Cell viability of Pichia pastoris was detected by flow cytometry (FCM) with two reagents fluorescein diacetate (FDA) and propidium iodide (PI). Compared with FDA/PI double-stained dot plots and PI single-stained dot plots,the latter could divide dead and living cells into two separate zones,and get the correct proportion. Then PI single-stained method was used to detect the change of cell viability in Pichia patoris fermentation. At glycerol batch and fed-batch phase,little dead cells were detected. At methanol fed-batch phase,cell viability decreased when cell weight increased,and was only 73.8% at 88 h.

Objective To investigate the effect of aurora kinase inhibitor VX-680 on cell apoptosis and Bcl-2 expressions in human bladder cancer T24 cells. Methods T24 cells were cultured and treated with VX-680 at various concentrations and time points in vitro. VX-680-induced apoptosis of T24 cells was calculated by flow cytometry. The morphological change of treated cells was observed by microscopy;Bcl-2 mRNA and protein expression in T24 cells was detected by RT-PCR and Western blot assay , respectively. Results The apoptosis rate of VX-680-induced T24 cells increased in a dose-and time-dependent manner. The increase of apoptosis rate and decrease of Bcl-2 mRNA and protein expression in VX-680-induced T24 cells were in a dose-dependent manner. Conclusion VX-680 can significantly induce the apoptosis of T24 cells by down-regulating Bcl-2expression in a dose-dependent manner.

Background Traction of epiretinal membrane results in macular morphologic change and visual functional impairment in eye with idiopathic epiretinal membrane (IERM),therefore figuring out their relationship is helpful for evaluation of disease prognosis.Objective This study was to observe the morphological change of macula and microstructure,and analyze the relationship between retina thickness,integrity of cone outer segment tips (COST) line in fovea and visual acuity.Methods This was a retrospective case-observational study.Fifty-six consecutive cases diagnosed as IERM in Zhongshan Ophthalmic Center from March 2011 to December 2011 were enrolled in this study,and all the patients showed unilateral IERM with the normal fellow eyes.Sixteen patients were males and 40 were females,with a mean age of (61.05 ± 6.58) years old.Spectral-domain optical coherence tomography (SD-OCT) examination was performed on the eyes,and Macular Cube (512×128) and HD 5 Line program were selected.Mean retinal thickness of central area (＜1 mm diameter),inner ring area (1-3 mm diameter) and outer ring area (＞3-6 mm diameter) of macula and the status of COST line (intact or fractured) was recorded.Mean thickness of whole macular areas and the difference of foveal microstructure were compared,and the correlation between retinal thickness and visual acuity was analyzed using Pearson linear correlation analysis.Related parameters including age,visual acuity,retinal thickness were also compared between the continuous COST lines group and the fractured COST lines group by independent sample t test.Results The flatted or disappeared fovea was seen in IERM eyes on the SD-OCT image.Retinal thicknesses were (446±89)μm,(418±64)μm and (328±34)ttm in the central area,inner ring area and outer ring area of macula in the IERM eyes,exhibiting significant increasing in comparison with (250±22) μm,(319±17) μm and (279±17) μm in the normal fellow eyes (t=13.370,9.523,7.769,all P =0.000).Significantly negative correlations were found between the visual acuity and the central macular thickness,inner ring thickness or outer ring thickness (r=-0.686,-0.653,-0.417,P＜0.05).In the IERM eyes,COST band was intact in 20 eyes and lack in 36 eyes.Compared with COST band intact group,aging,worse vision and increased retinal thickness were seen in the COST band absent group (t =2.109,P =0.039 ; t =-4.093,P =0.000 ; t =6.669,P=0.000;t=5.376,P=0.000;t=4.247,P=0.000).COST band was clear in all the normal fellow eyes on the SD-OCT image.Conclusions Increase of macular thickness and disruption of COST band reflect the visual function damage in IERM eye.Deficiency of COST band on OCT image seems to be an early indication of photoreceptor damage.Incomplete fovea COST band is often seen in older patients.

In this article we analyzed the current development of moxibustion treating rheumatoid arthritis from the usefulness, advancement, synergistic effect as well as the variance between different kind of moxibustion. We concluded that moxibustion was an effective intervention for treating RA, and the methods used in moxibustion were searched in clinic. But the clinical tralls has a long way to go, we should pay more attention to the critical issues while in the use of moxibustion.

Objective To investigate the clinical value of the probe melting curve analysis‐based PCR assay for the detection of three types of αT .Methods A total of 149 blood and prenatal archival DNA samples (6 of which were amniotic fluid samples)with three knownαT genes ,which included 63 carriers with Hb CS ,22 cases with Hb QS ,43 individuals with Hb WS and 1 double heter‐ozygote with Hb CS and Hb WS) as well as 20 samples with normalα‐globin gene sequence that had been confirmed by RBD com‐bined with DNA sequencing were selected to test the specificity of probe melting curve analysis by blind analysis .Results The probe melting curve analysis accurately detected 100 of the DNA samples previously characterized by S RBD combined with DNA sequencing .Conclusion Probe melting curve analysis‐based PCR assay for the detection ofαT is featured with rapidity and accuracy and can be applied to clinical and prenatal diagnosis .

0.05).However,neither CpG-ODN nor hTNF-? failed in improving the expression rates of CD1a.Conclusions CpG-ODN,like hTNF-?,has remarkable im- munostimulatory effect on the differentiation and maturation of monocyte-derived DC from patients with chronic hepatitis B.

Objective To study the clinical diagnosis and treatment of malignant gastrointestinal stromal tumor. Methods To retrospectively analyze the clinical data of 28 cases with malignant gastrointestinal stromal tumors underweat surgical treatment . Results The malignant gastrointestinal stromal tumor in adults were more than 50 years old,71.4%(20/28) ,and common clinical symptoms were gastrointestinal bleeding,anemia,and pain. The lesion site: 19 cases of gastric bowel, 8 cases of small intestine, 1 case of colon, radical excision in 22 cases, local excision palliative resection in 5 cases, three cases were multi-visceral resection. Conclusion Malignant gastrointestinal stromal tumor could be diagnosed by the means of endoscopic imaging and clear,and preoperative diagnosis was difficult. Surgical resection was the pathology diagnosis and treatment of primary method,if necessary,to ensure multi-visceral resection of the tumor to prevent recurrence of thoroughness, had important significance.

The method was established for determination of geniposidic acid, chlorogenic acid, pinoresinoldiglucoside, which are three kinds of constituents of Eucommia ulmoides absorbed into the blood components. LC-MS/MS technique was applied to determine the blood components of the bloodstream after administration of E. ulmoides extract. At the same time, HPLC was used for detection of the ingredients content of the blood sample from 23 batches of E. ulmoides. The results showed that geniposidic acid, chlorogenic acid and pinoresinoldiglucoside are prototype into the blood in rats after oral administration of E. ulmoides extract, The linear range of geniposidic acid, chlorogenic acid and pinoresinoldiglucoside was good, and the average recoveries geniposidic acid, chlorogenic acid and pinoresinoldiglucoside were 98.69%, 100.8% and 98.39%, respectively. The method is simple and feasible with good reproducibility.
Animals ; Chlorogenic Acid ; blood ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Eucommiaceae ; chemistry ; Glycosides ; blood ; Iridoid Glucosides ; blood ; Lignans ; blood ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

Objective To evaluate whether estradiol can inhibit and cure the inflammation of experimental preeclampsia in rats. Methods Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 ?g/kg). Rats with normal pregnancy were infused with sodium chloride solution.A group of preeclampsia rats was injected with 17?-estradiol (17?-E_2, 1 mg?kg -1 ?d -1 ). Blood pressure, albuminuria,inflammation associated adhesion molecule CD_ 49d and tumor necrosis factor-?(TNF-?) were assessed. Results On pregnant day 19, for normal pregnancy group(group C) the blood pressure was (120.4?2.0)mm Hg (1 mm Hg=0.133 kPa),urinary protein (0.47?0.06)mg/24 hours;for experimental preeclampsia group(group A) blood pressure was (134.2?2.4) mm Hg,urinary protein(0.79?0.10)mg/24 hours; for experimental preeclampsia with 17?-E_2 treatment group (group B) blood pressure was(123.3?1.7)mm Hg,urinary protein (0.51?0.08)mg/24 hours. A significant increase of blood pressure and urinary albumin was observed in group A. CD_ 49d expression and TNF-? concentration were also increased. 17?-E_2 reduced the expression of CD_ 49d , concentration of TNF-?,blood pressure and albuminuria of experimental preeclampsia. However, the weight of fetuses in 17?-E_2 treatment group were less than that in other groups. Conclusion 17?-E_2 can improve the symptoms of experimental preeclampsia,but its effects on fetus need to be further studied.

Objective To characterize the antibiotic resistance,homology and carbapenemase genotypes of imipenem resistant Acinetobac1ter baumannii isolated from our hospital,and analyze the clonal relatedness of the test strains.Methods Ninety five strains of imipenem resistant A.baumannii were isolated from August 2003 to December 2004 in the First Affiliated Hospital, College of Medicine,Zhejiang University.The MICs of 16 antimicrobial agents against these strains were determined by agar dilution and E-test method.The homology of these isolates was analyzed by pulse-field gel electrophoresis(PFGE).The coding gene of carbapenemases was amplified.PCR products were purified,cloned and sequenced.Plasmid DNA was extracted and purified.Conjugation and Southern blot were performed to locate the position of oxa 23 gene.Results The resistance rates to ampicillin-sulbactam and cefoperazone sulhactam were 67.9% and 30.2%.Polymyxin E had the lowest resistance rate of 17%. The resistance rate to other antimicrobial agents was higher than 90%.The 95 strains,isolated from 10 clinical units,were classified into 6 clones.Clones A and B were predominant clones.All strains produced carbapenemases which were confirmed as OXA 23 by PCR and sequencing analysis.No plasmid was extracted and conjugation was not successful.Southern bolt showed that oxa-23 gene was located on Apal-digested chromosomal segments about 220 kb and 200 kb in Clones A and B,re spectively.Conclusions OXA 23-producing A.baumannii has become one of the most important multi-resistant pathogens in our hospital.Clones A and B have widely spread in our hospital.Oxa-23 gene is located on chromosomal DNA.