Objective To explore the effects of various dosage and multiple course of Dexamethasone(DEX) on the expressions of WNTs,?-catenin and glycogen synthase kmase-3?(GSK-3?) genes in the lung of premature rats on the 19th day of embryo.Methods Twelve pregnant SD rats were divided into 3 groups randomly:small dose DEX group,large dose DEX group and control group with 4 rats in each group.The rats in control group were injected with saline 9 g/L;rats in small dose DEX group were injected with DEX 0.4 mg/(kg?d),and the rats in large dose DEX group were injected with DEX 0.8 mg/(kg?d) after DEX was diluted to 0.5 mL with saline.On the 19th day of gestation,fetuses were surgically taken out.The reverse transcription polymerase chain reaction PCR method was used to detect expressions of WNT7b,WNT5a,WNT2,GSK-3? and ?-catenin genes mRNA.Results The expressions of WNT7b(0.55?0.19,0.64?0.54)and ?-catenin(2.03?0.58,2.40?0.89)genes mRNA in small dose DEX group and large dose DEX group were significantly higher than those of control group(WNT7b:0.18?0.10,?-catenin:1.77?0.54)(Pa

One hundred and fifty-seven cases of Graves' disease were divided into three groups according to their clinical features and thyroid functions: Group A, untreated; group B, treated but uncontrolled or relapsed; group C, treated and remitted. The positive rates of TSH receptor antibody (TRAb), anti-tnyroglobulin antibody (TgAb), anti-microsomal antibody (TmAb), anti-DNA antibody and anti-nuclear antibody (ANA) were 91.2, 79.4, 79.4, 70.6 and 23.5% respectively in group A. The positive rate of TRAb and TBII were significantly different between group C and group A or B (both P

ObjectiveTo evaluate the effect of low dose of dexamethasone on the blood glucose concentration in patients undergoing craniotomy. Methods20 consecutive patients undergoing craniotomy without a preexisting metabolic disorder were prospectively randomized into 1 of 2 groups: Dexamethasone (group D, n=10) and Normal Saline (group S, n=10), who were given dexamethasone 10 mg intravenous bolus or a saline placebo preoperatively. Arterial glucose concentrations were measured immediately before and after treatment and hourly for 5 hours intraoperatively. ResultsThe arterial glucose concentration in group D increased significantly(F=3.133,P<0.05), while those in group S keep stable. Compared with group S, arterial glucose concentration in group D increased significantly 180 min after dosing(P<0.01). ConclusionIf dexamethasone is used during craniotomy, perioperative blood glucose level should be carefully monitored and controled.

Objective To study the effect of different concentration of 1,25-dihydroxyvitamin D3[1,25(OH)2D3] on cell proliferation,differentiation and the expression of vitamin D receptor (VDR) in mouse MC3T3E1 osteoblast.Methods Osteoblast were cultured in medium with different concentrations of 1,25(OH)2D3.Incubated for 48 h,cell proliferation of osteoblast were examined by MTT reduction assay (mono-nuclear cell direc cytotoxicity assay),the osteocalcin (OC) levels in cell medium were detected by ELISA,and the expression of VDR mRNA and protein were examined by using SYBR Green real-time PCR and Western blot,respectively.Results 1.After incubation with 1,25(OH)2D3 for 48 h,the number of MC3T3E1 osteoblast was significantly less than that in control group(P0.05).3.SYBR Green real-time PCR and Western blot results showed that the expression of VDR mRNA as well as VDR protein of osteoblast in 10-8,10-9 mol/L experimental groups were significantly higher than those in control group (Pa0.05).Conclusions Cell proliferation of mouse osteoblast can be inhibited,while the cell differentiation was promoted by 1,25(OH)2D3.1,25(OH)2D3 up-regulated the expression of VDR in mouse osteoblast,which suggested that the VDR signal pathway may play some role in proliferation and differentiation of osteoblast.

OBJECTIVETo explore the mechanism of the eye-acupuncture for treatment of acute cerebral ischemia-reperfusion injury.

METHODSThirty-two healthy SD rats were randomly divided into a normal group, a sham operation group, a model group and an eye-acupuncture group, 8 rats in each group. The rat model of cerebral ischemia-reperfusion was established with thread occlusion method in the model group and the eye-acupuncture group. The eye-acupuncture group was treated by eye-acupuncture at "liver region", "upper energizer area", "lower energizer area" and "kidney region" for 20 min immediately after reperfusion and at 30 min before sampling. No treatment was done in the normal group and the sham operation group, and no thread occlusion was performed in the sham operation group. The Neurologic impairment was scored and the methods of immunohistochemistry staining, western-blotting and real-time fluorescent quantitation polymerase chain reaction (RQ-PCR) were taken to detect the expression of the aquaporin protein 4 (AQP4) and its mRNA in cerebral cortex after reperfusion for 3 hours.

RESULTSThe neurologic impairment score of 1.50 +/- 0.54 in the eye-acupuncture group was significant lower than 2.63 +/- 0.92 in the model group (P < 0.01). The expression of the AQP4 protein by immunohistochemistry and western-blot respectively were 116.33 +/- 10.24 and 0.53 +/- 0.04 in the normal group, 118.97 +/- 12.72 and 0.55 +/- 0.07 in the sham operation group, and 129.30 +/- 18.36 and 0.67 +/- 0.08 in the eye-acupuncture group, with statistical significance compared to 150.88 +/- 15.82 and 0.94 +/- 0.04 in the model group (all P < 0.01), and there were significant differences between the eye-acupuncture group and the normal group (both P < 0.01). The tendency in the expression of AQP4 protein and its mRNA in all the group were almost the same.

CONCLUSIONThe eye-acupuncture therapy can relieve the cerebral ischemia-reperfusion injury and the protective mechanism is related to the downregulation of the cerebral AQP4 expression.

Acupuncture Therapy ; Animals ; Aquaporin 4 ; genetics ; metabolism ; Brain ; metabolism ; Brain Ischemia ; complications ; genetics ; metabolism ; surgery ; Disease Models, Animal ; Eye ; anatomy & histology ; Female ; Gene Expression ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; genetics ; metabolism ; therapy

OBJECTIVEMultiple signal transduction pathways, for example, Wnt signal transduction pathway, fibroblast growth factor (FGF), bone morphogenetic protein (BMP) etc, are involved in rat fetal lung development. Wnt signal has been shown to play important roles in regulating cell differentiation and proliferation. It is demonstrated that antenatal dexamethasone (DEX) use can induce lung dysplasia. A rat premature delivery model was developed in this study to compare the effects of DEX on antenatal rat fetal lung morphogenesis and the expressions of Wnt7b, beta-catenin and glycogen synthase kinase 3beta (GSK-3beta) genes in the lung of offspring on 19th day of embryo.

METHODSTwelve pregnant rats were divided into three groups randomly: small dose DEX group, large dose DEX group and control group with 4 rats in each group. The rats in the control group were injected with saline 0.5 ml/d; those in small dose DEX group were injected with DEX 0.4 mg/(kg.d), the rats in large dose DEX group were injected with DEX 0.8 mg/(kg.d), DEX was diluted to 0.5 ml by saline. On the 19th day of gestation, the fetuses were surgically taken out and the histologic structures of fetal rat lungs were observed with light microscope. The RT-PCR and Western-blot methods were used to detect the expressions of Wnt7b, GSK-3beta and beta-catenin genes mRNA and protein.

RESULTS(1) Changes of histologic structure included alveolar numbers: small dose DEX group (15.6 +/- 2.1), large dose DEX group (13.2 +/- 1.6), control group (20.8 +/- 2.0); thickness of alveolar septum: small dose DEX group (11 +/- 5) microm, large dose DEX group (11 +/- 4) microm, control group (13 +/- 7) microm; alveolar space: small dose DEX group (2483 +/- 1336) microm2, large dose DEX group (2924 +/- 1705) microm(2), and control group (1913 +/- 764) microm(2). All the parameters showed significant difference between DEX groups and control group. (P < 0.01 for all comparisons). (2) The expressions of Wnt7b (0.55 +/- 0.19, 0.64 +/- 0.54) and beta-catenin (2.03 +/- 0.58, 2.40 +/- 0.89) genes mRNA of the study groups were significantly higher as compared with those of the control group [Wnt7b (0.18 +/- 0.10), beta-catenin (1.77 +/- 0.54)] (P < 0. 05 for all comparisons) while the expressions of GSK-3beta (1.0 +/- 0.5) were lower than those of the control group (1.1 +/- 0.6) (P < 0. 05). The expressions of GSK-3beta protein in cytoplasm of the study groups [(26.6 +/- 19.7) microg, (10.7 +/- 7.4)microg] reduced gradually while beta-catenin's [(79.5 +/- 1.2) microg, (148.3 +/- 30.4) microg] in the nucleus enhanced simultaneously compared with the control group [(50.0 +/- 00.0) microg ].

CONCLUSIONSSmall dose of antenatal DEX usage can improve the fetal lung development, larger dose of DEX may have negative effect on rat fetal lung morphogenesis. Antenatal DEX usage can change the expressions of Wnt7b, GSK-3beta and beta-catenin genes mRNA and protein, these changes may result in paramorphia during pregnancy.

Animals ; Dexamethasone ; pharmacology ; Female ; Lung ; drug effects ; embryology ; metabolism ; Male ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Wnt Proteins ; metabolism

OBJECTIVETo improve the recognition of the clinical features and results of laboratory examination for isolated pulmonary Langerhans cell histiocytosis (PLCH) in children.

METHODThe information of one case with isolated PLCH was analyzed and reports of 11 cases with isolated PLCH were reviewed.

RESULTThe patient we report is only 2 years old with 1 month of course of disease, manifesting with prominent pulmonary involvement: cough and short of breath; CT scan of the chest showed punctiform, nodular and reticular high density opacities involving all lobes of both lungs. Biopsy of the lung tissue showed expression of CD1a, CD68, S-100, consistent with the diagnosis of LCH. He received prednisolone, VP16 and Vindesine with good response. Ten of 11 cases of isolated PLCH reported before manifesting with cough and dyspnea, CT scan of the chest showed interstitial lung changes (5/8), cystic changes (5/8), small nodules (2/8) and pneumothorax (2/8). Langerhans cells were found in 9 cases on lung biopsy, part of biopsy lung tissues were stained with anti-CD1a, the alveolar lavage fluid of the other 2 cases were stained with S-100 and anti-CD1a.

CONCLUSIONIsolated PLCH is rarely reported in children. It manifested with prominent pulmonary involvement: cough and short of breath, and CT scan of the chest showed interstitial lung changes, small nodules or cysts involving the lung, Langerhans cell could be found in lung biopsy, and the immunohistochemical staining in lung biopsy lung and alveolar lavage fluid stained with S-100 and anti-CD1a antibodies.

Biopsy ; Bronchoalveolar Lavage Fluid ; Child, Preschool ; Histiocytosis, Langerhans-Cell ; diagnosis ; pathology ; Humans ; Lung ; pathology ; Male ; Retrospective Studies

This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections ; genetics ; metabolism ; virology ; HEK293 Cells ; Humans ; Nucleoproteins ; genetics ; metabolism ; Phlebovirus ; genetics ; metabolism ; Protein Binding ; Ribonucleoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

OBJECTIVETo study the effect of maternal vitamin D deficiency on lung morphogenesis and platelet-derived growth factor-A (PDGF-A) expression in rat offspring.

METHODSSprague-Dawley (SD) female rats were randomly divided into two groups: normal control and vitamin D deficiency, with 6 rats in each group. The vitamin D deficiecy group was kept away from light and fed with the forage without vitamin D. After 2 weeks, the rats were mated with normal SD male rats. The morphological changes of fetal rat lungs on day 20 of gestation and 1-day-old neonatal rat lungs were observed by light microscope and electronic microscope. The levels of PDGF-A mRNA and protein in fetal and neonatal rat lungs were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) technique and Western blot method respectively.

RESULTSUnder the light microscope, smaller alveolar space, smaller diameter of the respiratory membrane and thicker alveolus mesenchyma were observed in lungs of fetal and neonatal rats from the vitamin D deficiency group compared with the controls (P<0.05). Under the electronic microscope, fewer lamellar bodies but more glycogen deposition in intracytoplasm were observed in the lungs of fetal rats from the vitamin D deficiency group compared with the controls. There was an increased number of empty lamellar bodies in neonatal rats from the vitamin D deficiency group. The levels of PDGF-A mRNA and protein in lungs of fetal and neonatal rats from the vitamin D deficiency group were significantly lower than the controls (P<0.05).

CONCLUSIONSMaternal vitamin D deficiency during pregnancy may inhibit the development of lung morphogenesis and PDGF-A expression in late fetal and neonatal rats. The low expression of PDGF-A may be involved in the inhibitory effect of vitamin D deficiency on the lung development.

Animals ; Calcifediol ; blood ; Female ; Lung ; embryology ; pathology ; ultrastructure ; Platelet-Derived Growth Factor ; analysis ; genetics ; metabolism ; Pregnancy ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Vitamin D Deficiency ; embryology ; metabolism