BACKGROUND:There are a lot of adipose-derived stem cel s in the vascular stroma. These cel s are shown to play a very important role in the fat granule transplantation.
OBJECTIVE:To explore the role of adipose-derived stem cel s in the fat granule transplantation.
METHODS:Normal adipose tissues were obtained from 10 male BALB/C mice, SPF grade. Adipose-derived stem cel s and fat granules were extracted from the abdominal fat tissues. Another 24 nude mice acted as recipients and were assigned into control, fat granule transplantation or mixed transplantation (adipose-derived stem cel s+fat granules) groups. In the latter two groups, fat granule suspension and suspension of fat granules and adipose-derived stem cel s were injected into the shoulder of rats, respectively. In the control group, the same volume of cel medium was injected. Four weeks later, separated plasma and grafts were taken out for indicator measurement.
RESULTS AND CONCLUSION:Compared with the fat granule transplantation group, the mixed transplantation could remarkably increase the weight of grafts, while reduce the absorption of grafted fat tissues (P<0.01). After transplantation, the highest level of vascular endothelial growth factor in the plasma was obtained in the mixed transplantation group fol owed by fat granule transplantation group and control group (P<0.01). Level of basic fibroblast growth factor and microvessel density were significantly higher in the mixed transplantation group than the fat granule transplantation group (P<0.01). Better cel morphology and higher number of fat droplets were found in the mixed transplantation group compared with the fat granule transplantation group. Al these results indicate that adipose-derived stem cel transplantation can remarkably promote the expression of basic fibroblast growth factor, improve graft microcirculation, and improve morphology and function of fat granules.

Purpose:The image information and technique of positron emitter nuclei generated by high energy X-ray photonuclear reactions with body tissues from MM50 is studied.It is explored to verify for dose delivery and location monitoring in tumor target after high energy photon radiation therapy with the image information.Materials and Methods:The technique is based on the photonuclear reactio-as in body tissues elemental composition ~(12)C,~(16)O and ~(14)N with high energy X-Ray,energies well above 20 MeV,resulting primarily in ~(11)C and ~(15)O but also ~(13)N.The induced positron activity distributions were scanned off-line in a PET/CT after irradiation.The activity distributions and position may be used to verify for dose delivery and location in tumor target.These radiations are similar to RT in three fields with some different dose delivery from TPS.The phantom began to be scanned off-line in a PET/CT a couple of minutes after irradiation.The scanning time is respectively 20 minutes and(2～5)mniutes for ~(12)C and ~(16)O.The levels of the beam energy are 10MV,25MV,50 MV.The extent of dose is 1.0 Gy～10.0 Gy.Since measured PET images change with time post irradiation,as a result of the different decay tim-es of the radionuclides,the signals from activated ~(12)C,~(16)O within the irradiated volume could be separated from each other.Most informationis obrained from the carbon and oxygen radionuclide's which the most abundant elements are in soft tissue.A brain phantom Rlade oneself was irradiated with high energy X-ray beams from IBA MM50.The 3D radiation treatment planning system,Nucletron-TPP 3.2,was used to calculate the delivered dose distributions.The phantom was directly simulated in the Nucletron-TPP after CT scan.PET/CT is fro-m GE Discovery LS Ⅱ.Resuits:It was confirmed that no activity was detected at 10 Mv X-ray energy,which was far below the energy threshold for photonuclear reactions.Totally 25 MV X-ray beams can produce photonuclear reactions and get to activity distributions images in PET/CT.It need to greater dose in order to good images information.For 50 MV X-ray beams,2 Gy-3 Gy dose,a normal RT dose,can get to activity distributions images.Conclusions:It was concluded that the PET-CT image acquired from the activity of the ~(15)O and ~(11)C positron emitter nuclei might provide the area and dose distribution information of 50 MV X-ray irradiation in a phantom.It can verify the in vivo dose delivery and location in tumor target after high energy X-ray RT.

Accidents, Occupational ; prevention & control ; Carbon Dioxide ; poisoning ; Facility Design and Construction ; Humans

To investigate the relationship between nm23-H1 gene and human chronic myeloblastic leukemia we designed siRNAs which target nm23-H1 gene. According to the principles of designing siRNA, we selected three siRNAs and transfected them into K562 cells by lipofectamine2000. The expression levels of nm23-H1 mRNA were detected by reverse transcriptase polymerase chain reaction after transfection for 24 hours. The expression levels of nm23-H1 protein were assayed by immunocytochemical method after transfection for 48 hours. And after transfection for 24, 48 and 72 hours, cell proliferation was determined by MTT method. Among the three siRNAs, siNM526 can effectively inhibit the expression of nm23-H1 on mRNA and protein levels. The growth of K562 cells was suppressed after transfection of siNM526. These results suggest that low expression level of nm23-H1 in K562 cells inhibited cell proliferation, namely reduced malignant degree of them. Therefore nm23-H1 gene might be a potential target of leukemia treatment.
Cell Proliferation ; Humans ; K562 Cells ; Leukemia, Erythroblastic, Acute ; genetics ; pathology ; NM23 Nucleoside Diphosphate Kinases ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

The aim of the present study was to assess the ototoxicity of kanamycin sulfate (KM) in adult rats and its underlying mechanism. Forty male Sprague-Dawley rats (6-7 weeks old) were randomly divided into the experimental group and the control group. The animals in the experimental group were injected subcutaneously with KM (500 mg/kg per day) for two weeks, and the control group received equal volume of normal saline. To assess the ototoxicity of KM, the auditory brainstem response (ABR) was recorded to monitor the changes in hearing thresholds, and the density of spiral ganglion cells (SGCs) and morphology of cochlea were observed using surface preparations and frozen sections of cochlea. The results showed that the hearing threshold of rats in the experimental group was elevated by more than 60 dB across all the frequencies two weeks after the first administration of KM. And in the experimental group, the density of SGCs became lower, and organ of Corti suffered loss of hair cells. The loss of outer hair cells (OHCs) was more severe than that of inner hair cells (IHCs), correlated with the density decrease of SGCs. We conclude that the ototoxicity of KM in the adult rats was apparent and the underlying mechanism is associated with the loss of SGCs and hair cells.
Animals ; Cochlea ; drug effects ; pathology ; Evoked Potentials, Auditory, Brain Stem ; drug effects ; Hair Cells, Auditory, Outer ; cytology ; drug effects ; pathology ; Hearing Loss ; chemically induced ; physiopathology ; Kanamycin ; toxicity ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spiral Ganglion ; pathology ; physiology ; ultrastructure

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) efficiently in pilot scale, cells of rhNDPK-A producing E. coli were homogenized by high pressure under 4 degrees C, 950 Pa. The insoluble debris was removed by microfiltration and the soluble portion was concentrated by ultrafiltration. The resulted crude sample was loaded on DEAE-sepharose Fast Flow. The target fraction was collected and then load on Cibacron Blue 3GA Sepharose CL-4B. Eluted with buffer containing ATP from the AC column, rhNDPK-A was polished with ultrafiltration. The results showed that after homogenized 2 rounds, 1500g cells of E. coli brought crude sample containing 47.6g NDPK-A. Treated with microfiltration and ultrafiltration, 27.3g of NDPK-A were recovered from this bacteria homogenate. After 2-step purification with column chromatography and then polished with ultrafiltration, 17.2 g rhNDPK-A were collected with purity of 96.3%. The recovery of the whole purification process was 36.2%, and the productivity of rhNDPK-A was 1.15 g per 100 g wet cells. Comparing the recovery of each purification step, it was found that the recovery of polish is higher than that of affinity chromatography, which is higher than that of ion exchange chromatography. The limit step was the process of sample pretreatment among the 4 purification steps. Combine with the fermentation results reported before, it was deduced that the productivity of rhNDPK-A was 510 mg/L. In conclusion, an easily controlled purification condition with high yield provides material for the translation researches of NDPK; In addition, it was suggested the crucial step determine the recovery of non-secretive recombinant proteins might be the process of sample pretreatment, not be the process of column chromatography.
Chromatography, Affinity ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Humans ; NM23 Nucleoside Diphosphate Kinases ; genetics ; metabolism ; Pilot Projects ; Recombinant Proteins ; isolation & purification ; metabolism ; Ultrafiltration

Objective To evaluate the level of regional public health development in Zhejiang Province,and to learn the development problems in order to provide suggestions for developing public health strategies.Methods The index system established for evaluation included 30 indexes of public health in 4 aspects,namely the health level of residents,the allocation of public health resource,the level of public health service and public health security.Based on the data in 2010,the comprehensive development index was calculated to evaluate the level and speed of public health development. Results The comprehensive development index of Zhejiang Province was 127. 31 in 2012,which was 9. 97 higher than thatof2011.Thespeedofdevelopmentwas108.50%,andthelevelofpublichealthdevelopmentinZhejiangProvincehas increased continuously.Among 1 1 cities,the highest and lowest comprehensive development index of public health development was 146. 35 and 101. 55 respectively.The fastest and slowest speed of development was 122. 76% and 104. 95% respectively.Conclusion The public health in Zhejiang Province kept the good momentum of development,but the unbalanced development still existed among different regions.

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) and determine its physical and chemical characters, recombinant NDPK-A producing E. coli was cultured in 80L fermentor under high cell density culture (HCDC) conditions. The harvested cells were treated with high pressure to break the cell up, tangential-flow microfiltration to remove the bacteria debris and ultrafiltration to concentrate the filtered solution containing target protein. The crude NDPK-A was purified by ion exchange chromatography with DEAE Sepharose Fast Flow, affinity chromatography with Cibarcron Blue 3GA Sepharose CL-4B and gel filtration with Sephadex G-100. The purity of rhNDPK-A was analyzed with SDS-PAGE and RP-HPLC. The Enzymatic activity was determined with RP-HPLC. The molecular weight (MW) was measured with matrix assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS). The N-terminal residue was sequenced with Edman method. The apparent molecular weight of rhNDPK-A in solution was determined with multiangle laser light-scattering method (MALS). It was found that the purity of rhNDPK-A was 97.3% with SDS-PAGE method and 99.2% with RP-HPLC method. The specific enzymatic activity was (900 +/- 100) u/mg. The molecular weight was 17017, which was 132 less than the calculated value according to the amino acid sequence of NDPK-A. The sequencing result of rhNDPK-A revealed that its N-terminal residue was Ala, which was the second residue on N-terminal of native NDPK-A. The calculated MW of N-terminal deleted rhNDPK-A was 17017, exactly equal to the experimental value. The result of apparent MW determination revealed that rhNDPK-A formed homohexamer in solution with a MW of 102kD. These results suggested that rhNDPK-A possessed character identical to its native counterpart of assembling into hexamer. Confirming the identity of rhNDPK-A to its native counterpart provided a good foundation for drug development and mechanism study of NDPK-A.
Amino Acid Sequence ; Humans ; Molecular Sequence Data ; Molecular Weight ; NM23 Nucleoside Diphosphate Kinases ; chemistry ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; isolation & purification ; Scattering, Radiation ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo construct a stable nm23-H1-knock-down cell model with K562 cell line and study its differentiation toward megakaryocyte.

METHODSEukaryotic expression vector pSilencer 4.1-CMV-sinm23 expressing siRNA targeting nm23-H1 was transfected into K562 cells with lipofectamine2000. Cells with stably nm23-H1 silence were screened out by G418. Real-time quantitative PCR, immunocytochemistry, western blot were used to confirm the nm23-H1-knock-down K562 model. Cell differentiation capacity was detected by NBT reduction assay. Surface antigen Gp IIb-IIIa (CD41) of knock-down cells treated with phorbol 12-myristate 13-acetate was analyzed by flow cytometry. Western blot was used to detect the ERK1/2 signal pathway after the stimulation of phorbol 12-myristate 13-acetate.

RESULTSEndogenous nm23-H1 was silenced by pSilencer 4.1-CMV-sinm23 and the silence efficiency was up to 75% and 70% in mRNA and protein levels respectively compared with the mock cells. Under phorbol 12-myristate 13-acetate treatment, the knock-down cells displayed a significantly increased differentiation ability toward megakaryocyte compared with control. The NBT reduction values were (0.31 +/- 0.07) and (0.23 +/- 0.05) respectively. Further results revealed that nm23-H1 gene regulating the megakaryocytic differentiation was due in part to the increased ERK1/2 phosphorylation.

CONCLUSIONSA stable nm23-H1-knock-down K562 cell model is successfully constructed. nm23-H1 involves in regulating the megakaryocytic differentiation of K562 cell line.

Cell Differentiation ; genetics ; Gene Knockdown Techniques ; Humans ; K562 Cells ; Megakaryocytes ; cytology ; NM23 Nucleoside Diphosphate Kinases ; genetics ; RNA Interference

OBJECTIVETo assess the efficacy and safety of Gastrosis No.1 compound in the treatment of functional dyspepsia with Spleen (Pi) and Stomach (Wei) deficiency-cold syndrome.

METHODSA randomized, double-blind, placebo-controlled trial was performed in 5 centers. Patients with functional dyspepsia (FD) of Spleen-deficiency and qi-stagnation syndrome (162 cases) were randomly assigned to groups given Chinese herbal medicine (CHM) Gastrosis No.1 compound or placebo in a 2:1 ratio. This trial included a 4-week treatment period and a 4-week follow-up period. The outcomes were the dyspepsia symptom scores (measured by total dyspepsia symptom scale and single dyspepsia symptom scale) and syndromes of traditional Chinese medicine score (measured by traditional Chinese medicine syndrome scale). The outcomes were noted at weeks 0, 4 and 8.

RESULTSCompared with patients in the placebo group, patients in the CHM group showed significant improvement in the dyspepsia symptom scores as rated by patients and investigators (P <0.01), and also showed improvement in syndromes of traditional Chinese medicine score (P <0.01). No serious adverse event was reported. Safety tests obtained after 4 weeks of treatment showed no abnormal values.

CONCLUSIONCHM Gastrosis No.1 compound was effective and safe in the treatment of functional dyspepsia with Spleen and Stomach deficiency-cold syndrome.

Adult ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; pharmacology ; therapeutic use ; Dyspepsia ; drug therapy ; physiopathology ; Female ; Humans ; Male ; Placebos ; Spleen ; drug effects ; physiopathology ; Stomach ; drug effects ; physiopathology ; Syndrome ; Treatment Outcome