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

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 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.

Objective:To establish a method based on the iPLEX analysis of MassARRAY mass spectrometry platform to detect multiplex genetic mutations among Chinese lung cancer patients. Methods:We reviewed the related literature and data of lung cancer treatments. We also determined 99 mutation hot spots in 13 target genes, namely, EGFR, KRAS, ALK, FGFR1, FGFR2, FGFR3, PIK3CA, BRAF, PTEN, MET, ERBB2, AKT1, and STK11, which are closely related to the pathogenesis, drug resistance, and metastasis of lung cancer and are associated with relevant transduction pathways. A total of 297 primers comprising 99 paired forward and reverse amplification primers and 99 matched extension primers were designed by using Assay Design in accordance with the mutation label and format requirements of the MassARRAY platform. The detection method was established by analyzing eight cell lines and six lung cancer specimens;the proposed method was then validated through comparisons with a LungCarta kit. The sensitivity and specificity of the proposed method were evaluated by directly sequencing EGFR and KRAS genes in 100 lung cancer cases. Results:The proposed method could detect multiplex genetic mutations in the lung cancer cell lines, and this finding is consistent with that observed using previously reported methods. The proposed method could also detect such mutations in clinical lung cancer specimens;this result is also consistent with that observed by using the LungCarta kit. However, an FGFR2 mutation was detected only by using the proposed method. The measured sensitivity and specificity were 100%and 96.3%, respectively. Conclusion:The proposed MassARRAY technology-based method could detect multiplex genetic mutations among Chinese lung cancer patients. Indeed, the proposed method can be potentially applied to detect mutations in cancer cells.

Objective To study the clinical outcome of patients with fns-like tyrosine kinase-3internal tandem duplication (FLT3-ITD) positive acute myeloid leukemia (AML) treated with allogeneic hematopoietic stem cell transplantation (allo-HSCT) and to explore the potential prognostic factors to patients' survival including transplant types or disease status.Methods A total of 314 AML patients in our center from October 2006 to October 2012 were retrospectively analyzed,among whom,54 patients were defined with FLT3-ITD positive.Survival rates and treatment-related mortality were further analyzed.Results For all 54 FLT3-ITD positive patients,the 3-year overall survival (3-OS) rate was 56％ and 3-year leukemia-free survival (3-LFS) rate was 47％.The outcome of haplo-identical HSCT was similar as that of sibling donors(3-OS rate:60％ vs 54％ ; 3-LFS rate:54％ vs 45％,respectively).There were 47 patients who received transplantation in first complete remission(CR1).The other 7 patients were of disease relapse or in CR2 before transplantation.Not surprisingly,patients in CR1 had better prognosis than those in nonCR1.Conclusions Allo-HSCT is an effective treatment for AML patients with FLT3-ITD positive mutation.The survival outcome of haplo-identical HSCT was comparable with that of sibling donors.Relapse of AML was the dominant factor related to the mortality of FLT3-ITD positive AML patients after allo-HSCT.

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.

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.

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.