BACKGROUND:Up to date,inbred embryonic stem cells(ESCs) line mainly derived from 129 mouse strain and C57BL/6 strain,occasionally from BALB/c mouse strain,however,few reports concerning ESCs lines derived from Ch inese Kunmingmouse strain.OBJECTIVE:To isolate and culture mouse ESCs of kunming species,additionally,to identify its biological properties.DESIGN,TIME AND SETTING:The experiment with cells as observed subjects was conducted in the institute of Urology,First Affiliated Hospital of Nanchang University from May 2006 to June 2007.MATERIALS:Blastocysts of Kunmin mice with 3.5 days of embryonic age.METHODS:Inner cell mass from 3.5 days embryonic age were isolated from Kunming species mice,cultured on feeder layers of primary mice embryonic fibroblasts,and then the cells were isolated and subsequently cultured.MAIN OUTCOME MEASURES:Colony growth was observed and determined by alkaline phosphatase (AKP) staining;The expression of stage-specific embryonic antigen (SSEA)-1,and cell specific genes of c-Myc,Nanog,Oct3/4 and Sox2 were measured by immunofluorescence and RT-PCR;finally,the ability of differentiation in vitro and in vivo was identified.RESULTS:The ESCs-like colonies presented typical morphological characteristics of ESCs,which was positive to AKP and SSEA-1,and could express several cell specific genes,such as c-Myc,Nanog,Oct3/4 and Sox2,and differentiate into various cell types in vitro and in vivo.CONCLUSION:An ESCs line is successfully dedved from Kunming mice,which has typical biological characteristics of ESCs.

Abstract: In order to investigate immune protection against swine-origin influenza virus (S-OIV) A H1N1, the helper-dependent adenovirus vector (HDAd) system was exploited to construct recombinant HDAd encoding hemagglutinin (HA). The HA gene was synthesized and cloned to the HDAd backbone. Then, the HDAd/HA DNA molecules were transfected into 293Cre4 cells with calcium phosphate. The cells were infected by helper virus 16 hours after the transfection. The 293Cre4 cells were coinfected with HDAd/HA and the helper virus for large-scale preparation of HDAd/HA. The HDAd/HA was obtained and purified twice with CsCI density ultracentrifugation and observed morphologically under transmission electron microscope, and the expression of HA protein was analyzed with RTPCR. Recombinant HDAd/HA expressing HA protein was successfully constructed which could pave the way for in vivo investigation on immunogenicity and efficacy against S-OIV A H1N1 infection.
Adenoviridae ; Cell Line ; Cloning, Molecular ; Genetic Vectors ; Helper Viruses ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; Humans ; Influenza A Virus, H1N1 Subtype

Objective To lay the foundation of new botanical pesticides by screening the insecticidal efficacy of 10 commonly used TCM pesticides.Methods Slide immersion method, trace intravenous drip method and insect body dipping method were respectively used to measure contact action of water extract and ethanol extract of the 10 commonly used TCM pesticides toTeranychus cinnbarinus,Brevicoryne brassicaeandMythimna separate, with herbal extracts to three kinds of insect pests of corrected mortality as observation indexes,Results Alcohol extracts for three kinds of insect pests were found to have contact action, and water extracts had no obvious effect. The Stemonae Radix alcohol extract had obvious contact action on three kinds of insect pests, and corrected mortality rates were 78.7%, 85.9%, and 96.7%, respectively; ethanol extracts of Angled Bittersweet, Sophorae Flavescentis Radix, Xanthii Fructus and Kansui Radix showed good contact action to Teranychus cinnbarinus,. The volume ratio of compatibility among Angled Bittersweet, Sophorae Flavescentis Radix, Xanthii Fructus and Kansui Radix was 2:1:1.5:1.5:1.5:1.5. The insecticidal activity to Teranychus cinnbarinus, was stronger than extracts of single herbs, and corrected mortality was 96.5%. Alcohol extracts of Stellera, hellebore, white mustard seed, and Xanthii Fructus showed good contact action toBrevicoryne brassicae. When the volume ratio of compatibility among Xanthii Fructus, Stemonae Radix, Stellera, and hellebore was 1:1.5:2:2, the activity toBrevicoryne brassicae increased. and corrected mortality was 93.1%.Conclusion The insecticidal activity will be enhanced after TCM extracts are under compatibility.

A method was developed for the determination of 54 volatile hydrocarbons in workplace air by thermal desorption/gas chromatography-hydrogen flame ionization detector. The workplace air was adsorbed by Tenax-TA thermal desorption tubes, then desorbed by thermal desorption and detected by gas chromatography. The experimental results indicated that the coefficients efficiency of 1,1-dichloroethylene, dichloromethane, trans-1,2-dichloroethylene, cis-1,2-dichloroethylene, 2,2-dichloropropane, bromochloromethane, 1,1,1-trichloroethane, 1,2-dichloroethane, 1,1-dichloropropene were 0.9941-0.9986. The detection limits of bromochloromethane, dibromomethane, trichloromethane, bromodichloromethane, 2,2-dichloropropane, dibromochloromethane, bromoform were 5.4-10.3 ng, the minimum detectable concentration was 0.01-0.1 mg/m~3 (the air volume=0.5 L). The coefficients efficiency of other 38 volatile hydrocarbons was above 0.999, the minimum detectable concentration were 0.001-0.01 mg/m~3. The detection limits of alkenes were 0.4-2.7 ng, alkanes 1.4-3.7 ng, aromatic hydrocarbons 0.2-1.0 ng and naphthalene 2.2 ng. The desorption efficiencies of 54 volatile hydrocarbons were 92.1%-113.1% and the relative standard deviations(RSDs) were 0.6%-17.4%. Except for the RSD values of cis-1,2-dichloroethylene, 1,1-dichloroethane, 1,1,1-trichloroethane, 1,1-dichloroethylene, 2,2-dichloropropane, trichloromethane, trans-1,2-dichloroethylene, dichloromethane, bromochloromethane were 5.1%-17.4%, those of other volatile hydrocarbons were below 5%;The experimental results indicated that the breakthrough capacities of 9 volatile hydrocarbons were 400-4000 ng, those of the other volatile hydrocarbons were above 10 μg. Except for the loss rates of 2,2-dichloropropane, bromodichloromethane were 10%-15% in stable experiment, those of other volatile hydrocarbons in Tenax desorption tubes were below 5%, which indicated that 54 volatile hydrocarbons stored in Tenax tubes were stable. The method is a quick and accurate for the detection of volatile hydrocarbons in workplace air.

Objective To compare analgesic and anti-inflammatory effects of Gentianopsis Paludosa and Sophora Alopecuroide of different kinds of compatibility on the mice.Methods Mice were randomly divided into normal control group, positive medicine group, and different kinds of compatibility groups, 10 mice in each group. Mice received gavage for successive 7 days, once a day. Glacial acetic acid twist body method and hot board to pain method were used to observe and compare analgesic effects of different kinds of compatibility. Xylene to ear swelling method and agar to granulation swollen method were used to observe and compare inflammatory effects of different kinds of compatibility.Results Compared with normal group, compound Gentianopsis Paludosa had obvious analgesic and anti-inflammatory effects (P<0.01). When the ratio of Gentianopsis Paludosa and Sophora Alopecuroide was 20:0.6 and 20:0.9, the analgesic and anti-inflammatory effects were the best.Conclusion Gentianopsis Paludosa and Sophora Alopecuroide have good analgesic and anti-inflammatory effects, among which, when the ratio of Gentianopsis Paludosa and Sophora Alopecuroide is 20:0.6, the best effects show.

AIM: Recombinant human adenovirus-p53 injection (rAd-p53) is the first marketed gene therapeutic drug worldwide. This study aimed to evaluate the safety and primary efficacy of rAd-p53 administrated on advanced solid tumors. METHODS: 24 patients with advanced solid tumor treated with rAd-p53 were reviewed, including 5 cases of renal carcinoma, 4 of nasopharyngeal carcinoma, 4 of colorectal carcinoma, 2 of melanoma, 1 of non-small-celllung cancer, 1 of esophageal carcinoma, 1 of gastric cardia carcinoma, 1 of thymic carcinoma, 1 of duodenal carcinoma, 1 of thyroid carcinoma, 1 of pancreatic carcinoma, 1 of endometrial carcinoma and 1 of rhabdomyosarcoma. RAd-p53 was weekly administrated at the dose of 1?10~ 12 VP, and 4 times of administration was defined as one cycle. Administration approach included intratumoral injection,intrabronchial drop in, intraperitoneal injection, intra-arterial infusion and intravenous drip. Combined therapy was given with chemotherapy in 18 cases, radiotherapy in 2, concomitant chemotherapy and radiotherapy in 1, abdominal thermotherapy and orally gefitinib in 1, cytokine immunotherapy in 1 and without combination therapy in 1. RESULTS: 23 cases underwent 35 cycles of therapy except for 1 case discontinued because of early progression. Among the 21 evaluable cases 5 PR, 5 SD and 11 PD were observed. Overall response rate was 23.8%(5/21) and disease control rate was 47.6%(10/21). Grade I-II injection site pain, chill, fever and myalgia were the most frequent side effects. Grade III fever developed in 2 cases and grade III-IV myelosuppression in 4 cases combined with chemotherapy. Furthermore, severe ostealgia occurred in 2 cases and transient hypotension in 1. CONCLUSION: RAd-p53 is tolerable in patients with advanced solid tumor. A further randomized clinical trial is necessary to confirm the antitumor activity of rAd-p53 combined with conventional strategies.

Objective To investigate the mechanism of Gentianopsis paludosa xanthone(GPX)combined with probiotics in the intervention of colon inflammation-tumor transformation in rats by regulating TGF-β1/Smads pathway and inflammatory factors.Methods Ninety rats were divided into the normal group,the model group[drinking sodium dextran sulfate(DSS)for 3 days]and the intervention group by random number table method.The model group was subdivided into the inflammatory stage group,the pre-inflammatory cancer group(DMH injection for 4 weeks),the intermediate inflammatory cancer group(DMH injection for 13 weeks)and the advanced inflammatory cancer group(DMH injection for 21 weeks).The administration group was subdivided into the groups(after the first day of drinking DSS,drugs for each group were given by gavage once a day for 8 weeks)on the basis of the advanced inflammatory cancer group,including the GPX group(GPX 69.3 mg/kg),the probiotic group,the combined group(GPX+probiotics 400 mg/kg)and the thalidomide group(thalidomide 13.5 mg/kg).The disease activity index(DAI),colon length and wet mass index were compared between all groups.Characteristics of colon tumors were observed,and pathological changes of colon were observed by HE staining.The expression levels of transforming growth factor(TGF)-β1,Smad4,Smad7,interleukin(IL)-6 and tumor necrosis factor(TNF)-α were detected by Western blot assay and enzyme-linked immunosorbent assay,respectively.Results Compared with the advanced inflammatory cancer group,the administration groups showed an increase in colon length,the expression levels of TGF-β1 and Smad4 protein,a decrease in colon wall thickness,wet mass index,maximum tumor diameter,the levels of Smad7,IL-6,TNF-α,and DAI score decreased in the GPX group and the combined group(P＜0.05).The structure and morphology of intestinal mucosa were improved in the GPX group,the probiotic group and the combination group,and the structure of colonic crypt and goblet cell number were increased.Compared with the probiotic group and the GPX group,the colon wall thickness,colon wet mass index and tumor number were decreased,the protein expression levels of TGF-β1 and Smad4 were increased,and levels of IL-6 and TNF-α were decreased in the combination group(P＜0.05).Conclusion GPX combined with probiotics could inhibit the transformation of colon inflammation-tumor,and the mechanism may be related to the regulation of TGF-β1/Smads pathway and the inhibition of pro-inflammatory factors of IL-6 and TNF-α.