The impact of excessive gestational weight gain on fetal hepatic lipid metabolism and the regulatory mechanism / 中华实用儿科临床杂志

Objective:A rat model of excessive gestational weight gain (EGWG) was constructed to investigate the impact of EGWG on fetal hepatic lipid metabolism and the relevant regulatory mechanism.Methods:Healthy Sprague-Dawley rats were caged together and tested for pregnancy.Rats with the sperm observed under microscope were considered pregnant for 0.5 days.Pregnant rats were divided into the normal diet (ND) group and high-fat diet (HFD) group by the random number table method, with 8 rats in each group.The body weight during pregnancy of the pregnant rats was recorded.Cesarean section was performed at day 21.5 of gestation and the birth weight of the fetal rats was recorded.Hepatic lipid deposition of the pregnant and fetal rats was examined by hematoxylin-eosin (HE) staining and oil red O staining.Triglyceride (TG) and cholesterol (TC) levels in livers and serum of the pregnant and fetal rats were detected by glycerol phosphate oxidase-peroxidase(GPO-PAP) method.The mRNA and protein expression levels of key genes FASN and SREBP1c in hepatic lipid metabolism of fetal rats were measured by real-time polyme-rase chain reaction (RT-PCR) and Western blot.Differences between the two groups were compared by independent sample t test. Results:There was no difference in pre-pregnancy body weight between the HFD group and the ND group, but the differences in the weight and the weight gain during pregnancy gradually enlarged between the two groups.At day 21.5 of gestation, the weight of the pregnant rats[(467.75±22.05) g vs.(430.88±18.80) g, t=-3.600, P=0.003], the weight gain of the pregnant rats during pregnancy[(181.50±9.68) g vs.(148.50±10.86) g, t=-6.415, P<0.001] and the birth weight of the fetal rats[(5.51±0.17) g vs.(4.85±0.35) g, t=-4.779, P<0.001] of the HFD group were significantly higher than those of the ND group.Both HE staining and oil red O staining presented increased hepatic lipid deposition in the pregnant and fetal rats of the HFD group.The hepatic and serum TG and TC levels of the pregnant and fetal rats of the HFD group were significantly higher than those of the ND group (all P<0.05). RT-PCR and Western blot showed that the mRNA and protein levels of key genes FASN and SREBP1c in hepatic lipid metabolism of fetal rats of the HFD group were significantly higher than those of the ND group (all P<0.05). Conclusions:An EGWG model can be successfully constructed by a 21-day HFD during pregnancy.EGWG can lead to hepatic lipid deposition in the fetal rats.The mechanism may be related to the expression changes of key genes FASN and SREBP1c in hepatic lipid metabolism of fetal rats.

Effects of p38MAPK inhibitor on fetal lung injury in a rat model of acute pancreatitis in late pregnancy / 中华急诊医学杂志

Objective@#To investigate the effect and underlying mechanisms of p38 mitogen-activated protein kinase inhibitor SB203580 on fetal lung injury in a rat model of acute pancreatitis in late pregnancy.@*Methods@#Twenty-four pregnant Sprague-Dawley rats in last gestation were randomly(random number) divided into the SO group, APILP group, and SB203580 treatment (SB) group. APILP model was induced by retrograde injection of 5% sodium taurocholate into the biliary-pancreatic duct. SB203580 administration (10 mg/kg body weight, intraperitoneal injection) was performed 0.5 h before surgery. All the rats in the SO and APILP groups received intraperitoneal injection of equivoluminal solvent at the same time point. Animals were sacrificed at 12 h after the induction of APILP, then the blood and tissue samples were harvested. Serum levels of AMY and TNF-α were analyzed. Histopathological changes of maternal pancreas and fetal lung were observed and evaluated. The expression and location of NF-κB in fetal lungs were detected by immunohistochemistry and MPO expression in fetal lungs was examined by immunofluorescence. The expression of p-p38MAPK, p38MAPK, TNF-α and ICAM-1 was determined by Western blot. One-way ANOVA and Tukey's multiple comparison tests were used for statistical analysis.@*Results@#The levels of AMY and TNF-α in maternal serum were markedly increased after APILP [(7 871.3±623.5) vs (1 915.3±452.3), (193.8±25.4) vs (107.0±13.3), (P<0.05)]. Obvious pathological changes presented in maternal pancreas and fetal lung after the attack of APILP, and their pathological scores were significantly higher than those of the SO group [(12.44±1.08) vs (1.56±0.56), (2.50±0.53) vs (0.88±0.64), (P<0.05)]. The number of NF-κB and MPO positive cells in fetal lungs were significantly higher than those in the SO group [(150.63±34.58) vs(29.50±8.80), (53.38±8.30) vs (11.75±3.33); P<0.05)]. In addition, the expression and nuclear translocation were pervasive in fetal lungs in the APILP group. Furthermore, the levels of p-p38MAPK [(0.6367±0.0386) vs (0.2282±0.0220)], TNF-α [(0.6313±0.0395) vs (0.0725±0.0076)], ICAM-1 [(0.8958±0.0776) vs (0.1372±0.0388)] and HMGB1 [(0.6478±0.0209) vs (0.2825±0.0533)] expression in fetal lungs were significantly increased after the establishment of APILP model (P<0.05). However, with the pre-administration of SB203580, the pathological scores of maternal pancreases (9.38±1.58) and fetal lungs (1.63±0.52) were decreased significantly (P<0.05), as well as the levels of AMY (4162.1±642.1) and TNF-α (139.6±21.1) in maternal serum (P<0.05). The number of NF-κB (93.00±18.88) and MPO (27.38±4.75) positive cells in fetal lungs were dramatically reduced (P<0.05) and fewer nuclear translocation was observed in the SB group. Interestingly, the expression levels of p-p38MAPK (0.2578±0.0170), TNF-α (0.3240±0.0326), ICAM-1 (0.4177±0.0823) and HMGB1 (0.4923±0.0457) in fetal lungs were markedly decreased with the treatment of SB203580 (P<0.05).@*Conclusions@#P38MAPK and its downstream inflammatory signaling pathway were involved in the process of APILP-related fetal lung injury; SB203580 administration could significantly attenuate fetal lung injury induced by APILP, which may be closely related to the inhibition of p38MAPK phosphorylation and inflammatory cascade caused by the activation of downstream signal pathways.

Effects of p38MAPK inhibitor on fetal lung injury in a rat model of acute pancreatitis in late pregnancy / 中华急诊医学杂志

Objective To investigate the effect and underlying mechanisms of p38 mitogenactivated protein kinase inhibitor SB203580 on fetal lung injury in a rat model of acute pancreatitis in late pregnancy.Methods Twenty-four pregnant Sprague-Dawley rats in last gestation were randomly(random number) divided into the SO group,APILP group,and SB203580 treatment (SB) group.APILP model was induced by retrograde injection of 5％ sodium taurocholate into the biliary-pancreatic duct.SB203580 administration (10 mg/kg body weight,intraperitoneal injection) was performed 0.5 h before surgery.All the rats in the SO and APILP groups received intraperitoneal injection of equivoluminal solvent at the same time point.Animals were sacrificed at 12 h after the induction of APILP,then the blood and tissue samples were harvested.Serum levels of AMY and TNF-α were analyzed.Histopathological changes of maternal pancreas and fetal lung were observed and evaluated.The expression and location of NF-κB in fetal lungs were detected by immunohistochemistry and MPO expression in fetal lungs was examined by immunofluorescence.The expression ofp-p38MAPK,p38MAPK,TNF-α and ICAM-1 was determined by Western blot.One-way ANOVA and Tukey's multiple comparison tests were used for statistical analysis.Results The levels of AMY and TNF-α in maternal serum were markedly increased after APILP [(7871.3±623.5) vs (1 915.3±452.3),(193.8±25.4) vs (107.0±±13.3),(P＜0.05)].Obvious pathological changes presented in matemal pancreas and fetal lung after the attack of APILP,and their pathological scores were significantly higher than those of the SO group [(12.44±1.08) vs (1.56±0.56),(2.50±0.53) vs (0.88±0.64),(P＜0.05)].The number of NF-κB and MPO positive cells in fetal lungs were significantly higher than those in the SO group [(150.63±34.58) vs(29.50±8.80),(53.38±8.30) vs (11.75±3.33);P＜0.05)].In addition,the expression and nuclear translocation were pervasive in fetal lungs in the APILP group.Furthermore,the levels of p-p38MAPK [(0.6367±0.0386) vs (0.2282±0.0220)],TNF-α [(0.6313±0.0395) vs (0.0725±0.0076)],ICAM-1 [(0.8958±0.0776) vs (0.1372±0.0388)] and HMGB1 [(0.6478±0.0209) vs (0.2825±0.0533)] expression in fetal lungs were significantly increased after the establishment of APILP model (P＜0.05).However,with the pre-administration of SB203580,the pathological scores of matemal pancreases (9.38±1.58) and fetal lungs (1.63±0.52) were decreased significantly (P＜0.05),as well as the levels of AMY (4162.1±642.1) and TNF-α (139.6±21.1) in maternal serum (P＜0.05).The number of NF-κB (93.00±18.88) and MPO (27.38±4.75) positive cells in fetal lungs were dramatically reduced (P＜0.05) and fewer nuclear translocation was observed in the SB group.Interestingly,the expression levels of p-p38MAPK (0.2578±0.0170),TNF-α (0.3240±0.0326),ICAM-1 (0.4177±0.0823) and HMGB1 (0.4923±0.0457) in fetal lungs were markedly decreased with the treatment of SB203580 (P＜0.05).Conclusions P38MAPK and its downstream inflammatory signaling pathway were involved in the process of APILP-related fetal lung injury;SB203580 administration could significantly attenuate fetal lung injury induced by APILP,which may be closely related to the inhibition of p38MAPK phosphorylation and inflammatory cascade caused by the activation of downstream signal pathways.

Neuroprotective effect of propofol on fetal ratbrain in intrauterine ischemia/reperfusion injury / 中国药理学通报

Aim To observe the neuroprotective effect of different doses of propofol on ischemic fetal rat brain.Methods Eighteen healthy pregnant SD rats were randomly allocated into the following six groups with three rats in each.Group S: sham operation group, Group IR: ischemia/reperfusion group, Group P1~P3: different doses of propofol groups, Group B: bicuculline group.In group S and group IR, 1 ml saline solution was administered via caudal vein.In group P1~P3, 10, 30, 50 mg·kg-1 of propofol was administered via caudal vein respectively.In group B, when 50 mg·kg-1 propfol was administered via caudal vein, 5 mg·kg-1 bicuculline was injected intraperitoneally at the same time.Bilateral uterine ovarian arteries were clamped for 11 mins to make intrauterine distress model of the fetal rats.The brains of fetal rats were removed after 3 days of reperfusion.Brain sections(5 μm thick) were mounted and stained with Hematoxylin and eosin(HE).The profile of the hippocampus CA1 was evaluated under a light microscope and neuronal Lesion-index(LI) was calculated.MDA content of fetal rat brain was detected by thiobarbituric acid reaction method to determine the lipid peroxidation degree of brain.Results LI was (7.2±0.9) and MDA was (3.86±0.20) μmol·g-1 in group S.LI was 71.9±2.8 and the content of MDA was (9.10±0.45) μmol·g-1 in group IR, which increased significantly compared with those in group S(P<0.01).LI was (40.8±2.6), (21.4±1.4), (20.1±1.3) and the content of MDA was (7.32±0.41), (5.65±0.27), (5.44±0.28) μmol·g-1 in propofol groups, which decreased significantly compared with those in group IR(P<0.05).LI and the content of MDA was (51.2±2.3), (7.54±0.31) μmol·g-1 in group B,respectively, reversing partly the neuroprotevtive effect of propofl.Conclusion Propofol could protect the neurons in hippocampus CA1 region of fetal rat against intrauterine distress by reducing the concentration of MDA in the brain.

Penehyclidine hydrochloride induces cerebral ischemia-reperfusion injury of fetal rats through inhibition of NF-κB signaling pathway / 临床麻醉学杂志

Objective To investigate the effects of penehyclidine hydrochloride (PHCD)on cerebral ischemia-reperfusion injury induced by intrauterine distress in fetal rats.Methods Eighty mature fetal rats weighing 4.52-4.81 g were randomly divided into four groups (n =20):sham opera-tion group(group S),PHCD control group (group S+ P),cerebral IR group (group IR),PHCD treatment group(group IR+P).Fetal rat intrauterine distress model was set up by clamping bilateral uterine horn vessels of pregnant rats.PHCD 2 mg/kg was injected in pregnant rat’s gluteus at 30 min before intrauterine distress model was set up in group IR+P,the same volume saline was injected in pregnant rat’s gluteus before shame operation in group S,the same volume PHCD was injected in pregnant rat’s gluteus before shame operation in group S+P.Fetal rats were decapitated at 12 h after the reperfusion,the peripheral blood of fetal rats was detected by blood gas analysis (including PH, PaO 2 ,PaCO 2 ,Lac);the infarct volume and the infarct volume fraction were detected by TTC stai-ning;pathological changes in lung tissue were observed by HE staining;the TNF-α,IL-6 content in the brain were detected by ELISA;the expression of NF-κB mRNA was detected by quantitative Real-time PCR,the expression of NF-κB p65 protein was detected by Western-blotting.Results The blood PH,PaO 2 in group IR and IR+P were lower than group S and S+P,the blood PH,PaO 2 in group IR+P was higher than group IR.Compared with group S and group S+P,the blood PaCO 2 , Lac,the infarct volume and the infarct volume fraction,the concentration of TNF-αand IL-6,the ex-pression of NF-κB mRNA and protein were significantly increased in group IR and IR+P (P <0.05), and those in group IR+P were lower than group IR (P <0.05 ).The pathological changes in brain tissue were significantly attenuated in group IR + P (P < 0.05 ).Conclusion Pretreatment with PHCDcouldattenuatecerebralischemia-reperfusioninjuryoffetalratsinducedbyintrauterinedistress. ThemechanismscouldrelatetotheinhibitionofNF-κBsignalingpathwayinbraintissues.

Effects of prenatal taurine supplementation on the expression of protein kinase A-Ca2 +/calmodulin-dependent protein kinase Ⅱ, c-fos in fetal rat brain with intrauterine growth restriction and its significance / 中华实用儿科临床杂志

Objective To explore the effects of prenatal taurine supplementation on the expression of protein kinase A(PKA) and c-fos,Ca2+/calmodulin-dependent protein kinase Ⅱ (GaMK Ⅱ) in fetal rat brains with intrauterine growth restriction and its significance.Methods Fifteen pregnant Sprague-Dawley rats were randomly divided into 3 groups:the control group,the intrauterine growth restriction (IUGR) group and the IUGR with prenatal taurine supplementation group(the taurine group),with 5 fetal rats in each group.All the fifteen fetal rat brains were detected as following:the expression of PKA,c-fos,CaMK Ⅱ mRNA in fetal rat brains was detected by way of real-time polymerase chain reaction(real-time PCR),while the expression changes of PKA,CaMK Ⅱ,and c-fos protein in fetal rat brains were detected by using Western blot,and the number of PKA,CaMK Ⅱ,c-fos positive cells in fetal rat brains was detected by using immunohistochemistry.Results The control group,the IUGR group and the taurine group:Comparison of the expression of PKA,CaMK Ⅱ,c-fos mRNA among 3 groups were of significant differences(F =7.934,P =0.021 ; F =5.568,P =0.043 ;F =7.332,P =0.024).Comparison of the expression of PKA,CaMK Ⅱ,c-fos protein among the 3 groups were of significant differences(F =57.743,P =0.000 ; F =163.405,P =0.000 ; F =160.136,P =0.000).Comparison of the number of PKA,CaMK Ⅱ,c-fos positive cells among the 3 groups were of significant differences (F =42.903,P =0.000 ;F =329.123,P =0.000 ; F =43.674,P =0.000).Compared with the control group,the expression of mRNA,protein,positive cells of three indicators in IUGR group was less,and the difference was statistically significant (P ＜ 0.05).The expression of mRNA,protein,positive cells of three indicators in fetal rat brains of the taurine group were not different from the control group.The expression of mRNA,protein,positive cells of three indicators in fetal rat brains of the taurine group were significantly more than the IUGR group(P ＜ 0.05).Conclusions Prenatal taurine supplementation can improve PKA-CaMK Ⅱ,c-fos mRNA and protein levels in fetal rat brain tissue with IUGR,and increase its number of positive cells and may enhance the regenerative capacity of the central nervous system,so as to reduce the IUGR brain injury and promote its role in brain development.

Effect of antenatal taurine supplementation on the taurine levels in brain of fetal rat with intrauterine growth restriction / 中华实用儿科临床杂志

Objective To examine the taurine levels and determine the influence of antenatal taurine on the taurine content in the brains of fetal rats with intrauterine growth restriction (IUGR).Methods Fifteen pregnant rats were randomly divided into 3 groups:normal control group,IUGR group and the IUGR + antenatal taurine supplement group(taurine group) (n =5).IUGR models were induced by low protein diet throughout gestation period.Taurine was added to the diet of the taurine group with a dose of 300 mg/(kg · d) from 12 days after conception until natural delivery.Two fetal rats were randomly selected from each nest and were sacrificed to obtain the brains,and the taurine levels in fetal rat brains were detected by high performance liquid chromatography-mass spectrometry.Results The average weight of fetal rats in the normal control group,the IUGR group and the taurine group were (6.619 ± 0.413) g,(4.509±0.454) g,(5.176 ±0.436) g,there was a significant difference among the 3 groups(F =429.818,P ＜ 0.01).The taurine levels in fetal rat brain in the normal control group,the IUGR group,and the taurine group were (2.399 ±0.134) × 103 μg/g,(1.881 ±0.166) × 103 μg/g and(2.170 ±0.191) × 103 μg/g,there was a significant difference among the 3 groups(F =24.828,P ＜ 0.01).Conclusion Antenatal taurine supplementation can significantly increase the taurine level in fetal rat brain with IUGR.

Isolation and primary culture of typeⅡ alveolar epithelial cells from fetal rat lung / 重庆医科大学学报

Objective:Todevelop the isolation and purification technology for typeⅡalveolar epithelial cells(AECⅡ)of fetal rat,in order to provide experimental means for the study of lung development or neonatal lung disease.Methods:Lung tissues of 19-day fetal rats were digested with trypsin and collagenase,then purified for AECⅡwith different centrifugal force and repeated attachment.Cell viability was evaluated by typran inclusion dying before plated into six-well flask.Growth status and shape of attached cells were observed with inverted phase contrast microscope.AECⅡwere identified by electron microscopy and its percentage was assessed by modified Papanicolaou dying and immunofluorescence,the latter aiming to detect expression of surfactant protein C(SPC)in AECⅡ.Results:The total amount of cells we harvested from 3 to5 fetal rat was(36?5)?106 with a viability of 98%?2%.Cells were like polygonal and connected like island under microscope.AECⅡwas ascertained by lamellar bodies found in cytoplasma under electron microscope and its percentage was 96%?3%identified by modified Papanicolaou dying,the same as the result by immunofluorescence for SPC.Conclusion:Highly Purified and viable AECⅡcould be achieved by our methods and the cell model could be used in further study.

The effect of antenatal administration with dexamethasone on the concentrations of vitamin A in fetal rats / 重庆医科大学学报

Objective:To study the effect of antenatal administration with Dexamethasone (Dex) on the vitamin A concentrations of tissues in fetal rats.Methods:Twelve pregnant Sprague-Dawley (SD) rats were randomly divided into 2 groups,one was Dex group,the other was control group.From the gestational day 16 to 18,the rats of the two groups were given Dex and Saline by intramuscular injectionrespectively.On gestational day 19,fetal rats were obtained by cesarean section,then the vitamin A concentrations of tissues (lung,liver) were detected by high-performance liquid chromatography(HPLC).Results:(1)Compared with the two groups,the per unit vitamin A concentrations of lung and liver in Dex group were higher than those of the control groop(P

The effect of NOS inhibitors on fetal rat neuron apoptosis during intrauterine distress and childhood's learning and memory / 重庆医科大学学报

Objective:Evaluating the effect of nitric oxide synthase(NOS) inhibitors on fetal rat neuron apoptosis during intrauterine distress and childhood's learning and memory.Methods:① Set up fetal rat intrauterine distress model.②Divide into the control group,reperfusion group,treatment group [injected N omega-nitro-L-arginine(L-NNA) 4mg/kg into pregnant rat's abdomen before ischemia and injected aminoguanidine(AG) 500mg/kg before operation],long-term observation group Ⅰ?Ⅱ?Ⅲ(the part of rats in control group,reperfusion group,treatment group were performed cesarean section on pregnant 19 days and bred to 40 day of age after delivery).③Measuring the content of cytochrome c in cytosol and examined the ability of learning and memory function.Results:①The content of cytochrome c in reperfusion group(6/12/24h) and treatment group(6/12/24h) were higher than those in the control group(P0.05).(3) Crossing platform times in long-term observation groupⅡ were fewer than those in long-term observation group Ⅰ?Ⅲ(P 0.05).Conclusion:NOS inhibitors play a obvious protective role in long-term intelligence of fetal rat after delivery through relieved neuron apoptosis during hypoxic ischemia encephalopathy.NOS inhibitors can be a valuable therapy method in intrauterine distress treatment.

Changes of the expressions of endoplasmic reticulum chaperones GRP78 and caspase-9 and-12 in fetal rats with intrauterine distress / 医学研究生学报

Objective:To investigate the role of glucose-regulated protein 78(GRP78) and caspase-9 and-12 in intrauterine distress-induced hypoxic-ischemic cerebral damage in fetal rats.Methods:The fetal rat model of intrauterine distress was constructed and the fetal rats were randomly divided into a normal,a sham operation and an ischemia-reperfusion(IR) group.Neuron apoptosis was analyzed by in situ end-labeled DNA(TUNEL).The expressions of caspase-9 and-12 and GRP78 proteins in the hippocampus CA1 area were detected by immunohistochemical staining.Results:Ischemia-reperfusion damage induced classic neuron apoptosis and the number of the apoptotic neurons in the hippocampus CA1 area increased with the progression of reperfusion.The expressions of GRP78 and caspase-9 and-12 were weak in the normal and sham operation group.In the IR group,the expression of GRP78 reached the peak value 3 hours after the reperfusion and then decreased gradually;the intensity of caspase-12 was increased rapidly while that of caspase-9 elevated very little within 3 hours,but both reached the peak value at 12 hours.Conclusion:Intrauterine distress-induced hypoxic-ischemic cerebral damage in fetal rats may trigger the homeostatic control system in the endoplasmic reticulum through the increased expression of GRP78.The apoptotic pathway mediated by caspase-12 in the endoplasmic reticulum may be one of the mechanisms underlying cerebral ischemic injury.

Expression pattern of transforming growth factors observed immunohistochemically in fetal rat / 대한해부학회지

The roles of TGF-beta1, beta2 and beta3 according to gestational ages and histodifferentiation were studied using immunohistochemistry with rat fetuses. 1. From day 14 to 21 of fetal rat, all the TGF-beta1, beta2 and beta3 were expressed in endocardium, myocardium, bronchiole, tunica intima of blood vessles, mucosa and serosa of intestine, striated muscle, hepatic capsule, hepatic hemopoietic cells, meninges, epidermis and dermis. 2. From day 14 to 21 of fetal rat, TGF-beta1 was not expressed in the smooth muscle of blood vessel and intestinal tract, alveolar cell and renal tubular cell. TGF-beta2 was not expressed in the smooth muscle of blood vessel and intestinal tract and alveolar cell. TGF-beta3 was not expressed in osteocyte, alveolar cell, and basement membrane of renal cuboidal epithelial cell. And TGF-betas expressed especially in early or late gestational age and the degree of expression increased or decreased with gestational age. 3. The TGF-beta1, beta2 and beta3 were expressed in tissues originated from 3 germ layers except several tissues, and those originated from mesoderm exhibited strong expression. The TGF-beta1 was expressed more widely than TGF-beta2 and beta3 during gestation. In summary, TGF-beta1, beta2 and beta3 were considered as important control factors of cell to cell interaction in the morphogenesis of tissues during fetal development.

Bone Resorption of Cultured Middle Ear Cholesteatomatous Epithelium-In Vitro Study / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Bone resorption is an important cause of complications of chronic cholesteatomatous otitis media. Authors cultured the calvarial bone of fetal rat and measured the calcium release by keratinocytes and fibroblasts from cholesteatoma and postauricular skin (PAS). MATERIALS AND METHODS: Keratinocytes and subepithelial fibroblasts were cultured from cholesteatoma and PAS. The pregnant rats were sacrificed 24 hours after subcutaneous injection of Ca45 and the fetal calvarial bones were cultured with or without the supernatants of keratinocytes and fibroblast culture. The amount of released radioactive calcium was analyzed using beta-ray counter. RESULTS: The percentage (%) of calcium release was 28.95+/-4.0% in the control and 31.86+/-3.0% in the supernatant of PAS keratinocytes. In the cholesteatomatous keratinocytes, the released calcium was 34.99+/-6.1% and significantly greater than the values of control and PAS. Using the supernatants from the fibroblast culture, cholesteatoma and PAS showed higher calcium release than the control, but there was no significant difference between the two tissues. CONCLUSIONS: Through this study, authors showed the in-vitro bone resorption by cholesteatoma and concluded that cholesteatomatous epithelial cells can be involved in the bone resorption.

In Vitro Effects of Ascorbic Acid on the Repair Process of the Fetal Rat Femur Damaged by Cyclophosphamide / 대한정형외과학회잡지

After bone damage of the fetal rat femurs induced by administrating cyclophosphamide(CP),(1/8 LD50) to the pregnant rat on 13th day of gestation, the effects of serum and ascorbic acid on the repair process of the bone during organ culture were studied, histologically and scanning electron microscopically. CP-damaged fetal femurs harvested at 20 days of gestation were cultured fro 2, 5 and 7 days in the waymouth media(WM) with or without fetal bovine serum(FBS) and ascorbic acid, and were observed with light microscope and JSM-35C scanning electron microscope. The results were as follows:1. CP-damaged bone tissue cultured in WM with 10% FBS showed relatively enhanced activities in the differentiation of chondrocytes and ossificstion as compared to that cultured in WM. 2. CP-damaged bone tissue cultured in WM with 10% FBS and 100µg/ml ascorbic acid, showed increase in the length of the bone marrow cavity, and active formation of new osteoid and collagen bundles. 3. The bone tissues cultured in WM with 10% FBS and 400µg/ml ascorbic acid revealed active deposition of bone matrix, thickening of periosteum and marked elongation of the bone marrow cavity. 4. Bone trabeculae of CP-damaged femurs cultured for 2 days in WM showed poor cell proliferation and insignificant bone matix formation. 5. The number of new cells and the amount of the collagen fibrils increased on the bone trabeculae of the bone cultured in WM with 10% FBS as compared to that cultured in WM and this increase was enhanced as the culture time progressed. 6. A remarkable increase was noted in the number of cells and collagen fibrils in the bone tissues cultured in WM with 10% FBS and ascorbic acid than in those cultured in WM with 10% FBS. 7. The number of the spherules formed by cellular component with collagen fibrils is more numerous than that formed by calcospherites associated with collagen fibrils.

The Effects of Ascorbic Acid on the Growth of the Fetal Rat Bone in Ca++ / 대한정형외과학회잡지

In order to study the effect of ascorbic acid on the growth of the fetal rat long bones in calcium free culture medium, fetal femurs from rat fetus on 19th day of gestation were cultured for 1, 3, 5, 7 and 9 days in medium described below. Culture media used were MEM, Ca++

Effect of 3,3',5-trliodothyronine on the expression of Go?in primary cultured neurons of rats / 中华内分泌代谢杂志

Objective To investigate the effect of triiodothyronine on the expression of alpha subunit of C protein Co (Coa) gene in cultured rat cerebral cortex neurons. Methods Primary cultured neurons of rat were prepared from the cerebral cortex at embryonic day 19. The neurons were cultured in DMEM supplemented with 15% fetal calf serum. After 7 days, the neurons were cultured in different media; DMEM supplemented with 15% fetal calf serum (group A), DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group B), 0.5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group C) , 5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group D) and 50nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group E). The neurons were cultured for another 7 days and total RNA was extracted. Goa mRNA leves were measured by competitive RT-PCR. Results As compared with group A, Coa mRNA leves in group B, D and E were low (P 0.05). Conclusion T, has dual effects on the expression of Coa gene in cultured rat cerebral cortex neurons. T3 down-regulated Coa mRNA in cultured neuron. However, in the absence of T3, Goa mRNA was also very low. Thyroid hormone may exert their action on brain development by regulating the the expression of Goa gene.

Adrenal allografts using fetal rats as donors / 第三军医大学学报

Objective To study the reconstruction and secretory function of the grafts after adrenal transplantation using fetal rats as donors.Methods Thirty adult SD rats randomly received sham operation(group A),excision of bilateral adrenals(groups B,C,D and E),and additionally adrenal transplantation in groups C,D and E using fetal rats aged 16,18 and 20 days respectively as donors.Results The weight of group A got down to the lowest at the 3rd week,and restored to the preoperation level at the 4th week after sham operation.The weight of group B gradually rose up within 4 weeks.The weight of groups C and D rose up in the first 2 weeks,then down.The weight of group E rose slowly within the first 3 weeks,then down.The serum level of corticosterone decreased sharply in group B after operation,while that of groups C,D and E went down obviously in the first 2 weeks,then up,but that in group C was significantly lower than that in groups D and E.Microscopically,we found the capsule of allografts thicken and underneath many proliferative cells and well-constructed glomerular and fascicular zone 4 weeks after operation.Conclusion After transplantation,the fetal adrenal could survive and reconstruct well,and its secretory function gets better gradually.The graft from the elder donor is better than that from the younger in reconstruction and secretory function,but shows no obvious difference in immunological rejection.

STUDY ON THE EXPERIMENTAL CONDITIONS OF BMG-INDUCED CHONDROCYTE FORMATION IN VITRO / 西安交通大学学报(医学版)

Thigh skeletal muscle of 19d～21d(near-term) fetal rat cultured together with bone matrix gelatin (BMG) in vitro. The medium was DMEM containing 3700 mg/L NaCHO_3. After 1O d in culture, there were BMG-induced chondrocytes in 7 out of 8 explants. After 15、20 and 25 d in culture. BMG-induced chondrocytes were observed in all of the 30 expiants(10 explants in each point of time-period). The result shows that the experimental conditions and culture medium used in this paper were suitable for the system of BMG-induced choudrocyte formation in vitro.

EFFECT OF VELVET ANTLER POLYPEPTIDE(VAP) ON DIFFERENTIATION OF RAT BRAIN-DERIVED STEM CELLS IN VITRO / 解剖学报

Objective To investigate the probabilities of brain-derived stem cells from fetal rats differentiating into neurons and astrocytes by velvet antler polypeptide(VAP) in vitro. Methods Neural stem cells from E12-14d rats were cultured for 7 days until neural stem cells (NSCs) aggregations were formed into neurospheres. The neurospheres were cultured at different concentrations of VAP, and immunocytochemistry was used to detect the differentiation of neural stem cells. Results The differentiated cells in 50?g/L VAP group are more than that in control group; the number of NSE positive cells in 50?g/L,100?g/L and 200?g/L groups is more than that in control group.Conclusion Neural stem cells can be successfully induced into neurons by VAP in vitro, which could provide a basis for regeneration of nerve system.;