摘要:目的: 金黄色葡萄球菌(金葡菌)的感染近年来已成为医院内的主要致病菌,而其耐药性也呈逐渐升高的趋势,为了解该菌在我院的感染和耐药情况,为临床合理使用抗生素提供科学依据。 方法 : 用经典生理生化鉴定方法,对各种临床标本主要来源于痰液和各种伤口脓液标本分离到的102株金葡菌进行生物学特性及药敏试验。 结果 : 从我们医院2007年5月至2009年8月所分离出来的102株金葡菌中青霉素耐药性8923%,氨苄青霉素耐药率为9385%,没有发现万古霉素耐药菌。 结论 : 除万古霉素外,耐药率较低的依次是利福平、苯唑青霉素、环丙沙星、呋喃妥因、阿米卡星、磺胺甲基异恶唑、红霉素,而青霉素G、氨苄青霉素、四环素耐药性情况非常严重,并且多重耐药,耐药性强,应引起临床的高度重视。Abstract: Objective: To analyze the bionomics and antimicrobial susceptibility of staphylococcus aureus, which was the main pathogenic bacterium with high drug tolerance in our hospital, in order to provide the rational use of antibiotics. Methods : Samples of one hundred and two staphylococcus aureus cases from sputamentum and pus were evaluated by classic physiology and biochemistry methods to test the bionomics and antimicrobial susceptibility. Results : The drug resistance rate to penicillin, penbritin and vancomycin was 8923%, 9385% and 0, separately. Conclusion : Besides vancomycin, the drug resistance rate of rifampicin, oxazocilline, ciprofloxacin, furadantin, amikacin, sulfamethoxazole and sulfamethoxazole increased one by one. The resistance to penicillin G, penbritin and tetracycline was serious, including multidrug resistant, which should be paid highly attention.
Objective To explore a method to isolate, culture and multiplicate the placentaderived mesenchymal stem cells (PMSCs) and the bone marrow-derived mesenchymal stem cells (BMSCs) of rabbit，and to compare their biological characteristics. Methods PMSCs were isolated from placenta of 1fetation rabbitby Percoll density gradient centrifuge and cultured in vitro. BMSCs were isolated from hindlimb bone marrow blood of 1 new born rabbit by direct plates culturemethod. The 3rd passage PMSCs and BMSCs were observed by inverted phase contrast microscope. The stem cell marker (CD44, CD105, CD34 and CD40L) were examined by immunohistochemistry. The 2nd passage PMSCs and BMSCs were co-cultured with biomaterials，(1.0-1.5)×106 cells in one biomaterial， and then observed by aematoxylinstaining after 5 days,and by SEM after 3 days and 8 days. Results PMSCs and BMSCs were both uniformly spondle-shaped in appearance and showed active proliferative capacity. The proliferative ability of PMSCs were quite b and declined with passages. After cultured 10 passages in vitro, its growthslowed. Both PMSCs and BMSCs expressed CD44 and CD105,but did not express CD34 and CD40L immunoreactivity. PMSCs and BMSCs poliferated and adhered to the surface of biomaterials, and cell formed clumps and network; the cells proliferation and the matrix were seen in the pore after 5 days of culture. The observation ofSEM showed that many cells adhered to the biomaterials with spindle-shape and polygon after 3 days; and that PMSCs and BMSCs grew,arranged in layers andsecreted many matrices; the reticular collagen formed arround cells after 8 days. Conclusion PMSCs and BMSCs have similar biological characteristics and PMSCs can be served as excellent seedingcells for tissue engineering.
【Abstract】 Objective To explore an effective method to cultivate esophageal mucosa epithel ial cells (EMECs)of canine in vitro, and to observe the biological characteristics of EMECs growing on SIS in order to provide an experimental basis for esophagus tissue engineering. Methods Esophageal tissues were obtained from five healthy dogs aged 2 to 5 weeks under sterile conditions. The primary EMECs were cultivated with defined keratinocyte serum free medium (DKSFM) containing 6% FBS. The morphological characteristics and the growth curve of EMECs of the 2nd generation were observed for 1 to 5 days. The expressions of the EMECs marker (cytokeratin 19, CK-19) were examined by immunocytochemistry. The 2nd generation of EMECs was seeded on SIS and observed by HE staining, immunohistochemical staining, and SEM for 4 and 8 days. Results The primary culture of canine EMECs arranged l ike slabstone. Immunohistochemical staining of CK-19 of the2nd generation EMECs showed positive broadly. The cells growth reached the peak level at 2 days by MTT method. E MECs werepolygon in shape and arranged l ike slabstone, and formed a single layer on the surface of SIS. The cells were contact ed closely with each other for 4 days. Eight days later, 2 to 3 layers stratified structure was formed. Lots of EMECs were grown on SIS, andshowed laminate arrangement. Conclusion With mixed enzymatic digestion, the culture of EMECs in DKSFM containing 6 %FBS is a simple and feasible method. SIS shows good biocompatibil ity and can be used as a good scaffold material in th e tissue engineered esophagus.
ObjectiveTo summary the relationship between CD147 and the occurrence and development of hepatocellular carcinoma, and its roles in clinical diagnosis and treatment. MethodReferring to the related literatures in recent years at home and abroad, the concept of CD147, and its relationship with the occurrence, development, invasion, and metastasis of hepatocellular carcinoma were reviewed. ResultsCD147 plays a key role in the development, progress, invasion, and metastasis of hepatocellular carcinoma. CD147 can be used as a long-term outcome predictor for hepatocellular carcinoma patients and also began to show its in hepatocellular carcinoma target therapy. ConclusionThere are numerous studies about the relationship between CD147 and hepatocellular carcinoma, but still exists some problems to be further studied.
Following the peritendon was removed by means of microsurgical technique, the tenocyte was isolated from the human embryonic tendons by digesting it with trypsin and collagenase. These cells were all stored in frozen condition until they were cultured by F12 culture fluid added with 20% FBS to the 15th generation.These cells were able to grow adhering to the wall and stop growing with contact inhibition. The time of cellsgroup duplication was 4 days, which was similar to the peak time of its mitosis. The number of its chromosome group 2n=46 was 87.5-91.0%. The optimal conditions for tendon cell culture in vitro were investigated, and it was found that after they were reaminated and subcultured the frozen storage didn’t influence their growth, morphology, genetic characteristics. In our research we detected the content generation cells and found the cultured human embryonic tenocyte had same ability never changed with the cells subcultured. We also disscussed the future of tenocyte-a biomaterial in the field of artificial implant.
OBJECTIVE This paper was to study the biological characteristics of the transformed human embryonic tendon cells, the relation between cell growth and culture conditions, and to compare these features with that of human embryonic tendon cells. METHODS The pts A58H plasmid had successfully used to transform a tendon cell line from human embryo in our past work. The human embryonic tendon cells and the transformed human embryonic tendon cells were cultured in vitro. In different culture conditions, the growth curve were drawn respectively. Population dependence and proliferation capability of the cells were investigated through plate cloning test and soft agar culture. The collagen secreted by cells was identified by immunohistochemical method. RESULTS In routine culture condition, the growth properties of the human embryonic tendon cell and transformed cells were almost identical. The growth properties of the transformed cells were not changed when the cells were frozen storage. There were changes of growth characteristics of the transformed cells when the culture temperature was changed. The transformed cells could subcultured continually and permanently. The proliferation capability of the transformed cells were ber than that of the human embryonic tendon cells. Moreover, the growth of the transformed cells was serum-dependent, and the phenomenon of contact inhibition was observed. The transformed cells were not able to grow on soft agar culture. They had the capacity of secreting collagen type I. CONCLUSION The transformed human embryonic tendon cells could be subcultured continually and permanently, and their growth could be controlled by changing their culture conditions and they had no malignant tendency in biological characteristics. They could be taken as an ideal experimental material for tendon engineering.
【Abstract】 Objective To observe the effects of Angelica dahurica extracts on the biological characteristics of human keratinocytes (KC) in vitro and to explore the possible mechanism in promoting wound healing. Methods HaCaT cells of passage 5 from KC were used during the experiment. Different concentrations (5 × 10-2, 5 × 10-3, 5 × 10-4, and 5 × 10-5 g/L) of Angelica dahurica extracts, which was obtained by 95% ethanol from Angelica dahurica raw material, were prepared by DMEM containing 0.25% fetal bovine serum (FBS). After the extracts at different concentrations were respectively used for KC culture for 5 days, the cell proliferation activities were detected by MTT, and DMEM containing 0.25% FBS served as the negative control. According to the cell proliferation activity, the optimal concentration was determined. KC was further treated with Angelica dahurica extracts of the optimal concentration (experimental group) or with DMEM containing 0.25% FBS (control group) for 48 hours. The cell cycle was tested by flow cytometry. Cyclin D1 and Caspase-3 mRNA levels were also detected by real-time fluorescent quantitative PCR technique. Results Angelica dahurica extracts at concentrations of 5 × 10-4, 5 × 10-3,and 5 × 10-2 g/L could significantly enhance KC proliferation, showing significant differences in absorbance (A) values compared with that of control group (P lt; 0.05) with an optimal concentration of 5 × 10-3 g/L. At this concentration, an increased percentage of S and G2/M phase cells and a decreased percentage of G0/G1 phase cells were detected, showing significant differences when compared with control group (P lt; 0.05). Real-time fluorescent quantitative PCR revealed that the cyclin D1 and Caspase-3 mRNA levels of experimental group was significantly down-regulated, showing significant differences when compared with control group (P lt; 0.05). Conclusion Angelica dahurica extracts can promote the proliferation of KC, accelerate the cell cycle of KC by down-regulating mRNA expressions of cyclin D1, and inhibit apoptosis by down-regulating mRNA expressions of Caspase-3. These effects might enhance the process of wound healing by expediting the process of epithelization.
Objective To investigate the biological and biomechanical characteristics of acellular porcine aortic valve with dye mediated photo oxidation so that a new and better bioprosthetic valve materials can be obtained. Methods Thirty porcine aortic valves were divided into three groups with random number table. Acellular valves (n=10) were stabilized by dye mediated photo oxidation in dye mediated photo oxidation group; acellular valves (n=10) were stabilized by glutaraldehyde in glutaraldehyde group; and acellular valves (n=10) were acellularized only in acellular valves group. Thickness, appearance, histology, water content, shrinkage temperature, breaking strength and soluble protein level of acellular porcine aortic in three groups were tested respectively. Results There were light blue, soft, flexible and unshrinking valves in dye mediated photo oxidation group. Compared to valves in glutaraldehyde group, valves in dye mediated photo oxidation group had lighter thickness（0.26±0.09mm vs. 0.38±0.08mm，Plt;0.05）, more water content（86.30%±4.03% vs. 71.10%±3.23%，Plt;0.05）, and lower shrinkage temperature （76.30±0.70℃ vs. 87.70±0.30℃，Plt;0.05）; while these indexes had no statistically significant differences compared to those in acellular valves group. At the same time, compared to valves in acellular valves group, valves in dye mediated photo oxidation group had more breaking strength（17.33±2.65 mPa vs. 9.11±0.95 mPa，Plt;0.05） and lower soluble protein level（0.039%±0.013% vs. 0.107%±0.024%，Plt;0.05）; while these indexes had no statistically significant differences compared to those in glutaraldehyde group. Conclusion Acellular porcine aortic valve stabilized by dye mediated photo oxidation has nice biological and biomechanical characteristics.
ObjectiveTo review the current progresses in purification strategies, biological characters, and the uses in tissue engineering of urine-derived stem cells (USCs). MethodsRecent relevant publications on the USCs were extensively reviewed, analyzed, and summarized. ResultsUSCs, usually isolated by adherence screening method, are a population of mesenchymal stem cells (MSCs)-like somatic stem cells possessing robust self-renew and multi-potential differentiation ability. Combined with using appropriate biomaterials and biological molecules, USCs can be used as a good cell source for tissue engineering. ConclusionAn alluring prospect exists on the USCs-related research. Further studies are required to investigate the origin, individual differences, and the therapeutic values of USCs.
Objective To summarize the research progress of biological characteristics and advantages of Wharton’s jelly-mesenchymal stem cells (WJ-MSCs). Methods The related l iterature on the biological characteristics of WJ-MSCs,umbil ical cord blood MSCs (UBMSCs) and bone marrow MSCs (BMSCs) was extensively reviewed and analyzed. Results A large number of MSCs which are able to self-repl icate, self-renew and have high prol iferation and multipotent differentiation can be isolated from the Wharton’s jelly of umbil ical cord. WJ-MSCs have many advantages in isolation time, isolation efficience, expansion time, passage capacity, expansion capacity when compared with UBMSCs and BMSCs. Conclusion WJ- MSCs have numerous advantages of convenient and abundant sources, relatively pure, non-ethical issues, and so on, which can be used for cell transplant therapy, gene therapy, and the ideal seed cells of building tissue engineered organ, so they provide new ideas for tissue regeneration repair and reconstruction.