Objective To investigate the role of connexin 43(Cx43)in the apoptosis of pancreatic cancer cell line BxPC and its possible mechanism.Methods pcDNA-Cx43,pcDNA-Cx43N,pcDNA3.0,siRNA-Cx43 and siRNA-NC were transfected into BxPC3 cells via liposome method.Cx43 protein and Cytochrome C(Cyt C)concentration was determined by Western blot,and the apoptosis was analyzed by Annexin V/PI binding assay.The mitechondria apoptosis pathway involved in Cx43 associated apoptosis was examined which contains the depolarization of mitechondrial membrane potential (MMP);fluorospectrophotometer was used to measure the activities of caspase-3 and caspase-9. Gap junction intercellular communication(GJIC) was determined by dye-transfer method.Results Cx43 protein expression increased after BxPC3 transfeetion,apoptosis rate increased from(6.35±0.43)%in empty vector transfection group to(14.29±1.24)%;after H202 treatment,apoptosis rate increased from(20.34±2.47)%to(31.27±2.56)%(P<0.05).Meanwhile,mitochondrial membrane potential was decreased,Cyt C was increasingly released from mitochondria,caspases activities were increased;after siRNA43 interference,apoptosis rate decreased from(7.42±0.47)% to(5.19±1.37)%,after H_2O_2 treatment,apoptosis rate decreased from (19.43±1.71)%to(11.67±1.97)%(P<0.05).Decreased mitochondrial membrane potential and Cyt C release were observed,caspases activities were decreased.GJIC of pcDNA-Cx 43 transfection group increased from 14.52±0.57 to 23.05±3.84.and it increased from 1.70 ±0.24 to 3.84 ±0.45 in the presence of β-GA(P<0.05).But the apoptosis rate was not significantly different.Conclusions Cx43 could promote BxPC3 apoptosis via mitochondrial apoptotic signal pathway,and the possible mechanism included signal pathway other than GJIC.

Objective:To investigate the enantiomer separation for atropine by capillary electrophoresis .Methods:Capillary elec-trophoresis was used with an elastic quartz capillary column (60 cm ×75 mm, effective length of 40 cm).The concentration of phos-phate buffer was 30 mmol· L-1 .The high and time of injection was 10 cm and 5 s, respectively.The detection wavelength was 225 nm.The best separation conditions were selected including the type and concentration of chiral resolving agent , pH of the buffer solu-tion, operating voltage and organic solvent.Results:The optimum conditions of separation were as follows:the pH of buffer solution was 7.0, the concentration of S-β-CDP was 10 mg· ml-1 , and the operating voltage was 12 kV.Conclusion: The method is simple and fast, which can be used to se parate the optical isomers of atrpo ine.

Background Corneal confocal microscopy can denamically offer the morphological image of corneal cells at cellular level.To find out the dynamic change of corneal thickness and morphology of corneal cells with aging is very important for the design of corneal refractive surgery.ObjectivePresent study is to find out the important relevance of dynamic change of corneal thickness and morphology of corneal cells with aging and the design of corneal refractive surgery and investigate the influence of age on central corneal tissue and characterize precisely the anatomy of cornea by in vivo confocal microscopy.Methods122 eyes of 122 normal subjects were enrolled in this study with the age from 18 through 80 years old.The subjects were diagnosed as emmetropia in the Center of Tianjin Medical University from August 2003 to December 2007.All eyes were examined in vivo by confocal microscopy.The cell morphology,cell density and corneal thickness were measured by confocal microscopy.The relationship of central corneal tissue change and age was evaluated.ResultsThe density of corneal superficial basement epithelial cells showed a significantly negative correlation with age (r=-0.355,P=0.017).The keratocyte density in the anterior and posterior stroma indicated a significantly negative correlation with age (r_1=-0.462,P=0.001;r_2=-0.403,P=0.016).The thickness of corneal epithelium cells had a significantly negative correlation with age (P=0.02).The mean value of total corneal thickness and stromal thickness in high myopic eyes was (523.2±26.20)μm and (468.4±20.72)μm respectively,presenting a significantly negative correlation with age (r_1=-0.552,P=0.014;r_2=-0.545,P=0.035).A significantly negative correlation was also found in endothelial cell areas and density with age (r_1=0.417,P=0.004;r_2=-0.598,P=0.002).The percentage of polymorphological corneal endothelial cells was considerably increased(r=0.417,P=0.004)but that of pleomorphism cells was significantly decreased in >60-year-old population compared with younger subjects (r=-0.598,P=0.002).The morphology of corneal basement cell,anterior stroma cytocyte and endothelial cell were normal in younger subjects,and enlargement of the cells in size,decreases of number and density of the cells were found in >60-year-old subject.ConclusionThe anatomy and histology of the central cornea in young subjects exist discrepancy from older ones.The corneal hisological changes with aging include thinner thickness of epithelium,thinner thickness of cornea,decreased density of keratocyte and endothelial cells and increased size of endothelial cells.Corneal thickness is asymmetric and seems to undergo age-related anatomic changes.

Objective:To study the effects of ~(125)I-(α_v)ASODN on the in vitro invasive ability of heptocellular carcino-ma cell line(HepG2) through PEI-RGD-mediated receptor process. Methods: Intergrin α_v-specific antisense oligonucle-otide was labeled with ~(125)I, and PEI-RGD/~(125)I-(α_v)ASODN complex was prepared by combining ~(125)I-(α_v)ASODN with polyethyleneimine derivative PEI-RGD. PEI-RGD/~(125)I-(α_v)ASODN complex was transferred into HepG2 cells through the receptor-mediated process. The effect of PEI-RGD/~(125)I-(α_v)ASODN complex on the invasive ability of HepG2 cells was examined by Boyden chamber invasive assay. Results: (1) The labeling yield and radiochemical purity of ~(125)I-(α_v) ASODN were(73.78±4.09)% and(96.68±1.38)%, respectively, and the labeled compound had a good stability in vitro after 48 h at 37℃; (2) The ability of HepG2 cells to uptake PEI-RGD/~(125)I-(α_v)ASODN reached its peek ([12.77±0.85] % ) when PEI-RGD/~(125)I-(α_v)ASODN was at 4 μl/2 μg ([12.77±0.85] %), and then gredually decreased thereafter. So the dosage of PEI-RGD/~(125)I-(α_v)ASODN for the following experiment was chosen as 2 μl/1 μg; (3) The invasive capacity of HepG2 cells was significantly reduced in PEI-RGD/~(125)I-(α_v)ASODN group compared with those in other experiment and control groups (P <0.01 ). Conclusion: ~(125)I-(α_v)ASODN mediated by PEI-RGD can effectively inhibit the invasive capacity of HepG2 cells.

Effects of different drying methods including sun drying, steamed, boiled, constant temperature drying (at 40, 50, 60 °C) on appearance, hardness, rehydration ratio, dry rate, moisture, total ash, extractive and polysaccharides contents were studied to provide the basis of standard processing method for Tulipa edulis bulbus. The results showed that the treatments of sun drying and 40 °C drying showed higher rehydration ratios, but lower dry rate, higher hardness, worse color, longer time and obvious distortion and shrinkage in comparison with other drying methods. The treatments of 60 °C constant temperature drying resulted in shorter drying time, lower water and higher polysaccharides content. Drying time is shorter and appearance quality is better in the treatment of steaming and boiling compared with other treatments, but the content of extractive and polysaccharides decreased significantly. The treatments of 50 °C constant temperature drying led to similar appearance quality of bulb to commercial bulb, and it resulted in lowest hardness and highest dry rate as well as higher rehydration ratio, extractive and polysaccharides content, moderate moisture and total ash contents among these treatments. Based on the results obtained, 50 °C constant temperature drying is the better way for the processing of T. edulis bulbus.
Color ; Desiccation ; methods ; Plant Stems ; chemistry ; Polysaccharides ; analysis ; Quality Control ; Tulipa ; chemistry ; Water ; analysis

Cutaneous Fistula ; Digestive System Abnormalities ; therapy ; Endoscopy, Gastrointestinal ; methods ; Female ; Humans ; Infant ; Treatment Outcome

Objective To introduce a new economical method which can be used for determination of urinary iodine of batch samples with low cost of arsenic trioxide,and decrease environmental pollution.Methods A catalytic spectrophotometry for measuring iodine content in a small volume of urine samples with low cost of arsenic and cerium was established based on the principle of ammonium persulfate digestion As3+-Ce4+ catalytic spectrophotometry.Ninety-six well polypropylene microplate was utilized as digestion and catalysis reaction vessel,conventional laboratory oven was used as a tool to digest and heat,ice plate was used to cool,and the absorbance was read with the Multimode Reader.The accuracy of the new method was evaluated with the current standard method (WS/T 107-2006) by simultaneous determination of urinary iodine of 24 urine samples.Results The linear range of this method was 0-300 μg/L,the linear correlative coefficient (r) was higher than 0.999,and the detection limit was 8.67 μg/L.The coefficient of variations was 2.15％,4.33％ and 3.48％ when measuring urine samples with high,medium and low iodine concentration,respectively.The test results of three national standard urinary iodine samples were all within the given value range and the relative deviation (RD) was-0.16％,1.81％ and-2.82％,respectively.The average recovery of the low concentration was 97.51％,and that of the high concentration was 96.01％.The two methods correlated well (r =0.995,P ＜ 0.01).Conclusions This method greatly reduces the arsenic waste,environmental pollution,consumables and labor.The new method is simple and efficient,accurate and reliable;it is suitable for application as a supplementary method for analyzing urinary iodine of a large number of samples.

Objective To explore the related diseases,main causes and clinical features of children with basal ganglia calcification(BGC).Methods Thirty cases with BGC detected by CT were studied retorspectively,and its clinical symptoms and image were summarized.Results Many factors and diseases were related to BGC,such as hypothyroidism,intrauterine infection,intrauterine hypoxia,epilepsy,posttraumatic cerebral lacunar infarction.Main clinical manifestations of BGC in children were twich,mental retardation,disorders of limb movements etc.The CT scan showed localized punctuate calcification in basal ganglia.Conclusions The main causes of BGC in children are hypothyroidism,intrauterine infection and intrauterine hypoxia,and the clinical manifestations are diverse.For children with CT-detected BGC should diagnose its causes;and for unknown causes cases should strengthen follow-up.

Objective To investigate the effect of protein inhibitor of activated signal transducer and activator of transcription 1 ( PIAS1 ) gene silencing on the inflammatory response of rat pancreatic acinar cell lines AR42J with cerulein stimulation, to study its role in the pathogenesis of acute pancreatitis.Methods The siRNA targeting PIASI was designed, synthesized, transfected into AR42J cells by lipofectmine 2000.24 h later, cerulean was added and cultured for another 24 h.Subsequent AR42J cells with cerulein stimulation were divided into 4 groups: cerulein, liposome, negative-siRNA and PIAS1-siRNA groups.In addition, a group with PBS was as control group.The activity of p38 mitogen- activated protein kinase (p38MAPK) was detected by western blotting.TNF-α, IL-1β, IL-6, matrix metalloproteinase (MMP) 9 expression were analyzed by RT-PCR and western blotting, respectively.Results The expression of p38MAPK in PIAS1-siRNA, negative-siRNA, liposome, cerulein,and control group was 1.93 ±0.11, 1.22 ±0.10, 1.30 ±0.17,1.32 ± 0.21, 0.12 ± 0.02;while the expression of phosphorylated p38MAPK was 2.10 ± 0.25, 1.36 ± 0.20,1.26 ±0.15, 1.23 ±0.25, 0.58 ±0.48, the expression in PIAS1-siRNA group was significantly increased when compared with other groups (P＜0.05).The levels of TNF-α, IL-1β, IL-6, MMP-9 mRNA were 1.66 ±0.15,1.66 ± 0.15,1.90 ±0.01, 1.56 ±0.20 in PIAS1-siRNA group, while the expression of protein was 2.06 ±0.37,2.20 ±0.34, 1.80 ±0.10, 1.17 ±0.05, which was markedly higher than those in other group (P ＜0.05).Conclusions PIAS1 gene silencing could enhance p38MAPK activity, and improve inflammatory mediator expression in pancreatic acinar cells with cerulein stimulation.

The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water