Objective To investigate the clinical effect of the blood transfusion with equal ratio component in severe multiple in‐juries with acute traumatic coagulopathy(ATC) .Methods Thirty‐eight patients who had severe multiple injuries with ATC were divided randomly into control group and treatment group .Control group was treated with the different ratio packed red blood cells (PRBC)and fresh frozen plasma(FFP) ,while treatment group received the equal ratio PRBC and FFP .Hemoglobin(HB) ,pro‐thrombin time(PT) ,international normalized ratio(INR) ,fibrinogen(FIB)were measured on the 1st ,2nd ,3rd day after admission . The total amount of PRBC during these 3 days ,the days of hospitalization in ICU ,the corrected rate of shock ,the 28‐day mortality were compared between groups .Results Compared with the control group ,the levels of PT ,INR and FIB of treatment group on the 2nd ,3rd day after admission were better(P<0 .05) .The total amount of PRBC[(18 .5 ± 6 .3)U]during these 3 days ,the days of hospitalization in ICU [(5 .9 ± 4 .3)d] in treatment group were less than those in the control group [(25 .9 ± 7 .8)U ,(10 .5 ± 7 .6)d] (P<0 .05) ,while the corrected rate of shock(85 .0% )in treatment group was higher than that of the control group(44 .4% ) .The 28‐day mortality(10 .0% )in treatment group was lower than that of the control group(27 .8% )(P<0 .05) .Conclusion The blood transfusion with equal ratio component in severe multiple injuries with ATC could not only improve blood clotting index ,reduce the total amount of PRBC and the time in ICU ,but also increase the corrected rate of shock and decrease the 28‐day mortality .

Objective To establish the murine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation (HSCT). Methods T cell-depleted (TCD) BM cells from allogeneic C57BL/6 mice were harvested and prepared from the marrow of the femurs and tibias. Age of 4 to 6 weeks’ BALB/c mice received 900 cGy total body irradiation ( TBI) from a 137Cs source. Two to four hours later,the mice were injected intravenously ( i. v. ) 5 × 106 TCD-BM cells with 1. 25 × 106 or 2. 5 × 106 or 5 × 106 splenic cells from allogeneic C57BL/6 mice,and respectively divided them into experimental group 1,experimental group 2 and experimental group 3. TCD-BM only was used as the negative control group of GVHD. Ten mice were used for each group. The establishment of aGVHD model was evaluated with a clinical GVHD scoring system,survival rate and target-organ damage. Results On 7 to 14 days after transplanta-tion,the typical clinical GVHD manifestation of severe diarrhea,hogback,activity reduced and ruffling were observed in experimental group 2. Furthermore,the aGVHD target organs of colon,lung and liver were harvested and made histological paraffin sections,then the obviously path-ological tissue damages of GVHD were detected under microscope. And the survival rate was lowed down to 0 on 45 days after transplantation in experimental group 2. On the contrary,no obviously clinical manifestation of aGVHD were observed in the control group,experimental group 1 and group 3. On 60 days later, the survival rate was 80% in experimental group 1 and 100% in control group. However,no mice was sur-vived on 10 days later in group 3. Conclusion BALB/c mice aGVHD model after allogeneic HSCT is successfully established by injecting i. v. 5 × 106 TCD-BM cells with 2. 5 × 106 splenic cells from allogeneic C57BL/6 mice.

0.05). Cross-reactivity was seen in 1 case of clonorchiasis sinensis by both the methods. Conclusion Microwave ELISA has the advantages of high sensitivity, specificity and rapidity.

Objective To study whether the organs besides digestive system of musca domestica Ⅲ instar larvae have the capability of produceing musca β-glucosidase.Methods Tissues of malpighian tubules,trachea,epiploon and body wall of musca domestica Ⅲ instar lar-vae were dissected under anatomic microscope,and the expression of β-glucosidase gene in these dissected tissues were detected by reverse transcription PCR.And the tissue localization of β-glucosidase mRNA was further identified by in situ hybridization.Moreover,anti-cellulase was used to determinate the tissue distribution with immunohistochemical staining.The relative mRNA expression levels of musca domesticaβ-glucosidase gene in these organs were tested by real-time quantitative PCR.Results The reverse transcription PCR showed that the ampli-fication products of β-glucosidase gene were observed in tissues of malpighian tubules,trachea and body wall.β-glucosidase mRNA was shown in the epithelium cells of malpighian tubules,trachea and body wall by in situ hybridization,and it was almost the same in the results of im-munohistochemical staining.The real-time quantitative PCR showed that the relative expression quantity of β-glucosidase gene in malpighian tubules and body wall were higher than that in foregut,while it was lower in itrachea than that in foregut.And it was of statistical difference in gene expression level of β-glucosidase among these organs (P <0.05).Conclusion Malpighian tubules,trachea and body wall of musca domestica Ⅲ instar larvae have the function of secreting β-glucosidase.Combining with the characteristics of secreting β-glucosidase in most organs of digestive system,it may provide a new biological method for the prevention and treatment of human diseases transmitted by musca domestica with the use of taget gene β-glucosidase.

Objective To understand the pattern of and influencing factors on leisure time physical exercises among adults in Zhejiang. Method Totally 17 437 residents aged 18 years and over selected by stratified cluster random sampling in Zhejiang in 2010 were investigated. Result Overall 21.82%(95%CI:17.12%-27.38%) of adults took part in leisure time physical exercises in 2010. The proportion was higher in urban (29.04%,95%CI:23.46%-35.33%) than in rural (18.81%,95%CI:12.89%-26.61%) (χ2=222.06, P<0.05) areas, and also higher in females (22.91%,95%CI:17.66%-29.15%) than in males (20.56%,95%CI:16.39%-25.48%) (χ2=13.94, P<0.05). Adults aged 18-24 and 55-74 years were more likely to take apart in physical exercises (χ2=266.73, P<0.05), and the lowest proportion was found among farmers (5.33%,95%CI:3.99%-7.11%) (χ2=2 078.40, P<0.05). These proportions both increased along with education level and family income increase (χ2=444.87, P<0.05;χ2=332.20, P<0.05). Overall 83.04%and 90.08%of physical exercisers took part in exercises at least 3 times per week and 30 minutes per time, only 30.67% of those reached moderate or vigorous intensity, and overall 5.38% (95%CI: 3.85%-7.48%) of adults took part in regular leisure time physical exercises in 2010. Multiple logistic regression showed that age, education level, occupation and chronic disease history have influence on regular physical exercises. Conclusion The leisure time physical exercises level was generally low among adults in Zhejiang. Young and middle-aged adults with less education and low income especially farmers should be put on emphasis.

Objective To isolate and culture the rat limbal stem cells ( LSCs) in vitro,and investigate briefly their capacity for transdif-ferentiation into neural stem cells ( NSCs) with cytokine EGF,bFGF and RA.Methods LSCs derived from rats were cultured and identified by immunohistochemistry in vitro.LSCs were induced to differentiate into NSCs in the presence of EGF (20 ng/mL) ,bFGF (10 ng/mL) and with or without of RA(group 1 or group 2)(25 ng/mL) for seven days.Cultures without factors were used as control group.Then the neural marker Nestin of the coultured cells were measured by immunohistology staining.Furthermore,the positive cell rate was counted under micro-scope between the 2 groups and analyzed by statistical software.Results It showed that P63 was positive in LSCs.Nestin in both of the dif-ferentiation groups was positive at the rate of (77.01 ±6.32)%and (84.01 ±5.43)%respectively,of which the second group was higher than the first one (P<0.05).However,it was negative in the control group.A band of Nestin protein from cells was detected by western blot assay.Conclusion LSCs are successfully isolated and cultured in vitro.LSCs could be induced to differentiate into NSCs in the presence of EGF and bFGF.Moreover,the differentiation capability is enhanced in the condition of RA.

Objective To investgate the effect of nervous growth factor(NGF) on the proliferation of the limbal stem cells(LSCs) in vitro,and the relationship bewteen expression of its receptors and cell proliferation.Methods After primary cultured,LSCs were divided into the control group and the NGF group.Selected cells cultured of 1 d,3 d and 5 d in the two groups and examined the expression of p63,TrkA,p75 with immunohistochemistry.Results The average gray scale values of expression of p63,TrkA and p75 at 1 d,3 d and 5 d in NGF group were significant decreased compared with the corresponding data in the control group(P<0.05).Pearson's correlations analysis showed that the average gray scale values of expression of TrkA and p63 were of statistically significant differences(P<0.05).Conclusion These results highlight that NGF could maintain the stem cell properties of LSCs.LSCs could exepress the NGF receptors of TrkA and p75,and the expression of TrkA showed a correlation with LSCs proliferation.

0.05).?2-MG mass concentration was significantly increased in the cerebrospinal fluid(P

Based on the transcriptome database of Salvia miltiorrhiza, specific primers were designed to clone a full-length cDNA of ent-kaurene oxidase synthase (SmKOL) using the RACE strategy. ORF Finder was used to find the open reading frame of SmKOL cDNA, and ClustalW has been performed to analysis the multiple amino acid sequence alignment. Phylogenetic tree has been constructed using MEGA 5.1. The transcription level of SmKOL from the hairy roots induced by elicitor methyl jasmonate (MeJA) was qualifiedby real-time quantitative PCR. The full length of SmKOL cDNA was of 1 884 bp nucleotides encoding 519 amino acids. The molecular weight of the SmKOL protein was about 58.88 kDa with isoelectric point (pI) of 7.62. Results of real-time quantitative PCR analyses indicated that the level of SmKOL mRNA expression in hairy roots was increased by elicitor oMeJA, and reached maximum in 36 h. The full-length cDNA of SmKOL was cloned from S. miltiorrhiza hairy root, which provides a target gene for further studies of its function, gibberellin biosynthesis and regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; methods ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Protein Structure, Tertiary ; Salvia miltiorrhiza ; enzymology

OBJECTIVE: To establish spinal muscular atrophy (SMA) cell model by blocking the expression of SMN1 gene with shRNA. METHODS: The recombinant SMN1 shRNA expression vector was constructed. SMA cell model was established by human mesenchymal stem cells(hMSCs) that the vector was transfected into were differentiated to neuron like cells (NLCs).At the same time the control groups were established that the shRNA-0 vector was transfected into and no vector was transfected into. The expression of fl-SMN and delta7-SMN mRNA was observed by RT-PCR analysis. The expression of fl-SMN protein was detected by Western blot. RESULTS: The cells of all the groups were neuron like cells after being differentiated and the protein expression of NSE and NF was positive. The expression of fl-SMN and delta7-SMN mRNA and protein of NLCs in each group was upregulated (P<0.05), but the expression of delta7-SMN mRNA and protein in SMA model group was lower than that in the control group (P<0.05). The expression of delta7-SMN mRNA between the groups had no statistical difference (P>0.05). CONCLUSION The NLCs, which recombinant SMN1 shRNA expression vector was transfected into, can be regarded as SMA cell model.
Cell Differentiation ; Cells, Cultured ; Genetic Vectors ; genetics ; Humans ; Mesenchymal Stem Cells ; cytology ; metabolism ; Models, Biological ; Neurons ; cytology ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Spinal Muscular Atrophies of Childhood ; genetics ; pathology ; Survival of Motor Neuron 1 Protein ; genetics ; metabolism ; Transfection