[Meta-analysis of Bailing Capsules in improvement of microinflammation and nutritional status among maintenance hemodialysis patients].

This Meta-analysis was designed to evaluate the effects of Bailing Capsules on microinflammation and nutritional status of maintenance hemodialysis patients, and to determine its efficacy and safety. The randomized controlled trials concerning the intervention of microinflammation and nutritional status in maintenance hemodialysis patients with Bailing Capsules were searched from Chinese and English databases including CNKI, Wanfang, VIP, PubMed, EMbase, and Cochrane Library. A total of 16 articles were obtained, involving 1 095 cases. As revealed by Meta-analysis,(1)Bailing Capsules lowered the levels of serum high sensitivity C-reactive protein(SMD=-0.92, 95%CI[-1.05,-0.80], P<0.000 01), interleukin-6(SMD=-1.49, 95%CI[-1.96,-1.02], P<0.000 01), and tumor necrosis factor-α(SMD=-1.48, 95%CI[-1.68,-1.28], P<0.000 01) in patients with maintenance hemodialysis, thus alleviating microinflammation.(2)Bailing Capsules elevated the levels of serum hemoglobin(SMD=1.37, 95%CI[1.21, 1.54], P<0.000 01), albumin(SMD=0.78, 95%CI[0.57, 0.98], P<0.000 01), and triglyceride(SMD=0.29, 95%CI[0.07, 0.50], P=0.01) in patients with hemodialysis to improve their nutritional status.(3)Bailing Capsules reduced the incidence of cardiovascular events(RR=0.45, 95%CI[0.34, 0.59], P<0.000 01).(4)A total of six patients presented with mild gastrointestinal discomfort after receiving Bailing Capsules, and no serious adverse reactions were observed. The sequential analysis showed that the sample size of this Meta-analysis had reached the expected value. Meanwhile, the grade of evidence quality suggested that the outcome indicators were mainly low or extremely low in quality. In conclusion, Bailing Capsules might have potential advantages in alleviating microinflammation, improving nutritional status, and reducing the incidence of cardiovascular events. However, in view of the low quality and evidence of the included literature, high-quality clinical trials are needed to further confirm the efficacy and safety of Bailing Capsules.

The effects of gestational diabetes mellitus with maternal age between 35 and 40 years on the metabolite profiles of plasma and urine.

BACKGROUND: Gestational diabetes mellitus (GDM) is defined as impaired glucose tolerance in pregnancy and without a history of diabetes mellitus. While there are limited metabolomic studies involving advanced maternal age in China, we aim to investigate the metabolomic profiling of plasma and urine in pregnancies complicated with GDM aged at 35-40 years at early and late gestation. METHODS: Twenty normal and 20 GDM pregnant participants (≥ 35 years old) were enlisted from the Complex Lipids in Mothers and Babies (CLIMB) study. Maternal plasma and urine collected at the first and third trimester were detected using gas chromatography-mass spectrometry (GC-MS). RESULTS: One hundred sixty-five metabolites and 192 metabolites were found in plasma and urine respectively. Urine metabolomic profiles were incapable to distinguish GDM from controls, in comparison, there were 14 and 39 significantly different plasma metabolites between the two groups in first and third trimester respectively. Especially, by integrating seven metabolites including cysteine, malonic acid, alanine, 11,14-eicosadienoic acid, stearic acid, arachidic acid, and 2-methyloctadecanoic acid using multivariant receiver operating characteristic models, we were capable of discriminating GDM from normal pregnancies with an area under curve of 0.928 at first trimester. CONCLUSION: This study explores metabolomic profiles between GDM and normal pregnancies at the age of 35-40 years longitudinally. Several compounds have the potential to be biomarkers to predict GDM with advanced maternal age. Moreover, the discordant metabolome profiles between the two groups could be useful to understand the etiology of GDM with advanced maternal age.

Plasma LncRNA MALAT1 Expressions Are Negatively Associated with Disease Severity of Postmenopausal Osteoporosis.

BACKGROUND: Long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (LncRNA MALAT1) has been proven to promote osteogenesis in different health conditions. However, the role of plasma MALAT1 in postmenopausal osteoporosis (PMOP) has not been investigated. OBJECTIVE: To investigate whether plasma MALAT1 expressions are associated with severity of PMOP. METHODS: A total of 126 patients with PMOP and 126 healthy female control individuals were drafted into study participation. Plasma MALAT1 was detected using RT-PCR. Bone formation marker bone-specific alkaline phosphatase plasma concentration was determined using chemiluminescence immunoassay. Levels of bone absorption marker cross-linked N-telopeptidases of type I collagen were measured in duplicate using enzyme immunoassay. Bone mineral density (BMD) was examined in the total hips, femoral neck, and lumbar (L1-L4) spine using dual-energy x-ray absorptiometry. We used Genant semiquantitative (GSQ) criteria to assess the degree of vertebral deformity and fracture. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the potential diagnostic value of MALAT1 with regard to the GSQ grading. We used the Visual Analog Scale (VAS) and Oswestry Disability Index (ODI) to evaluate the symptomatic severity in and functional ability of the study participants. RESULTS: Plasma MALAT1 expressions were significantly lower in patients with PMOP, compared with healthy controls. Plasma MALAT1 expressions in patients with PMOP were positively associated with total hip, femoral neck, and lumbar (L1-L4) spine BMD. In total, 95 patients experienced vertebral deformity or fracture (VF), and 31 had no fractures. Plasma MALAT1 expressions were markedly decreased in patients with VF, compared with patients without fractures. Plasma MALAT1 expressions were negatively related to GSQ grading in patients with VF. ROC curve analysis demonstrated that decreased plasma MALAT1 expression exhibits decent diagnostic value with regard to GSQ grading. Finally, we discovered that plasma MALAT1 expression was also negatively associated with VAS and ODI. CONCLUSION: Plasma MALAT1 expressions are negatively associated with severity of PMOP.

Feasibility and effectiveness of prone position ventilation technique for postoperative acute lung injury in infants with congenital heart disease: study protocol for a prospective randomized study.

BACKGROUND: Prone position ventilation is a widely used lung protection ventilation strategy. The strategy is more convenient to implement in children compared to adults. Due to the precise mechanism of improving oxygenation function, development of pediatric prone ventilation technology has been largely focused on children with acute respiratory distress syndrome. There is a paucity of high-quality studies investigating the effects of prone position ventilation after pediatric cardiac surgery. The purpose of this study is to evaluate the feasibility and effectiveness of prone position ventilation in infants who develop postoperative acute lung injury after surgery for congenital heart disease. METHODS: A single-center, randomized controlled trial of pediatric patients with acute lung injury after surgery for congenital heart disease who will receive prone position ventilation or usual care (control group). A total of 68 children will be enrolled according to the inclusion criteria. The main outcome measures will be lung compliance and oxygenation index. The secondary outcomes will be duration of mechanical ventilation, length of stay in cardiac intensive care unit, reintubation rate, and complication rate. DISCUSSION: This study will investigate the feasibility and effectiveness of prone position ventilation techniques in children who develop postoperative acute lung injury after surgery for congenital heart disease. The results may help inform strategies to improve airway management after surgery for congenital heart disease. TRIAL REGISTRATION: ClinicalTrials.gov NCT04607993 . Initially registered on 29 October 2020.

[Prevalence of peri-implantitis and peri-implant mucositis within 15 years of implant placement].

PURPOSE: To explore the incidence of peri-implantitis (PI) and peri-implant mucositis (PM) during 15 years of implant placement. METHODS: A retrospective analysis of 507 patients (1 162 implants in total) who underwent oral implant restoration in the Affiliated Stomatological Hospital of Nanchang University from January 2001 to December 2005 were performed and followed up for 12-15 years. The clinical data of the patients were collected, and the individual and implant-level PI, PM incidence and influencing factors were analyzed. SPSS 22.0 software package was used for statistical analysis. RESULTS: After an average of 13.37 years of follow-up, the overall incidence of PM and PI in 507 implant restoration patients was 45.0% and 9.7%, respectively. The incidence of PM and PI in 1 162 implants was 44.1% and 10.9%, respectively. Among 127 implants with PI, there were 8 implants (6.3%) failed. PI had a low incidence within 0.5 to 1 year after implantation and restoration, with a significant increase in incidence within 1 to 5 years, a decrease in incidence within 5 to 10 years, and a continuous decrease in incidence over 10 years. The incidence of PM was relatively high within 0.5-1 year of implantation and restoration, gradually decreased in 1-5 years, and remained basically unchanged for 5-10 years and more than 10 years. The incidence of PI and PM using Straumann system was the lowest, and the incidence of Osstem system was the highest (P<0.05). The incidence of PI and PM in the upper anterior tooth area was significantly higher than that of other teeth(P<0.05). The probability of PI and PM was the highest in patients with non-closed crown edges, followed by loose abutment screws, loose crown-retained screws, and broken abutment screws(P<0.05). Multivariate logistic regression analysis showed that implantation time, implant system, implant position, and restorative factors were high-risk factors affecting the incidence of PM and PI (P<0.05). CONCLUSIONS: The incidence of PM is widespread within 15 years of implant placement. The incidence of PI does not increase with the increase of restoration time, but is related to implantation time, implant system, implant position and later restoration factors.

A Quality Improvement Initiative to Increase Mother's Own Milk Use in a Chinese Neonatal Intensive Care Unit.

Background: To evaluate the efficacy of a multidisciplinary quality improvement intervention to promote mother's own milk feeding and reduce necrotizing enterocolitis (NEC) in very low-birth-weight infants. Materials and Methods: We conducted a pre (January 2014 to March 2015)-post (April 2015 to June 2016), nonrandomized, interventional cohort study of infants born at <1,500 g birth weight and admitted to the Fudan University Children's Hospital level III neonatal intensive care unit in Shanghai. The intervention included establishing a breast milk promotion team and breast milk pumping room, educating staff and parents, and distributing teaching materials. The primary outcome was breast milk feeding rate. Secondary outcomes included incidences of NEC, NEC needing surgery, mortality, and time to full enteral feeds. Results: A total of 488 infants (210 baseline, 278 intervention) <1,500 g were enrolled. The intervention group had significantly increased feeding rates for any mother's milk (34.76% vs. 80.58%; p < 0.01) and high-volume mother's milk (≥50% of feeds; 22.86% vs. 61.15%; p < 0.01), and decreased incidence of NEC needing surgery (7.62% vs. 3.24%; adjusted odds ratio [OR] 0.32, 95% confidence interval [CI] 0.14-0.76). There were no significant differences in rates of mortality (0.5% vs. 1.49%; adjusted OR 2.10, 95% CI 0.22-19.6), NEC (10.00% vs. 7.55%; adjusted OR 0.59, 95% CI 0.31-1.14), and time to full enteral feeds (20.18 ± 1.67 days vs. 24.15 ± 1.65 days; adjusted OR = 1.09, 95% CI 0.99-1.21). Conclusions: Our quality improvement initiative increased the consumption of mother's own milk and reduced the severity of NEC in very low-birth-weight infants.

Correlation of Serum CCL3/MIP-1α Levels with Disease Severity in Postmenopausal Osteoporotic Females

BACKGROUND: The pro-inflammatory protein chemokine cytokine ligand 3 is well established as a vital regulator of bone resorption and osteoclast stimulation. AIMS: To investigate if serum cytokine ligand 3 levels correlated with disease severity in postmenopausal osteoporotic women. STUDY DESIGN: Cross-sectional study. METHODS: Eighty-two postmenopausal osteoporotic women, 76 postmenopausal non-osteoporotic women, and 80 healthy women of childbearing age were recruited. The total hip, femoral neck, and L1-L4 spine bone mineral density were assessed by dual-energy X-ray absorptiometry. Serum cytokine ligand 3 concentrations were examined using a commercial enzyme-linked immunosorbent assay kit. Serum inflammatory cytokine interleukin-6, tumor necrosis factor-alpha, and the bone metabolic markers, carboxy-terminal crosslinked and tartrate-resistant acid phosphatase 5b were also examined. Scores on both the visual analogue scale and the Oswestry Disability Index were utilized to assess clinical severity. RESULTS: Patients in the postmenopausal osteoporotic group had significantly increased serum cytokine ligand 3 levels compared with those in both the postmenopausal non-osteoporotic group (40.9±15.1 pg/mL vs 24.2±8.7 pg/mL, p<0.001) and control group (40.9±15.1 pg/mL vs 23.9±9.1 pg/mL, p<0.001). Serum cytokine ligand 3 levels negatively correlated with bone mineral density at the total hip (r=-0.345, p=0.002), femoral neck (r=-0.329, p=0.003), and L1-L4 lumbar spine (r=-0.354, p=0.001) and positively correlated with visual analogue scale scores (r=0.413, p<0.001) and the Oswestry Disability Index (r=0.360, p<0.001). Moreover, serum cytokine ligand 3 levels were correlated with increased tumor necrosis factor-alpha (r=0.305, p=0.005), interleukin-6 (r=0.288, p=0.008), terminal crosslinked and tartrate-resistant acid phosphatase 5b (r=0.371, p<0.001), and carboxy-terminal crosslinked (r=0.317, p=0.004) levels. All correlations were still significant after adjusting for both body mass index and age. CONCLUSION: Chemokine cytokine ligand 3 may be a useful biomarker that can be used to predict disease severity of postmenopausal osteoporosis. Therapies targeting cytokine ligand 3 and its related signaling pathways to inhibit and delay the osteoclastogenesis process deserve further investigation.

Glycoengineering of pertuzumab and its impact on the pharmacokinetic/pharmacodynamic properties.

Pertuzumab is an antihuman HER2 antibody developed for HER2 positive breast cancer. Glycosylation profiles are always the important issue for antibody based therapy. Previous findings have suggested the impact of glycosylation profiles on the function of antibodies, like pharmacodynamics, antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). However, the roles of fucose and sialic acid in the function of therapeutic antibodies still need further investigation, especially the role of sialic acid in nonfucosylated antibodies. This study focused on the pharmacokinetic and pharmacodynamic properties of pertuzumab after glycoengineering. Herein, nonfucosylated pertuzumab was produced in CHOFUT8-/- cells, and desialylated pertuzumab was generated by enzymatic hydrolysis. Present data indicated that fucose was critical for ADCC activity by influencing the interaction between pertuzumab and FcγRIIIa, nevertheless removal of sialic acid increased the ADCC and CDC activity of pertuzumab. Meanwhile, regarding to sialic acid, sialidase hydrolysis directly resulted in asialoglycoprotein receptors (ASGPRs) dependent clearance in hepatic cells in vitro. The pharmacokinetic assay revealed that co-injection of asialofetuin can protect desialylated pertuzumab against ASGPRs-mediated clearance. Taken together, the present study elucidated the importance of fucose and sialic acid for pertuzumab, and also provided further understanding of the relationship of glycosylation/pharmacokinetics/pharmacodynamics of therapeutic antibody.

Establishment of a fluorescence-based method to evaluate endocytosis of desialylated glycoproteins in vitro.

Insufficient sialylation can result in rapid clearance of therapeutic glycoproteins by intracellular degradation, which is mainly mediated by asialoglycoprotein receptors (ASGPRs) on hepatic cells. In contrast, for glycoproteins, a long half-life is often related to high level of terminal sialic acid. These could be extremely important for insufficient sialylated biomedicines in clinic, and development of therapeutic glycoproteins in laboratory. However, how the desialylated glycoproteins are removed and how to evaluate the ASGPRs mediated endocytosis in vitro needs further investigate. Herein we described an integrative characterization of ASGPRs in vitro to elucidate its endocytosis properties. The endocytosis was determined by a fluorescence-based quantization method. The results showed that the ASGPRs could bind to poorly sialylated glycoproteins including asialofetuin and low sialylated recombinant Factor VIIa with a relatively higher ASGPRs binding affinity, and induce a more rapid endocytosis in vitro. Moreover, the mechanism under the internalization of ASGPRs was also investigated, which was found to depend on clathrin and caveolin. Utilizing the relative fluorescence quantification can be suitable for measurement of insufficient sialylated glycoprotein endocytosis and quality control of therapeutic glycoproteins, which could be useful for the understanding of the development of therapeutic glycoproteins.

Noninvasive Tumor Grading of Glioblastomas Before Surgery Using Arterial Spin Labeling. A Cohort Study.

OBJECTIVE: To assess the clinical value of using arterial spin labeling (ASL) technique preoperatively for non-invasive tumor grading in glioblastoma (GBM) patients. STUDY DESIGN: Forty-nine patients with GBMs were selected, including 21 patients with high-grade gliomas and 28 patients with low-grade gliomas. ASL perfusion imaging was performed with GE Signa Excite HD 3.0 T MR scanning system (GE Healthcare). Relative cerebral blood flow (rCBF) and relative tumor blood flow (rTBF) were quantified in all patients. Statistical analysis was performed with STATA version 12.0 software. Further, relevant human cohort studies published in Chinese and English languages were identified by database searches and screened. Data was extracted and meta-analysis was performed using Comprehensive Meta-analysis version 2.0 software. RESULTS: The ratios of rTBF to rCBF in the contralateral white matter, contralateral gray matter, and contralateral hemisphere of high-grade gliomas were higher than low-grade gliomas (all p < 0.05). ASL results in the solid tumors revealed that rCBF was greater in high-grade gliomas. Such differences in rCBF were not significant (p > 0.05) when 2 and 5 cm distances from tumor margin were compared. Importantly, rCBF values showed statistical differences when the 2 cm distance was compared with the 5 cm distance from tumor margin (all p < 0.05). Finally, meta-analysis results supported the conclusion that rCBF and rTBF values were significantly higher in high-grade GBM as compared to low-grade GBM. CONCLUSION: We present convincing data that ASL is highly effective in differentiating between high-grade and low-grade gliomas, and thus is a useful tool for preoperative evaluation of GBM.

[Inhibiting effect of CaMKIIN up-regulation on leukemia cells growth and its mechanism].

OBJECTIVE: To investigate the inhibitory effects of CaMKIIN on acute myeloid leukemia cell line HL-60 to explore a novel therapeutic target of leukemia. METHODS: Human CaMK II N gene expression vector pcDNA3.1/hCaMKIIN or empty vector pcDNA3.1/myc-His (-) B was transfected into HL-60 cells by Lipofectamine 2000. Human CaMK II N proteins of transfected cells were detected by Western blot. Cell proliferation affected by human CaMKIIN was determined by MTT. Colony-forming assay was performed by soft agar growth system. The cells transfected with CaMKIIN were stained with Hoechst 33342 to detect the apoptotic proportion under fluorescence microscopy. Cell cycle was analyzed by flow cytometry. RESULTS: Human CaMKIIN was stably transfected into HL-60 cells, and overexpression of human CaMKIIN inhibited the proliferation of HL-60/CaMKIIN cells compared to HL-60/mock cells and HL-60 cells [(0.44 ± 0.03) vs (0.94 ± 0.05) vs (0.94 ± 0.04), P<0.01]. The colony formation of HL-60/CaMKIIN was also markedly smaller[(21.00 ± 3.05)/500] than that of mock-transfected [(111.00±4.58)/500]] and control cells [(119.00±6.09)/500] (P<0.01). After 72 hrs-culture, the apoptotic proportion in cells transfected with CaMK II N was obviously higher than of cells transfected with mock DNA or control [(22.49 ± 2.15)% vs (7.17 ± 0.72)% vs (6.40 ± 0.55)%, P<0.01]. Up to (82.97 ± 2.90)% human CaMKIIN/HL-60 cells were arrested at G0/G1 phase, which was more than mock-transfected [(40.53 ± 2.38)%] and control cells [(41.63 ± 2.27)%] (P<0.05). Human CaMKIIN could down-regulate expression of Bcl-2 in transfected cells. CONCLUSION: CaMK IIN up-regulation could inhibit proliferation and induce apoptosis of human acute myeloid leukemia cell HL-60.

Reconstructed adeno-associated virus with the extracellular domain of murine PD-1 induces antitumor immunity.

BACKGROUND: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. MATERIAL AND METHODS: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. RESULTS: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. CONCLUSION: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

[Expression of cysteine rich 61 and vascular endothelial growth factor genes in patients with myelodysplastic syndromes and their relationship.].

OBJECTIVE: To explore the expression of Cysleine-rich 61(Cyr61) gene in the different subtypes of myelodysplastic syndromes (MDS), and the significance of Cyr61 in the genesis progression, and transformation of MDS and the relationship between Cyr61 and vascular endothelial grown factor (VEGF). METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical S-P were used to detect mRNA and protein expressions of Cyr61 and VEGF in bone marrow mononuclear cells (BMMNC) from 28 MDS, 12 acute myeloid leukemia (AML) patients, and 10 normal volunteers. RESULTS: Expressions of Cyr61 and VEGF were higher in MDS and AML patients than in controls (P < 0.05). The expressions of Cyr61 and VEGF were significantly higher in high risk group (0.3998 +/- 0.2647, 0.4775 +/- 0.1342) than that in low risk MDS group (0.2213 +/- 0.1465, 0.2872 +/- 0.2341) (P < 0.05), but no significant difference between high risk MDS and AML patients. Expressions of Cyr61 and VEGF protein were higher in MDS patients than in normal controls (P < 0.05), and were significantly higher in high risk MDS group \[(38.7 +/- 2.9)%, (43.2 +/- 2.7)%\] than in low risk group \[(31.4 +/- 3.1)%, (33.5 +/- 3.4)%\] (P < 0.05). Expressions of Cyr61 and VEGF were significantly correlated (r = 0.8762, P < 0.01). CONCLUSION: Cyr61 and VEGF may play a role in the angiogenesis and pathogenesis of MDS.

Dehydroepiandrosterone anti-atherogenesis effect is not via its conversion to estrogen.

AIM: This study was conducted to demonstrate the anti-atherosclerotic effect of dehydroepiandrosterone (DHEA) and to investigate its possible mechanisms and whether this effect is related to its conversion to estrogen. METHODS: Forty male New Zealand White rabbits aged 3 months were divided into 5 groups (n=8 per group) and fed different diets for 10 weeks. Serum lipid levels, the area of atherosclerotic lesions and the mRNA levels of monocyte chemoattractant protein-1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) in aortic lesions were measured. Then cultured vascular smooth muscle cells (VSMCs) stimulated by oxidized low density lipoprotein-cholesterol (ox-LDL) were treated by DHEA. The gene and protein expression levels of MCP-1 and VCAM-1 in VSMCs was detected. The plasmid with or without the gene of cytochrome P450 aromatase (CYP19) was transient transfected into cultured VSMCs respectively. Twenty hours later, the cells were stimulated with ox-LDL and DHEA. RESULTS: DHEA could obviously decrease the area of atherosclerotic lesions and the expressions of MCP-1 and VCAM-1 in aortic lesions. But all-trans retinoic acid (atRA) which was reported would limit restenosis after balloon angioplasty had no visible synergistic effect with DHEA. DHEA could also reduce ox-LDL-induced MCP-1 and VCAM-1 expression in untransfected or transfected VSMCs. CONCLUSION: The anti-atherosclerotic effect of DHEA had nothing to do with the catalysis of cytochrome P450 aromatase (CYP19), or was not related to its conversion to estrogen.