1.Isolation of Staphylococcus aureus from Detention Needles of Burn Patients and Analysis of Its Antibiotic Resistance
Ping ZHOU ; Mo SHEN ; Xuezhong HUANG ; Peipei LIN ; Shaoguan HUANG
Chinese Journal of Nosocomiology 1994;0(01):-
OBJECTIVE To study the detectable rate and the antibiotic resistance of Staphylococcus aureus from detention needles of burn patients,and provide instruction of clinic reasonably using antibiotics and clinical treatment.METHODS Totally 175 specimens detention needles of from burn patients were collected to isolate pathogenic organisms.Identification of bacterial strains and susceptibility tests were performed by ATB system.RESULTS S.aureus isolated from the detention needles of burn patients was the predominant pathogen,accounted for 60.3%,and all of them were MRSA.Among these flora,all were resistant to penicillins,oxacillin and gentamicin and more than 50.0% were resistant to lincomycin,tetracyc1ine,rifampicin and so on,2.3% were resistant to minocycline and teicoplanin,and none was resistant to vancomycin, fusidic acid,and quinupristin-dalfopristin.CONCLUSIONS S.aureus isolated from the detention needles of burn patients is the predominant pathogen, which is resistant to many antibiotics.So we should identify the bacterial strains and give susceptibility tests of detention needles from burn patients promptly,in order to provide best instruction on clinic reasonably using antibiotics and slow the development of the resistant bacterial flora.
2.Clinical value of serum procalcitonin in the diagnosis of bacterial infections in the critical patients
Yan SHEN ; Kuibin LIN ; Xiuchun HUANG ; Yuezhao CHEN ; Xiaoli CHEN
Chinese Journal of Primary Medicine and Pharmacy 2009;16(6):975-976
Objective To evaluate the clinical value of serum procalcitonin (PCT) in the diagnosis of bacte-rial infections in the critically ill patients. Methods A loud d 100 cases of critical patients were divided into bacteri-al infection group(56 cases) and non-bacterial infection group(44 cases). Serum PCT was measured by immunolu-minometric assay. Results The concentration of PCT in bacterial infection group(21.54 ±5.72) μg/L, was signifi-cantly higher than non-bacterial infection group (11. 95±4. 58)μg/L (t =2.291,P<0.05);The APACHE Ⅲ score of bacterial infection group(62. 44 ± 19. 55) cent was significantly higher than non-bacterial infection group(44. 56 ± 25. 88) cent(t = 2. 195 ,P < 0. 05). The concentration of PCT of 1.0μg/L and 2. 0 p.g/L compared to the former sensitivity (96. 5) % higher than the latter (55.2) % (X2 = 3. 94, P < 0. 05), the former specificity (41.7) % lower than the latter (95.8)% (X2 = 4. 02 ,P < 0. 05);1.5μg/L as a positive standard Youden index and the Agreement (83.0 % ,0. 65), were significantly higher than 1 μg/L and 2 μg/L(71.7%, 0. 38 ;73.6% ,0. 51) (X2 = 3.84, X2=3. 90,X2 = 3.992 = 3.91 ,P < 0. 05);The concentration of PCT in death group (38. 9 ±12. 6)μg/L was signifi-canfly higher than the survival group(11.8± 8. 3) μg/L(t =2. 398 ,P <0. 05). Conclusion Serum PCT has clinical values in the diagnosis and therapy of bacterial infections in the critical patients.
3.Protective effects of ulinastatin on phosgene-induced acute lung injury and the expression of tumor necrosis factor-alpha
Wenbin HUANG ; Jie SHEN ; Lin ZHANG ; Daikun HE
Chinese Journal of Emergency Medicine 2010;19(1):37-42
Objective To observe the effects of ulinastatin on the expressions of tumor necrosis factor-α (TNF-α) in the lung tissues of rats with acute lung injmy induced by phosgene, and to explore the mechanism of ulinastafin in treating acute lung injury. Method Sixty-four clean grade healthy male SD rots were randomly divided(random number) into eight groups with eight in each group. Group A1 in which rats were exposed to air. Group A2 in which rots wereexposed to air and treated with saline. Group A3 in which rats were exposed to air and treated with dexamethasone. Group A4 in which mrs were exposed to air and treated with ulinastatin. Group B1 in which rots were exposed to phosgene without treatment. Group B2 in which rats were exposed to phosgene and treated with saline. Group B3 in which rats were exposed to phosgene and treated with dexamethasone. Group B4 in which rats were exposed to phosgene and treated with ulinastatin. The expressions of TNF-α in the lung tissues were measured by using immunohistocheistry. Lung tissues were observed grossly and under 200-fold light microscope to identify the positive expressions in kytoplasm. Results In group B1 and group B2, the wet weight, dry weight and wet/dry weight ratio og right lower lobe of lung were higher thai those in group A1 and group A4( P <0.01 ), and those in group B4 and group B3 were significantly lower than those in group B1(P<0.01 ), but still higher than those in group A1 and A4(P<0.01). The gross observation suggested that the surfaces of lung tissues in group A1and group A2 were slick and rose pink without congestion, dropsy or infarction; the surfaces of lung tissue in groups B1 and B2 appeared with congestion, dropsy and many petechia. The surfaces of lung tissue in groups B3 and B4 were similar to those in groups B1 and B2. Under the light microscope, the structure, of lungs in groups A1 and A2 were clear without congestion, effusion or inflammatory cell infiltration. In groups B1 and B2, the engorgement of lung capillary vessels, congestion and tiny thrombosis were found and there abundant edematous fluid and inflammatory cell infiltration in lung stroma and alveolus with focal pulmonary atelectasis in some lung tissue section. The tissues in groups B3 and B4 showed congestion, dropsy, tiny thrombosis and inflammatory cell infiltration, but these changes were slighter than those in groups B1 and B2. The expressions of TNF-α in groups B1, B2, B3, and B4 were significantly higher thanthose in groups A1 and A2( P < 0.01 ), but the expressions of TNF-α IN group B4 was lower than that in groups B1 and B2(P<0.01). Conclusions Ulinastatin could lessen the lung injury by reducing the expressions of pro-inflammatory cytokines such as TNF-α.
4.Metal implants for treatment of unstable pelvic fracture
Zhijin LIN ; Liehu CAO ; Feng SHEN ; Jianming HUANG ; Yifan KANG
Chinese Journal of Tissue Engineering Research 2010;14(9):1665-1668
BACKGROUND:It is controversial to treat sacroiliac joint fracture.Some scholars advocated expectant treatment,and some others advocated surgery therapy.Pelvic stability was responsible for the scheme selection.Sacroiliac joint fracture-dislocation destroys pelvic stability,which easily induces instability and bone nonunion,resulting in sacroiliac joint pain,unequal size of lower limbs,sitting pain and dysfunction.Thus,prognosis of mental implant is significantly better than expectant treatment in treatment of unstable pelvic fracture.OBJECTIVE:To summarize the treatment of unstable pelvic fractures and their clinical application using various internal and external fixation of metal implants.METHODS:The computer-based research was done in Pubmed database (http://www.ncbi.nlm.nih.gov/PubMed) and Wanfang database (http://www.wanfangdata.com.cn) for articles published from January 1991 to December 2009 with the key words of "Pelvic fractures,instability,surgical treatment" by the first author.A total of 115 articles were retrieved,and those concerning characteristics and clinical application of implants in the treatment of unstable pelvic fracture.Articles addressing old and repetitive contents were excluded.Literatures of the same fields published in recent years or in authorized journals were selected.Finally,30 articles were included.RESULTS AND CONCLUSION:Sacroiliac joint fracture-dislocation is a severe,high-energy trauma,has been paid great attention in the therapy,particularly in unstable sacroiliac joint fracture-dislocation.A stable type of fracture and dislocation of the sacroiliac joint received a conservative treatment of unstable sacroiliac joint fracture-dislocation appropriate line of external fixation,internal fixation for reconstruction of pelvic stability and internal fixation treatment varied,but the therapeutic effect of internal fixation needs to be improved.What are bio-mechanical characteristics of various internal fixation methods,and how the timing of weight-bearing activities following various internal fixations require further basic and clinical studies.An unstable sacroiliac joint fracture-dislocation fixation has many ways,including the anterior and posterior fixed-fixed.Minim ally invasive therapy such as posterior CT guided sacroiliac joint lag screw is the developmental trend.
5.Effect of low-intensity pulsed ultrasound on cell-free demineralized bone matrix co-cultured with rabbit cartilage cells and bone marrow mesenchymal stem cells in vitro
Zhijin LIN ; Hao TANG ; Feng SHEN ; Jianming HUANG ; Yifan KANG
Chinese Journal of Tissue Engineering Research 2009;13(47):9217-9223
BACKGROUND: Using low-intensity pulsed ultrasound (LIPU) to promote the repair of articular cartilage injury is very common,and we also have more options to choose the cytoskeleton, but the application conditions of LIPU and the appropriate cytoskeleton have not reached any consensus yet.OBJECTIVE: To investigate the feasibility of establishing tissue-engineered cartilage by cell-free allograft demineralized bone matrix (CFDBM) co-cultured with rabbit cartilage cells and bone marrow mesenchymal stem cells (BMSCs) in vitro, and to investigate the effect of LIPU on the cells in CFDBM.DESIGN, TIME AND SETTING: Multiple sample observation was performed at the Institute of Biomedical Engineering, Second Military Medical University of Chinese PLAfrom May to August 2009.MATERIALS: The CFDBM was prepared as modified Urist's method; the cartilage cells were obtained using mechanical disintegration and enzyme digestion; the BMSCs were separated using whole bone marrow rinsing method, purified, and amplified layer by layer.METHODS: As CFDBM With a composite of different cellular components, and whether applying LIPU stimulation, the samples were divided into four groups: chondrocyte group, BMSCs group, compound group (CFDBM was compounded with chondrocytes,BMSCs, and chondrocytes/BMSCs, respectively, without LIPU stimulation), and LIPU group (CFDBM was compounded with chondrocytes/BMSCs, and then the samples were stimulated with LIPU on the second day, 1.0 MHz frequency, 10 mW/cm~2 transient spatial intension, 20 min/d).MAIN OUTCOME MEASURES: ① the 2~(nd)-generation of cartilage cells and BMSCs were examined by immunohistochemical method; ② The CFDBM prepared as modified Urist's method was examined as HE staining; ③ The samples of four groups were examined by collagen II immunohistochemical staining on the 21~(st) day.RESULTS: ① The collagen II immunohistochemical staining of the second generation of the articular cartilage cells showed that the morphostructure was polygon, star or round, and pseudopodia extended, and the cells were rich in cytoplasm; the cytoplasm was brownish yellow, and the cell nuclear was round. ② The result of immunohistochemical staining of BMSCs showed that,CD34 was negative, CD44 and CD105 were positive. ③ In the center of CFDBM prepared as modified Urist's method, there was no obvious cell-like structure and the gap size was uniform. ④ On the 21~(st) day after combining CFDBM with cells, collagen II immunohistochemical staining demonstrated that BMSCs group was negative, chondrocyte group was weak positive, compoundgroup was positive, and the LIPU group was strongly positive.CONCLUSION: ① Biological property of the 1~(st)-3~(rd)-passage chondrocytes and BMSCs was similar to primary-cultured cells. ②Both chondrocytes and BMSCs had a highly proliferative ability in CFDBM. ③ 10 mW/cm~2 LIPU could not affect activity of BMSCs but could promote differentiation Into articular cartilage cells, and it also could not promote celt proliferation.
6.A study on anti-inflammatory and analgesic effects of alkaloids of Toddalia asiatica
Xiaoyan HAO ; Lin PENG ; Lang YE ; Nenghui HUANG ; Yuemao SHEN
Journal of Integrative Medicine 2004;2(6):450-2
OBJECTIVE: To study the pharmacological activities and toxicity of the crude alkaloids of Toddalia asiatica and to provide pharmacological data for the further development of this herbal medicine. METHODS: We observed the anti-inflammatory effects of the crude alkaloids of Toddalia asiatica, using xylol and agra to induce the turgidness and sodium carboxymethyl cellulose (CMC-Na) to induce leucocyte strolling in the rats. The analgesic effects were observed by body-distortion methods. The effects of alkaloids of Toddalia asiatica on hepatic function were observed by testing the contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum and calculating the liver index. The LD 50 and 95% creditability were calculated with developed Karber Method. RESULTS: The administration of alkaloids of Toddalia asiatica had the function of inhibiting the auricle swelling caused by xylol and joint swelling caused by agar and leucocyte migration caused by CMC-Na, decreasing the body-distortion of the rats. After two weeks administration, the contents of ALT and AST showed that there was no obvious difference between administered group and control group. The LD50 of the crude alkaloids of Toddalia asiatica was 1.622 g/kg and the 95% creditability was 1.29-2.03 g/kg. CONCLUSION: Toddalia asiatica has anti-inflammatory and analgesic effects, and there is no injury to the liver after long-term administration in rats.
7.Clinical significance of urinary mALB ,RBP and CysC and their combined detection in early diagnosis of type 2 diabetic nephropathy*
Shaoxiong ZHOU ; Yongjian SHEN ; Lin ZOU ; Lishan HUANG
International Journal of Laboratory Medicine 2017;38(6):786-787,791
Objective To investigate the application value of urine microalbumin (mALB) ,retinol binding protein(RBP) and cys-tatin C(CysC) and their combined detection in early diagnosis of type 2 diabetic nephropathy(DN) .Methods Ninety-two inpatients with DN (DN group) and 90 people undergoing the physical examination(control group) in our hospital from June 2014 to Decem-ber 2015 were collected .Urine mALB ,RBP and CysC were detected in all subjects and detection results were analyzed statistically . Results The levels of urine mALB ,RBP and CysC in the DN group were significantly higher than those in the control group ,the differences all had statistical significance (P< 0 .05) .Among 3 indicators ,the positive rate of urine mALB for detecting DN was highest (94 .57% ) ,while which of 3-index combined detection was 97 .83% ,and significantly higher than that of single detection , the difference was statistically significant(P<0 .05) .The sensitivity ,specificity ,positive predictive value ,negative predictive value and Youden index of 3-index combined detection were all higher than those of single index .The ROC curve showed that AUC of u-rine mALB for diagnosing DN was 0 .732 ,the diagnostic cut-off value was 43 .58 mg/L ,AUC of urine RBP was 0 .685 ,the diagnos-tic cut-off value was 1 .47 mg/mL ,AUC of urine CysC was 0 .701 ,the diagnostic cut-off value was 1 .42 mg/L ,while AUC of com-bined detection was 0 .928 .Conclusion Urine mALB ,RBP and CysC are better indexes reflecting renal injury .Their combined de-tection will increase the positive rate ,sensitivity and specificity for diagnosing DN .So monitoring the levels of urine mALB ,RBP and CysC has an important significance to diagnosing the occurrence and development of DN early renal injury and prevention ,treat-ment and delaying progress of DN .
8.Effects of carbon monoxide releasing molecule-2 on post-resuscitation myocardial dysfunction
Shen ZHAO ; Yumin HE ; Qingming LIN ; Feng CHEN ; Zitong HUANG
Chinese Journal of Emergency Medicine 2016;25(12):1278-1283
Objective To investigate the protective role of carbon monoxide releasing molecule-2 (CORM-2) in post-resuscitation myocardial dysfunction (PRMD) in rat models of cardiopulmonary resuscitation (CPR).Methods Cardiopulmonary resuscitation model was established after cardiac arrest induced by ventricular fibrillation.Male healthy Sprague-Dawley (SD) rats were randomly (random number) divided into 4 groups according to random number table:control group,CORM-2 group,inactive CORM-2 (iCORM-2) group and Sham group,in which the equal volume (1 mL) of 0.2% DMSO,50 μmol/kg CORM-2,50 μmol/kg iCORM-2 and 0.2% DMSO were respectively administered into the rats of these groups after resuscitation.The ejection fraction (EF) of left ventricle and myocardial performance index (MPI) were measured to detect the myocardial function by echocardiography at 12 hours after resuscitation.Mitochondrial respiration was assessed with Clark oxygen electrode at the same time.Western blot was used to determine the ratio of mitochondrial cytochrome c (cyt c) to cytoplasmic cyt c as well as caspase-3 level.Multiple comparisons were made by analysis of variance.Results Compared with the control group,higher EF and MPI,higher state Ⅲ respiration rate and respiratory control rate (RCR) of mitochondria,and decreased ratio of mitochondrial cytc/cytoplasmic cyt c and lower caspase-3 level were observed in the CORM-2 group (P < 0.05).However,there were no significant differences in above biomarkers found between iCORM-2 group and control group (P > 0.05).Conclusions The CO released from CORM-2 might improve mitochondrial respiration and PRMD by inhibition of myocardial apoptosis via a mitochondrial pathway.
9.Construction of lentiviral vector of RNA interference and its effect on A549 and A549/DDP cells
Jianfeng HUANG ; Zhenya SHEN ; Lin XU ; Yunsheng YU ; Wenxue YE ; Haoyue HUANG
China Oncology 2009;19(7):497-502
Background and purpose: Multidrug resistance (MDR) of tumor cell is the main reason for clinical chemotherapy failure as well as cancer recurrence and metastasis. This study was to construct a lentiveral vector of RNA interference of MDR1 gene and observe its inhibitive role on the expression of MDR1 in A549 and A549/DDP cells. Methods: Oligos of base pairs for hairpin RNA targeting MDR1 were chemically synthesized. Via annealing and inserting them into the down-stream of H1 promoter of linearized pSUPER, we obtained the siRNA constructs for MDR1, which were afterwards transfected into A549, a human lung cancer cell line expressing high level MDR1, the impact of constructs was observed on the expression and interference efficiency of siRNA against MDR1. The effective sequence of siRNA targeting MDR1 gene was confirmed. Both sense and antisence oligo DNA of the targeting sequence was designed, synthesized and cloned into the PTM vector, containing a promoter and a green fluorescent protein (GFP). The resulting lentiviral vector containing MDR1 siRNA was named PTM-siMDR1 and then transfected into A549 and A549/DDP cells after being confirmed by PCR and sequencing. Results: Restriction digestion and DNA sequencing showed that the siRNA constructs for MDR1 were successfully produced and the expressed siRNA could effectively down-regnlate the expression of MDRI. PCR demonstrated that the lentivirus RNAi vector of MDR1 producing PTM-siMDR1 was constructed successfully. The chemosensitivity of A549/DDP cells to cisplatin were enhanced obviously after trartsfection. Conclusion: The lentivirus RNAi vector of MDR1 can significantly revise the resistance ofA549/ DDP cells with eisplatin after infection.
10.HLA-A2-restricted point-mutated Survivin peptide elicited specific cytotoxic T lymphocyte reaction against hepatocarcinoma
Han SHEN ; Zhaoliang HUANG ; Xiaohua CHEN ; Yanmei LIN ; Teng WANG ; Hongwei SHAO ; Shulin HUANG
Chinese Journal of Microbiology and Immunology 2012;32(8):696-701
Objective To investigate the anti-hepatocarcinoma(anti-HCC) function of HLA-A2-restricted point-mutated Survivin peptide induced CTLs.Methods The HLA-A2-restricted Survivin nonapeptides were evaluated using bioinformatics software.The binding affinity of Survivin peptide to HLA-A2 molecular was determined with flow cytometry analysis.After peptide-induced CTLs were generated in vitro,flow cytometry and ELISA were performed to detect the levels of IFN-γ,which were secreted by reactive CTLs.Peptide-induced CTLs were co-cultured with hepatoma cell lines HepG2 and BEL-7402.The rates of tumor cells lysis were assayed using CytoTox 96(R) and the morphological changes of tumor cells were observed with inverted phase contrast microscope.Results Point-mutated Survivin nonapeptide Sur79M2 (KMSSGCAFL) was filtered out,which was shown higher scores compared with the wild-type peptide Sur79.Consistent with the results of software analysis,Sur79M2 showed higher binding ability in T2 binding assays.At the same time,Sur79M2-induced CTLs could release a large number of IFN-γ after incubated with target cells rather than Sur79.When co-cultured with HCC cell lines HepG2 and BEL-7402,Sur79M2-induced CTLs effectively lysis HepG2 on HLA-A2-restricted manner without killing effect on BEL-7402 that do not express HLA-A2 molecules.Conclusion Sur79M2 could elicited specific cytotoxic T lymphocytes in vitro,which were able to specifically kill HCC cell lines on HLA-A2-restricted manner.