Application of MOOC in the teaching reform of human anatomy / 中华医学教育探索杂志

Objective:To explore the practical role of massive open online course (MOOC) mode based on the cultivation of creativity and thinking ability in clinical human anatomy teaching.Methods:Two classes of clinical medicine students of Batch 2019 were selected in the study, and one class was control group ( n=73), which adopted the traditional teaching mode of face-to-face teaching; the other class was research group ( n=79) and the MOOC mode based on the cultivation of creativity and thinking ability for teaching was adopted. After the teaching, the students' creativity, thinking ability, self-learning ability and learning interest were compared, and the mastery of knowledge (theoretical scores and anatomical operation assessment) and satisfaction with teaching were compared between the two groups. SPSS 19.0 was used for t test, chi-square test and rank sum test. Results:The scores of creativity, thinking ability, self-study ability and learning interest of the research group were significantly higher than those of the control group after the teaching ( P<0.05). The scores of theoretical knowledge[(91.41±6.28) points] and anatomical operation[(87.41±7.25) points] in the research group were significantly higher than those in the control group after the teaching[(85.24±7.36) points and (80.26±6.38) points] ( P<0.05). There was significant difference in the distribution of teaching satisfaction between the two groups ( P<0.05), and the total satisfaction rate for teaching of the research group (94.94%) was higher than that of the control group (83.56%). Conclusion:The MOOC mode based on the cultivation of creativity and thinking ability in clinical human anatomy teaching can significantly improve students' creativity, thinking ability and self-learning ability, improve their learning interest and mastery of human anatomy knowledge, and improve their satisfaction with teaching.

Clinical feature and variant analysis of a case with hereditary hypophosphatemic rickets with hypercalciuria / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical features and genetic basis for a patient with hereditary hypophosphatemic rickets with hypercalciuria(HHRH).@*METHODS@#Clinical data of the patient was collected. The patient was subjected to whole exome capture and next generation sequencing (NGS). Suspected variants were verified by Sanger sequencing.@*RESULTS@#The patient presented with hypophosphatemic rickets, short stature, hypercalciuria, and renal stones. NGS showed that he has carried compound heterozygous variants of the SLC34A3 gene, namely c.532_533delCA(p.Q178Vfs*6) and c.894_925+69del(splicing). His parents were asymptomatic heterozygous carriers of one of the variants. Based on ACMG guidelines, both variants were classified as pathogenic.@*CONCLUSION@#The compound heterozygous variants c.532_533delCA (p.Q178Vfs*6) and c.894_925+69del(splicing) of the SLC34A3 gene probably underlie the disease in this child. Above finding has enriched the variant spectrum for HHRH. Based on the results, prenatal diagnosis may be provided for the family.

Analysis of laboratory indexes and ultrasonic features of patients with Kawasaki disease shock syndrome / 中华实用儿科临床杂志

Objective@#To retrospectively analyze the laboratory findings and ultrasonographic features in acute phase of children suffering from Kawasaki disease (KD) with stable hemodynamics and Kawasaki disease shock syndrome (KDSS), so as to provide the evidence for early diagnosis, timely treatment and improvement of prognosis of KDSS.@*Methods@#Four hundred and eighteen patients with KD diagnosed at Shenzhen Children′s Hospital from November 2016 to May 2018 were selected, including 23 KDSS patients(KDSS group) and 395 cases with stable hemodynamic(KD without shock group). The clinical characteristics, laboratory index and ultrasonic examination data of the 2 groups were collected and compared for statistical conclusion.@*Results@#(1)The level of C-reaction protein(CRP)[166.20 mg/L (74.40 mg/L)], γ-glutamyl transpeptidase(γ-GT)[88.00 IU/L (126.00 IU/L)], creatine kinase isoenzyme(CKI)[1.78 μg/L (5.17 μg/L)], troponin(TP)[0.01 μg/L (0.39 μg/L)] in the KDSS group in acute phase were all higher than those in the KD without shock group[70.50 mg/L (54.30 mg/L), 40.00 IU/L (89.00 IU/L), 1.20 μg/L (0.85 μg/L), 0.01 μg/L (0.01 μg/L)], hemoglobin(Hb)[90.00 g/L (15.00 g/L)], ablumin [24.20 g/L (4.30 g/L)], serum sodium[130.90 mmol/L (5.60 mmol/L)] levels in the KDSS group were lower than those in the KD without shock group[107.00 g/L (14.00 g/L), 33.40 g/L (4.08 g/L), 136.10 mmol/L (3.25 mmol/L)], and the differences were statistically significant (all P<0.05). (2)The incidence rates of impaired left ventricular ejection fraction(LVEF)[<55%: 3 cases (13.03%) vs.8 cases (2.00%)], coronary artery abnormalities[left anterior descending branch(LAD) Z-score>2.5: 6 cases (26.09%) vs.35 cases (8.86%)]and valvular regurgitation[tricuspid regurgitation(TR)≥moderate: 3 cases (13.03%) vs.5 cases (1.26%)]in the KDSS group were higher than those in the KD without shock group, and the differences were statistically significant (all P<0.05). (3)Among acute phase in KDSS group, 9 cases (39.13%) had liver enlargement, 9 cases (39.13%) had peritoneal effusion, 3 cases (13.04%) had diffuse renal lesions, 3 cases (13.04%) had joint effusion (2 cases of knee joint effusion, 1 case of hip joint effusion), and 2 cases (8.70%) had enteritis.In the KD without shock group, only 3 cases (0.76%) had hepatomegaly and 2 cases (0.51%) had a small amount of knee effusion.@*Conclusions@#Laboratory findings of KDSS group showed higher level of CRP, CKI, TP as well as γ-GT than those in KD without shock group, who are more prone to suffer from hypohemoglobin, hyponatremia and hypoalbuminemia.Ultrasound examination showed that KDSS children were more prone to have heart or multiple-organ damage.Clinicians should raise their awareness to provide comprehensive assessment as well as timely and effective treatment.

Clinical features and mutational analysis of a case with Sensenbrenner syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the clinical characteristics of a patient with Sensenbrenner syndrome (also called cranioectodermal dysplasia type 3) caused by mutation of intraflagellar transport (IFT) 43 gene.</p><p><b>METHODS</b>The clinical data of the patient was retrospectively analyzed. The target genes was the patient were captured and subjected to next generation sequencing. Suspected mutations were verified through Sanger sequencing.</p><p><b>RESULTS</b>The patient, a-13 year-and-5-month-old girl, was admitted for anemia and renal dysfunction for 8 months. Clinically, she has featured short stature, short limbs, brachydactylia, tooth agenesis, and retinal dystrophy, high-degree myopia, and chronic renal failure. Gene sequencing showed that she has carried a homozygous c.1A>G (p.M1V) mutation of the IFT43 gene, for which both of her parents were heterozygous carriers.</p><p><b>CONCLUSION</b>c.1A>G (p.M1V) mutation of the C14ORF179/IFT43 gene is the cause for praecox chronic renal failure in children. Genetic testing can facilitate the diagnosis of this rare disorder. For affected families, prenatal diagnosis should be provided.</p>

IPEX with congenital ichthyosiform skin lesions:a case report and literature review / 临床儿科杂志

Objective To explore the clinical phenotype, treatment and prognosis of immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome, and to improve pediatricians' knowledge of this disease. Methods Clinical data of a case of IPEX with congenital ichthyosiform skin lesions were retrospectively analyzed, and related literatures China were reviewed. Results The 2-month-11-day old boy came to our hospital due to ichthyosiform skin lesions accompanied by blood oozing in the head and feet exudatation, with severe sepsis and gastrointestinal perforation. He was died of multiple organ failure. DNA sequencing of whole-genome exon group showed a hemizygous mutation of c.1150G> A, p.A384> T in FOXP3 gene. His mother was a heterozygous mutation carrier, while his father was normal. Conclusions In addition to typical symptoms including early-onset refractory diarrhea, multiple endocrine disease and growth retardation, IPEX should be considered also in infants with ichthyosiform rash and severe infection. Gene sequencing will help diagnose the disease.

The changes of regulatory B cell subset in children with type 1 diabetes mellilus / 中华内分泌代谢杂志

[Summary] The proportions of CD19+CD24 hi CD38 hi regulatory B cells ( Breg ) and CD4+CD25+Treg in peripheral blood of children with type 1 diabetes mellitus ( T1DM) were detected by flow cytometric analysis. The concentration of interleukin 10 (IL-10) protein was measured by ELISA. The mRNA expression levels of Foxp3, cytotoxic T lymphocyte-associated antigen-4 ( CTLA-4 ) , and glucocorticoid-induced tumor necrosis factor receptor ( GITR) in CD4+T cells and IL-10 in CD19+ B cells were evaluated using real-time PCR. The results showed that the proportions of CD19+CD24hiCD38hiBreg in peripheral blood, the mRNA and protein levels of IL-10 in B cells from patients with T1DM were lower than those from healthy controls (P<0. 05). The mRNA expression levels of Treg associated factors such as Foxp3, CTLA-4, and GITR were lower in CD4+ T cells from children with T1DM compared with the controls(P<0. 05). These results suggest that Breg cell deficiency and dysfunction might be one of the important factors causing cellular immune dysfunction in patients with T1DM.

Factors for differentiation affecting of neonatal regulatory T cells / 中华实用儿科临床杂志

Objective To investigate the possible factors for differentiation affecting of neonatal regulatory T cells(Treg). Methods Umbilical cord blood was collected from 200 newborns. Treg number was detected by DNA demethylation in the Foxp3 of Treg - cell - specific demethylatedregion(TSDR)based on high resolution melting anal-ysis(HRMA),concentrations of 7,8 - dihydroxy - 9,10 - epoxy - benzo(a)pyrene(BPDE - DNA)adducts and interleukin - 4( IL - 4)in the supernatants of cord blood by enzyme - linked immunosorbent assay( ELISA),and follow - up questionnaires were carried out till 1. 0 - 1. 5 years,for recurrent wheezing or stubborn eczema in infants and related information on parental history of atopic diseases. Results (1)In wheezing group[(0. 48 ± 0. 05)% ]and ec-zema group[(0. 76 ± 0. 05)% ],the number of Tregs was significantly lower compared with that of the asymptomatic group[(1. 14 ± 0. 08)% ](t = 2. 62,2. 83,all P ﹤ 0. 05);the number of Treg in parental history of atopic group was significantly lower than that of the non - atopic group(P ﹤ 0. 05);but the Treg numbers in the non - atopic group was still lower than that of the asymptomatic group(P ﹤ 0. 05).(2)The concentrations of BPDE - DNA adducts in the wheezing group[(236. 30 ± 6. 59)ng/ L]and the eczema group[(173. 40 ± 7. 38)ng/ L]were higher than those of the asymptomatic group[(111. 01 ± 3. 36)ng/ L](t = 10. 35,6. 53,all P ﹤ 0. 05),while BPDE - DNA adduct concen-trations in the atopic group with parental history of wheezing or eczema in infants were lower than those of the non -atopic group(P ﹤ 0. 05).(3)The concentrations of IL - 4 in the wheezing or eczema group in the supernatants of cord blood was higher than the asymptomatic group(P ﹤ 0. 05). Conclusions Neonatal genetic factors and BPDE - DNA adducts could affect Treg differentiation,which are probably the reasons for the formation of allergic diseases.

The role of activation of IL-6/STAT3 signaling in Th17/Tr imbalance of Kawasaki disease / 中华微生物学和免疫学杂志

Objective To investigate the role of IL-6/STAT3 signaling in Th17/Tr imbalance of Kawasaki disease(KD). Methods Forty-eight children with KD and eighteen age-matched healthy children were consented to participate in this study. Protein concentration of IL-6 in plasma was measured by ELISA. Transcriptional levels of IL-17A, IL-17F, RORγt, Foxp3, SOCS1 and SOCS3 were assessed by real-time PCR. The proportion of CD4+CD25+Foxp3+ regulatory T(Tr) cells and mean fluorescence intensity(MFI) for phosphorylated-STAT3(pSTAT3) protein in CD4+ T cells was analyzed by flow cytometry. A quantitative methylation specific PCR based on SYBR Green was used to evaluate methylation status of CpG islands in SOCS1 exon2, three potential bind sites for STAT3 in 5'-untraslated region(5'-UTR) of SOCS3 in CD4+ T cells. Results (1)Compared with healthy volunteers, plasma IL-6 concentration and MFI for pSTAT3 in CD4+ T cells were elevated significantly during acute phase of KD[IL-6:(54.02±20.58) pg/ml vs (8.72±2.06) pg/ml, P0.05). ConclusionAberrant activation of IL-6/STAT3 signaling caused by hypomethylation of SOCS1 and SOCS3 might be one contributing factor to unbalance of Th17/Tr in KD.

Investigation of the change of CD4~+ T cell subset from children with type 1 diabetes / 中华微生物学和免疫学杂志

Objective To investigate the changes of CD4~+ T cell subset in the role of immuno-pathogenesis of type 1 diabetes mellitus(T1DM). Methods Real-time PCR was used to evaluate the ex-pression levels of transcriptional factors (T-bet, GATA-3, Foxp3, ROR-γt), cytokines (IFN-γ, IL-4, IL-10, IL-17A) and negative regulatory factors (CTLA-4, GITR) mRNA from CD4~+ T cells. The proportions of Th1, Th2, Tr and Th17 cells in peripheral blood were detected by flow cytometric analysis. Plasma cyto-kine (IFN-γ, IL-4, TGF-β and IL-6) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) The proportions of Th1 cells in peripheral blood from children with T1DM were siguificanfly increased than that of healthy controls, and proportions of Th2 were decreased (P < 0.01). There were no significant differences between diabetic patients and healthy controls regarding the proportions of Tr cells and Th17 cells(P >0.05). (2) Transcription levels of T-bet and IFN-γ mRNA were significantly up-regulated, while GATA3 and IL-4 were significantly down-regulated in children with T1DM. The mRNA expression levels of Tr negativity regulatory factors such as IL-10, CTLA-4 and GITR were lower in CD4~+ T cells from children with TIDM compared with the controls(P <0.01). There were no statistically differences to be observed in mRNA expression levels of ROR-γt and IL-17A genes between two groups(P > 0.05).(3) In comparison with controls, serum concentrations of IFN-γ or IL-4 were remarkable increased or de-creased respectively (P < 0. 01), while TGF-β and IL-6 did not change in children with T1DM (P > 0.05). Conclusion The Th1/Th2 imbalance might be play an important role in immunopathogenesis of T1DM. Functional deficiency of Tr cell might further exacerbate Th1/Th2 imbalance and lead to disturbance of cellu-lar immune response.

The role of activation of toll-like receptors in immunological pathogenesis of Henoch-Schonlein purpura / 中华风湿病学杂志

ObjectiveTo investigate the role of signal transduction of TLRs in the Henoch-Schonlein purpura (HSP). Methods Reverse-transcription PCR (RT-PCR) and real-time PCR were used to evaluate the levels of TLRs(1～10), MyD88, TRAF6, TRIF, IFN-α, IFN-β, IL-6, IL-1β, TNF-α, IP-10, RANTES,iNOS, Blys/April mRNA expression in peripheral blood mononuclear cells and their expression levels were compared using t test., while the concentration of plasma cytokines such as Blys、IFN-α、IFN-β、IL-6、IL-1、TNF-α was measured by enzyme-linked immunosorbent assay(ELISA).Expression levels of those genes were compared using t test. Results①Compared with the control group, the expression levels of TLR1, TLR2,TLR6, TLR5, TLR3, TLR7, TLR9 mRNA were up-regulated significantly(P＜0.01), while no difference of TLR4 was detected (P＞0.05).②Transcription levels of MyDg8(2.47±1.06) vs(0.73±0.22), TRAF6 (2.54±0.72)×10-3vs(0.70±0.20)×10-3, TRIF(3.18±0.86)×10-3vs(0.93±0.35)×10-3 were significantly up-regulated in acute phase of HSP (P＜0.01).③The levels of IFN-α and IFN-β protein and mRNA were remarkable increased (P＜0.01).④ The expression of cytokine/chemotactic factor such as IL-6, IL-1β, IP-10, RANTES,iNOS was higher than that of the control group(p＜0.01), while TNF-αdid not change in children with HSP (P＞0.05). ⑤ It was detected that the expression of Blys/April was higher than that of the control group(P＜0.01). ConclusionExpressions of TLR1, TLR2, TLR6, TIR5, TLR3, TLR7, TLR9, MyD88, TRAF6, and TRIF are up-regulated during acute phase of HSP, suggesting that aberrant activation of TLRs triggered by microbes may be one of the initiating factors of immune aberrance in HSP. Over expression of cytokine/chemotactic factor or Blys/April owning to the aberrant activation of TLRs, may be correlated with immunological pathogenesis of HSP.

Influence of SOCS1 and SOCS3 hypomethylation on homeostasis of Th1/Th2 in Kawasaki disease / 中华风湿病学杂志

Objective To investigate the effect of SOCS1 and SOCS3 hypomethylation on homeostasis of Th1/Th2 in Kawasaki disease(KD). Methods Thirty-six children with KD and sixteen age-matched healthy children consented to participate in this study. Protein concentration of IL-6 in plasma was measured by ELISA. Transcriptional levels of SOCS1, SOCS3, T-bet, IFN-γ, GATA3 and IL-4 were assessed by realtime PCR. The proportion of Th1 and Th2 cells, and mean fluorescence intensity(MFI)for phosphorylated STAT3(pSTAT3)protein in CD4+ T cells was analyzed by flow cytometry. A quantitative methylation specific PCR based on SYBR Green was used to evaluate methylation status of CpG islands in SOCSl exon2, and three potential binding sites for STAT3 in 5'-untraslated region(5'-UTR)of SOCS3 in CD4+T cells. Comparisons between groups were performed with t-test. Results ①Compared with healthy volunteers, plasma IL-6 concentration[(51.8±16.3)pg/ml vs(8.6±2.0)pg/ml, respectively]and MFI for pSTAT3[(52±14)vs(10±4), respectively]in CD4+ T cells were elevated significantly during acute phase of KD(P＜0.05), and the two items in KD patients with coronary artery lesion(KD-CAL+)were found to be higher than those in KD patients without coronary artery lesion(KD-CAL-)[IL-6:(87.2±27.4)pg/ml vs(36.2±12.8)pg/ml, P＜0.05; pSTAT3 MFI:(75±15)vs(42±11), P＜0.05]. ② The proportions of Th1 and Th2 cells and transcription levels of Th-associating factors(T-bet, IFN-γ, GATA3 and IL-4)in CD4+ T cells increased significantly in acute KD(P＜0.05), while the rate of Thl div Th2 in KD patients was found to be lower than that in normal controls(P＜0.05). In addition, the proportions of Th1 and Th2 cells and expressions levels of Th-associating factors in KD-CAL+ group were higher than those in KD-CAL-group, as well as the rate of Thl div Th2 cells in KD -CAL+ group were lower than that in KD-CAL- group(P＜0.05). ③ The mRNA levels of SOCSl and SOCS3 in CD4+ T cells increased significantly during acute phase of KD(P＜0.05), while the two items in KDCAL+ group were lower than those in KD-CAL- group(P＜0.05). Furthermore, CpG islands in SOCSl exon2 and the third potential binding site for STAT3 in SOCS3 5'-UTR were hypomethylated in acute KD, while those in healthy volunteers were fully demethylated(P＜0.05). Demethylation levels of the two items mentioned above in the KD-CAL+ group were lower than those in the KD-CAL-group(P＜0.05). CpG islands in the other two binding sites for STAT3 in SOCS3 5'-UTR were fully demethylated among all the groups(P＞0.05).Conclusion Relative insufficiency of SOCS1 and SOCS3 expression caused by hypomethylation may be one contributing factor for the imbalance of Th1/Th2 in KD.

Investigation of the methylation status of Foxp3 gene during acute phase of Kawasaki disease / 中华微生物学和免疫学杂志

Objective To investigate the methylation status of Foxp3 gene and its roles in immunological pathogenesis of Kawasaki disease(KD). Methods Thirty children with KD and eighteen agematched healthy children consented to participate in this study. Quantitative methylation specific polymerase chain reaction(MSQP) was used to assess the methylation status of Foxp3 promoter and regulatory T cells specific demethylated region(TSDR) in CD4+ T cells. The proportion of CD4+ CD25 + Foxp3 + regulatory T (Tr) cells was analyzed by flow cytometry. Transcriptional levels of CD4+ CD25 + Foxp3 + Tr associating genes (Foxp3, CTLA4, GITR, LAG3 and CCR8 ) and Foxp3-dependent molecules (UBD and LGAIS3)were measured by real-time PCR. Results ( 1 ) Demethylation level of Foxp3 promoter in CD4 + T cells from patients with KD was lower significantly than that of health subjects( P ＜ 0.01 ), and increased significantly after treated with intravenous gamma globulin therapy(IVIG) (P ＜ 0.01 ). No difference of demethylation level of TSDR region was observed among all groups ( P ＞ 0. 05 ). ( 2 ) The proportion of CD4 + CD25 + Foxp3 + Tr in peripheral blood from patients with KD , as well as mRNA levels of Foxp3 gene in CD4+ T cells, was significantly lower than those of health subjects ( P ＜0. 01 ), and increases significantly after IVIG therapy (P ＜0.01 ). Significant positive correlations between demethylation level of Foxp3 promoter and the proportion of CD4 + CD25 + Foxp3 + Tr , or expression levels of Foxp3 in CD4 + T cells, were observed during acute phase of KD (CD4+CD25+ Foxp3+ Tr: r=0.76, P＜0. 01; Foxp3: r=0.89, P＜0. 01). (3)Transcription level of CD4+ CD25+ Foxp3+ Tr associating factors, such as CTLA4, GITR, LAG3 and CCR8, was significantly down-regulated in acute phase of KD(P＜0. 01 ), and up-regulated to some extent after treated with IVIG(P ＜0. 01 ). Expression levels of Foxp3-dependent molecules UBD and LGALS3 in CD4 + T cells decreased significantly during acute phase of KD (P ＜ 0.01 ), and basically recovered to the levels of health subjects( P ＜ 0.01 ). Conclusion Decrease of demethylation level of Fopx3 promoter is correlated with immune dysfunction in Kawasaki disease.

Significance and alteration of T help 17 cells in immune globulin-resistant gawasaki disease / 中华风湿病学杂志

Objective To investigate the alteration and significance of T help 17 cells (Th17), in patients with intravenous immune globulin-resistant Kawasaki disease (KD). Methods Forty-five children with KD (thirty-five as sensitive group and ten as the resistant group) and thirty age-matched healthy children were studied. Real-time PCR was used to evaluate the mRNA levels of IL-17A/F, ROR-γt. The proportion of Th17 in peripheral blood was detected by flow cytometric analysis. Th17-related plasma cytokine (including IL-17A and IL-6) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results ① Compared with healthy eontrols, transcription levels of IL-17A/F were significantly upregulated during acute phase of KD (P<0.01). ② Compared with sensitive group, the transcription levels of IL-17A/F, ROR-γt were significantly up-regulated during acute phase in immune globulin-resistant KD (P< 0.01), and down-regulated after treatment with intravenous immunoglobulin (IVIG) therapy in sensitive group (P<0.01), but there was no significant difference in the levels of expression in resistant group (P>0.05). There was significant difference in serum IL-17A,IL-6 before or after IVIG between resistant and nonresistant group (P<0.01). ③ Compared with the normal controls, serum IL-17A, IL-6 were markedly elevated in patients with KD. There was significant difference in serum IL-17A and IL-6 concentrations between resistant and sensitive group before IVIG treatment (P<0.01). The resistant group showed ignificantly high IL-17A, IL-6 after IVIG. There was no significant difference in the severity of decrease before and after IVIG in both groups, but it was found that serum IL-17A and IL-6 had a tendency to decrease after treatment with IVIG. ④ Methyiprednisolone might down-regulate inflammatory response through inhibiting the secretion of inflammatory IL-6 and IL-17A cytokines in acute stage of immunoglobulin-resistant KD. Treatment of immunoglobulin-resistant Kawasaki disease with IVMP resulted in faster resolution of fever, more rapid improvement in markers of inflammation. Conclusion Aberrant activation of Th17 cells may be one of the factors causing disturbed immunological function in immunoglobulin-resistant KD.