Ambiguous results of the HLA-DPB1 genotyping results: Based on AllType NGS 11-loci sequencing reagent / 中国输血杂志

【Objective】 To statistically analyze the characteristics of ambiguous results of HLA-DPB1 genotyping given by AllType NGS 11-loci sequencing reagent via two next generation sequencing platforms, i. e. Ion Torrent S5 and Illumina Miseq. 【Methods】 A total of 434 samples from patients or donors were genotyped for HLA-DPB1 locus using AllType NGS 11-loci sequencing reagent from One Lambda company; 336 samples of them were sequenced via the Ion Torrent S5 platform and other 98 samples were sequenced via the Illumina Miseq platform. All 434 samples were genotyped for HLA-DPB1 gene simultaneously using PCR-SSO flow fluorescent bead method. The ambiguous genotypes of HLA-DPB1*13∶01∶01/107∶01 were distinguished by Sanger sequencing. The HLA-DPB1 genotype results by NGS method were assigned by TypeStream Visual professional software, and the ratio of ambiguous combination was calculated by direct count method. 【Results】 Ambiguous results were found in 357 out of 434 samples, accounting for 82.3% (357/434) when HLA-DPB1 allele was assigned to the third field using NGS method. Ambiguous results with 45 types were given in 275 out of 336 samples by the Ion Torrent S5 platform, accounting for 81.8% (275/336) and 82(with 27 types) out of 98 samples by the Illumina Miseq platform, accounting for 83.7% (82/98). All samples were re-genotyped for HLA-DPB1 gene by PCR-SSO, and none HLA-DPB1 allele had been missed by NGS. A total of 43 ambiguous alleles in HLA-DPB1*13∶01∶01/107∶01 involving 41 samples were distinguished by Sanger sequencing; HLA-DPB1*13∶01∶01 were detected in 25 (58.1%, 25/43) and HLA-DPB1*107∶01 in 18 (41.9%, 18/43). 【Conclusion】 There were still a high proportion of HLA-DPB1 ambiguous combinations using the AllType NGS 11-loci sequencing reagent. Sequencing exon 1 of HLA-DPB1 gene by Sanger sequencing can resolve part of the ambiguous results in HLA-DPB1*13∶01∶01/107∶01 alleles.

Analysis of loss of heterozygosity at HLA loci in a patient with leukemia / 中华医学遗传学杂志

OBJECTIVE@#To detect loss of heterozygosity (LOH) at human leukocyte antigen (HLA) loci in a Chinese patient with leukemia after haploidentical hematopoietic stem cell transplantation.@*METHODS@#HLA genotyping was carried out on peripheral blood, hair follicle and buccal swab samples derived from the patient after the transplantation as well as peripheral blood samples from his parents by using PCR-sequence specific oligonucleotide probe method and PCR-sequence based typing method. Short tandem repeat (STR) loci were detected by using a 23 site STR assay kit and a self-developed 6 STR loci assay for the HLA regions.@*RESULTS@#After the transplantation, the HLA genotype of the peripheral blood sample of the patient was identical to his father. The patient was HLA-A*02:01,24:02, C*03:03,03:04, B*13:01,15:01, DRB1*08:03,12:02, DQB1*03:01,06:01 for his hair follicle specimen. However, homozygosity of the HLA loci was found in his buccal swab sample. Only the HLA-A*24:02-C*03:03-B*15:01-DRB1*08:03-DQB1*06:01 haplotype from his father's was present, while the HLA-A*02:01-C*03:04-B*13:01-DRB1*12:02-DQB1*03:01 haplotype from his mother was lost. After the transplantation, the alleles of the 23 STR sites in the patient's peripheral blood sample were consistent to his father, with no allelic loss detected in his buccal swab sample. However, at least 4 STR loci in the HLA region were lost in his buccal swab sample.@*CONCLUSION@#LOH at the HLA loci has been detected in the buccal swab sample of a patient with leukemia who received haploidentical hematopoietic stem cell transplantation.

Assessment and risk factors analysis for quality of life in elderly patients with chronic kidney disease / 中华肾脏病杂志

Objective:To understand the comprehensive geriatric assessment (CGA) scores in chronic kidney disease (CKD) patients aged 65 years and older, and analyze the related influencing factors of quality of life.Methods:A total of 189 patients who were over 65 years old and diagnosed with CKD in the Department of Nephrology of Shanxi Provincial People's Hospital from October 2016 to October 2019 were included retrospectively. The patients were divided into dialysis group ( n=90 cases) and non-dialysis group ( n=99 cases) according to whether dialysis or not. The concise CGA scores included age, basic activities of daily living (BADL), instrumental activities of daily living (IADL), and modified cumulative illness rating score for geriatrics (MCIRS-G). Pearson correlation analysis was used to analyze the relationship between different scale scores and clinical indexes. Multiple linear regression analysis was used to further analyze independent related factors of the quality of life in elderly CKD patients. Results:Compared with the non-dialysis group, the BADL score and IADL score in the dialysis group were significantly reduced [(70.00±33.28) vs (93.38±14.32), t=6.166, P<0.001; (9.78±7.12) vs (15.95±5.74), t=6.520, P<0.001], while the MCIRS-G score was significantly increased [(31.13±4.00) vs (27.29±5.17), t=-5.741, P<0.001]. Linear regression analysis performed on the data of non-dialysis group patients showed that estimated glomerular filtration rate (eGFR), serum uric acid (SUA), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), blood potassium and chlorine were positively correlated with BADL and IADL scores (all P<0.05). B-type natriuretic peptide (BNP) was negatively correlated with BADL score ( P<0.01). BNP and age were negatively correlated with IADL score (both P<0.05). Fasting blood glucose (FBG) was positively correlated with MCIRS-G or MCIRS-G other than kidney (both P<0.05), and eGFR, SUA, total cholesterol, and HDL-C were negatively correlated with MCIRS-G or MCIRS-G other than kidney (all P<0.05). Multiple linear regression analysis showed that eGFR was an independent influencing factor for BADL ( P<0.01). Age and eGFR were independent influencing factors for IADL (both P<0.05). Conclusions:The decline of quality of life in elderly CKD patients is related with eGFR, SUA, age, BNP and HDL-C levels, and eGFR and age are independent influencing factors.

Mechanism of highly expressed primary cilia in tibial growth plate chondrocyte accelerating chondrocyte differentiation in young rats with chronic renal failure / 中华肾脏病杂志

Objective:To explore the mechanism of highly expressed primary cilia in tibial growth plate chondrocytes accelerating chondrocytes differentiation in young rats with chronic renal failure (CRF).Methods:Forty male 4-week-old SD rats weighing (98±3) g were randomly divided into control group (intragastric administration with distilled water, n=20) and CRF group (intragastric administration with adenine suspension 150 mg·kg -1·d -1, n=20). All the young rats were sacrificed after continuous gavage for 6 weeks. The length of the growth plate was measured with histological sections. Immunofluorescence (IF) was used to detect the expression rate of primary cilia and the level of β-catenin, the key protein of Wnt/β-catenin signaling pathway in tibial growth plate chondrocytes. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation, and the expression rate of primary cilia in chondrocytes, the levels of Indian hedgehog (IHH) and glycogen synthase kinase 3β (GSK3β) were detected by IF. Co-immunoprecipitation was used to detect the relationship between IHH and GSK3β. Results:Compared with the control group, the relative length of the growth plate was shorter in histological sections [(0.51±0.11) vs (1.00±0.08), t=16.11, P<0.001], the expression rate of primary cilia was higher [(26.3±5.5)% vs (7.6±1.9)%, t=14.37, P<0.001], and the level of β-catenin increased [(7.1±2.0) scores vs (3.6±1.0) scores, t=7.10, P<0.001] in CRF group. In vitro, the expression rate of primary cilia was higher in CRF group chondrocytes [(31.4±8.2)% vs (12.5±3.1)%, t=9.64, P<0.001] than that in control group. The level of IHH in CRF group increased than that in control group [(1 360±270) vs (310±84), t=16.61, P<0.001]. There was no significant difference in GSK3β level of chondrocytes between the two groups [(850±195) vs (780±140), t=1.30, P=0.200]. There was a direct interaction between IHH and GSK3β in CRF group chondrocytes. Conclusions:The expression levels of primary cilia and related protein IHH increase in tibial growth plate chondrocytes of CRF young rats. The IHH protein plays a direct interaction with GSK3β protein, Wnt/β-catenin signaling pathway antagonist, which leads to the activation of Wnt/β-catenin signaling pathway and final accelerated differentiation of chondrocytes. The rapid differentiation of chondrocytes causes the closing trend of growth plate.

Clinical and imaging features of 27 cases of childhood Sturge-Weber syndrome / 中华皮肤科杂志

Objective:To analyze clinical and imaging features of Sturge-Weber syndrome in children.Methods:Clinical data were collected from 27 children with Sturge-Weber syndrome in Xuzhou Children′s Hospital, Xuzhou Medical University from July 2013 to December 2019, and analyzed retrospectively.Results:Among the 27 children, 17 were males and 10 were females. Their age at the clinic visit ranged from 2 days to 10 years and 7 months, and averaged 2.54 years. All the 27 patients presented with facial port-wine stains of varied color from light red to purple red, which were all distributed across the facial midline, including 21 with predominantly unilateral port-wine stains and 6 with bilateral symmetrical port-wine stains. There were 17 patients with ocular choroidal vascular malformations, including 14 with congenital glaucoma, 5 with high intraocular pressure, and 1 with optic nerve atrophy accompanied by transient blindness. Neurological impairment occurred in 12 patients, and all manifested as epilepsy. All the 27 children underwent imaging examination, and abnormalities were found in 20. Among the 10 patients with abnormal computed tomography images, local calcification was observed in 8, and local thickening of the skull on the side affected by skin lesions in 8; 13 of 14 patients with abnormal magnetic resonance imaging scan results had signs of brain atrophy, 9 showed enhanced gyrus-like blood vessel formation by enhanced magnetic resonance imaging, and 5 showed decreased branches of the anterior and middle cerebral artery on the affected facial side by magnetic resonance angiography.Conclusions:Children with Sturge-Weber syndrome are clinically characterized by predominantly unilateral port wine stains on the face, some of whom are accompanied by epilepsy, glaucoma or mental retardation, and imaging examinations mainly show local calcification, brain atrophy, local thickening of the skull plate, enhanced gyrus-like blood vessel formation, etc. Early definite diagnosis and comprehensive systemic treatment are needed to reduce disability and mortality rates in patients with Sturge-Weber syndrome, and long-term follow-up should be considered.

Characteristics of microbiota of colonic lavage fluid from patients with diarrhea-predominant irritable bowel syndrome / 中华消化杂志

Objective To investigate the characteristics of the microbiota of colonic lavage fluid from patients with diarrhoea-predominant irritable bowel syndrome (D-IBS).Methods From August 2014 to March 2015,at the Department of Gastroenterology Shanxi Provincial People's Hospital,39 patients with D-IBS were enrolled and during the same period 39 patients with self-reported symptoms such as occasional abdominal discomfort or abdominal pain but were excluded from the diagnosis of irritable bowel syndrome (IBS) were selected as control group.The colonic mucosal tissues and colonic lavage fluid of two groups of patients were collected.DNA was extracted from tissues and colonic fluid.Ten dominant bacteria,including Bacteriodes prevotella,Bacteroidetes,Clostridium,Bifidobacterium,Dialister pneumosintes,Enteropathogenic Escherichia coli,Faecalibacterium prausnitzii,Firmicutes,Lactobacillus and Salmonella,were analyzed by real-time fluorescence quantitative polymerase chain reacticn (RT-PCR).T test and Spearman correlation analysis were performed for statistical analysis.Results The numbers of Bacteriodes prevotella in colonic lavage fluid and the descending colon mucosal tissues of patients in D-IBS group were 3.096 ± 1.462 and 4.278 ± 0.290,respectively,which were lower than those of control group (4.842 ± 1.032 and 5.341 ± 0.433,respectively),and the differences were statistically significant (t =6.093 and 12.657,both P ＜ 0.01).Correspondingly,the numbers of Bacteroidetes were 3.259 ± 0.607 and 3.698 ± 0.554,respectively,which were lower than those of control group (4.085 ± 0.504 and 4.339 ± 0.263,respectively),and the differences were statistically significant (t =6.538 and 6.528,both P ＜ 0.01).The numbers of Clostridium were 5.254 ± 0.522 and 4.471 ± 0.690,which were lower than those of control group (5.949 ± 0.505 and 4.950 ± 0.604,respectively),and the differences were statistically significant (t =5.976,P ＜ 0.0l;t =3.262,P =0.002).The number of Faecalibacterium prausnitzii from colonic lavage fluid of patients in D-IBS group was 2.238 ± 1.845,which was lower than that of control group (3.222 ± 1.450),and the difference was statistically significant (t =2.619,P =0.011).The Firmicutes (r =0.678,P =0.012),Lactobacillus (r =0.358,P =0.034),Bifidobacterium (r =0.418,P =0.042) and Faecalibacterium prausnitzii (r =0.568,P =0.022) from colonic mucosal tissues were all positively correlated with those from colonic lavage fluid.However,Salmonella (r =-0.638,P =0.012) and Enteropathogenic Escherichia coli (r =-0.343,P =0.025) were negatively correlated with those from mucosal tissues.Conclusions An imbalance of the microbiota in colonic lavage fluid is found in D-IBS patients.The numbers of Bacteriodes prevotella,Bacteroidetes,Clostridium and Faecalibacterium prausnitzii significantly decrease.

Vasorelaxant effect and mechanism of niclosamide ethanolamine on rat coronary artery / 中国比较医学杂志

Objective To investigate the vasorelaxant effect and possible mechanism of niclosamide ethanolamine (NEN)on isolated rat coronary artery(RCA). Methods Wire myograph was used to record myogenic tone of vessels. The vasorelaxant effect of NEN was studied in RCA precontracted with either KCl or U46619. Study of related inhibitors was performed to investigate possible involvement of potassium channels and the mitogen-activated protein kinase(MAPK) signaling pathway in vasorelaxation. The effect of NEN on Ca2+mobilization was determined by observing vasoconstrictor-induced contractions in tissue solution deprived of Ca2+followed by Ca2+restoration. Results NEN(0.5-3.0 μmol/L) relaxed RCA precontracted with KCl or U46619 in a concentration-dependent manner. MAPK inhibitors(PD98059 and SB239063)reduced the relaxant effect of NEN,while the potassium-channel blockers(tetraethylamine,4-aminopyridine, BaCl2,and glibenclamide)did not significantly affect relaxation. NEN specifically inhibited the contraction component dependent on extracellular Ca2+influx in vessels stimulated with KCl and U46619, with a negligible effect on the component dependent on intracellular Ca2+release. Conclusions NEN exhibits vasodilator properties in RCA. Inhibition of extracellular Ca2+influx and activation of the MAPK signaling pathway may be involved in NEN-induced RCA vasorelaxation.

Physiological response and subjective experience in evoked anger between feminine and masculine women / 中国心理卫生杂志

Objective:To measure the physiological response and subjective experience differences between feminine and masculine women inducing anger,exploring women's susceptibility to depression.Methods:Thirty feminine women and 26 masculine women were selected from 745 college women students who filled in the Chinese College Students'Sex Role Inventory-50 items (CSRI-50),according to median classification method.Physiological response,including heart rate,skin conduct,respiration rate were recorded using Biotrace software from 5-10 min before watching films to films end,state anger were measured before and after films by the State Anger Scale(SAS),one of the State-Trait Anger Expression Inventory-Ⅱ (STAXI-Ⅱ).Results:With inducing anger,the skin conducts were lower in feminine women than in masculine women[(6.1 ±4.6) μmho vs.(9.4 ±6.0)μmho,P ＜0.05],while the respiration rates were higher in feminine women than in masculine women[(19.4 ±2.8) bpm vs.(17.7 ± 3.3) bpm,P ＜ 0.05].The SAS sores were higher in feminine women than in masculine women[(31.6 ± 8.8) vs.(26.5 ± 9.2),P ＜ 0.05].Conclusion:It suggests that feminine women subjectively report more anger and physiological responses may be greater in watching anger inducing films,mainly in RSP rate,however masculine women's SC is higher,which indicates that future researches of anger and depression should consider sex role.

The correlation study between the changes of intestinal mucosa predominant bacteria and Toll-like receptor 2, Toll-like receptor 4 gene expressions in diarrhea-predominant irritable bowel syndrome patients / 中华内科杂志

Based on high throughput sequencing and PCR detection technology,this study has found out that intestinal microbial diversity was impaired and the quantities of two main bacteria flora (Bacteroidetes and Clostridium) were significantly reduced in patients with diarrhea-predominant irritable bowel syndrome (D-IBS).Meanwhile mucosal expression of toll-like receptor (TLR) 2 and TLR4 were significantly enhanced,which was inversely correlated with the reduction of Bacteroidetes and Clostridium.Thus,it suggests that D-IBS may be associated with TLR signal transduction triggered by the intestinal dysbacteriosis.

Effects of methylated CpG islands in promoter region on the expression of KIR3DL1 protein / 中华微生物学和免疫学杂志

Objective To investigate the effects of methylated CpG islands in the promoter region on the expression of killer cell immunoglobulin-like receptor 3DL1 (KIR3DL1).Methods Three voluntary unpaid blood donors carrying high expression allele KIR 3DL1*01502 and three donors carrying low expres-sion allele KIR3DL1*005 were recruited in this study .The nucleotide sequences and the methylated CpG islands in the promoter regions of KIR 3DL1*01502 allele and KIR3DL1*005 allele were analyzed .The NK cells expressing KIR3DL1*01502 and KIR3DL1*005 were respectively treated with 5-aza for the dem-ethylation of CpG islands within the promoters .The expression of KIR3DL1 protein on the surface of NK cells was measured with flow cytometer .Results Two differences at nucleotide sites -65 and -269 were detected within the promoter regions of KIR3DL1*01502 and KIR3DL1*005, resulting in two distinct CpG islands.The CpG islands within the promoter of KIR 3DL1*01502 allele were highly methylated .The ex-pression of KIR3DL1 protein on NK cells which carried KIR 3DL1*01502 allele was significantly increased after the demethylation of CpG islands .However , the treatment of demethylation had no significant effects on the expression KIR3DL1 protein on NK cells harboring KIR3DL1*005 allele.Conclusion The methylated CpG islands within the promoter of KIR 3DL1*01502 allele affected the antigen expression on the surface of NK cells.Different KIR3DL1 alleles might show different mechanisms in regulating antigen expression .

A clinical study of chromosome translocations in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue in Chinese patients / 中华内科杂志

Objective To investigate the genetic aberrations in extranodal marginal zone lymphoma of mueosa-associated lymphoid tissue (MALT) lymphomas from different sites of the body in Chinese patients. Methods Two hundred and seventeen paraffin-embedded MALT lymphoma specimens from 11 major sites were studied with interphase fluorescence in situ hybridization (FISH) to detect t(11; 18) (q21;q21)/API2-MALT1, t(1; 14) (p22; q32)/IGH-BCL10, (14; 18) (q32; q21)/IGH-MALT1 and BCL6 gene involved chromosome translocations. Results These translocations were mutually exclusive and detected in 21% (46/217) of the cases, including t(11;18) (q21;q21) API2-MALT1 13% (29/217), t (1;14)(p22 ;q32) IGH-BCLIO in 1% (3/217), t(14;18) (q32;q21) IGH-MALT1 1% (2/217), BCL6 involved translocation in 2% (4/217) and IGH-unknown translocation partner in 4% (8/217). t(11; 18) (q21;q21)API2-MALT1 was found with the highest frequency in MALT lymphoma from lungs (47% , 8/17) and small intestine (29%, 4/14), followed by salivary gland (17%, 1/6), stomach (14%, 12/84) and ocular adnexae (6% , 4/68). t(1 ;14) (p22;q32) was only detected in lungs (12%, 2/17) and stomach (1%, 1/84). t(14;18) (q32;q21) was mainly detected in lungs (6%, 1/17) and ocular adnexae (2%, 1/68). BCL6 gene involved translocation was detected in salivary gland (17% , 1/6) and stomach (4%, 3/84). Conclusions It is demonstrated that the four translocatidns occur with markedly variable frequencies in MALT lymphoma of different sites in Chinese patients. The distributions of these chromosome translocations in Chinese patients are slightly different from those reported in western patients.