This research is for the purpose of comparative analysis of the microbial flora structure in the chilled beef with no packing and cling film, which under the same terms of sale. It was used the V3 area fragment of 16S rDNA to carry on PCR-DGGE, Meanwhile used the 16S rDNA sequence to analysis the microbial flora structure of the two samples, according to the technology of clone .The research discovered that the flora structure displays a biggish difference; there was 6 OTU in the chilled beef with cling film, mainly was that Lactococcus(28%), Lactobacillus (26%), Carnobacterium(18%) and Brochothrix (10%); but there was 18 OTU in the chilled beef with no packing, mainly was that Lactococcus(28%), Brchothrix(18%), Acinetobacter (11%). The result indicates that cling film played a certain inhibitory action regarding the Staphylococcus as well as the cold pole bacteria and such bacterium. And it can provide a certain theory ba-sis for the meat processing in the department of microorganism’s control.

It is used the method of pure culture,Selected 32 strains,which were obvious difference in the shape,color and so on common characteristic,From the chilled beef with no packing and cling film on sale in this research;and it was included 12 strains from the chilled beef sample packed with cling film;20 strains from the chilled beef sample with no packing.Simultaneously selected 4 strains which were predominant in each bacterium from the two samples to conduct the further research,8 strains serial numbers are:S01～S08,S01～S04 from the chilled beef sample with no packing;S05～S08 from the chilled beef sample packed with cling film.Through ARDRA(Amplified ribosomal DNA restriction analysis) as well as 16S rDNA to clarify the bacterium's classified status.The physiological and chemical tests were done to determine the various bacteria respective genus.The experiment indicated:S01 is Pseudomonas putida;S02 is Shewanella cincia stain;S03 and S05 are the same Shewanella putrefaciens;S04 is Stenotrophomonas mal-tophilia;S06 is Psychrobacter;S07 is Staphylococcus sciuri;S08 is Microbacterium-laevaniformans.It was proved that two samples altogether have the same predominant bacterium.It can provide certain theory basis for the chilled meat processing craft as the preliminary investigation in the cultured microorganism situation in two samples.

Objective To observe change of T regulatory cells (Tregs) and its relation with in-jury severity and sepsis following severe muhiple injury. Methods A total of 60 patients were em-ployed in the study and divided into severe group (30 patients) and critical group (30 patients) based on ISS scores and into sepsis group (22 patients) and non-sepsis group (38 patients) based on complication of sepsis. The proportion of Tregs in peripheral blood in different groups was detected by flow cytometry at days 1,3,5 and 8 after injury. Results The proportion of Tregs was significantly increased at day 5 postinjury, with statistical difference compared with that at day 3 postinjury (P < 0.01). The proportion of Tregs remained increasing at day 8 pestinjury (P < 0.05). At day 8 postinjury, the proportion of Tregs in critical group was significantly higher than that in severe group (P < 0.01). At the same time, the proportion of Tregs in sepsis group was significantly higher than that in non-sepsis group (P < 0.05). Spearman correlation analysis showed a positive correlation of Tregs proportion with ISS score (rs =0.654, P < 0.01). Conclusions Tregs play an important role in suppression of T cell-mediated im-munity after severe injury. The variation of Tregs can help evaluate prognosis and predict the risk of com-plicating sepsis in patients with severe multiple injury.

Objective To investigate the roles of phosphatidylinositol 3 kinase (PI3K) and phosphorylated Akt (P-Akt) in the pathogenesis of cervical squamous cell carcinoma and condyloma acuminatum.Methods Immunohistochemistry and Western blot were used to detect the expressions of PI3K and P-Akt in tissue specimens from the lesions of 30 cases of cervical squamous cell carcinoma,30 cases of condyloma acuminatum and the prepuce of 15 normal human controls.The average optical density and gray scale values were calculated and analyzed by t test and F test respectively.Results The expressions of PI3K and P-Akt were observed in only the basal layer of the epidermis of control specimens,but in the whole epidermis of condyloma acuminatum tissue specimens.Cervical squamous cell carcinoma tissue specimens displayed a stronger expression of PI3K and P-Akt compared with the control and condyloma acuminatum tissue specimens.As immunohistochemistry revealed,the average absorbance value for PI3K and P-Akt was 0.28 ±0.05 and 0.20 ± 0.07 respectively in cervical squamous cell carcinoma tissue specimens,0.22 ± 0.04 and 0.17 ± 0.03 respectively in condyloma acuminatum tissue specimens,and 0.16 ± 0.04 and 0.10 ± 0.02 respectively in the control tissue specimens; significant differences were observed in the expressions of PI3K and P-Akt among the three groups of tissue specimens (F =44.87,20.64,respectively,both P ＜ 0.01 ).The results of Western blot were consistent with those of immunohistochemistry,and there was a significant difference in the gray scale value for PI3K and P-Akt between cervical squamous cell carcinoma,condyloma acuminatum and control tissue specimens (3.48 ± 0.48 vs.1.99 ± 0.11 vs.1.00 ± 0.03,F=354.83,P＜ 0.01; 3.33 ± 0.26 vs.1.96 ± 0.11 vs.1.00 ± 0.03,F=302.33,P＜ 0.01 ).Conclusions The PI3K/Akt signaling pathway is abnormally activated in condyloma acuminatum and cervical squamous cell carcinoma,and human papilloma virus may cause the abnormal proliferation of infected epithelium likely by affecting the upregnlated expression of PI3K/P-Akt.

Objective To explore the recuperative effect of immunological function and nutritional status on the patients treated by immune enteral nutrition in early stage after severe multiple injury (SMI). Method The patients with SMI,in department of Trauma Surgery,Tongji Hospital,Tongji Medical College, Huazhong University of Science and Technology,between January 2006 to May 2007 were randomly divided into 2 groups: immune enteral nutrition group (IEN group, 20 cases), enteral nutrition group (EN group, 20 cases). The health persons served as the control group(15 cases) .Since 1st postinjury day, all patients were treated with nutritional support. The T-cell subgroup in periphera blood were detected by FCM and the level of PA, RBP, IL-2 and IL-4 in blood serum were detected by ELASA on the 1st, 3rd,5th, 8th postinjury day. Results After the treatment of IEN and EN,the serum levels of PA, RBP and the proportion of T-cell subgroup were significantly increased on the 8th postinjury day compared with on 1st postinjury day (P ＜ 0.01), but there were no differences between IEN group and EN group. The level of IL-4 were significantly decreased and the level of IL-2 were significantly increased in each group on 8th postinjury day, at same time, the level of IL-2 were significantly increased in IEN group compared with EN group (P ＜ 0.05), and the level of IL-4 were significantly decreased in IEN group compared with EN group (P ＜ 0.05). The duration of SIRS was transient and the infected complication was low on the patients treatment by IEN than EN. Conclusions On the patients with severe multiple injury, IEN was most ascendant than EN to improve the immunosuppression and clinical prognosis.

Objective To study the alteration of microbial population distribution and resistance of clinical bacterial isolates in patients with severity muhiple injuries. Methods The distributed Features of 432 strains of infection germs detected among the patients with severe muhiple iniuries admitted into hospital from January 2004 to December 2006 were statistically analyzed during. Results In the total 432 strains,the G accounted for 62.9%(272/432),dominated mainly by pathogens including Pseudomonas aeruginosa,Acinetobacter baumanni I and Escherichia Coli.The G+accounted for 37%(160/432),mainly including Staphylococcus anreus,enterococci and coagulase negative staphylococcus (CNS).Mixed infection rate was 41.1%.The isolating rate of enterococci.CNS and Sten Matophilia was obviously upgraded. Conclusions The source of infection in patients with severity multiple injuries is Gram-negative bacterium,suggesting that surveillance of bacterial resistance and rational use of antimicrobial agents should be emphasized during clinical therapy.

OBJECTIVETo observe the effects of adipose-derived mesenchymal stem cells (ADSCs) with continous over-expression of glial cell line-derived neurotrophic factor (GDNF) on the motor function recovery and nerve regeneration of sciatic nerve of rats after electrical injury.

METHODSFive SD rats were collected to prepare ADSCs with over-expression of GDNF. One hundred and fifty SD rats were divided into normal control group (N), GDNF-ADSCs group (GA), ADSCs group (A), GDNF group (G), and physiological saline group (P) according to the random number table, with 30 rats in each group. Rats in group N were routinely fed without treatment, and rats in the other 4 groups were inflicted with electrical injury on sciatic nerve of thigh of the right hind leg. Rats in groups GA, A, G, and P were respectively injected with 100 µL suspension of ADSCs with over-expression of GDNF (1 x 10(7) cells per mL), 100 [µL ADSCs suspension (1 x 10(7) cells per mL), 100 µL GDNF solution (100 mg/L) , and 100 µL physiological saline to the surface of the injured nerves immediately after injury. Six rats of each group were collected for measuring hind limb stride from post injury week (PIW) 1 to 8, and morphology of the sciatic nerves was observed in PIW 8. In PIW 4, the protein expression of GDNF of sciatic nerves of the rest rats in each group was determined with Western blotting. Data were processed with one-way analysis of variance, analysis of variance of repeated measurement, and SNK test.

RESULTSCompared with that of group N, the hind limb stride values in groups GA, A, G, and P were significantly lower at each time point (with P values below 0.05). Compared with those of group P, the hind limb stride values in group GA from PIW 3 to 8, in group A in PIW 3, 5, and 7, and in group G in PIW 3, 5, 7, and 8 were significantly longer (with P values below 0.05). The hind limb stride values in group GA from PIW 4 to 8 were respectively (10.83 ± 0.97), (13.25 ± 1.40), (12.86 ± 1.42), (14.06 ± 1.50), and (15.09 ± 1.17) cm, which were significantly longer than those in group A [(8.90 ± 0.82), (9.03 ± 0.57), (9.27 ± 0.36), (9.86 ± 0.36), and (9.52 ± 0.58) cm] and group G [(8.87 ± 0.69), (8.51 ± 1.18), (9.34 ± 0.87), (9.76 ± 0.67), and (9.50 ± 1.22) cm], with P values below 0.05. Compared with that of group N, the number of myelinated nerve fibers of sciatic nerves was obviously decreased in group P but obviously increased in groups GA, A, and G; the diameter of axons was obviously shorter, and the myelin thickness was obviously increased in groups GA, A, G, and P in PIW 8 (with P values below 0.05). The number of myelinated nerve fibers in group GA was 31.2 ± 0.8, which was significantly higher than that in group A (23.7 ± 2.7), group G (22.3 ± 2.7), or group P (9.3 ± 2.8), with P values below 0.05. The diameter values of axons among groups P, A, G, and GA were similar (with P values above 0.05). The myelin thickness of rats in group GA was (3.41 ± 0.34) µm, which was significantly thicker than that in group A [(2.64 ± 0.37) µm] or group G [(2.41 ± 0.34) µm], with P values below 0.05. In PIW 4, the protein expression of GDNF of sciatic nerves was significantly higher in groups P, A, G, and GA than in group N (with P values below 0.05), and the protein expression of GDNF in group GA was significantly higher than that in group P, A, or G (with P values below 0.05).

CONCLUSIONSADSCs over-expressing GDNF protein can obviously promote the motor function recovery and nerve regeneration of sciatic nerve of rats after electrical injury.

Adipose Tissue ; Animals ; Electrophysiology ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; metabolism ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; metabolism ; Nerve Crush ; Nerve Regeneration ; physiology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; pathology ; physiology

OBJECTIVE: To explore the general reproductive toxicity in mice exposed to low-level ozone. METHODS: Low-level (0.09 approximately 0.18 mg/m3) ozone was created by 15 W ultraviolet light. The mice in 3 experimental groups and a control group were fed in low-level ozone environment or normal environment, respectively, and then the mating experiment was conducted. The pregnancy rate and the weight variations of the female mice were observed. The weight of the live fetuses was observed, and the appearance, bone and internal organs were checked for malformation. RESULTS: There were no significant differences in any indexes between the experimental groups and the control group. CONCLUSION Low-level ozone created by 15 W ultraviolet light may not have reproductive toxicity in mice.
Animals ; Dose-Response Relationship, Drug ; Female ; Fertility ; drug effects ; Inhalation Exposure ; adverse effects ; Male ; Mice ; No-Observed-Adverse-Effect Level ; Ozone ; toxicity ; Random Allocation ; Reproduction ; drug effects ; Ultraviolet Rays

Objective:To investigate the effect of IL-17A on the differentiation and maturation of murine bone marrow-derived dendritic cells( BMDCs ) . Methods: Murine bone marrow cells were isolated and cultured in RPMI1640 complete medium in the presence of GM-CSF(20 ng/ml) for 8 days to induce differentiation of murine bone marrow cells to DC progenitors. Then these cells were treated with LPS(1 μg/ml) for 36 h which polarized immature DCs into mature DCs. Different concentrations of rmIL-17A(10 or 100 ng/ml) was added to the culture medium at different stages of BMDC differentiation and maturation. Co-stimulatory molecules expression on BMDC were analyzed by flow cytometry,and the culture supernatants were analyzed for IL-12p40 and IL-10 level by ELISA. Results:rmIL-17 could promote co-stimulatory molecules( CD40,CD80,CD86 and MHCⅡ) expression on BMDCs in a does-dependent manner,especially,the expression of CD40 and MHCⅡhad a significant increase in high concentration of rmIL-17A group;rmIL-17A was added while LPS induced maturation of BMDCs. CD40,CD80,CD86 and MHCⅡexpression on BMDC increased sharply in LPS plus rmIL-17A stimulation group,besides,CD86,MHCⅡ showed a higher level expression on BMDC with the increase of con-centration of rmIL-17A. Furthermore,secretion of IL-12p40 and IL-10 increased significantly in the group of DCs treated with LPS plus low concentration of rmIL-17 compared with the group without rmIL-17(P<0. 001). However,high concentration of rmIL-17A group showed significantly higher levels of IL-12p40(P<0. 001),but there was no difference in IL-10. Conclusion:IL-17A promotes the phe-notypic development of BMDC progenitors propagated in GM-CSF and cooperate with LPS to induce BMDC differentiation and matura-tion.

Yunnan sudden death syndrome (YSDS) is an abruptly fatal disease of unknown etiology, found mostly in central or northwestern mountain area (with altitude between 1,815 and 2,225 meters) of Yunnan province from June to September every year. It occurs mostly in young female adults, with high incidences in Lisu, Yi and Miao ethnics and high familial aggregation. The clinical manifestation of YSDS is changeful and the pathological characteristic is lack of specificity. The pathogenesis may be attributed to several factors including poor hygiene and lower socioeconomic conditions, lack of Selenium or Chromium, infection of Coxsackie B virus, mushroom consumption and special geological conditions. This article reviews the epidemiologic features, clinical manifestations, pathological features, etiology and hypothesis in order to provide clues for the research of YSDS.
Adult ; China ; Death, Sudden/pathology* ; Female ; Humans ; Syndrome