BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

OBJECTIVETo study the chemical constituents in bark of Dictamnus dasycarpus.

METHODIsolation and purification were carried out on silica gel column chromatography, prepared thin layer chromatography and sephadex LH - 20, et al. The structures were identified by spectral analysis.

RESULTTwelve compounds were obtained from bark of D. dasycarpus and the structures were determined as dictamnine (I), fraxinellone (II), skimmianine (III), gamma-fagarine (IV ), beta-sitosterol (V), obacunone (VI), limonin disophenol (VII), fraxinellonone (VIII), wogonin (IX), rutevin (X), kihadinin B (XI), dasycarine (XII).

CONCLUSIONCompounds IX and XI were isolated from genus Dictamnus for the first time, and compound VIII was isolated from the species for the first time.

Alkaloids ; chemistry ; isolation & purification ; Dictamnus ; chemistry ; Flavanones ; chemistry ; isolation & purification ; Limonins ; chemistry ; isolation & purification ; Molecular Structure ; Plant Bark ; chemistry ; Plants, Medicinal ; chemistry

No abstract available.
Electrodes*

OBJECTIVETo investigate the effect of Tongxinluo Capsule (TXLC) on platelet aggregation in patients of coronary heart disease (CHD) with aspirin resistance (AR).

METHODSPatients with AR were screened out from 330 CHD patients, who had regularly taken aspirin (100 mg/d) for more than one month, by testing platelet aggregation level after adenosine diphosphate (ADP) and collagen (COL) induction. They were randomly assigned to three groups: the combined treatment group treated by TXLC + aspirin, the TXL group treated by TXLC alone and the AS group treated by aspirin alone, at the dose of TXLC 3 capsules thrice a day, and that of aspirin 100 mg/d. The therapeutic course for all was one month. Patients' platelet aggregation was measured before and after 1-month treatment.

RESULTSEighty-nine patients with AR were screened out from the 330 CHD patients, the occurrence rate being 26.97%. Platelet aggregation was significantly decreased after 1-month treatment in the combined treatment group and the TXL groups (P < 0.05), but changed insignificantly in the AS group, the difference between the former two and the latter group was statistically significant (P < 0.05).

CONCLUSIONTXLC has definite effect in reducing the ADP + COL induced platelet aggregation.

Adenosine Diphosphate ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Aspirin ; pharmacology ; Collagen ; metabolism ; Coronary Disease ; blood ; drug therapy ; Drug Resistance ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Platelet Aggregation ; drug effects

OBJECTIVE:To create an in vitro harvesting method of culturing a large number of adult bone marrow MSCs(BMSCs) DESIGN,TIME AND SETTlNG:The randomized, controlled study was performed at the Shanghai Institute of Digestive Surgery (Key Laboratory of Education Committee of Shanghai City),as well as Department of General Surgery and Organ Transplantation Center,Ruijin Hospital,Medical College.Shanghai Jiao Tong University from September 2005 to April 2006.MATERIALS:Bone marrow samples were collected from normal persons.who did bone marrow examination at the Department of Hematology,Ruijin Hospital,Medical College.Shanghai Jiao Tong University.Donors were volunteers who signed the informed consent.METHODS:Human BMSCs were harvested using Pemoll gradient centrifugation and adherence method.and then incubated in microcarrier cytodex3.Common monolayer polystyrene was incubated as controls.Cell phenotype and proliferative activity were tested utilizing flow cytometry and MTT.MAIN OUTCOME MEASURES:Collection.incubation,morphology of human BMSCs.and prolireration and cell cycle of human BMSCs on the cytodex 3 were measured.RESULlTS:Flow cytometry detection showed that the surface marker of human BMSCs on the cytodex3 was ldentical to that on the common monolayer polystyrene;BMSCs were positive for CD29,CD44 and CD105.but negative for CD14,CD34,CD45,VLA-1 and HLA-DR.MTT detection demonstrated that human BMSCs were in the adaptive phase at days 1-3.and entered logarithmic phase frOm day 3.No significant difference was detected in human BMSCs on the monolayer polystyrene and cytodex3(P＞0.05).On the monolayer polystyrene,human BMSCs entered degenerating stage from day 6,whereas on the cytodex3,human BMSCs were still in the logarithmic growth phase at day 9(P＜0.05).Flow cytometry detection confirmed that the cell cycle of human BMSCs was the same both on the monolayer polystyrene and cytodex3 (P＞0.05). CONCLUSION:Using cytodex3 culture technique,a large amount of human BMSCs can be obtained,and the proliferative activity of these BMSCs is good.

OBJECTIVETo evaluate effect of amygdalin on expression of four biomarkers in the animal model of pulmonary fibrosis induced by bleomycin.

METHODSRats were given one dose (5 mg/kg) of bleomycin in bleomycin-treated groups, amygdalin-treated groups and saline in controls by intratracheal instillation exposed surgically. The amygdalin-treated groups rats were treated with intraperitoneal injection of amygdalin (15 mg x kg(-1) x day(-1)). The rats were sacrificed 7, 14 and 28 days after bleomycin administration. Polarized light microscopy and Image-Pro Plus detected I and III collagen expressed in Paraffin-embedded lung sections stained with Sirius red. Surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS) with weak cationic proteinchip (CM10) detected differentially expressed proteins in the pooled serum samples of all groups.

RESULTSConsistent fibrotic responses were found in all bleomycin and amygdalin-tread groups. On the 7th, 14th and 28th day after bleomycin or saline instillation, four differentially expressed proteins were detected in the pooled serum of all groups rats, consisting of 4 proteins with mass/charge ratio of 3530.7, 7043.5, 8332.6 and 9068.0, respectively. Compared with control groups, protein peaks intensity ratio with mass/charge ratio of 3530.7 on 7, 28 d and 7043.5, 8332.6 and 9068.0 on 7, 14 and 28 d was > 2 in bleomycin-treated groups. Compared with amygdalin-treated groups, protein peaks intensity with mass/charge ratio of 3530.7 at 7, 14, 28 d had no change almost, but protein peaks intensity ratio with mass/charge ratio of 7043.5 at 7 d, 8332.6 on 28 d and 9068.0 on 14 d was > 2 in bleomycin-tread groups. All the four protein peaks intensity had no change almost at other point.

CONCLUSIONAmygdalin may reduce the bleomycin-induced increase of differentially expressed protein peak intensities in rat serum.

Amygdalin ; pharmacology ; Animals ; Biomarkers ; blood ; Bleomycin ; adverse effects ; Blood Proteins ; metabolism ; Male ; Pulmonary Fibrosis ; blood ; chemically induced ; Rats ; Rats, Wistar

[ ABSTRACT] AIM:To investigate the antipyretic effect of patchouli oil on lipopolysaccharide ( LPS)-induced fe-ver in rabbits.METHODS:Male rabbits (n=42) were randomly divided into 7 groups according to their body weight and basal body temperature, including control group, model group, western medical positive group, traditional Chinese medical positive group, and high, middle and low doses (2%, 1%and 0.5%) of patchouli oil groups.Subsequently, except the controls, the rabbits were injected with LPS at a dose of 1 mL/kg (2 mg/L) through marginal ear vein to establish rabbit fever model and the rabbits in control group received the same volume of NS.The rabbits in control group and model group were injected with 0.5%Tween-80 0.5 h late, and the rabbits in the other groups were treated with correspoonding drugs. The effect of patchouli oil on the body temperature was observed, and the levels of interleukin-1β( IL-1β) and tumor nec-rosis factor-α(TNF-α) in the serum, and prostaglandin E2(PGE2) and cyclic adenosine monophosphate (cAMP) in the hypothalamus were measured by radioimmunoassay.RESULTS: The body temperature and the levels of IL-1β, TNF-α, cAMP and PGE2 in model group were significant higher than those in control group.Patchouli oil notably inhibited the body temperature in the febrile rabbits.From 1.5 h to 5.5 h after administration, the body temperatures were increased by (1.06 ±1.55), (1.62 ±1.36), (1.38 ±1.22), (0.98 ±0.98) and (0.48 ±0.95) ℃in high patchouli oil group, re-spectively.From 3.5 to 5.5 h after administration, the body temperatures were elevated by ( 1.47 ±0.73 ) , ( 1.15 ± 0.68) and (0.63 ±0.54) ℃ in middle patchouli oil group, respectively.A tendency of downregulation of the elevated body temperatures was observed at every time point after administration in low patchouli oil group.Patchouli oil significantly decreased the levels of TNF-αin the serum and cAMP content in the hypothalamus, and attenuated the elevated tendency of the IL-1βlevel in the serum and PGE2 level in the hypothalamus.CONCLUSION:Patchouli oil evidently has antipyretic effect on LPS-induced fever in the rabbits.The antipyretic mechanism might be related to the inhibition of TNF-αlevel in serum and cAMP content in the hypothalamus.

Objective To study the effects of fluorine and aluminum on index of hematologic tests of rats. Methods According to body mass,56 Wistar rats of 130-200 g were randomly divided into control,low-fluorine (F),middle-F,high-F,low-F + aluminum(Al),middle-F + Al,high-F + Al group,8 rats in each group were given a series of doses of fluoride and aluminum,which were (0 + 0),(100 + 0),(200 + 0),(300 + 0),(100 + 10),(200 + 10),(300 + 10)mg/L After 90-day intragastrie administration,blood samples were collected on eyes of rats to undergo blood routine test,including red blood cell (RBC),lymphocyte (LYM),platelet (PLT),hemoglobin (HGB),white blood cell (WBC),hematocrit (HCT),mean corpuscular hemoglobin (MCH),mean corpuscular-hemoglobin concentration(MCHC),mean corpuscular volume(MCV),and at the same time some blood biochemistry indicators related to functio ns of liver and kidney were determined such as aspartic acid aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),Crea(Cr) and Urea. Organ coefficient of liver and kidney were calculated. Results The difference of RBC,HCT,MCV among all groups of rats was statistically significant(F = 3.202,3.316,2.915,P < 0.05). The RBC,HCT of the low-F group[(7.59± 2.40)×10~(12)/L,0.51±0.11],the middle-F group[(8.60±1.16)×10~(12)/L,0.55±0.05],the high-F group[(9.23± 0.60)×10~(12)/L,0.54±0.03],the low-F + Al group[(9.25±0.79)×10~(12)/L,0.53±0.04],the middle-F + Al group[(7.98±2.14)×10~(12)/L,0.49±0.08]and the high-F + Al group[(7.61±3.17)×10~(12)/L,0.49±0.16]were significantly higher than that in the control group[(4.46±3.10)×10~(12)/L,0.31±0.16,P< 0.05 or < 0.01)]. The MCV of the middle-F group[(64.06±6.51)fl],high-F group[(58.67±1.13)fl],low-F + Al group[(57.78± 1.57)fl]and the middle-F + Al group[(63.04±10.64)fl]were significantly higher than the control group[(78.54± 15.57)fl,P < 0.05 or < 0.01]. The difference of AST and Urea among all the groups of mrs serum was statistically significant(F= 2.847,5.549,P < 0.05 or < 0.01). The serum AST of low-F group[(399.00±54.99)U/L],the middle-Fgroup[(465.60±76.99)U/L],the high-F group[(465.80±75.41)U/L],the low-F + Al group[(346.00±69.26) U/L],the middle-F + Al group[(437.40±68.31)U/L]and the high-F + Al group[(403.00±30.61)U/L]were all significantly higher than that in the control group[(336.67±94.34)U/L,P < 0.05],and the high-F group significantly higher than the high-F + Al group(P < 0.05). The serum Urea of the middle-F group[(7.70±0.52)mmol/L],the high-F group[(8.44±1.30)mmol/L],the low-F + Al group[(7.83±0.62)mmol/L],the middle-F + Al group [(7.73±0.47)mmol/L],and the high fluoride + aluminum group[(7.70±0.21)mmol/L]were all significantly higher than that in the control group[(6.55±0.50)mmol/L,P< 0.05 or < 0.01],and the low-F group was significantly lower than the low-F + Al group(P < 0.01),however the high-F group was significantly higher than that in the high-F + Al group(P< 0.05). The liver organ coefficient of the low-F group(2.94±0.36) was higher than the low-F + Al group (2.60±0.15,P < 0.05). Conclusions Fluorine and combination of aluminum and fluorine have toxicity on rats to a certain extent,including the proliferation of crythrocytes of rat,while the cell size gets smaller and the cell quality is deteriorated,meanwhile functions of liver and kidney are impaired. Aluminum shows different joint action in different concentrations of fluorine.

OBJECTIVETo evaluate the safety of iodine-125 seed implantation in the liver.

METHODSTwenty New Zealand rabbits were divided into control and treatment groups and in the latter, iodine-125 seeds of 37 MBq were implanted into the liver under CT guidance whereas nonradioactive seeds were implanted in the control rabbits. Four weeks after implantation, white blood cell count, liver functions, and renal functions were measured or evaluated for comparison with those before implantation. The rabbits were then anesthetized to collect the liver tissue for pathological examination with HE staining and cell apoptosis assay.

RESULTSObvious hepatic tissue necrosis was observed around the radioactive seeds in the treatment group. At a 5 mm distance to the seeds, a distinct boundary occurred between the necrotic hepatic cells and normal cells. The control rabbits, however, had normal liver structure around the seeds implanted. In situ cell apoptosis examination showed a distinct band of apoptotic cells in the liver tissue of rabbits in the treatment group, which was not found in the control group. Two weeks after iodine-125 irradiation, alanine aminotransferase significantly increased in the treatment group (t=6.285, P<0.001), but recovered two weeks later (t=2.002, P=0.06). No significant alterations occurred in aspartate aminotransferase, blood urea nitrogen, serum creatinine, hemoglobin, serum total bilirubin, white blood cell count, or platelet count after the seed implantation.

CONCLUSIONIodine-125 seed implantation in the liver results in conformal irradiation dose distribution without obvious effects on the vital organs, demonstrating iodine-125 seed implantation as a safe and minimally invasive technique for hepatic cancer treatment.

Alanine Transaminase ; blood ; Animals ; Apoptosis ; radiation effects ; Dose-Response Relationship, Radiation ; In Situ Nick-End Labeling ; Iodine Radioisotopes ; adverse effects ; Liver ; pathology ; physiopathology ; radiation effects ; Male ; Rabbits ; Radiation Injuries, Experimental ; blood ; etiology ; pathology ; Random Allocation ; Time Factors

AIMTo study the gene expression profiles between the CCl4 injured liver and normal liver in mice, and to screen the differentially expressed genes that relate to liver injury by CCl4 on a large scale using cDNA microarrays.

METHODSMale Kunming strain mice were divided into two groups: one was control group and another was CCl4 injured liver group that was given 0.1% CCI4 oil solution ip at dose of 10 mL x kg(-1) every three days, totally for ten times. Then mRNA in livers of the two groups of mice was extracted, separately, and reversely transcribed to cDNA with the incorporation of different fluorescent-labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarrays. The fluorescent signal values were acquired by scanner and analyzed with statistical software.

RESULTSAmong the 14 100 target genes, 379 genes were differentially expressed, in which 163 genes were up-regulated and the other 216 genes were down-regulated. They are closely related to a range of biological functions.

CONCLUSIONUsing the cDNA microarray and experimental animal modeling technique, the differentially expressed genes of CCl4 injured liver in mice on a large scale could be studied. It is useful for further investigation of the injury mechanism of CCl4.

Alanine Transaminase ; blood ; Animals ; Apoptosis Regulatory Proteins ; metabolism ; Aspartate Aminotransferases ; blood ; Carbon Tetrachloride Poisoning ; Chemical and Drug Induced Liver Injury ; etiology ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Gene Expression Profiling ; Liver ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 12 ; metabolism ; Mice ; Nuclear Proteins ; metabolism ; Oligonucleotide Array Sequence Analysis ; methods ; Proteins ; metabolism ; Random Allocation