0.05). ConclusionsTwo methods of fibero tomy do not effect on the periodontal tissues. The PSP is more excellent than t he MSF in preventing the corrected teeth from relapsing and keeping the aest hetic feeling of the gingival shape.

Objective To explore the TCM syndrome distribution characteristics of central obesity hypertension patients; To analyze its correlation with urinary microalbumin/creatinine (MA/Cr) ratio; To provide some proof for an efficient way to control central obesity hypertension and prevent and cure its early renal injury with integrated TCM and Western medicine.Methods It was performed in a cross-sectional epidemiological study. The age, gender, height, weight, waist circumference, blood pressure, medical history and symptoms of 359 central obesity hypertension patients were collected in Shanghai. Then according to the four diagnostic methods, TCM syndromes were recorded. The urinary MA/Cr ratio, fasting blood glucose, fasting insulin and hs-CRP levels were detected.Results The urinary MA/Cr detectable rate in Shanghai among central obesity hypertension was 33.4% (120/359), men accounting for 56% and women 44%. Among central obesity hypertension patients, the HOMA-IR and hs-CRP level of urinary albumin group were significantly higher than those of normal group, with statistical significance (P<0.05). Among 359 central obesity hypertension patients, 140 people had phlegm-dampness syndrome, accounting for 39%, the largest part; 108 had liver-yang hyperactivity syndrome, accounting for 30%; 61 had yin-yang deficiency syndrome, accounting for 17%; 50 had yin-deficiency and yang-hyperactivity syndrome, accounting for 14%; the number of four TCM syndromes had statistical difference (P<0.05). Urinary MA/Cr ratio of the patients with phlegm-dampness syndrome was significantly higher than that of other three syndromes (P<0.05).Conclusion The incidence rate of early renal damage with central obesity hypertension patients is high in Shanghai area, and the early stage of renal damage is associated with insulin resistance and inflammatory reaction. Among central obesity hypertension, phlegm-dampness syndrome and liver-yang hyperactivity account for the majority, and patients with phlegm- dampness syndrome are more likely to have early kidney damage.

In the present study, the antibacterial spectrum of turmeric extract was analyzed by measuring the minimum inhibitory concentration (MIC), and the antibacterial mechanism of turmeric extract was elaborated by determining its effects on the permeability and integrity of the cytoplasmic membrane, energy metabolism, and the morphology of the tested bacteria (Bacillus subtilis) with the highest susceptibility to the extract. The results showed that turmeric extract possessed broad-spectrum antibacterial activity against Gram-positive, Gram-negative bacteria and fungi, especially against Bacillus subtilis, with the MIC of 0.5 mg·mL^-1. Turmeric extract disrupted the liposome membrane and released calcein encapsulated within it. The permeability of the bacterial cell membrane was increased, leading to leakage of intracellular K⁺, Ca²⁺and cell wall proteins, and the integrity of the cell membrane was broken down, causing leakage of intracellular proteins and polysaccharides. Scanning electron microscope observation confirmed that the bacteria treated with turmeric extract was severely deformed. Simultaneously, cellular energy metabolism was affected, resulting in a significant reduction in the intracellular ATP content and ATPase activity in bacteria. In summary, the antibacterial mode of turmeric extract is closely related to its disruption of bacterial membrane structure.

Objective To study the survival time of recombination rival in environment and inactivation ability of different disinfectant and ultraviolet radiation against virus. Methods NC membranes absorbed the recombinant adenovirus (rADV) or herpes simplex vires (rHSV) with green fluorescence protein (GFP) were laid, or immersed in various concentration of different disinfectants such as ethanol, sodium hypochlorite, lysol and geramine and then taked out them every 15 min, or exposed under ultraviolet radiation, then the NC membranes were adsorbed 1 h in cell, 37 ℃ 5 % CO248 h. The results were observed under the fluorescence microscope. Results (1) the average survival time of rHSV under environment is less than 60 min, rADV is almost up to 2 h. (2) The infection ability of rHSV and rADV was inactived 15 min by both ethanol(100%, 70% and 50%) and sodium hypochlorite (5%, 2.5% and 1.25%). (3) Two virus can be killed by 0.1% bromngeramine. (4) Both 5% and 2.5% lysol, but rADV can not lost the infection on Vero Cell until 75 min by 1.25% Lysol. (5) The rHSV was inactivated under ultraviolet radiation, but rADV was not. Conclusion The survival time of is different from beth envelope rival and the no-envelope viral under nature environment and the inactivate ability of disinfectant also is different between two model virus; Disinfectant should be choose according to virus type.

Objective To investigate the phenomenon of accidental splashes and sprays from manipulation of recombinant virus material and to measure the approximate spilled distance when recombinant virus material inadveaenfly dropped in the biosafety laboratory.Methods first,two groups owning different experience simulated the course of accidental spills and splashes by recombinant adenovirus(rADV)which expressed green fluorescence protein(GFP),the GFP signal were observed in 96 well cell plate after spills appeared;Second,the routine two heishts(75 cm and 110 cm)and capacity(1 ml,1.5 ml,4 ml and 8 ml)of virus were chose to simulate the experiment of unexpected dropping.Results First,the positive quantity of the first group owning 5 years'experience is much less than the second group owning 2 years'work experience,the former was 7 positive wells,the latter was 81 positive when they used the pipette to operation.Second,when the unclosed test tubes(1 ml,1.5ml,4 ml and 8 ml recombinant virus)inadvertently dropped,the largest spill distance Was 0.92 m、1.57 m、2.63 mand 2.68 m respectively.Conclusion The better experience is important to make sure safety when we make infectious material;the contaminated distance increased with the amount of recombinant virus material.

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.
Animals ; Antimetabolites, Antineoplastic ; metabolism ; pharmacology ; Cell Death ; drug effects ; Chick Embryo ; Cytosine Deaminase ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Flucytosine ; metabolism ; pharmacology ; Fluorouracil ; metabolism ; pharmacology ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lethal Dose 50 ; Liver Neoplasms, Experimental ; pathology ; Mice ; Newcastle disease virus ; genetics ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden ; drug effects

Animals ; Cell Transformation, Neoplastic ; Cloning, Molecular ; Hepacivirus ; genetics ; immunology ; pathogenicity ; Hepatitis C Antigens ; toxicity ; Liver Neoplasms, Experimental ; pathology ; virology ; Male ; Oncogenic Viruses ; pathogenicity ; RNA, Messenger ; toxicity ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; toxicity ; Viral Core Proteins ; toxicity ; Virion ; pathogenicity

Reproductive toxicity research takes an important place in traditional Chinese medicine pre-clinical safety evaluation. Modern reproductive toxicity experiment includes drug-related miscarriage, fetal death, teratism, and adverse effects on fertility, genital system, embryonic development and fetus, which is different from contraindicated in pregnancy in traditional Chinese medicine theory. Now the three-phases reproductive toxicity study is the method mainly applied in traditional Chinese medicine reproductive toxicity evaluation. Besides that, alternative methods of whole embryos culture and embryonic stem cell test are also used in traditional Chinese medicine embryo toxicity evaluation. This article reviews research progress and pre-clinical evaluation on reproductive toxicity of traditional Chinese medicine.
Animals ; Drug-Related Side Effects and Adverse Reactions ; Drugs, Chinese Herbal ; toxicity ; Embryonic Development ; drug effects ; Embryonic Stem Cells ; drug effects ; Female ; Humans ; Medicine, Chinese Traditional ; Pregnancy ; Reproduction ; drug effects ; Toxicity Tests

The aim of this study was to investigate the factors which affect HLA typing in 311 umbilical cord blood (UCB) samples. The HLA low resolution typing of UCB samples with misinterpreted HLA types from 311 UCB samples analyzed by PCR-SSO and PCR-SSP was performed. 7 samples difficult to determine their HLA genotype were sequenced directly and the reason leading to misinterpret HLA typing was analyzed. The results indicated that 99.4% of misinterpreted samples resulted from the restriction of HLA typing method itself and 0.6% of misinterpreted samples were suspected to be contaminated with maternal blood in UCB. It is concluded that HLA typing is mainly affected by the shortcomings of oligonucleotide probe design for PCR-SSO and lack of allele specific primers of PCR-SSP.
Alleles ; Base Sequence ; DNA Primers ; Fetal Blood ; immunology ; Genotype ; HLA Antigens ; genetics ; Histocompatibility Testing ; methods ; Humans ; Oligonucleotide Probes ; Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the biological behavior including survival and proliferation of CD34 + CD38--Lin--cells when they are cultured at single cell level.

METHODSPurified umbilical cord blood CD34 + CD38--Lin--cells were separated at single cell level in 96-well plates using flow cytometry for four groups: control group (CD34 + CD38--Lin--cell plus stem cell medium) , Shh group (CD34 + CD38--Lin--cell plus stem cell medium and Shh), BMP-4 group (CD34 + CD38--Lin--cell plus stem cell medium and BMP-4), Jagged-1 group (CD34 + CD38--Lin--cell plus stem cell medium and Jagged-1). Methylcellulose medium was used in the colony-forming experiment which was also in four groups as previously. The number of cells and colony-forming units in each well for the four groups was evaluated at different time points (day 1, 3, 7) with fluorescence microscopy counting method.

RESULTSDivision of single cell was observed to be amplified in all of these groups from day 3. And meanwhile, after 1-week culture, the survival rates for the treated groups were all higher than the control group (Jagged-1 group > BMP-4 group > Shh group > control), while the cell number in each well was also highest in the Jagged-1 group (Jagged-1 group > BMP-4 group > control). The number of wells with a cell number of zero was significantly fewer in all treated groups (especially the Jagged-1 group) than in the control group; meanwhile, the number of wells with a cell number higher than 17 was evidently higher in all the treated groups (especially the BMP-4 group) more than controls. Colony-forming units for erythroid (BFU-E), granulocyte (CFU-G), macrophage (CFU-M), and granulocyte macrophage (CFU-GM) were observed for all of these experimental groups, and there was no significant difference between the four experimental groups.

CONCLUSIONSCD34 + CD38 - Lin - cell can achieve the survival, self-renewal and proliferation when cultured at single cell level, and the adding of Shh, BMP-4, and Jagged-1 can enhance such capabilities. However, CD34 + CD38 - Lin - cell can only maintain cell totipotency in its niche.

ADP-ribosyl Cyclase 1 ; metabolism ; Antigens, CD34 ; metabolism ; Bone Morphogenetic Protein 4 ; chemistry ; Calcium-Binding Proteins ; chemistry ; Cell Culture Techniques ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Colony-Forming Units Assay ; Culture Media ; Fetal Blood ; cytology ; Hedgehog Proteins ; chemistry ; Hematopoietic Stem Cells ; cytology ; Humans ; Intercellular Signaling Peptides and Proteins ; chemistry ; Jagged-1 Protein ; Membrane Proteins ; chemistry ; Serrate-Jagged Proteins