Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production / 药学学报

Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

Correlation between neonatal retinal hemorrhage and changes in umbilical artery blood gas analysis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the correlation between neonatal retinal hemorrhage(RH)and changes in umbilical artery blood gas analysis.METHODS: A total of 312 full-term neonates born in our obstetrics department from January 2019 to December 2021 were selected as the study subjects. According to the RetCam III fundus examination results, 245 neonates who did not experience RH were included in the control group, while 67 cases with RH were found to be included in the RH group. In addition, neonates were grouped into I degree group(n=20), II degree group(n=29), and III degree group(n=18)based on the degree of RH. General clinical data and umbilical artery blood gas analysis indicators between the RH group and the control group were compared; the levels of umbilical artery blood gas analysis indicators in neonates with different degrees of RH, the relationship between pH and RH degree, and the influencing factors of neonatal RH were analyzed.RESULTS: There was no obvious difference in maternal age, average gestational week, fetal gender, parity, gestational diabetes, fetal birth weight, and amniotic fluid between the RH group and the control group(all P>0.05), while there were obvious differences in delivery methods, gestational hypertension, forceps assisted delivery, neonatal asphyxia, and umbilical cord around the neck(all P<0.05). The pH value, arterial blood sample partial pressure(PaO2)and base excess(BE)values of the RH group were obviously lower than those of the control group(all P<0.01), while the arterial carbon dioxide partial pressure(PaCO2)was obviously higher than that of the control group(P<0.01). There were obvious differences in umbilical artery blood gas analysis indicators among children with different degrees of RH(P<0.05), and with the increase of the degree of RH, pH value, PaO2 and BE gradually decreased(P<0.05), and PaCO2 gradually increased(P<0.05). There was a negative correlation between the degree of RH and the pH of umbilical artery blood gas analysis(rs=-0.593, P<0.05). The results of multivariate Logistic regression analysis showed that delivery method, gestational hypertension, forceps assisted delivery, neonatal asphyxia, umbilical cord entanglement, pH, PaO2, PaCO2, and BE were all influencing factors for the occurrence of neonatal RH.CONCLUSION: There is a close correlation between neonatal RH and changes in umbilical artery blood gas analysis, and umbilical artery blood gas analysis can be used for the diagnosis of neonatal RH, which can be used to guide clinical treatment.

In Vitro and in Vivo Evaluation of Scutellarin-phospholipid Complex Nanoemulsion and Analysis of Its Activity in Ameliorating LPS-induced Vascular Endothelial Injury / 中国实验方剂学杂志

ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion（SCU-PC-NE）， such as release， cell uptake and tissue distribution， and to investigate its effect on ameliorating lipopolysaccharide（LPS）-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC， ethyl oleate， Kolliphor HS15， 1，2-propylene glycol（50， 400， 514.3， 85.7 mg）， respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope， the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis， thiazolyl blue（MTT） colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells（HUVECs）， the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe， the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS（1 mg·L-1） and HUVECs， the effect of SCU-PC-NE on the levels of interleukin（IL）-1β and IL-6 were determined by enzyme-linked immunosorbent assay（ELISA）， 18 Kunming male mice were randomly divided into blank group， model group， blank preparation group（equivalent to high dose group）， SCU group and SCU-PC-NE low and high dose groups（5， 10 mg·kg-1）， 3 mice in each group， and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h， equal volume of normal saline was given to the blank group and the model group， and the drug was administered for 4 consecutive times. Except for the blank group， the endothelial inflammatory injury was induced by intraperitoneal injection of LPS（10 mg·kg-1） at 12 h before the last administration in each group. Hematoxylin-eosin（HE） staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light， mostly spherical with rounded appearance and relatively uniform particle size distribution， with the average particle size of 35.31 nm， Zeta potential of 7.23 mV， and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group（P<0.01）. Compared with normal mice， the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen， kidney， brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS（P<0.05， P<0.01）， especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group（P<0.05， P<0.01）， and compare with the model group， all administration groups significantly down-regulated IL-1β level， with the strongest effect in the SCU-PC-NE high-dose group（P<0.01）， and all administration groups significantly down-regulated IL-6 level， with the strongest effect in the SCU-PC-NE low-dose group（P<0.05）. Compare with the blank group， the results of HE staining showed that the endothelial cells were damaged， the elastic fibers were broken and arranged loosely in the model group， although similar vascular injury could be observed in the blank preparation group， SCU group and SCU-PC-NE low-dose group， the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE， which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS， suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.

Inhibition of HSP70 release by geniposide improves angiogenesis in moist heat arthralgia spasm syndrome collagen induced arthritis rats / 中国药理学通报

Aim To investigate the relation between the effect of geniposide (GE) in improving angiogenesis in arthralgia spasm syndrome collagen induced arthritis (CIA) rats and the modulation of heat shock proteins 70 (HSP70) release. Methods A CIA model was constructed by multiple intradermal injections of complete Freund's adjuvant (CFA) and an equal volume mixture of chicken type II collagen (CCII) into the dorsal and caudal root regions of rats, on the basis of which a rheumatic fever stimulus was given to build up a moist heat arthralgia spasm syndrome in CIA rats. After successful modeling, the groups were randomly grouped, and the administered groups were gavaged with GE (60, 120 mg · kg

Effects of inhibiting CAMKⅡ expression in ARPE-19 cells on the migration, invasion, and tube formation of HUVECs in a non-contact co-culture system / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of inhibiting Ca2+/calmodulin-dependent protein kinase Ⅱ(CAMKⅡ)expression in adult retinal pigment epithelial cell line-19(ARPE-19)cells on the migration, invasion, and tube formation of human umbilical vein endothelial cells(HUVECs)in a non-contact co-culture system.METHODS: RNA sequencing was performed on ARPE-19 cells overexpressing CAMKⅡ-δ, and bioinformatics was used to analyze the biological functions of the differentially expressed genes. Transwell inserts was used to construct a non-contact co-culture system of ARPE-19 and HUVECs. The experimental groups included: blank group: only HUVECs were inoculated without ARPE-19 cells; control group: ARPE-19 and HUVECs cells were co-cultured with complete medium; AIP group(CAMKⅡ inhibition group): ARPE-19 cells in AIP(160 nmol/L)were co-cultured with HUVECs in complete medium. The migration, invasion and tube formation abilities of HUVECs were detected. The protein expression levels of CAMKⅡ/AMPK/mTOR/VEGFA were detected by Western blotting.RESULTS:Bioinformatics analysis found that the differentially expressed genes could affect biological processes such as cell growth and death and cell movement. The scratch test and transwell migration test showed that the relative mobility of HUVECs in the AIP group was significantly lower than that in the control group(all P&#x003C;0.05). However, the invasion and tube formation assay showed that the relative invasion rate and tube formation rate of the AIP group were not significantly different from those of the control group(both P&#x003E;0.05). Western blotting results showed that the expression levels of CAMKⅡ, P-mTOR, and VEGFA proteins in the AIP group were significantly lower than those in the control group, while the expression level of the P-AMPK protein was significantly higher than that in the control group(all P&#x003C;0.05).CONCLUSION:In the non-contact co-culture system, inhibition of CAMKⅡ expression in ARPE-19 cells significantly reduced the migration ability of HUVECs, but it cannot change the invasion and tube formation ability, which may be achieved by AMPK/mTOR/VEGFA.

Interpretation of 2023 American Heart Association and American Academy of Pediatrics focused update on neonatal resuscitation guidelines / 中国当代儿科杂志

In November 2023, the American Heart Association and the American Academy of Pediatrics jointly released key updates to the neonatal resuscitation guidelines based on new clinical evidence. This update serves as an important supplement to the "Neonatal resuscitation: 2020 American Heart Association guidelines for cardiopulmonary resuscitation and emergency cardiovascular care". The aim of this paper is to outline the key updates and provide guidance on umbilical cord management and the selection of positive pressure ventilation equipment and its additional interfaces in neonatal resuscitation.

Effect of Zuogui Jiangtang Tongmai Prescription on Inflammatory Injury of Human Umbilical Vein Endothelial Cells Induced by High Glucose and LPS Based on GPR43/β-arrestin-2/IκBα/NF-κB Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effects and mechanism of Zuogui Jiangtang Tongmai prescription （ZJTP） on human umbilical vein endothelial cells （HUVECs） damaged by high glucose combined with lipopolysaccharide （LPS）. MethodThe survival rate of cells was determined by cell counting kit-8 （CCK-8）， and the level of tumor necrosis factor-α （TNF-α） was determined by enzyme-linked immunosorbent assay （ELISA） to determine the optimal injury concentration and action time of LPS， as well as the optimal action concentration of ZJTP drug-containing serum. HUVECs were divided into a blank control group， a model group， a ZJTP drug-containing serum group， and an SCFA mixed liquid group. ELISA was used to detect the level of endothelin-1 （ET-1）， nitric oxide （NO）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and TNF-α. Western blot was performed to detect the protein expression of G protein-coupled receptor43 （GPR43）， β-suppressor protein-2 （β-arrestin-2）， nuclear factor-κB suppressor α （IκBα）， and nuclear factor κB p65 （NF-κB p65）. The nucleation of NF-κB p65 was observed by immunofluorescence staining （IF）. The role of GPR43 in the regulation of inflammatory injury was observed by means of small interfering ribonucleic acid （siRNA）. The cells after intervention were divided into an empty carrier group， a ZJTP drug-containing serum group， a Si-GPR43 group， and a Si-GPR43 + ZJTP drug-containing serum group. The content of IL-1β， IL-6， and TNF-α was detected by ELISA. The protein expression of pathways was detected by Western blot. IF was used to observe the nucleation of NF-κB p65. ResultThe optimal molding condition was 1 mg·L-1 LPS for 24 h. The optimal drug intervention condition was 5% ZJTP drug-containing serum for 24 h. Compared with the blank control group， the content of ET-1 in the model group was significantly increased， and the content of NO was significantly decreased （P<0.01）. The levels of inflammatory factors were significantly increased （P<0.01）. The expressions of GPR43 and IκBα were significantly decreased， while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly increased （P<0.01）. NF-κB p65 protein was transferred from the extranuclear to the intranuclear （P<0.01）. Compared with the model group， the content of ET-1 in the ZJTP drug-containing serum group was decreased， and the content of NO was increased （P<0.05）. The levels of inflammatory factors decreased （P<0.05）. The protein expressions of GPR43 and IκBα were increased， while the expressions of β-arrestin-2 and NF-κB p65 were decreased （P<0.05）. The amount of NF-κB p65 transferred from the intranuclear to the extranuclear decreased （P<0.01）. The mechanism study showed that compared with the Si-GPR43 group， the content of IL-1β， IL-6， and TNF-α were significantly decreased after treatment with ZJTP drug-containing serum （P<0.01）. The protein expressions of GPR43 and IκBα were significantly increased （P<0.01）， while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly decreased （P<0.01）. The amount of NF-κB p65 transferred from the extranuclear to the intranuclear decreased （P<0.01）. ConclusionZJTP has a protective effect on HUVECs with high glucose and LPS-induced inflammatory injury， which may be related to the regulation of GPR43/β-arrestin-2/IκBα/NF-κB pathway.

Cobalt Chloride as a Hypoxia Mimicking Agent Induced HIF-1α and mTOR Expressions of Human Umbilical Cord Mesenchymal Stem Cells

ABSTRACT Objective: To assess the effects of cobalt chloride (CoCl2) as a hypoxia mimicking agent on human umbilical cord mesenchymal stem cells (hUCMSCs) expression of HIF-1α and mTOR for use in regenerative dentistry. Material and Methods: Human umbilical cord mesenchymal stem cells were isolated and then cultured. The characteristics of stemness were screened and confirmed by flow cytometry. The experiment was conducted on hypoxia (H) and normoxia (N) groups. Each group was divided and incubated into 24-, 48-, and 72-hours observations. Hypoxic treatment was performed using 100 µM CoCl2 on 5th passage cells in a conventional incubator (37°C; 5CO2). Then, immunofluorescence of HIF-1α and mTOR was done. Data was analyzed statistically using One-way ANOVA and Tukey's HSD. Results: Significant differences were found between normoxic and hypoxic groups on HIF-1α (p=0.015) and mTOR (p=0.000) expressions. The highest HIF-1α expression was found at 48 hours in the hypoxia group, while for mTOR at 24 hours in the hypoxia group. Conclusion: Hypoxia using cobalt chloride was able to increase human umbilical cord mesenchymal stem cells expression of HIF-1α and mTOR.

Natural killer cells 56bnght16- have higher counts in the umbilical cord blood than in the adult peripheral blood

ABSTRACT Introduction and hypothesis: Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for allogeneic hematopoietic stem cell transplantation in the absence of a compatible donor. The UCB transplantation has a lower incidence of chronic graft versus host disease (GvHD), but is associated with slower engraftment and slower immune reconstitution, compared to other sources. Dendritic cells (DCs) and Natural Killer cells (NKs) play a central role in the development of GvHD and the graft versus leukemia (GvL) effect, as well as in the control of infectious complications. Method: We quantified by multiparametric flow cytometry monocytes, lymphocytes, NK cells, and DCs, including their subsets, in UCB samples from 54 healthy newborns and peripheral blood (PB) from 25 healthy adult volunteers. Results: In the UCB samples, there were higher counts of NK cells 56bright16- (median 0.024 × 109/L), compared to the PB samples (0.012 × 109/L, p < 0.0001), NK 56dim16bright (median 0.446 × 109/L vs. 0.259 × 109/L for PB samples, p = 0.001) and plasmacytoid dendritic cells (pDCs, median 0.008 × 109/L for UCB samples vs. 0.006 × 109/L for PB samples, p = 0.03). Moreover, non-classic monocyte counts were lower in UCB than in PB (median 0.024 × 109/L vs. 0.051 × 109/L, respectively, p < 0.0001). Conclusion: In conclusion, there were higher counts of NK cells and pDCs and lower counts of non-classic monocytes in UCB than in PB from healthy individuals. These findings might explain the lower incidence and severity of chronic GvHD, although maintaining the GvL effect, in UCB transplant recipients, compared to other stem cell sources.

Prevalencia de la arteria umbilical única en gestantes: su relación con otras malformaciones y el bajo peso al nacer

Introducción: La arteria umbilical única tiene una incidencia del 1 % en los recién nacidos. Se le asocia frecuentemente con gemelaridad, malformaciones y crecimiento intrauterino retardado, y constituye un factor de riesgo de prematuridad, muerte fetal y neonatal. Objetivos: Determinar la prevalencia de la arteria umbilical única en gestantes, y la asociación de esta entidad con otras malformaciones y el bajo peso al nacer. Materiales y métodos: Estudio descriptivo retrospectivo, con datos obtenidos de las historias clínicas y del modelo de seguimiento lineal existente en las consultas de genética comunitaria del municipio Matanzas, de enero de 2015 a diciembre de 2019. Resultados: La prevalencia de la arteria umbilical única fue del 0,3 %. Las malformaciones más frecuentes fueron las renales; el 27,7 % de los nacimientos fueron pretérmino y el 33,3 % de los nacidos fue con un peso inferior a 2500 g. Conclusiones: La arteria umbilical única constituye un marcador para otras malformaciones. Cuando coexisten ambas existe riesgo de prematuridad y bajo peso al nacer. Se recomienda realizar examen clínico posnatal a todo recién nacido con arteria umbilical única, pesquisando defectos renales y cardíacos.

Introduction: The single umbilical artery has an incidence of 1% in newborns. It is frequently associated with twinning, malformations and delayed intrauterine growth, and is a risk factor of prematurity, fetal and neonatal death. Objective: To determine the prevalence of single umbilical artery in pregnant women and the association of this entity with other malformations and low birth weight. Materials and methods: Retrospective descriptive study, with data obtained from medical records and the linear follow-up model existing in the community genetic clinics of the municipality of Matanzas, from January 2015 to December 2019. Results: The prevalence of the single umbilical artery was 0.3%. The most frequent malformations were renal ones; 27.7% of births were pre-term and 33.3% of those born weighed less than 2500g. Conclusions: The single umbilical artery is a marker for other malformations. When both coexist there is a risk of prematurity and low birth weight. Postnatal clinical examination is recommended for all newborns with single umbilical artery, checking for renal and heart defects.

Vitamina D y embarazo: una experiencia piloto / Vitamin D and pregnancy: a pilot experience

La insuficiencia de vitamina D (VD) en el embarazo se relaciona con una mayor incidencia de cesáreas, preeclampsia y partos prematuros. Objetivo: evaluar si el grado de insuficiencia de VD se asocia a mayor número de cesáreas y evaluar la correlación entre la 25 hidroxivitamina D (25OHD) materna y en sangre del cordón del recién nacido. Las mujeres (n=127) se dividieron según sus niveles de 25OHD (ng/mL):G1:<20 (deficiencia), G2:20-30 (insuficiencia), G3:>30 (suficiencia). Se registraron edad; edad gestacional (EG); índice de masa corporal (IMC); tensión arterial sistólica y diastólica; tipo de parto y la estación del año en que se tomó la muestra. Se determinaron calcemia (ng/mL); 25OHD; parathormona intacta (pg/mL); fosfatasa alcalina ósea (UI/L) y crosslaps (pg/mL). La edad media fue de 26±6 años y la EG de 35,8±2,7 semanas, sin diferencias entre grupos. El porcentaje de cesáreas fue mayor en G1 que en G2 y G3 (31,3%, 21,4% y 25%, respectivamente; p<0,05). El mayor porcentaje de muestras se tomó en primavera (p<0,05). No se observaron diferencias en las demás variables maternas estudiadas. La 25OHD materna correlacionó positivamente con los valores de la sangre de cordón de sus respectivos recién nacidos (r= 0,67; p<0,0001). Independientemente de la época del año y del IMC, se observó que un porcentaje significativo de las mujeres embarazadas estudiadas tenía niveles de 25OHD inferiores a 30 ng/mL. Conclusión: evidenciamos que la deficiencia de VD materna se asoció al número de cesáreas. Asimismo, los niveles séricos de 25OHD en sangre de cordón umbilical correlacionaron significativamente con los maternos. (AU)

Vitamin D (VD) insufficiency in pregnancy is associated with a higher incidence of cesarean section, preeclampsia, and preterm delivery. Objective: to evaluate if the degree of VD insufficiency is associated with the incidence of cesarean section and to determine the correlation between maternal and newborn cord blood 25-hydroxy VS (25OHD). Women (n=127) were divided according to their 25OHD levels (ng/mL): G1:<20 (deficiency), G2:20-30 (insufficiency), G3:>30 (sufficiency). Age; gestational age (GA); body mass index (BMI); systolic and diastolic blood pressure (mmHg); type of delivery and the season of the year in which the sample was taken were recorded. Calcemia (ng/mL); 25OHD; intact parathormone (pg/mL); bone alkaline phosphatase (IU/L) and Crosslaps (pg/mL) levels were determined. Mean age was 26±6 years and GA was 35.8±2.7 weeks with no differences among groups. The % of cesarean sections was higher in G1 than in G2 and G3 (31.3%, 21.4% and 25%; p<0.05). The highest % of samples were taken in spring (p<0.05). No differences were observed in the other maternal variables studied. Maternal serum 25OHD levels correlated positively with those of cord blood from their respective newborns (r=0.67; p<0.0001). Regardless the season of the year and BMI, a high % of the studied pregnant women presented 25OHD levels lower than 30 ng/ml. Conclusion: we found that maternal VD deficiency is associated with the number of cesarean sections. In addition, 25OHD levels in the newborn significantly correlate with maternal serum levels. (AU)

Lactate and pH values in newborns with a history of acute fetal distress / Valores de lactato y pH en recién nacidos con antecedente de sufrimiento fetal agudo

Abstract Background: Acute fetal distress (AFD) is a condition that requires timely diagnosis because it generates hypoxia, acidosis, and even intrauterine death. This study aimed to determine lactate and pH values in the umbilical cord in full-term newborns (NBs) with a history of AFD. Methods: We conducted a cross-sectional study in full-term NBs of mothers with at least one perinatal, neonatal, or gasometric AFD antecedent. Neonatal morbidity was considered: if 1-min Apgar ≤ 6, or advanced neonatal maneuvers, or neonatal intensive care unit (NICU) admissions were necessary. The cutoff points were lactate > 4mmol/L and pH < 7.2. Results: Of 66 NBs, 33.3% of mothers presented at least one antecedent for developing AFD; 22.7% presented hypertensive pregnancy disease, 13.6% oligohydramnios, and 63.6% other factors. Perinatally, 28.7% required advanced neonatal resuscitation maneuvers and 7.5% admission to the NICU. In the gasometry, the lactate and pH values for the neonatal morbidity of the NBs' group were 4.726 ± 1.401 and 7.293 ± 0.056, respectively, versus 2.240 ± 0.318 and 7.359 ± 0.022 (p < 0.05) for the group without associated neonatal morbidity. Conclusions: Lactate values in the umbilical cord increased by 25%, and pH decreased by one percent in NBs with a history of AFD and associated morbidity.

Resumen Introducción: El sufrimiento fetal agudo (SFA) es una condición que amerita un diagnóstico oportuno debido a que genera hipoxia, acidosis e incluso la muerte intrauterina. El objetivo de este estudio fue determinar los valores de lactato y pH en cordón umbilical en recién nacidos de término con antecedente SFA. Métodos: Se llevó a cabo un estudio transversal, en recién nacidos a término, de madres que tuvieron al menos un antecedente para SFA de tipo perinatal, neonatal o gasométrico. Se consideró morbilidad neonatal cuando presentaron Apgar al minuto ≤ 6, o requirieron maniobras avanzadas de reanimación neonatal, o ingreso a Unidad de Cuidados Intensivos Neonatales (UCIN). El punto de corte fue > 4 mmol/L para los valores de lactato y pH < 7.2. Resultados: De un total de 66 recién nacidos, el 33.3% de las madres presentaron al menos un antecedente para desarrollar SFA; el 22.7% presentó enfermedad hipertensiva del embarazo, el 13.6%, oligohidramnios, y el 63.6%, otros factores. El 28.7% requirieron maniobras avanzadas de la reanimación neonatal y el 7.5%, el ingreso a la UCIN. En la gasometría, el valor de lactato y pH para el grupo de recién nacidos con morbilidad neonatal fue de 4.726 ± 1.401 y 7.293 ± 0.056 respectivamente, versus 2.240 ± 0.318 y 7.359 ± 0.022 (p < 0.05) para el grupo sin morbilidad neonatal asociada. Conclusiones: Se observó un incremento del 25% de los valores de lactato en cordón umbilical y una disminución del 1% del pH en los recién nacidos con antecedente de SFA y morbilidad asociada.

Comparison between Wright and Shukla formulas: which is better for the placement of umbilical catheters in newborns ≤ 1500 g? / Comparación entre las fórmulas de Wright y de Shukla: ¿cuál es mejor para la colocación de catéteres umbilicales en recién nacidos ≤ 1500 g?

Abstract Background: Intravascular venous (VUC) or arterial (AUC) umbilical catheter placement is the most frequent invasive procedure in the neonatal intensive care unit (NICU). Either Wright's or Shukla's formula is used to introduce the catheters. However, Shukla's formula is associated with incorrect insertion, especially for newborns < 1500 g. This study aimed to determine by chest X-ray if Wright's formula is better than Shukla's formula for the correct placement of umbilical catheters in newborns ≤ 1500 g. Methods: We included patients admitted to the NICU of a secondary-level hospital between 2021-2022 who received VUC or AUC through the Wright or Shukla formulas. Results: A total of 129 newborns were included: 78 with VUC and 51 with AUC. In VUC, 50% with Wright and 36.8% with Shukla formulas had the correct location, (p = 0.24). In AUC, 56.6 % with Wright and 52.4% with Shukla formulas were placed correctly placed, (p = 0.76). VUC with weight < 1000 g were correctly placed in 36.4% with Wright and 33.3% with Shukla formulas (p = 0.58). VUC in newborns > 1000 g were correctly placed in 66.6% with Wright and 38.4% with Shukla formulas (p = 0.065). AUC in newborns < 1000 g were correctly placed in 45% using Wright and 42.9% Shukla formulas (p = 0.63). AUC in newborns > 1000 g were correctly placed in 80% using Wright and 57.1% Shukla formulas (p = 0.23). Conclusions: We found 13% more correctly placed VUC using Wright's formula. Moreover, Wright's formula was 29% above Shukla's VUC placement in neonates > 1000 g, although there was no significant difference due to the sample size.

Resumen Introducción: La colocación de catéteres intravasculares venosos umbilicales (CVU) y arteriales (CAU) es el procedimiento invasivo más frecuente en la unidad de cuidados intensivos neonatales (UCIN). Para introducirlos se utilizan las fórmulas de Wright y de Shukla, aunque esta última podría estar asociada con una inserción incorrecta, especialmente en neonatos < 1500 g. El objetivo de este estudio fue determinar mediante radiografía de tórax cuál fórmula es mejor para la correcta colocación de catéteres umbilicales en recién nacidos ≤ 1500 g. Métodos: Se incluyeron los pacientes ingresados en la UCIN de un hospital de segundo nivel entre 2021-2022 que recibieron CVU o CAU mediante las fórmulas de Wrigth o Shukla. Resultados: Se incluyeron en total 129 recién nacidos: 78 CVU y 51 CAU. En CVU, Wright 50% y Shukla 36.8% tuvieron localización correcta, p = 0.24. En las CAU, Wright 56.6% y Shukla 52.4% tenían una ubicación correcta, p = 0.76. En CVU con peso < 1000 g, Wright 36.4% y Shukla 33.3% bien situados, p = 0.58. En CVU > 1000 g, Wright 66.6% y Shukla 38.4% bien situados, p = 0.065. En CAU < 1000 g, Wright 45% y Shukla 42.9%, p = 0.63. En CAU con peso > 1000 g, Wright 80% y Shukla 57.1%, p = 0.23. Conclusiones: La colocación del CVU fue 13% mejor con la fórmula de Wright. La fórmula de Wright superó en el 29% la colocación del CVU en los neonatos > 1000 g en comparación con la de Shukla, aunque no hubo diferencia significativa debido al tamaño de la muestra.

Concentración de metales en sangre de cordón umbilical debido a exposición prenatal en una cohorte de la Ciudad de México / Prenatal exposure to metals and concentration thereof in umbilical cord blood in a Mexico City cohort

Resumen Antecedentes: En México es exigua la evidencia sobre la exposición prenatal a metales. Objetivo: Estimar la concentración de arsénico, cadmio, manganeso y plomo en sangre de cordón umbilical (SCU), y su asociación con las concentraciones en sangre materna durante el embarazo y parto. Material y métodos: Se analizó la concentración de los metales en sangre materna durante el embarazo (n = 901), parto (n = 732) y en la SCU (n = 512) de participantes de la cohorte PROGRESS, residentes en la Ciudad de México. Se estimó la asociación entre la concentración en SCU y los biomarcadores maternos mediante modelos lineales generalizados, ajustados por covariables relevantes. Resultados: La media (μg/L) de plomo, arsénico y manganeso en SCU fue 27.14 (25.28-29.14), 0.77 (0.71-0.84) y 42.60 (40.45-44.83), respectivamente. El valor del cadmio no se pudo estimar porque 86.2 % de las mediciones fueron inferiores al límite de detección. Las concentraciones de plomo y manganeso en SCU se asociaron significativamente a los biomarcadores maternos durante el embarazo y el parto; solo se observó asociación con arsénico en el parto. Conclusiones: La exposición prenatal a metales tóxicos en periodos sensibles de la organogénesis evidencia un problema de salud pública desatendido. Se requiere un biomonitoreo poblacional y establecer regulación dirigida a proveer atención a población vulnerable.

Abstract Background: In Mexico, there is a paucity of evidence on the magnitude of prenatal exposure to metals. Objective: To estimate the concentration of arsenic, cadmium, manganese and lead in umbilical cord blood (UCB) and its association with maternal blood concentrations during pregnancy and delivery. Material and methods: Metal concentration in maternal blood was analyzed during pregnancy (n = 901), delivery (n = 732) and in UCB (n = 512) from participants of the PROGRESS cohort residing in Mexico City. The association between concentrations in UCB and maternal biomarkers was analyzed using generalized linear models, adjusted for relevant covariates. Results: Mean concentrations (μg/L) of lead, arsenic and manganese in UCB were 27.14 (25.28-29.14), 0.77 (0.71-0.84) and 42.60 (40.45-44.83), respectively. Cadmium concentration could not be estimated because 86.2% of measurements were below the detection limit. Lead and manganese concentrations in UCB were significantly associated with maternal biomarkers during pregnancy and delivery; at delivery, association was only observed with arsenic. Conclusions: Prenatal exposure to toxic metals in sensitive periods of organogenesis shows a neglected public health problem. Biomonitoring of the population and establishment of regulations aimed at providing care to vulnerable populations is required.

ASSOCIATION OF SERUM FERRITIN IN POST COVID-19 ANAEMIC PATIENTS TREATED WITH UMBILICAL CORD BLOOD AND HAEMATINICS.

COVID-19 patients commonly present with lower respiratory symptoms with other systemic involvement. Haematological manifestation such as low haemoglobin, thrombocytopenia, lymphocytopenia also common in COVID19 patients. In this study, we investigated prevalence, association with serum ferritin in post COVID-19 anaemic patients, after human umbilical cord blood transfusion in relation to control group. Among 155 COVID-19 RT-PCR positive patients 36 (23%) was anaemic. In our study 18 patients was transfused human umbilical cord blood, 12 patients were treated with haematinics and 6 patients denied taking any of the above. In most cases anaemia was moderate to severe that may be due to inflammation or due to pre-existing iron deficiency.Umbilical cord blood transfusion to post COVID -19 patients for the treatment of anaemia because of the unique composition of UCB. Haematological analysis and serum ferritin estimation reflecting the treatment out come in post COVID-19 anaemic patients. There was a difference between the dependent variable's serum ferritin (p <.001) in anaemic COVID-19 patients. In conclusion, our result highlight serum ferritin is widely used in diagnosis and monitoring of COVID-19 disease.

Doppler Indices of the Fetal Middle Cerebral and the Umbilical Arteries in Pregnancies at High Risk Followed Up for Suspected Fetal Growth Restriction

Background: Fetal growth restriction is related to compromised perinatal outcomes. The screening and prevention tools for fetal growth restriction like Doppler indices in high-risk groups compared with general antenatal populations. An evaluation of the correlation between Doppler indices and placental weight and birth weight of the neonate at term pregnancy in high-risk pregnancies is essential. For the early detection of fetal growth limitations in high-risk pregnancies, sensitive screening techniques are few. Objectives: To determine the most accurate indicator for predicting a poor perinatal outcome or intrauterine growth restriction by comparing and correlating the modifications in Doppler ultrasound studies of fetal circulation in general pregnant women with those of high-risk patients both with and without intrauterine growth retardation. Study design: A cross-sectional research including 81 healthy pregnancies and 19 high-risk patients at 31–40 weeks of gestation was conducted. The pulsatility index (PI) of the middle cerebral artery (MCA), the umbilical artery (UA), and the MCA PI to UA PI ratio were all analyzed. We compared the Doppler indices’ mean values. Then these values were correlated with placental weight and birth weight of the offspring. Results: A significantly low birth weight and less fetoplacental ratio and placental coefficient ratio were found in high-risk cases than in normal pregnant women (P <0.05). A strong positive relationship was observed between the middle cerebral artery pulsatility index and placental weight, while negative relationship between the pulsatility index of the middle cerebral artery and the Feto-placental ratio (P < 0.05). In addition, a positive association was found between the pulsatility index of the middle cerebral artery and placental coefficient, whereas a negative correlation was observed between the Cerebro-placental ratio and Feto-placental ratio in high-risk cases (P <0.05). Conclusion: Low birth weight can be predicted using Doppler indices since there is a definite correlation between it and unfavorable perinatal outcomes.

Effects of maternal anxiety on fetal and maternal circulation

SUMMARY OBJECTIVE: The aim of this study was to evaluate the association between maternal anxiety in the third trimester and changes in fetal and maternal circulation assessed by Doppler velocimetry. METHODS: This is a prospective, cross-sectional study. The inclusion criteria were good health, a singleton pregnancy, maternal age between 18 and 40 years, and gestational age between 34 and 40 weeks. Doppler measurements included mean uterine artery pulsatility index, fetal middle cerebral artery pulsatility index, peak of systolic velocity, umbilical artery, and umbilical vein. The Beck Anxiety Inventory questionnaire, validated for the Brazilian population, with 21 self-reported items, was applied. RESULTS: The study included 34 pregnant women, and 6 (17.7%) presented a total Beck Anxiety Inventory score showing moderate or severe maternal anxiety. The mean maternal age was 28.1 years (SD 5.7 years); the mean gestational age at interview was 36.5 weeks (SD 1.8 weeks), and the mean Beck Anxiety Inventory total score was 12.3 (SD 9.8). The group with moderate or severe anxiety, compared to the group with minimal or mild anxiety, presented an association with lower maternal age (median 21.5 vs. 29.5 years, p=0.019), lower fetal umbilical vein blood flow (median 189.4 vs. 249.5 mL/min, p=0.047), and lower umbilical vein-corrected blood flow (median 68.5 vs. 84.9 mL/kg/min, p=0.038). CONCLUSION: Maternal anxiety may affect fetal circulation patterns in late pregnancy and is associated with reduced blood flow in the fetal umbilical vein. The underlying physiopathology needs further investigation.

Vasa previa con inserción velamentosa de cordón: comunicación de un caso

La afección vasa previa es un hallazgo prenatal raro y poco frecuente de hemorragia en la segunda mitad del embarazo, en la que los vasos umbilicales desprovistos de la gelatina de Wharton se interponen entre la presentación fetal y el orificio cervical interno. Cuando no se la detecta y se produce la rotura de los vasos, se asocia a una alta tasa de mortalidad perinatal. Se describen 3 tipos; el caso presentado se trata de vasa previa de tipo 1 secundaria a inserción velamentosa de cordón. Fue diagnosticada prenatalmente mediante ecografía por vía transvaginal asociada a Doppler color. Se practicó una cesárea con evolución materno perinatal favorable.

Vasa previa is a rare and infrequent prenatal finding of hemorrhage in the second half of pregnancy, in which umbilical vessels devoid of Wharton's jelly interpose between the fetal presentation and the internal cervical os. When undetected and rupture of the vessels occurs, it is associated with a high perinatal mortality rate. Three types are described; the case presented is type 1 vasa previa secondary to velamentous insertion of the cord. It was diagnosed prenatally by transvaginal ultrasound associated with color Doppler. A cesarean section was performed with favorable maternal and perinatal evolution.

Oxymatrine improves palmitic acid-induced vascular endothelial cell injury through Akt-eNOS-NO signaling pathway / 中国药理学通报

Aim To investigate the protective effect of oxymatrine (OMT) on vascular endothelial cell injury induced by palmitic acid ( PA) and its mechanism. Methods Cell viability was detected by MTT assay. Cell apoptosis was detected by flow cytometry. The levels of oxygen species ( ROS) in cells, and lactate de-hydrogenase, malondialdehyde (MDA), superoxide dismutase (SOD) , glutathione peroxidase (GSH-PX) and nitric oxide ( NO) in cell culture medium were detected by ELISA. The protein expressions of bcl-2, bax, caspase-3, Akt and eNOS in HUVECs were detected by Western blot. Results OMT significantly inhibited PA-induced decrease in cell viability and increase in level of LDH in HUVECs. OMT also significantly inhibited PA-induced increase in cell apoptosis, and up-regulated the protein expression ratio of bcl-2/ bax and down-regulated the protein expression of caspase-3. In addition, OMT reduced the levels of ROS and MDA, and increased the levels of SOD, GSH-Px and NO in cell-culture medium treated with PA. Furthermore, OMT increased the protein phospho-rylation of Akt and eNOS in injured cells. Conclusion OMT ameliorates PA-induced vascular endothelial cell injury through Akt-eNOS-NO signaling pathway.

Matrine inhibits inflammatory response induced by TNF-α in human umbilical vein endothelial cells through miR-25-3p-mediated Klf4 pathway / 中国中药杂志

This study aimed to analyze the effect of matrine on tumor necrosis factor-α(TNF-α)-induced inflammatory response in human umbilical vein endothelial cells(HUVECs) and explore whether the underlying mechanism was related to the miR-25-3p-mediated Krüppel-like factor 4(Klf4) pathway. The HUVEC cell inflammation model was induced by TNF-α stimulation. After 24 or 48 hours of incubation with different concentrations of matrine(0.625, 1.25, and 2.5 mmol·L~(-1)), CCK-8 assay was used to detect cell proliferation. After treatment with 2.5 mmol·L~(-1) matrine for 48 h, the expression of TNF-α, interleukin-6(IL-6), interleukin-1β(IL-1β), and Klf4 mRNA and miR-25-3p was detected by real-time fluorescence-based quantitative PCR, and the protein expression of TNF-α, IL-6, IL-1β, and Klf4 was detected by Western blot. The anti-miR-25-3p was transfected into HUVECs, and the effect of anti-miR-25-3p on TNF-α-induced cell proliferation and inflammatory factors was detected by the above method. The cells were further transfected with miR-25-3p and incubated with matrine to detect the changes in proliferation and expression of related inflammatory factors, miR-25-3p, and Klf4. The targeting relationship between miR-25-3p and Klf4 was verified by bioinformatics analysis and dual luciferase reporter gene assay. The results displayed that matrine could inhibit TNF-α-induced HUVEC proliferation, decrease the mRNA and protein expression of TNF-α, IL-6, and IL-1β, increase the mRNA and protein expression of Klf4, and reduce the expression of miR-25-3p. Bioinformatics analysis showed that there were specific complementary binding sites between miR-25-3p and Klf4 sequences. Dual luciferase reporter gene assay confirmed that miR-25-3p negatively regulated Klf4 expression in HUVECs by targeting. The inhibition of miR-25-3p expression can reduce TNF-α-induced cell proliferation and mRNA and protein expression of TNF-α, IL-6, and IL-1β. MiR-25-3p overexpression could reverse the effect of matrine on TNF-α-induced cell proliferation and the mRNA and protein expression of TNF-α, IL-6, IL-1β, and Klf4. This study shows that matrine inhibits the inflammatory response induced by TNF-α in HUVECs through miR-25-3p-mediated Klf4 pathway.