The present study examined the role of Wnt/β-catenin signaling pathway in the degeneration of nucleus pulposus cells and the protective effect of DKK1 on nucleus pulposus cells. The model of nucleus pulposus cell degeneration was induced by intra-disc injection of TNF-α, and the expression of β-catenin protein was detected by Western blotting. The cultured rabbit nucleus pulposus cells were divided into 4 groups. In group A, the cells were cultured with normal medium and served as control group. In group B, the cells were cultured with TNF-α and acted as degeneration group. In group C, the cells were cultured with TNF-α and transfected with Adv-eGFP and was used as fluorescence control group. In group D, the cells were cultured with TNF-α and transfected with Adv-hDKK1-eGFP, serving as intervention group. The expression of type II collagen, proteoglycan, β-catenin, and MMP-13 in each group was detected by immunocytochemistry and RT-PCR. The result showed that TNF-α increased the expression of β-catenin and MMP-13, and significantly inhibited the synthesis of type II collagen and proteoglycan, which resulted in the degeneration of nucleus pulposus cells. This effect could be obviously reversed by DKK1. We are led to concluded that TNF-α could activate the Wnt/β-catenin signaling pathway, and increase the expression of MMP-13, thereby resulting in disc degeneration. Specifically blocking Wnt/β-catenin signaling pathway by DKK-1 could protect the normal metabolism of intervertebral disc tissue. The Wnt pathway plays an important role in the progression of the intervertebral disc degeneration.

OBJECTIVEA multiplex PCR system for screening rare blood group antigens K and Yt(b) was constructed to study the distribution of the two blood groups in a Uygur population in Xinjiang, China.

METHODSSequence-specific primers (SSP) were designed based on single nucleotide polymorphism sites of KEL and ACHE alleles encoding the two blood group antigens. The system was designed for simultaneously detecting the two antigens by optimizing the PCR reaction. Three hundred and sixty-two randomly selected healthy individuals were screened. Products of PCR were further analyzed for heterozygosity.

RESULTSThe system was set up successfully. No KK sample was identified and 9 K+ k+ , 41 Yt (a+ b+ ), 4 Yt (a- b+ ) were found among the 362 samples.

CONCLUSIONThe established PCR-SSP based multiple PCR system is efficient to screen the rare blood group antigens K and Yt(b). The information of rare blood donors obtained from the screening can be used to improve the capability of compatible transfusion.

Antigens, Bacterial ; genetics ; Antigens, Surface ; genetics ; Blood Donors ; Blood Group Antigens ; genetics ; Blood Transfusion ; methods ; China ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide

Objective: To construct ¹³¹I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP; SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR; GP) and double targeting peptide SR/GP, and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model. Methods: PAMAM(G5.0), PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP were radiolabeled with ¹³¹I by chloramine T method. The radiolabeled yield and radiochemical purity were determined by thin layer chromatography. MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (%ID/g) in tumor and organs was measured at 24 h post-injection. Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4, 8 and 24 h post-injection were calculated and compared among different groups. One-way analysis of variance, repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups. The relationship between %ID/g and T/NT was analyzed with Pearson correlation. Results: The radiolabeled yield was more than 75% and radiochemistry purity was more than 90%. The difference of %ID/g at 24 h post-injection was significant (F=14.400, P<0.001) in tumors of all groups. The radioactive uptake in tumor of ¹³¹I-PAMAM(G5.0)-SR group was the highest at 24 h post-injection((1.80±0.18) %ID/g). There were significant differences of T/NT ratios among different groups(F=4.776, P<0.05)and between different time points(F=8.630, P<0.05). Compared with negative control group (Na¹³¹I), the T/NT ratios significantly increased in ¹³¹I-PAMAM(G5.0)-SR group at 4, 8 and 24 h post-injection (t=4.169, 7.123 and 4.032, all P<0.05) and in ¹³¹I-PAMAM(G5.0)-GP group at 4 h post-injection (t=5.893, P<0.05). The T/NT ratio in ¹³¹I-PAMAM(G5.0)-SR group was higher than that in ¹³¹I-PAMAM(G5.0)-GP group at 24 h post-injection (t=2.871, P<0.05). Conclusions PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models. ¹³¹I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis, treatment and prognosis evaluation.

Objective:To investigate the efficacy of Sishen pill compound combined with mesalazine in the treatment of mild to moderate ulcerative colitis and its effect on the circadian rhythm of symptoms. Methods:A total of 136 patients with mild to moderate ulcerative colitis who received treatment in Hospital of Chengdu University of Traditional Chinese Medicine from January 2018 to December 2020 were included in this prospective randomized controlled trial. These patients were divided into a treatment group ( n = 68) and a control group ( n = 68). The treatment group was treated with Sishen pill compound combined with mesalazine. The control group was treated with mesalazine alone. All patients were treated for 12 weeks. Clinical efficacy, as well as morning abdominal pain grade, morning diarrhea score, fecal trait score, Mayo score, hemoglobin, and hypersensitive C-reactive protein pre- and post-treatment, were compared between the two groups. Results:Total response rate in the treatment group was significantly higher than that in the control group [91.18% (62/68) vs. 72.06% (49/68), χ2 = 8.28, P < 0.05]. After treatment, morning diarrhea score, morning abdominal pain score, fecal trait score, Mayo score, hemoglobin, and hypersensitive C-reactive protein in the treatment group were (0.47 ± 0.56) points, (0.53 ± 0.56) points, (3.01 ± 0.72) points, (7.13 ± 1.38) points, (108.04 ± 12.21) g/L, (4.00 ± 2.19) mg/L, respectively, and they were (0.84 ± 0.56) points, (1.12 ± 0.56) points, (4.40 ± 0.76) points, (3.25 ± 1.44) points, (102.15 ± 12.61) g/L, and (6.07 ± 3.66) mg/L respectively in the control group. There were significant differences in these indexes between the treatment and control groups ( t = 3.59, 5.95, 10.06, 9.62, 2.78, 3.99, all P < 0.05). Conclusion:Sishen pill compound combined with mesalazine can effectively reduce clinical symptoms of active ulcerative colitis, increase hemoglobin level, decrease C-reactive protein level, improve the efficiency of treatment, reduce symptoms and the number of diarrhea rhythms, and improve stool symptoms of mild to moderate ulcerative colitis patients.

The present study examined the role of Wnt/β-catenin signaling pathway in the degeneration of nucleus pulposus cells and the protective effect of DKK1 on nucleus pulposus cells.The model of nucleus pulposus cell degeneration was induced by intra-disc injection of TNF-α,and the expression of β-catenin protein was detected by Western blotting.The cultured rabbit nucleus pulposus cells were divided into 4 groups.In group A,the cells were cultured with normal medium and served as control group.In group B,the cells were cultured with TNF-α and acted as degeneration group.In group C,the cells were cultured with TNF-α and transfected with Adv-eGFP and was used as fluorescence control group.In group D,the cells were cultured with TNF-α and transfected with Adv-hDKK1-eGFP,serving as intervention group.The expression of type Ⅱ collagen,proteoglycan,β-catenin,and MMP-13 in each group was detected by immunocytochemistry and RT-PCR.The result showed that TNF-α increased the expression of β-catenin and MMP-13,and significantly inhibited the synthesis of type Ⅱ collagen and proteoglycan,which resulted in the degeneration of nucleus pulposus cells.This effect could be obviously reversed by DKK1.We are led to concluded that TNF-α could activate the Wnt/β-catenin signaling pathway,and increase the expression of MMP-13,thereby resulting in disc degeneration.Specifically blocking Wnt/β-catenin signaling pathway by DKK-1 could protect the normal metabolism of intervertebral disc tissue.The Wnt pathway plays an important role in the progression of the intervertebral disc degeneration.

Evidence-based medicine (EBM) is a kind of clinic practice where clinicians use the best and the latest available evidence to diagnose and treat patients, and both evidence providers and users need to identify and control different kinds of biases in medical research.Directed acyclic graphsis is a tool to explore the causal relationship.The possible biases in the study can be revealed in a simple graphical language.The use of directed acyclic graphs could avoid the occurrence of bias and improve the quality of medical research and better guide clinical practice.

OBJECTIVETo screen rare blood groups Fy(a-), s-, k-, Di(b-) and Js(b-) in an ethnic Zhuang population.

METHODSSequence-specific primers were designed based on single nucleotide polymorphism (SNP) sites of blood group antigens Fy(b) and s. A specific multiplex PCR system I was established. Multiplex PCR system II was applied to detect alleles antigens Di(b), k, Js(b)1910 and Js(b) 2019 at the same time. The two systems was were used to screen for rare blood group antigens in 4490 randomly selected healthy donors of Guangxi Zhuang ethnic origin.

RESULTSWe successfully made the multiplex PCR system I. We detected the rare blood group antigens using the two PCR system. There are five Fy(a-), three s(-), two Di(b-) in 4490 Guangxi zhuang random samples. The multiplex PCR system I has achieved good accuracy and stability. With multiplex PCR systems I and II, 4490 samples were screened. Five Fy(a-), three s(-) and two Di(b-) samples were discovered.

CONCLUSIONMultiplex PCR is an effective methods, which can be used for high throughput screening of rare blood groups. The rare blood types of Guangxi Zhuang ethnic origin obtained through the screening can provide valuable information for compatible blood transfusion. Through screening we obtained precious rare blood type materials which can be used to improve the capability of compatible infusion and reduce the transfusion reactions.

Blood Group Antigens ; genetics ; Duffy Blood-Group System ; genetics ; Genotype ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Receptors, Cell Surface ; genetics