Objective To investigate the effects of post-operative analgesia with oxycodone or morphine for patients undergoing colon cancer radical surgery on platelet activation and cellular immunity.Methods Forty colon cancer patients scheduled for radical surgery, 23 males and 17 females, ASA physical status Ⅰ or Ⅱ, were randomly divided into 2 groups (n=20 each): oxycodone group (group O) and morphine group (group M).Patient-controlled intravenous analgesia (PCIA) was used for post-operative analgesia.PCIA solution contained oxycodone 1 mg/kg and tropisetron 6 mg in 100 ml normal saline in group O or morphine 1 mg/kg and tropisetron 6 mg in 100 ml normal saline in group M.Blood samples were obtained from the patients at 5 min before anesthesia induction (T0), 4 h after surgery (T1), 24 h after surgery (T2) and 48 h after surgery (T3).The levels of glycoprotein (GP)Ⅱb/Ⅲa, P-selection (CD62P), natural killer (NK) cells, NKT cells, and natural Treg (nTreg) cells were detected.The platelet aggregation rate (PAR) was determined.Results Compared with T0, the levers of GPⅡb/Ⅲa, CD62P, PAR and nTreg cells were significantly higher at T1 in group O and at T1, T2 in group M (P<0.05).Compared with T0, the levels of NK and NKT cells were decreased significantly at T1 in group O and at T1-T3 in group M (P<0.05).The levels of GPⅡb/Ⅲa, CD62P, PAR and nTreg cells at T2 and T3 in group O were decreased significantly as compared with group M (P<0.05).The levels of NK cells, NKT cells at T2 and T3 in group O were significantly higher than those in group M.Conclusion Post-operative analgesia with oxycodone for patients undergoing colon cancer radical surgery exhibits a more significant effect of decreasing platelets activity and presents a less disturbance on cellular immunity as compared with morphine.

To establish a method for detecting microdialysis recovery of tetramethylpyrazine (TMP) and ferulic acid (FA) and investigating the influencing factors, providing the basis for further in vivo microdialysis experiments. The concentration of FA and TMP in dialysates were determined by high pressure liquid chromatography ( HPLC) and probe recovery were calculated respectively. The influence of the flow rates, medium concentration, temperature and in vivo probe stability on the recovery of FA and TMP were investigated by using concentration difference method (incremental method and decrement method). The recovery obtained by incremental method were similar to by decrement method. The in vitro recovery rate of FA and TMP decreased with the increase of 1-2.5 μL min(-1), and increased obviously with the temperature of 25-42 degrees C under the same conditions. The concentration of FA and TMP had no obvious effect on the probe recovery under the same flow rate. In addition, the recovery of TMP and FA remained stable and showed similar trends under the condition of four concentration cycles, indicating that the intra day reproducibility of the concentration difference method was good. The recovery of brain microdialysis probes in vivo 8 h maintained a relatively stable, but certain differences existed between different brain microdialysis probes, demonstrating that each probe was required for recovery correction in vivo experiment. Microdialysis sampling can be used for the local brain pharmacokinetic study of FA and TMP, and retrodialysis method can be used in probe recovery of FA and TMP in vivo.
Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Coumaric Acids ; analysis ; isolation & purification ; pharmacokinetics ; Drugs, Chinese Herbal ; Humans ; Microdialysis ; methods ; Pyrazines ; analysis ; isolation & purification ; pharmacokinetics ; Rats

OBJECTIVETo establish a new method for simultaneous determination of shanzhiside methyl ester, chlorogenic acid, 8-O-acetyl shanzhiside methylester, forsythiaside B, rutin, acteoside and galuteolin in Lamiophlomis rotata.

METHODSeparation was performed on a Welchrom-C18 chromatographic column with acetonitrile-0.1% orthophosphoric acid as mobile phasewith gradient elution. The flow rate was 1.0 mL x min(-1). The column temperature was 30 degrees C, and the detection wavelength was set at 238 nm, 330 nm and 350 nm.

RESULTThe seven compounds were well separated with good linear correlations. The mean recoveries of seven compounds were 96.47%-102.2% (RSD 0.70%-2.2%).

CONCLUSIONThere were good correlations among the seven compounds in the samples of aerial parts. The mean sum of shanzhiside methyl ester and 8-O-acetyl shanzhiside methylester in samples of aerial parts is 1.44%. The aerial parts have more kinds of composition and with higher content than that of underground parts in L. rotata, which was reasonable for the resonable use of the aerial part as medicinal part. The method was simple, repeatable and stable, which could be used for identification and quality evaluation of L. rotata.

Chlorogenic Acid ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Glycosides ; chemistry ; Lamiaceae ; chemistry ; Methyl Ethers ; chemistry ; Phenols ; chemistry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Pyrans ; chemistry ; Rutin ; chemistry

Objective To investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.Methods 0.3 ml fat tissue,derived and refined from clinical liposuction patients.was mixed with different concentrations of SVFs as 5 × 105/ml in Group A,or 1 × 106/ml in Group B,or 2 × 106/ml in Group C,or completely medium in control group D.Then the mixture was injected randomly under the back skin of 6 nude mice.The transplanted fat tissue in four groups was harvested at 3 months after implantation.Wet weight of fat grafts was measured for macroscopic aspects.After HE staining,blood vessel density,viable adipocytes and fibrous proliferation were counted respectively for histological evaluation.Results The wet weight of fat grafts in group B(81.670 ± 7.528) mg was significantly higher than that in group A、C、D [ (60.000 ±6.325 ) mmg、(68.330 ± 7.528 ) mg、(48.330 ± 7.528 ) mg,respectively,P ＜ 0.05 ) ],but the difference between group A and group C was not statistically significant(P ＞ 0.05).The grafts in group A 、B and C had significantly higher blood vessel density than those in the control group D,whereas blood vessel density was the highest in group B( P ＜ 0.05 )and there was no significant difference between group A and C (P ＞0.05).Compared with group A,C and D,histological analysis revealed that the fat grafts in group B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis (P < 0.05 ).However,fibrosis counts were significant lower in group A,B and C than those in group D( P <0.05),and there was no significant difference between group A and C(P ＞0.05).Conclusions The human isolated SVFs has the advantages to improve the survival rate of fat transplantation,and the magnitude of 1 × 106/ml is more practical and safe,indicating a wide clinical application in the future.

Objective To investigate the effect of different concentrations of human adipose stromal vascular fraction cells (SVFs) on the survival rate of fat transplantation.Methods 0.3 ml fat tissue,derived and refined from clinical liposuction patients.was mixed with different concentrations of SVFs as 5 × 105/ml in Group A,or 1 × 106/ml in Group B,or 2 × 106/ml in Group C,or completely medium in control group D.Then the mixture was injected randomly under the back skin of 6 nude mice.The transplanted fat tissue in four groups was harvested at 3 months after implantation.Wet weight of fat grafts was measured for macroscopic aspects.After HE staining,blood vessel density,viable adipocytes and fibrous proliferation were counted respectively for histological evaluation.Results The wet weight of fat grafts in group B(81.670 ± 7.528) mg was significantly higher than that in group A、C、D [ (60.000 ±6.325 ) mmg、(68.330 ± 7.528 ) mg、(48.330 ± 7.528 ) mg,respectively,P ＜ 0.05 ) ],but the difference between group A and group C was not statistically significant(P ＞ 0.05).The grafts in group A 、B and C had significantly higher blood vessel density than those in the control group D,whereas blood vessel density was the highest in group B( P ＜ 0.05 )and there was no significant difference between group A and C (P ＞0.05).Compared with group A,C and D,histological analysis revealed that the fat grafts in group B was consisted predominantly of adipose tissue with less fat necrosis and fibrosis (P < 0.05 ).However,fibrosis counts were significant lower in group A,B and C than those in group D( P <0.05),and there was no significant difference between group A and C(P ＞0.05).Conclusions The human isolated SVFs has the advantages to improve the survival rate of fat transplantation,and the magnitude of 1 × 106/ml is more practical and safe,indicating a wide clinical application in the future.

OBJECTIVETo explore the possibility of building tissue-engineered adipose tissue and looking for a new approach for the repair of soft tissue defects.

METHODSThe cells using enzymatic digestion from human liposuction part of the lipid extract were used as adipose tissue-derived cells and labeled with DiI fluorescent marker, the induced group using I collagen scaffold material as a carrier, the induced cell were planted into left back subcutaneously in nude mice at 1 x 10(7)/ml cell density, in the uninduced group cells were not induced by any, in the same cell density and type I collagen scaffold composite inoculated in nude right mouse back skin, the blank control group I collagen scaffold gaps in nude mice inoculated subcutaneously center of the neck, each of the six mice; Remove implants after 12 weeks and judge the adipogenic capacity through general and fluorescence microscopy, wet - determination, histological detection and oil red O staining qualitative.

RESULTSThe primary source of fat cultured stem cells, similar to the fibroblast morphology, and has a strong proliferative capacity. In the role of adipose differentiation medium, it can be the mature fat cells in which cytoplasmic lipid droplets gather, oil red O staining was positive. In the induced group, newborn tissue were found in the experimental groups of nude mice and its average weight is about 0.020 g. Conventional pathological slices and oil red O staining confirmed it is mature adipose tissue, the fluorescence staining positive confirm them are exogenous. Uninduced group newborn tissue are found in the experimental groups of nude mice and its average weight is about 0.014 g. Conventional pathological slices and oil red O staining confirmed it include some mature adipose tissue, the fluorescence staining positive confirm them are exogenous. Two groups of the new wet weight with have statistical significance (P < 0.01); gaps in the control group no new organization formed.

CONCLUSIONSThe cells using enzymatic digestion from human liposuction part of the lipid extract are adipose tissue-derived cells. The cells can be as seed cells and with solid scaffold of collagen type I it can become fat tissue in vivo successfully.

Adipose Tissue ; cytology ; metabolism ; Animals ; Cell Culture Techniques ; Collagen Type I ; biosynthesis ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Stem Cells ; cytology ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo study the cellular compatibility of type I collagen scaffold and human adipose-derived stem cells (ADSC(S)) in order to explore appropriate scaffold materials for adipose tissue engineering.

METHODSThe morphology and function of the ADSC(S) were observed by inverted phase contrast microscope, scanning electron microscope and XTT assay when cocultured type I collagen scaffold with ADSC(S) in vitro. Cells adhesive rates were also calculated.

RESULTADSC(S) were able to attach, grow and proliferate well on the scaffolds.

CONCLUSIONcollagen I scaffold exhibits excellent cellular compatibility and can be used as a vehicle for adipose tissue engineering.

Adipocytes ; cytology ; Adult Stem Cells ; cytology ; Biocompatible Materials ; chemistry ; Cells, Cultured ; Coculture Techniques ; Collagen Type I ; chemistry ; Humans ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo investigate the effects of chronic lead exposure on expression of autophagy-associated proteins in rat hippocampus.

METHODSSD rats were randomly divided into three groups: control group was given distilled water, lead-exposed groups were given 0.5 g/L (low-dose) or 2.0 g/L(high-dose) lead acetate solution in drinking water. The rat pups started to drink the lead content water until 60 d maturity. The lead contents in blood and brain samples were analyzed by graphite furnace atomic absorption spectrophotometry. The expressions of Beclin 1, LC3, LAMP2 and cathepsin B proteins were detected by Western blot and immunohistochemistry.

RESULTSCompared with control group, the contents of lead were significantly higher in blood and hippocampus samples in chronic lead-exposed rats (P<0.01). Western blot showed that the expression of Beclin 1 and LC3-II/LC3-I increased significantly in high dose lead-exposed group compared with control group (P<0.05 or P<0.001). The confocal laser immunostaining results demonstrated that increased immunofluorescence staining of cathepsin B in hippocampal neurons compared with control animals.

CONCLUSIONThe disturbance of autophagy-lysosome signaling molecules might be partially contribute to neurotoxicity of chronic lead exposure.

Animals ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; drug effects ; physiology ; Beclin-1 ; Cathepsin B ; metabolism ; Chronic Disease ; Disease Models, Animal ; Female ; Hippocampus ; drug effects ; metabolism ; pathology ; Lead Poisoning ; metabolism ; pathology ; Lysosomal-Associated Membrane Protein 2 ; metabolism ; Male ; Microtubule-Associated Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects

OBJECTIVETo explore the migration of intravenously injected adipose tissue-derived stem cells (ADSCs) in SD rats with soft tissue wound.

METHODS1.8 cm x 1.8 cm full thickness skin defect and 0.5 cm in depth soft tissue defects were made on the back of 6 SD rats. ADSCs were isolated and cultured in vitro for 3 passages. 2.4 x 10(6) ADSCs were labeled with DiI and transplanted into the SD rats through tail vein. Normal skin and wound tissue samples were collected for fluorescent distribution observation 24, 48 days after injection, respectively.

RESULTSCompared to normal skin, more fluorescent positive cells were detected in the margin and deep layer of the wound 24 days after operation. But it is accumulated within dermis and adenoid 48 days after operation.

CONCLUSIONWound can probably induce the migration and accumulation of intravenously transplanted ADSCs.

Adipocytes ; cytology ; Animals ; Cell Movement ; Male ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; Stem Cells ; cytology ; Wound Healing ; Wounds and Injuries

In this study, the effects of apollon antisense oligodeoxynucleotide (ASODN) on the proliferation and apoptosis of human Lovo cells in vitro were investigated. Apollon ASODN was incubated with human colorectal Lovo cells for 48 h, the proliferation inhibition and the clone forming rates were detected by WST method and clone formation assay, respectively. The expression of apollon mRNA was analyzed by real time fluorescent quantitative reverse transcription polymerase chain reaction. The percentage of apoptotic cells and cell cycle distribution were determined by flow cytometry. The morphology of apoptotic cells was examined by fluorescence microscope. Lovo cells incubated with apollon ASODN combined with 5-fluorouracil (5-FU), cisplatin (DDP) or epirubicin (EPI) of different concentrations, cell proliferation inhibition rates were detected with WST method and IC50 was calculated. It was found that ASODN targeting apollon gene could all suppress the growth of Lovo cells and induce apoptosis of these cells significantly (P < 0.05). After Lovo cells treated with apollon ASODN for 48 hours, the expression of the apollon mRNA level was suppressed significantly. And a marked concentration-dependent decline of cell proliferation and clone forming, increasing of cell apoptosis levels were observed. The percentage of G0/G1 phage cells was abated and that of S phage cells was increased and the Lovo cells arrested at S phage of the cell cycle detected with flow cytometry. Many Lovo cells stained with Hoechst 33258 exhibited apoptotic morphology such as cell shrinkage, nuclear condensation and nuclear fragmentation. Cell proliferation inhibition was detected and their chemo-therapeutic effects of 5-FU, DDP and EPI on Lovo cells combined with apollon ASODN (0.08 micromol x L(-1)) were enhanced independently compared with single 5-FU, DDP and EPI groups, and the sensitivity enhanced about 2.58, 4.47, and 5.33 times respectively. It can be concluded that ASODN targeting apollon can suppress the expression of apollon mRNA, and inhibit the proliferation, induce apoptosis, arrest cell cycle at S phase of colorectal cancer Lovo cells in vitro and enhance the chemo-sensitivity to 5-FU, DDP and EPI.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Colonic Neoplasms ; metabolism ; pathology ; Epirubicin ; pharmacology ; Fluorouracil ; pharmacology ; Gene Knockdown Techniques ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Inhibitory Concentration 50 ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; metabolism ; Sensitivity and Specificity ; Transfection