Analysis of clinical characteristics and genetic variants in two children with Limb-girdle muscular dystrophy autosomal recessive 9 FKRP-related / 中华医学遗传学杂志

OBJECTIVE@#To explore the correlation between clinical manifestations of Limb-girdle muscular dystrophy autosomal recessive 9 FKRP-related (R9 FKRP-related) and variants of the FKRP gene.@*METHODS@#Two children who had presented at the Children's Hospital of Nanjing Medical University respectively due to increased serum myocardial zymogram and hepatic dysfunction on September 30, 2018 and August 3, 2018 were selected as the study subjects. Clinical data of the children were collected. Both children were suspected for Duchenne or Becker muscular dystrophy for asymptomatic high creatine kinase (CK) levels. Peripheral blood samples of the children and their parents were collected for whole exome sequencing, and candidate variants were validated by Sanger sequencing.@*RESULTS@#Genetic testing revealed that both children have carried compound heterozygous variants of the FKRP gene. The c.545A>G and c.941C>T variants in child 1 have been reported previously, among which the c.545A>G is a hot spot mutation in the Chinese population. Child 2 has carried c.602T>C and c.961G>A variants, both of which were unreported previously.@*CONCLUSION@#Both children have met the diagnostic criteria for LGMD R9 FKRP-related. Carriers of the c.545A>G variant may present milder symptoms. Compared with patients carrying null variants, carriers of compound heterozygous missense variants may present with a milder phenotype, manifesting as asymptomatic high CK level.

Advance in tic disorder related genes in children / 国际儿科学杂志

Tic disorders(TD), a complex chronic neuropsychiatric disorder characterized by rapid, involuntary, non rhythmic, single or multiple muscle movements or vocal twitches, usually occurs in children aged from 2 to 15 years.TD has various clinical manifestations, which are usually related to various psychopathology or behavioral comorbidities, including attention deficit hyperactivity disorder, anxiety, depression and obsessive-compulsive disorder.The etiology and pathogenesis of TD are still not completely clear.Immune, genetic, neurobiochemical and environmental factors are generally recognized as factors related to the pathogenesis of TD.In recent years, the research on TD and genetic factors has gradually increased, but so far there is no clear conclusion on the pathogenic genes of TD.This paper only discusses the genes related to TD.

Advance in genetics of lissencephaly / 国际儿科学杂志

Lissencephaly(LIS)is a group of abnormal cerebral cortical dysplasias caused by the defective migration of neurons and it is characterized by thickening of the cerebral cortex, widening of the gyri and disappearance or shallowness of the sulci.Clinically, the patients often have manifestations such as epilepsy, mental retardation, and developmental delay.At present, there is no specific treatment and most patients have poor prognosis.There is currently no specific treatment, and most patients have a poor prognosis.Recently, with the widespreading clinical application of genetic testing, many disease-causing genes related to the lissencephaly have been discovered, so it is important for us to study its pathogenesis and the mode of inheritance.In this study, we reviewed the recent literature on genes associated with lissencephaly.

Effect of psychological intervention combined with flurbiprofen axetil on the recovery period of laparoscopic surgery for sevoflurane anesthesia / 中国生化药物杂志

Objective The purpose of this study was to investigate the effect of psychological intervention and flurbiprofen axetil on the recovery period of laparoscopic surgery for sevoflurane anesthesia.Methods 100 cases of laparoscopic surgery in the First People's Hospital of Jiande City from January 2015 to December 2016 were selected.They were randomly divided into the control group and the experimental group, each with 50 cases.The induction methods of the control group and the experimental group were the same as those during the operation, and propofol and sevoflurane and remifentanil were used to maintain anesthesia.The experimental group patients before intravenous injection of 2 mg/kg flurbiprofen, psychological intervention nursing, pay attention to the psychological status of patients, strengthen the communication with patients, patients with heart to eliminate negative emotions, relieve the psychological pressure of patients.Results The incidence of restlessness before extubation was 32%in the control group,and the rate of restlessness immediately after extubation was 22%.The occurrence rate of restlessness was 56% at 5 minutes after extubation.In the experimental group, the incidence of agitation at different times was significantly lower than that in the control group, with statistical difference(P<0.05).Compared with the control group, the hemodynamics of the experimental group was more stable, and the incidence of nausea and other adverse reactions was lower, with statistical difference(P<0.05).Conclusion Laparoscopic surgery under sevoflurane anesthesia can reduce the incidence of restlessness in recovery period of a certain degree of psychological intervention combined with flurbiprofen,stable hemodynamics,reduce the incidence of adverse reactions.

Effects of RLMF and RLMP on Expression of TRPV5 in IgA Nephropathy Rat Renal Tissue / 医药导报

Objective To investigate the effects of Rosa Laevigata Michx Flavoid( RLMF) and Rosa laevigata Michx Polysaccharose (RLMP) on expression of TRPV5 in IgA Nephropathy (IgAN) rat renal tissue. Methods Forty Sprague-Dawley rats were randomly assigned to four groups.The rat model of IgA nephropathy was induced by intragastric administration of bovine serumalbumin and injections of LPS and CC14.Eight weeks later,the rats with IgAN were treated with RLMF or RLMP (4 weeks), or normal saline.Rats was sacrificed at thirteenth weeks, and RNA was extracted from the kidney.Expression of TRPV5 in tubulointerstitial tissues were analyzed by fluorescent quantitative RT-PCR. Results After RFLP intervention,the expression levels of TRPV5 were markedly increased (P<0.01) than model control group,while decreased (P<0.05) than normal control group but had no significance with model control group after RFLF intervention. Conclusion TRPV5 expression is decreased in IgAN,and RLMP can adjust TRPV5 expression and improve renal function of IgAN.

Role of IGRA in screening of tuberculosis for children with close TB contacts / 实用医学杂志

Objective To study the feasibility of in vitro release of γ-interferon tests (IGRA) in screening of tuberculosis for children with close TB contacts. Methods 185 children with close TB contacts were detected by IGRA at the pediatric clinic in our hospital. Results In IGRA-positive group, the rate of strong positive PPD (PDD≥15 mm) was 50.9%, which was higher than 9.1% in IGRA-negative group (X2 =37.263, P < 0.00). The morbidity rate for children with close TB contacts was 30.2% in IGRA-positive group, and it was significantly higher than 3.0% in IGRA-negative group (X2 = 28.928, P < 0.00). The sensitivity was 80% and the specificity was 77.6% for IGRA screening in children who had close contacts with TB patients. The sensitivity would be 95.0%, as the test was combined with PPD test. Conclusions IGRA screening in children with close TB contacts can increase the detection rate of tuberculosis and reduce imaging screening.

Effect of anhydrous calcium sulfate whisker on the mechanical function of soft denture liners / 中国组织工程研究

BACKGROUND:Due to limitations of the physicochemical properties of soft denture liner material itself, whisker has been added in the soft lining material in recent years, so as to enhance its mechanical properties. OBJECTIVE:To investigate the effect of different additive amount of anhydrous calcium sulfate whisker on the mechanical function of self-curing soft denture liner. METHODS: There were six groups in this experiment. Anhydrous calcium sulfate whisker at the mass fraction of 0 (control), 1%, 2%, 3%, 4%, 5% was respectively added into self-curing soft liner materials, 10 test specimens in each group, a total of 60 test specimens. The shear bond strength, Shore hardness and tensile strength were detected. RESULTS AND CONCLUSION: With the increasing amount of the anhydrous calcium sulfate whisker, the Shore hardness of the soft lining material was increased continuously, and the tensile strength was increased firstly and then reduced. When 3% anhydrous calcium sulfate whisker was added, the bond strength and tensile strength of soft lining material reached the peak. Taken together, the mechanical properties of the soft lining materials became perfect when 3% anhydrous calcium sulfate whisker was added. These results demonstrate that anhydrous calcium sulfate whisker may affect the mechanical properties of self-curing soft liner.

Effect of different root canal cleaning methods on root canal dentin microleakage / 中国组织工程研究

BACKGROUND:Microleakage between restoration, tooth structure and bonding agent can cause the entry of bacteria and liquid in the mouth into the gap, thereby damaging the bonding interface between the restoration and tooth tissues, and leading to bond failure. Microleakage detection can directly show whether the closure of the root canal of post and core system is good or bad. The severity of microleakage directly affects the restorative effects of post and core. OBJECTIVE: To evaluate the effects of different root canal cleaning methods on the microleakage between the fiber post and root canal dentin. METHODS: Thirty fresh non-caries premolar posts with free root canalin vitro were randomly divided into five groups, and the root canal wal was respectively washed with saline, 5.25% sodium hypochlorite solution+17% ethylenediamine tetra-acetic acid (EDTA)+saline, 3% hydrogen peroxide solution+5.25% sodium hypochlorite+ saline, 3% hydrogen peroxide solution+2% chlorhexidine solution+saline, and 2% chlorhexidine solution+17% EDTA+saline in different groups. Super-bond C&B adhesive agent was used for bonding fiber post, and the microleakage of each sample was observed under stereomicroscope. RESULTS AND CONCLUSION: The severity of microleakage in the al groups was ranged as folows: saline group > 3% hydrogen peroxide solution+5.25% sodium hypochlorite+saline group > 5.25% sodium hypochlorite solution+17% EDTA+saline and 3% hydrogen peroxide solution+2% chlorhexidine solution+saline groups > 2% chlorhexidine solution+17% EDTA+saline group.

Effects of different resin cements on the coronal microleakage and bonding strength of fiber posts / 中国组织工程研究

BACKGROUND:At present, there are stil differences in the studies of total-etch, self-etch and self-bonding resin cement effect on the coronal microleakage and bonding strength of fiber posts. OBJECTIVE:To evaluate the coronal microleakage and the bonding strength of fiber posts treated with three kinds of resin cements. METHODS:Total y 32 upper incisors were randomly divided into five groups, including three experimental groups and two control groups. After the root canal preparation, three kinds of resin cements (EMBRACE WetBond, LuxaCore, Medental Multi-cure) were used to fiber posts with the bond diameter of 1.4 mm. Stereomicroscope was used to observe the microleakage. Then, the specimens were cut into 2 mm wafer along the axis of tooth, and universal testing machine for push-out test was used to observe the failure mode. In the positive control group, no root canal preparation was done, the root was coated with nail polish, and the crown was directly exposed to the dye. In the negative control group, no root canal preparation was done, the root canal orifice was covered with the resin, the tooth was overal coated with nail polish and then embedded 1 mm below the section. RESULTS AND CONCLUSION:The microleakage was observed in al the three resin cements, Medental Multi-cure showed the least microleakage and LuxaCore showed the largest microleakage, and there was significantly different among the three kinds of resin cements (P<0.05). The bonding strength of three cements had significant differences (P<0.05), and ranked from high to low:Mdental Multi-cure, LuxaCore, and EMBRACE WetBond. The main fracture modes were binder/fiber post fracture and mixed failure. The results suggest that the total-etch resin cement binds tightly with the dentin, and owns a superiority in the microleakage and bonding property as compared with the self-etch resin cements and self-bonding resin cements.

Neuroprotective effects of 2-BFI on rat model of experimental autoimmune encephalomyelitis and its immunomodulatory mechanism / 中华微生物学和免疫学杂志

Objective To explore the immunomodulatory mechanism of 2-(-2-benzofuranyl )-2-imidazoline(2-BFI) in rat model of experimental autoimmune encephalomyelitis (EAE).Methods Fifty fe-male Sprague-Dawley(SD) rats were randomly divided into five groups by random digit table , including con-trol group(n=10), EAE model group(n=10), low dose 2-BFI group(1.5 mg/kg, n=10), median dose 2-BFI group (3 mg/kg, n=10) and high dose 2-BFI group (6 mg/kg, n=10).The SD rat model of EAE was induced by immunizing with a guinea pigs′spinal cord homogenate ( GPSCH ) .The severity of EAE was scored according to the signs and symptoms .Pathological changes were observed through Hematoxylin-eosin staining, and then the degrees of inflammatory infiltration were evaluated .The number of activated neuroglia that expressed GFAP and iba 1 in lumbar cords was counted by immunohistochemistry .The expressions of IL-1β, IFN-γ, IL-4 and IL-10 in cervical cords were measured by ELISA .Results Compared with EAE group, rats in the low, median and high dose 2-BFI treatment group had lower incidence of EAE , prolonged latency and decreased CNS inflammation , but only the median dose group showed significant alleviation in clinical symptoms and decrease in CNS inflammatory cell infiltration (P<0.05).Immunohistochemical re-sults showed that the numbers of activated microglia were significantly inhibited ,but the numbers of activated astroglia were increased in the rats treated with median dose of 2-BFI in comparison with the EAE group ( P<0.05).Results of ELISA demonstrated that expressions of IL-1βand IFN-γin 3 mg/kg 2-BFI treated group were significantly decreased compared with that in the EAE group (P<0.05), but expressions of IL-4 and IL-10 were remarkably increased(P<0.05).Conclusion 2-BFI at median dose of 3 mg/kg has a benefi-cial neuroprotective effect on rats with EAE , and its mechanism might be related to immunodulation .

Protective effect and mechanism of lipoxinA4 on oxidative injury in trophoblastic cells / 中华围产医学杂志

Objective To explore lipoxinA4 (LXA4) expression in maternal serum of pregnant women and the protective effect and mechanism of LXA4 on trophoblastic cells from oxidative injury. Methods Trophoblastic cells were randomized into six groups: Control group; Lipopolysaccharides (LPS) group, cells were stimulated by 10 μg/ml LPS for 24 h; Intervention group, cells stimulated by LPS were treated with 100 nmol/L LXA4 for 24 h; LXA4 group, cells were treated with 100 nmol/L LXA4 for 24 h; Antagonistic group, cells stimulated by LPS were treated with 100 nmol/L LXA4 plus 100 μmol/L N-tert-butoxycarbonyl-2-pyrrolidine (BOC-2) for 24 h; BOC-2 group, trophoblastic cells stimulated by LPS were treated with 100 μmol/L BOC-2 for 24 h. The serum concentration of LXA4 in normal group and preeclampsia group was detected by ELISA. The intracellular formation of reactive oxygen species (ROS) was detected by 2,7-dichlorofluorescein diacetate (DCFH-DA) as a fluorescent probe. SOD mRNA was analyzed by RT-PCR. SOD and Nrf2 protein expressions were analyzed by Western blot. The levels of SOD in trophoblastic cells were detected by using detection kit. Results (1) The serum concentration of LXA4 was significantly lower in preeclampsia group (165.53±18.89) pg/L than in the control [(545.67±30.91) pg/L, P0.05). Conclusions LXA4 can significantly reduce the oxidative stress of placental trophoblastic cells stimulated by LPS. LXA4 can bind to lipoxin receptors and activate Nrf2-ARE signaling pathway playing a protective effect. So LXA4 in pregnant women can affect the oxidative stress of placenta.

Effect of lipoxin A4 on lipopolysaccharide-induced endothelial hyperpermeability in human umbilical vein endothelial cell / 中华妇产科杂志

Objective To explore whether lipoxin A4 (LXA4)could prevent lipopolysaccharide (LPS)-induced human umbilical vein endothelial cells (HUVEC) monolayer hyperpermeability and its possible mechanism. Methods Human umbilical cords were obtained from women with normal pregnancy immediately after delivery from Tongji Hospital Affiliated of Tongji Medical College. Primary HUVEC were isolated from umbilical veins and subcultured, then, HUVEC were divided into four groups:control group;LPS group (10 mg/L of LPS); LPS + LXA4 group(10 mg/L of LPS and 100 nmol/L of LXA4); LPS +LXA4 + BOC-2 group [10 μmol/L of BOC-2, an effective antagonist of formyl peptide receptor like 1 (FPRL-1)]. All expriments were performed after cells were treated for 24 hours. Endothelial permeability was measured by fluorescein isothiocyan-ate labelled bovine serum albumin (FITC-BSA) clearance across the monolayer; tumor necrosis factor α(TNF-o) mRNA and secretion were detected by reverse transcriplase (RT) -PCR and ELISA assay respectively, and nuclear factor κB(NF-κB) protein change was determined by western blot. Results (1) LPS induced a significant increase in the permeability [Pa value of LPS group was (183.1 ±1.7)%], while co-administrating with LXA4 obviously attenuated this LPS-induced hyperpermeability, Pa value of LPS + LXA4 group was (103.1 ±2.2)%, LPS + LXA4 + BOC-2 group was (162.2 ± 2.8)%, control group was 100%, the permeability of HUVEC monolayer was significantly increased by LPS which was (83.1 ± 1.7)% of control (P <0.01), however, it was notably inhibited by LXA4 (P<0.05); the blockade of FPRL-1 could attenuate the effect of LXA4, that is, there was no difference between the LPS + LXA4 + BOC-2 group and the LPS group. (2) After treatment with different concentration of LPS(0,0.1, 1,10 mg/L), the mRNA expressions of TNF-α were increased (1.11 ±0.11,1.27 ± 0.03, 1.60 ± 0.06, 1.82 ± 0. 04, respectively), compared with the control group, at the concentration of 1,10 mg/L LPS, the difference was statistically significant (P<0. 05). (3) The increased levels of NF-κB and inflammatory mediator TNF-α in the LPS group were both inhibited by LXA4. Levels of NF-κB protein and TNF-o mRNA secretion in LPS treated group (0.53 ±0.06 and 0.81 ±0.09 ,respectively)were both inhibited by LXA4 (0.19 ± 0.05 and 0.41 ± 0.07, respectively, and both had significant difference, P<0.05). (4) Levels of TNF-α in HUVEC culture medium of LPS group [(31.94 ±0.01)ng/L] was significantly higher than the control group [(18.17 ± 0.03) ng/L, P<0.05], LPS + LXA4 group [(15.72 ± 0.07) ng/L] was significantly lower than the LPS group (P<0.05). Conclusion Our findings demonstrated that LXA4 could prevent the endothelial cell hyperpermeability induced by LPS in HUVEC under which the possible mechanism was through inhibiting the expression of NF-κB and its related cytokines through receptor-dependent.

Effect of lipoxin A₄ on IL-1β production of monocytes and its possible mechanism in severe preeclampsia / 华中科技大学学报（医学）（英德文版）

This study examined in vitro effect of lipoxin A(4) (LXA(4)) on interleukin-1β (IL-1β) production of monocytes and its possible mechanism in severe preeclampsia (PE). Peripheral venous blood was drawn from 15 patients with severe preeclampsia (PE group) and 20 normal pregnant women (control group) to prepare monocytes which were then treated with LXA(4) at different concentrations of 0, 10, 100 nmol/L respectively. IL-1β level in the supernatant of monocytes was detected by enzyme linked immunoassay. The [Ca(2+)](i) of monocytes was measured by laser scanning confocal microscopy. The results showed that the IL-1β level and the [Ca(2+)](i) of monocytes in the PE group were significantly higher than those in the control group. LXA(4) significantly decreased the generation of IL-1β in a dose-dependent manner in the PE group. After treatment with 100-nmol/L LXA(4), in the PE group, the [Ca(2+)](i) concentration of monocytes was significantly reduced. It was concluded that LXA(4) may inhibit the IL-1β production of monocytes from severe preeclampsia women by inhibiting extracellular calcium influx.

Effect of lipoxin A4 on interleukin-1β production of human monocytes from severe preeclamptic women and its relationship with cytoplasm free calcium concentration-an in vitro study / 中华围产医学杂志

Objective To investigate the effect of lipoxin A4(LXA4) on interleukin-1β(IL-1β)production of monocytes in maternal peripheral blood from severe preeelampsia women and its relationship with cytosolic free calcium([Ca2+]i)concentration. Methods Peripheral venous blood was drawn from 15 women with severe preeelampsia(SPE group)and 20 normal pregnant women (control group)who were admitted to the First Affiliated Hospital of Wenzhou Medical College from October 2008 to May 2009.Monoeytes were obtained from peripheral blood with Ficolt density gradient centrifugation and then were suspended in RPMI-1640 culture supplemented with LXA4 at the final concentrations of 0,10 and 100 nmol/L,respectively.IL-1β levels in monocytes supernatant were detected by enzyme-linked immunosorbent assay.The cytoplasma[Ca2+]i of cultured monocytes and its variations affected by LXA4 were measured by laser scanning confocal microscope. Results (1) After incubation with different concentrations of LXA4(0,10,100 nmol/L)for 24 h,the levels of IL-1β in SPE group were(63.16±8.20)pg/L,(53.71±8.08)pg/L and(43.16±6.89)pg/L,respectively, indicating a significant inhibition effect on IL-1β level in a dose-dependent manner (P<0. 05). The IL-1β levels in the control group were (19.22±7.43) pg/L, (16.99±6.32) pg/L and (15. 18±5.24) pg/L, correspondingly (P>0.05). (2) Without LXA4, the [Ca2+ ]i concentrations of monocytes in the SPE group were higher than that of the control (1028.09 ± 160. 52 vs 323.61 ±87.86, P<0. 05). After treatment with 100 nmol/L LXA4, the [Ca2+ ]i concentration of monocytes significantly decreased in the SPE group (409.67± 116.73, P<0. 05). Conclusions In vitro LXA4 may inhibit the IL-1β production in monoeytes of SPE patients through decrease of the cytoplama [Ca2+ ]i concentration.

Effect of lipoxin A(4) on IL-1β production of monocytes and its possible mechanism in severe preeclampsia / 华中科技大学学报（医学）（英德文版）

This study examined in vitro effect of lipoxin A(4) (LXA(4)) on interleukin-1β (IL-1β) production of monocytes and its possible mechanism in severe preeclampsia (PE). Peripheral venous blood was drawn from 15 patients with severe preeclampsia (PE group) and 20 normal pregnant women (control group) to prepare monocytes which were then treated with LXA(4) at different concentrations of 0, 10, 100 nmol/L respectively. IL-1β level in the supernatant of monocytes was detected by enzyme linked immunoassay. The [Ca(2+)](i) of monocytes was measured by laser scanning confocal microscopy. The results showed that the IL-1β level and the [Ca(2+)](i) of monocytes in the PE group were significantly higher than those in the control group. LXA(4) significantly decreased the generation of IL-1β in a dose-dependent manner in the PE group. After treatment with 100-nmol/L LXA(4), in the PE group, the [Ca(2+)](i) concentration of monocytes was significantly reduced. It was concluded that LXA(4) may inhibit the IL-1β production of monocytes from severe preeclampsia women by inhibiting extracellular calcium influx.

Effect of lipoxin A4 on lipopolysaccharide-induced oxidant stress in human umbilical vein endothelial cells / 中华妇产科杂志

Objective To explore the effects of lipoxin A4 ( LXA4 ) on lipopolysaccharide ( LPS)-induced oxidative stress in human umbilical veins endothelial cells(HUVEC) and the possible mechanism.Methods Neonatal umbilical cords were obtained from normal term pregnant women with cesarean section within 4 hours and then were used to isolate HUVEC for subculture.HUVEC were divided into four groups:control group; LPS group ( 10 μg/ml of LPS); LPS + LXA4 group ( 10 μg/ml of LPS and 100 nmol/L of LXA4); LXA4 group (100 nmol/L of LXA4) All expriments were performed after cells treated for 12 and 24 hours respectively.Immunofluorescence was used to detect the expression of Ⅷ foctor and nuclear translocation of nuclear factor-erythroid-2-related factor 2 ( Nrf2 ); the mRNA expression of Nrf2, heme oxygenase 1 (HO-1) and reduced form of nicotinamide-adenine dinucleotide quinone oxidoreductase-1(NQO1) were evaluated by reverse transcription-PCR .Results (1)The flavovirens fluorescence was observed in the cytoplasm under fluorescence microscope, which confirmed the existence of Ⅷ factor which specifically expressed in endothelial cells, especially in HUVEC.(2)Immunofluorescent results showed that in control group, Nrf2 protein expressed in the cytosol rather than in the nucleus.In LPS group, the expression of Nrf2 protein obviously increased in the nucleus while decreased in the cytosol after 12 hours.However, after LPS treatment for 24 hours, Nrf2 expression reduced in the cytosol and nucleus.In cotreatment with LPS and LXA4 group,the expression of Nrf2 protein was much higher than that in LPS group after 12 hours or 24 hours.Furthermore, Nrf2 protein also mostly expressed in the cytosol in LXA4 group.(3) After stimulation for 12 hours, compared with control group, the gene expression of Nrf2 and HO-1 were significantly enhanced in LPS group (0.581 ± 0.019 and 0.081 ±0.009, P ＜ 0.05 ) and in LPS + LXA4group(0.692 ±0.048 and 0.136 ± 0.018, P ＜ 0.05 ), the level of NQO1 mRNA in LPS group and LPS +LXA4 group were 0.381 ± 0.009 ( P ＞ 0.05 ) and 0.574 ± 0.034 ( P ＜ 0.05 ).After treatment for 24 hours,compared with control goup, the gene expressions of Nrf2 and NQO1 were down-regulated in LPS group (0.180±0.017 and 0.472 ±0.064, P＜0.05).But in LPS + LXA4 group the expression of Nrf2 and NQOI were upregulated (0.532 ± 0.051 and 0.830 ± 0.068, P ＜ 0.05, compared with treatment for LPS group).The mRNA expressions of Nrf2, HO-1 and NQO1 were increased in LPS + LXA4 group compared with LPS group ( P ＜ 0.05 ).In addition, there was no markedly difference in the expressions of Nrf2, HO1 and NQO1 between control and LXA4 group after 12 hours and 24 hours ( P ＞ 0.05 ) .Conclusion Through activating nuclear translocation of Nrf2 protein from cytoplasm, LXA4 upregulates the Nrf2downstream enzymes, such as NQO1 and HO-1 to protect HUVEC against the oxidative stress induced by LPS.

Clinical characteristics and timing of delivery in women with severe preeclampsia complicated with ascites / 中华围产医学杂志

Objective To investigate the clinical characteristics and the optimal time of delivery in pregnant women with severe preeclampsia complicated with ascites. Methods A retrospective study was conducted on 179 severe preeclampsia mothers and their 195 neonates,presented in the First Affiliated Hospital of Wenzhou Medical College from Jan.2003 to Dec.2005,who were divided into two groups:32 complicated with ascites(ascites group)and 147 without(non-ascites group). The general conditions,mode of delivery and complications including eclampsia,hemolysis,elevated serum level of 1iver enzymes,and low platelets(HELLP syndrome),liver failure,renal failure,heart failure,hypoproteinemia,placental abruption,postpartum hemorrhage and puerperal infection,were also analyzed.Clinical data of all infants(38 from ascites group and 157 from non-ascites group)were analyzed.The incidence and mortality rate of small for gestational age(SGA)in both group within the same gestational age group and those between different gestational age groups in the ascites group were compared. Results (1)The average gestations at admission and delivery in the ascites group were earlier than the other[admission:(32.5±2.1)weeks vs(36.1±3.5)weeks;delivery:(34.1±2.3)weeks vs(37.2±1.5)weeks,P＜0.053.The rate of systemic antenatal care in the ascites group waslowcr than that of the non-ascites group(25.0%vs 53.7%,P＜0.05).More complications werefound in the ascites group than in the non-ascites group(hypoproteinemia:100.0%vs 47.0%;liver and renal failure:31.2%vs 8.2%;HELLP syndrome:9.4%vs 2.0%;postpartum hemorrhage:18.8%vs 2.0%;all P＜0.05).(2)The incidence of SGA in the ascites group was all higher than that in the non-ascites group,however,significant differences was only found between the tWO groups at＞36 weeks(7/9 vs 30.2%,P＜20.05).The perinatal mortalily rates of SGA in the ascites group at＜32 weeks and 32～34 weeks were significantly higher than that in the non-aseites group respectively(＜32 weeks:69.2%vs 19.2%,P＜0.05;32～34 weeks:2/7 vs 0,P＜0.05).(3)The highest perinatal mortality rate and the highest incidence of SGA in the ascites group were found in the groups of＜232 weeks and＞36 weeks,respectively. Conclusions The early onset of ascites and higher rate of complications in severe preeelamptie women implies the adverse maternaI and fetal outcomes.Ascites in severe preeclampsia cases should alert the clinicians.The optimal time for delivery might be at 32～36 weeks of gestations.

Protective effects and its mechanism of lipoxins on human umbilical vein endothelial cells under hypoxia / 中华妇产科杂志

Objective To investigate protective effects and mechanisms of lipoxinA4(LXA4)on human umbilical vein endothelial cells(HUVEC)under hypoxia in vitro.Methods The HUVEC culture were divided into groups as followed:added M199 cudture medium as normal contml groups,added CoCl2 to mimic hypoxia in vitro as hypoxia group and added different concentrations of LXA4(1,10,100 nmol/L)were added to the induced hypoxiai HUVEC as agents intervention group.Morphological changes of HUVEC were observed by using inverted phase contrast mieroscope.The influence of LXA4 on cell survival was investigated by methyl thiazolyl tetrazolium(MTT) assaying method after the treatment with different concentrations of LXA4 and 100 nmol/L lipoxinA4 according to different time(4,8,12 and 24 hours).The expression of von-willebrand factor (vWF) was detected by immunocytoehemistry method. The changes of cytosolic Ca2+ were measured by laser scanning confocal microscope. Results ( 1 ) Morphological changes:the cells under hypoxia lost its normal shapes and showed necrosis, while the cells cocuhured with 100 nmol/L LXA4 were normal appropriately. (2)Survival rate: the survival rates of HUVEC under hypoxia was (40. 1±3.9) % and increased to ( 52. 9 ± 1.4) %, (64. 1 ± 3. 3 ) %, ( 76. 6 ± 1.6) % respectively when added with LXA4 with concentration of 1,10, 100 nmol/L into culture medium. There was significant different survival rate when compared with that of hypoxia group. (3) The level of vWF: The expression of vWF was decreased with the increasing concentrations of LXA4 added into culture medium, the gray values were 203.9 ±0. 7 in 1 nmol/L,204.6 ±0. 9 in 10 nmoL/L,191.8 ±0. 5 in 100 nmol/L respectively, which reached statistical difference in comparison with that of hypoxia groups (P<0. 05). (4) Confocai analysis:the intracellular free Ca2+ concentrations of HUVEC were intensified with LXA4 treatment. Conclusions LXA4 plays an important role in keeping the normal shape of HUVEC under hypoxia, can enhance survival of hypoxial HUVEC and decrease the level of vWF in cytoplasm. The protective mechanism might be via decreasing mitochondria Ca2+ overload and increasing cytoplasm Ca3+ by nucleus Ca2+ transference.

Effect of lipoxins on proliferation and secretion of peritoneal macrophages from patients with precclampsia in vitro / 中华妇产科杂志

Objegtlve To study the effect of lipexins on the proliferation and secretion of peritoneal macrophages from patients with preeclampsia in vitro.Methods Peritoneal macrophages were obtained from 24 patients with preeclampsia(preeclampsia group)and 24 normal pregnant women(normal pregnant group)who were treated in the First Affiliated Hospital of Wenzhou Medical Coilege from March to July 2007.Enzyme linked immunosorbent assay (ELISA) was used to detect the concentration of tumor necrosis factor-α(TNF-α)in the supernatant of macrophages which were pulsed with lipoxins at different concentrations(0,10,100 nmol/L)in both groups after 48 hours.Methyl thiazolyl tetrazolium (MTT)assay was used to detect the inhibition rate of cell proliferation of macrophages which were pulsed with lipoxins at different concentrations(0,10,100 nmol/L)in both groups after 24 hours.Results (1)The concentration of TNF-α:the levels of TNF-α were(1867.5±47.3),(1836.9±4.5) and (1800.5±2.7)ng/L after treatment with differed concentrations of lipoxins(0,10,100 nmol/L)in preeclampsia group vs normal pregnant group[(791.3±62.2),(789.4±2.3),(781.5±1.9)ng/L].The levels of TNF-α in preeclampsia group were significantly higher than that in normal pregnant group(P＜0.05).Lipoxins significantly inhibited the concentration of TNF-α in a dose-dependent manner in preeclampsia group (P＜0.05),while it had no significant effect in normal pregnant group(P＞0.05).(2)Cell proliferation inhibition:Incubation with lipoxins produced a dose-dependent(0,10,100 nmol/L)inhibitory effect on proliferation in preeclampsia group,[(14.8±6.3)%,(32.9±3.6)%,(36.7±3.8)%],vs normal pregnant group[(16.8±6.9)%,(16.7±5.4)%,(15.9±2.1)%].The rate of cell proliferation in preeclampsia group was significantly hisher than that in normal pregnant group.Lipoxins significandy inhibited this growth(P＜0.05),while it had no significant effect in normal pregnant group(P＞0.05).Conclusion Lipoxins can inhibit the proliferation of macrophage and secretion of TNF-α in preeclampsia in a dose-dependent manner.Lipoxins may be potentially useful in prevention and treatment of preeclampsia.

Expression of cytokeratin-19 as a stem cell marker in different-purity islet samples of rats / 中国组织工程研究

BACKGROUND: Pancreatic stem cells can be differentiated into endocrine functioning cells under a suitable cultivation condition, suggesting that pancreatic stem cells can be used as a new source of islet in treating diabetes mellitus.OBJECTIVE: To discuss the expression of stem cell in different purities of pancreatic islets.DESING: A controlled observation based on cells.SETTING: Third People's Hospital of Mianyang City.MATERIALS: The experiment was conducted in the Center Laboratory (Key Laboratory of Chongqing City), the First Affiliated Hospital of Chongqing Medical University from March 2005 to March 2006. Thirty healthy male SD rats, of clean grade, aged 50 days were used in this study. During the experiment, the disposal of animals corresponded to Animal Ethical Standard. Cytokeratin (CK)-19 and β-actin were designed by Shanghai Invitrogen Biological Co., Ltd.Brdu reagent, mouse anti-rat Brdu monoclonal antibody, rabbit anti-mouse (RAM) CK-19 polyclonal antibody and Brdu immunohistochemical kit were from Boster Co., Ltd. (Wuhan); HistostainTM -DS double immunohistochemical staining kit was purchased from Beijing Zhongshan Company.METHODS: Rats were intraperitoneally injected with Brdu for labeling, and subjected to perfusion with Ⅴ-type collagenase via pancreatic ducts, with pancreas being excised, ripped, beaten up, digested and centrifuged to obtain pancreatic islet sediments. After being precipitated, the sediments were divided into 3 groups. Group A: The isolated sediments of islets were not purified; Group B: The sediments were added slowly with 25% Ficoll-400 and Hanks solution for purification; Group C: The sediments were slowly added with 25% Ficoll-400, 11% Ficoll-400 for purification in order.MAIN OUTCOME MEASURES: Expressions of Brdu and CK-19-positive cells in the islet samples were detected by immunohistochemical method, and mRNA expression of CK-19 in the different purities of islets was detected by reverse transcription-polymerase chain reaction.RESULTS: ① The purity of islets in the group A was the lowest. The purity of islets obtained by different purification techniques showed significances among the three groups (F =89.42, P ＜ 0.05). ②Positive expressions of Brdu and CK-19 in the group A were significantly higher than those in the group B and group C (F =18.64, 22.12, 38.61. P ＜0.01). ③Expression of CK-19 mRNA in the group A was significantly higher than that in other groups, with group C showing the lowest.CONCLUSION: Expressions of Brdu and CK-19-positive cells exist in different purities of islets, suggesting the existence of more stem cells in the low purify of islets.