Objective To investigate the effect of isovolumic hemodilution with 6% hydroxyethyl starch (HES) (200/0.5) on expression of intercellular adhesion molecule-1 (ICAM-1) after global cerebral ischemia-reperfusion in rats. Methods Eighty-four male Wistar rats weighing 230-280g were randomly divided into 3 groups: group Ⅱ sham operation (S , n = 20); group ischemia-reperfusion (Ⅱ , n = 32) and groupⅢ hemodilution with 6% HES (200/0.05) (H, n =32) . Group Ⅱ and group Ⅲ were further divided into 4 equal subgroups with 8 animals in each subgroup: 2h, 4h, 8h and 12h after beginning of reperfusion. Global cerebral ischemia was produced by permanent occlusion of bilateral vertebral arteries and cross-clamping of bilateral common carotid arteries for 10min and then clamping was released for reperfusion. In group Ⅲ acute normal volumic hemodilution was performed at 10 min after reperfusion was begun. 1ml/100g of blood was removed from artery and equal volume of 6% HES(200/0.5) was infused into the vein simultaneously. Hct was checked before and after hemodilution. The animals were decapitated at designed time and brain tissue was removed from ischemic area and frozen in liquid nitrogen. ICAM-1 expression was determined by using immunohistochemical technique. Results ICAM-1 expression significantly increased after 2h, 4h, 8h and 12h reperfusion in group Ⅱ and Ⅲ as compared with in group Ⅰ (P

Objective To investigate the effect of hemodilution with different plasma substitutes on the expression of tumor necrosis factor-? (TNF-?) and interleukin-1 (IL-1 ) in brain tissue after global cerebral ischemia-reperfusion (I/R) in rats.Methods 116 male Wistar rats weighing 230-280 g were randomly divided into 4 groups: group S ( n = 20) sham operation; group Ⅰ ( n = 32) I/R; group H (n = 32) I/R + hemodilution with 6% hydroxyethyl starch (HAES, 200/0.5) and group G ( n = 32) I/R + hemodilution with gelatine solution. Group Ⅰ, H and G were further divided into 4 equal subgroups with 8 animals in each subgroup according to the duration of reperfusion: 1, 3, 6, 12 h. Global cerebral ischemia was produced by permanent occlusion of bilateral vertebral arteries and cross-clamping of bilateral common carotid arteries for 10 min. The clamping was then released for reperfusion. Global cerebral ischemia was confirmed by coma, loss of righting reflex, bilateral pupil dilation and loss of pain sensation. In group H and G acute hemodilution was performed at 10 min after the beginning of reperfusion. 1 ml? 100 g-1 of blood was removed from the right femoral artery and equal volume of plasma substitute was infused into left femoral vein simultaneously within 5 min. Hematocrit was checked before and after hemodilution. The animals were decapitated after being reperfused for different periods of time as planned and the brains were immediately removed. MTT bioassay and radioimmunoassay techniques were used to determine the IL-1 activity and TNF-? content of the brain tissue respectively. Results The IL-1 and TNF-? levels of brain tissue at 1, 3, 6, 12 h after reperfusion was started were significantly higher in group Ⅰ, H and G than in group S (P

Objective To construct a database for the genetic polymorphism of 19 STR loci in Han population from Hainan province. To investigate the application of 19 STR loci in the paternity testing. Methods The genotypes of 462 unrelated individuals in Hainan were detected with GoldeneyeTM 20A PCR Amplification Kit. 19-STR database was acquired, analyzed and evaluated in 283 paternity testing cases. Results No deviations of allele frequency from Hardy-Weinberg equilibrium expectations were found for Chi-square test (P>0.05). Observed heterozygosity (Hobs) varied between 0.603 and 0.914, total discrimination power (TDP) of 19 STR loci was more than 0.999999999999999, cumulative probability of exclusion (CPE) for triplet cases was 0.999999994. In all 283 paternity testing cases, triplets and duos were 170 and 113 respectively; there were 36 (12.7%) excluded cases comparing to 247 confirmed cases (87.3%). 14 mutation events were observed, and all were one-step mutation. Conclusion 14 out of 19 loci showed highly polymorphic in Han population from Hainan, and 19 STR system has high cumulative probability of exclusion and can meet the needs of paternity test of the local region. But mutation should be paid special attention to.

OBJECTIVETo observe the effect of Hydroxy Safflower Yellow A (HSYA) on the expression of osteogenic markers, such as alkaline phosphatase, Cbf(alpha)l and type I collagen, and explore the mechanism of HSYA in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head.

METHODSFifteen healthy and adult New Zealand white rabbits were collected and weighted 0.9 to 1.3 kg. The rabbits were injected abdominally with anesthetic drugs, then received marrow cavity puncture of tibia and anterior superior iliac spine to get bone marrow blood. Rabbits bone marrow mesenchymal stem cells (BMSCs) were separated from the bone marrow blood, cultured in vitro and passaged. The 3rd generation of BMSCs which had good growth condition were randomly divided into blank group, model group and HSYA groups with different doses. The BMSCs in model group were treated with high dose of dexamethasone to induce adipogenic differentiation of cells cultured in vitro, and inhibit osteogenic differentiation. The BMSCs in HSYA groups received high dose of dexamethasone and different concentrations of HSYA simultaneously. The blank group received not any special handling. After a week,the expressions of alkaline phosphatase, Cbf(alpha)l and type I collagen mRNA were detected.

RESULTSThe alkaline phosphatase activity was significantly decreased in BMSCs of the model group as compared with the blank group (P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also decreased significantly (P<0.01). The alkaline phosphatase activity was significantly increased in BMSCs of each HSYA group as compared with the model group (P < 0.05 or P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also increased significantly (P < 0.05 or P < 0.01).

CONCLUSIONThe mechanism of HSYA may be related to the effect of antagonism to the reduced osteogenic differentiation induced by glucocorticoid.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chalcone ; analogs & derivatives ; chemistry ; pharmacology ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor alpha Subunits ; genetics ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rabbits

Acupuncture Therapy ; instrumentation ; Adult ; Female ; Humans ; Lactation ; Mastitis ; physiopathology ; therapy ; Young Adult

Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.

BackgroundPterygium is an ocular surface disease of abnormal cell proliferative kind and angiogenesis plays an important role in its development and recurrence.Several anti-angiogenic therapies have been used to treat pterygium,but there very few studtes for the in vivo observation of the microvessles in pterygium.ObjectiveThis study was to observe angiogenesis in pterygium with a high-resolution confocal microscope in vivo and to perform immunohistochemical study in vitro.MethodsA prospective case-controlled study was designed.Twenty eyes of 20 consecutive patients with primary pterygia and 20 age- and sex-matched patients with inner eye diseases and strabismus with normal conjunctiva were enrolled in this study.An in vivo confocal microscopy imaging system (Heidelberg Retina Tomograph Ⅱ Rostock Cornea Module) was used to collect microvascular pictures from the anterior part of pterygia and normal nasal conjunctiva of controls,and then immunochemistry was performed to examine the expression of CD31 in microvessel in vitro.The vascular density values were compared between these two groups.The correlation of vascular density values between in vivo Heidelberg Retina Tomograph and in vitro immunohistochemistry was calculated.Written informed consent was obtained from pationts before any examination and surgery.ResultsUnder the in vivo confocal microscope,the microvessel density was (8929±2993) μm/mm2 and (4202 ±692)μm/mm2,respectively,in pterygium and the normal conjunctiva group with a statistically significant difference between them (t =6.881,P＜0.01 ).Immunochemistry revealed that the expression of CD31 to measure vascular density was ( 21.00 ± 4.06/400 × field ) and ( 6.07 ± 1.75/400 × field ) in pterygium and the normal conjunctiva group,showing significant difference (t =12.312,P＜0.01 ).Positive correlations were found in the vascular density values between in vivo corneal laser confocal microscopy examination and in vitro immunochemistry examination in both the pterygium group and normal conjunctiva group (pterygium group:r=0.649,P＜0.01 ;normal conjunctiva group: r=0.572,P＜0.01 ) ConclusionsIn vivo confocal microscopy imaging is superior to in vitro immunochemistry in evaluating the microvessel of pterygium.The results of this study offer a new way index for further investigation of the biological behavior of pterygium and its mechanism.

Objective To discuss the expression and significance of neutrophil to lymphocyte ratio(NLR) and platelet to lymphocyte ratio(PLR) in peripheral blood in patients with chronic obstructive pulmonary disease(COPD).Methods One hundred and fifty cases COPD patients who were treated in Respiratory Departments of Hai'an People′s Hospital Affiliated to Nantong University from January to December 2015 were selected and divided into stable period group with 60 cases and acute exacerbation period group with 90 cases,and 50 healthy subjects were selected as control group.Clinical data were collected and white blood cell count(WBC),neutrophil count(NC),lymphocyte count(LC),platelet(PLT),NLR,PLR,forced expiratory volume in one second(FEV1%) predicted and C reactive protein(CRP) were tested.Results NLR,PLR and CRP values were significantly higher in patients in acute exacerbation period group than in stable period group and control group(NLR:(10.30±9.93) vs.(6.76±5.90) vs.(5.85±3.67);PLR:(335.88±164.94) vs.(286.80±118.98) vs.(194.11±88.14);CRP: (42.18±24.89) mg/L vs.(23.27±13.59) mg/L vs.(22.49±13.47) mg/L,F=6.637,6.808,11.007,P<0.05),but FEV1% predicted was no difference in stable period group and acute exacerbation period group((58.85±16.83)% vs.(57.47±17.76)%;t=0.228,P>0.05).There was a significant positive correlation between NLR,PLR and CRP in acute exacerbation period group(r=0.374,0.379,P<0.01),there was a positive correlation between NLR and PLR(r=0.482,P<0.01),there was a negative correlation between NLR,PLR and FEV1(r=-0.297,-0.299,P<0.05).There was a significant positive correlation between NLR,PLR and CRP in table period group(r=0.593,0.433,P<0.01),NLR and PLR were also positively correlated(r=0.618,P<0.01),NLR,PLR and FEV1 accounted for the expected value of the negative correlation(r=-0.324,-0.342,P<0.05).Conclusion Both NLR and PLR are inflammatory markers,furthermore they show a significant negative correlation to airflow limitation in patients with COPD.

BACKGROUND: Microglia play an important role in immune surveillance in their quiescent state, but the role of the activated microglia is under discussion. OBJECTIVE: To analyze the mechanism of activated microglia in acute cerebral infarction. METHODS: Totally 96 male Kunming mice were selected and randomly divided into four groups, including transplantation, placebo, blank control and sham operation groups. Permanent occlusion of the middle cerebral artery was performed using suture method in the mice of the transplantation, placebo and blank control groups, followed by injection of microglia suspension via subclavian vein, medium containing the same volume of microglia, and nothing, respectively, at 12 hours after modeling. In the meanwhile, the same amount of microglia suspension was injected into the mice of the sham operation group. The Zea-longa scale and brain-derived neurotrophic factor expression at 12, 24 and 72 hours after modeling, the volume of cerebral infarction and the number of nerve cells positive for microtubule-associated protein-2 at 72 hours after modeling were detected. RESULTS AND CONCLUSION: The Zea-longa scale score was 0 point in the sham operation group, which was significantly lower than that in the other three groups at each time point after modeling (P < 0.01). The Zea-longa scores in the transplantation group were significantly lower than those in the placebo and blank control groups at 24 and 72 hours after transplantation (P < 0.01). The positive expression rate of brain-derived neurotrophic factor in the transplantation group was significantly higher than that in the other three groups after transplantation (P < 0.01). The sham group showed no infarction, while the size of cerebral infarction in the transplantation group was significantly lower than that in the placebo and blank control groups (P < 0.01), and the microtubule-associated protein-2 positive rate was significantly higher than that in the placebo and blank control groups (P < 0.01). These results manifest that the activated microglia can improve the survival rate of nerve cells, promote the recovery of cerebral nerve function and reduce the size of cerebral infarction.

0.05].Sao and Eao was significantly different between CHD group and the control group,but Aao has not significant different.③Sao positively correlated with ascending aortic distensibility coefficient(D)(r=0.73,P=0.03),and negatively correlated with aortic stiffness(?)(r=-0.68,P=0.03).Conclusion:Elastic properties of the aorta can directly be assessed by measuring the movements in the upper wall of the aorta with DTI.Reduced aortic S-velocity is significantly correlated with Ascending aortic distensibility coefficient(D) and stiffness index beta(?),which are important factors in assessing the changes of the aortic distensibility.