Objective To evaluate the effects of mechanical stretch on pentraxin-3(PTX-3)mRNA and protein expression in human alveolar epithelial cells (A549 cells).Methods The human lung epithelial adenocarcinoma cells A549(A549 cells)were purchased from cell biology laboratory,Tongji Medical College,Huazhong University of Science and Technology.The cultured A549 cells were inoculated on collagen Ⅰ BioFlex plates and divided into 5 groups(n=3 wells each):group Ⅰ normal control;groupⅡsham mechanical stretch;group Ⅲ mechanical stretch;groupⅣsiRNA and groupⅤ siRNA+mechanical stretch.In group Ⅲ the cells underwent square cyclic mechanical stretch for 4 h using the Flexercell Systcm.In group Ⅳ the cells were transfected with chemosynthetic PTX-3 specific siRNA by RNAi technique.In group Ⅴ at 24 h after being transfected with PTX-3 siRNA the cells underwent mechanical stretch for 4 h.In groupⅡ mechanical stretch of the cells were prevented by Flexstep.The expression of PTX-3 mRNA in the cells was detected by real-time PCR and the expression of PTX-3 protein in the culture media was determined by Western blotting.Apoptosis of the cells was measured bv flow cytometry(Beeten-Dickinson,USA).Results PTX-3 mRNA and protein expression was signlficantly up-regulated by mechanical stretch in groupⅢand decreased by transfection with siRNA in group Ⅳand Ⅴ as compared with group Ⅰ andⅡ(P＜0.05 or 0.01).The apoptosis ratio was significantly higher in group Ⅲ and Ⅴ than in groupⅡ and was significantly lower in group Ⅴ thanin group Ⅲ(P＜0.01).Conclusion Mechanical stretch can up-regulate PTX-3 mRNA expression in A549 cells.

Purpose The research was conducted to evaluate the stability of N-terminal site-specific PEGylated uricase. Methods The enzyme activity is used as an index to evaluate the thermal stability (at 4-80 ℃) , the pH stability, the ability of avoiding the trypsin digestion and half-life in vivo of PEG-uricase, then it is compared with uricase. Results PEGylated uricase is thermally more stable than uricase between 4 ℃ and 60 ℃, but at 70 ℃, the enzyme activity of both PEG-uricase and uricase decreases sharply. In the test of pH stability, the curve of uricase shows an obvious decrease of enzyme activity at 5 .2-6.0 and 9.2-10.0. The behavior of PEG-uricase indicates that the destabilizing was prevented effectively by PEGylation. The test of anti-trypsin digestion suggests that PEG-uricase retains 70% of its enzyme activity,but uricase only 20% ,200 minutes after being reserved in trypsin solution. Stability in vivo indicates that the half-life of PEG-uricase is 1 530 min and uricase, only 45 min. Conclusion PEGylated uricase has improved thermal stability, the pH stability, ability of protecting from trypsin digestion and stability in vivo.

Objective To study the phase correlation of cutaneous multichannel impedance measurements and the contractions, peristalsis of stomach and the conduction properties in space.Methods A spiral gastric body model was rotated to simulate shrinkage and creep of the stomach.In order to verify the feasibility and validity of the multichannel impedance gastric dynamic measurement, experiments with emulation of multichannel impedance gastric motility measuring principle were conducted through three-dimensional simulation on the electrical characteristics of gastric body model in saline tank simulator.Results The emulation results showed that the change of epigastric current density and potential distribution is consistent with the course of moving from top to bottom when lateral edge of model (namely the intumescent location when the stomach is full) after detecting electrodes in sequence.Meanwhile, the measurement voltage wave forms clearly showed the contraction location of stomach after the model rotating for three cycles in a row, and the phase relationship between adjacent measurement channels was also measured.The sixty seconds data acquired by the experiment device proved that voltage wave form of three measurement channels corresponded to the phase relationship of movement status of the gastric body model.Conclusions Cutaneous multichannel wave forms prove a cyclical movement of stomach, and phase difference among the multichannel signals relating to speed of peristaltic wave, position of the measuring electrodes and channel spacing, etc.This study provides simulation and experimental basis for explanation of quantitative study of conduction properties in space and phase relationship between the results of surface multichannel impedance measurements and the contractions, peristalsis of stomach.

Objective To verify the effectiveness of electrical impedance tomography (EIT) with multi-layer electrodes in gastric emptying measurement by simulation experiments using EIT with three-layer electrodes.Methods Firstly,EIT system with three-layer electrodes and gastric emptying measurement simulation equipment was designed and constructed to simulate gastric emptying process and obtain EIT reconstruction images.Secondly,the region where the electrical characteristic of gastric changes was selected as region of interest.Finally,the relative impedance change rate in region of interest for every image obtained from all three layers was consecutively recorded,then the gastric emptying curve was formed and gastric emptying time was calculated.Results There are obvious differences among the sharp of impedance change curves of interested region for three layers,and the gastric emptying time and process of each layer is also different from one another.It is suggested that the passing path and emptying manner of food in gastric emptying process reflected by EIT images varied from layer to layer.Therefore,the necessity and feasibility of using EIT with three-layer electrodes in gastric emptying measurement study is verified.Conclusions EIT with three-layer electrodes can provide more abundant information of gastric motility function associated with gastric emptying pathology and physiology state.It is likely to be a noninvasive,convenient and effective imaging method for the detection and evaluation research on gastric emptying and gastric motility.

Objective Practical electrical impedance tomography (EIT) system with certain measurement resolution (MR) and signal-to-noise ratio (SNR) differs a lot from ideal simulation conditions in EIT methodology research. The aim of this paper was to study the impact of practical system with different MR and SNR on EIT. Methods Based on the ideal simulation boundary voltages of adjacent excitation and adjacent measurement pattern, practical systems whose MR were 0.1 mV and 0.01 mV, SNR ranged from 40-80 dB and no noises were simulated, simulation study for three imaging models A, B and C with different positions in the to-be imaged field under practical system conditions above was carried out using the combined Tikhonov-NOSER regularization algorithm, and error of reconstruction (ER) function and structure similarity (SSIM) function were adopted for quantitative evaluation of image effect. Results There are differences between images obtained under different MR and SNR for different image models. In order to obtain images for three models, SNR of system should be at least 40-50 dB when MR was 0.01 mV. For the goal to obtain high quality images of A, B and C, SNR should be 80, 70 and 60 dB respectively. When MR was 0.1 mV, SNR for obtaining images of A, B, C were 60, 50 and 40 dB. Moreover, the improvement of images with increasing SNR under MR of 0.1 mV was not as obvious as that of MR at 0.01 mV. Conclusions Blind pursuit of single high SNR is of no help to improve system performance and image reconstruction effect except increasing the difficulties in hardware design. The extremity SNR is 50-60 dB for system with MR of 0.01 mV and 60-70 dB for system with MR of 0.1 mV.

OBJECTIVETo explore clinical efficacy of Yiguanjian Decoction (YD) combined Adefovir Dipivoxil Tablet (ADT) in treating HBeAg negative chronic viral hepatitis B (CVHB) active compensated liver cirrhosis (LC) patients.

METHODSTotally 68 HBeAg negative CVHB active compensated LC patients initially treated were assigned to the treatment group and the control group using random digit table, 34 in each group. Patients in the control group took ADT alone, 10 mg each time, once per day. Those in the treatment group additionally took YD, one dose per day. The therapeutic course for all was 48 weeks. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) were detected once in every two weeks. Hepatitis B virus (HBV)-DNA and four items of serum liver fibrosis [procollagen type I (PCN), hyaluronidase (HA), procollagen III peptide (PCIII), laminin (LN)] were detected once per every 4 weeks. Abdominal ultrasound B was performed before and after treatment. The inner diameter of the portal vein and the size of spleen were recorded. The fibrosis degree of liver was evaluated using Fibroscan. Efficacy of Chinese medicine (CM) was evaluated between the two groups before and after treatment using CM syndrome integrals. Efficacy of Western medicine (WM) was also evaluated between the two groups using Child-Pugh grading. Results Compared with before treatment in the same group, ALT and AST levels restored to normal levels, HBV-DNA turned negative (HBV-DNA < or = 1 x 10(2)) in the two groups after 48-week treatment. Besides, levels of TBil, ALB, PCIV, HA, PCIII, and LN obviously decreased (P < 0.05, P < 0.01). Results of ultrasound B showed the inner diameter of the portal vein and the size of spleen decreased. Fibroscan results showed that the elasticity value of the liver obviously decreased (P < 0.05). Besides, post-treatment levels of PCIV, HA, PCEJ, and LN, and the elasticity value of the liver decreased more obviously in the treatment group than in the control group (P < 0.01). There was no statistical difference in post-treatment levels of ALT, AST, TBil, ALB, inner diameter of the portal vein, or the size of spleen between the two groups (P > 0.05). Compared with before treatment in the same group, scores of Chinese medical syndrome and Child-Pugh scores decreased in the two groups after treatment (P < 0.05, P < 0.01). Besides, scores of Chinese medical syndrome decreased more obviously in the treatment group than in the control group (P < 0.05). The effective rate was 8824% (30/34) in the treatment group, higher than that of the control group [67.65% (23/34)] with statistical difference (P <0.05). Conclusion Combined treatment of YD and ADT could significantly improve symptoms of CM and fibrosis degree of liver of HBeAg negative CVHB active compensated LC patients.

Adenine ; analogs & derivatives ; therapeutic use ; Alanine Transaminase ; blood ; Antiviral Agents ; therapeutic use ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; DNA, Viral ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; Humans ; Liver Cirrhosis ; drug therapy ; virology ; Organophosphonates ; therapeutic use ; Tablets

Objective To investigate the inhibitory effect of 5-fluorouracil (5-FU) on hepatocellular carcinoma (HCC) cell growth and to elucidate its potential molecular mechanism.Methods Real-time PCR analysis was conducted to determine the expression of miR-373 in HCC tissue specimens and HCC cell lines.The expression of miR-373 was also evaluated in HepG2 cells after 5-FU treatment.Western blot analysis was performed to detect the protein levels of PPP6C,a verified target of miR-373,with transfection of miR-373 mimics or 5-FU treatment.A rescue assay was conducted to investigate the cell growth in HepG2 cells by using CCK-8.Results miR-373 expression was up-regulated in both HCC tissues and cell lines.miR-373 expression depicted about 2.94-fold augment in HepG2 cells as compared to normal liver cells control (P ＜0.01).5-FU treatment led to a significant decrease of miR-373 levels (approximately 50％,P ＜0.01,48 h) and resulted in a marked increase of PPP6C protein (approximately 2.1-fold,48 h) in HepG2 cells.The overexpression of miR-373 could prevent the impact of 5-FU treatment on cell growth in HepG2 cells and CCK-8 assay showed that HepG2 cell growth was rescued approximately 81％ and 84％ at 24 h (P ＜ 0.05) and 48 h (P ＜ 0.01),respectively.Conclusion 5-FU can repress endogenous miR-373 level,which activates the expression of downstream targeted gene PPP6C,thereby exerting an inhibitory effect on HepG2 cells.

Objective To develop and test a high precision medical device which allows measurement of bioimpedance in regions below skin surface,and to enable the assessment of strained muscle's situation and instruct patient's rehabilitation exercise.Methods The system was based on the four-electrode impedance measurement mode and designed by using impedance converter network analyzer AD5933 and monolithic operational amplifier AD844.The hardware consisted of an excitation current source,a response signal adjustment circuit,a digital demodulation circuit and a shield driving circuit.This hardware circuit was combined with a LCD screen and a keyboard,which turned it into a portable precision bioimpedance testing device.Results Using an model of three components as test target,a comparison between the device and the Agilent 4294A precision impedance analyzer was presented.In the range of 1 kHz to 100 kHz,the device showed the similar relative error of the complex impedance's modulus and 1 degree absolute error of the complex impedance's phase.The strained muscle's bioelectrical impedance changes in a clinical case during the convalescent were presented and discussed with the results proven to be clinical meaningful.Conclusions The device is accurate,simple and convenient,which is proved qualified on helping doctors to assess the injury states of the strained muscle and to instruct patient's rehabilitation exercise.

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.

Objective To study the role of Mycoplasma genitalium(Mg)infection in chronic non- bacterial prostatitis.Methods Culture and two PCR systems were performed on prostatic secretion specimens from 487 patients with chronic non-bacterial prostatitis and 75 health men in Changchun regions. Results positive rate of Mg in the prostatic secretion from the 487 patients was 7.39% (36/487);In the controls,the positive rate of Mg was 1.33%(1/75).The differences between the two groups were statistically significant (X~2=3.88,P