Fetal pleural effusion in the uterus and dyspnea after birth / 中国当代儿科杂志

Neonatal chylothorax is a common cause of neonatal congenital pleural effusion and is often caused by the accumulation of chylous fluid in the thoracic cavity due to the rupture of the thoracic duct and its branched lymphatic vessels for a variety of reasons. Neonatal chylothorax caused by malignant tumors is extremely rare, and this is the first case of neonatal mediastinal neuroblastoma with chylothorax in China. The boy was found to have pleural effusion in the left thoracic cavity in the uterus, and experienced apnea at birth, as well as dyspnea and cyanosis as the main manifestations after birth. He was diagnosed with left chylothorax based on conventional biochemical analysis of pleural effusion. After the treatment including persistent chest drainage and symptomatic and supportive treatment, the drainage of the left thoracic cavity reached a volume of 90-180 mL per day. Neonatal refractory chylothorax was considered. Chest radiograph on day 13 after birth showed lesions in the upper left lung field, and contrast-enhanced plain CT scan of the chest suggested the possibility of posterior mediastinal neuroblastoma. The autopsy confirmed giant posterior mediastinal neuroblastoma (poorly differentiated), which involved the C7-T6 spinal canal and the nearby erector spinae, with a small amount of tumor tissue in the liver and both adrenal glands. Mediastinal tumor is considered the underlying cause of chylothorax in this case.

Levels of airway inflammatory mediators in peripheral blood in infants and young children with wheezing / 中国当代儿科杂志

OBJECTIVE@#To examine the levels of airway inflammatory mediators in peripheral blood in infants and young children with wheezing and to study the possible pathogenesis of wheezing from the aspects of T helper cell 1 (Th1)/T helper cell 2 (Th2) imbalance and airway inflammation.@*METHODS@#A total of 50 children aged 1 month to 3 years with an acute wheezing episode were enrolled as the wheezing group, and 25 age-matched healthy infants were enrolled as the healthy control group. According to the number of wheezing episodes, the wheezing group was divided into a first-episode group (n=25) and a recurrent wheezing (number of episodes ≥2) group (n=25). According to the presence or absence of high-risk factors for asthma, the wheezing group was divided into a high-risk factor group (n=22) and a non-high-risk factor group (n=28). According to the results of pathogen detection, the wheezing group was divided into a positive pathogen group (n=23) and a negative pathogen group (n=27). Levels of interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13), transforming growth factor-β1 (TGF-β1), and total IgE (TIgE) in peripheral blood were measured for each group. For children with wheezing, eosinophil (EOS) count in peripheral blood was measured, and related samples were collected for respiratory pathogen detection.@*RESULTS@#The wheezing group had significantly higher levels of IL-4, IL-5, IL-13, TGF-β1, and TIgE in peripheral blood than the healthy control group (P0.05). The correlation analysis showed that in children with wheezing, EOS count was positively correlated with IL-4 level (P<0.01), IL-4 level was positively correlated with IL-5 and IL-13 levels (P<0.01), IL-5 level was positively correlated with IL-13 level (P<0.01), and IL-2 level was positively correlated with TGF-β1 level (P<0.05).@*CONCLUSIONS@#Th1/Th2 imbalance with a predominance of Th2 is observed in infants and young children with wheezing. IL-4, IL-5, IL-13, TGF-β1, and IgE are involved in the pathogenesis of wheezing in these children. Airway inflammation is also observed in these children with wheezing, but it is not associated with the number of wheezing episodes, presence or absence of high-risk factors for asthma, or results of pathogen detection.

Hyperbaric oxygen promotes the migration and differentiation of endogenous neural stem cells in neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the effects of hyperbaric oxygen (HBO) treatment on the migration and differentiation of endogenous neural stem cells (NSCs) in neonatal rats with hypoxic-ischemic brain damage (HIBD).</p><p><b>METHODS</b>Seven-day-old Sprague-Dawley rats were randomly divided into the normal control (CON), the HIBD model and the HBO groups (HBO treatment was administered at 2 ATA, once daily for 7 days within 3 hrs after HIBD). HIBD model was prepared according to the classic Rice-Vannucci method. BrdU/DCX, BrdU/beta-tubulin, BrdU/GFAP and BrdU/O4 immunofluorescence were examined by confocal microscopy in the subventricular zone (SVZ) and the cortex 7, 14 and 28 days after HBO treatment.</p><p><b>RESULTS</b>The BrdU(+)DCX(+) cells in the SVZ (84 +/- 21 cells/mm2) in the HBO group were significantly higher than those in the CON group (39 +/- 14 cells/mm2) (p<0.05) and the HIBD model group (68 +/- 17 cells/mm2) (p<0.05) 7 days after HBO treatment. Fourteen days after HBO treatment, the BrdU(+) DCX(+) cells decreased in the SVZ and more cells were observed in the cortex in the HBO group as compared with the CON group (p<0.01). The BrdU(+) beta-tubulin(+), BrdU(+)GFAP(+) and BrdU(+) O4(+) cells were observed in the cortex, and more BrdU(+)beta-tubulin(+) and BrdU(+) O4(+) cells were observed in the HBO group as compared with the CON and the HIBD model groups (p<0.05) 28 days after HBO treatment.</p><p><b>CONCLUSIONS</b>HBO treatment may promote endogenous NSCs to migrate to the cortex and differentiate into mature neurocytes in neonatal rats with HIBD.</p>

Effect of intracerebral transplantation of rat bone marrow stromal cells on brain white matter of neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of intracerebral transplantation of bone marrow stromal cells (BMSCs) on brain white matter of neonatal rats with hypoxic-ischemic brain damage (HIBD).</p><p><b>METHODS</b>Thirty-four 7-day-old neonatal rats were randomly assigned to three groups: normal control (n=10), HIBD (n=12) and HIBD+BMSCs transplantation (n=12). The HIBD and the HIBD+BMSCs transplantation group rats were subjected to left carotid artery ligation, followed by hypoxia exposure for 2 hrs, in order to induce HIBD. The rats in the HIBD+BMSCs transplantation group received transplantation of BMSCs labeled nucleus with Hochest 33324 into the left hippocampus 24 hrs after HIBD induction. Myelin basic protein (MBP) expression in the left corpus callosum and the subcortical white matter and the number of oligodendrocyte precursors positively stained O4 in the left periventricular area and the subcortical white matter were detected by immunohistochemistry at ages of 45 days.</p><p><b>RESULTS</b>The labeled BMSCs survived and were found mainly in the left hemisphere 37 days after transplantation. The positive rate of O4 expressed by the transplanted BMSCs was 3.70+/-1.09%. More hypomyelination in the left corpus callosum and the subcortical white matter, and less number of O4 positive oligodendrocytes in the left periventricular area and the subcortical white matter were found in the HIBD group compared with the normal control group (P<0.01). The HIBD rats receiving BMSCs transplantation had increased O4 positive oligodendrocytes in the left periventricular area and the subcortical white matter and improved MBP immunoreactivity in the left corpus callosum and the subcortical white matter compared with the HIBD group (P<0.01).</p><p><b>CONCLUSIONS</b>Intracerebral transplantation of BMSCs can improve brain white matter damage in neonatal rats with HIBD.</p>

Changes of Wnt-3 protein during the proliferation of endogenous neural stem cells in neonatal rats with hypoxic-ischemic brain damage after hyperbaric oxygen therapy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Previous studies suggest that hyperbaric oxygen (HBO) treatment promotes the proliferation of neurocytes in neonatal rats following hypoxic-ischemic brain damage (HIBD). The Wnt signaling pathway is associated with neurogenesis. This study examined whether HBO promoted neural stem cells (NSCs) proliferation after HIBD, and whether that the proliferation correlated with Wnt-3 protein expression.</p><p><b>METHODS</b>Seven-day-old Sprague-Dawley rats were randomly divided into three groups: normal control, hypoxia-ischemia (HI), and HI-HBO. HI was induced by the ligation of left common carotid artery, followed by a 2-hr exposure to 8% O2 in the latter two groups. HBO was administered 3 hrs after HI in the HI-HBO group for continuous 7 days (2 atmospheres absolute, once daily). The proliferating NSCs in the subventricular zone (SVZ) was examined by BrdU/nestin immunofluorescence and the expression of Wnt-3 protein in NSCs was examined by nestin/Wnt-3 immunofluorescence at 6 and 24 hrs and at 3, 7 and 14 days of HI. The cellular expressions of nestin and Wnt-3 protein were analyzed by laser scanning confocal microscopy. The linear regression analysis was used to evaluate the correlation between cellular Wnt-3 and nestin protein. The expressions of nestin and Wnt-3 protein in the ischemic cerebral hemisphere were analyzed with Western blotting.</p><p><b>RESULTS</b>The number of BrdU/nestin positive cells in the SVZ increased 3 hrs after HBO therapy, peaked at 7 days and remained at a higher level until 14 days after HBO therapy in the HI-HBO group compared with the normal control and the HIBD groups. The level of Wnt-3 protein in NSCs increased significantly 3 hrs after HBO therapy, peaked at 3 days and remained at high levels until 14 days after HBO therapy in the HI-HBO group compared with the normal control and the HIBD groups. The level of cellular nestin protein was closely correlated with the level of cellular Wnt-3 protein (r = 0.893, P < 0.05). The Western blotting analysis demonstrated increased Wnt-3 and nestin protein expressions in the ischemic cerebral hemispheres.</p><p><b>CONCLUSIONS</b>HBO treatment promotes the proliferation of NSCs in HIBD neonatal rats, which is correlated with the activation of Wnt signaling.</p>

Effect of hyperbaric oxygen therapy administered at different time on white matter damage following hypoxic-ischemic brain damage in neonatal rats / 中国当代儿科杂志

<p><b>OBJECTIVE</b>A recent study has suggested that hyperbaric oxygen (HBO) therapy administered within 3 hrs following hypoxic-ischemic brain damage (HIBD) may alleviate brain white matter damage (WMD) in neonatal rats. However it is unclear whether a delayed HBO therapy (more than 3 hrs following HIBD) has neuroprotective effects in neonatal rats. This study aimed to explore the effect of HBO therapy administered at different time points following HIBD on WMD in neonatal rats.</p><p><b>METHODS</b>The HIBD model was prepared according to the Rice-Vannucci procedure in 7-day-old Sprague-Dawley rats. HBO therapy was administered at 3, 6, 12, 24 or 72 hrs after HIBD, once daily for consecutive 7 days. T-maze test, the foot-fault test and the radial arm maze test were performed after 14 days of HIBD. Myelin basic protein (MBP) in the callositas and corpora striata was examined by immunohistochemical method 28 days after HIBD.</p><p><b>RESULTS</b>The rats receiving HBO therapy at 3, 6 and 12 hrs after HIBD performed significantly better in the T-maze test, the radial arm maze test and the foot-fault test than the untreated HIBD rats. There were no significant differences in the behavioral test results between the HBO-treated groups administered HBO at 24 and 72 hrs after HIBD and the untreated HIBD group. The MBP expression in the HBO-treated groups treated within 12 hrs after HIBD was significantly higher than that in the untreated HIBD group (P < 0.05). When the HBO therapeutic window was delayed to 24 hrs after HIBD, there were no significant differences in the MBP expression between the HBO-treated and the untreated HIBD groups.</p><p><b>CONCLUSIONS</b>HBO therapy administered within 12 hrs following HIBD can alleviate brain WMD in neonatal rats, but the efficacy of HBO therapy administered 24 hrs after HIBD does not appear to be satisfactory.</p>

The immunoregulatory function of bone marrow mesenchymal stem cells on allogeneic B lymphocytes / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the immunoregulatory function of bone marrow mesenchymal stem cells (MSCs) on allogeneic B lymphocytes in vitro, and explore its possible mechanisms.</p><p><b>METHODS</b>Human MSCs were isolated from bone marrow and expanded in vitro. The purity of MSCs were identified with the spindle fibroblastic morphology by micro-photograph and the phenotype by flow cytometry. MSCs were irradiated with 20 Gy of gamma ray to abolish proliferative capacity. The change of activated B cells proliferative capability and apoptosis with or without MSCs were examined. The effect of MSCs on activated B cells proliferation was compared between transwell cultures and non-transwell cultures. The IgG, IgA and IgM productions of B cells and the immune molecules expression with or without MSCs were assessed. RESULTS (1) MSCs could not induce the proliferation of B lymphocytes, but could suppress LPS activated B lymphocytes proliferation. (2) With the number of MSCs increased, a dose-dependent inhibitory effect was observed in B cell proliferation. MSCs could not induce B cells apoptosis. The activated B cells proliferation with MSCs in transwell culture was decreased, suggesting that MSCs inhibition of B cells might be mediated both by cell-to-cell contact and soluble factors. (3) MSCs suppressed the IgG, IgA and IgM production of B cells, but not suppressed the immune molecules HLA-DR, CD40, CD80 and CD86 expression.</p><p><b>CONCLUSION</b>Bone marrow MSCs can suppress allogeneic B lymphocytes proliferation and its secretion in vitro.</p>

Differentiation of rat bone marrow stromal cells into neural cells induced by hypoxic-ischemic brain tissue extracts in neonate rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of brain tissue extracts in neonate rats with hypoxic-ischemic brain damage (HIBD) on the differentiation of bone marrow stromal cells (BMSCs) into neural cells.@*METHODS@#Fifteen 7-day-old neonate rats were induced HIBD by left carotid artery ligation and hypoxia exposure, and another 15-day-old neonate rats were served as normal rats. The left and right brain tissue extracts of the normal and HIBD rats were prepared 24 h after the HIBD (8-day old), 72 h after the HIBD (10-day old), and 7 d after the HIBD (14-day old), respectively (n=5). The rat BMSCs of passage 3-5 were cultured in the medium with or without previous brain tissue extracts. The expressions of neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and O(4) marked oligodendrocyte were detected after 3 days by immunocytochemistry.@*RESULTS@#The expressions of NSE, GFAP and O(4) of BMSCs cultured in the medium with left or right brain tissue extracts of different day old normal rats were higher than those of BMSCs cultured without the extracts, respectively (P<0.01), and the expressions of NSE, GFAP and O(4) of BMSCs cultured in the medium with left brain tissue extracts of 8 day old and 10 day old HIBD rats were higher than those of BMSCs cultured with right brain tissue extracts of the same day HIBD rats and BMSCs cultured with left or right brain tissue extracts of the same day normal rats (P<0.01 or P<0.05). The expressions of NSE, GFAP and O(4) of BMSCs cultured in the medium with left brain tissue extracts of 8-day-old HIBD rats were higher than those of BMSCs cultured with left brain tissue extracts of 10-day-old and 14-day-old HIBD rats (P<0.01 or P<0.05).@*CONCLUSION@#The brain tissue extracts of normal and HIBD rats can induce BMSCS into neural cells, and the damaged brain tissue extracts of 8-day-old HIBD rats is the best inductor.

Congenital hyperinsulinism: a difficult and complicated case study / 中国当代儿科杂志

This paper reported a case of congenital hyperinsulinism and reviewed the relevant literatures regarding to the etiology, pathogenesis, clinical and pathological features, diagnosis and treatment of this disorder. The baby (male), with gestational age of 36 weeks and birth weight 4,200 g, was delivered by caesarean section. It presented with hypoglycemia immediately after birth (0.8 mmol/L). Through the course of the disease, the baby's blood sugar manifested with 1.2-2.8 mmol/L although glucocorticoid was administered. 10% glucose solutions were intravenously infused at a speed of 10-17 mg/(kg x min) for this patient to retain a stable blood sugar level. The plasma insulin level was 24.13 U/L and blood sugar level was 1.5 mmol/L on day 30 of his life. The ratio of plasma insulin (U/L) and plasma glucose (mg/dL) was 0.89. These results suggest an inappropriate insulin secretion resulting in persistent hypoglycemia in this baby and so it was definitely diagnosed with congenital hyperinsulinism.

Factors influencing successful isolation of mesenchymal stem cells from human umbilical cord blood / 中华儿科杂志

<p><b>OBJECTIVE</b>Emerging evidences suggest that mesenchymal stem cells (MSCs) can be isolated from human umbilical cord blood (HUCB) and cultured in vitro, the same as the MSCs derived from bone marrow. However previous attempts to isolate MSCs from UCB showed a low rate of success (less than 30%). The present study was conducted to clarify the factors that influence the yields of MSCs from HUCB of different gestational age deliveries and to observe the bioactivity of MSCs derived from UCB.</p><p><b>METHODS</b>HUCB units were divided into three groups: gestational age (GA) 40 w group (n = 11); GA 36 w group (n = 6); GA 32 w or less than 32 w group (n = 5), cultured with optimal culture conditions. The relationship of the yields of MSCs derived from HUCB with several factors such as GA, the collected volume of HUCB and the mononuclear cells (MNCs) count of UCB, and the relationship among these factors were investigated. The bioactivity was observed by drawing the growth curve, calculating the population doubling, counting the fibroblast colony forming units (CFU-F) and detecting the surface antigen expression of MSCs by flow cytometry.</p><p><b>RESULTS</b>The success rate of generating MSCs cells was up to 54.5%. There were some correlations between the success rate and such factors as the MNCs count, the GA and the volume of UCB. The rate could be enhanced to 83.3% when the MNCs count was more than 1.25 x 10(8)/L. There was a negative correlation between the MNCs count in the same HUCB volume and the gestational age. The count of CFU-F varied with gestational age, the count of CFU-F was higher in smaller gestational age than the older. In the primary culture some cells displayed a fibroblast-like morphology and expressed MSCs-related antigens CD29, CD105, and the expression rate of these antigens were enhanced from 62.1% to 85.0% in one passage. The hematopoietic cells antigens CD34 and CD45 were less than 3% all the time.</p><p><b>CONCLUSIONS</b>The success rate could be increased when the MNCs count was more then 1.25 x 10(8)/L. There was a negative correlation between the MNCs count of the same UCB volume and the gestational age, the activity to form the CFU-F of UCB varied with gestational age; isolation of MSCs from UCB of pre-term deliveries may be relatively easier as compared to those from full term deliveries.</p>

Effect of hyperbaric oxygenation on neural stem cells and myelin in neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study investigated the effect of hyperbaric oxygenation (HBO) on neural stem cells (NSCs) and myelin in neonatal rats following hypoxic-ischemic brain damage (HIBD) and aimed to explore the possible mechanism of the protective effect of HBO on HIBD.</p><p><b>METHODS</b>Seven-day-old Sprague-Dawley rat pups were randomly assigned into 4 groups: Normal control, HIBD, hyperbaric air (HBA), and HBO groups (n=30 each). The HIBD model was produced by permanent occlusion of the left common carotid artery and 2 hrs hypoxemia exposure (8% O2 at 37 degrees C). HBA and HBO treatment was administered (2 ATA, once daily for 7 days) in the HBA and HBO groups respectively 1 hr after HIBD. BrdU immunohistochemistry was used to detect the NSCs in the sub-ventricle zone (SVZ) of the lateral ventricle and the dentate gyrus (DG) of the hippocampus. The myelin damage was assessed by myelin basic protein (MBP) immunostaining.</p><p><b>RESULTS</b>The BrdU-positive cells in the SVZ and the DG of the ischemic hemisphere in the HIBD group were dramatically decreased compared with those of the Normal control group at 3 weeks post-HIBD (P < 0.01). The HBO treatment resulted in an increase of BrdU-positive cells in the DG from 153.7 +/- 37.0 to 193.7 +/- 38.8 (P < 0.05). The nestin expression in the HIBD and HBA groups was reduced compared with that in the Normal control group. There was no difference in the nestin expression between the HBO and the Normal control groups. Hypoxia-ischemia (HI) led to marked myelin damage at 1 week post-HIBD. HBO or HBA treatment alleviated the damage.</p><p><b>CONCLUSIONS</b>The HBO treatment can result in the proliferation of BrdU-positive cells and alleviate the myelin damage following HIBD in neonatal rats, thereby offering neuroprotectivity against HI insults.</p>

The effects of hyperbaric oxygen in reducing brain damage from hypoxic ischemia in neonatal rats / 中华物理医学与康复杂志

Objective To investigate the protective effects of hyperbaric oxygen(HBO)against brain dam- age from hypoxic ischemia(HIBD)in neonatal rats.Methods One hundred and seventeen 7-day-old Sprague- Dawley rats were randomly divided into 4 groups:a control group(n=32),a hypoxic ischemia brain damage group (HIBD group,n=30),a hyperbaric air group(HBA group,n=27),and a hyperbaric oxygen group(HBO group, n=28).The HIBD model was established by permanent occlusion of the left common carotid artery followed by expo- sure to a mixture of 8% oxygen/92% nitrogen for 2 h(at 37℃).HBO therapy was administered to the HBO group after the hypoxia exposure once a day for 7 d,as was HBA therapy to the HBA group.Apoptotic cells in the cortex and hippocampus(A_(CH)cells)were measured using TUNEL at 9 d after birth,and the ratios of left and right cerebral hemisphere weight(R_(L/R))and rate of weight gain(GRW)were recorded 14 d after birth.A radial arm maze acquisi- tion test(RAMAT)was administered at 30 to 35 days.Lastly,the neuron density in the CA_1 subfield of the rats' hip- pocampi(ND_(CAI)was measured with Nissl staining.Results R_(L/R)and GRW in the HIBD group were significantly lower than in the control group(P＜0.01),while R_(L/R)was increased in the HBO and HBA groups,especially in the HBO group(P＜0.01),although there was no significant difference in GRW between the groups.Compared with the control group,A_(CH)cells were increased and ND_(CAI)was decreased in the HIBD group(P＜0.01),while A_(CH)cells were decreased and ND_(CAI)was elevated in the HBO group in comparison with the HIBD group(P＜0.01).There was no change in A_(CH)cells or ND_(CAI)in the HBA group.The RAMAT results for the HIBD group,including the time to find the arms baited with water,average times of working errors and reference memory errors,were significantly high- er than those of the control group,while these values for the HBO group were obviously lower than for the HIBD group,and there was no change for the HBA group(P＞0.05).Conclusion HBO therapy might increase the re- covery of learning and memory function by attenuating HIBD in neonatal rats.

Protective effect and mechanism of pretreatment with curcumin on infectious brain edema in rats / 中华儿科杂志

<p><b>UNLABELLED</b>Curcumin is a natural compound extracted from the spice tumeric, possessing both anti-inflammatory antioxidant, and anti-carcinogenic effect, is a potent stimulator of the stress-induced expression of heat shock protein 70 kd (HSP70).</p><p><b>OBJECTIVES</b>To study the protective effect of pretreatment with curcumin against infectious brain edema in rats, and investigate its mechanism by assessing the free radical, cytokine and HSP70 expression of the brain.</p><p><b>METHODS</b>An animal model of infectious brain edema induced by injecting pertussis bacilli (PB) through carotid artery was used. SD rats were randomly divided into five groups: (1) Normal control group (NS group, n = 9); (2) Infectious brain edema group (PB group, n = 12); (3) DMSO control group (DMSO group, n = 9); (4) HS pretreatment group (HS group, n = 9); (5) Curcumin pretreatment group (CUR group, n = 13). The water content (WC), Na(+) and K(+) content in brain tissue were measured. The content of malondialdehyde (MDA) and super oxide dismitase (SOD) were assessed by chemical colorimetry. The levels of tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were detected by ELISA. The HSP70 expression was examined by Western blot analysis.</p><p><b>RESULTS</b>(1) The WC and Na(+), MDA, TNF-alpha and IL-1beta were increased in PB group compared with NS group (P < 0.01); they were decreased in HS and CUR groups compared with PB group (P < 0.01 or P < 0.05). (2) The content of SOD was decreased in PB group than in NS group (P < 0.05), and was increased in HS and CUR group Compared with PB group, (P < 0.05). (3) Western blot analysis showed that the band density areas of HS, CUR and PB groups were higher than those in NS and DMSO groups, especially in CUR group (P < 0.01).</p><p><b>CONCLUSION</b>Pretreatment with curcumin showed a protective effect against infectious brain edema in rats. The effect might be associated with antioxidant, inhibition of the activity of cytokines and inducing expression of HSP70 by curcumin.</p>