Evaluation on the application of light scattering method to detect hemoglobin in lipoblood samples / 中华检验医学杂志

Objective:To evaluate the ability of light dispersive colorimetry to detect hemoglobin (Hb) in lipid blood samples and its feasibility as an alternative to plasmapheresis commonly used in the laboratory.Methods:Routine blood samples of 276 inpatients in Fujian Provincial Hospital from July 2020 to July 2021 were collected. Routine blood samples of 276 inpatients were collected. There were 169 males and 107 females, aged from 16 to 97 years. 183 non-lipid blood samples and 93 lipid blood samples were collected.(1) One case each of low, medium and high Hb value in non-lipid blood and lipid blood samples were collected, and the precision and the linearity of light scattering method was detected.(2)Non-lipid blood samples divided into Hb low-value group, median-value group and high-value group, which were measured by light scattering method and colorimetric method to compare Hb values. (3)Non-lipid blood samples were divided into Hb low-value group, median-value group and high-value group. Plasma exchange was carried out with different concentrations of fat emulsion. The bias and linearity of Hbc2 and Hbc1, Hbo2 and Hbc1 were analyzed by MedCalc19.1 software. The Hbc2 and Hbc1 bias ( CV%) and Hbo2 and Hbc1 bias ( CV%) were calculated. T test was used to analyze the influence of different concentrations of triglyceride on Hb bias.(4)Blood samples were divided into Hb low-value group, median-group and high-value group. The Hb of light scattering method was compared with the colorimetric method after plasma exchange. Results:(1)The intrabatch precision of light scattering method for non-lipid blood and lipid blood specimens was within the allowable range ( CV<1.5%), and the good linearity ( R2=1.000).(2)The bias of Hb measured by light scattering method and colorimetric method in the three groups was below 3.5%(-0.58±2.34,0.16±1.52,1.15±1.56), within the allowable total error range. The two methods have the equivalence and good linear relationship ( r=0.999).(3)The concentrations of Hbo2 in the low (except 4.1 mmol/L), medium and high Hbo2 groups were equivalent to those in the non-lipid blood colorimetry (Hbc1), and the two methods were well correlated. The results of light scattering method have nothing to do with the concentration of lipid blood.(4)There was no significant difference of the Hb between the light scattering method and plasma exchange method in three groups ( P>0.05), Both of them have equivalence and good correlation ( R2=0.968,0.948,0.870). Conclusion:Light scattering method can effectively reduce the effect of lipid blood on hemoglobin determination, and can replace the traditional plasma exchange method, which has high clinical application value.

Role and mechanism of PRMT4 in genesis and progression of gastric cancer / 上海交通大学学报（医学版）

Objective • To investigate the effect of protein arginine methyltransferase 4(PRMT4) on the development and progression of gastric carcinoma and the underlying molecular mechanisms. Methods • The expression levels of PRMT4 in 32 specimens of gastric cancer and adjacent tissues were measured by immunohistochemistry technology. Public databases were used to analyze the expression of PRMT4 in gastric cancer and its effect on the survival of patients with gastric cancer. PRMT4 expressions of gastric cancer MGC-803 and MKN-45 cells were down-regulated by short hairpin RNA-mediated knockdown, and PRMT4 expression of gastric cancer SGC-7901 cells was up-regulated by using plasmid-mediated overexpression. Cell biology methods, such as MTT assay, colony formation assay, flow cytometry, and Transwell assay were used to study the effect of PRMT4 on proliferation, apoptosis, migration and invasion of gastric cancer cells. Luciferase reporter assay was used to detect transcription activity of endogenous transcription factor TCF (T cell factor) after PRMT4 silencing. The mRNA level changes of Wnt/β-catenin pathway downstream target genes after PRMT4 silencing were detected by real-time PCR. The protein level changes of Wnt/β-catenin pathway-related proteins after PRMT4 silencing and overexpression were detected by Western blotting. Results • PRMT4 was highly expressed in gastric cancer tissues. Patients with higher expression of PRMT4 had shorter survival than those with lower expression of PRMT4. The proliferation, colony formation, migration and invasion ability of gastric cancer cells were inhibited; the cell apoptosis was induced by silencing of PRMT4. The Wnt/β-catenin signaling pathway was also inhibited by PRMT4 knockdown. The proliferation, colony formation, migration and invasion ability of gastric cancer cells were induced; the cell apoptosis was inhibited by overexpressed PRMT4. The Wnt/β-catenin signaling pathway was also activated by PRMT4 overexpression. Conclusion • PRMT4 is highly expressed in gastric cancer. It can promote the growth, migration and invasion of gastric cancer cells through Wnt/β-catenin signaling pathway.

The correlation between atypical respiratory pathogens infection and serum total IgE levels / 国际检验医学杂志

Objective To investigate the correlation between atypical respiratory pathogens infection and serum total IgE levels . Methods Serum IgM level was detected in 1 913 blood samples of children with atypical respiratory infection by using indirect im‐munofluorescence assay ,including mycoplasma pneumonia(MP) ,legionella pneumophila (LP) ,rickettsia Q(QFR) ,chlamydia pneu‐monia(CPn) ,adenovirus (Adv) ,respiratory syncytial virus (RSV) ,influenza A virus (IAv) ,influenza B virus (IBv) and parainflu‐enza virus (PIV)1/2/3 .The serum total IgE level was detected by immune scatter turbidimetry .Software SPSS 17 .0 was used in data statistical analysis .Results A total of 991 out of 1 913 samples of respiratory inflected children exhibited positive(positive group) ,while 922 exhibited negative(negative group) in indirect immunofluorescence assay .650 out of the 991 positive samples (65 .59% ) contained MP infection and the combination of MP infection and other virus infections .The serum total IgE level in posi‐tive group was significantly higher than that of the negative group ,and the serum total IgE level in samples with MP infection was higher than that in samples with IBv infection ,Adv infection ,and RSV infection .In the samples in which serum total IgE level was higher than the clinical reference range (100 kU/mL) ,the infection rate of MP infection alone was 31 .29% ,which was evidently higher than that in samples of low IgE level(< 100 kU/mL ,21 .30% ) .On the other hand ,the infection rate of RSV alone was 1 .88% and the infection rate of Adv alone was 3 .13% ,which were both evidently lower than those in samples with normal serum total IgE level(both 6 .53% ) .Conclusion MP is the most common pathogen in children with atypical respiratory pathogen infec‐tion ,and can lead to higher serum total IgE levels .

Mutation analysis of FAH gene in patients with tyrosinemia type 1 / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical features and mutations of the FAH gene.</p><p><b>METHOD</b>Clinical records of two cases were collected, and diagnosis was made according to the diagnostic criteria of the International Organization for Rare Disorders (NORD). Genomic DNA was extracted from peripheral blood leukocytes with QIAamp DNA Mini Kit. The DNA extracts were subjected to direct sequencing for 14 exons together with adjacent fragments of FAH gene using ABI Prism 3730 Genetic Analyzer (Applied Biosystems, Foster City, CA) after PCR based on genomic DNA. The mutation source was verified by analyzing parents' exons corresponding to patients' mutation exons. The homology between human FAH enzyme and that of other species was surveyed using software Clustal X(European Bioinformatics Institute, Hinxton, Saffron Walde, UK). Polyphen (Polymorphism Phenotyping), available online, were used to predict possible impact of an amino acid substitution on structure and function of FAH enzyme. Polyphen calculates position-specific independent counts (PISC) scores for two amino acid variants in polymorphic position. A PISC scores that differ by > 2 were regarded as indicating the probability of damaging variants.</p><p><b>RESULT</b>Patient 1 was a 5 months and 21 days-old boy who suffered from persistent diarrhea, hepatomegaly, ascites; Alpha-fetoprotein > 1210 µg/L, levels of tyrosine in blood and succinylacetone in urine were 110.8 µmol/L and 83.7 µmol/L. His sister suffered from tyrosinemia type 1. Direct sequencing showed a G to A transition in CDS position 455 and 1027. He was compound heterozygous for the mutation c.455G > A/c.1027G > A, which predicts a change from tryptophan to a stop codon (TGG > TAG) at position 152 (W152X) and a change from glycine to arginine (GGG > AGG) at position 343 respectively. Patient 2 was a 6 year and 1 month-old girl with late-onset rickets who had signs of hepatosplenomegaly, rachitic rosary, windswept knees. Hypophosphatemia and alkaline phosphatase 1620 IU/L were detected. Alpha-fetoprotein 412.8 µg/L, levels of tyrosine in blood and succinylacetone in urine were 835.8 µmol/L and 27.48 µmol/L. Rickets did not improve after administration of calcium and vitamine D3. She is homozygous for the mutation c.1027G > A/c.1027G > A, which predicts G343R. The parents were mutation carriers. Analysis by Clustal X on the alignment of amino acids residual reservation among different species showed that the locative amino acid was highly conserved. Polyphen software predicted G343R was probably damaging (PISC score 3.235).</p><p><b>CONCLUSION</b>Children with tyrosinemia type 1 can have manifestations of persistent diarrhea or late-onset rickets. Physical examination can reveal hepatosplenomegaly, laboratory tests indicate markedly elevated serum concentration of alpha-fetoprotein and alkaline phosphatase in plasma and succinylacetone in urine, other members in family may have tyrosinemias or parents are consanguineous. Mutations c.455G > A and c.1027G > A can be detected in FAH gene of Chinese children.</p>

Research and synthesis of organosilicon nonthrombogenic materials containing sulfobetaine group.

A novel organosilicon sulfobetaine was synthesized through the reaction of organosilicon containing tertiary amino with 1,3-propanesulfone. Then this organosilicon sulfobetaine was coated onto polyurethane and organosilicon surface to improve their blood compatibility. The existence of sulfobetaine structure on the surface of materials was revealed by ATR-FTIR and XPS. The thermo-capability of synthesized silicone rubber, containing sulfobetaine was revealed by TGA. The blood compatibilities of organosilicon sulfobetaine and other materials such as silicone and PU as reference coated with them were evaluated by platelet-rich plasma adhesion experiment. The novel segmented silicone rubber containing sulfobetaine structure showed perfect blood compatibility.

Construction of the Expression Vector of Viruslike Particles Containing FMDV IRES RNA / 中国生物工程杂志

The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

Microcalorimetric study on host-guest complexation of naphtho-15-crown-5 with four ions of alkaline earth metal / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Thermodynamic parameters of complexation of naphto-15-crown-5 with four alkaline earth ions in aqueous media was determined using titration microcalorimetry at 298.15 K. The stability of the complexes, thermal effect and entropy effect of the complexation is discussed on the basis of the guest ions structure and the solvent effect. The stability constants tendency to vary with ion radius was interpreted. Complex of naphtha-15-crown-5 with calcium ion is very stable due to the synergism of static electric interaction and size selectivity between the host and the guest.