AIM: To isolate and culture endothelial progenitor cells (EPCs) in blood vessel by in vitro amplification from bone marrow of rabbits to provide cytology basis for the implantation of autologous EPCs in the repair of blood vessel endothelium. METHODS: The experiment was performed at the Department of Cardiology, Changzheng Hospital, Second Military Medical University of Chinese PLA from March 2005 to February 2006. ①Dil labeled acetylated low density lipoprotein, FITC labeled BS-1 lectin, mouse anti-human CD34 antibody, rabbit anti-human FIK-1 antibody, mouse anti-human CD133 monoclonal antibody were purchased from molecular probes company, vector company, Biolegend company, Biolegeng company and R&D company. ②Totally 8 New Zealand rabbits were selected to extract the bone barrow. Mononuclear cell was isolated from bone marrow by density centrifugation. With the inoculated density of 1?106/cm2, it was put in the M199 medium containing vascular endothelial growth factor and Basic fibroblast growth factor (bFGF) for in vitro cultivation for 7 days. Cell growth and reproductive activity were observed. ③EPCs were identified by Dil labeled acetylated low density lipoprotein and FITC labeled BS-1 lectin. The cells showed red fluorescence were cells phagocytized acetylated low density lipoprotein, while those with green gluorescence were cells bind with BS-1, and the cells double stained showed orange fluorescence. ④Expressions of CD133, CD34 and Flk-1 were detected with immunofluorescent staining and flow cytometry. RESULTS: ①Observation of cell morphous: New isolated mononuclear cells were round. After cultivation for 72 hours, adherent cells showed colony-like growth, and the cells were round or irregular, and the caryocinesis was relatively obvious. Till the 7th day after cultivation, cell colony was connected each other, showing fusiform endothelioid cells. ②Reproductive activity of EPCs in blood vessel: At days 2-4, the reproduction of EPCs was rapid, and then became slow gradually. Growth curve showed typical "S" shape. At days 6 and 7, the EPCs grew rapidly. The absorbance (A) reached 0.58?0.15 and 0.62?0.23, respectively. ③Result of EPCs identification by Dil labeled acetylated low density lipoprotein and FITC labeled BS-1 lectin: In kytoplasm of EPCs, red fluorescent concentration bind with acetylated low density lipoprotein appeared, with the positive rate of over 95%. Combined rate with BS-1 lectin reached 100% nearly. Double staining rate reached over 90%. ④Result of EPCs immunohistochemical method and flow cytometry: The cell-surface marker CD133, FlK-1 and CD34 were positive. CONCLUSION: Cell colony with the feature of EPCs can be isolated and cultured from rabbit bone marrow by in vitro amplification method successfully.

OBJECTIVE To study the drug-resistant diversity of Gram-negative bacilli isolated from inpatients during recent five years.METHODS A total of 1 464 Gram-negative bacilli isolated were detected and retrospectively analyzed from 1999 to 2003.Antimicrobial susceptibility testing was performed using Kirby-Bauer method.RESULTS The resistance of Pseudomonas aeruginosa to piperacillin rised from 17.6% of 1999 to 79.2% of 2003,and that to ciprofloxacin rised from 4.3% of 1999 to 36.0% of 2003.The resistance of Escherichia coli to quinolones was above 50%,while to third-generation cephalosporins was 30-40%;the resistance of E.coli to piperacillin rised from 42.9% of 1999 to 68.9% of 2003,and that to ciprofloxacin rised from 40.0% of 1999 to 73.5% of 2003.The resistance of Acinetobacter to piperacillin rised from 31.2% of 1999 to 67.5% of 2003,and that to ceftriaxone rised from 36.0% of 1999 to 74.1% of 2003.The resistance of Serratia to ceftazidime,ceftriaxone,gentamicin,amikacin and piperacillin rised sharply.Imipenem was the most active antibiotic tested against Gram-negative bacilli.Cefoperazone/sulbactam and piperacillin/tazobactam also showed excellent activity against Gram-negative bacilli.CONCLUSIONS During recent five years,the resistance of the most common Gram-negative bacilli has increased rapidly.How to delay the resistance development of common strains become a global problem.

Objective:To assess the effect of auto-bone marrow mononuclear cells (BMMNCs) transplantation on reendothelialization and neointima formation in vein grafts. Methods: BMMNCs were extracted from the bone marrows of adult rabbit under sterile environment and were labelled with DAPI before transplanted into vein grafts. Twenty adult rabbits were randomly divided into 2 groups: BMMNCs transplantation group(groupⅠ,n=10) and PBS transplantation group (group Ⅱ, n=10). The left external jugular vein of 20 rabbits were harvested and transplanted between ipsolateral common carotid artery(AVG). Three days later, animals in group Ⅰ were transplanted with BMMNCs(6?108 cells)/100 ?l via periotic veins and those in groupⅡwere injected with 100 ?l PBS. Animals were killed 4 weeks later and graft veins were harvested to observe the reendothelialization and the thickness of vein grafts. Results: We found that the transplanted cells survived and were incorporated into the endothelium of vein grafts in groupⅠ.The endothelium integrity of the vein grafts in groupⅠwas significantly better than that of groupⅡ. The intima thickness of vein grafts in groupⅠwas significantly thicker than that of groupⅡ. Conclusion: BMMCs transplantation therapy may improve reendothelialization of the vein graft and inhibits intimal hyperplasi the vein graft.

Objective To analyze the clinical features,curative effect and prognosis of extra-pulmonary infections by mycoplasma pneumoniae(MP).Methods ELISA was used to detect anti-MP IgM and IgG.Case histories of 226 patients,whose nasopharyneal lotione and respiratory secretions were positive of virus and bacteria respectively,was analysed retrospectively.Results One hundred and eighteen of the 226 (52.2%) cases suffered from extra-pulmonary infection.Of these infections 38.98%,33.89%,21.11% and 17.79% were found in digestive, urinary, cardiovascular systems and serous membrane respectively.All of the cases were improved after treatment with macrolides antibiotics.All cases were MP IgM positive,35.5% cases were IgG positive.Spatum MP positive rate was 32.2%.Positive rate of cold-agglutination test was 47.46%.Conclusions MP infection may cause many extra-pulmonary complications.When multi-organ infections can not be explained with bacterial and viral infections,MP infection should be considered.

Adult ; Embolism, Amniotic Fluid ; etiology ; therapy ; Female ; Humans ; Pregnancy

Aim To investigate the role of captopril in insulin resistance of endothelial cells induced by high glucose.Methods 1 .Improvement effect of captopril on insulin resistance in HUVECs was observed.The HUVECs were seeded in a 6-well plate and were ran-domly divided into 5 groups,namely,control group, IR group,IR together with different Cap concentrations (low,medium and high concentration),respectively. 2.Improvement effect of Cap on insulin resistance was mediated by PPARγin HUVECs.HUVECs were ran-domly divided into 6 groups,namely,control group, control +PPARγinhibitor (PI)(1 .0 μmol · L -1 ) group,IR group,IR +PI(1 .0 μmol·L -1 )group,IR +Cap(1 ×1 0 -5 mol·L -1 ) group,and IR +Cap +PI (1 .0 μmol·L -1 )group.All indicators were detected. Results After HUVECs were incubated with media containing 33 mmol·L -1 of glucose for 48 h,the NO levels were significantly decreased while ET-1 levels were significantly elevated,showing a significant differ-ence between IR group and control group (P <0.01 ). The expression levels of PPARγmRNA and its protein were somewhat up-regulated,but there was no signifi-cant difference between IR group and control group (P>0.05).When the HUVECs in IR group were treated with DMEM containing glucose (33 mmol·L -1 )for 48 h and insulin for 30 min,the expression levels of PPARγmRNA and its protein in Cap groups were simi-lar to those in the IR group,and there was no signifi-cant difference between the two groups (P >0.05 );however, the expression levels of phosphorylated PPARγprotein in Cap groups were increased compared with IR group (P <0.05).The levels of NO were sig-nificantly increased whereas the levels of ET-1 were decreased in Cap groups,which had significant differ-ences compared with IR group (P <0.05).Nonethe-less,pre-treating with GW9662,a PPARγinhibitor, the improvement effects of Cap were markedly abol-ished.Conclusions Captopril could improve high glucose-induced insulin resistance of endothelial cells mediated by PPARγ,and the underlying mechanisms are related to the activation of PPARγ,rather than its expression.

The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).

The influence of inner cell mass (ICM) and trophectoderm (TE) score on pregnancy outcomes in frozen-thawed blastocyst transfer cycles was analyzed. A retrospective analysis of 741 cycles of frozen-thawed blastosysts transfer was performed. All cycles were divided into four groups based on the number and morphological score of blastocysts: S-ICM B/TE B group (n=91), the single blastocyst transfer of ICM B and TE B; D-ICM B/TE B group (n=579), double blastocysts transfer of ICM B/TE B; D-ICM B/TE C group (n=35), double blastocysts transfer of ICM B/TE C; and D-ICM C/TE B group (n=36), double blastocysts transfer of TE B/ICM C. The pregnancy outcomes were compared among the four groups. As compared with D-ICM B/TE C group, the clinical pregnancy rate, implantation rate and multiple pregnancy rate were increased in D-ICM B/TE B group (74.96% vs. 57.14%, 57.43% vs. 37.14%, and 48.62% vs. 25%, respectively, P<0.05 for all). Clinical pregnancy rate and implantation rate in D-ICM B/TE B group were also higher than in D-ICM C/TE B group (74.96% vs. 50%, and 57.43% vs. 33.33%, both P<0.05). Multivariable Logistic regression analysis indicated that ICM score was a better predictive parameter for clinical pregnancy (OR=3.05, CI 1.70-5.46, P<0.001), while the trophectoderm score was a better one for early abortion (OR=0.074, CI 0.03-0.19, P<0.001). Clinical pregnancy rate and multiple pregnancy rate in S-ICM B/TE B group were significantly lower than those in D-ICM B/TE B group (46.15% vs. 74.96%, and 2.38% vs. 48.62%, both P<0.05), but there was no significant difference in the implantation rate between the two groups. It was suggested that the higher score of ICM and TE may be indicative of the better pregnancy outcomes. The ICM score is a better predictor of clinical pregnancy than TE, while TE score is a better one in predicting early abortion. Single ICM B/TE B blastocyst transfer in frozen-thawed cycles can also get satisfactory pregnancy outcomes.

· AIM: To inspect and compare the functional vision of an aspheric intraocular lens (Tecnis) with those of conventional monofocal silicone and acrylic intraocular lens and multifocal intraocular lens (Array).· METHODS: The IOLs were tested in the eye model, which was designed to be optically equivalent to the theoretical eye model. The eye model is a combination of a spherical photographic lens with 35mm focal length ( IOL put in a water cell)and a charge coupled device (CCD) camera. The images constructed by the lenses are observed on a monitor of personal computer and the contrasts of the images are analyzed by using commercial image processing software.· RESULTS: The modulation transfer function of the eye model equals the scale produced by the theoretical eye model. The images constructed by changing the diameter of aperture stop and IOL.· CONCLUSION: The proposed eye model is useful for testing functional vision and for inspecting differences of intraocular lens.

Chelating ligand method has been used to synthesize baicalin-metal (Ni2+, Co2+, Cu2+) complexes (BMC). The composition and structure of BMC were characterized by the element analysis, ultraviolet spectrum (UV), infrared spectrum (IR), mass (MS) and thermal gravitational analysis (TGA). MTT was used to analyze the effects of BMC on SMMC-7721 cell proliferation. PI staining method and Annexin-V/FITC double staining method were used to analyze the effects of BMC on the cell cycle and apoptosis of SMMC-7721 cell. Fluorescence quantitative RT-PCR was used to analyze the expression of BMC on Bcl-2 gene and Bax mRNA, flow cytometry was used to analyze BMC on the expression of Bcl-2 protein and Bax protein. The antineoplastic activity and mechanism of action of BMC was explored comprehensively. The results showed that three new kinds of BMC (molar ratio of 2 : 1) were successfully prepared, the complexes molecular formula are: Na2Ni(C21H16O11)2 x 10H2O, Na2Co(C21H16O11)2 x 8H2O and Na2Cu(C21H16O11)2 x 8H2O. According to the results of cell cycle and apoptosis detection, BMC stopped cells at G0/G1 phase to S phase and G2/M phase. Gene and protein detection showed that under the given concentration and time, BMC can downregulate the expression of Bcl-2 gene in SMMC-7721 cells, and significantly decrease the expression of Bcl-2 protein, at the same time, with the increase of expression of Bax gene, the Bax protein's expression increased significantly. Which indicates that BMC restrain cell proliferation and cell apoptosis by stopping cell cycle, reducing the expression of Bcl-2 and increasing that of Bax; The anti-tumor activities of three kinds of complexes were: baicalin-copper (BC-Cu) > baicalin-cobalt (BC-Co) > baicalin-nickel (BC-Ni) > baicalin (BC), showing the dose-response relationship.
Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cobalt ; Copper ; Drug Delivery Systems ; Flavonoids ; administration & dosage ; pharmacology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Metals ; Nickel ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism