Expression of nucleotide-binding oligomerization domain-like receptor protein 3 inflammatory corpuscles in epileptic model and the effect of melatonin on its expression / 中华实用儿科临床杂志

Objective To observe the expression of nucleotide-binding oligomerization domain-like recep-tor protein 3(NLRP3)inflammasome in epilepsy model,and to explore the neuroprotective effect of melatonin. Methods SD rats aged 21-30 d were randomly divided into the control group(48 rats),the epilepsy group(48 rats)and the melatonin group(48 rats),and each group was subdivided into 4 subgroups according to the time points of 24 h,48 h, 72 h,and 7 d,with 12 SD rats in each subgroup. Fluorescence quantitative polymerase chain reaction and immunohisto-chemical technique were used to analyze the expressions of NLRP3,Caspase-1 and interleukin(IL)-1β in hippocam-pus areas of rats at different points of time after seizures were induced,and their behavior changes were observed. Results The number of NLRP3-positive cells in the epileptic group increased,and reached the peak at 72 h. At 24 h,48 h,72 h,7 d,the number of NLRP3-positive cells in the epilepsy group(14. 20 ± 1. 64,23. 60 ± 1. 14,31. 20 ± 1. 30,25. 40 ± 2. 07)was significantly increased compared with those of the melatonin group(10. 60 ± 0. 89,17. 80 ± 1. 48,24. 00 ± 0. 71,20. 20 ± 1. 92)and the control group(2. 60 ± 0. 89,2. 40 ± 1. 14,2. 40 ± 1. 14,2. 40 ± 0. 55),and the differences were significant(F＝122. 977,375. 125,962. 743,262. 916,all P＜0. 05). The NLRP3 mRNA relative expressions in the epilepsy group (2. 57 ± 0. 12,3. 34 ± 0. 10,4. 84 ± 0. 19,3. 55 ± 0. 13)were significantly increased compared with those of the melatonin group (2. 03 ± 0. 08,2. 71 ± 0. 08,4. 03 ± 0. 14,2. 48 ± 0. 18)and the control group(1. 07 ± 0. 13,1. 08 ± 0. 15,1. 08 ± 0. 23,1. 07 ± 0. 18),and the differences were significant (F ＝422. 386, 1 154. 957,1 132. 112,512. 149,all P ＜0. 05);the Caspase -1 mRNA relative expressions in the epilepsy group (2. 47 ± 0. 07,3. 05 ± 0. 15,4. 39 ± 0. 18,3. 14 ± 0. 11)were significantly increased compared with those of melatonin group(1. 85 ± 0. 07,2. 49 ± 0. 08,3. 60 ± 0. 12,2. 15 ± 0. 12)and the control group (0. 98 ± 0. 25,0. 99 ± 0. 15,0. 98 ± 0. 23,0. 99 ± 0. 18),and the differences were significant(F ＝620. 099,580. 796,1 125. 225,645. 082,all P ＜0. 05);the IL-1β mRNA relative expressions in epilepsy group (2. 32 ± 0. 15,2. 90 ± 0. 18,4. 18 ± 0. 16,2. 74 ± 0. 07)were significantly increased compared with those of the melatonin group (1. 78 ± 0. 09,2. 35 ± 0. 11,3. 24 ± 0. 13,1. 78 ± 0. 16)and the control group(0. 97 ± 0. 13,0. 99 ± 0. 15,0. 97 ± 0. 23,0. 97 ± 0. 18),and the differences were significant(F＝267. 952,398. 767,1 140. 384,438. 962,all P ＜0. 05). Conclusions The NLRP3 inflamma-somes are activated in rat hippocampus with epilepsy induced by lithium-pilocarpine. NLRP3 inflammasome mediated inflammatory response probably involved in the pathogenesis of epilepsy. The melatonin may play a neuroprotective role by inhibiting expression of NLRP3 inflammasome.

The Study on Maternal Plasma Placental Growth Factor Level and the Perinatal Outcome in Early-Onset Pre-Eclampsia Patients / 实用妇产科杂志

Objective:To investigate the relationship between plasma placental growth factor(PIGF) levels and perinatal outcomes in patients with early onset preeclampsia(EOPE).Methods:125 patients with EOPE undergoing cesarean section were enrolled in this study.Enzyme-linked immunosorbent assay(ELISA) was used to detect the level of maternal plasma PIGF.According to the plasma PIGF level,≤ 12 pg/ml was divided into PlGF≤12 pg/ml group(n =64) and PIGF ＞12 pg/ml was in PIGF ＞12 pg/ml group(n =61).The blood gas analysis and the level of lactic acid(LAC) were measured within 1 hour after fetal birth.The routine and biochemical indexes of two groups and their relation with perinatal outcomes were compared.Results:①In PIGF ≤ 12 pg/ml EOPE group,the Hb,HCT,24h urine protein level,BUN and Cr levels were higher than those in PIGF ＞ 12 pg/ml group (P ＜ 0.05).While the ALB level was lower than that of PIG F ＞ 12 pg/ml group (P ＜ 0.05).②The gestational age of PIGF≤12 pg/ml group was less than that of PIGF ＞ 12 pg/ml group,but the proportion of oligohydramnios,meconium stained amniotic fluid and less than gestational age infants(SGA) were all higher than those of PIGF ＞ 12 pg/ml group(P＜0.05).③In the PIGF≤12 pg/ml group,neonatal birth weight(BW),1 minute Apgar score,pH,serum Ca,BE were all lower than those in PIGF ＞12 pg/ml group.But the level of LAC and the NICU hospital stay were all higher than those in PIGF ＞ 12 pg/ml group (P ＜ 0.05).Conclusions:In EOPE patients,plasma PIGF level has certain clinical value in predicting perinatal outcome.If PIGF is ≤ 12 pg/ml,the maternal renal dysfunction may be more obviously,and there is higher incidence of oligohydramnios,meconium stained fluid,SGA and neonatal asphyxia,and the newborn is more susceptible to metabolic imbalance and acid-base disorders.Mother and child care should be strengthened.

Comparison of efficacy of adrenocorticotrophic hormone and methylprednisolone on rat models of infantile spasm / 中华实用儿科临床杂志

Objective To compare the efficacy of adrenocorticotrophic hormone (ACTH) and methylprednisolone on the rat models of infantile spasms (IS).Methods The SD rats on postnatal 10 day (P10) were divided into blank group (n =18),control group (n =18) and model group (n =110) according to the random number table method.The rats of model group were prepared by adopting prenatal stress exposure and N-methyl-D aspartate (NMDA) injection.In the model group,after inducing epileptic seizures,the rats were divided into different groups (18 rats in each group) according to the random number table method as following:model group Ⅰ (subcutaneous injection ofACTH,50 IU/kg,at P10:14:00,21:00;P11,P12:7:00,14:00,21:00;P13:7:00),model group Ⅱ (subcutaneous injection of 9 g/L saline),model group Ⅲ (intraperitoneal injection of methylprednisolone,60 mg/kg,at P11,P12,P13:9:00,once per day),model group Ⅳ (intraperitoneal injection of 9 g/L saline) and model group Ⅴ (positive control group,with no drug or saline injection).Three days later,epilepsy was induced again,and the rats of model group were intraperitoneally injected with NMDA (12 mg/kg) at P13 (10:00).The rats of control group were injected intraperitoneally with same volume of 9 g/L saline,but the rats of blank group were not treated.Behaviors of rats with epilepsy seizures were observed and epilepsy scores were given.The expression of corticotropin-releasing hormone (CRH) in the hypothalamus of each group was detected by using immunohistochemistry and fluorescence quantitative polymerase chain reaction.The learning and memorizing capacity of the rats were measured by Y-maze experiment.Results There was no death in the model group after the onset of seizure.In the model group Ⅰ,13 cases were attacked(72.22％),and 14 cases were attacked in the model group Ⅲ (78.78％).The level of attack was decreased.The buckling state was not observed in model group and Ⅲ,but the latency period of epilepsy was prolonged and the epilepsy scores were significantly decreased.There were no significant differences of onset latency [(2 369.38 ± 628.70) s vs.(1 922.93 ± 462.36) s] and epilepsy score [(2.15 ± 1.14) scores vs.(2.07 ± 0.83) scores] between the 2 groups (all P ＞ 0.005).The rats of model group Ⅱ,Ⅳ,Ⅴ were all attacked completely and presented buckling state.There was no onset or death in blank group and control group.The number of CRH positive cells and CRH mRNA relative expression of each model group were higher than those in the blank group and control group.The number of CRH positive cells and CRH mRNA expression of model group Ⅰ and Ⅲ were lower than those in model group Ⅱ,Ⅳand Ⅴ,and the differences were significant (all P ＜ 0.002 4).There was no significant difference in the number of CRH-positive cells(39.12 ± 5.98 vs.41.48 ± 7.61) and CRH mRNA relative expression (1.92 ± 0.16 vs.2.06 ± 0.39) between model group Ⅰ and Ⅲ (all P ＞ 0.002 4).No significant difference was found between blank group and control group,or among model group Ⅱ,Ⅳ and Ⅴ (all P ＞ 0.002 4).There were no significant differences in the learning capacity among all groups (F =2.196,P ＞ 0.002 4).The correct response rate after 24 hours of the model group was lower than the blank group and control group,and ACTH and methylprednisolone pretreatment did not influence the memorizing capacity (P ＞ 0.002 4).Conclusion The effect of pretreatment of ACTH is similar to that of methylprednisolone in the rat model of IS.

Identification of types of GABAA receptors in neurons in brain tissues at target of anesthetic action of isoflurane in mice / 中华麻醉学杂志

Objective To identify the types of γ?aminobutyric acid type A ( GABAA ) receptors in neurons in brain tissues at the target of anesthetic action of isoflurane in mice. Methods Two mouse strains were developed that were either sensitive or resistant to isoflurane. One hundred isoflurane?sensitive ICR∕CD?1 mice ( 50 males, 50 females) and 100 isoflurane?resistant ICR∕CD?1 mice ( 50 males, 50 fe?males) , aged 65-70 days, were used in this study. Brain tissues were obtained, and total RNA was ex?tracted and then reverse transcribed to cDNA using AMV reverse transcriptase. Polymerase chain reaction was used to detect the cDNA sequences. Chi?square analysis was used to compare the cDNA sequence of each GABAA receptor subunit between two strains. Results The cDNA sequence of GABAA receptor sub?units α1?6 , β2,3 andγ1?3 in isoflurane?sensitive strain was completely consistent with that in isoflurane?resist?ant strain. A single nucleotide polymorphism at the nucleotide position 462 ( C∕G) in the β1 sequence was found. The allele C frequencies were 11.0% and 87.0% in isoflurane?sensitive strain and isoflurane?resistant strain, respectively. Compared with isoflurane?sensitive strain, the allele C frequency in cDNA sequences of β1 subunit was significantly increased in isoflurane?resistant strain ( P<0.01) . Conclusion β1 subunit?containing GABAA receptor in neurons in brain tissues is the target of anesthetic action of isoflurane in mice.

The effect of different dosages of low molecular weight heparin on acute pulmonary embolism and inhibition of pulmonary intimal hyperplasiain immature rats / 临床儿科杂志

Objectives To investigate the effect of different dosages of low molecular weight heparin on acute pulmonary embolism and inhibition of pulmonary intimal hyperplasia in immature rats. Methods 90 male immature SD rats were randomly divided into ifve groups: sham group, pulmonary embolism group, low-low molecular heparin group (L-LMH), medium-low molecular heparin group (M-LMH) and high-low molecular heparin group (H-LMH). The model of acute pulmonary embolism was established through jugular vein injection with gel-foam solution. The rates in the L-LMH, M-LMH, H-LMH groups were treated with low molecular weight heparin by subcutaneous injection after surgery with a dosage of 0 . 005 ml/kg, 0 . 01 ml/kg, 0 . 02 ml/kg, twice a day. Animals in the control group were given saline injection. Arterial blood gas, pulmonary artery pressure (mPAP), right ventricular pressure (RVP), wall area/tube area, wall thickness/tube diameter, and the expression of PDGF-B and MCP-1 at gene and protein levels in lung tissue were detected on the 7 th ( 7 d), 14 th ( 14 d) and 28 th ( 28 d) after opration. Results There were signiifcant differences of PaO 2 among 5 groups on 7 d, 14 d and 28 d. PaO 2 in group M-LMH ( 105 . 1 ± 4 . 6 mm Hg) were signiifcantly higher than that of embolization group, L-LMH, but not H-LMH group at 28 d. mPAP of M-LMH group was lower than that in the other three intervention groups, but showed no signiifcant difference compared with sham group (P?>0 . 05 ). There were signiifcant differences of RVP on 7 d and 14 d. PDGF-B, MCP-1 of M-LMH group were signiifcantly lower compared with the other three intervention groups (P?0 . 05 ). Wall area/tube area, wall thickness/tube diameter scores of M-LMH group had no signiifcance differences compared with sham group on 28 d (P?>?0 . 05 ). Conclusion Medium dose of low molecular weight heparin could ameliorate the acute pulmonary embolism and inhibit the proliferation of pulmonary arteries in rats.

Analysis of clinical characteristics and perinatal outcomes in pre-pregnancy obesity patients with early-onset preeclampsia / 天津医药

Objective To explore clinical characteristics and the perinatal effects of multiple systems in pre-pregnancy obesity patients with early-onset preeclampsia. Methods A total of 111 cases with early-onset preeclampsia in our Inpatient Obstetric Department were reviewed in this study. According to pre-pregnancy body mass index (BMI), patients were divided into normal before pregnancy group (BMI<28 kg/m2, n=56) and obese before pregnancy group (BMI≥28 kg/m2, n=55) . The body weight, BMI, weight gain during pregnancy, risk factors, clinical features, and clinical features of neonatal index were compared between two groups. The correlation between maternal and neonatal indexes and glucose and lipid indexes were compared. Results Obese before pregnancy was more prone to blood concentration and disorders of lipid metabolism. The serum levels of lipid, glycosylated hemoglobin (HbA1c), hematocrit, platelet and fibrinogen were significantly higher in obese before pregnancy group than than those of normal group. The induced neonatal acidosis and other organ damage, newborn Apgar score, pH, glucose were significantly lower in obese before pregnancy group than those of normal group. The residual alkali (BE), lactic acid (LAC), creatine kinase (CK) and creatine kinase (CKMB) were higher in obese before pregnancy group than those of normal before pregnancy group. Correlation analysis showed that maternal low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), three acyl glycerol (TG) and HbA 1c were negatively correlated with neonatal Apgar score and pH, and which were positively correlated with neonatal LAC and CKMB. Both differences were significant (P<0.05 or P<0.01). Conclusion The dyslipidaemia in early onset preeclampsia in obesity patients before pregnancy is closely related with maternal and perinatal outcomes.

Changes of IL-8 and ICAM-1 in early stage of cerebral white matter damage and their correlations with prognosis in premature infants / 临床儿科杂志

Objective To explore the IL-8 and ICAM-1 in premature infants with cerebral white matter damage (CWMD) and their correlations with prognosis. Methods One hundred and two cases of CWMD were selected from March 2009 to June 2012 as experimental group and 42 cases of normal preterm children were selected as control group. Serum IL-8 and ICAM-1 levels were measured during 48-72 h after birth. Motor development index (psychomotor developmental index, PDI) and mental development index (mental development index, MDI) were evaluated by Bayley scale. The correlations of IL-8 and ICAM-1 levels with prognosis were analyzed. Results Serum IL-8 and ICAM-1 levels in the experimental group were signiifcantly higher than those in the control group (P<0.05). MDI and PDI scores in the experimental group were signiifcantly lower than those in the control group (P<0.05). There were negative correlations of serum IL-8 and ICAM-1 levels with MDI (r=-0.64, P<0.05;r=-0.66, P<0.05)+and PDI (r=-0.70, P<0.05;r=-0.71, P<0.05). Conclusions Serum IL-8 and ICAM-1 levels in infants with CWMD are signiifcantly increased, and are negatively correlated with MDI and PDI scores.

Immunomodulatory effects of 1,25 dihydroxyvitamin D3 in the treatment of experimental colitis / 中华消化杂志

Objective To explore the immunomodulatory effects of 1,25-dihydroxyvitamin D3 (1,25 (OH)2D3) in the treatment of experimental colitis induced by dextran sulfate sodium (DSS) in mice.Methods According to random number table,thirty BALB/c mice were randomly divided into control group,model group,low dose,moderate dose,and high dose intervention group.Mice of model group,low dose,moderate dose and high dose intervention group drank 5％ DSS solution for seven days to create colitis model.On the 1st,3rd,5th,7th day,the mice of low dose,moderate dose and high dose intervention group were intraperitoneal injected with low,moderate,and high dose of 1,25(OH)2D3 (50,100 and 200 ng/each mouse,respectively).Mice of control group and model group were intraperitoneal injected with sterile soybean oil as control.The observed indicators included disease activity index (DAI) and colonic histopathological score (HPS).On the 8th day,all mice were sacrificed.The expression of interferon (IFN)-γ,interleukin (IL-17) and IL-21 in mice colon tissues and spleens at mRNA and protein level were measured by reverse transcription-polymerose chain reaction (RT-PCR) and flow cytometry,respectively.The data were analyzed by one way ANOVA.LSD-test or Tamhane test were performed for comparison in groups.Results Compared with the control group,the DAI and colitis HPS of mice in the model group significantly increased (0.33±0.52 vs 7.33±1.03,0.17±0.41 vs 12.00±0.63).Compared with the model group,the DAI and colonic HPS of intervention groups treated with 1,25 (OH)2 D3 declined with varying degrees (2.83 ± 0.40,2.83±0.75,2.33±0.52 and 10.83±0.98,7.50±0.84,6.67±0.52,LSD-t=0.39 and 0.41,all P＜0.01).The expression of IFN-γ,IL-17 and IL-21 of the model group were significantly higher than those of the control group.The expressions of IFN-γ,IL-17 and IL-21 of intervention group were signifiantly lower than that of the model group (mRNA:LSD-t =0.12,0.13,0.09; protein:F =20.61,22.46,4.80,all P＜0.01).Conclusion 1,25 (OH)2D3 might have a direct role on T-cell phenotype,down-regulate effective cytokines IFN-γ,IL-17 and IL-21 and then play an interventional role.

DNA-EGS1386 in cells induced RNase P inhibits the expression of human cytomegalovirus UL49 gene / 生物工程学报

External Guide Sequences (EGSs) represents a novel nucleic acid based gene interference approach to modulate gene expression. They are oligonucleotides that consist of a sequence complementary to a target mRNA and recruit intracellular RNase P for specific degradation of the target RNA. DNA-based EGS1386 with a size of 12 nt was chemically synthesized to target the mRNA coding for the UL49 gene of human cytomegalovirus (HCMV). The DNA-based EGS1386 molecule efficiently directed human RNase P to cleave the target mRNA sequence in vitro. A reduction of more than 50% in the levels of UL49 expression was observed in human cells treated with the DNA-based EGS1386 targeted UL49 assayed by fluorescent quantization PCR and Western blotting. This results showed that the DNA-EGS1386 can effectively guide the RNase P cut the target mRNA. Therefore, DNA-EGS can develop into a new gene silencing technology and potential of the anti-viral reagents.

Adenoviral-mediated somatostatin receptor subtype 2 gene transfer inhibits pancreatic cancer xenografts / 中国实用内科杂志

Objective To construct recombinant adenovirus carrying the gene of human somatostatin receptor subtype 2(SSTR2),to investigate its effect on growth of human pancreatic cancer xenografts in nude mice,and to further elucidate the underlying mechanisms.Methods The recombinant adenovirus coding for human SSTR2(Ad-SSTR2)and reporter gene LacZ(Ad-LacZ)were generated by site-specific recombination from Jun 1,2004 to Dec 10,2005.Human pancreatic cancer cell line was implanted subcutaneously into nude mice;normal saline(control group),Ad-LacZ(reporter gene control group)and Ad-SSTR2(experimental group)were injected into pancreatic cancer xenografts,respectively.The tumor volume and weight were measured.RTPCR and Western blot were used to determine the expression of SSTR2 after pancreatic cancer xenografts were infected with Ad-SSTR2.The expression level of ERK2 and ras proteins were assessed by Western blot.Results The virus titer of Ad-SSTR2 and Ad-LacZ was 6.0?1012pfu/L and 6.5?1012pfu/L,respectively.SSTR2 mRNA and protein were detected after Ad-SSTR2 infected pancreatic cancer xenografts.Growth of pancreatic cancer was significantly inhibited in the experimental group as compared with the control group.The growth inhibitory rate was 48.2%(P

Construction and expression of recombinant adenovirus vector encoding human somatostatin receptor type 2 / 中国病理生理杂志

AIM: To construct recombinant adenovirus vector carrying the gene of human somatostatin receptor type 2(SSTR2) for gene therapy of pancreatic carcinoma.METHODS: SSTR2 cDNA was inserted into adenovirus shuttle plasmid pDC316,named pDC316-SSTR2.pDC316-SSTR2 was cotransfected with rescue plasmid pBHGlox(delta) E1,3Cre into 293 cells by liposome reagent.Ad-SSTR2 was generated by site-specific recombination and confirmed by PCR.Ad-SSTR2 was propagated in 293 cells and purified.The titer of viral stock was determined by end-point dilution assay.Western blotting was used to determine the expression of SSTR2 protein after human pancreatic carcinoma cell capan-2 was infected with recombinant adenovirus.RESULTS: pDC316-SSTR2 was successfully constructed.Recombinant adenovirus Ad-SSTR2 was acquired by pDC316-SSTR2 and pBHGlox(delta) E1,3Cre cotransfected into 293 cells.Ad-SSTR2 was characterized by PCR.The virus titer was 6.0?10~(12) pfu/L.SSTR2 protein was detected after adenovirus infected capan-2 48 h with Western blotting.CONCLUSION: The recombinant adenovirus vector encoding human SSTR2 is successfully constructed and correctly expressed in pancreatic carcinoma cells.This investigation provides the basis for study of gene therapy of pancreatic carcinoma.

Interrelation between the structure and the function of artificial ribozyme M_1GS / 中国病理生理杂志

AIM: To study the interrelation between the structure and the function of artificial ribozyme M_1GS. METHODS: Ribozyme M_1GS-T_7, which targeted the mRNA segment of HCMV UL54 gene, was constructed. The secondary structure of M_1GS was simulated under different temperatures (20 ℃, 37 ℃ or 55 ℃, at which the secondary structure of M_1GS was relatively stable) and the interrelation between the secondary structure and the cleavage activity of M_1GS was analyzed under different temperature in vitro. To investigate the interrelation between the structure and the function of ribozyme M_1GS further, mM_1GS-T_7 was designed, in which some mutation sites was added, according to the result of temperature change experiment and the simulated secondary structure, showing that were the same structures at 37 ℃ as that of M_1GS-T_7 at 55 ℃. RESULTS: In temperature change experiment, the cleavage activity of M_1GS-T_7 was highest at 55 ℃. The result of mutant experiment showed that the mutant type was more active than M_1GS-T_7 at 37 ℃. CONCLUSION: The cleavage activity of M_1GS, which has some certain secondary structure, is higher than others. There is some interrelation between the structure and the function of M_1GS.

A summary of the study on the homeobox gene Pem / 中国病理生理杂志

Pem gene is one of the homeobox gene. Unlike members of Hox gene family, Pem gene is unique located at the proximal end of the X chromosome and its expression has been detected in several immortalized and transformed cells, placenta, embryos and reproductive tissues. Numeral studies showed that its expression is controlled by hormone such as androgen. This review discussed the possible role of Pem in regulating genes involved in the differentiation and development of extraembryonic tissue, spermatogenesis and sperm maturation.