Transformation of hepatitis B serologic markers in babies born to hepatitis B surface antigen positive mothers

AIM:To better understand the clinical significance of hepatitis B seroiogic markers in babies born to hepatitis B surface antigen (HBsAg) positive mothers, the incidence of maternal seroiogic markers of hepatitis B via placenta and its transformation in these babies were investigated. METHODS: Mothers with positive HBsAg were selected in the third trimester of pregnancy. Their babies received immunoprophylaxis with hepatitis B immunoglobulin and hepatitis B vaccine after birth, and were consecutively followed up for hepatitis B seroiogic markers and HBV DNA at birth, mo 1, 4, 7, 12, and 24. RESULTS: Forty-two babies entered the study, including 16 born to hepatitis B e antigen (HBeAg)-positive HBsAg carrier mothers and 26 to HBeAg-negative HBsAg carrier mothers. Apart from four babies born to HBeAg-positive carrier mothers and demonstrated persistent positive HBeAg eventually became HBV carriers, all other babies developed anti-HBs before 12 mo of age. Among the other 12 babies born to HBeAg-positive carrier mothers, HBeAg was detected in 7 at birth, in 4 at mo 1, and in none of them thereafter. No antibody response to the transplacental HBeAg was detected. Among the babies born to HBeAg-negative carrier mothers, anti-HBe was detected 100% at birth and mo 1, in 88.5% at mo 4, in 46.2% at mo 7, in 4.2% at mo 12 and none in mo 24. Among all the immunoprophylaxis-protected babies born to either HBeAg-positive or HBeAg-negative carrier mothers, anti-HBc was detected in 100% at birth, mo 1 and mo 4, in 78.9% at mo 7, in 36.1% at mo 12 and in none at mo 24. CONCLUSION: HBeAg can pass through human placenta from mother to fetus and become undetectable before 4 mo of age, but no antibodies response to the transplacental HBeAg can be detected till mo 24 in the immunoprophylaxis-protected babies. The sole existence of anti-HBe before 1 year of age or anti-HBc before 2 years of age in babies born to HBsAg carrier mothers may simply represent the transplacental maternal antibodies, instead of indicators of HBV infection status.

Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721

AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells. METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SW), were constructed by ligating into the vector, pSupperssorNeo (pSuNeo) digested with restriction enzymes Xba I and Sail and the designed double-chain RNAi primers. A cell model of SMMC-7721 after treatment with RNAi was prepared by transfecting SMMC-7721 cells with the lipofectin transfection method. Strept-avidin-biotin-complex (SABC) immunohistochemical staining and RT-PCR were used to detect survivin gene expressions in SMMC-7721 cells. Flow cytometry was used for the cell cycle analysis. Transmission electron microscopy was performed to determine whether RNAi induced cell apoptosis, and the method of measuring the cell growth curve was utilized to study the growth of SMMC-7721 cells before and after treatment with RNAi. RESULTS: The eukaryotic expression vector of survivin gene RNAi and pSuNeo-SW, were constructed successfully. The expression level of survivin gene in SMMC-7721 cells was observed. After the treatment of RNAi, the expression of survivin gene in SMMC-7721 cells was almost absent, apoptosis index was increased by 15.6%, and the number of cells was decreased in G2/M phase and the cell growth was inhibited. CONCLUSION: RNAi can exert a knockdown of survivin gene expression in SMMC-7721 cells, and induce apoptosis and inhibit the growth of carcinoma cells.

Effect of Dexamethasone on Nitric Oxide Synthase and Caspase-3 Gene Expressions in Endotoxemia in Neonate Rat Brain

Objective To investigate the gene and protein expressions of three isoforms of nitric oxide synthase (NOS) and gene expression of Caspase-3, and effect of dexamethasone on them in neonatal rats with lipopolysaccharide (LPS)-induced endotoxemic brain damage. Methods Expressions of the three isoforms of NOS and caspase-3 mRNA in the brain were investigated by RT-PCR in postnatal 7-day wistar rats with acute endotoxemia by intraperitoneal administration of LPS. Regional distributions of NOSs were examined by immunohistochemical technique. Results nNOS and Caspase-3 mRNA were obviously detected. eNOS mRNA was faintly expressed, but iNOS mRNA was undetectable in the control rat brain. The expressions of NOS mRNA of three isoforms were weak 2 h after LPS (5 mg/mg) delivery, peaked at 6 h, and thereafter, reduced gradually up to 24 h. The expression intensity was in the order of nNOS> iNOS> eNOS. Widespread nNOS, scattered eNOS distribution and negative iNOS were identified in the control rat brain and all isoforms of NOS could be induced by LPS which reached the apex at 24 h in the order of nNOS> iNOS> eNOS as detected by immunostaining. Although Caspase-3 mRNA could be found in all groups, DNA fragmentation was only seen at 6 h and 24 h. The expressions of NOS and Caspase-3 mRNA were inhibited in the rat brain when dexamethasone was administrated. Conclusion LPS-induced NO production induces apoptosis of neurons through mechanism involving the Caspase-3 activation, which may play an important role in the pathogenesis of brain damage during endotoxemia, and neuro-protective effects of dexamethasone may be partially realized by inhibiting the expression of NOS mRNA.

Change in the body temperature of healthy term infant over the first 72 hours of life

Objective: To determine the range of body temperature in a group of healthy Chinese term neonates over the first 72 hours of life and to assess the influence of body weight, gestational age and route of delivery. Method: All 200 consecutive cases of neonates delivered at our hospital from March to August 2001 were included in this retrospective study. Temperatures were measured immediately after delivery, after 30 minutes, 1 hour, 2 hours, 8 hours and 15 hours and on the 2nd and 3rd day. Axillary temperatures ranging from 36.5 篊 to 37 篊 were regarded as normal. No cases of maternal fever or systemic infection of the newborns were discovered. All infants were discharged in good general condition. Results: The mean rectal temperature at birth was 37.19 篊. The lowest average temperature was reached at 1 hour after delivery (36.54 篊) with a significant difference between natural delivery (36.48 篊) and section (36.59 篊) (P<0.05). Temperature subsequently rose to 36.70 篊 at 8 hours and 36.78 篊 at 15 hours (P<0.05). Hypothermia was seen in 51.8% and hypothermia in 42.5% of the patients. On the 3rd day after delivery, 96% of all temperatures were in the normal range. A significant relation was found between hypothermia and both low birth weight (P<0.001) and low gestational age (P<0.05). Conclusion: The ref-erence range presently used did not include all physiological temperatures in the first 72 hours of life. Considering other factors, such as birth weight, route of delivery, gestational age and body temperature on the 2nd and 3rd day of life, may help to correctly assess the significance of temperatures beyond the reference range.

Expression of intestinal trefoil factor, proliferating cell nuclear antigen and histological changes in intestine of rats after intrauterine asphyxia

AIM:To study the expressions of intestinal trefoil factor (ITF) and proliferating cell nuclear antigen (PCNA) and histologic changes in intestine, to investigate the relationship between ITF and intestinal damage and repair after intrauterine hypoxia so as to understand the mechanism of intestinal injury and to find a new way to prevent and treat gastrointestinal diseases. METHODS: Wistar rats, pregnant for 21 d, were used to establish animal models of intrauterine asphyxia by clamping one side of vessels supplying blood to uterus for 20 min, another side was regarded as sham operation group. Intestinal tissues were taken away at 0, 24, 48 and 72 h after birth and stored in different styles. ITF mRNA was detected by RT-PCR. PCNA expression was measured by immunohistochemistry. Intestinal tissues were studied histologically by HE staining in order to observe the areas and degree of injury and to value the intestinal mucosa injury index (IMDI). RESULTS: ITF mRNA appeared in full-term rats and increased with age. After ischemia, ITF mRNA was decreased to the minimum (0.59?.032) 24 h after birth, then began to increase higher after 72 h than it was in the control group (P<0.01). PCNA positive staining located in goblet cell nuclei. The PCNA level had a remarkable decline (53.29±1.97) 48 h after ischemia. Structure changes were obvious in 48-h group, IMDI (3.40±0.16) was significantly increased. Correlation analyses showed that IMDI had a negative correlation with ITF mRNA and PCNA (r= -0.543, P<0.05; r= -0.794, P<0.01, respectively). CONCLUSION: Intrauterine ischemia can result in an early decrease of ITF mRNA expression. ITF and PCNA may play an important role in the damage and repair of intestinal mucosa.

Immune Intervention Effects on the Induction of Experimental Autoimmune Thyroiditis

To explore im mune intervention effects of the combined use of cycloporin A (Cs A ) and 1,2 5 - dihydroxyvitam in D3[1,2 5 (OH ) 2 D3]at low doses on experimental autoim mune thyroiditis (EAT) ,porcine thyroglobulin (p TG) was injected into a CBA mouse at the dose of 10 0μg on day 0 and day14 to establish the model of EAT.The immune prevention group from day0 to day2 8, and treatm ent group from day10 to day38were daily adm inistered Cs A(10 mg/ kg) intragastri- cally and/ or1,2 5 (OH) 2 D3(0 .2 μg/ kg) ip.After im munized by p TG,the m ice were sacrificed on day2 8and day38to examine their thyroid gland pathologically,and to check the levels of serum porcine thyroglobulin antibodies (p TGAb) ,porcine thyromicrosom al antibodies (p TMAb) .The incidences of EAT in the immune prevention group and treatment group,with administration of low dose of Cs A and 1,2 5 (OH) 2 D3,were decreased respectively by 4 4 .4 4 % and 37.5 0 % .Those of severe disease in the two groups were decreased respectively by71.4 3% and6 0 .32 % .The levels of serum p TGAb and p TMAb in the imm une prevention group were lower than those of the positive control group.It was concluded thatcombined use of Cs A and1,2 5 (OH) 2 D3at low doses could effectively prevent EAT with a synergic effect.

Effect of Retinoic Acid on Lung Injury in Hyperoxia-Exposed Newborn Rats

To investigate whether treatment with retinoic acid (RA) could improve level of lung alveolarization and influence lung collagen in newborn rats exposed to hyperoxia, newborn Sprague-Dawley rats aged 2 days were randomly assigned to 8 groups:(1) air, (2) O 2, (3) air+NS, (4) O 2+NS, (5) air+dex, (6) O 2+dex, (7) air+RA and (8) O 2+RA. Group 2, 4 6 and 8 were kept in chambers containing 85 % oxygen, the values were checked 3 times a day. The other 4 groups were exposed to room air. Level of alveolarization and lung collagen were analyzed at age of 14 or 21 days through radial alveolar counts, alveolar airspace measurements, type Ⅰ, Ⅲ collagen immunohistochemical methods (SP method) and image processing system. Transforming growth factor-β receptors and procollagen mRNA accumulation were examined at age of 14 days through immunohistochemical methods and in situ hybridization. Our results showed that radial alveolar counts were increased and distal airspace was enlarged in group 8. TypeⅠcollagen was markedly increased, and transforming growth factor-β receptors and procollagen mRNA were decreased by retinoic acid in bronchial epithelial cells, alveolar epithelial cells and alveolar intersitium. It is concluded that retinoic acid can partially reverse lung development arrest during exposure to hyperoxia by increasing lung collagen.

Shp-2／NF-κB Pathway Mediates the Inhibition of Lipoxin A4 onIL-1β-induced Synthesis of IL-6 in Glomerular Mesangial Cells

Objective: To examine whether lipoxin A 4 (LXA 4) has an antagonistic effect on IL-1β-induced synthesis of IL-6 in glomerular mesangial cells, and to explore the molecular mechanisms of signal pathway in LXA 4 actions. Methods: The glomerular mesangial cells of rat were cultured and treated with IL-1β, with or without preincubation with LXA 4 at different concentrations. The amount of IL-6 in the supernatant of cells was analyzed by enzyme-linked immunosorbent assay(ELISA). The expressions of mRNA of IL-6 were determined by RT-PCR. The expressions of Src homology 2(SH 2) containing protein-tyrosine phosphatase 2(Shp-2) were assessed by immunoprecipitation and immunoblotting. Activities of DNA-binding of nuclear factor-kappa B(NF-κB) were measured by electrophoretic mobility shift assay(EMSA). Results: IL-1β-stimulated secretion of protein and expression of mRNA of IL-6 in mesangial cells were inhibited by LXA 4 in a dose-dependent manner. LXA 4 antagonizes the phosphorylation of Shp-2 and activities of NF-κB induced by IL-1β. Conclusion: LXA 4 antagonists IL-1β-induced synthesis of IL-6 in glomerular mesangial cells through the mechanism of Shp-2/NF-κB pathway-dependent signal transduction.

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.

EFFECTS OF CHINA-MADE RECOMBINANT HUMAN GROWTH HORMONE ON THE TREATMENT OF GROWTH HORMONE DEFICIENCY

To evaluate the therapeutic effect of China-made recombinant human growth hormone (r-hGH) in children with growth hormone deficiency (GHD) and to investigate the utilities of various biochemical parameters in GHD diagnosis and treatment. Methods Our study comprises of 30 normal children and 71 GHD children treated with China-made r-hGH substitution therapy 0.1 IU·kg-1·d-1 for 6 months. Serum insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein 3 (IGFBP-3), bone turnover markers (Ost, ICTP), and anti-growth hormone antibody (GHAb) were detected before and after r-hGH treatment. Results After the first 3 and 6 months of treatment, growth velocities of GHD children were significantly increased (13.1 ± 3.7 and 12.6 ± 3.6 cm/year) compared with pretreatment values (2.9 ± 0.8 cm/year, P < 0.01). GHD Children had obviously reduced serum levels of IGF-1, IGFBP-3, and bone turnover markers (Ost, ICTP) compared with normal controls (P < 0.01), and these biochemical parameters improved significantly after treatment (P < 0.01). Growth hormone antibodies were positive in 17 of 45 cases after treatment by binding capacity detection. The binding percentage of growth hormone an-tibody which was increased more than 30% after the treatment showed a negative correlation with growth velocity (P < 0.01). Conclusions (1) The growth stimulating effect and safety were confirmed in using China-made r-hGH in the treatment of GHD children for 6 months. (2) The measurements of serum IGF-1 and IGFBP-3 may serve as useful parameters in the diagnosis of GHD. (3) Serum Ost and ICTP are useful laboratory criteria for evaluating the effect of r-hGH therapy in the early stage. (4) It is necessary to monitor serum levels of GHAb during r-hGH therapy.

Structure prediction and activity analysis of human heme oxygenase-1 and its mutant

AIM: To predict wild human heme oxygenase-1 (whHO-1) and hHO-1 His25Ala mutant (△hHO-1) structures, to done and express them and analyze their activities.METHODS: Swiss-PdbViewer and Antheprot 5.0 were used for the prediction of structure diversity and physicalchemical changes between wild and mutant hHO-1, hHO-1 His25Ala mutant cDNA was constructed by site-directed mutagenesis in two plasmids of E.COli DH5α Expression products were purified by ammonium sulphate precipitation and Q-Sepharose Fast Flow column chromatography, and their activities were measured,RESULTS: rHO-1 had the structure of a helical fold with the heme sandwiched between heme-heme oxygenase-1 helices. Bond angle, dihedral angle and chemical bond in the active pocket changed after Ala25 was replaced by His25, but Ala25 was still contacting the surface and the electrostatic potential of the active pocket was negative. The mutated enzyme kept binding activity to heme. Two vectors pBHO-1 and pBHO-1(M) were constructed and expressed. Ammonium sulphate precipitation and column chromatography yielded 3.6-fold and 30-fold higher purities of whHO-1, respectively. The activity of △hHO-1 was reduced 91.21% after mutation comparedwith whHO-1.CONCLUSION: Proximal His25 ligand is crucial for normal hHO-1 catalytic activity. △hHO-1 is deactivated by mutation but keeps the same binding site as whHO-1. △hHO-1 might be a potential inhibitor of whHO-1 for preventing neonatal hyperbilirubinemia.

Detection of T lymphocyte subsets of children with Helicobacter pylori infection

AIM: To study the transformation of T lymphocyte subsets in children with Heliobacter pylori (H pylori) infection. METHODS: The H pylori infection status were determined by a combination of ELISA and Western blot (immunoblot) technique in 98 children and T lymphocyte subsets in peripheral blood were determined by flow cytometrical analysis.RESULTS: There were 75 children positive with H pylori infection and 23 negative in 98 children. Comparing the proportion of peripheral blood T lymphocytic subsets in children with H pylori infection and without H pylori infection, it was found that a higher proportion of CD4 T-cells in infected children (39.02±7.71 vs34.25±10.73,t = 2.246,P＜0.05) and higher value of CD4 to CD8 T-cells ratio (1.51±0.52 vs 1.25, t= 2.104,P＜0.05) were present, but there were not significant differences in CD3 T-cells and CD8 T-cells (73.11±10.02 vs69.49±17.08, 27.22±6.07vs 28.27±8.67, P＞0.05).CONCLUSION: Th1 cell-mediated immune responses may be induced by H pylori infection in children.

肺炎支原体感染

肺炎支原体是儿童和成人社区获得性呼吸道感染的常见病因。尽管其被认为是成人社区获得性肺炎最常见的非典型病原体，但因人群和诊断方法不同，不同研究之间肺炎支原体感染的发病率差异很大。近期研究显示，成人肺炎中，支原体肺炎发病率从1．9％到30％不等。肺炎支原体也是导致社团呼吸道感染爆发流行的常见原因，肺炎支原体感染的诊断受其缺乏标准、快速、特异性诊断手段的影响。联邦训练局对一次肺炎支原体感染爆发流行的调查结果是上述问题的很好例子。确诊需聚合酶链反应(PcR)和血清学二者均阳性；许多肺炎支原体感染早期并不出现IgM抗体。在本次文献复习期间，又有几篇评价各种血清学方法和PcR检测的论文发表。对这些检验方法的评价受到缺乏标准比较方法的妨碍。在体外，肺炎支原体对大环内酯类、四环素类和喹诺酮类抗微生物药均敏感；但有关这些药物的抗微生物疗效的数据很少。同期发表了几项肺炎治疗的研究文章，根据血清学阳性结果作出肺炎支原体感染的诊断并应用上述药物，每项研究采用的方法和判断标准不同，因此无法评价上述各种药物的抗微生物学疗效。美国感染病学会在最近修订的成人社区获得性肺炎治疗的实践指南中表示，因缺乏快速可靠的诊断试验，肺炎支原体感染多为经验性治疗。

α-Tocopherol Concentrations in Human Milk from Mothers of Preterm and Full-term Infants in China

a-Tocopherol content in breast milk of 28 mothers who delivered preterm infants (preterm milk) and 43 mothers who delivered full-term infants (term milk) were measured, a-Tocopherol concentration in preterm milk did not differ significantly from that of term milk in the first 12 days of lactation (P>0.05). There is a higher a-tocopherol concentration in the early colostrum, however, it decreases with the lactational days significantly. The investigation suggests that early breast-feeding would be beneficial to the improvement of vitamin E intake in neonates during the early life.

Risk Factors for Cardiovascular Disease in Obese and Normal School Children: Association of Insulin with Other Cardiovascular Risk Factors

Objective To investigate the level of insulin and other known cardiovascular risk factors in school children and their association with obesity.Design Cross-sectional study.Subjects 123 normal and 116 obese school children categorized by weight-for-height,mean age. 10.2 and 10.5 years old respectively.Measurements Family histories of diseases by questionnaires; blood pressure (BP) and waist and hip circumferences by measurements; fasting blood for glucose, insulin, total cholesterol, HDL-cholesterol and triglycerides.Results The numbers of boys in the normal and obese group were 65 and 58, those of the girls, 53 and 63 respectively. The obese group was more likely to have family histories of obesity, high blood pressure and diabetes; had significantly greater waist-hip-ratio (WHR), higher systolic and diastolic BP, lower HDL-cholesterol, higher triglyceride and fasting insulin levels (with fasting blood sugar in the normal range) than the normal weight group. In a crude analysis, insulin levels were positiveIy correlated with obesity, systolic BP, WHR, age and triglycerides and negatively associated with male gender and HDLcholesterol. After adjustment using multiple regression, only obesity status, age, gender and triglycerides still remained significantly associated with insulin level. Limitation of utilizing family disease history report for identification of children at risk was discussed.Conclusion These findings suggest that risk to coronary heart disease and hypertension through insulin resistance already operates in school-aged children

Molecular Mechanisms of Cell-cell Recognition

Cell-cell recognition is the key for multicellular organisms to survive. This recognition critically depends on protein-protein interactions from opposing cell surfaces. Recent structural investigations reveal unique features of these cell surface receptors and how they interact. These interactions are specific, but usually relatively weak, with more hydrophilic forces involved in binding. The receptors appear to have specialized ways to present their key interacting elements for ligand-binding from the cell surface. Cell-cell contacts are multivalent. A large group of cell surface molecules are engaged in interactions. Characteristic weak interactions make possible for each individual molecule pair within the group to constantly associate-dissociate-reassociate, such that the cell-cell recognition becomes a dynamic process. The immunological synapse is a good example for immune receptors to be orchestrated in performing immunological function in a collective fashion.

Reevaluation of the Efficacy of Intravenous Gammaglobulin in the Prevention and Treatment of Coronary Artery Lesion in Kawasaki Disease

In order to objectively evaluate the efficacy of intravenous gammaglobulin (IVIG) in the prevention and treatment of coronary artery lesion (CAL) in Kawasaki disease (KD) and the related factors influencing the IVIG efficacy, 314 children with KD were reviewed retrospectively and comparatively and were divided into IVIG plus aspirin group and ASA group. The occurrence and restoration of CAL in these two groups as well as many laboratory and clinical indexes including average hospital stay (days), total fever duration, defervescence time, platelet count, erythrocyte sedimentation rate, C reactive protein etc. were observed. The incidence of CAL was 39.5 % in the children with KD. In the IVIG+ASA group, the incidence of CAL was 34.3 % and 56.0 % in ASA group respectively (P<0.001). The incidence of CAL was reduced in the group in which 2.0 g/kg or 1.0 g/kg IVIG was administered as compared with the group in which IVIG was administered at a dose ≤0.6 g/kg or ≥3.0 g/kg (P<0.05). CAL occurred less frequently when IVIG was administered at 3-10 days of the course than that when IVIG was administered ≤3 days or >10 days (P<0.05). About 13.4 % of the CAL treated with IVIG was not recovered at the 12 th month of the course, mostly in the groups in which only ASA was administered and IVIG treatment was started 10 days later. The hospital stay (days), defervescence time, total fever duration, platelet count, erythrocyte sedimentation rate and C reactive protein were significantly reduced in IVIG+ASA group as compared with those in the ASA group (P<0.05). IVIG treatment can remarkably shorten the course of patients with KD and decrease the incidence of CAL, but the efficacy of IVIG in the prevention and treatment of KD disease is not as expected by people, therefore, reevaluation of the practical efficacy of IVIG is required.