Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development / 中华儿科杂志

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.

Genetic susceptibility of ABCC4, FCGR2A and BLK polymorphisms with Kawasaki disease in children from southern China / 中华检验医学杂志

Objective To investigate whether single nucleotide polymorphisms(SNPs)in the ABCC4(rs7320375,rs7329490,rs7986087),FCGR2A(rs1801274)and BLK(rs2254546)region could be susceptibility locus for Kawasaki disease(KD)in children from southern China.Methods This study was performed as a case-control study.The samples,92 individuals with KD and 194 healthy controls from southern China,were collected at Guangzhou Women and Childrens′Medical Center from October,2013 to November,2015,and the SNPs were genotyped by using the Sequenom MassArray system.The genotype distribution and allele frequency of the SNPs were analyzed using chi-square test and Fisher′s exact test.Results The genotype distribution of FCGR2A(rs1801274)in patients with KD were as follows: GG 4.3％(4/92),AG 33.7％(31/92),AA 62％(57/92),correspondingly in healthy controls were GG 48.5％(94/194),AG 41.2％(80/194),AA 10.3％(20/194)respectively,and a significant difference was found between KD patients and controls(x2=98.17,P=0.000).A allele frequency of FCGR2A(rs1801274)in KD patients(78.8％,145/184)was higher than that in controls(30.9％,x2=0.120,P=0.000).The genotype distribution of BLK(rs2254546)in patients with KD were as follows: GG 67.4％(62/92),AG 28.3％(26/92),AA 4.3％(4/92),correspondingly in healthy controls were GG 52.1％(101/194),AG 43.8％(85/194),AA 4.1％(8/194)respectively,significant differences were found between KD patients and controls(x2=6.47,P=0.039).G allele frequency of BLK(rs2254546)in KD patients(81.5％,150/184)was higher than that in controls(74.0％,287/388,x2=1.553,P=0.047).Conclusions For the children in southern China,FCGR2A SNPs(rs1801274)may be associated with the susceptibility to KD,and the A allele may increase the risk of KD.BLK SNPs(rs2254546)is also found to be associated with the susceptibility to KD,and the G allele may increase the risk of KD.

Preliminary study on value of lymphocyte function detection in diagnosis and therapy of Henoch-Schonlein purpura / 国际检验医学杂志

Objective To investigate the changes of peripheral blood lymphocyte function in the children patients with acute phase of Henoch-Schonlein purpura(HSP),and to explore its significance in the diagnosis and therapy of HSP.Methods The peripheral blood lymphocytes subsets CD3+,CD4+,CD8+,CD4+/CD8+,CD19+ and NK cells(CD16++ CD56+) were detected in 51 children patients with acute phase of HSP and 40 children undergoing physical examination by flow cytometry(FCM Results Compared with the control group,the counts of peripheral blood CD4+,CD4+/ CD8+and NK cells CD16++ CD56+) in the acute stage of the HSP group were significantly decreased(P<0.01),while the proportions of CD8+ and CD19+cells were significantly increased(P<0.05,P<0.01).Conclusion The immunity dysfunction exists in the children patients with acute phase of HSP.The peripheral blood lymphocyte function detection has an important significance for the early diagnosis and treatment of HSP.

In vitro inhibitory effect of artesunate on the vascular endothelial growth factor secreted by mononuclear cells in hand-foot-and-mouth disease children complicated with encephalitis / 中华传染病杂志

Objective To investigate the dynamics of white blood cell count (WBC),prealbumin(PA),high sensitive C-reactive protein (hs-CRP),and vascular endothelial growth factor (VEGF) in hand-foot-and-mouth disease (HFMD) complicated with or without encephalitis and in vitro inhibitory effects of artesunate on VEGF secretion of mononuclear cells from HFMD patients.Methods Peripheral blood mononuclear cells from healthy control group,HFMD group and HFMD combined with encephalitis group were isolated by Ficoll density gradient centrifugation and treated with different concentrations of artesunate (25,50,100 mg/L).The expression of VEGF in the supernatant was examined by ELISA double antibody sandwich method.The levels of WBC,PA and hs-CRP in the three groups were also detected.Multiple samples were compared by one-way analysis of variance.Multiple comparison was performed by Dunnett T3 test.Correlation of two variables was analyzed by Spearman correlation test.Results There were significant differences in the levels of WBC,hs-CRP,PA and VEGF between the HFMD combined with or without encephalitis group and health control group (F=172.69,366.02,166.32 and 5 941.89,respectively,all P＜0.01).There were significant differences in the levels of VEGF secreted by mononuclear cells treated with three different concentrations of artesunate between the HFMD combined with or without encephalitis groups and health control group (F =194.265 and 4 750.69,respectively,both P＜0.01).Correlation analysis showed that the VEGF level secretion by mononuclear cells from HFMD patients with or without encephalitis were both negatively correlated with different concentrations of artesunate (r=-0.903 and-0.969,both P＜0.01).Conclusions Compared with HFMD without encephalitis,the secretion of VEGF by mononuclear cells,WBC and hs-CRP levels in HFMD complicated with encephalitis group all increase and PA decrease significantly.Artesunate can inhibit the secretion of VEGF by mononuclear cells of patients with hand-foot-and-mouth disease in a dosedependent manner in vitro.

Inhibition of piperlongumine on platelet aggregation and blood coagulation for children in vitro study / 国际检验医学杂志

Objective To observe the inhibition of piperlongumine in vitro on platelet aggregation and blood coagulation tests for children,to provide the experimental basis for clinical medication.Methods Venous blood samples from 30 children were randomly devided into 5 groups,and was centrifuge to separate platelet-rich plasma (PRP).After storing in 37 ℃ thermostat water bath for 5 minutes,the PRP which have been added DMSO as blank group,and added Aspirin (10 μmol/L)as control group,and added PL (20 μmol/L),PL(100 μmol/L),PL(200 μmol/L) as different concentrations of PL groups respectively,were induced by the addition of adenosine diphosphate (10 μmol/L),collagen(2.5 μg/mL) and the arachidonic acid(500 μg/mL).Then the platelet aggregation rate of the PRP from 5 groups could be measured by turbidimetry.Blood plasma isolated from venous blood was divided into 5 groups.In the PL groups,blood plasma were mixed up with PRP concentrations of which were 5,10,20 μmol/L.In the bland group,blood plasma were mixed up with DMSO (1％).In the control group,blood plasma were mixed up with heparin sodium(35 U/mL).After storing in 37 ℃ thermostat water bath for 5 minutes,fibrinogen(FIB),prothrombin time(PT),thrombin time(TT) and activated partial thromboplastin time of different groups were detected.Results Compared to the control group,the groups which were add PL with different concentrations (20,100,200 μmol/L) showed significant inhibition on platelet aggregation induced by AA and collagen(P<0.05).PL with concentrations of 100 μmol/L and 200 μmol/L showed significant inhibition on platelet aggregation induced by ADP(P<0.05).The PT,APTT,TT of blood plasma from children had been significantly prolonged by the intervention of PL 10 μmol/L and PL 20 μmol/L(P<0.05),however,no significant change of FIB was observed.Conclusion There are inhibitory effects of PL on platelet aggregation of blood plasma from children and anticoagulant activity in this study.

Study on transdifferentiation-acquiring tissue stem cell potency during renal tubular epithelial cells inflammatory damage / 国际检验医学杂志

Objective To study the potency of transdifferentiated renal tubular epithelial cells for acquiring the tissue stem cells during renal fibrosis.Methods The in vitro cellular model of renal tubular epithelial cells(NRK-52E)transdifferentiation under the inflammatory environment of the local renin-angiotensin (AngⅡ)system was established.The expression and change situation of the embryonic kidney developmental gene Pax2 and the tissue stem cell surface marker CD133 were observed.Results Local high concentration of AngⅡcould stimulate the NRK-52E cells to express Pax2 and CD133 molecule,its effect demonstrated the dose-and time-dependent relation.Conclusion The inflammatory damage leads to the transdifferentiated renal tubular epithelial cells po-tency to acquire the tissue stem cell.

Presence of antibodies to cyclic citrullinated peptides in juvenile-onset systemic lupus erythematosus / 中华风湿病学杂志

Objective To determine the prevalence of antibodies to cyclic citrullinated peptides (antiCCP) in patients with juvenile-onset systemic lupus erythematosus (JSLE) and its potential clinical significance. Methods Anti-CCP was measured in sera from patients with JSLE (n=47), juvenile idiopathic arthritis (JIA, n=54) and the sera from age-matched healthy children (n=40) using the third generation of anti-CCP ELISA commercial kit. The association of anti-CCP with other laboratory parameters and clinical features, especially arthritic symptoms in JSLE was also analyzed. T-test, Mann-Whitney U test, Chi-square and Fisher's exact test were used for statistical analysis. Results Out of the 47 JSLE patients, 6 (13%) were anti-CCP positive, which was significantly higher than that of the healthy controls( 13% vs 0, P＜0.05 ), but not different from that of the JIA group (26%, P=0.098). RF was more prevalent in JSLE patients with anti-CCP than patients without (83% vs 15%, P＜0.01 ), but there was no difference in other laboratory parameters and the clinical features ineluding the occurrence of arthritis (67% vs 51%, P＞0.05). As one of the initial symptoms, arthritis was observed in 25 of 47 JSLE patients and no one had developed deforming arthropathy.There was no statistical difference in anti-CCP positivity between JSLE patients with and without articular involvement ( 16% vs 9%, P＞0.05 ). Anti-CCP was not detected in any of the 3 patients with JSLE who had experienced joint pain and limited activity during 3 years follow-up. Conclusion Anti-CCP could be detected in patients with JSLE. It is noteworthy when differentiate from juvenile idiopathic arthritis, but the presence of anti-CCP does not relate with the occurrence of arthritis at presentation and persistence of arthritis in JSLE.

Role of angiotensin II and JAK2 signal pathway in transdifferentation of renal tubular cells in mice after acute ischemic followed by reperfusion / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of angiotensin (Ang)II and its Janns-activated kinase-2 (JAK2) signal pathway in transdifferentiation of renal tubular cells under the challenge of acute ischemic reperfusion injury.</p><p><b>METHODS</b>Models of acute ischemic reperfusion injury were established and the level of local AngII, a key element of renin-angiotensin system (RAS), in kidney was measured using radioimmunity technique. The expression of alpha-smooth muscle actin (alpha-SMA), a phenotype of mesenchymal cells, was detected by RT-PCR and immunohistochemistry methods. Renal tubule cells (NRK-52E) were cultured with various concentration of AngII, followed by blocking of PD123319, AngII receptor 2 antagonist, and AG490, an inhibitor of JAK2 signal pathway.</p><p><b>RESULTS</b>AngII of kidney tissue increased immediately after acute ischemic-reperfusion injury, in time dependent fashion. Expression of alpha-SMA in renal tubule cells was found at 48 hours after ischemic-reperfusion injury and in NRK-52E cells treated by high concentration of AngII and was dose and time dependent. The peak of alpha-SMA expression was seen after 30 minute treatment at the dose of 10(-9) mol/L, which was interrupted by both of PD123319 and AG490.</p><p><b>CONCLUSIONS</b>Transdifferentiation of renal tubular epithelial cells occurs under acute ischemic-reperfusion injury. Local renin-angiotensin system may play a role in the transdifferentiation of TEC through AT2 receptor and its JAK2 signal pathway.</p>