Assessment of Awareness and Understanding of Hemolytic Disease of the Fetus and Newborn in the Beninese Population

Background: Hemolytic Disease of the Fetus and Newborn (HDFN) arises from blood group incompatibility, especially the RhD antigen. In Benin, systematic ABO RhD blood grouping is poorly understood by many midwives and nurses. Nearly one in ten women risk having children with HDFN. This study aimed to determine the level of knowledge of the Beninese population on HDFN. Methods: Data were collected from June 2023 to March 2024. Participants completed a Kobotoolbox questionnaire on WhatsApp, with in-person assistance for illiterate participants. The study involved 521 participants from across Benin. Data were analyzed using SigmaPlot version 14.0. Results: Among the 521 participants, 298 were women (57.20%) aged 18 to 77 years. The majority (40.69%) were aged 26 - 35. Over a third (35.51%) did not know their RhD blood group. Most (59.12%) were unaware of the risks for RhD discordant couples. Among those with a partner, 25.16% were in at-risk couples for HDFN, and over half (59.12%) were unaware of this risk. There was no significant association between being in a high-risk union and knowledge of the risk or education level. Conclusion: Only 40.88% of the Beninese population are aware of HDFN, indicating a low level of knowledge.

Nerve protective effect of Baicalin on newborn HIBD rats

Objetive:To investigate the nerve protective effect and mechanism of baicalin on newborn rats with hypoxic ischemic brain damage(HIBD).Methods:A total of 64 SD newborn rats were randomlu divided into control group.model group.nerve growth factor group and baicalin group.with 16 in each group.Left carotid artery ligation method was adopted to establish the HIBD model except fou in control group,which was treatde with intraperitoneal injection of salin e10mL/kg for 3 d.After oxygen recovery on hypoxia ischemia rats.intraperitoneal injectionof salin 10mL/kg was adopted in model group for 3 d.Intraperitoneal injection of nerve growth factor injection50μg/kg per day was adopted in nerve growth factor group for 3 d:intraperitoneal injection of radix scutellariae 16mg/kg per day was adopted in baicalin group for 3 d after modeeling.Four rats of each group were sacrificed at Day 1,2,3,7 for microscopic observation of pathological morphological changes in brain tissus aften HE staining,S-P immunohistochemical method was used for observation of Fas and FasL expression in brain cells.Results:Neat structure of cells was observed in control group;edema cells in disordered arrangement was observed in model group,with some cells necrosis and cavity change;tissue injury in nerve growth factor group and baicalin group was significantly lighter than that in model group;Fas and FasL expression in model group,nerve growth factor group and baicalin group were significantiy higher than that in control group at different time points(P<0.05):Fas and FasL expression in nerve growth factor group and baicalin group were significantly lower than that in model group at different time points(P<0.05):There was no statistical diggerence of Fas,FasL expression at each time point between nerve growth factor group and baicalin group(P>0.05).Conclusions:Baicalin can reduce expression of Fas and FasL in HIBD rats,inhibit apoptosis of nerve cells,thus achieve the protective effect on HIBD rat nerves.

Culture of Motor Neurons from Newborn Rat Spinal Cord

A protocol for the isolation, purification and culture of motor neurons from newborn rat spinal cord was described and the effect of glial cell line-derived neurotrophic factor （GDNF） on the growth of neurite of motor neurons was investigated in vitro. Spinal motor neurons （SMNs） were dissociated from ventral spinal cord of postnatal day 1 rats. The culture system for SMNs was established by density gradient centrifugation, differential adhesion, and use of serum-free defined media and addition of exogenous GDNF. After 72-h culture, the cells displayed the characteristic morphology of motor neurons, exhibited extensive neuritic processes and were positive for choline acetyl- transferase （CHAT） expression. The neurite length of SMNs in GDNF groups was significantly longer than that in control group （P〈0.05）. This protocol can be adapted for various postnatal motor neurons studies.

Environmental enrichment promotes neural remodeling in newborn rats with hypoxic-ischemic brain damage

We evaluated the effect of hypoxic-ischemic brain damage and treatment with early environmental enrichment intervention on development of newborn rats, as evaluated by light and electron microscopy and morphometry. Early intervention with environmental enrichment intelligence training attenuated brain edema and neuronal injury, promoted neuronal repair, and increased neuronal plasticity in the frontal lobe cortex of the newborn rats with hypoxic-ischemic brain damage.

Interventional effect of laser acupoint radiation on the expression of Nissl body and brain-derived neurotrophic factor in newborn rat models with ischemic/hypoxic cerebral injury

BACKGROUND： Some researches report that He-Ne laser can activate function of erythrocytes and increase content of blood and oxygen via bio-stimulating effect; therefore, it suspects that laser radiation at Baihui and Dazhui can partially increase blood circulation for oxygen-supplying content of brain and improve functional status of neurons. OBJECTIVE： To verify the effects of laser radiation at Baihui and Dazhui on the expression of Nissl body of brain tissue neurons and brain-derived neurotrophic factor （BDNF） in newborn rats with ischemic/hypoxic cerebral injury. DESIGN： Randomized controlled animal study. SETTING： Department of Neurological Histochemistry, Xianning University. MATERIALS： Forty Wistar rats of 7 - 8 days old, weighing 15 - 20 g and of both genders, were selected from Wuhan Experimental Animal Center. All the rats were randomly divided into sham operation group （n =8）, model group （n =16） and radiation group （n = 16）. The experimental animals were disposed according to ethical criteria. BDNF kit was provided by Wuhan Boster Bioengineering Co., Ltd. METHODS： The experiment was carried out in the Department of Neurological Histochemistry, Xianning University from April 2005 to October 2006. Rats in the radiation group and model group were performed with ligation of left common carotid artery, recovered at room temperature for 1 - 6 days, maintained in self-made hypoxic cabin under normal pressure and injected mixture gas （0.05 volume fraction of 02 and 0.92 volume fraction of N2） for 2 hours. In addition, rats in the sham operation group were treated with separation of left common carotid artery but not ligation and hypoxia. Rats in the model group were not given any treatment; while, rats in the radiation group were exposed with He-Ne laser of 63.28 nm in the wave length at Baihui and Dazhui acupoints on the second day after ischemia-hypoxia. The radiation was given for 10 minutes per day and once a day. Ten days were regarded as a course and the rats were exposed for 2 courses in total. At 20 days after routine breeding, left hemisphere tissues of rats in the three groups were collected for staining of Nissl body and immunohistochemistry of BDNF. MAIN OUTCOME MEASURES： Nissl body staining in left hemisphere tissue and expression of immune positive cells of BDNF. RESULTS： All 40 rats were involved in the final analysis. （1） Nissl body staining： Neuronal cytoplasm of brain tissue was full of blue granule Nissl bodies in the sham operation group; while, Nissl body in neuronal cytoplasm in the model group was stained slightly and had a certain degree of degeneration; meanwhile, there were a lot of blank area in ischemic region. Nissl body in neuron cytoplasm was gradually recovered in the radiation group and relieved as compared with that in the model group. （2） Positive cells of BDNF： Number of immune positive cells of BDNF which were ligated in lateral cerebral hemisphere of rats in the model group was higher than that in the sham operation group （P 〈 0.05）; while, BDNF expression in the radiation group was increased as compared with that in the model group （P 〈 0.05）. CONCLUSION： After laser acupoint radiation, Nissl body is increased and BDNF expression is also increased. This suggests that laser acupoint radiation has neuroprotective effect on brain tissue after ischemia-hypoxia injury.

Effect of Retinoic acid on Platelet-derived Growth Factorand Lung Development in Newborn Rats

Summary: The influence of platelet-derived growth factor (PDGF) on lung development in newborn rats and the effect of retinoic acid (RA) on PDGF in lung development were investigated. Newborn Sprague-Dawley (SD) rats were randomly assigned to two groups: control group and RA group. The rats in RA group was intraperitoneally injected with all trans-retinoic acid (500 μg/kg every day) for consecutive 3 days after birth, while those in the control group were not subjected to intervention. Immunohistochemical assay was performed to locate the expression of PDGF. mRNA levels of PDGF were measured by reverse transcription polymerase chain reaction (RT-PCR) at age of 1, 3, 5, 7, 10, 14, 21 days. The method of radial alveolar counts (RAC) was used to measure the amount of the alveoli of the lungs. It was found that with increasing days, levels of PDGF-A and PDGF-B changed to verying degrees. RA could elevate significantly the expression levels of PDGF-A mRNA and protein (P<0.01), but not affect the expression levels of PDGF-B mRNA and protein markedly (P>0.05). It is suggested that PDGF might play an important role in lung development. RA can stimulate lung development through increasing the expression levels of PDGF-A mRNA and protein.

THE DISTRIBUTION OF ATRIONATRIURETIC POLYPEPTIDE (ANP)-LIKE SUBSTANCE IN THE HEART OF THE HUMAN FETUS AND RAT-AN IMMUNOHISTOCHEMICAL STUDY

human fetuses and 15 SD rats were used in this study. The distribution of theANPlike substance in the hearts of human fetuses and rats was studied. The results were as follows:①In human fetus ANP-like substance was found throughout in the atrial wall, but more intensely in the inner layer. It lacked in the ventricular wall. ②In young rat atrum, ANP-like substance wasdistributed over the whole wall, but richer in the cells beneath the endocardium and epicardium. In adult rat atrium, distribution of ANP-like substance was more intensely beneath the epicardium than that near the lumen. In the ventricle, ANP-like substance was found only in the young rat, especially on the left side of interventricular septum (IVS). So the distribution of ANP in rat hearts changes with age. ③In the sinus node (SAN) and atrioventricular node (AVN) of both human fetuses and rats showed a small amount of ANP-like substance, and more in peripheral part of the nodes, but thereactive intensity was weaker than that of the atrial wall. The presence of ANP in conduction system may have some functional significance to influence the pulse conduction.

Changes in hippocampal neurons and memory function during the developmental stage of newborn rats with hypoxic-ischemic encephalopathy

BACKGROUND: Under the normal circumstance, there exist some synapses with inactive functions in central nervous system (CNS), but these functions are activated following nerve injury. At the early stage of brain injury, the abnormal functions of brain are varied, and they have very strong plasticity and are corrected easily. OBJECTIVE: To observe the changes of neuronal morphology in hippocampal CA1 region and memory function in newborn rats with hypoxic-ischemic encephalopathy(HIE) from ischemia 6 hours to adult. DESIGN: Completely randomized grouping, controlled experiment. SETTING: Taian Health Center for Women and Children; Taishan Medical College. MATERIALS: Altogether 120 seven-day-old Wistar rats, of clean grade, were provided by the Experimental Animal Center, Shandong University of Traditional Chinese Medicine. Synaptophysin (SYN) polyclonal antibody was provided by Maixin Biological Company, Fuzhou. METHODS: This experiment was carried out in the Laboratory of Morphology, Taishan Medical College between October 2000 and December 2003. ① The newborn rats were randomly divided into 2 groups: model group and control group, 60 rats in each group. Five rats were chosen from each group at postoperative 6 hours, 24 hours, 72 hours, 7 days, 2 weeks and 3 weeks separately for immunohistochemical staining. Fifteen newborn rats were chosen from each group at postoperative 4 weeks and 2 months separately for testing memory ability (After test, 5 rats from each group were sacrificed and used for immunohistochemical staining)② The right common carotid artery of newborn rats of model group was ligated under the anesthetized status. After two hours of incubation, the rats were placed for 2 hours in a container filled with nitrogen oxygen atmosphere containing 0.08 volume fraction of oxygen, thus, HIE models were created; As for the newborn rats in the control group, only blood vessels were isolated, and they were not ligated and hypoxia-treated. ③ Thalamencephal tissue sections of newborn rats of two groups were performed DAB developing and haematoxylin slight staining. Cells with normal nucleous in 250 μm-long granular layer which started from hippocampal CA1 region were counted with image analysis system under high-fold optical microscope (×600), and the thickness of granular layer was measured. The absorbance (A) of positive reactant of SYN in immunohistochemically-stained CA1 region was measured. Learning and memory ability were measured with step through test 3 times successively. ④ t test and paired t test were used for comparing intergroup and intragroup difference of measurement data respectively, and Chi-square for comparing the difference of enumeration data. MAIN OUTCOME MEASURES: Comparison of cytological changes in hippocampal CA1 region and memory ability at different postoperative time points between two groups. RESULTS: Totally 120 newborn rats were involved in the result analysis. ① Cell morphological changes in hippocampal CA1 region: In the control group, with aging, perikaryon, nucleus and nucleolus in cortex of parietal lobe were significantly increased, Nissl body was compacted, the amount of neurons was declined, but the A of SYN positive reactant was relatively increased. In the model group, at postoperative each time point, neurons were seriously shrunk and dark-stained, nucleus was contracted, chromatin was condensed, nucleolus was unclear, even cells disappeared, especially the cells in 6 hours and 24 hours groups. The amount of neurons with normal morphology in hippocampal CA1 region and granular layer thickness in the model group at postoperative each time point were significantly less or smaller than those in the control group at postoperative 6 hours respectively (t =3.002-1.254, P < 0.01). The A value of SYN positive reactant at postoperative 2, 3 and 4 weeks was significantly higher than that at previous time point (t =2.011-2.716,P < 0.05-0.01). ② Test results of learning and memory ability: In the first test, there was no significant difference in the ratio of rats which kept memory ability between two groups (P > 0.05); In the third test, the ratio of rats which kept memory ability in the model group was significantly lower than that in the control group at postoperative 4 weeks and 2 months[53%(8/15),100%(15/15);60%(9/15),93%(14/15),χ 2=2.863,2.901,P < 0.01]. CONCLUSION: The destroyed hippocampal structure induces the decrease of learning and memory ability of developmental rats. Early interference can increase the quality of neurons and also promote functional development of the nervous system.

抗-M致胎儿新生儿溶血病实验室检测及预防策略分析

目的分析血清学检测结果在抗-M致胎儿新生儿溶血病(HDFN)诊断和治疗中的应用,探讨HDFN预防策略。方法对2017年1月—2023年12月本实验室确诊的12例抗-M引起的HDFN血清学检测结果进行回顾性分析,包括母亲和患儿的血型鉴定,血清总胆红素、血红蛋白、抗体效价检测,新生儿溶血3项实验。收集患儿和母亲的临床资料,包括妊娠史、输血史、产前抗体检测、宫内输血史、分娩孕周,并随访患儿预后。结果12例抗-M引起的HDFN中,患儿母亲均为RhD+NN表型,患儿均为RhD+MN表型,母子MN血型均不相容。在ABO血型系统中,母子ABO血型不相容占41.7%(5/12)。患儿母亲皆无输血史,在4℃检测效价中位数32,37℃效价中位数4。3例患儿母亲有多次宫内输血史,发生率为25%(3/12)。1例为首胎妊娠异常,占比8.3%(1/12),7例为异常妊娠流产,不良孕产史发生率58.3%(7/12)。6例患儿母亲早产,占比50%(6/12)。3例患儿母亲定期产检,并鉴定抗体特异性,占比25%(3/12)。12例患儿游离抗体在4℃检测效价中位数6,37℃效价中位数2。2例患儿抗-M在37℃检测无反应,阴性率16.7%(2/12)。患儿直抗阳性率8.3%(1/12),放散阳性率16.7%(2/12)。血红蛋白最低值中位数75 g/L,12例患儿均接受了输血治疗。总胆红素峰值中位数157.5μmol/L,均未达到换血阈值。患儿迟发性贫血率16.7%(2/12),患儿出生后死亡率8.3%(1/12),11例患儿预后无生长和神经发育迟缓情况。结论抗-M能引起严重的HDFN,也可发生于第1胎,抗体效价强度与疾病的严重程度不相关,容易引起迟发性贫血,应根据抗-M血清学特征和临床症状定期监测,及时干预治疗。

单核细胞单层试验在预测IgG抗-M相关胎儿新生儿溶血病中的应用

目的探讨单核细胞单层实验(monocyte monolayer assay,MMA)是否能够用于IgG抗-M相关胎儿新生儿溶血病(hemolytic disease of fetus and newborn,HDFN)的预测。方法选取8例含有IgG抗-M的孕妇并采集血浆标本,其中有胎儿水肿等严重临床症状及无明显临床症状的各4例;8份血浆用二硫苏糖醇(dithiothreitol,DTT)灭活,与M抗原阳性红细胞孵育致敏后,将致敏红细胞与单核细胞混合进行吞噬试验,同时设立阳性及阴性对照,并计算吞噬率;采用t检验对2组吞噬率进行比较。结果4例发生严重抗-M相关HDFN的孕妇的MMA吞噬率分别为15.37%、13.05%、9.17%和24.50%,均值为15.52%;检出IgG抗-M但未发生HDFN的孕妇,其MMA吞噬率分别为8.74%、11.07%、5.12%和6.23%,均值为7.79%,2组吞噬率无差异(P>0.05)。2组吞噬率分别与阴性对照比较均无差异(P>0.05),但均明显低于阳性对照(P<0.05)。结论IgG抗-M介导单核细胞吞噬的能力较低,提示抗-M导致胎儿水肿的机制可能并非红细胞被吞噬破坏而发生的溶血,因此体外单核细胞单层实验可能不适用于IgG抗-M相关HDFN的预测。对于检出IgG抗-M的孕妇,现仍需通过定期监测胎儿大脑中动脉血流,来判断胎儿宫内贫血情况。

罕见抗-Di^(b)致严重胎儿新生儿溶血病的实验室检测与相关研究

目的对1例高频抗体导致的胎儿新生儿溶血病(hemolytic disease of the fetus and newborn,HDFN)进行检测、鉴定及配血。方法对患儿进行新生儿溶血试验,对母亲进行血清学意外抗体鉴定,并对母亲红细胞进行常见高频抗原鉴定;对检出抗体进行IgG分型检测,并用流式细胞术进行单核细胞体外吞噬致敏红细胞试验,以检测抗体相关的吞噬率;对患儿母亲、父亲及舅舅进行相关红细胞血型基因测序;利用稀释的母亲血浆和抗人球卡法,在献血者中进行大规模相合血液的筛选。结果产妇鉴定为Di(b-)稀有血型,产生了抗-Di b(效价512)并导致了严重的HDFN;抗-Di b亚型分型为IgG1和IgG2型,单核细胞体外吞噬效率为88.83%(74.7/84.09);产妇亲属中没有相合献血者,后续从5520名献血者中筛选到2例Di(b-)相合血液,患儿接收输血治疗后康复出院。后续在51334名献血者中筛查到17名Di(b-)献血者,该数据表明Di(b-)在广州地区献血者中的分布频率约为三千分之一(0.033%,17/51334)。结论综合利用血型血清学及分子生物学方法诊断了抗-Di b所致的严重HDFN,建立了1种有效大规模筛查Di(b-)稀有血型的方法并找到相合血液,为建立Di(b-)稀有血型库奠定了基础。

RhD阴性孕产妇与HDFN发生的相关性及影响因素分析

目的 通过对RhD阴性孕产妇腹中胎儿和分娩的新生儿发生胎儿新生儿溶血病(hemolytic disease of the fetus and newborn,HDFN)的相关指标对比分析,为预防和治疗HDFN提供参考和指导。方法 收集我院2018年1月—2022年12月分娩的RhD阴性孕产妇737名,比较新生儿是否发生RhD血型不合、ABO血型不合导致的HDFN及其影响因素,发生RhD-HDFN和发生ABO-HDFN的相关影响因素。分析发生RhD-HDFN和发生ABO-HDFN患儿的实验室指标差异;分析IgG抗-D效价≤16和≥32的孕产妇分娩的新生儿发生RhD-HDFN的实验室指标差异。结果 737名RhD阴性孕产妇中,发生RhD-HDFN的母婴ABO血型相同或相容者比率88.89%(40/45)显著高于母婴ABO血型不相容者11.11%(5/45)。母体二次妊娠及以上发生RhD-HDFN比率93.33%(42/45)显著高于ABO-HDFN 60.66%(37/61)者。母体IgG抗-D效价≥32者分娩的新生儿血红蛋白(hemoglobin,Hb)最低值低于母体IgG抗-D效价≤16者(χ^(2)=5.61,P<0.05),母体IgG抗-D效价≥32者分娩的新生儿血清总胆红素(total bilirubin,TBil)峰值高于IgG抗-D效价≤16者(χ^(2)=4.471,P<0.05)。结论 RhD阴性孕产妇中,母婴ABO血型相同或相容及孕产次≥2者,相应新生儿更易发生RhD-HDFN,母体IgG抗-D效价≥32者发生新生儿溶血的严重程度显著高于抗-D效价≤16者。

石菖蒲挥发油对宫内感染所致新生鼠肺损伤的作用

目的探讨石菖蒲挥发油(volatile oil of Acorus tatarinowii,VOAT)对宫内感染所致新生鼠肺损伤的作用及可能的机制。方法随机选取6只妊娠15 d的SD大鼠作为对照组,子宫颈内注射大肠埃希菌构建宫内感染大鼠模型,并随机分为模型组及VOAT低、中、高剂量组,每组6只,次日给药,其中VOAT低、中、高剂量组大鼠分别灌胃VOAT 5、10、20 g·kg^(−1)·d^(−1),模型组和对照组大鼠灌胃等量生理盐水,连续给药直至分娩。记录各组新生鼠胎龄;测定新生鼠肺指数;用酶联免疫吸附试验(enzymelinked immunosorbent assay,ELISA)测定新生鼠血清白细胞介素(interleukin,IL)-21和IL-6、IL-1β水平;苏木素伊红(hematoxylin-eosin,HE)染色检测新生鼠肺组织病理学变化;实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,RT-qPCR)检测肺组织中IL-21、IL-21R mRNA的相对表达水平;Western blotting检测肺组织中IL-21、IL-21R、STAT3、p-STAT3蛋白的相对表达量。结果与对照组比较,模型组及VOAT低、中、高剂量组新生鼠胎龄缩短,肺指数降低,血清IL-21、IL-6及IL-1β水平,肺组织中IL-21、IL-21R mRNA的相对表达量及IL-21、IL-21R和p-STAT3蛋白的相对表达量均升高(P<0.05),并出现不同程度的炎性细胞浸润、肺泡壁增厚、肺泡腔变大、结构紊乱等病理学变化;与模型组比较,VOAT低、中、高剂量组新生鼠肺指数升高,血清IL-21、IL-6及IL-1β水平,肺组织中IL-21、IL-21R mRNA的相对表达量及IL-21、IL-21R和p-STAT3蛋白的相对表达量均降低(P<0.05),肺组织病理损伤有不同程度减轻;VOAT对宫内感染所致新生鼠肺损伤的作用效果呈剂量依赖性(P<0.05)。结论VOAT可减轻宫内感染所致新生鼠肺损伤,其作用机制可能与抑制IL-21/STAT3通路激活、降低机体炎症水平有关。

改良56℃热放散法与冷冻复融放散法对ABO型新生儿溶血病的临床应用价值比较

目的比较改良56℃热放散法和冷冻复融放散法对检测ABO型新生儿溶血病(ABO-HDFN)的阳性检出率、试验成本等方面进行比较,探讨两种方法的临床应用价值。方法选择2023年3月本院接收的所有进行新生儿/脐带血标本进行ABO溶血病检测(直接抗人球蛋白试验、游离抗体试验、放散试验),其放散试验时同时进行改良56℃热放散和冷冻复融放散,用统计学方法对两种放散方法结果进行比较分析。结果共纳入非O型的新生儿/脐带血标本256例进行ABO-HDFN检测,其中A型145例,B型109例,AB型2例。改良56℃热放散法的阳性检测率69.92%,冷冻复融放散法的阳性检出率为64.45%,二者无统计学差异。比较两种方法在不同血型、不同凝集强度、不同直接抗人球蛋白试验结果(阴性/阳性)情况下的放散结果,二者均无统计学差异;仅游离抗体结果为阳性时,两种方法具有统计学差异。结论在大多数情况下两种放散方法对于ABO-HDFN的诊断价值无统计学差异。各实验室可以结合临床患者的紧急程度以及本实验室的仪器设备、耗材配置、工作人员经验和时间成本等综合考虑,选择适合本实验室的方法。

Role of neuronal nitric oxide synthase and inducible nitric oxide synthase in intestinal injury in neonatal rats

AIM： TO investigate the dynamic change and role of neuronal nitric oxide synthase （nNOS） and inducible nitric oxide synthase （iNOS） in neonatal rat with intestinal injury and to define whether necrotizing enterocolitis （NEC） is associated with the levels of nitric oxide synthase （NOS） in the mucosa of the affected intestine tissue. METHODS： Wistar rats less than 24 h in age received an intraperitoneal injection with 5 mg/kg lipopolysaccharide （LPS）. Ileum tissues were collected at 1, 3, 6, 12 and 24 h following LPS challenge for histological evaluation of NEC and for measurements of nNOS and iNOS. The correlation between the degree of intestinal injury and levels of NOS was determined. RESULTS： The LPS-injected increase in injury scores pups showed a significant versus the control. The expression of nNOS protein and mRNA was diminished after LPS injection. There was a negative significant correlation between the nNOS protein and the grade of median intestinal injury within 24 h. The expression of iNOS protein and mRNA was significantly increased in the peak of intestinal injury. CONCLUSION： nNOS and iNOS play different roles in LPS-induced intestinal injury. Caution should be exerted concerning potential therapeutic uses of NOS inhibitors in NEC.

Dynamic change of epidermal growth factor in neonatal rat with intestine injury

AIM: To determine whether diminished levels of epidermal growth factor (EGF) were present in neo-natal rats with intestinal injury and related with the degree of intestinal injury, so we modeled a model in neo-natal rats of intestinal injury and to examine the dynamic levels of EGF on injury of intestine.METHODS: One-day-old Wistar rat pups received an intraperitoneally injection with 4 mg/kg lipopolysaccharide (LPS), followed by collection of ileum tissue at 1, 3, 6, 12,and 24 h following LPS administration. The ileum was for histological evaluation of NEC and for measurements of EGF using ABC-ELISA. The correlation between the degree of intestinal injury and levels of EGF was determined. RESULTS: The LPS-injected pups also showed a significant increase in injury scores at 1, 3, 6, 12, and 24 h [respectively, (1.08±0.61), (1.63±0.84), (1.95±0.72), (2.42±0.43)and (2.21±0.53)] vsthe control (0.12±0.17) (P＜0.01).EGF levels at 1, 3, 6, 12 h [respectively, (245.6±49.0), (221.4±39.0), (223.4±48.1), (246.0±46.6)] pg/mg were significantly loss than the control (275.6±50.4) pg/mg (P＜0.05). There was a significant negative correlation between the EGF levels and the grade of intestinal injury within 24 h (P＜0.05).CONCLUSION: Neo-natal rats with intestinal injury have significantly lower levels of ileum EGF. Reduced levels of this growth factor might be related to the pathogenesis of NEC.

Expression of estrogen receptor (ER) -α and -βtranscripts in the neonatal and adult rat cerebral cortex, cerebellum, and olfactory bulb

In the present study expression of estrogen receptor subtype -alpha (ERalpha) and -beta (ERbeta) in the cerebral cortex, cerebellum, and olfactory bulb was investigated and compared between neonatal (1 to approximately 3-days-old) and adult (250 to approximately 350 g) rats, using reverse transcription-polymerase chain reaction (RT-PCR). No ERalpha transcripts were detectable in the adult cerebellum and olfactory bulb, whereas very weak expression of ERalpha was present in the adult cerebral cortex. No significant difference in ERbeta transcripts was detectable between the neonatal and adult rats. While transcripts for both ER subtypes were co-expressed in these brain areas of neonatal rats, although ERalpha expression was significantly weaker than ERbeta. Even in the cerebral cortex known to contain both ER subtypes in adult rats, ERalpha transcripts in neonatal rats were much higher than in adult. These observations provide evidence for the existence of different expression patterns of ERalpha/ERbeta transcripts in these three brain areas between the neonatal and adult rats, suggesting that each ER subtype may play a distinct role in the regulation of differentiation, development, and functions of the brain by estrogen.

Protective effect of insulin and glucose at different concentrations on penicillin-induced astrocyte death on the primer astroglial cell line

Astrocytes perform many functions in the brain and spinal cord.Glucose metabolism is important for astroglial cells and astrocytes are the only cells with insulin receptors in the brain.The common antibiotic penicillin is also a chemical agent that causes degenerative effect on neuronal cell.The aim of this study is to show the effect of insulin and glucose at different concentrations on the astrocyte death induced by penicillin on primer astroglial cell line.It is well known that intracranial penicillin treatment causes neuronal cell death and it is used for experimental epilepsy model commonly.Previous studies showed that insulin and glucose might protect neuronal cell in case of proper concentrations.But,the present study is about the effect of insulin and glucose against astrocyte death induced by penicillin.For this purpose,newborn rat brain was extracted and then mechanically dissociated to astroglial cell suspension and finally grown in culture medium.Clutters were maintained for 2 weeks prior to being used in these experiments.Different concentrations of insulin（0,1,3 nM）and glucose（0,3,30 mM）were used in media without penicillin and with 2 500μM penicillin.Penicillin decreased the viability of astroglial cell seriously.The highest cell viability appeared in medium with 3 nM insulin and 3 mM glucose but without penicillin.However,in medium with penicillin,the best cell survival was in medium with 1 nM insulin but without glucose. We concluded that insulin and glucose show protective effects on the damage induced by penicillin to primer astroglial cell line.Interestingly,cell survival depends on concentrations of insulin and glucose strongly.The results of this study will help to explain cerebrovascular pathologies parallel to insulin and glucose conditions of patient after intracranial injuries.

Development of the Gusto Facial Reflex in Perinatally Undernourished Rats

In the rat, the taste system plays a critical role in motivating the animal to consume nutrients and avoid toxic substances. In neonatal rat the orofacial movements can be modified by the application of sucrose and quinine in the mouth, producing ingestion or rejection responses, respectively, but there is no information available on the gusto facial reflexes (GFR) under perinatal fasting. The aim of the present study was to determine how undernutrition can affect the development of orofacial responses to sucrose, quinine, and NaCl during early development. Pregnant dams were undernourished by being given 50% of a balanced diet from G6 to G12, 60% from G13-G18, and 100% from G19-G21. On postpartum days 0 - 9, prenatally underfed (UG) pups continued the undernourishment by remaining for 12 h with a foster dam and 12 h with a nipple-ligated mother. Stimuli were presented as a single droplet of sucrose (S), sodium chloride (NaCl), quinine hydrochloride (Q), and water (W) onto the lips at 1, 3, 5, 7, and 9 days of age, and mouth-opening frequency (MO) and lip-licking frequency (LL) were analyzed. The results show consistent effects associated with age, but not with diet, except in the cases of 0.01 M Q, which provoked in the UG subjects increased MO and the reduction of LL, and 0.1 M Q, which consistently reduced both responses. Moreover, S provoked increases in both MO and LL at 0.1 M while NaCl and water elicited minimal effects on GFR. These data suggest that perinatal undernutrition affects the basic components of the gustatory system necessary to produce GFR in the first postnatal days by possibly interfering with the integration of taste input for food learning and the hedonic aspects of gustatory cues.

Batroxobin Against Anoxic Damage of Rat Hippocampal Neurons in Culture:Morphological Changes and Hsp70 Expression

Batroxobin,the thrombin-like enzyme,is used for therapeutic defibrination. We have found that batroxobin has good therapeutic effect in ischemic reperfusion rats and clinical practices in vivo. But we have not studied the neuroprotective effect of batroxobin on anoxic hippocampal neurons in vitro. The purpose of this study was to obtain further information on the mechanism of the batroxobin-induced neuroprotection and examine the neuroprotective effect on neurons exposed to anoxia. The effect of batroxobin on anoxic damages in cultured hippocampal neurons of neonatal rats was investigated by using morphological changes and heat shock protein 70Kd (Hsp70) immunoreactive expression as indicators. The results indicate that batroxobin, besides its defibrination, may have a direct neuroprotective effect on anoxic damage of hippocampal neurons.