Biosynthesis of steroidal intermediates using Mycobacteria: a review / 生物工程学报

Steroids are a class of medicines with important physiological and pharmacological effects. In pharmaceutical industry, steroidal intermediates are mainly prepared through Mycobacteria transformation, and then modified chemically or enzymatically into advanced steroidal compounds. Compared with the "diosgenin-dienolone" route, Mycobacteria transformation has the advantages of abundant raw materials, cost-effective, short reaction route, high yield and environmental friendliness. Based on genomics and metabolomics, the key enzymes in the phytosterol degradation pathway of Mycobacteria and their catalytic mechanisms are further revealed, which makes it possible for Mycobacteria to be used as chassis cells. This review summarizes the progress in the discovery of steroid-converting enzymes from different species, the modification of Mycobacteria genes and the overexpression of heterologous genes, and the optimization and modification of Mycobacteria as chassis cells.

Case report of lung transplantation for cystic fibrosis in a child / 中华实用儿科临床杂志

Objective:A case of cystic fibrosis admitted in the Respiratory Department of Beijing Children′s Hospital, Capital Medical University in June 2018 and underwent lung transplantation later was analyzed retrospectively.A 10-year-old girl had intermittent productive cough for more than 4 years with clubbed-finger.The lung high resolution CT (HRCT) showed bronchiectasis and mucus impaction, and the nasal sinus HRCT showed sinusitis.She had cystic fibrosis transmembrane conductance regulator ( CFTR) gene complex heterozygous mutation and positive sweat test.The immunoglobulin E (IgE) level and eosinophil count increased, and aspergillus fumigatus-specific IgE was positive.She was diagnosed as cystic fibrosis, allergic bronchopulmonary aspergillosis and sinusitis.Anti-infection, glucocorticoid and symptomatic treatment were given.During the following 21 months, the child had repeated respiratory difficulties and respiratory failure.Her lung function declined.Bilateral lung transplantation was performed in March 2020.She had a good recovery at more than 1 year and 3 months postoperatively.

Advance in Niemann-Pick disease type C / 国际儿科学杂志

Niemann-Pick disease type C is a lipid storage disorder associated with impaired intracellular cholesterol trafficking, caused by mutations of either NPC1 or NPC2 genes.According to the age at onset of symptoms, it is divided into 5 categories, including neonatal, early-infantile, late-infantile, juvenile and adult type.There are differences in clinical manifestations and prognoses among each category.The characteristic clinical manifestations are hepatosplenomegaly, lung infiltration, vertical supranuclear gaze palsy and gelastic cataplexy.The definite diagnosis requires demonstration of unesterified cholesterol accumulated in fibroblasts cultured from skin biopsies with filipin staining and(or) of pathogenic mutation of NPC1/NPC2 genes.There is no effective treatment for this disease yet, therefore the overall prognosis is still poor.Miglustat can delay onset of the neurological symptoms, and prolong survival of partial patients.

Clinical and genetic characteristics and differential diagnosis in children with autosomal dominant hyper-IgE syndrome / 中华实用儿科临床杂志

Objective:To summarize the clinical features of children with autosomal dominant hyper-IgE syndrome (AD-HIES) and the differential diagnosis of hyper-IgE syndrome and allergic diseases as well.Methods:All clinical data, including general information, clinical features, and genetic changes, from 7 children with AD-HIES who were diagnosed in Beijing Children′s Hospital Affiliated to Capital Medical University from April 2016 to June 2020 were analyzed retrospectively.The diagnostic criteria are based on the National Institutes of Health′s (NIH)′s hyper-IgE syndrome score and combined with the results of gene detection, shown as follows: (1) NIH score over 40, with signal transducer and activator of transcription 3 gene ( STAT3) pathogenic mutation; (2) NIH score between 20 and 40, with reported STAT3 pathogenic mutation; (3) NIH score less than 20 points was excluded. Results:There were 3 males and 4 females.The onset age of 7 cases was within 2 months after birth, and the mean age at diagnosis was 3 years old.All seven cases had recurrent skin or lung infections, with 4 cases having skin and lung infections, 1 case of skin abscesses at the BCG vaccination site, and 2 cases without skin infection suffering from recurrent pneumonia.The mean onset age of skin abscess in 5 cases was 1.5 years, and pus culture of 3 cases were Staphylococcus aureus.Four cases developed bullae and 6 cases had lung infections.Four cases had otitis media, and oral thrush was seen in 4 cases.One case of skin and lung infection developed liver abscess and sepsis.Seven cases had eczema, which was disco-vered in the neonatal period for 6 cases.Four cases had the symptoms of eczema for the first visit.Two cases had food allergy, and 1 case had recurrent wheezing within 1 year old.The serum IgE level and blood eosinophil count in 7 children were elevated.All children had heterozygous pathogenic mutations in STAT3.Six patients had de novo mutations.There were 6 different mutation sites.The 4 mutation sites were reported: c.1145G>A, c.1144C>T, and c. 1699A>G were missense mutations, and c. 1139+ 5G>A was splicing mutation.Two mutation sites had not been reported: c.1031A>C was missense mutation, and c. 2050G>T was nonsense mutation.The pathogenic grade of them were likely pathogenic, and the NIH score of 2 cases were above 40 score, which was consistent with the clinical diagnosis of hyper-IgE syndrome. Conclusions:Eczema is a common and early clinical manifestation of hyper-IgE syndrome, along with elevated IgE levels and eosinophil counts that need to be differentiated from allergic diseases.On the contrary, it often had recurrent skin abscesses or pneumonia, which was prone to bullae.The clinical manifestations of young children were atypical, and genetic testing was helpful for early diagnosis.

Periosteum-covered iliac crest autografts for treatment of severe osteochondral lesions of talus / 中华创伤杂志

Objective:To explore the clinical efficacy of periosteum-covered iliac crest autografts for treatment of severe osteochondral lesions of talus （OCLTs）.Methods:A retrospective case series study was used to analyze the clinical data of 26 patients with severe OCLTs treated at Zhejiang Armed Police Corps Hospital from January 2013 to October 2019. There were 21 males and 5 females，aged 17-49 years ［（36.3 ± 10.9）years］. All patients were treated using periosteum-covered iliac crest autografts. The visual analogue scale （VAS），American Orthopedic Foot and Ankle Society （AOFAS） ankle-hindfoot score and ankle joint range of motion （ROM） were assessed before operation，6 months after operation and at the last follow-up （≥ 12 months）. The area of talus injury with MRI at the same level was recorded before operation and at the last follow-up. The healing of talus and joint surface was detected with CT at the last follow-up. The healing of the incision and osteotomy site and complications were observed.Results:All patients were followed for 12 to 22 months［（15.1 ± 3.2）months］. The VAS was （2.4 ± 0.9）points and （1.7 ± 0.6）points at postoperative 6 months and at the last follow-up，significantly lower than the preoperative （5.4 ± 1.2）points （ P < 0.01）. Meanwhile，the VAS at the last follow-up was significantly lower than that at postoperative 6 months （ P < 0.01）. The AOFAS ankle-hindfoot score was （71.7 ± 7.8）points and （87.8 ± 6.2） points at postoperative 6 months and at the last follow-up，significantly lower than the preoperative （66.5 ± 7.5） points （ P < 0.01）. Meanwhile，the AOFAS ankle-hindfoot at the last follow-up was significantly lower than that at postoperative 6 months （ P < 0.01）. The ankle ROM was （58.4 ± 5.5）° and （70.0 ± 4.9）° at postoperative 6 months and at the last follow-up，significantly improved when compared to the preoperative （42.3 ± 8.1）° （ P < 0.01）. Meanwhile，the ankle ROM at the last follow-up was significantly improved when compared to that at postoperative 6 months （ P < 0.01）. The area of talus injury with MRI at the same level was 0.67（0.55，0.89）cm 2 at the last follow-up，significantly improved when compared to preoperative 2.64（1.98，3.68）cm 2 （ P < 0.01）. The transplantation had neither obvious defects nor joint surface steps based on CT findings. All surgical incisions were healed by first intention. There were no complications such as incision infection，skin necrosis，nonunion of osteotomy，malunion or severe ankle joint disorder except that 8 patients had residual local subchondral bone?marrow?edema-like?signal?and 2 patients showed delayed healing of medial malleolus osteotomy. Conclusion:For patients with severe OCLTs，periosteum-covered iliac crest autografts can effectively relieve ankle pain，improve ankle function，and reduce the area of injury.

Primary immunodeficiency diseases susceptible to mycobacteria in children / 中华实用儿科临床杂志

Primary immunodeficiency diseases (PIDs) refer to immune function decrease and deficiency or immune regulation function imbalance resulted from immune cell or immunomolecular defects caused by single gene mutation.Children with PIDs are prone to develop infectious diseases.Because the immune mechanisms of hosts infected by different pathogens infecting vary, different types of PIDs are relatively susceptible to different pathogens.A number of studies have indicated that some PIDs children are more likely to develop Bacilles Calmétte-Guerin infection, severe tuberculosis and non- Tuberculous mycobacteria severe infection.In this review, several major primary immunodeficiency diseases closely related to the susceptibility of mycobacteria were summarized, including chronic granulomatous disease, severe combined immunodeficiency disease, mendelian susceptibility to mycobacterial disease, and high IgM syndrome, in order to provide a theoretical basis for early identification of such diseases.

A single-center study of pneumococcal empyema in children / 中华实用儿科临床杂志

Objective:To understand the clinical characteristics of empyema caused by Streptococcus pneumoniae in children. Methods:The clinical manifestations, imaging characteristics, treatment and prognosis of 49 children with pneumococcal empyema admitted to Beijing Children′s Hospital, Capital Medical University from March 2007 to February 2018 were retrospectively analyzed.Results:Among the 49 children, 26 were male and 23 were female, with a median age of 2.63 years old.All the cases had cough and fever, 46 cases of them had high fever, 2 cases had moderate fever and 1 case had ultrahyperpyrexia.The course of disease before admission was 2-90 days, with a median of 10 days.All cases had toxic symptoms and signs of pleural effusion; 26 patients had dyspnea; 23 patients had moist rales, while 8 cases of whom had wheezing sounds.Eleven cases had extrapulmonary complications, including purulent meningitis in 6 cases, purulent pericarditis 1 case, 2 cases of hemophagocytic syndrome and hemolytic uremic syndrome respectively.The medians of leukocytes in the whole blood, white blood cells in pleural effusion and multinuclear cell percentage were 26.97×10 9/L, 32×10 9/L and 0.85, respectively.The average C-reactive protein (CRP) was 177.79 mg/L.The drug sensitivity test of Streptococcus pneumoniae showed that 68.89% strains were insensitive to Penicillin and all strains were resistant against Erythromycin.Chest image showed bilateral consolidation in 32 cases and pneumothorax in 31 cases.Besides antimicrobial therapy, 34 patients were treated with chest drainage, and 7 cases underwent surgery.Ten of the 49 cases were treated with nasal continuous positive airway pressure, tracheal intubated was used in 6 cases, and 1 case was given cardiopulmonary resuscitation.Forty-five cases discharged with improvement, 3 cases were not cured and 1 case died. Conclusions:Pneumococcal empyema is more common in children under 5 years old.Children with pneumococcal empyema usually have poor mental status, high fever, cough accompanied by dyspnea, high peripheral white blood cells and CRP.Their radiographic findings are usually serious, and the insensitive rate to Penicillin is high.The main treatment is anti-infection therapy and closed thoracic drainage.The prognosis of most patients is good, but there are still dead cases.

Clinical characteristics of community-acquired influenza virus pneumonia in hospitalized children / 中华实用儿科临床杂志

Objective To analyze the clinical characteristics of community-acquired influenza virus pneumonia in hospitalized children and improve the clinicians' understanding level of this disease.Methods Data of 70 cases with community-acquired influenza virus pneumonia admitted to the Respiratory Department and Infectious Disease,Beijing Children's Hospital,Capital Medical University,from November 2009 to April 2018 were collected and the clinical characteristics were analyzed.Results Of the 70 cases,61 cases(89.7％) were discharged after improvement.The median age was 3.5 years old,and 50 cases(71.4％) were 0 to 5 years old.There were 29 cases with severe influenza pneumonia,41 cases with mild influenza pneumonia,3 cases died,and 19 cases (27.1％) had underlying diseases.Sixty-four cases (91.4％) were hospitalized in winter and spring.The first symptoms were mainly fever in 64 cases (91.4％) and cough in 65 cases (92.9％),and temperatures were mostly from 39.1 ℃ to 41.0 ℃.Lung auscultation was dominated by moist rales (30 cases,58.8％) and wheezing (8 cases,15.7％).There were many complications of influenza virus pneumonia,including 19 cases with myocardial injury,11 cases with liver function injury,4 cases with toxic encephalopathy,3 cases with electrolyte disturbance,2 cases with multiple organ failure,2 cases with hemophagocytic syndrome,and 1 case with septic shock.Chest radiographic results reveal bilateral inflammation in 40 children (57.1％),prodominatly in lower lobe lesions (39 cases).The common changes were patchy shadow,interstitial parenchymal lesion,ground glass shadow,and pleural effusion.Forty-seven children (67.1％) were infected by influenza A,and 23 children(32.9％) were co-infected.The percentage of severe cases with underlying diseases (68.4％) was significantly higher than that in children without chronic diseases (31.4％),the difference was statistically significant (x2 =7.830,P =0.005).The increase rate of C reaction protein (CRP) in severe cases (54.3％) was significantly higher than that in mild cases (28.6％),the difference was statistically significant (x2 =4.769,P =0.029).Conclusions Community-acquired influenza virus pneumonia in children mainly occurs in winter and spring.It is more common seen in children under 5 years of age.The main clinical manifestations of community-acquired influenza virus pneumonia are high fever and cough,extrapulmonary complications are more common.Most children have moist rales and showed bilateral inflammation and lower lobe lesions in chest radiography.Children with underlying diseases are more likely to develop severe influenza virus pneumonia.Elevated CRP is associated with severe influenza virus pneumonia.Most patients have a good prognosis,but there are still cases of death.

Recurrent petechia and epistaxis in a patient diagnosed as diabetes mellitus for 2 years / 中华实用儿科临床杂志

To analyze respectively a case,presented with recurrent petechial and epistaxis after a 2 years history of diabetes mellitus (DM),who was hospitalized in Beijing Children's Hospital Affiliated to Capital Medical University.The clinical manifestation,examination,diagnosis and treatment were recorded.The patient was diagnosed with immune thrombocytopenia (ITP) and DM at the first admission.The initial therapy with gamma globulin didn't show ideal effect.The pediatric specialists from the department of ENT,Hematology/Oncology,Endocrinology,Pharmacy and Immunodeficiency Clinic were invited to discuss the case.The final diagnosis of autoimmune polyglandular syndrome (APS) was made and supplementary steroid treatment was started.But the response of the steroid therapy was poor.Once again with the multidisciplinary consultation,the patient received several schemes of Rituximab under the informed consent.This treatment reached a stable condition for almost 7 years.APS should be considered when DM patient showed the manifestation of other immune organ damages.Rituximab immunosuppressive therapy should be tried when the response to first-line treatment was poor.

A case-control study on the relationship between serum FGF-21 and coronary heart disease / 实用医学杂志

Objective To investigate the association of serum fibroblast growth factor 21(FGF-21)with coronary heart disease(CHD)and biochemical indexes. Methods Serum FGF-21 expression was detected by ELISA.310 healthy control subjects and 310 CHD patients were recruited.Basic information was obtained by clini-cal questionnaires. Venous blood was tested for serum lipid,fasting glucose,and C-reactive protein(CRP). All the data were analyzed with the SPSS 19.0. Results There were no significant differences in gender and age be-tween CHD and controls(P > 0.05). FGF-21 level was significantly higher in CHD patients than in the controls (P<0.05).Univariate analysis showed serum FGF-21 was a risk factor of CHD.The group of higher score for FGF-21 had a 5.20-fold risk of CHD than the group of lower score without adjustment for confounding factors(95% CI:4.23~8.42).After control of the confounding factors,the group of higher score had a 7.21-fold risk of CHD(95% CI:3.78~13.67)than the group of lower score,it remains the significant difference(P<0.05).Conclusion Ele-vation of serum FGF-21 was closely associated with initiation and development of CHD.

Congenital defects of phagocyte number or function / 中华实用儿科临床杂志

Congenital defects of phagocyte number,function or both was categorized to the fifth classification from the international union of immunological societies expert committee for primary immunodeficiency 2015.Severe congenital neutropenia was the most fatal phagocyte number defect.Phagocyte functions included motility,chemotaxis,adhesion,phagocytosis and killing.Leukocyte adhesion deficiency and chronic granulomatous disease were the most common diseases.This article will describe pathogenesis,molecular,clinical,laboratory features and treatment and prognosis,to supporting clues for paediatrician's clinical operations.

22q11.2 deletion syndrome / 中华实用儿科临床杂志

22q11.2 deletion syndrome is the commonest chromosome deletion syndrome.Most patients with DiGeorge anomaly have monosomic deletions of chromosome 22q11.2.Abnormal pharyngeal arch development results in defects in the development of the parathyroid glands,thymus and conotruncal region of the heart.Defective thymus development is associated with impaired immune function." Complete" DiGeorge syndrome accounts for ＜ 0.5％ of patients with total absence of the thymus and a severe T cell immunodeficiency.Most patients with partial defects have variable T cell deficiency.There is a wide phenotypic spectrum including speech delay,neuropsychiatric disorders and otolaryngological disorders.These patients are at increased risk of a variety of autoimmune diseases.Severe cases need thymus transplantation.

Gain-of-function signal transducer and activator of transcription 1 deficiency / 中华实用儿科临床杂志

Chronic mucocutaneous candidiasis (CMC) is characterized by persistent or recurrent disease of the nails,skin,oral,or genital mucosae caused by candida albicans.CMC usually can occur in patients with T cell deficiencies,autosomal dominant hyper-immunoglobulin E(IgE) syndrome,interleukin(IL)-12p40 and IL-12 receptor β1 (IL-12Rβ1) deficiency,caspase recruiment domain 9 deficiency and autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy.CMC pathogenesis apparently involves the impairment of IL-17A,IL-17F and IL-22 immunity.Autosomal recessive IL-17RA deficiency and dominant-negative IL-17F deficiency are etiologies of pure isolated CMC (CMCD).Nearly half of patients with CMC had gain-of-function signal transducer and activator of transcription 1 mutations.These patients also had bacterial and virus infections,autoimmunity and inflammatory diseases,which show broad clinical heterogenity.

Mendelian susceptibility to mycobacterial disease / 中华实用儿科临床杂志

Mendelian susceptibility to mycobacterial diseases (MSMD)is a rare congenital disorder characterized by susceptibility to poorly virulent mycobacteria,such as Bacille Calmette-Guerin vaccine or non-tuberculous environmental mycobacteria.The interferon-γ'(IFN-γ)/interleukin-12 (IL-12) pathway is central to controlling mycobacterial infections,in which several genes had been identified.IFN-γ secretion is impaired in patients with IL-12p40 and IL-12 receptor β1 deficiency,where the response to IFN-γ is impaired in patients with IFN-γ receptor 1,IFN-γ receptor 2,and signal transducer and activator of transcription 1 deficiencies.Furthermore,germline mutations in the cytochrome b (-245) beta subunit,interferon regulatory factor 8,ubiquitin-like modifier,RORC and TYK2 have been identified as the genes which are responsible for MSMD.These patients do not generally have associated infections,apart from salmonellosis.Now,the pathogenesis,molecular,clinical,laboratory features,treatment and prognosis were described,in order to support the clues for pediatrician's clinical practice.

Activated phosphoinositide 3-kinase δ syndrome / 中华实用儿科临床杂志

Activated phosphoinositide 3-kinase δ (PIK3CD) syndrome is a combined immunodeficiency,caused by PIK3CD gain-of-function mutations,which encodes the catalytic subunit of PIK3CD.These patients presented with early onset sinopulmonary infections,lymphoproliferation,herpesvirus infections,autoinflammatory disease,lymphoma and mental retardation.Hyper immunoglobulin (Ig)M,IgG deficiency,CD4 lymphopenia were common immunologic features.Variable expression can lead to death and asymptom.Penetrance can be incomplete.Hematopoietie stem cell transplantation should be taken into account for severe cases.

Combined immunodeficiency / 中华实用儿科临床杂志

Combined immunodeficiency (CID) is categorized to the first classification from the international union of immunological societies expert committee for primary immunodeficiency.Severe combined immunodeficiency (SCID) is the most fatal disorder for paediatric clinical operation.Without hematopoietic stem cell transplantation,almost all infants would die before 1 year old,few could survive beyond 2 years old.Hypomorphic mutations in SCID genes can lead to atypical phenotypes.The two special SCID should be focused,Omenn syndrome and graft-versus-host disease,which are caused by expension of autologous and maternal activated and memory T lymphocytes,respectively.Patients with radiosensitive-CID usually present later on life,for whom treatment should be monitored carefully.CID caused by T cells with normal development and inborn error was hotspot research field for example zeta chain-associated protein 70 kDa deficiency.More attention should be paid to CID associated with syndromes for example dedicator of cytokinesis 8 deficiency.Now,the pathogenesis,molecular,clinical,laboratory features and treatment and prognosis are described,in order to support clues for paediatrician's clinical practice.

Diagnosis and treatment of atypical severe combined immunodeficiency disease in 7 children / 临床儿科杂志

Objective To explore the diagnosis and treatment of atypical severe combined immunodeficiency disease (SCID). Methods The clinical data of atypical SCID in 7 children with IL2RG,JAK3,and RAG1 mutations were reviewed and analyzed from September 2012 to June 2017. Results In 7 cases (6 males and 1 female), there were 5 infants, 1 toddler and 1 school-age child. Cases 2, 4, and 6 were classic SCID clinical phenotypes. Cases 1, 3, 5, 7 were atypical SCID clinical phenotypes. Case 6 were diagnosed with Omenn syndrome. Cases 2, 5 were classic SCID immune phenotypes, cases 1, 3, 4, 6, 7 were atypical SCID immune phenotypes, and case 1 had maternal chimera. The next generation sequencing indicated that case 1 had a compound heterozygous JAK3 mutation with c.3097-1G>A/c.946-950GCGGA>ACinsGGT.Cases 2,3,and 4 had IL2RG mutations,with c.865C>T/p.R289X,c.664C>T/R222C,52delG,respectively.Case 5 had JAK3 mutations with c.2150A>G/p.E717G and c.1915-2A>G.Sanger sequencing indicated that case 6 had a RAG1 mutation of complex heterozygosity with c.994C>T/p.R332X and c.1439G>A/p.S480N. Case 7 had homozygous RAG1 mutation with c.2095C>T/p.R699W.Conclusion Under certain conditions,gene mutation can lead to atypical clinical and/or immune phenotypic SCID.

Chronic granulomatous disease and Mcleod syndrome caused by continuous X chromosome deletion：a report of two cases and literature review / 临床儿科杂志

Objective To explore the clinical features of chronic granulomatous diseases and Mcleod syndrome caused by continuous X chromosome deletion. Methods The clinical data of two children diagnosed as chronic granulomatous disease and Mcleod syndrome by gene detection were retrospectively analyzed. Results Two males, 4 year 1 month and 1 year 9 month old, were both hospitalized due to persistent pulmonary infections. Both of them had a history of repeated severe infections and BCG vaccine associated lymphadenitis, and were diagnosed as X-linked chronic granulomatous disease for respiratory burst defects and deletion of all CYBB exons. Both of them had retarded motor development, and were diagnosed as DMD for detection of DMD gene exons and muscle speciifc promoter region and exon 1-2 deletion by MLPA. One case was found with obvious echinocytes, the other case showed whole exons deletion of XK gene. Both of them were diagnosed as Mcleod syndrome. Conclusion Continuous X chromosome deletion could lead to combination of Mcleod syndrome, DMD, and X-CGD, which may complicate the condition. Due to the lack of Kx antigen, repeated common blood transfusion can produce relative antibody, which lead to severe hemolytic crisis.

Influence of transcatheter closure combined drug intervention on BNP and vascular vascular endotheli-um levels in children with CHD complicated pulmonary artery hypertension / 心血管康复医学杂志

Objective:To study the influence of transcatheter closure combined drug intervention on levels of brain na-triuretic peptide (BNP)and vascular endothelium (ET)in children with congenital heart disease (CHD)complicated pulmonary artery hypertension (PAH).Methods:A total of 70 CHD + PAH children from Mar 2011 to May 2013 in our hospital were enrolled.According to random number table,they were equally divided into transcatheter clo-sure combined drug intervention group (combined treatment group)and transcatheter closure control group (only re-ceived transcatheter closure therapy).Cytokine levels and heart function indexes were observed and compared be-tween two groups.Results:Compared with transcatheter closure control group after treatment,there were signifi-cant reductions in levels of BNP [(352.7±56.2)ng/L vs.(194.5±25.2)ng/L]and vascular ET [(68.4±8.4)ng/L vs.(37.5±5.2)ng/L,P <0.01],and significant rise in left ventricular early diastolic peak filling velocity [E, (63.7±6.9)cm/s vs.(71.7±7.4)cm/s],left ventricular early/late diastolic peak filling velocity [E/A,(1.10± 0.13)vs.(1.34±0.16)],left ventricular ejection fraction [LVEF,(62.3±7.6)% vs.(66.4±7.1)%]and left ventricular end-diastolic diameter [LVEDd,(48.9±5.7)mm vs.(54.1±5.8)mm]in combined treatment group, P <0.05 or <0.01.Conclusion:Transcatheter closure combined drug intervention can reduce BNP and vascular ET concentrations,improve heart function,which possesses positive clinical application value.

Sodium-calcium exchanger promoter promotes differentiation of mouse induced pluripotent stem cells into cardiomyocytesin vitro / 中国组织工程研究

BACKGROUND:Inducing pluripotent stem cels has been considered as a promising treatment for ischemic heart disease. However, an ideal inducing method has not been found yet. OBJECTIVE:To investigate the role of sodium-calcium exchanger (NCX1) promoter in the differentiation of mouse induced pluriptent stem cels (miPS) into cardiomyocytes. METHODS: The pLVX-IRES-ZsGreen1 vectors which contain NCX1 promoter constructed by recombinant DNA technology were co-transfected to 293FT cels with ViraPowerTM Lentiviral Packaging Mix. The recombinant lentiviruses infected with miPS were selected and purified by puromycin. miPS were recovered and passaged to form embryoid bodies. The embryoid bodies were induced by differentiation medium containing various concentrations of the virus titer. The number of beating embryoid bodies were calculated. The expression profiles of the myocardial intra-markers were tested to determine the differentiation efficiency of iPSC by RT-PCR and immunofluorescence analysis. RESULTS AND CONCLUSION:pLVX-IRES-ZsGreen1 vectors which contain NCX1 promoter were constructed and confirmed by PCR. Virus could be packaged, purified and concentrated successfuly. The recombinant lentivirus to transduce miPS was sorted by flow cytometry. In contrast to NCX1-/GFP- cels, NCX1+/GFP+ cels were differentiated and developed prominent beating areas with sustained contractile activity for additional 4 days, and demonstrated positive expression of gap communication marker CX43 and cardiac troponin. The expressions of GATA4, MEF2c and Nkx2.5 in the NCX1+ cels were 4.2, 7.5, and 2.5 times those in NCX1- cels. Results showed the NCX1 promoter can promote the cardiac differentiation of miPS .