Objective To further prove the multipotentiality of rabbit bone mesenchymal stem cells (MSCs) by inducing it into cardiomyocytes with 5-azacytidine. Methods MSCs from rabbit were abstracted, isolated, cultured. Then MSCs were induced by 5-azacytidine in vitro for 24 hours. After cultured for 4 weeks, MSCs’ differentiation was studied by immunohistochemistry and electron microscopy technique. Results Some of MSCs induced by 5-azacytidine expressed troponinT . Electron microscopy showed myofilaments. Conclusion MSCs which exist in bone marrow are mutipotential stem cells, and they can be induced into cardiomyocytes by 5-azacytidine in vitro.

Objective To seek for the differentially-expressed proteins in patients with kidney-yin deficiency syndrome and to screen out the specific proteins,so as to provide evidence for the establishment of objective standard of kidney deficiency syndrome of traditional Chinese medicine (TCM).Methods Five patients with typical kidney-yin deficiency syndrome and 6 normal healthy volunteers were enrolled into the study.Plasma proteins in both groups were detected by antibody chip,and then the plasma proteins profile was compared and analyzed.Results A total of 25 differentially-expressed proteins between kidney-yin deficiency group and normal control group were found,of which 2 were up-regulated and 23 were down-regulated.Conclusion The differentially-expressed proteins in patients with kidney-yin deficiency syndrome are mainly related to immune disorder,protein biosynthesis,metabolism,oxidative stress,cell apoptosis,signal transduction,and so on.

It was the first time that the complotypes of 84 chromosomes from Han nationality healthyChinese families investigated and analyzed in Guangdong are a with the methods adopted fromthe international complement reference laboratory.The results showed 18 various complolypeswere identified.The precedent complotypes are SC31,SC42,SC32 and SC41.There were 7 com-potypes of them existing the linkage disequilibria.The highest allotypes gene frequencies of thefour loci of complotypes were respectively as follows,BF S for BF alleles,C2C for C2 alleles,C4A3 and C4A4 for C4A alleles,C4B2 and C4B1 for C4B alleles.The C4AQO allele for C4A lo-cus was 4.8% and 1.2%C4BQO alleles for C4B locus.The frequencies of major complolypesfrom the data showed the similarities among Mongolian,Caucasian,Japanese and south easternAsia populations.The significance of the different frepuencies between the complotypes has beendiscussed.

SCID mouse-human leukemia model is an important and useful tool for study on proliferation, differentiation and modulation of leukemic cells. In this article, the establishment of the model, advances in research and application in studies of pathogenesis, cell biology, clinical diagnosis, therapy and assessment of prognosis of leukemia patients are reviewed. The limitations of the model are also commented.
Animals ; Disease Models, Animal ; Humans ; Leukemia ; therapy ; Mice ; Mice, SCID ; Research Design

Objective To evaluate the efficacy of superficial temporal artery(STA)pressure-guided selective cerebral perfusion(SCP)during deep hypothermic circulatory arrest(DHCA)in patients undergoing aortic arch surgery.Methods Ninety-six patients of both sexes,aged 35-64 yr,with body mass index of 19-23kg/m2,of American Society of Anesthesiologists physical status Ⅲ or Ⅳ,undergoing aortic arch surgery,were divided into STA pressure group(group A)and clinical experience group(group B)using a random number table,with 48 patients in each group.In group A,STA catheterization was performed after tracheal intubation,and arterial pressure was monitored.SCP flow was adjusted to maintain the target value of STA pressure between 30 and 40mmHg during DHCA in group A.SCP flow rate was set at 5-10ml·kg-1·min-1 according to clinical experience in group B.The volume of fluid perfused during SCP,emergence time,extubation time and duration of intensive care unit stay were recorded.Neurological function was evaluated during length of hospitalization after surgery,and the development of permanent and transient neurological dysfunction and mortality in hospital were recorded.Results Compared with group B,the volume of fluid perfused during SCP was significantly decreased,the emergence time,extubation time and duration of intensive care unit stay were shortened,the incidence of permanent and transient neurological dysfunction(2% and 4%,respectively)was decreased(P < 0.05),and no significant change was found in the mortality rate in hospital in group A(P>0.05).Conclusion Maintaining STA pressure at 30-40mmHg is a reliable method for guiding SCP during DHCA in patients undergoing aortic arch surgery.

BACKGROUNDStent thrombosis is one of severe complications after sirolimus-eluting stent implantation. Rapamycin (sirolimus) promotes arterial thrombosis in in vivo studies. However, the underlying molecular and transcriptional mechanisms of this adverse effect have not been thoroughly investigated. This study was designed to examine the effects of rapamycin on the expression of the gene, Kruppel-like factor 2 (KLF2), and its transcriptional targets in mice.

METHODSMice were randomly divided into four groups: the control group (intraperitoneal injection with 2.5% of dimethyl sulfoxide (DMSO) only), rapamycin group (intraperitoneal injection with 2 mg/kg of rapamycin only), Ad-LacZ + rapamycin group (carotid arterial incubation with Ad-LacZ plus intraperitoneal injection with 2 mg/kg of rapamycin 10 days later), and Ad-KLF2 + rapamycin group (carotid arterial incubation with Ad-KLF2 plus intraperitoneal injection with 2 mg/kg rapamycin 10 days later). The carotid arterial thrombosis formation was induced by FeCl3 and the time of arterial thrombosis was determined. Finally, the RNA and protein of carotid arteries were extracted for KLF2, tissue factor (TF), plasminogen activator inhibitor-1 (PAI-1), endothelial nitric oxide synthase (eNOS), thrombomodulin (TM) mRNA and protein analysis.

RESULTSCompared with controls, treatment with rapamycin inhibited KLF2, eNOS and TM mRNA and protein expression, and enhanced TF and PAI-1 mRNA and protein expression, and shortened time to thrombotic occlusion from (1282 ± 347) seconds to (715 ± 120) seconds (P < 0.01) in vivo. Overexpression of KLF2 strongly reversed rapamycin-induced effects on KLF2, eNOS, TM, TF and PAI-1 expression. KLF2 overexpression increased the time to thrombotic occlusion to control levels in vivo.

CONCLUSIONSRapamycin induced an inhibition of KLF2 expression and an imbalance of anti- and pro-thrombotic gene expression, which promoted arterial thrombosis in vivo. Overexpression of KLF2 increased KLF2 expression and reversed time to thrombosis in vivo.

Animals ; Carotid Arteries ; metabolism ; Drug-Eluting Stents ; adverse effects ; Kruppel-Like Transcription Factors ; analysis ; genetics ; physiology ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; physiology ; Plasminogen Activator Inhibitor 1 ; physiology ; Sirolimus ; pharmacology ; Thrombomodulin ; physiology ; Thrombosis ; chemically induced

In vivo tumor imaging technique method based on bioluminescence principle was established to evaluate the anti-tumor effect of paclitaxel mixed micelle (PMM). MDA-MB-231 tumor cells with luciferase reporter vectors were firstly implanted into nude mice, and subsequently the luciferase substrate was regularly injected during intraperitoneal administration of PMM. Then the tumor size, growth and the intensity of light signals were monitored with in vivo imaging technique. The method of luciferase tumor in vivo imaging could be real-time, reliable and exact in labeling and reflecting the growth of tumors, and the observed results were consistent with that by conventional method, so it would be a feasible approach to study anti-tumor effect of drugs. The anti-tumor effect of paclitaxel mixed micelle was observed by this method, and the results showed that this formulation could inhibit growth of tumor, and the anti-tumor rate of it was about 85%.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; Cell Line, Tumor ; Female ; Humans ; Luminescent Measurements ; Male ; Melanoma, Experimental ; drug therapy ; pathology ; Mice ; Mice, Inbred C57BL ; Mice, Nude ; Micelles ; Neoplasm Transplantation ; Paclitaxel ; administration & dosage ; pharmacology ; therapeutic use ; Particle Size ; Tumor Burden ; drug effects

OBJECTIVETo compare therapeutic effects of acupuncture at Xuanzhong (GB 39) combined with Chinese herbs pyrogenic dressing therapy and common acupuncture on calcaneus spur.

METHODSA single-blind, randomized and controlled trial was adopted. Sixty-six cases were randomly divided into an observation group (n=34) and a common acupuncture group (n=32). The observation group was treated with acupuncture at Xuanzhong (GB 39) combined with Chinese herbs pyrogenic dressing therapy and the common acupuncture group with common acupuncture, Yanglingquan (GB 34), Kunlun (BL 60) etc. selected. The markedly effective rate and the changes of heel pain scores, heel swelling scores, heel burning sensation scores, and walking function scores were compared between the two groups before and after treatment.

RESULTSThe markedly effective rate of 64.7% (22/34) in the observation group was higher than 37.5% (12/32) in the common acupuncture group (P<0.05). After treatment, all the scores in the two groups were significantly improved as compared with before treatment (all P<0.05), and the observation group was better than the common acupuncture group (all P<0.05).

CONCLUSIONThe therapeutic effect of acupuncture at Xuanzhong (GB 39) combined with Chinese herbs pyrogenic dressing therapy on calcaneus spur is superior to that of common acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Calcaneus ; drug effects ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Middle Aged ; Osteophyte ; drug therapy ; therapy ; Treatment Outcome

OBJECTIVETo study the expression of connexin 43 (Cx43) in various stages of oral carcinogenesis and explore the relation between Cx43 and oral mucous carcinogenesis.

METHODS4-nitroquinoline-1-oxide (4NQO) was used for inducing oral carcinogenesis in SD rats. Tissue samples were obtained from various stages of the disease including normal oral mucosa, precancerous lesions and oral squamous cell carcinoma. Immunohistochemical method was used to determine the expression of Cx43 in various stages of oral carcinogenesis.

RESULTSIn the normal rat lingual mucosa, immunohistochemical staining of Cx43 protein was mainly found in the cell membrane, weakly positive in the basal cell layer, increased in stratum spinosum and stratum granulosum, but was negative in the stratum corneum of normal epithelia. Compared with normal epithelia, was significantly decreased in dysplastic and cancerous oral epithelia the staining. The positive rates of Cx43 were respectively 100.00% (10/10), 85.71% (12/14), 66.67% (8/12), 40.00% (4/10), and 33.33% (4/12) in tongue carcinogenesis (in normal, mild, moderate and severe dysplasia, and squamous cell carcinoma tissues). The differences were statistically significant (P<0.05).

CONCLUSIONExpression level of Cx43 protein was dramatically decreased with the development of rat tongue carcinoma induced by 4NQO, suggesting that abnormal expression of Cx43 protein is involved in oral mucosa carcinogenesis. Decreased Cx43 expression is an early sign of oral mucosa carcinogenesis.

4-Nitroquinoline-1-oxide ; toxicity ; Animals ; Carcinoma, Squamous Cell ; chemically induced ; chemistry ; etiology ; Connexin 43 ; analysis ; genetics ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Tongue Neoplasms ; chemically induced ; chemistry ; etiology

OBJECTIVETo express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.

METHODSHA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.

RESULTSWe successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.

CONCLUSIONWe expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.

Animals ; Blotting, Western ; Cell Line ; Drosophila ; Female ; Gene Expression ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; chemistry ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; metabolism ; Influenza, Human ; virology ; Mice ; Mice, Inbred BALB C ; Solubility