[Malfunction of autophagy in tibial growth plate chondrocytes causes increased apoptosis rate in chronic renal insufficiency rats].

Objective: To observe the effect of autophagy of tibial growth plate chondrocyte on apoptosis in chronic renal insufficiency (CRI) rats. Methods: Male 4-week-old SD rats were randomly divided into two groups: (1) Sham group: only the left ureter was exposed (n=10); (2) CRI group: the left ureter was ligated to cause CRI (n=10). The urine from all the rats was collected 6 weeks after the operation and the total protein content was measured. Then all the rats were sacrificed and the concentrations of creatinine and urea nitrogen in intracardiac blood were detected. The proximal tibia were fixed and decalcified to prepare histological sections, and the number of chondrocytes of column cells in the proliferative area of tibia growth plate was observed by saffron O staining. The expression rate of protein Light Chain-3, an autophagy marker of chondrocytes, was detected by immunofluorescence. The apoptosis rate of chondrocytes was detected by the method of TUNEL assay. The level of glycogenin-1, a glycogen formation marker of chondrocyte was detected by immunohistochemistry in chondrocytes. Results: The 24 h urine total protein was higher in CRI group [(163.5±11.3) mg vs (38.6±9.8) mg, t=25.620, P<0.001]. The levels of blood creatinine [(67.3±16.2) µmol/L vs (28.4±11.5) µmol/L, t=5.974, P<0.001] and urea nitrogen [(16.4±6.4) mmol/L vs (4.8±2.0) mmol/L, t=5.198, P<0.001] were higher in CRI group. The number of chondrocytes of column cells in the proliferating area of tibia growth plate was lower in CRI group (4.2±2.1 vs 9.1±3.8, t=3.109, P=0.006). The expression rate of LC-3 protein in chondrocytes of CRI group was lower [(27.2±12.6)% vs (51.4±18.2)%, t=3.457, P=0.003]. The level of glycogenin-1 of chondrocytes in CRI group increased significantly (6.1±2.5 vs 3.5±1.8, t=2.669, P=0.016). The apoptosis rate of chondrocytes in CRI group also increased [(17.2±4.8)% vs (5.1±3.4)%, t=6.505, P<0.001]. Conclusion: Malfunction of autophagy in tibial growth plate chondrocytes causes increased apoptosis rate in CRI rats, which might be caused by the failure of glycogen degradation in chondrocytes.

[Effects of remifentanil gradual withdrawal on remifentanil induced postoperative hyperalgesia].

Objective: To explore the effects of remifentanil gradual withdrawal on remifentanil induced postoperative hyperalgesia. Methods: Ninety patients from January to June 2018 undergoing elective laparoscopic hysterectomy under general anaesthesia at Ningbo NO.2 hospital, ASA Ⅰ or Ⅱ grade, aged 20-60, were enrolled in this study and randomly assigned to 3 groups (n=30): Group L (low dose remifentanil), Group H (high dose remifentanil) and Group G (high dose remifentanil with gradual withdrawal). Mechanical pain thresholds (MPT), visual analogue scale (VAS) and additional analgesics were recorded at 6 and 24 hours after the operation. Results: There was no significant difference among the VAS and additional analgesics in three groups at 6 and 24 hours after operation (P>0.05). There was no significant difference among the MPT in three groups before operation (P>0.05). The MPT of group L, group H and group G were (49.8±12.2), (35.5±13.0) and (48.6±11.4) g at 6 hours after surgery, and (51.4±14.3), (36.9±11.1) and (48.8±11.5) g at 24 hours after surgery, respectively, with statistically significant differences (F=12.6, 11.668, both P<0.01). The MPT of group H at 6 h and 24 h after surgery was lower than that of group L (all P<0.01), while the MPT of group G at 6 h and 24 h after surgery was significantly higher than that of group H (all P<0.01). No correlation was observed between MPT and VAS scores or additional analgesics at 6 and 24 hours postoperatively (P>0.05). Conclusion: Remifentanil gradual withdrawal significantly alleviated intraoperative administration of remifentanil induced postoperative hyperalgesia in laparoscopic hysterectomy patients.

[Effect of low expression of PTHrp receptor on proliferation of tibial growth plate chondrocytes in chronic renal insufficiency young rats].

Objective: To observe the effect of parathyroid hormone-related protein (PTHrp) receptor on the proliferation of tibial growth plate chondrocytes in chronic renal insufficiency (CRI) young rats. Methods: Two-week-old male SD rats were randomly divided into two groups: (1) Sham group (n=6), only left ureter was exposed; (2) CRI group(n=6), left ureter was ligated to induce chronic renal insufficiency. Rats were sacrificed 2 weeks after operation and the blood concentration of PTHrp was detected by intracardiac blood sampling. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. The level of PTHrp receptor in chondrocytes was observed by immunohistochemistry and quantitative analysis was completed by Western blot. The proliferation rate of chondrocytes from two groups at 24 h was detected by using 5-ethynyl-2'-deoxyuridine (EDU) technique. Three types of PTHrp receptor mRNA plasmids (overexpressed, empty vector and knockdown) were used to treat the chondrocytes from CRI group. The mRNA and protein levels of PTHrp receptor were detected after 24 h and 48 h intervention, respectively. The chondrocyte proliferation rate at 24 h was detected by EDU. Results: Blood concentration of PTHrp in CRI group was higher than that in Sham group [(1.36±0.42) ng/L vs (0.77±0.21) ng/L, t=3.913, P=0.001]. The results of Western blot showed that the level of PTHrp receptor in growth plate chondrocytes from CRI group decreased (0.15±0.07 vs 0.41±0.13, t=5.569, P<0.001). Chondrocyte proliferation rate of CRI group was lower than that in Sham group at 24 h [(11.3±3.1)% vs (24.6±5.7)%, t=6.482, P<0.001]. The mRNA and protein levels of PTHrp receptor increased in chondrocytes of CRI group after intervention with overexpressed plasmid. The chondrocyte proliferation rate increased at 24 h. On the contrary, the mRNA and protein levels of PTHrp receptor decreased afer intervention with knockdown plasmid, and the chondrocyte proliferation rate also decreased [overexpression: (22.8±6.5)%, empty carrier: (10.2±4.3)%, knockdown: (5.6±2.1)%, F=29.840, P<0.001]. Conclusion: Increased PTHrp concentration in the blood of CRI young rats leads to decreased PTHrp receptors in growth plate chondrocytes, which results in decreasing PTHrp activity and proliferation rate of chondrocyte.

[Characteristics of ERG, Fli-1, CD34, CD31 and Fâ §RAg expression in hepatic malignant vascular tumors].

Objective: To investigate the expression of ERG, Fli-1, CD34, CD31 and factor Ⅷ-related antigen(FⅧRAg) in hepatic malignant vascular tumors. Methods: A retrospective analysis was conducted on 63 cases of primary hepatic malignant vascular tumors and 31 cases of hepatic other malignant spindle cell tumors collected during January 1986 to January 2014. EnVision method was used to detect the expression of ERG, Fli-1, CD34, CD31, FⅧRAg. Results: Sixty-three cases of malignant vascular tumors, including 24 cases of angiosarcoma, 38 cases of epithelioid hemangioendothelioma and 1 case of hepatic Kaposi's sarcoma. All of the cases were positive for ERG(100.0%, 63/63). Positive rate of Fli-1, CD34, CD31, FⅧRAg was 96.8% (61/63), 87.3% (55/63), 81.0% (51/63) and 41.3% (26/63), respectively. In other hepatic malignant spindle cell tumors, the positive rate of ERG, Fli-1, CD34, CD31 and FⅧRAg was 3.2% (1/31), 19.4% (6/31), 19.4% (6/31), 9.7%(3/31) and 3.2%(1/31), respectively.The sensitivity of ERG, Fli-1, CD34, CD31, FⅧRAg was 100.0%, 96.8%, 87.3%, 81.0% and 41.3%, respectively.The specificity was 96.8%, 80.6%, 80.6%, 90.3% and 96.8%, respectively. Conclusion: ERG is a more sensitive and specific diagnostic marker for hepatic malignant vascular tumors in comparison to Fli-1, CD34, CD31 and FⅧRAg.

[Phosphatase and tensin homolog deleted on chromosome ten/phosphatidyl Inositol 3-kinase/vascular endothelial growth factor signaling pathway changes in the rabbit Kawasaki disease model].

OBJECTIVE: To observe the changes of phosphatase and tensin homolog deleted on chromosome ten(PTEN)/ phosphatidyl Inositol 3-kinase(PI3K)/ vascular endothelial growth factor(VEGF)signaling pathway in a rabbit Kawasaki disease model. METHODS: Model of Kawasaki disease was established in weanling Japanese big-eared rabbits with 10% bovine serum venous injection (2.5 ml/kg, 2 times, and 2 week's interval) through the ear. Twenty four rabbits were divided into 4 groups: control group (without injection of 10% bovine serum albumin, six rabbits); 1 day group (sacrificed a the second day after the establishment of Kawasaki disease models, six rabbits); 7 day group (sacrificed at the seventh day after establishment of Kawasaki disease model, six rabbits); 30 day group (sacrificed at the thirtieth day after establishment of Kawasaki disease model, six rabbits). Pathological analysis was performed on coronary artery tissue samples. The express of PTEN and PI3K were detected by immunohistochemistry. The levels of VEGF and CK were also examined with ELISA and white blood cells were counted. RESULTS: (1) Coronary artery of model groups was thinner, distorted and had enlarged lumen. (2) PTEN expression in 1 d group, 7 d group and 30 d group were 58.5 ± 12.9, 73.2±9.9 and 109.6 ± 24.4, respectively, significantly higher than in the control group (25.5 ± 6.9, P<0.01 or 0.05). (3) The express of PI3K was significantly upregulated in 1 d group(57.2±11.1)and 7 d group(39.9±4.8) compared to control group(19.1±3.5, P<0.01 or 0.05). The expression level of PI3K in 30 d group was 18.8 ± 7.5, which was similar as control group (P>0.05) and significantly lower than 1 d and 7 d group (both P<0.05). (4) Similarly, the level of VEGF in 1 d group, 7 d group ((89.1 ± 15.5) ng/L, (76.9±9.9) ng/L) were significantly higher while it was significantly lower in 30 d group ((19.8 ± 4.4)ng/L) compared with the control group ((33.9 ± 6.7) ng/L, P<0.01 or 0.05). The level of VEGF in 7 d group was significantly lower than in 1 d group (P<0.05) and the level of VEGF was significantly lower in 30 d group than in 7 d group (P<0.01). (5)Creatine kinase levels were significantly higher in 30 d group than in control group (P<0.05) and there were no significantly different between control group, 1 d group and 7 d group(all P>0.05). (6)White blood cell count were significantly higher in 1 d group, 7 d group and 30 d group than in control group (all P<0.01). CONCLUSION: The level of PTEN/PI3K/VEGF signaling pathway change after establishment of rabbit Kawasaki disease model and the signaling pathway might be involved in this model.

Immature dendritic cells expressing indoleamine 2,3-dioxygenase suppress ovalbumin-induced allergic airway inflammation in mice.

BACKGROUND: Proliferation of activated CD4+ T lymphocytes is inhibited by indoleamine 2,3-dioxygenase (IDO). OBJECTIVE: We undertook the present study to test the hypothesis that IDO-expressing immature DCs (imDCs) can restore immune tolerance in mice suffering from allergic airway inflammation. METHODS: imDCs were generated from murine bone marrow cells using granulocyte-macrophage colony-stimulating factor.The imDCs were subsequently transfected with an IDO expression vector (pEGFP-N1-IDO). Surface marker expression, including CD11c, MHCII, CD80, and CD86, was analyzed using flow cytometry. IDO-expressing imDCs were injected into the trachea of ovalbumin (OVA)-sensitized mice, and lung histopathology and cytokine expression in bronchoalveolar lavage fluid were assessed. The splenic CD4+ T cells of OVA-sensitized mice were isolated and co-cultured with pEGFP-N1-IDO-expressing imDCs, and apoptosis of CD4+ T cells was detected using the terminal deoxynucleotidyl transferase dUTP nick end labeling assay. RESULTS: Expression of IDO in imDCs did not alter cell surface molecule expression. We observed marked lung inflammation, elevated total cell and eosinophil count, and altered cytokine levels in OVA-sensitized mice. These parameters improved upon inoculation with IDO-expressing imDCs. Co-culture with IDO-expressing imDCs also induced apoptosis, inhibited IL-4 and IL-5 expression, and upregulated IFN-gamma expression in CD4+ T cells. CONCLUSIONS: IDO-expressing imDCs induced T(H)2 cell apoptosis and reduced T(H)2 cell activation and allergic airway inflammation in OVA-sensitized mice. Thus, upregulation of IDO expression may provide a novel immunointervention strategy for asthma treatment.

[Loss of autophagy in condylar chondrocytes causes increased apoptosis rate in temporomandibular joint osteoarthritis of rats].

Objective: To observe the effect of autophagy of condylar chondrocytes on apoptosis in temporomandibular joint osteoarthritis (TMJOA) of rats. Methods: Fourty male 2-month-old SPF SD rats were equally divided into sham group (n=20) and experimental group (n=20). UAC metal prosthesis was cemented to the left incisors of maxilla and mandible of the rats in experimental group rats. After 8 weeks, all rats were sacrificed and the temporomandibular joint was taken. Two groups of rat condylar chondrocytes were extracted and cultured in vitro to the third generation. Immunofluorescence technique was used to detect the levels of collagen Ⅱ and matrix metalloproteinase-13 (MMP-13) in chondrocytes. The level of light chain-3 (LC-3), an autophagy marker of chondrocytes, was detected. Immunohistochemical technique was used to detect the level glycogenin-1, a glycogen formation marker of chondrocyte, was detected. The level of caspase-3, an apoptosis marker of chondrocyte, was also detected. Tunel technique was used to detect the apoptosis rate of the two groups at 72 h. Cracking cell extraction of total protein, Western-blotting (WB) technology to detect the levels of collagen Ⅱ, MMP -13, LC-3, glycogenin-1, caspase-3 and make gray analysis. Results: Compared with sham group, the level of collagen Ⅱ decreased, MMP-13 increased, LC-3 decreased, glycogenin-1 increased and caspase-3 increased in experimental group. The apoptosis rate of chondrocytes in experimentaal group [ (17.3±4.4) %] at 72h was higher than that in control group [ (5.6±2.1) %](t=10.732, P<0.001) .WB bands gray statistical results show that the level of collagen Ⅱ in chondrocytes of experimental group (0.43±0.21) was lower than that of control group (0.71±0.26) (t=2.409, P=0.043) , the level of MMP-13 in chondrocytes of experimental group (0.73±0.31) was higher than that of control group (0.24±0.10) (t=3.364, P=0.010) , the level of LC-3 in chondrocytes of experimental group (0.09±0.04) was lower than that of control group (0.39±0.18) (t=3.638, P=0.007) , the level of glycogenin-1 in chondrocytes of experimental group (0.68±0.30) was higher than that of control group (0.29±0.17) (t=2.529, P=0.035) , the level of caspase-3 in chondrocytes of experimental group (0.19±0.08) was higher than that of control group (0.05±0.02) (t=3.796, P=0.005) . Conclusions: The level of autophagy of condylar chondrocytes in temporomandibular joint of rats decreased, glycogen accumulation increased, the rate of chondrocyte apoptosis increased, and the number of chondrocytes decreased, resulting in degeneration of condylar cartilage tissue.

The efficacy observation of ulinastatin combined with creatine phosphate sodium in pediatric viral myocarditis.

OBJECTIVE: This study aimed to compare the efficacy and safety of ulinastatin combined with creatine phosphate sodium and ribavirin combined with creatine phosphate sodium in the treatment of pediatric viral myocarditis. PATIENTS AND METHODS: 155 children with viral myocarditis in the Xuzhou Children's Hospital, were retrospectively analyzed. 80 of them received ulinastatin combined with creatine phosphate sodium, and were regarded as observation group; other 75 patients received ribavirin combined with creatine phosphate sodium and were regarded as the control group. The therapeutic efficacy of the two groups was observed, the improved condition of myocardial enzyme indicator and myocardial troponin I (cTn I) in the two groups were compared before and after the treatment. RESULTS: The total effective rates of the patients in the observation group and the control group were 93.75% and 76.00%, respectively. The clinical efficacy of the observation group was better than that of the control group (p<0.05). The electrocardiogram improvement condition of the observation group was better than that of the control group (p<0.05); after the treatment, the expression levels of lactate dehydrogenase (LDH), aspartate aminotransferase (AST), Creatine Kinase (CK-MB), and cTn I in the observation group were (313.37±9.42) U/L, (29.38±4.97) U/L, (23.67±2.89) U/L, (0.12±0.02) µg/L, respectively. The expression levels of LDH, AST, CK-MB, and cTn I in the control group were (322.43±12.32) U/L, (33.43±5.14) U/L, (26.22±3.37) U/L, (0.24±0.04) µg/L. The levels of myocardial enzyme and cTn I in the observation group and the control group after the treatment were lower than that before the treatment (p<0.05), while the level of myocardial enzyme and cTn I in the observation group after the treatment was significantly lower than that in the control group (p<0.05). CONCLUSIONS: The results indicated that, compared with ribavirin combined with creatine phosphate sodium, ulinastatin combined with creatine phosphate sodium had better clinical efficacy in the treatment of pediatric viral myocarditis. It could significantly improve the level of myocardial enzyme indicator and cTn I, and had certain clinical and promotional values and application values.

Analysis of the correlation between the neutrophil-lymphocyte ratio in peripheral blood and perioperative myocardial damage in pediatric patients with frequent ventricular premature beat.

OBJECTIVE: The objective of the present study was to investigate the relationship between the neutrophil-lymphocyte ratio (NLR) in peripheral blood and myocardial damage in pediatric patients with frequent ventricular premature beat (FVPB), and provide a reference for myocardial preservation in these patients. PATIENTS AND METHODS: A total of 212 pediatric patients who were treated in the Department of Cardiology, Xuzhou Children's Hospital between December 2014 and March 2016 for FVPB, were selected. The results of routine blood exam, and levels of cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB) in patients after the onset of FVPB were analyzed, and NLR was calculated. According to NLR levels, patients were divided into four groups using quartiles. RESULTS: With increases of NLR, the proportion of patients with a history of hypertension and ejection fraction < 50% increased gradually, and white blood cells (WBCs), the peaks of CK-MB and cTnI, and serum creatinine levels were significantly increased (p<0.05, p<0.01). There were no significant differences (p>0.05) in age, sex, body mass index, serum creatinine before treatment, fasting blood glucose, TG, TC, LDL-C, and HDL-C among the four groups. Multiple stepwise regression analysis showed that for patients with FVPB, NLR in peripheral blood was positively correlated with the peak of cTnI (r=0.538, p<0.05). NLR was positively correlated with levels of WBCs (r=0.661, p<0.05) and there was no correlation between NLR and history of hypertension, ejection fraction, and the laboratory results of creatinine peak and CK-MB. The differences were not statistically significant (p>0.05). However, the peak of cTnI was positively correlated with the levels of WBCs (r=0.189, p=0.003). CONCLUSIONS: NLR and WBCs in patients with FVPB are positively related to the peak of cTnI. NLR may serve as an excellent marker that reflects myocardial damage in pediatric patients with FVPB.

Heart rate variability in children with myocarditis presenting with ventricular arrhythmias.

OBJECTIVE: We aimed to investigate the heart rate variability in children with myocarditis presenting with ventricular arrhythmias. PATIENTS AND METHODS: The study compared the characteristics of heart rate variability (HRV) among 67 children with viral myocarditis (VMC), presenting with (n=35) and without (n=32) ventricular arrhythmias and a control group of 30 healthy children. RESULTS: Compared with the control group, the HRV time-domain indicators of children with VMC were significantly lower (p<0.05); also, the indicators of children with ventricular arrhythmias were significantly lower than those of children without ventricular arrhythmias (p<0.05). Equally, during both the lucid and sleep periods, the time-domain indicators of HRV were significantly lower in patients with VMC and arrhythmias than in either the control group (p<0.05) or the group with VMC but no ventricular arrhythmias (p<0.05). CONCLUSIONS: We conclude that the HRV of children with VMC probably decreased because of impaired vagal nerve function, with ventricular arrhythmias developing only when the decrease was most significant. Thus, HRV can be a useful predictive indicator for ventricular arrhythmias in children with VMC.

Transient receptor potential melastatin 4 cation channel in pediatric heart block.

OBJECTIVE: Progressive cardiac conduction disease (PCCD) is a common pediatric heart conduction disorder. It is an autosomal inheritance of rare mutations, which leads to familial cases of PCCD. In these cases, the His-Purkinje system's conductive capacity is progressively deranged, involving either right or left bundle branch block. Also, QRS complexes display widening is an important characteristic that culminates in complete AV block, syncope, and sudden death. Mutations in TRPM4 gene that encodes for transient receptor potential melastatin 4 have recently been reported to cause familial cases of PCCD and heart block. TRPM4 conducts a Ca2+-activated non-selective monovalent cationic current leading to a negative plasma membrane potential. TRPM4 channels let Na+ ion influx, causing membrane depolarization, whereas, at positive membrane potentials, TRPM4 channels repolarize the membrane by facilitating K+ ion efflux from the cell. TRPM4 protein contains many regulatory motifs that confer voltage dependence, ATP/ADP sensitivity, and Ca2+ responsiveness. Mutational studies revealed the significance of the two-calmodulin binding sites at the N-terminus of for Ca2+ dependent activation of this channel. Mutations that reduce deSUMOylation increase the steady-state levels of active TRPM4 channels on the membrane without alteration of its sensitivity to Ca2+ or ATP or its voltage dependence of activation. Increased TRPM4 function interferes with cardiac conduction and eventually contributes to heart block. Both gain and loss of function mutations of TRPM4 are implicated in the cardiac block. Currently, the major therapeutic management of cardiac block due to TRPM4 mutations is implantation of a pacemaker to reinstate normal current propagation through AV node.

Relevant researches on chronic viral myocarditis (CVMC) in children, complicated with arrhythmia and thyroid hormone level.

OBJECTIVE: To analyze the correlation between chronic viral myocarditis (CVMC) in children, complicated with arrhythmia and thyroid hormone level. PATIENTS AND METHODS: 60 patients with CVMC complicated with arrhythmia were continuously selected (course of disease > 3 months) and they are were diagnosed with arrhythmia by the routine 18-lead electrocardiogram and 24-hour Holter; the average follow-up time is about 2 years, during which the left ventricular end-diastolic diameter (LVEDd), left ventricular ejection fraction (LVEF), the occurrence rate of malignant arrhythmia events, immune state of T cell and thyroid hormone level (FT3, FT4, TSH, TGAb and TPOAb) were compared. RESULTS: Among the selected 60 patients, 18 patients (30.0%) who were suffering from malignant arrhythmia have been taken as the observation group. When compared with the control group, the standard deviation normal to normal intervals (SDNN), LVEF, CD4 and CD4/CD8 were reduced and LVEDd and CD8 were increased in the observation group; the difference has statistical significance (p < 0.05). When compared with the control group, FT3 and FT4 are significantly reduced and TSH, thyroglobulin antibody TGAB and thyroid peroxidase antibodies (TPOAb) are significantly increased; the difference has statistical significance (p < 0.05). According to the logistic regression analysis, we can conclude that: SDNN, FT3, FT4, TSH, TGAb and TPOAb are the independent risk factors of malignant arrhythmia (p < 0.05). CONCLUSIONS: Thyroid hormones and antibody level are helpful to the prognosis of malignant arrhythmia resulting from children chronic VMC complicated with arrhythmia complications.

Research on the relativity between gene polymorphism and children cardiac insufficiency.

OBJECTIVE: We analyzed the relationship between Mink-S27 gene polymorphism and children with cardiac insufficiency. PATIENTS AND METHODS: From April 2013 to April 2015, we enrolled 73 cases of children with cardiac insufficiency for this study, and all 73 were placed in the observation group. 76 normal cases were selected for the control group. Restriction fragment length polymorphism (RFLP) was used to make polymorphism analysis of the Mink-S27. RESULTS: Our results showed no significant differences in Mink-S27 genotype and allele distribution in both observation and control groups (p>0.05). In lesion samples collected from children with cardiac insufficiency, we detected significant difference in AA, CC genotype frequency and allele frequency between the observation group and the control group (p< 0.05) (X2 = 15.43, p<0.05; X2  = 16.27, p<0.05). Further studies on samples obtained from both groups revealed certain differences of AA, CC, AC genotype frequency and allele frequency in the observation group. The proportion of homozygote (AA, CC) in children with severe cardiac insufficiency was relatively high. CONCLUSIONS: GNAS2 gene polymorphism was associated with the prevalence of cardiac insufficiency in children. And also the patients' condition was correlated to the frequency of different genotypes and alleles.

Correlation between NFATC1 gene polymorphisms and congenital heart disease in children.

OBJECTIVE: To analyze the links between NFATC1 gene polymorphism and congenital heart disease in children. PATIENTS AND METHODS: In the present study, we selected 85 children patients with congenital heart disease who were hospitalized from February 2013 to February 2015 as research subjects (observation group), and 92 healthy subjects as control group. Restriction fragment length polymorphism (RFLP) was used for analysis of NFATC1 gene in samples from each group. RESULTS: The distribution of NFATC1 genotype and allele between the observation group (children with congenital heart disease) and the control group showed no significant difference (p >0.05), but AA, GG genotypes, and allele frequency between pathological samples of children with congenital heart disease and the control group displayed significant difference (p <0.05) (X2 = 16.04, p <0.05; X2 = 16.29, p <0.05). Further analyses showed that AA, GG, AG genotype and allele frequency among children with congenital heart disease in observation group also showed a difference, i.e., homozygote (AA, GG) ratio in children with severe congenital heart disease is relatively high. CONCLUSIONS: There is a correlation between NFATC1 genes and the incidence of congenital heart disease in children, and a correlation between different genotypes and allele frequency and the incidence of the disease.

Macrophage activation syndrome as a complication of juvenile rheumatoid arthritis.

OBJECTIVE: Juvenile rheumatoid arthritis (JRA), also known as juvenile idiopathic arthritis (JIA), is a rare autoimmune joint disorder of children. The concrete causes for the prevalence of the above pathological state are still unknown. In other words, it is an arthritis affecting mainly children and adolescents. Clinically, it has 3 different clinical subtypes. JRA patients are often noticed with some confirmed symptoms including coagulopathy, disseminated intravascular coagulation (DIC) with hepatosplenomegaly, fall in erythrocyte sedimentation rate and higher levels of liver enzymes leading to a life-threatening outcome. The above complications of JRA are recognized as a macrophage activation syndrome (MAS), which is similar to hemophagocytic lymphohistiocytosis (HLH). Pathogenesis of JRA manly involves deregulation of immunological processes with excessive and persistent activation of antigen presenting cells and T-lymphocytes. Further, abnormalities in the functioning of NK cells are often observed in JIA cases. Also, 40% of patients with these abnormalities are habitually associated with perforin gene mutations. Today, MAS remains a clinical and diagnostic challenge. RESULTS: The diagnosis of MAS is mainly based on clinical grounds. However, laboratory evidence of macrophages in the bone marrow performing phagocytosis of variable hematopoietic cells also help in diagnosis. For confirmation of MAS, there must be present either of two clinical or laboratory criteria. Further, laboratory criteria often appear late and are unable to diagnose the complication right at the beginning stage. Important laboratory findings in macrophage activation syndrome associated with JIA include hypertriglyceridemia, anemia, low erythrocyte sedimentation rate, elevated alanine aminotransferase level, higher than normal bilirubin levels, presence of fibrin degradation products, high lactate dehydrogenase level, low sodium, low albumin, and hyperferritinemia. CONCLUSIONS: MAS is a confirmed life threatening complication of patients with JIA. Further, an early diagnosis and treatment of MAS could be a life-saving mode for this syndrome.

Correlation analysis between GNAS2 gene polymorphisms in children with arrhythmia.

OBJECTIVE: The objective of the present study was to analyze the correlation between GNAS2 gene polymorphisms and children with arrhythmia. PATIENTS AND METHODS: We followed 89 children with arrhythmia treated in our hospital from April 2013 to April 2015, comprising the observation group, while 92 healthy subjects were taken as the control group. We analyzed polymorphisms of the GNAS2 gene from both groups by restriction fragment length polymorphism (RFLP). RESULTS: There were no significant differences (p>0.05) in GNAS2 genotype or allelic distribution between the observation group and control group. However, there were significant differences in the gene frequency of homozygotes (TT and GG) and alleles between the control group and samples from children with arrhythmia (p<0.05) (X2=16.57, p<0.05; X2=17.48, p<0.05). Further study of samples from both groups indicate that gene frequency of homozygous (TT and GG) or heterozygous (TG) genes and alleles also had differences, indicating that homozygotes (TT and GG) are in relatively high proportion among patients with severe cases of arrhythmia. CONCLUSIONS: Morbidity in patients with arrhythmia is related to GNAS2 gene polymorphisms. Differences in genotype and allele frequency are related to the severity of illness.

The application of critical pathway management to resident doctor's standardized training in pediatrics.

OBJECTIVE: To further guide the standardized training work of the resident doctor and apply critical pathway management to resident doctor's standardized training. MATERIALS AND METHODS: The traditional training contents were adjusted. The main research content of evaluating the results of standardized training was designed, and the research method was adopted by the standardized training of the resident doctor. RESULTS: The resident doctors of the group with the new teaching method demonstrated a high participation rate. Because of the constraint of the standardized training, trainers would pay more attention to their training. CONCLUSIONS: The clinical pathway polishes up the ability of resident doctors, further guiding the standardized training work of a resident doctor.

Endothelial microparticles induce vascular endothelial cell injury in children with Kawasaki disease.

OBJECTIVE: To explore the role of microparticles produced by endothelial cells in the injury of vascular endothelial cells. MATERIALS AND METHODS: We stimulated human umbilical vein endothelial cells (HUVEC) with TNF-α in vitro, analyzed the change of cellular morphology, and measured EMP level in the supernatant with a flow cytometer. Then, we evaluated the corresponding clinical indicators and the role of EMP in endothelial injury. RESULTS: The endothelial cellular morphology underwent significant changes, and a large number of microparticles were secreted. In turn, these microparticles blocked cell cycle and induced apoptosis. CONCLUSIONS: The microparticles produced by endothelial cells play an important role in the injury of vascular endothelial cells.

124 cases of clinical analysis of children with viral myocarditis.

OBJECTIVE: A study on clinical characteristics, diagnostic methods of children's viral myocarditis (VMC) and analysis of the effect of VMC is carried out. PATIENTS AND METHODS: A retrospective analysis of the clinical data of 124 cases of children with VMC from January 2008 to March 2011 in Xuzhou Children's Hospital is conducted. RESULTS: The age of onset of 1 VMC is less than 1 year old with the majority, fever, precordial pain, and fatigue as the main manifestation, auxiliary examination of abnormal electrocardiogram (ECG) in 110 cases, accounting for 88.71%, the positive rate of dynamic ECG is 92.31%, and abnormal rate of myocardial enzyme examination is 96.77%. The effective rate of treatment is 94.35%, and the effective rate of treatment of creatine phosphate sodium is 100%. CONCLUSIONS: For childhood, especially 1 year old children with viral infection in clinical, attention should be paid to the VMC performance, suspected children should be timely ECG, dynamic ECG and myocardial enzyme examined with diagnosis and early treatment. The myocardial protection of creatine phosphate sodium has definite therapeutic effect on VMC of children.