1.Bacterial Spectra Investigation of Blood Culture and Clinical Analysis of Resistance of Pathogens
Yunzhuo CHU ; Hua NIAN ; Yuxin DENG ; Jinming OUYANG ; Qian WANG
Chinese Journal of Nosocomiology 1994;0(04):-
OBJECTIVE To offer the clinical physician the basis of optimal application of antibiotic,we have investigated the variation of antibiotic resistance and the bacterial spectra in the blood culture.METHODS Blood was cultured in BACTEC9120 of BD.The clinical isolates were identified by API and VITEK-2 of Bio-Merieux of France.Antibiotic susceptive test was done by Kirby-Bauer method and the result which was analyzed by WHONET5.3 and SPSS11.5 software was determined by the NCCLS standard of 2005′s edition.RESULTS Organisms were isolated from the blood specimen of 1468 patients,and there were 743 strains of Gram-positive cocci accounted for 50.7%,565 strains of Gram-negative bacilli accounted for 38.5%.Ninety three strains of fungi accounted for 6.3%.We analyzed the drug-susceptive result of Staphylococcus,Escherichia coli,and Klebsiella pneumoniae during five years,and found that all the antibacterial drug lost efficacy in some degree,except that the sensitivity of the staphylococci to vancomycin was 100%.CONCLUSIONS Gram-positive cocci are the main bacteria in blood culture,the species from which are diversified,and the rate of the drug resistance of some bacteria is high.It indicated that doctors should take more blood culture and monitor the bacteria drug resistant for the data of etiology,so that they can utilize antibiotic more reasonably.
2.In vitro drug release rule of basic fibroblast growth factor-poly(lactic-co-glycolic-acid) copolymer microspheres:Promotion of the venous flap survival in rabbits?
Hongju XIE ; Ming LI ; Ying DENG ; Nian CHEN
Chinese Journal of Tissue Engineering Research 2007;0(51):-
BACKGROUND:Compared with normal physiological flap,main advantage of venous skin flap is that it throws off the limitation of arterial vascular territory on donor site and recipient site of traditional axial skin flap.However,its survival rate is unstable.OBJECTIVE:To explore effects of basic fibroblast growth factor (bFGF)-[poly(lactic-co-glycolic-acid)] (PLGA)-sustained release microspheres on the survival of rabbit venous flaps.DESIGN,TIME AND SETTING:A randomized controlled animal study was performed at the University of South China from May to October 2008.MATERIALS:A total of 24 healthy New Zealand rabbits were equally randomly assigned to bFGF-PLGA sustained release microsphere,blank microsphere and blank control groups.METHODS:The formulation of bFGF microspheres was optimized by orthogonal design.bFGF-PLGA microspheres were prepared by optimized method.Lateral abdominal wall skin flap was created in rabbits from 3 groups.Five days before operation,28.85 g/L bFGF-PLGA microspheres 3 mL (containing bFGF 20 ?g) was intradermally injected into rabbits from the bFGF-PLGA sustained release microsphere group.An equal volume of blank microsphere + bFGF was injected in the rabbits of the blank microsphere group.Rabbits from the blank control group were infused with the same volume of saline.MAIN OUTCOME MEASURES:Morphology and particle distribution of bFGF-PLGA microspheres,drug loading volume,encapsulation efficiency,in vitro drug release characteristics were measured.After seven days,the survival area of skin was determined.Rabbit skin samples received CD34+ immunohistochemical staining to detect the expression of CD34+ and average number of blood vessels.RESULTS:The bFGF microsphere prepared based on optimized formulation exhibited well-defined properties,with the even and uniform sphere in appearance,regular particles without adhesion,about 98% of particles with a size distribution between 12.50 to 43.49 ?m,with a mean particle size of 26.93 ?m and size span of (0.611 ? 6.60).The drug loading volume and encapsulation efficiency of bFGF microsphere reached [(23.11?0.44)?10-3]% and (86.51?0.83)%,respectively.In the burst release phase,the rate of in vitro drug release amounted to 27.78%,but rose to 81.56% accumulatively 30 days later.The in vitro drug release of bFGF microsphere corresponded with Higuichi equation (r=0.997).The sustained-release microspheres,blank microspheres and normal saline group,the average survival of the flap and the average number of blood vessels were similar (P=0.597,P=0.336),but still significantly lower than the bFGF-PLGA sustained release microsphere group (P=0.000).Results of immunohistochemical staining revealed that bFGF-PLGA promoted blood supple between flap and surroundings,improved flap survival and abundant CD34+ expression.CONCLUSION:bFGF microsphere with good morphology,high drug loading volume and encapsulation efficiency can be obtained using W/O/W multiple emulsion evaporation method.The bFGF microsphere can promote the survival of rabbit venous flaps through a long period due to sustained release of bFGF.
3.In vitro drug release rule of basic fibroblast growth factor-poly(lactic-co-glycolic-acid) copolymer microspheres: Promotion of the venous flap survival in rabbits?
Hongju XIE ; Ming LI ; Ying DENG ; Nian CHEN
Chinese Journal of Tissue Engineering Research 2009;13(51):10039-10044
BACKGROUND: Compared with normal physiological flap, main advantage of venous skin flap is that it throws off the limitation of arterial vascular territory on donor site and recipient site of traditional axial skin flap. However, its survival rate is unstable. OBJECTIVE: To explore effects of basic fibroblast growth factor (bFGF)-poly(lactic-co-glycolic-acid)] (PLGA)-sustained release microspheres on the survival of rabbit venous flaps.DESIGN, TIME AND SETTING: A randomized controlled animal study was performed at the University of South China from May to October 2008.MATERIALS: A total of 24 healthy New Zealand rabbits were equally randomly assigned to bFGF-PLGA sustained release microsphere, blank microsphere and blank control groups.METHODS: The formulation of bFGF microspheres was optimized by orthogonal design. bFGF-PLGA microspheres were prepared by optimized method. Lateral abdominal wall skin flap was created in rabbits from 3 groups. Five days before operation, 28.85 g/L bFGF-PLGA microspheres 3 mL (containing bFGF 20 μg) was intradermally injected into rabbits from the bFGF-PLGA sustained release microsphere group. An equal volume of blank microsphere + bFGF was injected in the rabbits of the blank microsphere group. Rabbits from the blank control group were infused with the same volume of saline. MAIN OUTCOME MEASURES: Morphology and particle distribution of bFGF-PLGA microspheres, drug loading volume, encapsulation efficiency, in vitro drug release characteristics were measured. After seven days, the survival area of skin was determined. Rabbit skin samples received CD34~+ immunohistochemical staining to detect the expression of CD34~+ and average number of blood vessels. RESULTS: The bFGF microsphere prepared based on optimized formulation exhibited well-defined properties, with the even and uniform sphere in appearance, regular particles without adhesion, about 98% of particles with a size distribution between 12.50 to 43.49 μm, with a mean particle size of 26.93μm and size span of (0.611 ± 6.60). The drug loading volume and encapsulation efficiency of bFGF microsphere reached [(23.11 ±0.44 )x10~3]% and (86.51±0.83)%, respectively. In the burst release phase, the rate of in vitro drug release amounted to 27.78%, but rose to 81.56% accumulatively 30 days later. The in vitro drug release of bFGF microsphere corresponded with Higuichi equation (r= 0.997). The sustained-release microspheres, blank microspheres and normal saline group, the average survival of the flap and the average number of blood vessels were similar (P=0.597, P=0.336), but still significantly lower than the bFGF-PLGA sustained release microsphere group (P=0.000). Results of immunohistochemical staining revealed that bFGF-PLGA promoted blood supple between flap and surroundings, improved flap survival and abundant CD34~+ expression. CONCLUSION: bFGF microsphere with good morphology, high drug loading volume and encapsulation efficiency can be obtained using W/O/W multiple emulsion evaporation method. The bFGF microsphere can promote the survival of rabbit venous flaps through a long period due to sustained release of bFGF.
4.Hybrid aortic endovascular repair with one stage supra-aortic branch or tliac artery revascularization
Yuehong ZHENG ; Nian CAI ; Hongru DENG ; Changyu GUO ; Furtado RUI
Chinese Journal of General Surgery 2009;24(11):915-918
Objective Repair of aortic arch aneurysm is technically demanding and usually requiring complex circulatory management. Operative morbidity and mortality may be prohibitive with traditional surgical intervention. We described our experience with 5 hybrid endovascular procedure for aorta repair with different kinds of bypass followed by concomitant placement of stent graft in the aorta. Methods We retrospectively reviewed the clinical data of 5 consecutive patients presenting with aortic aneurysm or dissection from 2007 to 2008 treated by the hybrid aorta repair. Complete surgical rerouting of the supra-aortic vessels was followed by endovascular repair of aortic arch aneurysm with a Zenith TX2 stent graft. Hybrid left carotid-subclavian bypass with Zenith stent graft deployment covering the ostium of the LSA was performed in a Debakey type Ⅲ aortic dissection case. Procedures were successfully completed with exclusion of the aortic aneurysm. All stent grafts were deployed retrograde from the femoral artery in these patients. Results Technical success with complete aneurysmal exclusion was achieved in all patients (100%). At a follow-up period of 2-10 months, there was no incidence of endoleak. Documented perioperative neurelogic events did not occurred in all patients. Postoperatively one patient suffered from ARDS and cardiac failure and recovered. One patient died of myocardial infarction. Conclusions Hybrid arch repair provides an alternative to patients otherwise considered prohibitively high risk for traditional open arch and thoracoabdominal aorta repair.
5.Transplantation of autologous costal cartilage to repair post-traumatic saddle nose deformity in 21 cases
Nian CHEN ; Ying DENG ; Ming LI ; Li ZENG ; Hongju XIE
Chinese Journal of Tissue Engineering Research 2010;14(18):3363-3366
BACKGROUND: The soft tissues would shrink with nasal framework collapse following surgical trauma, which cause aseptic inflammation, lead to parts of cartilage resorption. Accordingly, long-term saddle nose deformity usually accompanied by short nasal columella. Complications such as skin perforation or ulceration would appear if corrected the deformity using medical silicone rubber with large tension.OBJECTIVE: To explore the effectiveness of repairing post-traumatic saddle nose deformity with autologous costal cartilage transplantation.METHODS: A total of 21 cases with post-traumatic saddle nose deformity accompanied by short nasal columella were selected, including 6 males and 15 females, aged 16 45 years. All of the cases had trauma history and agreed with the treatment. The costal cartilage was obtained from the seventh rib and formed to babylon weeping willow leaf shape and columella nasi stent to repair post-traumatic saddle nose deformity. The "V-Y" progradation suture was used in the philtrum introcession and the botton of nasal columella to extend the nasal columella. The recovery of saddle nose deformity after transplantation, discharge of transplanted cartilage, as well as the incision scar status was observed.RESULTS AND CONCLUSION: The results were satisfactory and there were no complications after transplantation. All the cases were followed up from 6 months to 2 years. No case suffered costal cartilage grafts discharge or chondral deformation. The scar was little at the bottom of nasal columella. It is an ideal method for repairing post-traumatic saddle nose deformity using transplantation of autologous costal cartilage with "V-Y" progradation suture.
7.Research progress of associated risk factors in intervertebral disc degeneration
Chensheng QIU ; Nian DENG ; Hongfei XIANG ; Yongsheng ZHAO ; Bohua CHEN
Chinese Journal of Orthopaedics 2021;41(10):654-659
Low back pain is an important cause of disability worldwide. It has a high incidence rate and brings a huge burden to families and society. Intervertebral disc degeneration (IDD) is one of the leading factors causing low back pain and the pathological basis of degenerative disc diseases, such as intervertebral disc herniation and spinal stenosis. However, the etiology of IDD is complex, and the risk factors and specific mechanisms behind remain unclear. Some controversial views have also been observed. Surgery is often considered for patients with severe intervertebral disc diseases, but there is no effective treatment for IDD at the early and middle stages. It will be of great significance to in-depth explore the molecular biological mechanisms and related risk factors, which can bring benefits to the prevention, accurate diagnosis, early treatment, and rehabilitation of degenerative disc diseases. Refer to the literatures published in the past ten years, this paper describes the latest research progress on risk factors related to IDD in terms of aging, genetics, mechanical loading, low-grade infection, biological rhythms, smoking, metabolic disease, estrogen, and nutrition. The results show that IDD is affected by multiple risk factors. These factors can interact with each other, and lead to death, phenotypic transformation, and metabolic disorder of disc cells, leading to a reduction of extracellular matrix and an unbalanced microenvironment and eventually loss of structural integrity of intervertebral disc tissue and IDD. A good body clock, a controlled weight, an appropriate blood glucose level, adequate nutrition, no smoking, a good hormone level, moderate exercise, avoiding injury, and strict aseptic techniques in the clinic will bring benefits to the progress of IDD.
8.Nerve growth factor potentiates bone morphogenetic protein-9-induced osteogenic differentiation in mouse embryonic fibroblasts
Changjun PI ; Xiang ZOU ; Ranxi ZHANG ; Nian WU ; Baicheng HE ; Zhongliang DENG ; Liang CHEN
Chinese Journal of Trauma 2016;32(2):171-175
Objective To investigate the effect of nerve growth factor (NGF) on osteogenesis induced by bone morphogenetic protein-9 (BMP-9) in mouse embryonic fibroblasts (MEFs).Methods MEFs were respectively transfected with adenovirus-mediated NGF (NGF group), BMP-9 (BMP-9 group) and NGF + BMP-9 (combined group) and green fluorescence protein (GFP) (control group).Cytochemical staining was used to test the activity of alkaline phosphatase (ALP) 3 d and 5 d after treatment.Level of osteopontin (OPN) mRNA was detected by RT-PCR 9 d after treatment.Level of OPN protein was assayed by Western blot and immunocytochemistry 9 d after treatment.Mineralization was detected by Alizarin red staining 14 d after treatment.Results ALP activity in MEFs was elevated in BMP-9 group rather than in NGF group, but a significant increase in ALP activity was noted in combined group.In control group, BMP-9 group, NGF group and combined group, level of OPN mRNA was 0.92 ± 0.03, 1.28 ± 0.04, 0.94 ± 0.03 and 1.62 ± 0.04 respectively (F =214.60, P < 0.01);level of OPN protein was 0.60 ± 0.05, 0.84 ± 0.03, 0.53 ± 0.05 and 1.27 ± 0.05 respectively (F =162.5, P < 0.01).In comparison, OPN mRNA and protein were significantly up-regulated in combined group than in BMP-9 group (t =10.569 and 11.778,P < 0.05).In control group, BMP-9 group, NGF group and combined group, relative density of OPN protein was 3.63 ±0.17, 6.27 ±0.30, 3.86 ±0.18 and 10.16 ±0.18respectively (F =602.6, P < 0.01), with a significant higher level in combined group than in BMP-9 group (t =22.280, P < 0.05).Level of mineralization was significantly higher in combined group than in BMP-9 or NGF group.Conclusion NGF can potentiate the osteogenesis induced by BMP-9 in MEFs.
9.Construction and characterization of recombinant pseudovirus particles carrying specific genomic RNA of St.Louis encephalitis virus
Nana ZHANG ; Yongqiang DENG ; Qinggong NIAN ; Xiaoping KANG ; Yinhui YANG ; Chengfeng QIN
Military Medical Sciences 2017;41(3):194-198
Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.
10.Effect of trehalose on survival rate for fat cells after cryopreservation
Ying DENG ; Shaoqian LIU ; Hongju XIE ; Fangfang TANG ; Ming LI ; Nian CHEN
Journal of Central South University(Medical Sciences) 2017;42(5):507-510
Objective:To explore effects of trehalose as a cryopreserve agent on survival rate of fatty tissue after cryopreservation.Methods:The liposuction was used on the abdomen of adult female.After centrifugation and purification,adipose was randomized into the following three groups,the trehalose group,the fetal bovine serum (FBS)+ 10%DMSO group and the physiological saline group.The specimens were cryopreserved at-196 ℃ for 3 months and then the HE staining,glucose transfer method and CK method were used to detect the cell survival rate in each group.Results:The activity of adipose in the trehalose group and FBS+10%DMSO group adipose was higher than that in the physiological saline group (P<0.05);while there was no significant difference between the trehalose group and FBS+10%DMSO group (P>0.05).Conclusion:As cryoprotectant,trehalose could keep fat cell viability,and adipose tissue can be used for clinical transplantation after 3 months' freezing.