BACKGROUND:Establishment of in vitro culture system of human placental microvascular endothelial cel s with high purity is very important. In recent studies, some scholars have successful y obtained a large number of placental microvascular endothelial cel s by three-stepenzyme digestion and magnetic separation method, but the procedures were extremely complex and it had great damage to the cel s. Therefore, how to separate human placental microvascular endothelial cel s easily and obtain high-purified cel s has become a research hotspot.
OBJECTIVE:To investigate an efficient method to isolate and purify human placental microvascular endothelial cel s from early vil us microvessels, observe the cel growth and identify the cel s.
METHODS:The vil i from normal early pregnancies (6-8 weeks) after artificial abortion were col ected aseptical y. Using two-step digestion procedure and discontinuous Percol density gradient centrifugation method, human placental microvascular endothelial cel s were obtained. Then the cel s were identified by trypsin digestion method and repeated adherence method.
RESULTS AND CONCLUSION:Human placental microvascular endothelial cel s were isolated successful y from early vil i. The primary cel s adhered to the wal s after inoculated for 24 hours and entered logarithmic phase at 10 days. 80%of the cel s achieved a confluence at 12-13 days after inoculating. The subculture cel s grew swiftly with the typical cobblestone appearance. Immunofluorescence staining showed that, cultured human placental microvascular endothelial cel s demonstrated a strong positive reaction to von Wil ebrand factor antigen and CD31, accounting for 100%. MTT assay results showed that, human placental microvascular endothelial cel s at passage 5 exhibited an S-shaped growth curve. High-purity human placental microvascular endothelial cel s can be obtained by proteolytic enzymes digestion and discontinuous Percol density gradient centrifugation method, and the purity is detected by trypsin digestion method and repeated adherence method.

Objective:To explore the prognosis and treatment experience of fetal/neonatal ovarian cyst.Methods:Clinical data of 35 cases of fetal/neonatal ovarian cyst (38 ovarian cysts) admitted to Guangdong Women and Children Hospital from June 2014 to December 2019 were retrospectively collected, including the cyst size before and after birth, ultrasonic features, intraoperative conditions, and pathology. According to the ultrasonic features at the first prenatal detection, the ovarian cysts were divided into two groups: simple cyst group (25 cysts) and complex cyst group (13 cysts). Two independent samples t-test and Fisher exact test were used to compare the characteristics of cysts between the two groups. The outcomes and treatment experience were summarized. Results:(1) The ratio of intraoperative torsion in the complex cysts group was higher than that in the simple cysts group [10/13 vs 32% (8/25), Fisher exact test, P<0.05]. (2) Twenty-five simple cysts were found on the first prenatal ultrasound scan, and 32% (8/25) of them eventually transformed into complex cysts. Among these eight cysts, the maximum diameter of five cysts was >4 cm before the transformation. (3) Postnatal ultrasound found one cyst regressed spontaneously and among the remaining 37 cysts, simple and complex type cysts were accounted for 16 and 21, respectively. Among the complex type cysts, 90% (19/21) were consistent with prenatal ultrasound. (4) Out of the 21 complicated cysts, 19 were surgically removed; the remaining two cysts (maximum diameter <3 cm) were observed conservatively and disappeared spontaneously within one year. During the operation, 81% (17/21) of the complicated cysts were found with torsion and 24% (5/21) with ovarian loss. Conclusions:Simple cysts can transform into complex cysts, especially the biggest diameter >4 cm. Complex fetal/neonatal ovarian cysts indicated by ultrasonography were more prone to torsion, which required postnatal operation.

Objective:To summarize the perinatal management and prognosis of congenital chylous ascites (CCA).Methods:Clinical data of 20 infants diagnosed with CCA and treated in Guangdong Women and Children Hospital from June 2015 to November 2020 were retrospectively analyzed and described.Results:There were ten patients with isolated CCA and ten with non-isolated CCA. In isolated CCA cases, seven were cured after conservative treatment and three after postoperative conservative treatment following an ineffective surgery. Non-isolated CCA cases were complicated by intrauterine cytomegalovirus infection ( n=1), malrotation of intestine ( n=4) or bilateral chylothorax ( n=5). In addition to conservative treatment for CCA, non-isolated CCA patients also received antiviral therapy, Ladd's procedure or intrauterine/extrauterine closed thoracic drainage. Of eight patients who were firstly diagnosed with ascites before 30 gestational weeks, including four isolated and four non-isolated cases, only one underwent surgical intervention. During hospitalization, serious infections occurred in three infants with isolated CCA and two with non-isolated CCA, and were all controlled by anti-infection treatment. During a follow-up to median age of 29 months (15-82 months), none of the patients had any abnormalities except for the one with intrauterine cytomegalovirus infection who was deaf at the age of two. Conclusions:Conservative management is effective and the prognosis is generally good for most cases with isolated CCA. Treatment and prognosis of non-isolated CCA depend on its comorbidities. Gestational age at diagnosis may not be a prognostic predictor.