Objective: To evaluate the specific killing effect of adenovirus (Ad)-mediated double suicide gene system (CD/TK fusion genes) driven by VEGF promoter on pancreatic cancer Capan-2 cells. Methods: VEGF-positive Capan-2 cells were transfected with Ad-VEGFP-CDTK. Ad-free vector acted as negative control. The transfection efficiency was observed and the transcription of CDTK gene was detected by RT-PCR. The Capan-2 cells were treated with 5-fluorocytosine (5-FC) and ganciclovir (GCV) at different concentrations. The effects of the double suicide gene system on cell proliferation and the bystander effects were assessed by MTT assay. Then morphological changes were observed by electronic microscopy and distribution of cell cycle was determined by flow cytometry. Human pancreatic cancer Capan-2 cells were subcutaneously implanted into nude mice. The tumor inhibition rate was calculated. Results: The infection rates of the two resultant recombinant Ads in Capan-2 cells were not significantly different, and they were gradually elevated with the increase in multiple of infection (MOI) of Ads. MTT assay showed that pre-drug dose-dependently inhibited the growth of Capan-2 cells. Apparent bystander effects were also observed. Electronic microscopy demonstrated apoptotic changes of Capan-2 cells. Typical apoptotic peak was detected in double suicide gene system-treated group by flow cytometry. Cell cycle distribution analysis showed that the proportion of G0/G1 increased and the number of cells in G2/M and S phase decreased after treatment. The implanted tumor model was successfully established in nude mice. The tumor size was decreased significantly after treatment with double suicide gene system. Conclusion: The promoter of VEGF regulated double suicide gene system can specifically kill pancreatic cancer Capan-2 cells and induce apoptosis in vitro. And it significantly inhibites the growth of implanted pancreatic tumor in nude mice.

Objective To study the effects of suicide gene system mediated by adenovirus containing the CD-TK fusion gene controlled by human vascular endothelial growth factor(VEGF)promoter on apoptosis of human hepatocellular carcinoma cells HepG2 in vitro.Methods The VEGF-expressing HepG2 cells were infected by adenovirus vector containing CD-TK fusion gene controlled by the VEGF promoter(Ad-VEGFP-CDglyTK).The infection efficiencies in HepG2 cells were observed under a fluorescence microscope.The toxic effect of 5-fluorocytosine(5-FC)and ganciclovic(GCV)on infected cells was determined by light microscopy,electron-microscopy and flow cytometry(FCM).Results The transfection efficiency in HepG2 cells increased with the increasing adenoviral titer.The pro-drug(5-FC and GCV)could induce apoptosis of HepG2 cells in certain range of dosage(the doses of GCV+5-FC:1mg/L + 20mg/L,10mg/L + 40mg/L,100mg/L + 60mg/L)at the multiplicity of infection(MOI)of 100.The effect showed a time-dependent manner.HepG2 cells showed typical morphologic changes of apoptosis after administration of the pro-drug(GCV:10mg/L,5-FC:40mg/L)for 72 hours:chromatin condensation and disposition along nuclear membrane.Karyopyknosis and karyoklasis were seen under electron microscopy(?10 000).Apoptotic peak was also shown in HepG2 cells treated with the pro-drug(5-FC and GCV)by flow cytometry.The cell apoptosis rate was increased accordingly as the concentration of pro-drug(5-FC and GCV)increased.The apoptosis was increased obviously in comparison with the negative control group(P

Objective To evaluate the selectively killing effect of adenovirus(Ad)mediated double suicide gene driven by VEGF promoter on pancreatic cancer cell SW1990. Methods VEGFexpressing SW1990 were infected by Ad-VEGFP-CDTK and Ad-null.respectively.The infection rate was observed and the expression of CDTK was detected by RT-PCR and Western blotting.Followed by treatment with 5-FC and GCV killing effects were evaluated and bystander effects were analyzed by MTF.Pathological character of cells was observed by electron microscopy and distribution of cell cycle was detected by flow cytometry.The caspase-3 activity was detected by absorption spectrometry. Results The infection rate of the resultant recombinant Ad to SW1990 cells was not apparently different.RT-PCR and Western blotting demonstrated product of CDTK gene in SW1990 cell infected by Ad-VEGFP-CDTK.Prodrug could inhibit proliferation of SW1990 and the effect was dose-dependent.There was considerable bystander effect as observed by MTF.Apoptotic peak was also shown by flow cytometry.Morphologic features of apoptosis in SW1990 cells were displayed via electron microscopy.Cells at the G0-G1 phase was increasing and the rate at the G2-M and S phase was decreased by prodrug.The caspase-3 activity gradually rised with the increasing concentration of the prodrug. Conclusions The CDTK fusion gene system controlled by VEGF promoter has killing effect on the VEGF-expressing SW1990 cells and inducing the cell apoptosis.

BACKGROUND: Tissue engineering provides a new way for the repair of urinary tissue and organ defects.Urinary tissue engineering has shown a bright prospect.OBJECTIVE: To review the latest research on urinary tissue engineering at national and international level.METHODS: With the keywords of tissue engineering, urology, scaffold, vascularization in Chinese and in English,respectively, a computer-based search for articles published from January 2000 to January 2016 was performed in CNKI and PubMed databases. The articles addressing urology tissue engineering, scaffolds and vascularization were collected,summarized and analyzed.RESULTS AND CONCLUSION: The selection and cultivation of seed cells, scaffold material performance, tissue construction in vitro, and degree of vascularization all make an important influence on the repair of urinary injuries. As different seed cells hold different biological characteristics, we should make full consideration prior to choosing an appropriate seed cell, so as to pave a good foundation for urinary tissue engineering. Scaffolds with good three-dimensional structure can promote the cell growth and proliferation, tissue in-growth and vascularization.Tissue-engineered materials are superior to traditional repair materials, but still on initial stage, and further large scale trials will be necessary. Moreover, some problems needed to be solved, such as the regenerated tissue with incomplete function different from natural tissues, and regeneration failure caused by biological stent rejection.

A total of 60 isolates of Fusarium were isolated from fruit rot of banana (Musa spp.), papaya (Carica papaya) and guava (Psidium guajava). The most common species recovered from the fruit rot of the three fruit crops were F. semitectum (40 %), F. solani (38.3 %), F. verticillioides (11.7 %) and F. oxysporum (10 %). Fusarium semitectum was isolated from fruit rot of banana, papaya and guava; F. oxysporum from banana and papaya; F. solani from banana and guava and F. verticillioides from banana. From pathogenicity tests, F. solani and F. semitectum were pathogenic to both banana and papaya and F. verticillioides to banana. F. oxysporum was not pathogenic to banana and papaya and F. semitectum was not pathogenic to guava. The results of the present study showed the presence of several Fusarium spp. on fruit rot of banana, papaya and guava and several species are found to be pathogenic causing fruit rot on their hosts.

Drug testing involves many analytical instruments and test items, sample pretreatment is tedious, the industry's intelligence level remains low, making drug testing a labour-intensive job. However, in the era of Industry 4.0 intelligent manufacturing, intelligent transformation of the quality control (QC) laboratory has become the focus of industry. At the same time, driven by consistency evaluation of the quality and efficacy of generic drugs and the centralized procurement policies, pharmaceutical companies have intensified their competition, further stimulating the intrinsic demand for laboratory intelligence. Based on the current state and future trends of the pharmaceutical industry, this review discusses the development of a digital and automated QC laboratory. It points out the necessity of transitioning from the traditional centralized laboratory model to an intelligent, distributed quality control model to accommodate continuous manufacturing processes. At the same time, it also analyses the potential challenges in the implementation process and coping strategies, in order to provide relevant practitioners with ideas for building intelligent QC laboratories.

ObjectiveThe aim of this study was to evaluate and compare the surgical outcomes of endoscop ic and conventional open thyroidectomies in patients with thyroid disease.Methods116 patients with tyroid tumor were enrolled.56 patients underwent endoscopic thyroidectomy ( endoscopic group ),and 60 patients underwent conventional open thyroidectomy( conventional group).We analyzed the patients' clinic characteristics,surgical outcomes and complications between the two groups.ResultsThe blood loss was less in the endoscopic group than the open group[( 16.8 ± 9.6) ml vs ( 24.9 ± 14.2 ) ml,t =- 2.427,P ＜ 0.05].The degree of satisfaction for cosmetic outcome in endoscopy group( 96.4％ ) was higher than that in conventional group ( 16.7％ ) ( x2 =74.508,P ＜ 0.01 ).There was no significant difference in the operating time,volume of drainage and postoperative hospital stay between two groups,and there was no significant difference in the skin ecchymosis,redness and swelling and postoperative pain between two groups( all P ＞ 0.05).No severe postoperative complication was encountered,such as injuries of the re current or superior laryngeal nerve,parathyroid gland injury or massive hemorrhage.ConclusionEndoscopic thyroidectomy has less blood loss,mini-open and excellent cosmetic benefits compared with conventional open thyroidectomy.

Objective To investigate the curative effect of an adenovirus-mediated fusion gene system driven by VEGF promoter (AdVEGF-CDglyTK) on a nude mouse model of colorectal cancer and analyze the mechanism underlying its therapeutic effect.Methods The animal model of the colorectal cancer was established by using transplantation of the cultivated cells,human colorectal cell line LoVo,via subcutaneous injection on the back of nude mice.Twenty nude mice were equally divided into four groups:group Ⅰ received injection of AdVEGF-CDgiyTK plus 5-flurocytosine/ganciclovir(5-FC/GCV);group Ⅱwere given 5-FC/GCV;group Ⅲ were with AdVEGF-CDglyTK;group Ⅳ were used as control.Results CDglyTK was expressed exclusively in the tumor tissues from the group Ⅰ and Ⅲ by RT-PCR.The phenotype and pathological analysis showed that tumor growth was dramatically inhibited in group Ⅰwhen compared with other three groups,while no significant difference was found between group Ⅱ,group Ⅲ and group Ⅳ.The TUNEL assay demonstrated that the apoptosis rate of 38.65% ± 4.20 significantly increased in group Ⅰ when compared with other three groups (F = 397.530,P =0.000).The tumor microvessel density of 3.08±0.79 decreased significantly in group Ⅰ (F = 34.081,P = 0.000) when compared with other three groups.Conclusion The results suggested that AdVEGF-CDglyTK with 5-FC/GCV can inhibit the tumor growth of colorectal cancer significantly in vivo by a mechanism of systeminduced apoptosis and the efficient suppression of angiogenesis.

Objective To investigate the phenotypes and percentages of B 10 cells in different tis-sues from wild-type mice and to identify their biological functions .Methods The percentages of B10 cells derived from different tissues of mice and their responses to lipopolysaccharide ( LPS) stimulation were ana-lyzed by flow cytometry .Magnetic-activated cell sorting ( MACS ) and fluorescence-activated cell sorting (FACS) were used to purify B10 cells, CD4+CD25-T cells and Treg cells.CD4+CD25-T cells and Treg cells labeled by CFSE were co-cultured with or without B10 cells, and then their proliferation were evaluated after 72 h.Results (1) A subset of CD19+CD5+CD1dhigh regulatory B cells was identified in spleen , pe-ripheral blood and lymph nodes from wild-type mice , of which the highest frequency was detected in spleen (3.95%±0.79%, P<0.05).The isolated B cells from different tissues were stimulated by LPS , PMA, ionomycin and monensin (L+PIM) in vitro to express IL-10.Among them, splenic CD19+IL-10+B cells showed the highest expression of IL-10 (P<0.05).(2) Prolonged LPS stimulation (48 h) to CD5+CD1dhigh B cells enhanced the expressions of IL-10 (P<0.01).(3) CD19+CD5+CD1dhigh B cells inhibited the prolif-eration of CD4+CD25-T cells in vitro in a dose-dependent manner (P<0.01), but increased the secretion of IL-10 by CD4+T cells (P<0.01) and the proliferation of Treg cells in vitro (P<0.01).Conclusion Com-pared with other tissues , the percentage of B10 cell subset in spleen is the highest in wild-type mouse , and B10 cells subset can be activated through Toll-like receptor ( TLR ) signaling pathway .The responses of CD4+CD25-T cells and Treg cells in co-culture with B10 cells are regulated by B 10 cell subset through an increased IL-10 production .B10 cells might be a useful cell population for the treatment of inflammatory au-toimmune diseases.

Over 100 viruses are known to cause acute viral encephalitis in human. In order to diagnose a viral central nervous system infection, various laboratory diagnosis methods have been used. In this study, we examined 220 cerebrospinal fluid samples that were received at the Diagnostic Virology Laboratory of University Malaya Medical Centre between year 2004 to 2006, by viral isolation, pathogen specific antibody ELISA, polymerase chain reaction (PCR) and Real-Time PCR. Majority of the samples were from patients <10 years old. Out of 220 samples, 3 were positive for viral isolation, 27 for PCR (inclusive for the 3 positive for viral isolation) and 39 for pathogen specific ELISA. The total positive detection rate of this study was 30%. Herpes virus was the most important aetiologic agent, responsible for 58% of infection, followed by paramyxovirus (especially measles virus) in 26% of infection, and 14% by enterovirus. Parvovirus and flavivirus were the other common viruses. Among the herpes viruses, herpes simplex and cytomegalovirus were the most common.