Generation of an induced pluripotent stem cell line from a patient with surfactant metabolism dysfunction carrying ABCA3 mutations / 中华实用儿科临床杂志

Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.

Correlation of immunoglobulin variable heavy chain gene mutation status with prognosis in patients with mantle cell lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the relationship between immunoglobulin variable heavy chain (IgVH) gene mutation status and clinical features, pathologic findings and biologic behavior of mantle cell lymphoma (MCL).</p><p><b>METHODS</b>IgVH gene was amplified in 60 cases of MCL with FR1-JH and FR2-JH primers in BIOMED-2. The sequence was determined by cloning. The IgVH somatic mutational status was analyzed using NCBI's Ig-Blast tool. The relationship between IgVH gene mutation status and clinicopathologic features was also analyzed.</p><p><b>RESULTS</b>Forty percent (24 cases, 28 functional Ig genes) of the MCL cases displayed somatically mutated VH genes (defined as > 2% mutated), whereas 60.0% (36 cases, 40 functional Ig genes) showed unmutated VH genes. The most widely used genes were VH3-21 (27.9%) and VH4-34 (19.1%). The former were mainly used by unmutated cases, while the later mainly by mutated cases.Intraclonal heterogeneity was noted in 19 cases. There was no correlation of VH mutation status and specific VH gene with survival (P > 0.05).</p><p><b>CONCLUSIONS</b>MCL comprises at least two subsets that do not correlate with morphology: one with unmutated VH genes and one with mutated VH genes. The biased use of VH3-21 and VH4-34 is noted. The nonrandom usage of IgVH segments suggests specific antigens may play a role in the pathogenesis and progression of MCL subsets. There is no correlation of IgVH mutation status and specific VH gene with survival.</p>

Effect of Speech Therapy on Articulation Disorder of Children with Cerebral Palsy / 中国康复理论与实践

@#Objective To explore the effect of speech therapy on articulation disorder of indifferent kinds of children with cerebral palsy(CP).Methods 49 CP children with articulation disorder were divided into the common group(n=21)and family rehabilitation group(n=28).All children in the two groups were treated with systemic speech therapy,but those in the family rehabilitation group were added with family rehabilitation.The changes of articulation disorder of children in two groups before and after treatment were observed.Results After treatment,all children got improvement,but the effect of children with spastic type was superior to those with other CP types.The efficiency rate of the family rehabilitation group was 39.3%,that of the common group was 14.3%,there was a significant difference between two groups(P<0.05).Conclusion The children with different CP type have different therapeutic effects for articulation disorder,the family rehabilitation can improve the therapeutic effect.

Preventive effects of aminophylline on the pulmonary hypertension rebound reaction to exposure to NO in the hypoxic pigs / 中国病理生理杂志

AIM: To evaluate the prophylactic effect of aminophylline on the pulmonary hypertension rebound reaction to exposure to NO in the hypoxic pigs. METHODS: The 10 pigs undergone Swan-Ganz catheter, the mPAP was measured with a Physio-recording instrument and PaO 2 was measured with a blood gas analyzer, when breathing NO for 30 minutes and suddenly stopping breathing NO, administing aminophylline 0.25 g, followed by 30 minutes with room air. The respiratory rate and heart rate were also monitorried with a Hewlett-Packard portable monitor. RESULTS: The mPAP of the acute hypoxic pig was induced significantly after breathing 10 -5 NO. When suddenly stopping breathing NO, the induced mPAP became more and more high, the level of the mPAP in 5 minutes was similar to the values before absorbing NO, the mPAP in 10 minutes was higher than values before absorbing NO, while the level of PaO 2 was lower than the values before absorbing NO; but suddenly stopping breathing NO, administing aminophylline, although the induced mPAP became high, the speed was slower than the controls, the level of the mPAP in 30 minutes still was lower than the values before absorbing NO. CONCLUSION: Aminophylline has preventive effects on the pulmonary hypertension rebound reaction to exposure to NO in the hypoxic pigs.