BACKGROUND: Chinese traditional osteopathy is long in history, unique in manipulation and miraculous in therapeutic effect. But people understand it more m perception rather than in theory, more in application rather than in development. There is little research truly on the bioseience.OBJECTIVE: To probe into the macro-idea of Chinese traditional osteopathy, micro-mechanisms on characters and mathematics-physics models, aiming to provide new principles and approaches of treatment for the daily increased bone trauma, fracture and sport injury.SETTING: Physics and machine-electron college of a university, and its affiliated hospital.METHODS: Based on the natural concept of "integration between heaven and human being" and new concept of holistic medicine in Chinese traditional osteopathy, the macro-idea and characters of reduction and union of fracture are generalized from the characters of natural therapy and the biomechanical mechanisms and characters of reduction and union of fracture are summarized from the micro-reaction of bone repair and union so as to discover biomechanical mechanisms and characters of reduction and union of fracture and further to set up biomechanical models and mathematics-physics expressions during the treatment.RESULTS: Chinese traditional osteopathy envelopes macro-idea of "initiative reduction-functional union" in fracture and micro-mechanism on "stress adaptability-functional adaptability" of bone repair and union.CONCLUSION: Chinese traditional osteopathy compiles with the natural,green and non-traumatic therapy in bio-natural law of bone repair and union and supports the theme of "high thought and high skill".

Objective To establish and evaluate a method for determination of total arsenic in urine by test-tube rapid digestion hydride generation atomic fluorescence spectrometry.Methods After digestion of urine samples using graduated test-tube and graphite digestion apparatus,arsenic content in urine was determined with atomic fluorescence spectrometer.Then the test results were evaluated by using quality control measures,such as precision and accuracy experiments,and the results between different laboratories were reviewed and compared.Results The urinary arsenic was in a linear range of 0-0.300 mg/L,correlation coefficient (r) ＞ 0.999 3,detection limit was 0.000 21 mg/L,relative standard deviation (RSD) ≤4.62％ and the recoveries of standard addition were 93.9％-104.3％.The value of standard reference material measured was within the allowable range.The blind sample of the national urinary arsenic was qualified.Conclusions This method is suitable for large scale determination of urinary arsenic for its micro sample amount needed,less interference and strong practicability.The error results are in a controlled range.

With their capability to undergo unlimited self-renewal and to differentiate into all cell types in the body, induced pluripotent stem cells (iPSCs), reprogrammed from somatic cells of human patients with defined factors, hold promise for regenerative medicine because they can provide a renewable source of autologous cells for cell therapy without the concern for immune rejection. In addition, iPSCs provide a unique opportunity to model human diseases with complex genetic traits, and a panel of human diseases have been successfully modeled in vitro by patient-specific iPSCs. Despite these progresses, recent studies have raised the concern for genetic and epigenetic abnormalities of iPSCs that could contribute to the immunogenicity of some cells differentiated from iPSCs. The oncogenic potential of iPSCs is further underscored by the findings that the critical tumor suppressor p53, known as the guardian of the genome, suppresses induced pluripotency. Therefore, the clinic application of iPSCs will require the optimization of the reprogramming technology to minimize the genetic and epigenetic abnormalities associated with induced pluripotency.
Animals ; Cellular Reprogramming ; Disease Models, Animal ; Epigenesis, Genetic ; Genomic Instability ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).

METHODS(1) The interaction of STIM2 and TRPC3 was determined using the immunofluorescence technique. (2) The expressions of STIM2 and TRPC3 genes were silenced in HUVEC by transfection constructed STIM2 and TRPC3 RNA interference plasmids. The interference efficiency of STIM2, TRPC3 protein and mRNA levels were determined by Western blot and real time RT-PCR, respectively. (3) The second to fifth passage of HUVEC were divided into: STIM2-002 short hairpin RNA (STIM2-002 shRNA ) + spermine + Ca2+ group and TRPC3-004 short hairpin RNA (TRPC3-004 shRNA ) + spermine + Ca2+ group; control group (spermine + Ca2+ group) and vehicle+ spermine + Ca2+ group. The four groups of cells were incubated with CaR agonist spermine, the intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, and the production of NO was determined by DAF-FM (NO fluorescent probe) of each group in HUVEC.

RESULTS(1) Immunofluorescence technique results showed that STIM2 and TRPC3 proteinswere present in the cytoplasm of HUVEC. (2) The results of transfection constructed STIM2 and TRPC3 RNA interference plasmids demonstrated that shRNA targeted to the STIM2 and TRPC3 genes decreased STIM2 and TRPC3 mRNA levels by 88.2% and 74.0%, respectively (P < 0.05), simultaneously, the STIM2 and TRPC3 protein levels were decreased by 79.9% and 71.8%, respectively (P < 0.05). (3) Compared with spermine + Ca2+ group, the [Ca2+]i and the net NO fluorescence intensity of spermine + Ca(2+) + ShSTIM2-002 group, spermine + Ca(2+) + ShTRPC3-004 group and spermine + Ca2+ Vehicle group were not changed (P > 0.05).

CONCLUSIONSTIM2 and TRPC3 do not participate in CaR-mediated Ca2+ influx and NO production individually.

Calcium ; metabolism ; Cell Adhesion Molecules ; physiology ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; physiology ; Humans ; Nitric Oxide ; metabolism ; Stromal Interaction Molecule 2 ; TRPC Cation Channels ; physiology

OBJECTIVETo analyze the sequence polymorphisms of mitochondrial DNA HVR I and HVR II in Tibetan population in Changdu area of Tibet.

METHODSmtDNAs obtained from 97 unrelated individuals were amplified and directly sequenced.

RESULTSOne hundred and eleven variable sites were identified, including nucleotide transitions, transversions, insertions and deletions. In HVR I region (nt16024-nt16365), sixty-eight polymorphic sites and 92 haplotypes were observed, and the genetic diversity was 0.9985. In HVR II region (nt73-nt340), forty-three polymorphic sites and 91 haplotypes were detected, and the genetic diversity was 0.9882. The random match probability of HVR I and HVR II regions were 0.0120 and 0.0118, respectively. When the sequence analysis of HVR I and HVR II regions were combined, ninety-seven different haplotypes were found. The combined match probability of two unrelated persons having the same sequence was 0.0103.

CONCLUSIONThere are some unique polymorphic loci in the Changdu Tibetan population. The results suggest that there are significant difference in the genetic structure in the mitochondrial DNA D-loop region between Changdu Tibetans and other Asian populations and Caucasians. Sequence polymorphism in mitochondrial DNA HVR I and HVR II can be used as a genetic marker for forensic individual identification and genetic analysis.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; DNA, Mitochondrial ; genetics ; Ethnic Groups ; genetics ; Female ; Genetic Variation ; Genomics ; Haplotypes ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Tibet

Anoikis is a form of apoptosis induced upon cell detachment from extracellular matrix.It has been determined that acquisition of resistance to anoikis is a critical step for tumor cell metastasis.MiR-21,the most prominent oncomiR,plays an important role in tumor progression.In this study,we revealed that up-regulation of miR-21 in human esophageal adenocarcinoma (EA) is associated with lymph node metastasis and poor survival rate.Because of the established anti-apoptosis effect of miR-21,it is tempting to speculate that miR-21 might contribute to tumor metastasis by regulating anoikis,qRT-PCR analysis demonstrated that miR-21 expression in OE33/AR cells (subpopulation of human EA OE33 cells that acquired resistance to anoikis) was significantly increased.Also,transfection of miR-21 mimics provided OE33 cells resisting to anoikis.By luciferase assays,we verified that PDCD4 and PTEN were the functional targets of miR-21.In mouse model,via tail vein injection experiment,we showed that the metastasis formation of OE33 cells in vivo could be mediated by changing the miR-21 expression pattern.Taken together,our findings suggested that miR-21 was involved in the regulation of anoikis in human EA cells.Targeting miR-21 may provide a novel strategy to prevent metastasis.

OBJECTIVE：To investigate the effects and its mechanism of pseudostrychnine on the apoptosis of human colon can - cer HT- 29 cells. METHODS ：Human colon cancer HT- 29 cells were randomly divided into blank group ，pseudostrychnine low-dose，medium-dose and high-dose groups （125，250，500 μmol/L）. They were cultured with culture medium without medicine or relevant concentration of pseudostrychnine for 48 h. The cell apoptosis and mitochondrial transmembrane potential were detected by flow cytometry. Western blotting assay was employed to detect the protein expression of P 53，Caspase-3，Caspase-9，DNA re - pair enzyme from rabbit （c-PARP）and Bcl- 2. RESULTS ：Compared with blank group ，apoptotic rate of cells was increased signifi - cantly in pseudostrychnine low-dose ，medium-dose and high-dose groups ，while mitochondrial transmembrane potential was de - creased significantly （P＜0.01），in concentration-dependent manner. The protein expression levels of P 53，Caspase-3，Caspase-9 and c-PARP were increased in pseudostrychnine medium-dose and high-dose groups ，compared with blank group ；while those of Bcl-2 were decreased significantly in pseudostrychnine low-dose ，medium-dose and high-dose groups （P＜0.05 or P＜0.01）. CON - CLUSIONS：Pseudostrychnine may change mitochondrial membrane potential by up-regulating the protein expression of P 53 and down-regulating the protein expression of Bcl- 2，and activate the expression of Caspase- 3，c-PARP and Caspase- 9，so as to acti - vate endogenous mitochondrial pathway and promote the apoptosis of HT- 29 cells.

OBJECTIVETo evaluate interfering effect of several short interfering RNAs (siRNA) on HCV 5' untranslated region(5' UTR).

METHODSThe green fluorescent protein (GFP) was used as reporter gene. A fused gene of HCV-5'UTR and GFP was constructed. It was cloned into the plasmid pCDNA3.1 named as pcDNA-HCV-5'UTR-GFP. Three siRNAs were designed and transfected into HepG2 cells with pcDNA-HCV-5'UTR-GFP. The change of the fluorescence intensity of HepG2 cells was shown by fluorescence microscopy and numerically detected under 488 nm wave length by flow cytometry.

RESULTSThe fused gene of HCV-5'UTR and GFP was successfully constructed. The seven groups displayed inhibitory effects on the gene expression of GFP. The inhibition rates of siRNA A, B and C were 68.4 percent, 72.6 percent and 75.6 percent, respectively. The inhibitory rates of siRN A + B, siRN B +C and siRN A +C were 91.8 percent, 87.2 percent and 92.4 percent, respectively. The inhibitory rates of siRN A+B +C was the highest, up to 95.7 percent.

CONCLUSIONThese siRNAs could inhibit expression of HCV 5'UTR gene, the inhibitory effect of combined siRNA was better than that of single siRNAs.

5' Untranslated Regions ; genetics ; Green Fluorescent Proteins ; genetics ; Hepacivirus ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo compare the protein difference between B. henselae Houston and B. henselae Marseille by two-dimensional gel electrophoresis.

METHODProtein samples were prepared by vorterx, ultrasonic treatment, and centrifugation. Protein concentrations were determined by Bradford method. Protein difference was compared by the first IEF and the second SDS-polyacrylamide gel electrophoresis.

RESULTSProtein concentrations in samples of Bartonella henselae Houston and Bartonella henselae Marseille were 2.117 microg/microL and 2.200 microg/microL respectively. Sample protein of 40 microg for IPG strips loading was perfect. The results of 2-DE in pH 4 to 7 IPG strips showed that the total protein spots of Bartonella henselae Houston and Bartonella henselae Marseille were 375 and 379 respectively, 95% of the spots were the same between the two strains of Bartonella henselae.

CONCLUSIONThe procedure of 2-DE may prove successful for the proteomic analysis of Bartonella henselae. Bartonella henselae Houston and Bartonella henselae Marseille are different genotypes.

Bacterial Proteins ; analysis ; metabolism ; Bartonella henselae ; classification ; genetics ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Genotype ; Proteomics

The actions of four organic acids in Radix isatidis, a traditional Chinese medicinal (TCM) herb, on Escherichia coli, Staphylococcus aureus and Shigella dysenteriae growth were investigated by microcalorimetry. The four organic acids were syringic acid, 2-amino-benzoic acid, salicylic acid and benzoic acid. The power-time curves of Escherichia coli, Staphylococcus aureus and Shigella dysenteriae growth with and without organic acids were acquired, meanwhile the extent and duration of inhibitory effects on the metabolism were evaluated by growth rate constants (k1, k2), maximum heat-output[0] power (P(m)) and peak time (t(m)). The inhibitory activity varied with different drugs. The sequences of anti-microbial activity of the four organic acids on Escherichia coli, Staphylococcus aureus and Shigella dysenteriae were all: syringic acid > 2-amino-benzoic acid > salicylic acid > benzoic acid. And benzoic acid promoted the growth of Staphylococcus aureus and Shigella dysenteriae. This study provides a basis for the further study on Radix Isatidis.
Anti-Bacterial Agents ; isolation & purification ; pharmacology ; Benzoic Acid ; isolation & purification ; pharmacology ; Calorimetry ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Escherichia coli ; drug effects ; Gallic Acid ; analogs & derivatives ; isolation & purification ; pharmacology ; Microbial Sensitivity Tests ; Salicylic Acid ; isolation & purification ; pharmacology ; Shigella dysenteriae ; drug effects ; Staphylococcus aureus ; drug effects