BACKGROUND: Longterm therapeutic effects of routine drug treatment, intervention or vascular bypass transplantation on lower extremity arterial occlusion are not ideal. During recent years, angiogenesis of stem cells possibly becones a new method to repair or rebuild an effective collateral circulation at infarct regions.OBJECTIVE: To verify the effect of autologous bone marrow mesenchymal stem cell transplantation on promoting angiogenesis in rabbit ischemic hind limbs.DESIGN, TIME AND SETTING: A randomized controlled animal study was performed in Jiangsu Institute of Schistosomiasis Control between August 2005 and November 2006. MATERIALS: Eight rabbits were used to prepare ischemic models of hind limbs, and then they were randomly divided into experimental group (n=4) and control group (n=4).METHODS: Bone marrow mesenchymal stem cells were isolated and cultured from New Zealand rabbits in the experimental group, and they were then marked 5-bromo-2-deoxyuridine (Brdu). Suspension of bone marrow mesenchymal stem cells was injected into the ischemic hind limbs in the experimental group, while the same volume of saline was injected into the controls. MAIN OUTCOME MEASURES: After two weeks, two-dimensional and color Doppler ultrasound detection was used on rabbit femoral artery to measure inner diameter of blood vessel, peak velocity and acceleration time of blood flow before and after transplantation. Muscle tissues were obtained from ischemic regions to observe distribution of transplant cells and state of angiogenesis using immunofluorescence staining and hematoxylin-eosin (HE) staining. RESULTS: Two weeks after transplantation, the inner diameter of femoral artery and the peak velocity of blood flow in the experimental group were higher than those in the control group (P < 0.01), but the acceleration time of blood flow was shorter than that in the control group (P < 0.01). Lmmunofluorescence staining showed that anti-Brdu-staining positive cells were found out in transplant part in the experimental group; while HE staining indicated that vessel density of ischemic region in the experimental group was higher than that in the control group. CONCLUSION: Bone marrow mesenchymal stem cells can promote angiogenesis, while autologous bone marrow mesenchymai stem cell transplantation will become a simple and effective method to treat lower limb ischemia.

BACKGROUND:Several studies have demonstrated that stem cells can differentiate into vascular endothelial cells, and then further differentiate and form blood capillary. Based on this principle, autologous bone marrow mesenchyrnal stem cell (BMSC) transplantation promotes angiogenesis to treat ischemia in lower limb.OBJECTIVE: To evaluate the angiogenesis in rabbit ischemic limbs following autologous BMSC transplantation using high-frequency two-dimensional ultrasound detection in conjunction with Doppler color-flow imaging examination. DESIGN, TIME AND SEI-FING: A randomized, controlled, animal experiment was performed in the Third People's Hospital of Wuxi between March 2007 and April 2008.MATERIALS: Twenty-four New Zealand rabbits were randomized to a control group and a cell transplantation group, with 12 rabbits in each.METHODS: Ischemia in lower limbs was induced in all rabbits. One week following ischemia induction, the cell transplantation group was injected with 0.5 mL cell suspension, comprising 2×106 BrdU-labeled autologous BMSCs cultured in vitro, through multiple sites in the region of gastrocnemius muscle. Simultaneously, the control group received the same amount of physical saline in the same region.MAIN OUTCOME MEASURES: The initial segment of rabbit femoral artery and superficial femoral artery was subjected to high-frequency two-dimensional ultrasound and Doppler color-flow imaging examinations to measure femoral artery vascular internal diameter, blood flow peak velocity, and blood flow acceleration time. Ischemic muscular tissue was taken for immunohistochemical staining to detect transplanted cell distribution and for pathological examination of angiogenesis. RESULTS: Two weeks following autologous BMSC transplantation, high-frequency ultrasound results revealed that femoral artery internal diameter and blood flow peak velocity were greater, but blood flow acceleration time was shorter, in the cell transplantation group than in the control group (P<0.01). Immunohistochemical staining results demonstrated the presence of BrdU-positive cells. Pathological sections displayed that vascular density was significantly higher in the cell transplantation group than ih the control group. CONCLUSION: Autologous BMSC transplantation is a promising, simple, and effective method of treating ischemia in lower limbs owing to its promotion of angiogenesis. Meanwhile, high-frequency ultrasound detection of femoral artery is an effective, practical method to evaluate the clinical outcomes of autologous BMSC transplantation.

BACKGROUND: There have been no effective means for clinical treatment of large regions of bone defects.Nano-hydroxyapatite/collagen(nHAC)composite would provide a new pathway for repair of bone defects owing to its similar structure to natural skeleton and better biocompatibility.OBJECTIVE: To investigate the role of nHAC composite co-cultured with bone marrow-derived mesenchymal stem cells(BMSCs)in repair of bone defects.METHODS: Following isolation and culture,human BMSCs were co-cultured with nHAC composite.Gross observation,histological analysis,and electron microscope observation were performed to analyze osteogenesis for repair of bone defects in the clinic.RESULTS AND CONCLUSION: Human nHAC could greatly proliferate in vitro.X-ray photography revealed that bone defects well healed after implantation of nHAC/BMSCs composite.These findings indicate that BMSCs exhibit osteogenic potential and nHAC is a satisfactory scaffold material for construction of tissue-engineered bone.

Background and purpose:Endostatin can effectively inhibit angiogenesis,it could slow down tumor growth in heterogenic transplantation model by given ES systematically.This study was to evaluate the inhibition of endostatin on tumor growth and metastasis of lung cancer of the thoracic cavity implant model in nude mouse.Methods:H460 cells were inoculated subcutaneously into the thoracic cavity of nude mouse,and the mice were randomized into ES group(ESG)and control group(CG).At the fi rst day,the ESG was given 20 mg/kg of endostatin s.c.qd,the CG was treated daily s.c.with equal volumes of PBS for 20 days.The weight of the animal and the transplantation tumor,mice life span,MMP2 of blood were observed.Results:The rate of tumor formation was 100%,and the thoracic cavity implant models gradually had metastasis.The transplantation tumor weights were(1.54?0.14)in ESG group that was less than that(0.9?0.3)in CG group(t=-3.163,P=0.005);the comparison of the change value of weight on day 1 and day 21 between CG(1.9?2.8)and ESG(-0.8?2.8)was statistically different(t=-2.156,P=0.045).The average survival time of the ESG was longer than that of the CG.Conclusion:The results demonstrate that endostatin could inhibit the growth and metastasis of H460 cell in athymic mouse,and one of the important mechanisms involves was inhibition of the MMPs expression.The thoracic cavity implant model was easier to obtain metastasis,and provided a reliable research tool for the study of endostatin inhibition on tumor growth and metastasis of lung cancer.

BACKGROUND: Bone bone bone marrow derived mesenchymal stem cells (MSCs) has the capacities of self-renewal and multi-directional differentiation, which can differentiate into osteoblasts after induction. Combined with basal hydroxyapatite materials, MSCs can repair the bone defect with satisfactory shape and function.OBJECTIVE: To observe the bone formation performance of segmental bone defect repaired by autologous transplantation after combination of MSCs and basal hydroxyapatite materials.DESIGN: Open experiment.SETTING: Department of Cells, Third People' s Hospital of Wuxi.MATERIALS: The experiment was conducted in the Department of Cells,Third People's Hospital of Wuxi between May 2004 and March 2005.Eighteen New Zealand rabbits of clean grade, aged 6 months were selected and randomly divided into cytoskeleton group, simple scaffold group and blank control group with 6 rabbits in each group. Nano-collagen basal hydroxyapatite bone were provided by the Department of Material, Tsinghua University, which is characteristic of natural-bone microstructure. Composition: about 57% were hydroxyapatite,and 30% were collagen and 13%were polylactic acid.METHODS: ①bone bone marrow derived MSCs of rabbits were isolated and cultured. Well-grown cells of the second generation were inspected of the expression of cell surface antigen (CSA) with EPICS-ALTRA flow cytometry. ②Scaffold materials were cut into blocks with the size of 6 mm ×6 mm×4 mm, sterilized by 60Co irradiance and infiltrated with DMEM-LG before using. Cells of the 2nd and 3rd generations were collected, cultured,the concentration of which was regulated to 109 L-1. Cells were co-cultured with nano-collagen basal hydroxyapatite in vitro. ③Model of segmental defects in radial of rabbits were established in all groups. The prepared compounds in the cytoskeleton compound group were embedded in the bone defect according to autograft principle, and blank scaffold materials were embedded in the bone defects of simple scaffold group. Nothing was transplanted in the blank control group. Rabbits in all groups received no internal or external fixation, whose soft tissues and skins were closely sutured.④The surface structure of materials and cell adherence in all groups were observed under scanning electron microscope (SEM).⑤Bone formation was studied by general observation, histological analysis and X-rays at 4, 8 and 16 weeks after operation respectively.MAIN OUTCOME MEASURES: ①The isolated culture and identification of MSCs.②Observation of all groups under SEM.③Results of radiological inspection of all groups. ④General morphous of cells in each group after operation.⑤Results of histological inspection of all groups after operation.RESULTS: A total of 18 enrolled New Zealand rabbits were involved in the analysis of results.①There were adherent cells at 4 hours after primary culture, which were like cloned at 48 hours after static culture. Cells were in irregular scale-shape with large cell body and the nucleus was in middle. Inspection with flow cytometer showed that CD29,CD44, CD-105 were positive, and the ratios of three positive cells were 97.4% ,98.1% and 86.2% respectively. ②The surface of simple scaffold group was irregular with many ventages. There were cells adherent to the surface of materials as well as the inside of ventage in the cytoskeleton compound group. ③The absorbance of X-rays at 4, 8 and 16 weeks after operation were higher in the cytoskeleton compound group than blank control group and simple scaffold group, moreover, bone union could be seen at the 16th week in the cytoskeleton compound group. ④The implants was embedded in the osteotylus at 16 weeks after operation in the cytoskeleton compound group,while parts of the middle segment of implants in the simple scaffold group was not covered by bone tissues. No porosis was found in the blank control group.⑤The ossifying capacity at each time point after operation was better in the cytoskeleton compound group than simple scaffold group, moreover,there was bone trabecula formed at the 16th week, and the osteoblasts were arranged in order. The bone tissues in the simple scaffold group mainly concentrated on the surface of materials, which sucked little. There were some bony tissues formed in the proximal end of defects in the blank control group, while none was seen in the middle parts.CONCLUSION: Rabbit MSCs can greatly proliferate in vitro, which has strong osteogenesis by being co-cultured with nano-collagen basal hydroxyapatite. It is a good kind of seed cells in tissue engineering.

Objective To evaluate the value of procalcitonin(PCT)in different periods for diagno-sis of early-onset of neonatal bacterial infection.Methods One hundred and ninety-five newborns with intra-uterine infection risk factors were divided into two groups:infection group(24cases)and non-infection group(171cases).The levels of PCT,C-reactive protein(CRP)and WBC were measured in 2hours,6to 12hours,12to 36hours and more than 48hours after birth.The sensitivity and specificity of PCT in different periods in the diagnosis of early-onset infection were analyzed.Results There were no significant differ-ences in the positive rate of PCT,CRP and WBC in infection group in 2hours after birth(P﹥0.05).The sensitivity and specificity for diagnosis of early-onset infection of PCT were 91.7% and 86.5% at 6to 12hours after birth,which were higher than those of CRP and WBC.After birth in 12to 36hours was the physiologic peak of PCT,so it couldn′t have higher sensitivity and specificity.According to threshold of 0.5ng/ml,2ng/ml,and 10ng/ml for PCT,the sensitivity was 100%,91.7% and 100% respectively,and the specificity was 5.8%,53.8%and 95.9%respectively.Conclusion PCT in 6to12hours after birth,ac-cording to threshold of 2ng/ml,can reach higher sensitivity and specificity for diagnosis of early-onset neo-natal bacterial infection.

ObjectiveTo investigate the incidence and pathogen distribution of neonatal early-onset sepsis (EOS) following exposure to antenatal antibiotics.MethodsOne hundred and eighty-four neonates who were admitted to Maternal and Child Care Hospital of Xiamen and identified as having EOS from January 2010 to December 2015 were enrolled. The clinical data were retrospectively analyzed. According to antenatal antibiotic exposure time, the infants were divided into the antibiotics group (≥4 hours) and the control group (<4 hours). Women in late pregnancy (35-37 weeks of gestation) underwent group BStreptococcus (GBS) screening using standard bacterial culture beginning from Janaury 2014 as screening group. Intrapartum antibiotic prophylaxis was given if the GBS culture was positive. Infants delivered before January 2014 were included in the no-screening group. Pathogen distribution and the difference in drug resistance between the two groups were compared by a two-independent samplest-test andChi-square test.ResultsIn the antibiotics group, the percentages of birth weight lower than 2 500 g, preterm infants, asphyxia, and positive rates of GBS and blood culture were 24.3%(17/70), 14.3% (10/70), 2.9% (2/70), 7.1% (5/70) and 70.0% (49/70), respectively, and were significantly lower than those in the control group [39.5%(45/114), 28.1% (32/114), 14.9% (17/114), 19.3%(22/114) and 88.6% (101/114), respectively] (χ2=4.478, 4.678, 6.807, 5.118 and 9.957, allP<0.05). There was no difference in the positive rate of coagulase-negativeStaphylococci andE. coli culture, or in the incidence of purulent meningitis, septic shock, disseminated intravascular coagulation, hospital stay and fatality rate between the antibiotics group and control group (allP>0.05). Compared with the no-screening group, the positive rate of GBS decreased [7.6% (5/66) vs 18.6% (22/118)] and the positive rate of fungal infection increased [7.6%(5/66) vs 1.7% (2/118)] in the screening group (χ2=4.141,P=0.042;χ2=4.000,P=0.046). The distribution of other pathogenic bacteria such as coagulase-negativeStaphylococci andE. coli was not significantly different between the two groups (P>0.05, respectively). Drug resistance rates ofStaphylococcus (Staphylococcus aureus and coagulase-negativeStaphylococcus) to oxacillin and piperacillin-sulbactam were higher in the screening group than in the no-screening group [82.6% (19/23) vs 52.9% (18/34),χ2=5.302; 78.3% (18/23) vs 47.1% (16/34),χ2=5.549; bothP<0.05], and no vancomycin resistant bacterial strains were found.ConclusionsAntenatal antibiotic exposure may be effective in reducing the occurrence of prematurity, asphyxia,and GBS infection, but it increases the rate of fungal infection, and is not effective in reducing the incidence of complications and mortality or in changing the distribution of the other pathogens in EOS. Rational indications and timing of antenatal antibiotic exposure should be taken into consideration to reduce drug resistance.

During the course of the investigation of the antifungal metabolites against Peronophythora litchii from the fermentation broth of Streptomyces sp SC120, an antibiotics that exhibited the cytotoxicity to CNE 2 was isolated By spectral (UV, IR, HRMS, 1H NMR, 13 C NMR, 1H 1H COSY, HMQC and HMBC) analyses, its structure was identified as pimprinine The cytotoxicity of pimprinine to CNE 2 was reported for the first time

BACKGROUND:Contraction of three-dimensional collagen gels has been used as a model for the contraction which characterizes both normal wound healing and the development of fibrosis in the tissue. Several factors and cytokines,such as tumor necrosis factor alpha (TNF-α), interleukin-1, prostaglandin E2 and insulin have been proved to play important roles in collagen remodeling in vitro as well as serum extravasation during the fibrotic progress.OBJECTIVE: To observe extracellular collagen matrix contraction and apoptosis of fetal lung fibroblasts in TNF-α,interleukin-1, insulin, prostaglandin E2, albumin and globum under three-dimensional culture, and investigate the effects of cytokines, insuin, serum and serum protein on the remodeling and fibrotic formation of lung tissue.DESIGN: A randomized controlled experiment.SETTING: Department of Respiratory Medicine, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out The experiments were carried out in the respiratory laboratory of Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA from August 2005 to January 2006. Human fetal lung fibroblasts (American Type Culture Collection), Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum, insulin, transforming growth factor (TGF) (R&D), prostaglandin E2, type Ⅰ collagen was extracted from rat-tail tendons.METHODS: In order to investigate the effect of initial collagen concentration in the gels on the contractility of fibroblasts,the appropriate amount of collagen was mixed with distilled water, four fold-concentrated DMEM, and cells were suspended so that the mass concentration was 0.75-2.0 g/L, with a physiological ionic strength and the desired cell concentration. In order to investigate the effect of cell number in the gels on the contraction, the cellular concentration fibroblasts in the gels were prepared to 0.2×107-4×107 L-1. The areas of floating gels were measured daily and the contraction was calculated by contrasting the initial size (％ of initial area). Different serum concentrations (0.01％-0.5％)in the medium were prepared, the serum albumin (0.1％) or globulin (0.1％) were added to the serum-free culture medium to observe the gel contraction. TGF (10 mg/L), interleukin-1 (10 mg/L), insulin (1 mg/L) and prostaglandin E2 (0.1 μmol/L) were added to observe the effects of cytokines and insulin on fibroblasts-mediated collagen gel contraction.The DNA content and cellular survivability in gels in collagen were determined.MAIN OUTCOME MEASURES: Effect of lung fibroblasts on collagen contraction with or without the presence of cytokines in three-dimensional culture; Effect of collagen with different concentration on the proliferation and apoptosis.RESULTS: ① Collagen gel contraction showed a dependence on the number of fibroblasts. ② Collagen gel contraction was augmented by increasing serum concentration in culture medium, and albumin increased the contraction dramatically. ③TGF and insulin significantly increased the contraction, whereas prostaglandin E2 and interleukin-1 significantly inhibited the gel contraction. ④ The lower the initial collagen concentration was, the more gel contracted and smaller final size were observed, and cell apoptosis increased.CONCLUSION: During the fibrotic process, fibroblast population migrated into the injured lung tissue may play an important role in the development of pulmonary fibrosis and serum infiltrating into injury lung tissue may play an role in stimulating the fibrotic progress. Infiltrating fluids and edema result in the dilution of the collagen concentration in the pulmonary interstitial which may lead to stronger contraction and serious fibrosis. In the dense fibrosis area, cells were hard to survive. In consequence, the final structure of fibrotic lung could not been changed and lung fibrosis progressed.

Objective To analyze the composition of and drug resistance in bacteria isolated from the lesions of patients with squamous cell carcinoma of the face and head.Methods Lesional tissue or discharges were obtained from 246 patients with squamous cell carcinoma of the face and head,and subjected to conventional bacterial culture.The isolated bacteria were identified by VITEK TWO automated microbiology system.Antimicrobial susceptibility testing was carried out by Kirby-bauer method.WHONET 5.3 software was utilized for statistical analysis.Results Totally,294 bacterial strains were isolated,including 168 Gram-negative bacteria (57.1％)and 126 Gram-positive bacteria(42.9％).The bacterial isolates were predominated by Staphylococcus aureus(21.4％),followed by Escherichia coli(20.4％),Staphylococcus epidermidis(18.4％),Klebsiellapneumoniae(15.4％)and Pseudomonas aeruginosa(9.5％).The prevalence was 40％,26.7％,42.9％ and 55.6％ respectively for extended spectrum β lactamases-producing E.coli and K.pneumoniae,methicillinresistant staphylococcus aureus(MRSA)and methicillin-resistant coagulase-negative S.epidermidis(MRCNS)respectively.P.aeruginosa,E.coli and K.pneumoniae were highly susceptible to imipenem and meropenem,and favorably sensitive to β-lactam and β-1actamase inhibitor combination.No resistance was observed for vancomycin,teicoplanin or linezolid in staphylococci.Conclusions The bacterial isolates from squamous cell carcinoma tissue on the head and neck are predominated by conditional pathogenic bacteria,and the proportion of Gram-negative bacteria is higher than that of Gram-positive bacteria.These isolates seem to be highly resistant to common antibiotics.