Objective The aim of this study was to establish a mouse model of chronic Parkinson ' s disease in-duced by systemic administration of lipopolysaccharide plus 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP), and to study the changes of behavioral manifestation , numbers of dopaminergic neurons in the substantia nigra pars compacta . Methods Twenty C57BL mice were randomly divided into 2 groups:the saline control group and model group .The mice in the model group received three intraperitoneal (i.p.) injections of LPS (0.25 mg/kg), once daily for three consecutive days.Four hours following the final LPS injection , the mice received one subcutaneous injection of low-dose MPTP (25 mg/kg).The mice of control group were injected with the same volume of saline .Eight weeks later, the motor ability of the mice was evaluated by footprint test and rotarod test .The tyrosine hydroxylase ( TH)-positive cells were observed by immu-nohistochemical analysis .Results Compared with the control group , the scores of behavioral test were significantly lower , numbers of TH immunoreactive cells were significantly less in the Parkinson ' s model group ( P<0.05 ) .Conclusions Behavioral manifestation ,number of dopaminergic neurons in the substantia nigra are significantly changed in the mouse models of Parkinson ' s disease produced by repeated injection of LPS plus MPTP , suggesting that this chronic animal model can be used in the experimental study for pathogenesis and therapy of Parkinson ' s disease .

ObjectiveTo investigate the effect of immunological and inflammatory reactions on type 2 diabetic macrovascular disease.Methods120 patients with type 2 diabetes mellitus and 60 health controls were enrolled in this study.The patients were divided into two sub - groups,type 2 diabetes mellitus group and type 2 diabetic macrovascular disease group.The markers of immunological and inflammatory reactions were detected,and the main associated factors were collected and analyzed by one -way ANOVA and logistic regression.ResultsThe level of FBG,PPG,HbA1c,Hs-CRP and TNF-αt [ (9.86 ± 1.79)mmol/L,( 14.45 ±5.48) mmol/L,( 11.43 ±3.25) ％,(6.79 ±3.71 )mg/L,( 1.99 ±0.65) ng/ml] in the group of type 2 diabetic macrovascular disease group were higher than type 2 diabetes mellitus group [ (7,25±0.64)mmol/L,(10.45 t2.89) mmol/L,(8.56 ±1.58)％,(4.72 ±2.39) ag/L,(1.24 ±0.44) ng/ml,P ＜ 0.05 ].The risk factors in type 2 diabetic macrovascular disease were TNF-α,Hs-CRP and HbA1 c.ConclusionsImmunological and inflammatory reactions play a key role in type 2 diabetic macrovascular disease.

Objective To study the relationship of expression of central cortex glucocorticoid receptor (GR) at protein level with GR expression in the liver at protein level and with changes of serum cortisol and adrenocorticotropic hormone (ACTH) following severe closed traumatic brain injury (TBI) in mice. Methods Severe TBI was established in awake mice by using a BIM-Ⅲ biomechanical machine. At 0.5, 2, 8, 24, 48 and 72 hours after TBI, the total cytosolic GR in the cortex and liver were detected with Western blotting. Levels of serum ACTH and cortisol were measured by ELISA technique and radio-immunological assay (RIA) respectively. Results The expression of GR both in the cortex and liver were obviously down-regulated at protein levels at 2-72 hours after TBI and increased slowly eight hours after injury. The GR in the liver showed no recovery at 72 hours after injury and that in the cortex was decreased continually at 24 hours after injury. Serum ACTH and cortisol levels were increased markedly compared with control group, when there were two different peaks in the observation curve.Conclusion There is glucocorticoid resistance both in the central and peripheral tissues after severe closed TBI in the awake mice, which changes in a time-dependent manner.

Objective To investigate the cytidylyl phosphate guanosine(CpG) islands methylation status of E-cadherin (E-cad) promoter region in human ovarian carcinoma cell lines (ES-2,3 AO, SKOV3 ), and the effect of 5-azacytidine-2 '-deoxycytidines (5-Aza-CdR ) on the cell proliferative ability, invasion and the expression of E-cad protein. Methods Methylation specific PCR(MSP) was used to detect CpG islands methylation status of E-cad promoter region in ES-2,3AO and SKOV3 cell lines. After treated with different concentrations of 5-Aza-CdR, morphological changes of cell lines were observed under microscope. The proliferative ability was evaluated by methyl thiazolyl tetrazolium(MTT) assay. E-cad protein expression was detected by western-blot and cellular invasion was investigated by 24-well matrigel invasion chambers. Results Hypermethylatian status of CpG islands of E-cad promoter region was observed in ES-2 and SKOV3 cell lines, but not in 3AO cell lines. After treated with 5-Aza-CdR (0.1,1.0,10.0 μmol/L), ES-2 and SKOV3 cell lines displayed morphological evidence of differentiation. 5-Aza-CdR was found to decrease proliferation as evidenced by cell growth curve , to increase the level of E-cad protein expression (P ＜ 0.01 ), and effectively inhibit the ability of cell invasion(P ＜0.01 ). Conclusions CpG hypermethylation is an important mechanism of E-cad gene inactivation in ES-2 and SKOV3 cell lines. 5-Aza-CdR be found to inhibit proliferation and invasion, and increase the expression of E-cad probably by the inhibition of hypermethylation.

Effective treatments of acute ischemic stroke include the specialist wards (stroke units) with multidisciplinary treatment team receiving treatment and intravenous thrombolytic therapy.However,only a few patients can receive these treatments.Telestroke can improve the thrombolytic rate of acute ischemic stroke on site in hospitals for lacking of specialists and enable stroke care maintained at a high level in rural areas.This article reviews telestroke in the treatment of acute ischemic stroke and the roles of nurses in telestroke.

Objective To study the relationships of serum neuroglobin and neuron-specific enolase level with periventricular hemorrhage-intraventricular hemorrhage ( PVH-IVH) and periventricular leucumalacia ( PVL) in preterm infants. Methods There were 241 cases of preterm infants whose gestational age was less than 34 weeks and were admitted in NICU of Guangzhou Women and Children′s Medical Center, Guangzhou Huadu District Matermal and Child Health Hospital and Dongguan Taiping Hospital from Jan. 2010 to May. 2013, enrolled in the study. The serum level of neuroglobin and neuron-specific enolase were detected within 12 hours and on the 3 d, 7 d, 14 d after birth. Cranial ultrasound was preformed 2~3 d, 1week, 2weeks, 3weeks, and 4 weeks after birth. They also received Cranial MRI examination before discharge or when the correct gestational age reached 40 weeks. All 241 cases were divided into 3 groups ( no brain damage group, PVH-IVH group and PVL group) according to the result of cranial US and MRI. The differences of the serum levels of neuroglobin and neuron-specific enolase among each groups were compared. Results The results of cranial ultrasound and /or MRI showed: 162 cases had no brain damage ( in no brain damage group) , 50 cases had PVH-IVH ( in PVH-IVH group) , and 20 cases had PVL, 9 cases had PVL and PVH-IVH ( both in PVL group) . Within 12 h and 3 d after birth, the serum levels of neuroglobin in PVL group and PVH-IVH group was significantly higher than those in no brain damage group (P<0. 05), and the serum levels of neuroglobin in PVL group were signigicantly higher than those in PVH-IVH group ( P <0. 05 ) . On 7 d and 14 d after birth, the serum levels of neuroglobin were no significant difference between PVH-IVH group and no brain damage group ( P>0. 05 ) , and there were still significantly higher than those in no brain damage group and PVH-IVH group (all P<0. 05). The serum levels of neuron-specific enolase within 12 h and 3 d after birth in PVH-IVH group and PVL group were significantly higher than those in no brain damage group ( P<0. 05 ) , and there were no significant difference between PVL group and PVH-IVH group (P>0. 05). On 7 d and 14 d after birth, the serum levels of neuron-specific enolase in PVL group were no significant difference compared with PVH-IVH group and no brain damage group (all P>0. 05). Conclusion The increased serum levels of neuroglobin and neuron-specific enolase in preterm infants within 12 h and 3 d after birth would have certain clinical significance for judging whether early brain damage and PVL would happen.

ObjectiveTo evaluate the esthetic and long-term effec ti veness of Cerinate porcelain(CE) and Belleglass HP (BGHP) in restoring discolore d teeth. MethodsA total of 1871 discolored teeth from 647 pat ients were restored using two materials. The observation of short-term(0.5 to l year) and long-term (over 3 years) effectiveness were followed up, and evaluat ions were made. ResultsThe short-term successful rates were 8 9.5 % and 96.75 % for BGHP CE groups, the long-term successful rates were 88.3 9 % and 96.14 %, and the difference was statistically significant (P0.005). Clinical failure, which was mainly related to t eeth category, teeth position, type of restoration and material, showed as resto ration discoloration, fracture and deciduous, etc. Conclusion BGHP shows higher shear bond strength and longer existence, but further study is required for esthetic results. CE restorations show better esthetic results and longer existence. Therefore, it is an ideal choice to restore discolored teeth.

Objective To express a mutated insulin gene in HepG-2 cell line to further research of insulin gene therapy. Methods Native human insulin cDNA was obtained from fetus pancreas with RT-PCR. Furin consensus cleavage sequence was introduced into proinsulin cDNA with site-directed mutagenesis (overlap extension PCR), and the new sequence was named as INS/furin. Subsequently, INS/furin was subcloned into the multiple clone sites of plasmid p(G1RE)3BP-1Luc. The new plasmid p(G1RE)3BP-11?furin was identified with the method of enzyme digestion by Hind Ⅲ and EcoR V. HepG-2 cells were transfected with the plasmid p(G1RE)3BP-11?furin by liposome-mediated method. The transfected HepG-2 cells were incubated for 48h in a glucose-containing medium (25mmol/L), and then the conditioned media were collected and HepG-2 cells were harvested respectively. The expression of INS/furin mRNA in transfected HepG-2 cells was examined by RT-PCR, the regained DNA was sequenced and insulin in conditioned media was investigated by radioimmunoassay. Results Two enzymes, Hind Ⅲ and EcoR V, digested p(G1RE)3BP-11?furin, and 2 fragments with length of 260 bp and 4 700bp, were obtained. The 260bp fragment was identified as insulin/furin, indicating that the target gene had been successfully inserted in specific sites. RT-PCR showed that insulin/furin mRNA was expressed in transfected HepG-2 cell, and the regained DNA was confirmed as insulin/furin by sequencing; while insulin was detected by radioimmunoassay in conditioned media. Conclusion The recombinant mammalian expression plasmid p(G1RE)3BP-11?furin has been successfully constructed, and transfected into HepG-2 cells, which therefore may efficiently secrete bioactive insulin.

Objective To investigate the role of the expression of p27 and cyclin E in the pathogenesis of Bowen's disease and squamous cell carcinoma (SCC). Methods The expression of p27 and cyclin E was assessed by immunohistochemical method in 16 patients with Bowen's disease, 53 patients with SCC and 25 normal controls. Results The expression of p27 was lower in Bowen's disease and SCC compared to that in the normal skin, and the expression level decreased with a decline in the differentiation of tumors, i.e. the lowest expression was observed in poorly differentiated SCC, followed by highly differentiated SCC and Bowen's disease. The expression of cyclin E was higher in Bowen's disease and SCC than that in the normal skin, and the expression level increased with a decline in the differentiation of tumors. The expression of p27 was inversely correlated with that of cyclin E among all the cases of Bowen's disease and SCC. Conclusion The abnormal expression of p27 and cyclin E may contribute to the pathogenesis of Bowen's disease and SCC.

Objective To explore the changes and significance of matrix metalloproteinase 3 (MMP3) during wound healing in rat. Methods Wound fluids and dermal pluripotent stem cells (DPSCs) were isolated with routine method from rats, and they were purified and expanded. Wound fluid was collected on the first day to serve as wound microenvironment, then the responses of DPSCs to wound fluids were investigated, and the expression of MMP3 protein in DPSCs was determined by immunohistochemistry staining. Meanwhile, animal models were repruduced with cutaneous incision and suturing, and the animals were respectively assigned to intractable wound group, in which rats received whole body irradiation, and simple wound group, in which no irradiation was given. The rats were sacrificed on 3rd, 5th, 7th, 10th and 14th day posttrauma (n=5 each), and specimens of wound tissue were harvested, fixed with 10% formalin solution. Twenty four hours later, the samples were dehydrated and embeded, then paraffin section were made. Paraffin sections were stained with hematoxylin and eosin (HE) staining. Light microscopy was used to observe the pathological changes in wounds. Five randomly selected fields were observed under a ?40 objective to evaluate the histological features, especially the amount of tissue repairing cells in wounds. Furthermore, MMP3 contents in wound sites were determined by immunohistochemistry assay and image analysis. Results The expression of MMP3 in DPSCs increased significantly after stimulation by wound fluids. MMP3 contents in rats of simple wound group increased significantly, especially in the dermal tissues, and the peak value appeared 5-7 days after trauma. MMP3 contents in rats of intractable wound group were significantly less than those of simple wound group, and the time when the peak value appeared was also delayed till the 10th day after trauma. Conclusions MMP3 may be an important substrate involved in wound healing. DPSCs may participate in the processes of wound repairing via high expression of MMP3.