Objective To define the correlation between mutation of DNA-LigaseⅣ gene and radiosensitivity.Methods Nasopharyngeal squamous carcinoma cell line (CNE), lung adenocarcinoma cell line (SPC-A1) and breast adenocarcinoma cell line(MCF-7) after irradiation were assessed with specific biological parameters. Polymerase chain reaction, cloning and sequencing techniques were used to determine the sequence of DNA-LigaseⅣ gene in these three cell lines. Then the impact of homologous change and mutation on hydrophicity-hydrophobicity of genic products was analyzed.Results The surviving fractions derived from irradiation were different with more radiosensitivity in the CNE cell line than in the others. In three cell lines, the homology of LigaseⅣ gene were: 99.95%,99.99%,99.98%, respectively. Some mutations including transversion and transition were detected and led to alterations in the hydrophicity-hydrophobicity function of products. Higher radiosensitivity of CNE was associated with amino-acid substitutions: 313aa His→Arg,538aa Gly→Arg,579aa Lys→Arg and 585aa Asn→Ser.Conclusion These results suggest that LigaseⅣp play an important role in the ligation of DNA double strand breaks and certain mutations bring about changes in radiosensitivity.

Objective To investigate the change in mRNA expression of hypoxia inducible factor 1? HIF 1? and vascular endothelial growth factor (VEGF) genes in the highly differentiated nasopharyngeal squamous carcinoma (CNE) cell line during hypoxia reoxygenation.Methods CNE cells were cultured in normoxic, continuous 8 hour hypoxia ,16 hour hypoxia and 16 hour hypoxia 4 hour reoxygenation conditions respectively. Expressions of HIF 1? and VEGF genes were assessed by the reverse transcription polymerase chain reaction (RT PCR) technique. Results The mRNA expression of HIF 1? gene was kept constant in the different culture conditions,whereas the mRNA expression of VEGF gene increased significantly after 8 or 16 hour hypoxia and decreased after reoxygenation for 4 hours. Conclusions Hypoxia is able to induce the mRNA expression of VEGF gene in the CNE cell, but is unable to do so in HIF 1? mRNA expression. These findings suggest that VEGF upregulation induced by hypoxia be independent HIF 1a transcription.

NDV strain ZJ1 strain , a highly virulent NDV strain, has been prevalent among the waterfowls in China mainland in the past years. Multi-basic amino acid sequence distribute in the protease cleavage site of F protein of this strain. Recombinant expressing plasmid pCI-FT, was generated by converting multi-basic amino acid sequence of 112, 115, 117 of the protease cleavage site of F_ 0 protein, to the non-basic amino acid sequence characteristic of avirulent NDV strain. The result from co-expression of mutant or parental F protein with homologous HN protein in COS-1 cells revealed that both mutant and parental F protein had fusion activity. The result from co-expression of mutant or parental F protein with homologous HN protein in CEF cells showed that the cleavage activity of mutant F protein was significantly reduced. The study built a foundation for mutagenesis of amino acid sequence of the protease cleavage site of F_ 0 protein at the full-length cDNA clone level, study on factors contributing to virulence and construction of candidate vaccine strain, and so on.

Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers.The fragments,amplified with primer Ⅰ to Ⅶ,were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain.The eukaryotic expression vector pCI-L was constructed by subcloning the fragments,amplified with the primer Ⅴ,Ⅵ and Ⅷ,into the expression vector pCI-neo.The full-length cDNA clone,pNDVZJI,with three helper plasmids,pCI-NP、pCI-P and pCI-L,were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase.After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF)flock,The NDV of ZJI strain was rescued successfully,which laid a good foundation for the further related research.

OBJECTIVETo summarize the clinical effect of avulsed skin replantation of hand and foot via vacuum sealing drainage (VSD) combing low temperature technique.

METHODSFrom March 2012 to October 2013,13 cases with avulsed skin replantation of hand foot using combined technique included 8 males and 5 females with an average age of 32 years old ranging from 18 to 62 years. The time from injury to hospital was 1 to 4 hours (2.4 hour in average). The reasons of injury included machine injury in 7 cases and rolling over by cars in 6 cases. The parts of injuried involved finger in 2 cases,back of the hand in 5 cases and dorsum of foot in 6 cases. The area of avulsed skin was 5 cm x 6 cm to 12 cm x 16 cm,tendon and bone exposure was found in 4 cases. VSD was operated in all patients and the avulsed skin was refrigerated in the temperature of -4 °C or -80 °C. After 4 days, the skin stored in the -4 °C was replanted to the wounded place in 5 cases and in 3 cases the skin was planted to the donor site of flap. The skin stored in the -80 °C was replanted in 4 cases after 7 or 8 days, 1 case after 45 days.

RESULTSOf the 13 cases, 1 case of degloved injury from lower leg to dorsal foot,the replanted skin was necrosis completely; 1 case of degloving injury with fourth finger,the skin which replanted after 45 days survived approximately 30%,cured after skin-graft many times. In the other cases, the survival area of replanted skin was more than 85%, all cured after dressing. According to the standard of skin survival area evaluation by Jia et al, 11 cases showed excellent, 1 showed medium and 1 showed inferior. There were no complication about grafted skin rupture after the skin survived in 11 patients,after 4 to 22 months follow-up, the resiliency of grafted skin showed good. Sensation recovery was measured by BMRC standard: 3 cases of S3, 5 cases of S3, 3 cases of S2.

CONCLUSIONVSD combining lower temperature technique in skin replantation provides time and space for wound preparation and treatment plan for the patients who need second surgery, especially for the large area skin degloving,this method could utilize the degloved skin efficiently, decrease the donor site area, alleviate the pain and financial burden,reduce the scar formation of donor site and impediment.

Adolescent ; Adult ; Cryopreservation ; methods ; Drainage ; instrumentation ; methods ; Female ; Foot Injuries ; surgery ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Replantation ; Skin ; injuries ; Skin Transplantation ; Young Adult

Objective: To investigate the protective effects of matrine combined with glycyrrhizic acid on chronic liver injury induced by carbon tetrachloride, and explore the protective mechanism from the points of energy metabolism and CYP enzyme.Methods: The chronic hepatic injury model of rats was induced by CCl4.The changes of activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured to observe the protective effect of the two drugs and their combination.The contents of glutamate dehydrogenase (GLDH) in serum and adenine nucleoside three phosphate (ATP), adenosine diphosphate (ADP) and adenine monophosphate (AMP) in liver tissue were determined to evaluate the regulation effect on hepatic energy metabolism and mitochondrial function.The levels of CYP1A2, CYP2E1 mRNA and protein in liver tissue were detected by real-time PCR and Western Blot to evaluate the two drugs and their combination on the regulation function of liver CYP enzyme.Results: Matrine (72.8 mg×kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease the serum activities of ALT and ALT in chronic hepatic injury model, and the combination (matrine 36.4 mg·kg-1+glycyrrhizic acid 21.7 mg·kg-1) had the most significant protective effect (P＜0.05).Matrine (72.8 mg·kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease GLDH in serum,and restore the content of ATP in liver (P＜0.05).Matrine (72.8 mg·kg-1) had no effect on the expression of CYP1A2 and CYP2E1mRNA, and glycyrrhizin (43.4 mg·kg-1) could inhibit the expression of CYP1A2, CYP2E1mRNA and protein (P＜0.05).Conclusion: Matrine combined with glycyrrhizin has obvious regulation effect on mitochondrial function and liver protective effect in chronic hepatic injury model.

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain,seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI,which contained the full-length cDNA of the NDV ZJI strain.The pNDV/ZJI,with three helper plasmids,pCIneoNP,pCIneoP and pCIneoL,were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase.After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock,an infectious NDV ZJI strain was successfully rescued.Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP.After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells,the recombinant NDV,NDV/ZJIGFP,was rescued.Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection,indicating that the GFP gene was expressed at a relatively high level.NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route.Four days post-infection,strong green fluorescence could be detected in the kidneys and tracheae,indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.

Objective To explore the effects of community synthetic nursing intervention on negative emotion of puerperium women.Methods Totals of 100 puerperium women were randomly divided into experimental group and control group,50 cases per group.The control group received conventional health education and home visits and experimental group received community synthetic nursing intervention.Score of anxiety scale ( SAS),depression scale (SDS),symptom assessment scale (SCL-90) and family function,before and after intervention,were compared between two groups.Results SAS score and SDS score in experimental group was (36.76 ± 10.62) and (37.46 ± 10.23) lower than (50.83 ± 10.85) and (49.78 ± 10.14) in control group,the difference was statistically significant (t =6.55,6.05;P ＜ 0.01 ).SCL-90 score of experimental group was lower than that of control group.The well family function in experimental group was 45 cases and 36 cases in control group,and the difference was statistically significant ( x2 =5.26,P ＜ 0.05 ).Conclusions Application of community synthetic nursing intervention in puerperal women,can eliminate harmful precipitating factor existing in person,family,society and decrease negative emotion of puerperal women.

OBJECTIVETo investigate the effect and mechanism of BSDW on the model of allergic rhinitis and the model of guinea pigs by histamine shocking in guinea pigs.

METHODUsing the model of allergic rhinitis in guinea pigs caused by 10% TDI, we observed the effect of BSDW on physiological and pathological symptoms of allergic rhinitis in guinea pigs, the effect of the levels of serum IgE and serum and nasal histamine. Using the model of guinea pigs by histamine shocking, we observed the effect of BSDW on physiological symptoms in guinea pigs.

RESULTBSDW significantly relieved the pathological symptoms of allergic rhinitis in guinea pigs, alleviated the hyperplasia of columnar epithelium, decreased the number of monocyte and eosinocyte compared with the model group. It also reduced the levels of serum IgE, and decreased the release of serum and nasal histamine. BSDW significantly prolonged the occurent time of gasping, eclampsia and death caused by shock, reduced the times of gasping in the model of guinea pigs by histamine shocking.

CONCLUSIONBSDW has significant effect against allergy. The mechanism relates to its effects of decreasing the levels of serum IgE and inhibiting the release of serum and nasal histamine.

Administration, Intranasal ; Animals ; Anti-Allergic Agents ; pharmacology ; Asarum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Guinea Pigs ; Histamine ; blood ; Immunoglobulin E ; blood ; Lamiaceae ; chemistry ; Male ; Nasal Mucosa ; immunology ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Perennial ; immunology ; Scutellaria ; chemistry ; Toluene 2,4-Diisocyanate

OBJECTIVETo investigate the mechanism of tazarotene against active psoriasis vulgaris.

METHODSA randomized, controlled trial was conducted in 43 patients with active psoriasis vulgaris, who were divided into tazarotene and control groups. Promyelocytic leukemia (PML) mRNA in active psoriatic lesions before and 14 days after tazarotene treatment was detected by in situ hybridization.

RESULTSPML mRNA expression was detected not only in the basal layer (86.96%), but also in the suprabasal layers of the epidermis in the manner of focal expression (78.26%). After tazarotene treatment, virtually no PML mRNA expression could be detected in the psoriatic lesions (8.69% in the basal layer and 4.35% in the suprabasal layers). PML mRNA expression in the control group underwent no obvious changes during the observation.

CONCLUSIONSTazarotene may inhibit abnormal proliferation of keratinocytes through down-regulating PML gene expression in active psoriatic epidermis.

Adolescent ; Adult ; Double-Blind Method ; Down-Regulation ; drug effects ; genetics ; Epidermis ; drug effects ; metabolism ; pathology ; Female ; Gene Expression ; drug effects ; Humans ; In Situ Hybridization ; Keratolytic Agents ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; Nicotinic Acids ; administration & dosage ; therapeutic use ; Nuclear Proteins ; genetics ; Promyelocytic Leukemia Protein ; Psoriasis ; drug therapy ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transcription Factors ; genetics ; Tumor Suppressor Proteins ; genetics