Influencing factors of survival of patients with airway stenosis requiring clinical interventions after lung transplantation / 器官移植

Objective To analyze the influencing factors of survival of patients with airway stenosis requiring clinical interventions after lung transplantation. Methods Clinical data of 66 patients with airway stenosis requiring clinical interventions after lung transplantation were retrospectively analyzed. Univariate and multivariate Cox’s regression models were adopted to analyze the influencing factors of survival of all patients with airway stenosis and those with early airway stenosis. Kaplan-Meier method was used to calculate the overall survival and delineate the survival curve. Results For 66 patients with airway stenosis, the median airway stenosis-free time was 72 (52,102) d, 27% (18/66) for central airway stenosis and 73% (48/66) for distal airway stenosis. Postoperative mechanical ventilation time [hazard ratio (HR) 1.037, 95% confidence interval (CI) 1.005-1.070, P=0.024] and type of surgery (HR 0.400, 95%CI 0.177-0.903, P=0.027) were correlated with the survival of patients with airway stenosis after lung transplantation. The longer the postoperative mechanical ventilation time, the higher the risk of mortality of the recipients. The overall survival of airway stenosis recipients undergoing bilateral lung transplantation was better than that of their counterparts after single lung transplantation. Subgroup analysis showed that grade 3 primary graft dysfunction (PGD) (HR 4.577, 95%CI 1.439-14.555, P=0.010) and immunosuppressive drugs (HR 0.079, 95%CI 0.022-0.287, P<0.001) were associated with the survival of patients with early airway stenosis after lung transplantation. The overall survival of patients with early airway stenosis after lung transplantation without grade 3 PGD was better compared with that of those with grade 3 PGD. The overall survival of patients with early airway stenosis after lung transplantation treated with tacrolimus was superior to that of their counterparts treated with cyclosporine. Conclusions Long postoperative mechanical ventilation time, single lung transplantation, grade 3 PGD and use of cyclosporine may affect the survival of patients with airway stenosis after lung transplantation.

Development status and new challenges of heart transplantation / 器官移植

Heart transplantation is the primary therapeutic option for patients with end-stage heart failure. The shortage of donors has been the main limiting factor for the increasing quantity of heart transplantation. With persistent updating and introduction of novel technologies, the donor pool has been increasingly expanded, such as using the heart from older donors, donors infected with hepatitis C virus, donors dying from drug overdose or donation after cardiac death (DCD) donors, etc. Meantime, the proportion of recipients with advanced age, multiple organ dysfunction, mechanical circulatory support and human leukocyte antigen antibody sensitization has been significantly increased in recent years. The shortage of donors, complication of recipients' conditions, individualized management of immunosuppressive therapy and prevention and treatment of long-term cardiac allograft vasculopathy are all challenges in the field of heart transplantation. In this article, novel progresses on donor pool expansion, improving the quality of recipients, strengthening the diagnosis and treatment of rejection, and preventing cardiac allograft vasculopathy were reviewed, aiming to prolong the survival and enhance the quality of life of patients with end-stage heart failure on the waiting list or underwent heart transplantation.

The clinical value of fluorescence quantitative assay and G6PD/6PGD ratio method in the diagnosis of glucose-6-phosphate dehydrogenase deficiency and gene mutation / 中华检验医学杂志

Objective:To investigate the diagnostic value of fluorescence quantitative method and G6PD/6PGD ratio method in glucose-6-phosphate dehydrogenase (G6PD) deficiency and the type of gene mutation.Methods:A total of 1 201 patients (711 males and 490 females) with suspected G6PD deficiency in Shanghai Children′s Hospital were collected from June 2018 to March 2021. Fluorescence quantification method, G6PD/6PGD ratio method and multicolor melting curve were used to detects enzyme activity, ratio and gene mutation type. Comparison of each index and evaluation of its diagnostic efficiency were performed.Results:Among 1 201 suspicious samples, 163 cases (135 males and 28 females) were finally diagnosed. 156 cases were diagnosed by fluorescence quantitative method with a detection rate of 95.71%, and 140 cases were diagnosed by G6PD/6PGD ratio method with a detection rate of 85.89%. enzymatic activity of G6PD and ratio of G6PD/6PGD in male were significantly lower than female, and the differences were statistically significant ( U=642.5, 734.5, P<0.001). 112 cases received G6PD gene mutation detection and 92 cases were diagnosed, 74 were hemizygous mutations, 1 were homozygous mutations, 15 were heterozygous mutations, and 2 were compound heterozygous mutations. Among 15 cases of heterozygous mutations, 11 cases were diagnosed by fluorescence quantitative method, the diagnosed rate was 73.33%, 4 cases were diagnosed by G6PD/6PGD ratio method, and the diagnosed rate was 26.67%. A total of 7 mutation sites were detected and the proportions were c.1388G>A (32.22%), c.1376G>T (30.00%), c.871G>A (13.33%), c.1024C>T (11.11%). c.95A>G (7.78%), c.487G>A (4.44%), c.392G>T (1.11%). The enzymatic activities of c.1376G>T and c.1024C>T, c.487G>A were statistically significant ( P<0.001,0.015); the G6PD/6PGD ratios of c.1024C>T and c.1388G>A, c.1376G>T were statistically significant ( P=0.017,0.002,0.011,0.013). Fluorescence quantitative method had sensitivity of 100%, specificity of 95.65%, and the area under the curve (AUC) is 0.972. The sensitivity of the G6PD/6PGD ratio method was 100%, the specificity was 94.57%, and the AUC was 0.979. The sensitivity of fluorescence quantitative method combined with G6PD/6PGD ratio was 96.7%, the specificity was 100%, and the AUC was 0.992. Conclusions:Compared with fluorescence quantification, the G6PD/6PGD ratio method might not be able to diagnose female heterozygotes effectively; The panel of G6PD fluorescence quantification and G6PD/6PGD ratio was helpful to reduce the missed diagnosis. Combined with gene mutation analysis, it could improve the diagnosis rate of G6PD deficiency in the children.

Risk factors analysis of central airway stenosis after lung transplantation / 器官移植

Objective To investigate the risk factors of central airway stenosis after lung transplantation. Methods Clinical data of 155 recipients undergoing lung transplantation in Wuxi People's Hospital Affiliated to Nanjing Medical University from July 2016 to December 2017 were retrospectively analyzed. According to the incidence of central airway stenosis following lung transplantation, all recipients were divided into the stenosis group (n=36) and control group (n=119). The incidence of central airway stenosis after lung transplantation was summarized. The risk factors of central airway stenosis after lung transplantation were assessed by univariate and multivariate logistic regression analyses. Results Among 155 lung transplant recipients, 36 cases (23.2%) developed central airway stenosis. The average incidence time was (53±13) d after lung transplantation. Univariate analysis demonstrated that bilateral lung transplantation, grade 3 primary graft dysfunction (PGD), airway fungal infection, long cold ischemia time, long mechanical ventilation time and long intensive care unit (ICU) stay were the risk factors for central airway stenosis after lung transplantation (all P < 0.05). Multivariate analysis showed that airway fungal infection, long cold ischemia time and long mechanical ventilation time were the independent risk factors for central airway stenosis after lung transplantation (all P < 0.05). Conclusions Airway fungal infection after lung transplantation, long cold ischemia time and long mechanical ventilation time probably lead to central airway stenosis after lung transplantation. Active preventive measures and intimate monitoring should be taken to improve the quality of life of the recipients after lung transplantation.

Dihydrolipoamide Acetyltransferase Promotes Nucleic Acid Synthesis by Controlling Phosphogluconate Dehydrogenase Acetylation / 中国生物化学与分子生物学报

Mitochondrial pyruvate dehydrogenase complex (PDC) is crucial for glucose homeostasis in mammalian cells‚ decarboxylation of glycolytic intermediate pyruvate to acetyl coenzyme A (acetyl-CoA) in mitochondria. Dihydrolipoyl acetyltransferase (DLAT) is a subunit of the pyruvate dehydrogenase complex. Here‚ we reported that DLAT was commonly increased in lung cancer and its expression was associated with worse clinical outcomes. We found that suppression of DLAT in lung cancer cells resulted in reduced nucleic acid biosynthesis and attenuated cancer cell proliferation through controlling acetylation level of 6-phosphogluconate dehydrogenase (6PGD) ‚ the third enzyme in the oxidative pentose phosphate pathway (PPP) ‚ in which ribulose-5-phosphate (Ru-5-P) is produced for nucleic acid biosynthesis. Together‚ our study contributes to recent interest and discussion cross talk in cancer metabolism‚ which contributes to tumor growth. Future mechanistic studies should lead to the elucidation of the mode of action of DLAT in human lung cancer and establish DLAT as a viable drug target.

Effect of hydroxysafflor yellow A on COX-2/PGD2/DPs pathway in cortex of mice with cerebral ischemia-reperfusion injury / 中国药理学通报

Aim To observe the effect of hydroxysafflor yellow A (HSYA) on the COX-2/PGD/DPs pathway in the cortex of mice with cerebral ischemia-reperfusion injury. Methods C57BL/6 male mice were randomly divided into sham group, model group and HSYA group. Right middle cerebral artery occlusion/reperfusion model was established in mice by intraluminal suture method. HYSA (20 mg • kg"1) was injected into the tail vein for five consecutive days before the operation. The sham group and the model group were given the same volume of normal saline. The neurological function score and cerebral infarct volume were measured 24 hours after operation. The histopathological changes of mouse cortex were observed by HE staining. The protein and mRNA expression of COX-2, DP, and DP2 were detected by Western blot and qRT- PCR respectively. The levels of TNF-a, IL-1 (3 and PGD2 were detected by ELISA. Results Compared with sham group, the scores of neurological function, infarct volume, the expression of COX-2, DP, and DP2 protein and mRNA, and the contents of TNF-a, IL-1 (3 and PGD2 in the cortex of model group significantly increased. Compared with model group, the scores of neurological function and the infarct volume significantly decreased in HSYA-treated group, and the damage of cortical cells in ischemic area was significantly improved. The expressions of COX-2, DP, and DP2mRNA and protein were significantly down-regula- ted, and the levels of inflammatory factors such as TNF-a, IL-1 p and PGD2 were markedly down-regula- ted. Conclusions HSYA inhibits the activation of COX-2/PGD2/DPs pathway in mouse brain tissues, which may be involved in the protective mechanism of HSYA in cerebral ischemia-reperfusion injury.

Application progress of extracorporeal membrane oxygenation in the perioperative period of lung transplantation / 器官移植

Lung transplantation is the ultimate treatment for many kinds of end-stage lung diseases. However, the perioperative management of lung transplantation is complicated with high fatality of patients. Extracorporeal membrane oxygenation (ECMO) is an effective method of extracorporeal respiration and circulation support. ECMO plays an important role in the perioperative support treatment of lung transplantation, which breaks the limitation of contraindications and promotes the development of lung transplantation. In this article, the indications, catheter placement strategies and application of ECMO in the perioperative period of lung transplantation were reviewed.

Clinical analysis of lung transplantation from donors combined with pulmonary contusion / 器官移植

Objective To explore the efficacy and prognosis of lung transplantation from donors combined with pulmonary contusion on the treatment of patients with end-stage lung disease. Methods Clinical data of 73 cases of donors and recipients were collected. The donors and recipients were divided into contusion group (23 cases of donors and recipients, with a maximum diameter of contusion in 5-8 cm) and standard group (50 cases of donors and recipients) depending on combined pulmonary contusion. Major clinical indicators [postoperative oxygenation index, duration of mechanical ventilation and chest tube drainage and incidence of primary graft dysfunction (PGD)] and prognosis of the recipients in both groups were compared. Results The recipients in both groups presented no significant difference in postoperative oxygenation index, duration of mechanical ventilation and chest tube drainage and incidence of PGD (all P>0.05). The postoperative 1-year survival of the recipients in standard group and contusion group was 74％ and 83％, which presented no statistically significant difference (P>0.05). Conclusions The efficacy and prognosis of lung transplantation from donors combined with pulmonary contusion (with a maximum diameter of 5-8 cm) are comparable to those of lung transplantation from standard donors.

Rapid preimplantation genetic diagnosis of -thalassemia SEA deletion with blastocyst cell whole genome amplification and short fragment Gap-PCR method / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a rapid preimplantation genetic diagnosis method for -thalassemia SEA deletion based on blastocyst cell whole genome amplification (WGA) combined with short fragment Gap-PCR.</p><p><b>METHODS</b>Using multiple displacement amplification (MDA) WGA technique, we established a double-fluorescent PCR system of the housekeeping genes GAPDH and β-actin for WGA quality testing, and a genotyping PCR system of mutant and normal short sequences for α-thalassemia SEA deletion. The sensitivity and accuracy of this method for diagnosis of -thalassemia SEA deletion were evaluated by detecting lymphocyte samples containing different cell numbers from carriers of SEA deletion. The applicability of this method was evaluated by testing of 12 blastocyst biopsy samples.</p><p><b>RESULTS</b>Detection of lymphocyte samples with different cell numbers using the method developed in this study revealed no ADO in 3-cell samples, and the product quantity of WGA became stable for 4-cell samples. Genotyping of the 10 blastocyst biopsy samples with successful WGA showed a genotype of --/ in 5 samples and / in the other 5 samples, which were consistent with the verification results.</p><p><b>CONCLUSIONS</b>The method developed in this study is a complete testing process for 4-6 blastocyst biopsy cells to allow rapid, accurate, and cost-effective PGD genotyping of -thalassemia SEA deletion using short fragment gap-PCR.</p>

Revisão das técnicas de biologia molecular aplicadas no diagnóstico genético pré­implantacional e uma reflexão ética / Review of molecular biology techniques used in preimplantation genetic diagnosis and ethical reflection

O diagnóstico genético pré-implantacional (PGD) é uma ferramenta que permite a selec¸ão do embrião saudável por meio da análise gênica e cromossômica. Beneficia casais no grupo de risco, como, por exemplo, casais com histórico de aborto de repetição ou com histórico familiar de doença hereditária. Diante do avanço das metodologias empregadas no PGD, esta revisão tem como objetivo reunir as informações acerca das técnicas de biópsia, de biologia molecular e aspectos bioéticos dessa prática. Assim, foi possível agregar as informações como vantagens, desvantagens, restrições e indicações referentes ao estágio embrionário em que é retirado o material genético e as técnicas de biologia molecular usadas na análise genética. Além disso, foi possível especificar alguns questionamentos bioéticos que surgem com a prática do PGD, como, por exemplo, a possível eugenia. Concluiu-se que a análise da trofoectoderme dos embriões e da aplicação da tecnologia de NGS é promissora para o futuro do PGD, bem como a primordialidade da criação de leis que regem e completam as lacunas no sentido ético e moral dessa prática.(AU)

The preimplantation genetic diagnosis (PGD) is a tool that allows the selection of healthy embrios through gene and chromosome analysis, benefiting couples at risk. Like couples with recurrent miscarriage or family history of hereditary disease. Given the advance of the methodologies used in PGD this review aims to assemble information about the biopsy techniques, molecular biology and bioethical aspects of this practice. Therefore, it was possible collect the information like advantages, disadvantages, restrictions and indications referring to embryonic stage in which is made the removal of genetic material and molecular biology techniques used in genetic analysis. In addition, some bioethic questions that arise with the practice of PGD were verified as, for example, possible Eugenia. It was concluded that the analysis of trofoectoderme of embryos and the application of NGS technology is promising for the future of PGD, as well as the primordiality the creating laws governing and complete the gaps in the ethical and moral sense of this practice.(AU)

Preimplantation Genetic Diagnosis of X-linked Severe Combined Immunodeficiency / 中山大学学报(医学科学版)

[Objective]To establish a reliable and accurate preimplantation genetic diagnosis (PGD)method using multiple dis?placement amplification (MDA), which can be applied to the diagnosis of X-linked severe combined immunodeficiency disease (X-SCID).[Methods]Haplotype analysis for the X-SCID family was performedusing five short tandem repeats (STR) markers flanking the both sides of the interleukin-2 (IL-2) receptor gamma chain (IL2RG) gene. MDA technique was used for single-cell whole genomic amplification. The products were used as template in polymerase chain reaction (PCR) of informative STR markers found by linkage analysis for haplotype analysis as well as sequencing of the IL2RG gene exon 5.The amelogenin (AMEL) locus was used to do sex diag?nosis.[Results]Linked analysis revealed 3 STR markers were informative. The method was evaluated with 10 single lymphocytes and 10 single blastomeres. MDA was successful in all single cell. The detection efficiency of gene sequencing of pathogenic IL 2RG exon5 was 100%. The PCR efficiency of 3 STR informative markers and AMEL was 96.3%(77/80)and the average allele drop-out (ADO) rate was 11.5%(7/61). A cycle of PGD was performed on the family, and seven embryos were diagnosed, two of which were normal embry?os. Twin pregnancy occurred after transplantation which were given a healthy baby boy and a healthy baby girl.[Conclusion]In this study, multiple displacement amplification combined with specific amplification/sequencing of pathogenic gene and haplotype analysis in the single cell level of X-linked severe combined immunodeficiency disease were performed. The protocol can avoid misdiagnosis caused by contamination and ADO, and improve the diagnostic efficiency of PGD.

Possible role of PGD2 in malaria infections

OBJECTIVE@#To preliminarily investigate the possible role of prostaglandin D2 (PGD2) in malaria infections.@*METHODS@#Blood and urinary samples (n = 120 each) were collected from Thai patients with Plasmodium falciparum (P. falciparum) with moderate (n = 26) and high (n = 4) parasitemia, patients with Plasmodium vivax (P. vivax) (n = 30), patients with fever associated with other infections (n = 30), and healthy subjects (n = 30). PGD2 concentrations in plasma and urinary samples of healthy subjects, patients with fever associated with other infections and patients with malaria were determined using Prostaglandin D2-MOX express EIA kit (Cayman Chemical, USA).@*RESULTS@#The possible association between PGD2 and malaria infections is clearly demonstrated with PGD2 concentration in urine. The urinary PGD2 concentrations were relatively high (about 5-fold) in patients with P. falciparum with moderate parasitemia and P. vivax infections compared with other groups. Furthermore, the concentration in patients with P. falciparum with moderate parasitemia and P. vivax infection were significantly higher than that in healthy subjects and patients with fever associated with other infections.@*CONCLUSIONS@#Urinary PGD2 concentrations may offer a more dependable and useful tool for predicting malaria severity. Confirmation is this preliminary finding is required with a larger sample size.

Effects of DP2 intervention on primary cultured rat hippocampal neuron treated with aluminum overload / 中国药理学通报

Aim To establish primary cultured rat hip-pocampal neuron damage model induced by aluminum maltolate and study the effect of intervention for DP2 on primary cultured rat hippocampal neuron treated with aluminum overload. Methods The hippocampus was dissected out from fetal rat ( embryonic 18 d ) . After being cultured for 7 d, the hippocampal neuron was treated with Al( malt) 3 to establish the model of prima-ry cultured rat hippocampal neuron damage and mean-while treated with DP2 agonist DK-PGD2 and DP2 an-tagonist CAY10471, respectively. After treatment for 24 h, the cell viability was measured by MTT and LDH, Ca2+ fluorescence intensity. Neuronal pathomor-phology was observed by HE staining. Results The purity of hippocampal neuron was more than 95%. Compared with the control group, the number of hipp-ocampal neurons was reduced and neurons became chromatic agglutination and karyopyknosis in aluminum overload group. Treatment of aluminum caused a sig-nificant decrease in MTT value ( P<0. 01 ) and an in-crease in the LDH leakage rate (P<0. 01). The Ca2+fluorescence intensity significantly increased ( P <0. 01 ) in aluminum overload group. Compared with that of the aluminum overload group, treatment of DK-PGD2 , a selective DP2 agonist, significantly aggravated the primary cultured rat hippocampal neuron injury caused by aluminum overload accompanied with the significant decrease of MTT value ( P <0. 01 , P <0. 05 ) and an increase of the LDH leakage rate ( P<0. 01), significant increase of Ca2+ fluorescence inten-sity of neuron. Treatment of CAY10471, a selective DP2 antagonist, had opposite effects of DK-PGD2 . Conclusion The activation of DP2 can increase hipp-ocampal neural susceptibility to aluminum overload.

Assisted Reproduction and Cross-Border Maternal Surrogacy Regulations in Selected Nations.

Aims: To ascertain the laws and policies of selected high income countries, with respect to the disposition of their citizens seeking assisted reproductive technologies (ARTs) internationally. Study Design: Literature review. Methodology: PubMed, Scopus and Google of various ART terms with terms relating to regulations in the selected nations of Australia, Canada, New Zealand, the UK, the USA, and Israel. Results: All nations except the USA have a federal ART regulatory presence, distinguish between gestational and traditional surrogacy, and between paid and unpaid surrogacy. Policies concerning the repatriation of children produced by ART abroad vary widely. Conclusions: Heterogeneous regulations are one of the drivers of the global reproductive tourism industry. Domestic regulations are likely affected by both the values of a specific population and the needs of the industry.

The role of the airway remodeling in chronic obstructive pulmonary disease PGD 2 regulate TGF-β1 / 重庆医学

Objective To explore PGD2 biological characteristics in L-929 mice lung fibroblasts cell by Laropiprant is a specific in-hibitor of PGD2 receptor regulation of TGF-β1/Smads signaling pathway .Provide further basis research to explore the molecular mechanisms of airway fibrosis .Methods The cells divided by Laropiprant concentration 0 .3 μmoL group ,1 .0 μmoL group ,3 .0μmoL group ,10 .0 μmoL group ,30 .0 μmoL and the control group .Each group were added TGF-β2 (2 .5 ng/mL) were cultured for 24 h after stimulation with the corresponding concentrations of Laropiprant 24 h ,TGF-β1 Smad3 and Smad4 expression were detected by PCR and WB ,respectively .A randomized approach ,using different concentrations of Laropiprant acts on cells at different times (12 ,24 ,48 ,72 h) by MTT assay for cell growth Laropiprant inhibition .Results TGF-β1 ,Smad3 and Smad4 mRNA expression de-creased with the addition of Laropiprant concentration increases ,there was significant difference compared with the control group with (P<0 .05) .TGF-β1 ,Smad3 and Smad4 protein expression decreased with the addition of Laropiprant concentration increases ,there was significant difference compared with the control group with (P< 0 .05) .Cell growth inhibition rate decreased with Laropiprant concentration increasing and cell culture time growth ,Laropiprant in concentration 1μml ,reaction time was 24 to 36 h ,the cell growth inhibition was significantly improved .Conclusion PGD2-DP1 expression in L-929 mouse lung fibroblasts cell may be associated with TGF-β1/Smads regulation .growth inhibition rate decreased with Laropiprant concentration increasing and cell culture time growth ,re-action time prolong ,the cell growth inhibition was significantly improved .

Atención hospitalaria en la vigilancia obstétrica y metabólica de la gestante diabética, una experiencia de trabajo / Hospital attention in obstetric and metabolic surveillance in diabetic pregnant women; a work experience

Introducción: se conoce que en los últimos años, el manejo obstétrico ha enfatizado el control estricto de la glicemia en la madre y que ha mejorado la sobrevida fetal, la cual es directamente proporcional a la glicemia media materna. Objetivo: caracterizar los principales resultados en la experiencia hospitalaria sobre la vigilancia obstétrica y metabólica en la atención de gestantes diabéticas en el Hospital General Ciro Redondo García, Centro de Referencia Territorial en Artemisa. Métodos: se realizó un estudio observacional analítico, prospectivo y de corte longitudinal en el Hospital General Docente Ciro Redondo García de Artemisa desde junio de 2005 hasta junio de 2012. De un universo de 2 140 gestantes ingresadas, 240 fueron diagnosticadas diabéticas, constituyendo la muestra de estudio, Resultados: como diabéticas gestacionales (DG) se clasificó el 77,5 por ciento mientras que las diabéticas pregestacionales (DPG) constituyeron el 22,5 por ciento .Los grupos de edades de 31 a 36 años y de 20 a 25 años fueron los de mayor predominio en la diabetes gestacional pregestacional para un 29,1 por ciento y un 33,3 por ciento respectivamente. Los factores de riesgo de mayor predominio en el estudio fueron: la obesidad (44,2 por ciento ), la edad mayor de 34 años, polihidramnios, macrosomía previa y los abortos espontáneos (38,7; 18; 8,1; y 6,6 por ciento respectivamente). Otros factores de riesgo fueron los antecedentes familiares de diabetes mellitus de las gestantes, el diagnóstico de la diabetes gestacional después de las 20 semanas, la cesárea como el tipo de parto de mayor predominio y la edad gestacional a término al parto. Conclusiones: en un valorado sistema de salud pública como el cubano, se debe tomar medidas pertinentes para monitorear y controlar la morbilidad y complicaciones de las gestantes diabéticas

Background: it is known that, in recent years, obstetric management has made emphasis on the strict control of glycemia in the mother and the fetal survival has been improved, which is directly proportional to the mean maternal glycemia. Objective: to characterize the principal results in the hospital experience on obstetric and metabolic surveillance in the management of pregnant diabetic women in Ciro Redondo García General Hospital, Regional Reference Center in Artemisa. Methods: an observational analytical prospective cross-longitudinal study was conducted in Ciro Redondo García General Hospital in Artemisa from June 2005 to June 2012. From a universe of 2 140 pregnant women admitted in the hospital, 240 were diagnosed diabetics, representing them, the sample of the study. Results: the 77.5 percent of them were classified as gestational diabetic women (GD) whereas pregestational diabetic women (PGD) represented the 22.5 percent .The 31-36 and 20-25 age groups were the most predominant in gestational and pregestational diabetes, for a 29.1 percent and a 33.3 percent, respectively. The most predominant risk factors in the study were: obesity (44.2 percent ), age over 34 years, polyhydramnios, previous macrosomia, and spontaneous abortion (38.7, 18, 8.1, 6.6 percent respectively). Other risk factors were: family antecedents of diabetes mellitus in pregnant women, the time of diagnosis of gestational diabetes after the 20 weeks, the caesarean section as the most predominant type of delivery, and gestational age at delivery at term. Conclusions: in a valued Public Health System such as the Cuban one, appropriate measures should be taken to monitor and control morbidity and complications in pregnant diabetic women

Desenvolvimento e validação de um método por CLAE-EM/EM para determinação de PGE2 e PGD2 em meio de cultivo celular e avaliação da recuperação das prostaglandinas utilizando diferentes condições na extração em fase sólida / Development and validation of HPLC-MS/MS method to determine PGE2 and PGD2 in cell culture medium and assessment of recovery of the prostaglandins by solid phase extraction under various conditions

Tradicionalmente, estas prostaglandinas são quantificadas por técnicas de imuno-ensaio, que apresentam diversas desvantagens. Estes metabólitos são isômeros estruturais, e dessa forma é necessário o uso de técnicas de detecção seletivas, como cromatografia líquida acoplada à espectrometria de massas sequencial (CLAE-EM/EM). Para a extração de prostaglandinas de matrizes complexas, destaca-se a extração em fase sólida (EFS), que otimizada, fornece excelentes taxas de recuperação. O objetivo deste trabalho foi desenvolver e validar um método rápido por CLAE-EM/EM, para análise simultânea de PGE2 e PGD2 de meio de cultivo celular e avaliar a eficiência de extração em diferentes condições de EFS, em relação ao método proposto pelo fabricante dos cartuchos. A separação ocorreu com coluna de fase reversa (C18, 150mm x 2.1mm, 5μm) eluída no modo gradiente com acetonitrila e água (0,1% AFO). Dez condições diferentes de EFS foram testadas. O método desenvolvido foi adequado para a análise simultânea de PGE2 e PGD2 , apresentando resolução de ~1,5 entre os picos e corrida de 11 minutos. LD da ordem de 0,5 ng/mL e LQ de 1,0 ng/mL foram obtidos para ambos os analitos. A linearidade de PGE2 e PGD2 apresentou r>0,99. Variações inferiores a 6,51% e 5,93% foram encontradas para repetibilidade e precisão intermediária, respectivamente. Foi possível diminuir perdas durante a EFS e aumentar a recuperação dos analitos. A condição que ofereceu melhor eficiência de extração aumentou o rendimento em 181% para PGE2 e 323% para PGD2 , em relação ao método proposto pelo fabricante.

PGE2 and PGD2 are very important pro-inflammatory mediators. Traditionally, these prostaglandins are estimated by immunoassay techniques, which have several disadvantages. Since these metabolites are structural isomers, it is necessary to use selective detection techniques, such as liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). For the extraction of prostaglandins from complex matrices, solid phase extraction (SPE) is an outstanding method that can be optimized to provide excellent recovery. The aim of this study was to develop and validate a rapid method for the simultaneous analysis of PGE2 and PGD2 in cell culture medium by HPLC-MS/MS and to assess the extraction efficiency of SPE under various conditions, compared to the generic method proposed by the manufacturer of the cartridges. The analytes were separated on a reversed-phase column (C18, 150mm x 2.1mm, 5μm), eluted in a gradient of acetonitrile and water (0.1% formic acid). Ten different conditions for SPE were tested. The method was suitable for the simultaneous analysis of PGE2 and PGD2 , showing a resolution of ~1.5 between the peaks and a run time of 11 minutes. LOD of 0.5 ng/mL and LOQ of 1.0 ng/mL were recorded for both analytes. The linearity of the analytical curves for both PGE2 and PGD2 showed r>0.99. Variations of less than 6.51% and 5.93% were found for repeatability and intermediate precision, respectively. It was possible to reduce the losses during SPE and enhance the recovery of the analytes. The condition affording the best extraction efficiency increased the yield by 181% for PGE2 and 323% for PGD2 , relative to the method proposed by the manufacturer.

Diagnóstico genético preimplantacional: análisis de aneuploidías únicas / Preimplantational genetic diagnosis: single aneuploidies analysis

Introducción: De las causas más conocidas en cuanto a la falta del éxito en el embarazo con tratamientos de reproducción asistida son aquellas relacionadas a las aneuploidías cromosómicas presentes en los embriones. El diagnóstico genético preimplantacional (PGD) es una técnica empleada en reproducción asistida para detectar estas anomalías, seleccionando aquellos que sean cromosómicamente normales, para luego transferirlos al útero de la paciente. Los embriones con aneuploidías únicas podrían tener la capacidad de sobrevivir y lograr la implantación, y por lo tanto, sin diagnóstico previo, estas podrían pasar desapercibidas. Objetivos: Determinar la incidencia de aneuploidías únicas en embriones de buena calidad embrionaria en el día 3 de desarrollo hasta blastocisto. Diseño: Estadístico y experimental. Instituciones: Reprogenetics Latinoamérica y Centro de Reproducción asistida, de la Clínica Concebir. Material Biológico: Muestras de biopsia embrionaria. Metodología: Análisis comparativo de resultados a partir de la evaluación de cada muestra obtenida por biopsia en el día tercero y día quinto de desarrollo embrionario, realizando el PGD por hibridación in situ (FISH) y genómica comparada (aCGH), respectivamente. Resultados: El 62,9% de embriones que presentaron monosomías únicas al tercer día de desarrollo embrionario resultaron ser de 8 células. Pero cuando se evaluó por aCGH en día cinco, 42,3% resultó anormal, y de estos 37,5% perteneció al estadio de 8 células. El índice de monosomías únicas en blastocisto resultó ser 57,9% de un total de 84,2% de aneuploidías únicas. Conclusiones: Los embriones de 8 células en el tercer día de desarrollo embrionario son los más probables de llegar al estadio de blastocisto, así como presentar aneuploidías únicas.

Background: Known causes of unsuccessful pregnancy in couples undergoing assisted reproduction treatment include embryo aneuploidies. Preimplantation genetic diagnosis (PGD) is a technique used in assisted reproduction in order to detect these abnormalities, select embryos chromosomally normal and subsequently transfer to the patients’ uterus. Embryos with single aneuploidies may have the ability to survive and achieve unnoticed implantation. Objectives: To determine incidence of single aneuploidies in good quality embryos in third day of development to blastocyst. Design: Statistical and experimental study. Setting: Reprogenetics Latin-America and Assisted Reproduction Center - Concebir. Biologic material: Samples of embryo biopsies. Methods: Comparative analysis of results from evaluation of each sample obtained by embryo biopsy on the third and fifth days of embryonic development, performing PGD by respectively in situ hybridization (FISH) and comparative genomics (aCGH). Results: On third day of embryonic development 62.9% of embryos with single monosomy had 8-cell morphology. Though when evaluated by aCGH in the blastocyst stage 42.3% were abnormal and 37.5% of these belonged to the 8-cell stage. Single monosomies index in the blastocyst stage was 57.9% in 84.2% of single aneuploidies. Conclusions: Eight-cell embryos on the third day of embryonic development are most likely to reach blastocyst stage and have single aneuploidies.

Técnicas emergentes en medicina reproductiva: diagnóstico cromosómico del primer corpúsculo polar del ovocito / Emerging techniques in reproductive medicine: chromosomal diagnosis of the oocyte first polar body

El Diagnóstico Genético Preimplantacional (PGD) se ha convertido en una herramienta de rutina para la detección de anormalidades cromosómicas o genéticas, en muchos países del mundo. Se han reportado más de 20.000 ciclosde PGD, desde su desarrollo hace más de 20 años, habiendo nacido más de 4.000 niños hasta el año 2007. En Chile, esta técnica es realizada por la Unidad de Medicina Reproductiva de Clínica Las Condes, y se realiza sólo en la variante previa a la fecundación, en donde se biopsia el primer corpúsculo polar y sólo se insemina a los ovocitos encontrados cromosómicamente sanos. Las indicaciones más comunes para este tratamiento son: 1) evitar el aborto en pacientes con aborto recurrente sin explicación anatómica ni clínica; 2) mejorar las tasas de implantación en mujeres mayores de 37 años con antecedentesde procedimientos anteriores en los que se transfirieron embriones de buena calidad; 3) evitar el nacimiento deniños con enfermedades de origen cromosómico en mujeres mayores de 39 años.

Pre-implantational Genetic Diagnosis has become a common tool in most countries of the world. In almost 20 years since its development, it has been reported more than 20,000 cycles of PGD and till 2007, more than 4,000 children have been born. In Chile, this technique is done by the Unit of Reproductive Medicine of Clínica Las Condes. It is done only in the mode previous to fertilization. In where we study polar bodies and only chromosomically healthy oocytes are inseminated. The most common indications for this treatment are: 1) to avoid abortions in patients with recurrent abortion without anatomical nor clinical explanation; 2) to improve implantation rates in women older than 37 years of age, with previous procedures in which good quality embryos were transferred; 3) to avoid birth of children with diseases of chromosomal origin in women over 39 year of age.

A Study of Guidelines for Genetic Counseling in Preimplantation Genetic Diagnosis (PGD)

PURPOSE: Preimplantation genetic diagnosis (PGD), also known as embryo screening, is a pre-pregnancy technique used to identify genetic defects in embryos created through in vitro fertilization. PGD is considered a means of prenatal diagnosis of genetic abnormalities. PGD is used when one or both genetic parents has a known genetic abnormality; testing is performed on an embryo to determine if it also carries the genetic abnormality. The main advantage of PGD is the avoidance of selective pregnancy termination as it imparts a high likelihood that the baby will be free of the disease under consideration. The application of PGD to genetic practices, reproductive medicine, and genetic counseling is becoming the key component of fertility practice because of the need to develop a custom PGD design for each couple. MATERIALS AND METHODS: In this study, a survey on the contents of genetic counseling in PGD was carried out via direct contact or e-mail with the patients and specialists who had experienced PGD during the three months from February to April 2010. RESULTS: A total of 91 persons including 60 patients, 49 of whom had a chromosomal disorder and 11 of whom had a single gene disorder, and 31 PGD specialists responded to the survey. Analysis of the survey results revealed that all respondents were well aware of the importance of genetic counseling in all steps of PGD including planning, operation, and follow-up. The patient group responded that the possibility of unexpected results (51.7%), genetic risk assessment and recurrence risk (46.7%), the reproduction options (46.7%), the procedure and limitation of PGD (43.3%) and the information of PGD technology (35.0%) should be included as a genetic counseling information. In detail, 51.7% of patients wanted to be counseled for the possibility of unexpected results and the recurrence risk, while 46.7% wanted to know their reproduction options (46.7%). Approximately 96.7% of specialists replied that a non-M.D. genetic counselor is necessary for effective and systematic genetic counseling in PGD because it is difficult for physicians to offer satisfying information to patients due to lack of counseling time and specific knowledge of the disorders. CONCLUSIONS: The information from the survey provides important insight into the overall present situation of genetic counseling for PGD in Korea. The survey results demonstrated that there is a general awareness that genetic counseling is essential for PGD, suggesting that appropriate genetic counseling may play a important role in the success of PGD. The establishment of genetic counseling guidelines for PGD may contribute to better planning and management strategies for PGD.