BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) can be induced differentiating into osteoblasts and chondroblasts, DNA methylate depressant 5-azacytidine can induce BMMSCs expressing myogenic regulating factors: Myf5 and myopoietin, which involving in the differentiation of BMMSCs into myoblasts.OBJECTIVE: Muscular traumatic fluid containing the highest protein content was screened out and co-cultured with BMMSCs,in order to explore the inductive effect on the proliferation and myogeneis of BMMSCs.DESIGN : Standardized comparative study.SETTING .:At State Key Laboratory of Trauma, Bums and Combined Injury,Institute of Combined Injury, Medical College of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: Muscular traumatic model was established on 18 Balb/C pure rats for the extraction of muscular traumatic fluid, the inductive effect of the fluid on BMMSC was then compared with 5-azacytidine.METHODS:This study was carried out at State Key Laboratory of Trauma,Burns and Combined Injury, Institute of Combined Injury, College of Preventive Medicine, Third Military Medical University of Chinese PLA from April 2001 and September 2003. Bradford colorimetric was used to detect the protein content in the muscular traumatic fluid, and the fluid with the highest protein was used to co-culture with BMMSC, whose effect on the proliferation of BMMSC was measured with MTT methods at day 0, 3, 6, 9, 12, 15.RT-PCR technique was used to detect the expression of myopoietin at day 6,with its myogenic effect compared with that of 5-azacytidine.fluid on BMMSC.eration of BMMSC: the proloferative activity of BMMSC in traumatic fluid genic effect of traumatic fluid on BMMSC: myopoietin could not be found expressed in traumatic fluid group, but strongly expressed in 5-azacytidine group.CONCLUSION: Muscular traumatic fluid can promote the proliferation of BMMSC, but has no myogenic effect.

Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P<0.05). (2) The expression levels of PLAC4 and COL6A2 mRNA of the different pregnancy weeks of the healthy pregnancy group: the levels of PLAC4 mRNA in the peripheral blood of women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks of pregnancy were respectively 7.22 ± 1.05, 8.02±1.41,9.51±1.69,11.33±2.11 and 13.31±2.58, with their expression levels rising along with the increase of the pregnancy weeks; among them, the comparison of pregnancy 8 weeks and pregnancy 10 weeks, the variations had no statistical significance (P>0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.

【Objective】 To assess three severe acute respiratorysyndrome coronavirus 2 (SARS-CoV-2) enzyme linked immunosorbent assays (ELISA) and one pseudotype lentivirus-based neutralization test (ppNAT) in detecting the convalescent plasma antibody levles from COVID-19. 【Methods】 30 COVID-19 convalescent plasma samples were screened for antibodies against SARS-CoV-2 using three kinds of SARS-CoV-2 ELISA reagents and one ppNAT test in Shenzhen. The controls consisted of plasma samples from 32 healthy blood donors in February 2019. The diagnostic efficacy analysis of various SARS-CoV-2 ELISA reagents was performed using real-time fluorescent Polymerase Chain Reaction (RT-PCR). We also analyzed correlation between different immunological reagents and the age, gender, hospitalization, and severity of illness. 【Results】 The positive yielding rate of ppNAT and three kinds of IgG ELISA was higher than that of IgM ELISA. The positive yielding rates of three kinds of IgG ELISA were 100%(30/30), 93.33%(28/30), and 96.67%(29/30) respectively, while the yielding rates in control group were all 0. The positive yielding rate of three IgM ELISAs were 93.33%(28/30), 70%(21/30)and 46.67% (14/30). All the cases from negative control group were negative for IgG and IgM. Pearson correlation coefficient was calculated; there was a strong correlation between ELISA reagent 2 IgG and ELISA reagent 3 IgG (r=0.765, P<0.01). The correlation between ppNAT, ELISA reagent 3 IgG and the age of recovered patients was 0.422 and 0.385, respectively (P<0.05), while no significant correlation was found between the duration of hospitalization, severity of illness, gender and antibody signal/cutoff (S/CO) (P>0.05). 【Conclusion】 In the convalescent plasma with nucleic acid confirmed covid-19, the yielding rates of different IgM antibodies varied greatly. Antibody levels were influenced by age to some extent.

Humans ; Neoplasms, Second Primary/epidemiology* ; Neoplasms/epidemiology* ; Risk Factors ; Prognosis