Objective To investigate the effects of focal mild hypothermia on apoptotic neurons and the expression of glutamate transporter(GLT-1)and metabotropic glutamate receptor(mGluR2/3)after cerebral ischemia in rats.Methods Eighty-eight adult rats were randomly divided into mild hypothermia group,normothermia group and control group.Unilateral permanent middle cerebral artery occlusion(pMCAO)was induced.In mild hypothermia group,the brain temperature of occlusive side was decreased and maintained for 3 h at(33?0.5)℃ with Sticking Focal Mild Hypothermia Instrument.The body temperature of normothermia group was maintained at(37?0.5)℃.3 h,6 h,12 h,24 h,3 d,7 d after cerebral ischemia,the expression of GLT-1 and mGluR2/3 around infarction cortex were determined by Western Blot.6 h,24 h,3 d,7 d after cerebral ischemia,TUNEL method was performed to label neuronal apoptosis.Results 24 h,3 d after cerebral ischemia of mild hypothermia group,24 h,3 d,7 d after cerebral ischemia of normothermia group,the apoptotic neurons around infarction core markedly increased comparing with 6 h after cerebral ischemia(all P

Objective To test whether feeder cells derived from mouse embryonic stem cells(mESCs) could support the growth of mESCs themselves. Methods mESCs were induced to form mouse embryoid bodies(EB),and then fibroblast-like cells were derived from further differentiated mEB(mEB-dF),which served as feeder cells.The undifferentiation of mESCs grown on mEB-dF was confirmed by morphological analysis,colony efficiency and cell differentiation rate of mESCs,immunocytochemistry,alkaline phosphatase staining and RT-PCR.The pluripotency of mESCs grown on mEB-dF was examined by RT-PCR,inducing their differentiation in vivo and in vitro. Results(Forty-eight) fibroblast-like cells lines were derived from the same EB at three periods(d 10,d 15 and d 20),and five of them,mostly derived from d15 EB,were able to maintain mESCs in undifferentiated status and pluripotential ability over 10 passages.mESCs cultured on these feeder cell lines expressed alkaline phosphatase and specific mESCs markers,including SSEA-1,OCT-4,NANOG,and formed EB in vitro and teratomas in vivo.However,the majority of mEB-dF lines(43/48) has no such ability. Conclusion This study not only provides a novel feeder system for mESCs culture,avoiding lot of disadvantages of mouse embryo fibroblasts used as the feeder,but also indicates that fibroblast-like cells derived from mESCs take on different functions.The molecular mechanism of different function of these fibroblast cells is worthy of further investigations.

Pathogen-associated molecular patterns (PAMPs) are conservative molecules associated with groups of pathogens or their products. These molecules are recognized by relevant receptors. PAMPs induce the expression of inflammatory cytokines through the signal cascade. The role of PAMPs in the initiation and development of periodontitis is recently attracting attention. PAMPs induce the expression of inflammatory mediators after they are recognized in the periodontium. This process damages the periodontal soft tissue and osseous tissue, thus resulting in periodontitis. The results of this study will provide an excellent resolution for the treatment of periodontitis by blocking the pathogenic pathway of PAMPs.
Cytokines ; Humans ; Pathogen-Associated Molecular Pattern Molecules ; Periodontitis ; Periodontium

Aim To study the relative bioequivalence of two domestic ciprofloxacin tablets. Methods Therandomized and crossover study was conducted in 18 healthy volunteers.After a single dose of the drugs,their plasma drug concentration was determined using HPLC.Results Both the two domestic ciprofloxacin tablets fit to one compartment model. The main pharmacokinetics parameters of the tested and reference ciprofloxacin were as followings: C_(max):(2.503?0.394) and (2.706?0.579) mg?L~(-1);T_(max):(1.343?0.402) and (1.075?0.379) h;T_(12):(4.174?1.201) and (3.826?1.005) h; AUC_(0-tn): (10.528?2.204) and (10.643?1.922) mg?L~(-1)?h;AUC_(0-∞): (11.409?2.139) and (11.558?2.160) mg?L~(-1)?h;F_(0-tn) and F_(0-∞) was (100.245?18.447)% and (100.470?20.108)%,respectirely. Conclusion The tested and reference formulations are bioeqivalent.

OBJECTIVE:To observe clinical efficacy and safety of Bushen sanhan tongluo decoction combined with moxi-bustion and celecoxib in the treatment of knee osteoarthritis (KOA). METHODS:A total of 70 KOA patients were selected from Chongqing Kanghua Hospital during May 2014-Dec. 2015,and then divided into observation group and control group ac-cording to odd and even number,with 35 cases in each group. Control group was given Celecoxib capsule 0.2 g,qd;observa-tion group was additionally given Bushen sanhan tongluo decoction(one dose a day,300 mL,decocted with water,taking it 3 times in the morning,noon and night)and moxibustion. A treatment course lasted for 4 weeks,and both received 2 courses of treatment. Clinical efficacies as well as TCM syndrome score,VAS score,WOMAC score,lab indexes,joint condition be-fore and after treatment,the occurrence of ADR were compared between 2 groups. RESULTS:Total response rate of observa-tion group (85.71％) was significantly higher than control group (68.57％),with statistical significance (P<0.05). Before treatment,there was no statistical significance in above indexes between 2 groups (P>0.05). After treatment,TCM syn-drome score,VAS score,WOMAC score,erythrocyte sedimentation rate,CRP level and knee swelling score of 2 groups were decreased significantly,compared to before treatment;those indexes of observation group were significantly lower than those of control group,with statistical significance(P<0.05). There was no statistical significance in the number of bone fric-ative joint between 2 groups before and after treatment (P>0.05). There was no statistical significance in the incidence of ADR (5.71％ vs. 2.86％) between 2 groups (P>0.05). CONCLUSIONS:Bushen sanhan tongluo decoction combined with moxibustion and celecoxib can improve clinical symptoms,relieve joint pain,joint inflammation and swelling of KOA pa-tients with good safety.

Objective To investigate the effects of rigid gas permeable contact lens (RGPCL) wearing 3 years on ocular surface in keratoconus.Design Retrospective case series.Participants 73 patients with keratoconus.Methods From July 2001 to July 2004,73 patients (142 eyes) wearing RGPCL for more than 3 years were collected in Peking University Optometry & Ophthalmology Center.Be- fore and at 1 year,2 years and 3 years of RGPCL wearing,density and morphologic changes of corneal endothelium were examined with non-contact specular microscope.Eye axial length,central and peripheral thickness of cornea were measured with A-scan pachymeter. All eyes were examined with slit-lamp microscope periodically.Main Outcome Measures Corneal endothelial density and morphology, corneal thickness,eye axial length,ocular surface changes.Results Before and at 1 year,2 years and 3 years of RGPCL wearing,aver- age corneal endothelial density was 2901.92?445.20,2862.78?497.13,2854.71?526.80,3015.61?421.22 (cells/mm~2)without significant difference statistically (F=1.571,P=0.20).Morphologic changes were not significant during 3 years.Eye axial length was 25.15?1.50, 24.93?1.36,24.78?1.25,25.39?1.31 (mm) without significant difference statistically (F=2.218,P=0.10).Corneal central thickness was 489.09?59.64,484.02?60.80,496.61?59.74,487.44?54.25(?m)without significant difference statistically (F=0.991,P=0.40).Peripheral thickness changes were also not significant statistically during 3 years.69 eyes with mild conjunctival congestion,12 eyes with corneal fluorescence stain and 6 eyes with corneal epithelial rough were found with slit-lamp microscope during follow-up.Conclusions For the patients with keratoconus,long term of RGPCL wear will not lead to significant ocular changes or severe ocular complications.

OBJECTIVETo assess the prevalence rate of Helicobacter pylori (H. pylori) in blood serum, its affecting factors and isoforms (CagA,VacA )infection in the elderly people in Beijing.

METHODS2006 residents were investigated through household questionnaire in different areas of Beijing (urban, suburban and mountainous district), who were older than 60 years old. Serum H. pylori CagA, VacA and Ureas antibody was detected by immunoblotting.

RESULTSThe total H. pylori infection rate was 83.4% and the infectious rate of I form pathogenic H. pylori was 56.0%. The incidence rate in urban or suburban district was higher than that of in mountainous district (P < 0.001). I form H. pylori infection rate in people with heavy labor or young elderly were higher than that of intelegent work or older elderly (P < 0.05 ). I form H. pylori infection rate in people of low diet was higher than that of high protein diet (P < 0.001).

CONCLUSIONThe rate of H. pylori infection in blood serum was high among the elderly people in Beijing with most of it belonged to type I . However, significant differences were noticed on the distribution of isoforms in different age groups, areas, professions and diet habit.

Age Distribution ; Aged ; China ; epidemiology ; Diet ; Female ; Helicobacter Infections ; blood ; epidemiology ; Helicobacter pylori ; classification ; physiology ; Humans ; Male ; Occupations ; Seroepidemiologic Studies

(P < 0.05 ).Scanning electron microscopy showed that the three-dimensional structure of mix-biofilm in control group was more complex.As real-time PCR showed,after 6-hour culture,the expressions of icaA,fbe,aap,hwp1, als3 and efg1 genes in Farnesol group were down-regulated compared to those in control group and after 24-hour culture the expressions of aap,hwp1,als3 and efg1 genes in Farnesol group were down-regulated compared to those in control group.Conclusion With the intervention by fungal quorum sensing molecules Farnesol,the structure in control group became denser and more complex than that in Farnesol group.This change may have a closer correlation with the down-regulated expressions of als3,hwp1 and efg1 genes,which are related to the formation of Candida albicans biofilms.

BACKGROUND:Human amniotic membrane-derived mesenchymal stem cells (AMSCs) are considered to be one kind of adult stem cells that can be easily obtained in large quantities without using an invasive method. Because of their low immunogenicity, anti-inflammatory properties, multipotency of differentiation and without ethical issue, human amniotic membrane-derived mesenchymal stem cells have been proposed as a good candidate to be used in celltherapy and regenerative medicine. However, the biological properties and the differentiation capacity of human amniotic membrane-derived mesenchymal stem cells are stil poorly characterized. OBJECTIVE:To establish a practical method for isolation and purification of human amniotic membrane-derived mesenchymal stem cells, and to study the biological characteristics and dopaminergic neural-like celldifferentiation potential of the human amniotic membrane-derived mesenchymal stem cells. METHODS:Human amniotic membrane-derived mesenchymal stem cells were disassociated and isolated from the amniotic membrane by trypsin and col agenase based enzymic digestion, and purified by percol mediated density gradient centrifugation. Expressions of surface antigens and transcription factors of the human amniotic membrane-derived mesenchymal stem cells were determined by flow cytometry and western blot assays. Based on the osteogenic and adipogenic induction, the multipotent differentiation capability of human amniotic membrane-derived mesenchymal stem cells was determined. Induction of neural celldifferentiation of human amniotic membrane-derived mesenchymal stem cells was conducted in Neurabasal conditioning medium with ATRA supplement. Neural cellassociated bio-markers were determined by immunofluoresence staining and confocal microscope. RESULTS AND CONCLUSION:In this study, we performed a practical method to isolate and purify human amniotic membrane-derived mesenchymal stem cells and amniotic epithelial cells simultaneously, with high cells yield. We demonstrated a group of constitutive expressions of neural antigens and embryonic associated transcription factor proteins (OCT-4, SOX-2 and KLF4) in fresh isolated human amniotic membrane-derived mesenchymal stem cells as wel as in human amniotic membrane-derived mesenchymal stem cells after in vitro passage, which suggested that the human amniotic membrane-derived mesenchymal stem cells not only possessed intrinsic tendency to neural celldifferentiation, but also maintained their stem cellcharacteristics after in vitro passage. We stimulated the human amniotic membrane-derived mesenchymal stem cells in the neurobasal-A and B27 based conditioning medium to induce neural celldifferentiation. The induced human amniotic membrane-derived mesenchymal stem cells displayed an up-regulation of expression in panel of neural and dopaminergic associate molecules (β-tubulin III, neuron-specific nuclear protein, tyrosine hydroxylase, glial fibril ary acidic protein, myelin basic protein and nestin) by flow cytometry and immunofluorescence staining, which demonstrated the multipotent differentiation capability and dopaminergic neuron-like differentiation potential of the human amniotic membrane-derived mesenchymal stem cells.

Objective To study the expression of Ca-A/K channel-related molecules glutamate receptor 2 and 1(GluR2/1) in hippocampus tissues of neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods A total of 60 7-day-old Sprague-Dawley rats were randomly divided into sham operation group and HIBD group. Hippocampal tissues were obtained at 0 h, 1 h, 6 h, 24 h, 48 h and 72 h after HIBD. The expression of GluR2, GluR1 and autophagy marker protein Beclin-1, LC3 were detected by Western blot assay. Results Edema and focal softening and necrosis were observed 6 h after HIBD in the brains of neonatal rats. Compared with Con group, at each time point, the expression levels of GluR2 were lower while the levels of GluR1, Beclin-1 and LC3 were higher significantly in HIBD group (P<0.05). The protein levels of LC3, Beclin-1, GluR1 and GluR2 in hippocampus tissues of HIBD group were significantly different among different time points after the estab-lishment of HIBD model (F=10.65~701.14, P<0.01). The protein level of GluR2 was decreased from 1 h to 24 h after HIBD and reached the lowest level at 24 h. The levels of GluR1, Beclin-1 and LC3 were increased at 6 h, plateaued at 24 h and remained there until 48 h. The levels of these proteins returned back to the initial level at 72 h. Conclusions Ca-A/K channel-related mol-ecules GluR2 and GluR1 play important roles in the autophagic cell death of hippocampus tissues in neonatal rats with hypoxic-ischemic brain damage.