1.Clinical significance of inflammatory biomarkers in distinguishing concurrent bacterial infection from idiopathic inflammatory myopathy
Yizhi XIAO ; Hui LUO ; Yaou ZHOU ; Sijia LIU ; Xiaoxia ZUO ; Yisha LI
Chinese Journal of Rheumatology 2017;21(4):258-262
Objective To understand the diagnostic values of procalcitonin (PCT),C-reactive protein (CRP),erythrocyte sedimentation rate (ESR),white blood cell (WBC) and neutmphilic granulocyte ratio (NE%) in distinguishing concurrent bacterial infection from idiopathic inflammatory myopathy (ⅡM).Methods Clinical data and laboratory examinations of 118 ⅡM patients were collected.The ⅡM patients were assigned to the bacterial infection group (n=66) or the non-infection group (n=52).The levels of PCT,CRP,ESR,WBC and NE% were compared by the Mann-Whitney U tests between the two groups and receiver operating characteristic curves were generated in order to evaluate the diagnostic value.Results The levels of PCT (0.06 ng/ml,0.03 ng/ml,U=2.637,P<0.01);CRP (15.80 mg/L,4.40 mg/L,U=5.944,P<0.01);ESR (43.50 mm/1 h,27.00 mm/1 h,U=2.266,P<0.05);WBC (9.85×109/L,7.70×109/L,U=2.675,P<0.01) and NE% (80.70%,75.75%,U=2.344,P<0.01) were significantly higher in the ⅡM patient group with concurrent infection than in the noninfection ⅡM patient group.CRP showed the highest diagnostic value with sensitivity,specificity,positive predictive value and negative predictive value of 72.7%,82.7%,84.2% and 70.5%,respectively.Conclusion The inflammatory biomarkers PCT,CRP,ESR,WBC and NE% offer diagnostic accuracy in detecting bacterial infection in ⅡM patients.Particularly,CRP is the most sensitive and specific biomarker indetecting bacterial infection in ⅡM patients.
2.Effects of α-enolase silencing on drug resistance in drug resistant cell line K562/A02
Xue GAO ; Zhou YE ; Kexiong WU ; Dongmei FAN ; Ming YANG ; Yanjun ZHANG ; Yizhi ZHANG
Chinese Pharmacological Bulletin 2014;(11):1521-1525,1526
Aim Drug resistance is one of the major hinders on cancer treatments. α-enolase ( eno1 ) was closely related to the generation and development of drug resistance. This article aims to study the effect of eno1 on cell growth and drug resistance in human chro-nic myeloid leukemia cell line K562/A02 . Methods We screened three eno1 stable silencing cells K562/A02-sheno1 and its control cells K562/A02-shcon. Cell count assay was performed to test cell growth, MTT assay was used to test cell proliferation, flow cytometry was used to test the intra-cellular Rho123 content, the expression of genes were tested by real-time PCR assay and western blot assay on mRNA level and protein level, respectively. Results eno1 was o-ver-expressed in K562/A02 cells and its expression was increased by ( 2. 85 ± 0. 56 ) times and ( 1. 43 ± 0. 05 ) times on mRNA level and protein level com-pared to K562 cells. However, there was no difference in cell growth rate between K562/A02 cells and K562 cells. K562/A02-sheno1 cells showed lower cell growth rate and higher drug sensitivity to anti-cancer drugs taxol and doxorubicin. Moreover the Rho123 content was increased in K562/A02-sheno1 cells. The expression of MDR1 decreased in both mRNA level and protein level in K562/A02-sheno1 cells. Conclusion eno1 silencing could suppress cell growth, reverse drug resistance and increase its drug sensitivity in K562/A02 cells, and the mechanism was associated with the MDR1 gene.
3.Location and relative quantity of flavonoids in the leaf of Apocynum venetum.
Hu XU ; Mei WANG ; Xunhong LIU ; Jianya XU ; Xingsheng FU ; Yizhi ZHOU
Acta Pharmaceutica Sinica 2011;46(8):1004-7
In this study, laser scanning confocal microscopy (LSCM) was used to determine the location and relative quantity of flavonoids in the leaves of Apocynum venetum L. from the top, middle and basal parts of the branch. The leaves of the plants of one, two and three years old, separately, were collected in July. ANOVA and LSD test were employed in the statistical analysis. The results indicated that flavonoids located mainly in xylem conduit of vein, collenchyma, epidermic cells and cuticle. The data of flavonoids contents under statistical analysis showed that difference existed in the leaves of different parts and different ages. This study provided the reliable scientific material about the analysis of the ecological and the exploitation of the leaves of Apocynum venetom L.
4.Lentivirus-mediated inhibition of tumor necrosis factor-alpha effect in expression of Caspase-3 and Bcl-2 in spinal cord contusion rats
Yizhi WANG ; Xi HU ; Xiuya ZHOU ; Hongyu XIANG ; Ya LIN ; Bingyi TANG ; Tingting TANG ; Qian ZENG ; Xiao ZHANG
The Journal of Practical Medicine 2016;32(17):2807-2812
Objective To explore the change of apoptosis factor Caspase-3 and Bcl-2 in the injured segment of rat with spinal cord injury after inhibiting lentivirus expression of inflammation factor TNF-α. To study the relationship between Caspase-3, Bcl-2, Bax and TNF-α in spinal cord injury. Mthods Spinal cord contusion model was prepared by Allen method. The relation between tumor necrosis factor alpha and Bcl-2, was predicted by the method of GeneMANIA bioinformatics. The RNA which was packaged by lentivirus constructed the RNA interference model of tumour necrosis factor alpha. After interference of tumor necrosis factor alpha, we used the method of QRT-PCR to assays the mRNA expression of Caspase-3 and Bcl-2 in spinal cord and detect of the localization of Caspase-3 and Bcl-2 by immunohistochemistry. Statistical analysis with SPSS17.0. Results SD rats had paraplegia and urinate retentaion because of spinal cord injury. The result of QRT-PCR showed that in the seventh day after SCC, the expression of Caspase-3 reduced significantly (P < 0.05) and Bcl-2 did not change significantly (P > 0.05). Immunohistochemistry experiment results showed that Caspase-3 Bcl-2 and Bax immunoreactive cells were observed in the neurons and glial cells of both white matter and gray matter in the spinal cord. The results were the same with QRT-PCR.. Conclusion TNF-α in rats after SCC can effectively regulate the ratio of Bcl-2 and Bax , and then regulate the expression of Caspase-3 , which may affect the function of apoptosis and function recovery after spinal cord injury.
5.Study on water-soluble iron, heavy metals and harmful elements of Magnetitum.
Xingsheng FU ; Xunhong LIU ; Ruichao LIN ; Hu XU ; Yizhi ZHOU ; Dekang WU ; Shengjin LIU
China Journal of Chinese Materia Medica 2011;36(12):1572-1576
OBJECTIVETo measure the contents of the water-soluble iron, five heavy metals and harmful elements in Magnetiturn and provide a basis for the quality control and safety evaluation of Magnetitum.
METHODIron (Fe), lead (Pb), cadmium (Cd) and copper (Cu) were determined by atomic absorption spectrometry (AAS); arsenic (As) and mercury (Hg) were determined by atomic fluorescence spectrometry (AFS).
RESULTThe mean content of element iron is 764.30 mg x kg(-1). The contents of five water-soluble heavy metals and harmful elements in Magnetitum were within the safety range. The recovery of the standard addition was in the range of 93.7% - 110.6%, and the RSD was less than 5.0%.
CONCLUSIONAnalyzing the water-soluble iron, heavy metals and harmful elements in Magnetitum is effective to the quality control and the safety evaluation of magnetitum.
Iron ; metabolism ; Materia Medica ; chemistry ; Metals, Heavy ; metabolism ; Solubility ; Spectrophotometry, Atomic
6. In vitro antibacterial activity of triclosan in combination with different antibacterial agents against triclosan-resistant multidrug-resistant Acinetobacter baumannii
Ye XU ; Yizhi ZHANG ; Chunquan XU ; Siqin ZHANG ; Xiucai ZHANG ; Wenya XU ; Tieli ZHOU
Chinese Journal of Microbiology and Immunology 2019;39(9):674-679
Objective:
To investigate the
7.Resistance mechanisms and molecular epidemiology of chlorhexidine acetate-resistant Klebsiella pneu-moniae clinical isolates
Yizhi ZHANG ; Xiucai ZHANG ; Siqin ZHANG ; Yajie ZHAO ; Guofeng DONG ; Xiaozhen ZHOU ; Tieli ZHOU
Chinese Journal of Microbiology and Immunology 2019;39(3):202-207
Objective To investigate the chlorhexidine acetate-resistance in Klebsiella pneumoniae ( K. pneumoniae) clinical isolates and to analyze the possible mechanisms and molecular epidemiology of re-sistant isolates. Methods A total of 332 K. pneumoniae clinical isolates were collected in the First Affilia-ted Hospital of Wenzhou Medical University in 2015. Standard agar dilution was used to screen chlorhexidine acetate-resistant isolates. The minimum inhibition concentrations ( MIC) of chlorhexidine acetate to resistant isolates with and without the presence of carbonyl cyanide m-chlorophenyl hydrazone ( CCCP) , which was an efflux pump inhibitor, were analyzed. Efflux pump genes of cepA, qacE and qacΔE1 that carried by and ex-pressed in those isolates were detected by polymerase chain reaction ( PCR) and quantitative real-time PCR ( RT-qPCR) , respectively. The biofilm formation ability was measured by crystal violet staining. The homol-ogy among the chlorhexidine acetate-resistant isolates was investigated with multilocus sequence typing ( MLST) and pulsed-field gel electrophoresis ( PFGE) . Results Twenty-five K. pneumoniae strains were re-sistant to chlorhexidine acetate. The MIC values of chlorhexidine acetate for them were reduced by at least four-fold in the presence of CCCP. Strains carrying the genes of cepA, qacE and qacΔE1 accounted for 100%, 40% and 40%, respectively. The expression of the efflux pump genes in the chlorhexidine acetate-resistant isolates was higher than that in the susceptible isolates. The biofilm formation ability of the chlo-rhexidine acetate-resistant isolates was better than that of the susceptible isolates. Furthermore, negative, weak-positive and positive biofilm formation ability was observed in four ( 16%) , 20 ( 80%) and one (4%) strains, respectively. The results of MLST and PFGE showed that the 25 chlorhexidine acetate-resist-ant isolates belonged to 19 different sequence types ( ST) with diverse PFGE patterns. Conclusions This study suggested that active efflux was the main mechanism of chlorhexidine acetate resistance in K. pneumoni-ae. The 25 chlorhexidine acetate-resistant K. pneumoniae strains possessed different biofilm formation ability and shared low homology.
8.Effects of estrogen on epidermis growth of mice and proliferation of human epidermal cell line HaCaT and its mechanism.
Tao ZHOU ; Jing CHEN ; Zongwei HUANG ; Li FANG ; Yu CHEN ; Yajie CHEN ; Yizhi PENG
Chinese Journal of Burns 2016;32(5):299-304
OBJECTIVETo observe the effects of estrogen on epidermis growth of mice and proliferation of keratinocytes (human epidermal cell line HaCaT), and to explore its mechanism.
METHODS(1) Five adult C57BL/6 mice in estrus cycle were identified by vaginal exfoliative cytology diagnosis and set as estrus group, while another 5 adult C57BL/6 mice with ovary resected before sexual development were set as ovariectomized group. The full-thickness skin from the tail root of mice in two groups were collected. The thickness of epidermis was observed and measured after HE staining. The distribution of proliferating cell nuclear antigen (PCNA)-positive cells in epidermis was observed by immunohistochemical staining, the number of which was counted. (2) HaCaT cells in logarithmic growth phase were cultured with RPMI 1640 nutrient solution containing 10% fetal bovine serum, and they were divided into negative control group (NC), pure estradiol group (PE), protein kinase B (Akt) inhibitor group (AI), and extracellular signal-regulated kinase (ERK) inhibitor group (EI) according to the random number table, with 20 wells in each group. To nutrient solution of each group, 1 μL dimethyl sulfoxide, 1 μL 17β-estradiol (100 nmol/L), 1 μL LY294002 (10 μmol/L), and 1 μL PD98059 (30 μmol/L) were added in group NC, group PE, group AI, and group EI respectively, and the last two groups were added with 1 μL 17β-estradiol (100 nmol/L) in addition. At post culture hour (PCH) 0 (immediately after culture), 24, 48, 72, 5 wells of cells from each group were collected to detect the proliferation activity of cells by cell counting kit 8 and microplate reader. (3) HaCaT cells in logarithmic growth phase were collected, grouped, and treated with the above-mentioned methods, with 3 wells in each group. At PCH 72, cell cycle distribution was detected by flow cytometer to calculate proliferation index (PI) of cells. (4) HaCaT cells in logarithmic growth phase were collected, grouped, and treated with the above-mentioned methods, with 3 dishes in each group. At PCH 72, the protein levels of phosphorylated Akt (p-Akt), phosphorylated ERK (p-ERK), and PCNA were determined with Western blotting. The cell experiments were repeated for 3 times. Data were processed with t test, one-way analysis of variance, analysis of variance of factorial design, and LSD test.
RESULTS(1) The epidermis thickness of mice in ovariectomized group was (33.5±3.0) μm, which was obviously thinner than that in estrus group [(51.4±3.1) μm, t=20.7, P<0.01]. The PCNA-positive cells mainly aggregated in the basal layer of epidermis of mice in two groups. The number of PCNA-positive cells in epidermis of mice in ovariectomized group was 37±12 per 200 fold visual field, obviously fewer than that in estrus group (96±15 per 200 fold visual field, t=15.3, P<0.01). (2) During PCH 0 to 48, there were no significant differences in the proliferation activity of cells between group PE and group NC (with P values above 0.05). At PCH 72, compared with that in group NC, the proliferation activity of cells in group PE was obviously increased (P<0.01). The proliferation activity of cells in groups AI and EI was obviously lower than that in the previous two groups (with P values below 0.01). (3) Compared with that in group NC [(51.6±1.1)%], the PI of cells in group PE was obviously increased [(58.5±0.8)%, P<0.05]. The PI values of cells in groups AI and EI were (34.9±0.8)% and (48.2±0.4)% respectively, both obviously lower than those in the previous two groups (with P values below 0.01). (4) Compared with that of group NC (0.566±0.034), the protein level of p-Akt in cells of group PE was significantly increased (1.048±0.077, P<0.01). Compared with that of group PE, the protein level of p-Akt was obviously decreased in cells of groups AI and EI (respectively 0.682±0.095 and 0.672±0.019, with P values below 0.01). Compared with that of group NC (0.469±0.013), the protein level of p-ERK obviously increased in cells of groups PE, AI, and EI (respectively 1.064±0.089, 1.010±0.038, 0.778±0.065, with P values below 0.01). The protein level of p-ERK in cells of group EI was obviously lower than that in group PE (P<0.01). Compared with that of group NC (0.386±0.053), the protein level of PCNA was obviously increased in cells of group PE (0.743±0.043, P<0.01). The protein levels of PCNA in cells of groups AI and EI were 0.264±0.019 and 0.223±0.065 respectively, both obviously lower than those in the previous two groups (with P values below 0.01).
CONCLUSIONSLack of estrogen damages the growth ability of epidermis of mice. Estrogen (17β-estradiol) can promote the proliferation of HaCaT cells by increasing the expression of PCNA via activating ERK/Akt signaling pathway.
Animals ; Cell Cycle ; Cell Line ; Cell Proliferation ; drug effects ; Epidermis ; cytology ; drug effects ; growth & development ; Estradiol ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Female ; Humans ; Keratinocytes ; cytology ; drug effects ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Proliferating Cell Nuclear Antigen ; metabolism ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; Signal Transduction
9.Analysis on the prevalence of central venous catheter-related infection in burn patients and its risk factors.
Li FANG ; Fan WANG ; Kedai SUN ; Tao ZHOU ; Yali GONG ; Yizhi PENG
Chinese Journal of Burns 2016;32(4):243-248
OBJECTIVETo investigate the prevalence of central venous catheter-related infection (CRI) in burn patients and its risk factors, so as to guide the clinical practice.
METHODSClinical data of 5 026 days of 480 cases of central venous catheterization altogether in 228 burn patients admitted to our ward from June 2011 to December 2014, conforming to the study criteria, were retrospectively analyzed. (1) The incidence of CRI and that of catheter-related bloodstream infection (CRBSI) in patients (the infection rates per thousand days were calculated) and mortality due to them, and detection of concerning bacteria were recorded after each case of catheterization. (2) The incidence of CRI after each case of catheterization in patients was recorded according to the classification of their gender, age, total burn area, full-thickness burn area, cause of injury, severity of inhalation injury, location of catheterization, whether catheterization through wound or not, duration of catheterization, and the data were processed with chi-square test. Indexes with statistically significant differences were selected, and they were processed with multivariate logistic stepwise regression analysis to screen the independent risk factors of CRI. (3) To all cases of catheterization and cases with catheterization through wound, incidence of CRI after each case of catheterization in patients at each time period was recorded according to the sorting of duration of catheterization. Data were processed with chi-square test and Fisher's exact test, and the values of P were adjusted by Bonferroni.
RESULTS(1) Infection rate of CRI per thousand days was 50.14‰ (252/5 026), resulting in the mortality rate of 3.51% (8/228). Infection rate of CRBSI per thousand days was 18.70‰ (94/5 026), resulting in the mortality rate of 2.19% (5/228). Respectively 319 and 105 strains of pathogens were detected in CRI and CRBSI, in which the top four bacteria detected were Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae, and the most common fungus found was smooth Candida. (2) There were no statistically significant differences in the incidence of CRI after each case of catheterization among patients with different gender, age, cause of injury, severity of inhalation injury, and location of catheterization (with χ(2) values from 0.427 to 6.991, P values above 0.05). There were statistically significant differences in the incidence of CRI after each case of catheterization among patients with different total burn area, full-thickness burn area, whether catheterization through wound or not, duration of catheterization (with χ(2) values from 7.202 to 14.246, P<0.05 or P<0.01). (3) Total burn area, whether catheterization through wound or not, and duration of catheterization were the independent risk factors of CRI (with odd ratios respectively 1.495, 1.670, 1.924, 95% confidence intervals respectively 1.096-2.040, 1.077-2.590, 1.303-2.841, P<0.05 or P<0.01). (4) In all cases enduring catheterization, the incidence of CRI in patients after each episode of catheterization was close between cases enduring catheterization shorter than or equal to 3 days and those longer than 3 days and shorter than or equal to 5 days (χ(2) <0.001, P>0.05); the incidence of CRI in patients after each episode of catheterization was significantly higher in cases enduring catheterization longer than 5 days and shorter than or equal to 7 days, longer than 7 days and shorter than or equal to 14 days, and longer than 14 days than the former two periods (with χ(2) values from 3.625 to 13.495, P values below 0.05). In the cases with catheterization through wound, the incidence of CRI of patients after each episode of catheterization was close between cases enduring catheterization shorter than 5 days and those longer than or equal to 5 days and shorter than 7 days (P>0.05); the incidence of CRI of patients after each episode of catheterization was significantly higher in cases enduring catheterization longer than or equal to 7 days and shorter than 14 days and longer than or equal to 14 days than those with longer than or equal to 5 days and shorter than 7 days (with χ(2) values respectively 6.828 and 4.940, P values below 0.05).
CONCLUSIONSThe infection rate of CRI per thousand days in burn patients is relatively low, while that of CRBSI is relatively high, both resulting in relatively low mortality, and Acinetobacter baumannii is the main pathogen. Total burn area, whether catheterization through wound or not, and duration of catheterization are independent risk factors of CRI in burn patients, and with which its occurrence could be predicted. It is suggested that central venous catheterization should be removed within 5 days, and catheterization through wounds should be avoided as much as possible. If catheterization through wound is unavoidable, removal of the catheter within 7 days is recommended.
Acinetobacter baumannii ; isolation & purification ; Burns ; complications ; Catheter-Related Infections ; epidemiology ; Humans ; Incidence ; Prevalence ; Retrospective Studies ; Risk Factors
10.Role of type Ⅵ secretion system in the pathogenicity and antibiotic resistance of Acinetobacter baumanii
Kaihang YU ; Lijiang CHEN ; Renchi FANG ; Wenzi BI ; Xiucai ZHANG ; Yizhi ZHANG ; Tieli ZHOU ; Jianming CAO
Chinese Journal of Infectious Diseases 2020;38(4):231-236
Objective:To investigate the role of type Ⅵ secretion system (T6SS) in the pathogenicity and antibiotic resistance of Acinetobacter baumanii. Methods:From January 1 to December 31, 2016, a total of 45 Acinetobacter baumanii isolates were collected from patients with bloodstream infection in the First Affiliated Hospital of Wenzhou Medical University. The susceptibilities to commonly used antimicrobial agents were determined by VITEK 2 Compact automatic microbiology analyzer. Detection of T6SS characteristic gene hemolysin coregulated protein ( hcp) was achieved by polymerase chain reaction. Biofilm formations, serum resistances and competition tests of T6SS-positive/negative Acinetobacter baumanii were performed in vitro. The clinical data of patients with bloodstream infection were collected and analyzed. Chi-square test, t test and Kruskal-Wallis test were conducted for statistical analysis. Results:The positive rate of T6SS in 45 Acinetobacter baumanii isolates was 53.3% (24/45). The resistance rates of T6SS-positive Acinetobacter baumanii to ceftazidime, ciprofloxdcin, gentamicin, imipenem, levofloxacin, piperacillin/tazobactam, tobramycin and cefepime (95.8%, 95.8%, 66.7%, 95.8%, 79.2%, 95.8%, 79.2%, 91.7%)were all higher than that of T6SS-negative Acinetobacter baumanii (28.6%, 28.6%, 28.6%, 28.6%, 9.5%, 23.8%, 23.8%, 28.6%), and the differences were all statistically significant ( χ2=22.12, 22.12, 6.51, 22.12, 21.83, 24.72, 13.79, 18.97, respectively, all P<0.05). The biofilm formation ability, serum resistance and competitive ability of T6SS-positive Acinetobacter baumanii were stronger than those of T6SS-negative Acinetobacter baumanii, and the differences were all statistically significant ( t=4.99, Z=-2.61 and -2.27, respectively, all P<0.05). The positive rate of T6SS isolated from intensive care unit (ICU) ward (80.0%, 16/20) was significantly higher than that from non-ICU ward (32.0%, 8/25; χ2=10.29, P<0.05). But T6SS had no effect on the prognosis of patients ( χ2=1.74, P=0.188). Conclusions:T6SS of Acinetobacter baumanii is associated with high pathogenicity, and the high drug resistance rate makes treatment extremely difficult. Physicians need to pay much attention, especially to the patients from ICU wards.