads to a genome-wide alteration in the level of 5-hydroxymethylcytosine (5hmC) and altered expression of a specific subset of macrophage genes associated with osteoclast differentiation. Furthermore, Tet2 interacts with Runx1 and negatively modulates its transcriptional activity. Our studies demonstrate a novel molecular mechanism controlling osteoclast differentiation and function by Tet2, that is, through interactions with Runx1 and the maintenance of genomic 5hmC. Targeting Tet2 and its pathway could be a potential therapeutic strategy for the prevention and treatment of abnormal bone mass caused by the deregulation of osteoclast activities.
5-Methylcytosine ; analogs & derivatives ; chemistry ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; DNA-Binding Proteins ; physiology ; Genome ; Genomics ; Mice ; Mice, Knockout ; Osteoclasts ; cytology ; metabolism ; Proto-Oncogene Proteins ; physiology

Objective To investigate the inhibitory effect of 188 Re-labeled BaGdF5-poly ( ethylene glycol) ( PEG) nanoparticles ( NPs) on hepatoma cells, and explore the application of the radiolabeled NPs for SPECT imaging. Methods BaGdF5-PEG NPs were synthesized by hydrothermal method, and were fur-ther radiolabeled with 188Re using diethylene triamine pentaacetic acid (DTPA) as a coupling agent. The human hepatoma cells SMCC 7721 were treated with different concentrations of BaGdF5-PEG NPs, 188 ReO-4 or 188Re-DTPA-BaGdF5 NPs (14.8, 74.0, 370.0×104 Bq/ml) for 24 h, and then the cell proliferation rates were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. 188ReO-4 and 188 Re-DTPA-BaGdF5 NPs were administrated into normal rabbits via the ear vein, respectively. For the former, static SPECT/CT imaging were performed at 30, 60 min post-injection, and for the latter, dynamic SPECT images were captured within 10 min, and static SPECT/CT images at 30, 60, 120 min post-injec-tion. The rabbit VX2 tumor model was established, and a microcatheter was inserted into hepatic artery via the rabbit femoral artery, and then the mixture of 188 Re-DTPA-BaGdF5 NPs and lipiodol was injected into the tumor region. SPECT/CT imaging for VX2 tumor was performed at 30 min later. Data were analyzed by two-sample t test. Results The BaGdF5-PEG NPs were nearly square and the particle size was about 10 nm. The labeling yield of 188 Re-DTPA-BaGdF5 was 94.1％ at the optimum conditions. Moreover, it showed high stability in vitro and in vivo. In vitro, BaGdF5-PEG NPs did not exhibit obvious cytotoxicity even at a high concentration. Both 188 ReO-4 and 188 Re-DTPA-BaGdF5 could inhibit the proliferation of SMCC 7721 cells, but 188 Re-DTPA-BaGdF5 showed a significantly stronger inhibitory effect at the doses of 74.0 and 370.0×104 Bq/ml ( t values:4.21,4.09, both P<0.01) . In vivo, 188 ReO-4 was absorbed by maxillary glands and was quickly elimi-nated from blood via the kidneys. The 188 Re-DTPA-BaGdF5 NPs mainly accumulated in the liver and spleen. In addition, retention and accumulation of 188 Re-DTPA-BaGdF5 NPs in the liver tumor could be achieved by using transarterial intervention technique for drug delivery. Conclusion 188Re-DTPA-BaGdF5 NPs have cer-tain killing effects on hepatoma cells in vitro, and with the help of transarterial intervention technique, the NPs can be aggregated within liver tumor, where they not only can be used for SPECT imaging, but also have potential therapeutic effects.

BACKGROUND:Our previous studies have found that polygonatum sibiricum polysaccharide (PSP) promotes osteogenic differentiation of bone marrow mesenchymal stem cel s (BMSCs) by Wnt/β-catenin signaling pathway, but the molecular mechanism is unclear.OBJECTIVE:To investigate the effect of PSP promoting the osteogenic differentiation via Wnt signaling pathways in BMSCs after LRP5 silencing. METHODS:LRP5 interference vectors were constructed and then transfected into C57BL/6 mouse BMSCs cultured in vitro. The transfection efficiency of cel s was calculated under fluorescence inverted microscope and the expression of LRP5 protein was detected by western blot assay. The osteogenic potential of BMSCs after LRP5-siRNA transfection was analyzed by alkaline phosphatase staining, alizarin red staining and western blot assay. Effect of PSP on the osteogenic differentiation of LIRP5-silenced mouse BMSCs was detected by real-time PCR and dual luciferase assay. RESULTS AND CONCLUSION:Compared with the control group, the mineralization ability, the mRNA expressions of Runx2 and Osterix, and the protein expression of LRP5 were significantly decreased in the LRP5-siRNA group (P<0.05). PSP could promote LRP5-siRNA transfected mouse BMSCs differentiating into osteoblasts and significantly upregulated the expressions ofβ-catenin and Osterixin, and also induced the high expression of luciferase reporter gene (TOPFlash) containing wild type TCF binding sites (P<0.05). To conclude, LRP5 plays an important role in the process of mouse BMSCs differentiating into osteoblasts. PSP can promote the osteogenic differentiation of mouse BMSCs by activating the Wnt/β-catenin signaling pathway independent on LRP5.

Objective To investigate the correlativity between elasticity modulus and pathological severity in chronic pancreatitis (CP).Methods Twenty-one pigs were divided randomly into experimental group (n=18) and control group (n=3) using random number method.The main pancreatic duct (MPD) was incompletely ligated to establish the CP model.In control group, MPD was not ligated.The animals were killed in batches at 4th, 8th and 12th week after surgery.The pancreatic tissue was taken for elasticity modulus test and pathological examination, and the pigs were classified into control, mild, moderate and severe groups based on the severity of fibrosis.Cell density, fat infiltration and extracellular edema were observed and classified into mild and severe.The difference of elasticity modulus among different groups were compared by Variance analysis, the correlation between pancreatic fibrosis and elastic modulus was analyzed with Spearman correlation analysis, and ROC curve was used to evaluate its efficacy of diagnosing CP.Results Sixteen CP models were established successfully expected for 2 deaths (mild, n=7;moderate, n=2 and severe, n=7).All of the control group (n=3) showed normal pancreas.The elasticity modulus of control, mild and moderate to severe group were 0.4268±0.0566, 0.3203±0.0518 and 0.2235±0.0685, respectively.The difference between the groups was statistically significant (F=13.658,P<0.01), and the elastic modulus and pathological grade had a negative correlation (r=0.969, P<0.01).AUC of elasticity modulus for differentiating normal and mild CP was 1.000, the best critical value was 0.3807, and both the sensitivity and specificity of the diagnosis were 100%.AUC for differentiating mild and moderate to severe CP was 0.8730, the best critical value was 0.2646, and the sensitivity and specificity of the diagnosis were 85.7% and 77.8%, respectively.The pancreatic elasticity modulus of low parenchymal cell density group and high parenchymal cell density group were 0.1931±0.0373 and 0.3485±0.0655, respectively, which in the high cell density group was significantly higher than that in the low cell density group (t=-5.719, P<0.01).The elasticity modulus of negative infiltration or slight fatty infiltration group and severe fatty infiltration group were 0.3401±0.0697 and 0.1855±0.0344, respectively, which in the negative infiltration or slight infiltration group was significantly higher than that in severe infiltration group (t=5.102, P<0.01).The elasticity modulus of negative or mild cell edema group and moderate to severe cell edema group were 0.2760±0.0825 and 0.3024±0.1056, respectively;there was no statistically significant(t=-0.586, P >0.05).Conclusions The elasticity modulus can be used to detect the pathological changes of CP, and evaluate the CP pathologic grades.

BACKGROUND: Bone marrow-derived mononuclear cells (BM-MNCs) hold the potential of differentiating into osteoclasts. Polygonatum sibiricum polysaccharide (PSP) may inhibit the differentiation of BM-MNCs into osteoclasts and it is expected to become a new drug for the treatment of osteoporosis. OBJECTIVE: To investigate the effect of PSP on the differentiation of mouse BM-MNCs into osteoclasts induced by receptor activator of nuclear factor kappa-B ligand (RANKL) and bone resorption in vivo. METHODS: Mouse bone marrow-derived macrophages cultured in vitro, the effect of macrophage colony stimulating factor and PSP (5, 10, 20, 40, 80,160, 320, 640, 1280, 2560 mg/L) on the proliferation of mouse BM-MNCs was detected by cell counting kit-8 assay to determine the PSP concentration range; the mouse BMMs were cultured and induced in DMEM medium containing macrophage colony stimulating factor, RANKL and 5, 10, 20, 40, 80,160, 320, 640 mg/L PSP, respectively; those cultured without PSP served as control group. The morphological changes of cells were observed under an inverted microscope.; the number of osteoclasts was detected by tartrate-resistant acid phosphatase staining; the mRNA expression levels of osteoclast-related genes including tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 were evaluated by quantitative real-time PCR. A mouse model of calvarial osteolysis induced by lipopolysaccharide was established to receive PSP intervention, and then micro CT scanning, three-dimensional reconstruction and relevants software were used for quantitative analysis of bone volume/volume percentage, trabecular number, trabecular bone spacing and thickness. The number of osteoclasts was identified by tartrate-resistant acid phosphatase staining and quantitative analysis of bone resorption area was conducted. RESULTS AND CONCLUSION: Compared with the control group, the concentration of PSP below 640 mg/L showed no significant effect on the proliferation of BMMs (P > 0.05). Different concentrations of PSP (40-640 mg/L) significantly reduced the number of osteoclasts, osteoclast differentiation and maturation, and the mRNA expression levels of tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 TRAP, MMP-9, CtsK and NFATc1 (P < 0.05). Compared with lipopolysaccharide, PSP could effectively alleviate the lipopolysaccharide-induced calvarial osteolysis, and the bone volume/volume percentage, trabecular number, and trabecular bone spacing were significantly decreased (P < 0.05); additionally, the number of osteoclasts and the area of bone resorption were decreased significantly (P < 0.01). To conclude, PSP can inhibit the differentiation and maturation of mouse BMMs to osteoclasts and alleviate lipopolysaccharide-induced calvarial osteolysis.