[Analysis of the Characteristics of the Outcomes of Job-Transfer Subspecialty Training Program in Pediatrics in Sichuan].

Objective: To discuss the characteristics of physician trainee outcomes after completion of the job-transfer subspecialty training in pediatrics, a program designed to increase the number of pediatricians, in Sichuan Province and to provide countermeasures for alleviating the shortage of pediatricians. Methods: We collected with questionnaire surveys information on changes in the workload and salaries experienced by physicians who completed the job-transfer subspecialty training program in pediatrics between February 2017 and May 2020 in Sichuan Province. Then, we compared the characteristics of physicians who successful became pediatricians and those who did no. Results: A total of 208 physicians completed the job-transfer subspecialty training program in pediatrics. Among them, 178, accounting for 85.6%, completed the questionnaire survey, and 120, accounting for 67.4%, had a background in other subspecialties than pediatrics. The majority (>90%) of physicians who participated in the training program came from secondary or lower levels of hospitals from the cities and prefectures all over Sichuan Province. In this study, we found that the rate of successful job transfer from being a physician to being a pediatrician in Sichuan Province in the past four years was 85.0% (102/120), with the year-by-year results being 88.2% (15/17) in 2017, 72.7% (16/22) in 2018, 86.7% (39/45) in 2019, and 94.% (32/34) in 2020. There was no significant difference between physicians who had successful job transfer and became pediatricians and those who failed to do so in terms of gender, age, hospital level, specialization prior to the job transfer, whether or not the hospital had a pediatrics department, amount of support for the pediatrics department, whether or not the physician was working at a new hospital after the job transfer, salaries, and changes of responsibilities during COVID-19 (all P>0.05). There was significant difference in the change of workload after completion of the training program between physicians who had successful job transfer and became pediatricians and those who failed to do so ( χ 2=9.037, P=0.003), and 78.4% of the trainees stated that their workload had increased after the job transfer. There was a moderate correlation between successful job transfer and changes in workload after the transfer (|Phi[ψ] |=0.729). Conclusions: The policy of government-supported job-transfer subspecialty training in pediatrics has played an active and important role in the swift resolution of the shortage of pediatricians. However, finding the root cause of and addressing the problem of the overwhelming workload of pediatricians remain challenging issues to be resolved.

Crumbs homolog 2 mutation in two siblings with steroid-resistant nephrotic syndrome: Two case reports.

BACKGROUND: Crumbs homolog 2 (CRB2) is a recently discovered gene that is closely related to the maintenance of normal polarity in podocytes; mutations can directly lead to steroid-resistant nephrotic syndrome (SRNS). However, the characteristics of nephrotic syndrome (NS) caused by CRB2 mutations have not been described. CASE SUMMARY: We report a novel compound heterozygous mutation of the CRB2 gene in two siblings with SRNS. The two siblings had edema, proteinuria, hypoproteinemia and hyperlipidemia. Both their father and mother had normal phenotypes (no history of NS). Whole exon sequencing (WES) of the family showed a novel compound heterozygous mutation, c.2290 (exon 8) C > T and c.3613 (exon 12) G > A. Glucocorticoid therapy (methylprednisolone pulse therapy or oral prednisone) and immunosuppressive agents (tacrolimus) had no effect. During a 3-year follow-up after genetic diagnosis by WES, proteinuria persisted, but the patient was healthy. CONCLUSION: CRB2 mutations related to SRNS often occur in exons 7, 10, and 12. Clinical manifestations of SRNS caused by CRB2 mutations are often less severe than in other forms of SRNS.

Tuning of nanofocused vector vortex beam of metallic granary-shaped nanotip with spin-dependent dielectric helical cone.

We present the combined configuration of dielectric helical cone and metallic granary-shaped nanotip to produce three -dimensional vector vortex nanofocused optical field. The intensity and phase of the electric fields, and Povnting vector of the optical field generated by the combined configuration with linearly polarized illumination are studied with three-dimensional finite difference time-domain method. The localized vector electric field near the apex of the metallic granary-shaped nanotip is strongly depended on the chirality of the dielectric helical cone and the bottom radius of the metallic granary-shaped nanotip. The localized vector electric field is wavelength selective with the maximum intensity enhancement up to 104 times and minimum size of about 900 nm2, and the maximum radial electric field rotates 67.0° along z axis. This indicates the vector vortex beam generated by the combined configuration can be applied in nanofabrication, nano-sensing and nano-manipulation.

Children with Steroid-resistant Nephrotic Syndrome: Long-term Outcomes of Sequential Steroid Therapy.

OBJECTIVE: This study aimed to investigate the long-term outcomes in children with steroid-resistant nephrotic syndrome (SRNS), who received methylprednisolone pulse therapy (MPT)-based sequential steroid therapy. In particular, we aimed to observe whether these patients had a high risk of adverse events. METHODS: We conducted a retrospective study over a 5-year period. The long-term outcomes for children with SRNS receiving sequential therapy were observed. RESULTS: Sixty-three children were diagnosed with SRNS and underwent MPT-based sequential steroid therapy. Thirty-five (55.6%) achieved complete or partial remission, 19 (30.2%) of whom were in remission even after treatment cessation at last review. The mean time to initial remission after MPT was 24.3±13.1 days. Forty-nine children (77.8%) experienced relapses, of whom 31 (49.2%) demonstrated a frequent relapsing course. Adverse effects relevant to MPT were generally mild and infrequent. Five patients (7.9%) complained of vomiting or nausea during MPT infusion; 25 (39.7%) experienced excessive weight gain and developed an obvious Cushingoid appearance; and 26 (41.3%) had poor growth associated with long-term steroid use. Twenty-eight patients (44.4%) failed to respond to MPT, of whom 21 (33.3%) achieved complete or partial remission with immunosuppressive agents. CONCLUSION: MPT-based sequential steroid therapy appears to be a safe and effective method for inducing rapid remission in childhood SRNS. Further clinical studies are needed to comprehensively evaluate this therapy.

Hashimoto's thyroiditis and papillary carcinoma in an adolescent girl: A case report.

Hashimoto's thyroiditis with thyroid cancer in childhood is not as common in the adult population. Hashimoto's thyroiditis is an autoimmune disease associated with autoantibodies, and the association between Hashimoto's thyroiditis and papillary carcinoma of the thyroid remains controversial. The present study reported a 15-year-old adolescent girl with the diagnosis of Hashimoto's thyroiditis with thyroid cancer. With the complexity of the clinical manifestations of Hashimoto's thyroiditis, it can be expressed as not only hyperthyroidism or hypothyroidism, but also normal thyroid function. The long-term treatment, and for children with thyroid cancer, early diagnosis is particularly difficult. In the present case, the diagnosis of Hashimoto's thyroiditis is primarily based on clinical manifestations, anti-thyroglobulin antibody and anti-thyroid microsomal antibody. The only diagnostic imaging ultrasound was negative. The present study discussed the possible reason and the identification of this unique case of Hashimoto's thyroiditis with thyroid cancer.

[Primary culture of human trophoblastic cells and their susceptibility to human cytomegalovirus].

OBJECTIVE: To determine the susceptibility of primary human trophoblastic cells (HTCs) to human cytomegalovirus (HCMV) infection. METHODS: 30 villus of healthy pregnant women were digested with a mixture of 0.25% trypsin and 0.2% DNase. The digestion was then filtered by a 200 mesh stainless steel sieve. HTCs were purified by Percoll gradient centrifugation and repeat purification with trypsin during cell passage. HTCs were identified by cell morphology and immunofluorescence. PCR, Western blot and immunofluorescent staining were performed to detect HCMV gene and protein 72 hours post infection of HCMV. RESULTS: The combination of 0.25% trypsin and 0.2% DNase with Percoll gradient centrifugation isolated and purified primary HTCs. More than 90 percent cells expressed CK-7 related antigen. The results of PCR, western blot and immunofluorescence showed positive expressions of CMV gB and virus proteins. CONCLUSION: HTCs is susceptible to HCMV infection, which suggests a possible route of HCMV infection at the fetal-maternal interface.

[Activation of TLR3 pathway in the pathogenesis of nephrotic syndrome induced by respiratory syncytial virus in rat model].

OBJECTIVE: To explore the activation of toll-like receptor 3 (TLR3) pathway on the process of respiratory syncytial virus (RSV) induced nephropathy. METHODS: SD rats were inoculated intranasally and intraperitoneally with 6 X 10(6) plaque forming unit(PFU) RSV and sacrificed on days 4, 14, 30 postinoculation (RSV4, RSV14 and RSV30). The normal rats without intervention were set as control. Renal tissues were obtained, and the morphological changes were studied. The expressions of TLR3, NF-kappaB and IL-13 in the rats' kidney were measured with real-time quantitative RT-PCR, and indirect IF staining. RESULTS: After the inoculation of RSV, the rats had proteinuria and the fusion of foot processes of glomerular epithelial cells, resembling human minimal changed nephrotic syndrome (MCNS). The expressions of TLR3, NF-kappaB and IL-13 in renal tissues increased obviously at Day 4 and Day 14 postinoculation, the differences were significant when compared with normal control rats (P < 0.05). The expressions of these three factors decreased at Day 30, which were not significantly different to those of normal control group (P > 0.05). CONCLUSION: TLR3 signal pathway may play an important role in early stage of RSV nephropathy.

[Primary culture of human umbilical vein endothelial cells and their susceptibility to human cytomegalovirus].

OBJECTIVE: To detect whether the primary culture of human umbilical vein endothelial cells (HUVEC) is susceptible to human cytomegalovirus (HCMV) infection so as to explain how virus from mothers traverses the placenta to fetus. METHODS: Thirty umbilical cords of healthy parturient were used in this study. The HUVEC were harvested with 0.1% collagenase for 15 minutes. The cells were identified by morphology and immunofluorescence. Then the HUVEC were infected with HCMV in vitro. The DNA of cytomegalovirus and protein of infected cells were detected 72 hours later. RESULTS: The HUVEC with adequate quality were harvested, which expressed VIII related antigen. Positive expressions of CMV gB gene and virus proteins were identified in all of the HUVEC infected with HCMV. CONCLUSION: HUVEC is susceptible to HCMV infection, which suggests a possible route of HCMV infection through the fetal-maternal interface.

[Expression of glomerular heparan sulfate domains in pediatric patients with minimal change nephrotic syndrome].

OBJECTIVES: To investigate the expression of glomerular heparin sulfate (HS) in paediatric patients with minimal change nephritic syndrome (MCNS). METHODS: The kidyney tissues were collected by biopsy from 13 paediatric patients with MCNS, while 5 normal renal biopsy samples were used as control. HS in glomeruli was analysed by indirect immunofluorescence staining using four different monoclonal antibodies, Hepss1, 3G10, JM403 and 10E4, which all recognize distinct HS species and each interacts with a specific HS domain. The concentrations of urine heparan sulfate also were measured by enzyme-linked immunosorbent assay (Elisa). RESULTS: Expression of HS fine domains was aberrant in paediatric patients compared with control subjects. Children with MCNS in replase showed a decreased glomerular expression of 10E4, JM403 and Hepss1 (P < 0.05). The level of urinary HS was significantly increased in peadiatric patients with MCNS when compared with that in control subjects (P < 0.01). CONCLUSION: These results suggest that loss of heparan sulphate in renal tissue may play a role in the pathogenesis of MCNS proteinuria.

[Effect of truncated PDGF-alpha receptor on proliferation and expression of c-sis mRNA of vascular smooth muscular cells of pulmonary artery].

OBJECTIVE: To investigate the effect of truncated PDGF-alpha receptor on proliferation expression of c-sis mRNA of pulmonary artery vascular smooth muscular cells(VSMC). METHODS: Tissue mass culture was done to get vascular smooth muscular cells of pulmonary artery. Different concentrations of truncated PDGF-alpha receptor and adenoviral recombined body (Ad5CMV-PalphaRtr, ACP) were added into the cultures. VSMC proliferation curves were obtained using MTT test. The expression of c-sis mRNA was detected by PCR. RESULTS: ACP at 5 ml/L and 10 ml/L concentrations restrained the proliferation of VSMC apparently with the peak of cell growth being attenuated or eliminated. The curve presented ascending tendency only after 7 days. Affected by 5 ml/L ACP, PDGF-BB exerted no significant accelerating effect on the proliferation of VSMC. The expression of c-sis mRNA was up-regulated under the effect of ACP. Affected by 5 ml/L ACP and PDGF-BB, the expression of c-sis mRNA was down-regulated. CONCLUSION: ACP at 5 ml/L and 10 ml/L concentration powerful inhibitor of cellular proliferation for pulmonary artery VSMC. It can increase c-sis mRNA expression significantly and the increase seems to be ACP dosage dependent.

[The change of angiotensin II receptors in children with left-to-right shunt].

OBJECTIVE: To explore the change of AT1 and AT2 receptors in children with left-to-right shunt. METHODS: Lung tissue was obtained from 20 children with left-to-right shunt by biopsy during operation, and lung tissue from 5 children by autopsy was used as control. The above specimens were stained by immunohistochemistry techniques for detecting AT1 and AT2 receptors. Histomorphometric study was done to measure the IOD value of pulmonary small vascular wall via the staining of AT1 and AT2 receptors, and the diameter of these vessels was 15-100 microm. RESULTS: The IOD values of pulmonary small vascular walls obtained from via the staining of AT1 and AT2 receptors in children with left-to-right shunt were higher than the IOD values in control group (P=0.000). CONCLUSION: The expression of AT1 and AT2 receptors in pulmonary small vascular walls in children with left-to-right shunt was higher than that of the control. It may provide a sound rationale for the pharmacological management of children with left-to-right shunt by use of ACEI or AT1 receptor antagonist.

[Expression and variation of angiotensin II type 1 receptor on vascular smooth muscle cells of systemic and pulmonary circulations in experimental ventricular septal defect in pigs].

OBJECTIVE: To investigate the expression of angiotensin II type 1 receptor (AT1R) on the vascular smooth muscle cells (VSMC) of both systemic and pulmonary circulations and their variation caused by intracardiac left to right shunting using an animal model of the ventricular septal defect (VSD). METHODS: Nineteen young pigs were divided into 3 groups: operation (experimental VSD), sham-operation, and normal control. One month after operation, the pigs were catheterized and then put to death. Smooth muscle cells were taken from aorta, pulmonary artery, pulmonary arteriole, mesenteric arteriole and VSMC were isolated. Radioligand binding assay for AT1R was done to measure the Bmax and KD. RESULTS: There were no significant differences in Bmax and KD of AT1R between the sham-operation group and the control. Bmax of the aorta (112.11+/-35.77) fmol/10(6), main pulmonary artery (52.37+/-31.09) fmol/10(6) and mesenteric arteriole (106.98+/-100.48) fmol/10(6) in the operation group were remarkably elevated in comparison with the control (P<0.05). In the operation group, Bmax of the aorta VSMC was higher than that of main pulmonary artery VSMC (P<0.05), and Bmax of the mesenteric arteriole VSMC was higher than that of the small pulmonary arteriole VSMC (P<0.05). CONCLUSION: The findings in AT1R expression in experimental VSD were supportive to the previously proposed hypothesis that ATR expression might be stronger in systemic VSMC than in pulmonary VSMC in young animal, especially in the presence of intracardiac left to right shunt, which may provide a sound rationale for the pharmacological management of VSD infants with angiotensin converting enzyme inhibitor or AT1R antagonist.

[Effect of truncated platelet-derived growth factor-alpha receptor on apoptosis and expression of c-sis mRNA of pulmonary artery smooth muscle cells].

OBJECTIVE: Platelet-derived growth factor (PDGF) plays an important role during the pathophysiological changes in vascular remodeling. The study aimed to investigate the effect of truncated PDGF-alpha receptor on apoptosis and expression of c-sis mRNA of pulmonary artery smooth muscle cells (VSMCs). METHODS: Tissue mass culture was done to get vascular smooth muscle cells of pulmonary artery in newborn pigs. Two methods were used to interfere VSMCs: adding adenoviral recombined body (Ad5CMV-PalphaRtr, ACP) with three different concentrations of truncated PDGF-alpha receptor into the cultures, or adding three concentrations of PDGF-BB after the treatment with mid-concentration of ACP. VSMC apoptosis, cellular cycle and expression of c-sis were observed using flow-cytometry, and the expression of c-sis mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: ACP with mid- to- high concentrations could restrain the proliferation of VSMCs apparently with the increase of G(0)/G(1) cells. The apoptotic rate presented an ascending tendency. The differences among the groups were of statistically significant. Affected by mid- concentration of ACP, PDGF-BB did not exhibit a significantly accelerating effect on the changes of cellular cycle and VSMC apoptosis. The expression of c-sis mRNA was up-regulated under the effect of ACP. Affected by mid-concentration of ACP and PDGF-BB, c-sis mRNA expressed was down-regulated. CONCLUSION: Mid- to- high concentration of ACP is a powerful inhibitor of cellular proliferation for pulmonary artery VSMCs. It can significantly increase cells in number in G(0)/G(1) phase, apoptosis and c-sis mRNA expression.