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Objective To investigate the expression differences and significance of periostin (PN) in eyelid basal cell carcinoma associated fibroblasts (BCAFs) andnormal fibroblasts (NFs) after separation,culture,purification and identification.Methods The third generation of purified BCAFs and NFs was selected,and the concentrations of cell suspensions were modulated to 20 × 106 L-1 by trypsin,and then the cell suspension were seeded and cultured in 6-well plate by 2 mL per well.The cell culture supernatants were collected when BCAFs and NFs were cultured by serum-free medium for 48 h,then the content of PN in cell culture supernatants from BCAFs and NFs was detected by enzyme-linked immunosorbent assay (ELISA).The glass coverslips were placed at the bottom of the 6-well plate to make cell slides,and then the expression of PN in BCAFs and NFs cells were tested by immunofluorescence staining.Results ELISA showed that the content of PN in cell culture supernatants from BCAFs and NFs was (9.26 ± 2.35) μg · L-1 and (2.57 ± 0.41) μg · L-1.And the expression level of PN in BCAFs tested by immunofluorescence staining technology was higher than that in NFs cells,and the differences were statistically significant (all P ＜ 0.05).Conclusion The expression and secretion of PN in the eyelid BCAFs were highly enhanced when compared with NFs,suggesting that periostein may promote or inhibit the occurrence and development of the eyelid basal cell carcinoma in the microenvironment of the eyelid basal cell carcinoma.
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Several studies have demonstrated an association between physical activity and telomere length; however, the association remains inconsistent. A cross-sectional study consisting of 588 participants (375 females, median age of 33.8 years) was carried out to investigate the association between telomere length and physical activity in a general population from North China. The results show that relative telomere length is not significantly different in participants in the northern Chinese population with different levels of physical activity, either in the model only adjusted for age (F = 2.127, P = 0.120) or in the model adjusted for demographics and lifestyle (F = 1.227, P = 0.294). The gender-stratified analysis also produced insignificant results. Our study confirmed a non-significant association between physical activity and telomere length in the northern Chinese population, which adds to the inconsistent association between physical activity and telomere length across different ethnic populations.
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Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Povo Asiático , Genética , China , Estudos de Coortes , Estudos Transversais , Exercício Físico , Fisiologia , TelômeroRESUMO
<p><b>OBJECTIVE</b>To observe the effects of Xiongshao Capsule (, XSC) on anti-inflflammatory properties of high-density lipoprotein (HDL), myeloperoxidase (MPO) and paraoxonase 1 (PON1) in serum of atherosclerosis (AS) rabbit model and explore the anti-inflflammatory protective effects of XSC on HDL.</p><p><b>METHODS</b>Sixty rabbits were randomized into the control, the model, XSC low-, medium- and high-dose (Rhizoma Chuanxiong + Radix Paeoniae rubra: 0.6+0.3, 1.2+0.6, 2.4+1.2g·kg·day, respectively), and simvastatin (1g·kg·day) groups. The model rabbits were fed with high-fat diet and drugs for 15 weeks. The blood and thoracic aortas samples were collected at the end of 15 weeks. The levels of serum MPO and PON1 as well as total cholesterol (TC) and free cholesterol (FC) in aorta wall cells were tested by enzyme linked immunosorbent assay.</p><p><b>RESULTS</b>TC and FC in the model group were significantly higher than those in the control group (P<0.01). Compared with the model group, TC and FC in the XSC groups were signifificantly lower (P<0.05 or P<0.01), so was simvastatin group (P<0.01). There was no signifificant difference in PON1 level between groups (P>0.05), even between model and control groups (P>0.05). The serum MPO level in the model group was signifificantly higher than that in the control group (P<0.05), which was signifificantly lower in XSC groups as well as simvastatin group (P<0.05 or P<0.01), and no difference was found between XSC groups and simvastatin group (P>0.05).</p><p><b>CONCLUSIONS</b>XSC can reduce the serum MPO level in AS rabbits to protect the anti-inflammatory function of HDL, maintaining the normal lipid transport function. TC and FC levels in aorta cells decline, and this process initiated by XSC plays an anti-AS role.</p>
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Interventional stents are emerging medical apparatus and instruments which can be implanted into the lesion location to reopen and scaffold the blocked cavities. The ideal stents must have reliable mechanical properties, and the current research methods on mechanical properties of the stents are mainly based on theoretical analysis, numerical analysis and experimental test. In this article, all kinds of analytic methods on testing mechanical properties of the stents and their limitations are discussed and summarized in detail, and further research on the stents is prospected as well.
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BACKGROUND:Parathyroid hormone can activate a series of physiological and biochemical reactions through specific receptors on the surface of target organs,to adjust the dynamic calcium balance.OBJECTIVE:To explore the mechanisms whereby parathyroid hormone maintains in vitro proliferation of bone marrow mesenchymal stem cells (BMSCs),by observing the effects of parathyroid hormone on calcium ions in BMSCs.METHODS:BMSCs at passage 4 were randomly divided into two groups.Experimental group was treated with parathyroid hormone,and control group was treated with routine culture medium.During 10 days of culture,MTT assay was used to detect BMSCs proliferation.At 7 and 14 days after treatment,laser scanning confocal microscopy was used to detect fluorescence intensity of calcium ions in BMSCs in the two groups.RESULTS AND CONCLUSION:(1) At 1 and 2 days after culture,the cell viability in the two groups was sluggish.At 3 days after culture,the cells turned into the rapid growth.At 7 and 8 days after culture,the cell growth turned into the plateau.At 10 days after culture,the cell proliferation ability had a weakening trend,which was lower in the control group than in the experimental group (P ＜ 0.05).(2) At 7 days after culture,the fluorescence intensity of intracellular calcium ions was strong.As the induction time extended,the fluorescence intensity of intracellular calcium ions was declined at 14 days after-culture.At 7 and 14 days after culture,the fluorescence intensity of intracellular calcium ions in the experimental group was higher than that in the control group (P ＜ 0.001).There was no difference in the fluorescence intensity of intracellular calcium ions between experimental group at 14 days and control group at 7 days.To conclude,parathyroid hormone could increase the fluorescence intensity of calcium ions in BMSCs,and function to maintain the BMSCs proliferation.
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Objective To analyse the clinical effect of Quantianma capsules combined with flupentixol and melitracen tablets on chronic migraine. Methods 130 patients with chronic migraine were treated with flupentixol and melitracen tablets, were divided into control group and observation group, 65 patients in each group. The observation group were given to Quantianma capsules. Results The effective rate of observation group with 90.77% was higher than control group with 73.85%(P<0.05). After treatment, the number of migraine attacks, duration time, VAS score, HAMA and HAMD score and mean flow velocity of cerebral artery of two groups were lower than pre-treatment and observation group were lower than that of control group(P<0.05). Adverse reactions were similar in the two groups. Conclusion Quantianma capsules can improve the efficacy of treatment of chronic migraine combined with flupentixol and melitracen tablets and improve condition with few adverse effects.
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Objective To analyse the clinical effect of Quantianma capsules combined with flupentixol and melitracen tablets on chronic migraine. Methods 130 patients with chronic migraine were treated with flupentixol and melitracen tablets, were divided into control group and observation group, 65 patients in each group. The observation group were given to Quantianma capsules. Results The effective rate of observation group with 90.77% was higher than control group with 73.85%(P<0.05). After treatment, the number of migraine attacks, duration time, VAS score, HAMA and HAMD score and mean flow velocity of cerebral artery of two groups were lower than pre-treatment and observation group were lower than that of control group(P<0.05). Adverse reactions were similar in the two groups. Conclusion Quantianma capsules can improve the efficacy of treatment of chronic migraine combined with flupentixol and melitracen tablets and improve condition with few adverse effects.
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Interventional stents are emerging medical apparatus and instruments which can be implanted into the lesion location to reopen and scaffold the blocked cavities.The ideal stents must have reliable mechanical properties,and the current research methods on mechanical properties of the stents are mainly based on theoreti cal analysis,numerical analysis and experimental test.In this article,all kinds of analytic methods on testing mechanical properties of the stents and their limitations are discussed and summarized in detail,and further research on the stents is prospected as well.
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<p><b>OBJECTIVE</b>To evaluate the efficacy of internal application of Qigui Mixture (QM) and external application of Qigui Huoxue Lotion (QHL) in treating type 2 diabetic peripheral neuropathy (DNP) patients of qi-yin deficiency complicated phlegm-dampness blocking collaterals syndrome (QYD-PDBCS), and to primarily discuss its mechanism.</p><p><b>METHODS</b>Totally 62 DPN patients of QYD-PDBCS were randomly assigned to the treatment group (31 cases) and the control group (31 cases). All patients received routine comprehensive therapy. Patients in the control group took Mecobalamine Tablet, 500 microg each time, 3 times per day. Patients in the treatment group additionally took QM, 200 mL per day, twice daily. Besides, they had foot bath in QHL 10 - 15 min every evening for 3 months. The efficacy was assessed by Chinese medical symptom integrals and Toronto clinical scoring system (TCSS) before treatment, 2 and 3 months after treatment. The nerve conduction velocity was determined; the serum levels of total antioxidant capacity (T- AOC), malondialdehyde (MDA), and superoxide dismutase (SOD) were detected 2 and 3 months after treatment.</p><p><b>RESULTS</b>The total effective rates of Chinese medical symptom integrals and TCSS score were obviously higher in the treatment group than in the control group (P < 0.05). The nerve conduction velocity was significantly improved in the treatment group, when compared with before treatment (P < 0.01). There was statistical difference in the nerve conduction velocity difference of right median nerve motor branch, bilateral tibial nerve motor branches, bilateral common peroneal nerve motor branches, bilateral ulnar nerve sensory branches, and left tibial nerve sensory branch (P < 0.05). Compared with before treatment, serum levels of T-AOC and SOD significantly increased, and the level of MDA decreased significantly in the treatment group after 2 and 3 months of treatment (P < 0.01). But only the SOD level increased significantly in the control group (P < 0.01). There was no statistical difference in increased T-AOC level between the two groups after 2 months of treatment (P > 0.05), but there was statistical difference in increased SOD level and decreased MDA level (P < 0.05). There was statistical difference in increased T-AOC and SOD levels and decreased MDA level between the two groups after 3 months of treatment (P < 0.05). No adverse reaction occurred during the therapeutic course.</p><p><b>CONCLUSIONS</b>The internal application of QM and external application of QHL combined with Mecobalamine in treating DPN was safe and effective, with more significant efficacy than using Mecobalamine alone. Its mechanism might be associated with resistance to oxidative stress.</p>
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Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Diabetes Mellitus Tipo 2 , Tratamento Farmacológico , Neuropatias Diabéticas , Tratamento Farmacológico , Medicamentos de Ervas Chinesas , Usos Terapêuticos , Qi , Resultado do Tratamento , Vitamina B 12 , Usos Terapêuticos , Deficiência da Energia Yin , Tratamento FarmacológicoRESUMO
When looking back to the literature regarding insomnia, it is believed that "disharmony of stomach leads to insomnia" is an important mechanism for the attack of insomnia. This theory is widely accepted in clinical practices by ancient and modern acupuncturists, thus the method treating insomnia from the stomach is developed. But deeper research on its mechanism is still lacking. Through correlation between stomach meridian and heart meridian, spleen and stomach being the pivot of rise and decline of qi movement and yin-yang and being the pivot of five zang-viscera housing spirit, modern abdomen-brain theory and experimental research, the mechanism of treating insomnia from the stomach is discussed in this paper, so as to be better to guide clinical acupoint-selection and treatment.
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Humanos , Terapia por Acupuntura , História , China , História Antiga , Medicina na Literatura , Distúrbios do Início e da Manutenção do Sono , Terapêutica , EstômagoRESUMO
The gene expressions of codeinone reductase (COR) and berberine bridge enzyme (BBE) in Papaver somniferum were blocked by RNA hairpin of RNA interference (RNAi). The complete sequences of COR and BBE genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR), the results of homology comparison revealed that the cloned COR and BBE genes had high homology with the other gene family members reported in the GenBank. The target sequences of COR and BBE genes were screened in accordance with the design principle of RNAi, a 643 bp fusion gene was obtained by the method of overlapping PCR, then plant expression vector ihpRNA was constructed based on intermediate vector pHANNIBAL and plant expression vector pCEPSPS. With that 78 transgenic plants were obtained through Agrobacterium-mediated and 17 positive plants were screened by PCR, that could initially indicate that the target fragments of COR and BBE gene had been integrated into tobacco genome.
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Fusão Gênica Artificial , Vetores Genéticos , Álcool Oxidorredutases Dependentes de NAD(+) e NADP(+) , Genética , Oxirredutases N-Desmetilantes , Genética , Papaver , Genética , Plantas Geneticamente Modificadas , Genética , Interferência de RNA , RNA Interferente Pequeno , Nicotiana , Genética , Transformação GenéticaRESUMO
This paper introduces recent advances in research and clinical trials on treating exercise-induced fatigue with acupuncture, focusing on the areas of oxygen free radical scavenging and anti-lipid peroxidation activity, the regulation of immune function, prevention of dysfunction exercise induced hypothalamus-pituitary-gonadal axis, improvement of energy metabolism, the ability to maintain internal environment, the ability to alleviate delayed-on set muscle soreness (DOMS), and anti-central nervous system fatigue. This paper is intended to explain that because of its safety, effectiveness, less adverse reactions, and not involving stimulants, acupuncture has great potential to protect against exercise-induced fatigue. However, there are problems of acupoints selection, manipulation and indicators. Therefore, it needs further investigations to explore the advantages of acupuncture treatment for the exercise-induced fatigue.
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Animais , Humanos , Terapia por Acupuntura , Exercício Físico , Fisiologia , Fadiga , Alergia e Imunologia , Metabolismo , Terapêutica , MoxibustãoRESUMO
Clinical trials are often designed as either pragmatic or explanatory. The pragmatic clinical trials are generally used for measuring the effectiveness of a treatment in common clinical practice, while the explanatory trial for measuring the efficacy of a treatment under ideal conditions. Since the methods concerning pragmatic clinical trials are less introduced in China, its archetypal features, advantages and limitations were introduced in this paper. And a current study of pragmatic clinical trials on using acpuncture for treatment of low back pain carried out in German was taken as an example to illustrate the practical methods concretely. The key steps of the design were presented in detail, and reasonable suggestions about the problems often encountered in the trial, as well as how to balance the internal and external validity, outcome measurement, etc., were offered.
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Humanos , Ensaios Clínicos como Assunto , Métodos , Padrões de Referência , Interpretação Estatística de Dados , Tomada de Decisões GerenciaisRESUMO
<p><b>OBJECTIVE</b>To investigate the mechanism of DDP-resistance in gastric cancer cell line SGC7901/DDP mediated by phosphate(p)-JNK.</p><p><b>METHODS</b>The p-JNK expression was blocked by the JNK inhibitor SP600125. The drug sensitivity was detected by MTT. Cell apoptosis rate was measured by flow cytometry. The expression of p-JNK and P-glycoprotein (P-gp) was examined by Western blot. The expression of both proteins were detected in a tissue microarray containing 168 spots of cancer tissue and 27 spots of normal gastric tissue by SP immunohistochemistry. Pearson method was used to analyze the correlation between p-JNK and P-gp.</p><p><b>RESULTS</b>The drug sensitivity and cell apoptosis rate significantly increased (P<0.01), and the protein expression levels of p-JNK and P-glycoprotein were down-regulated after the inhibition of p-JNK by SP600125 in both SGC7901(p-JNK: 1.17+/-0.03 vs 0.38+/-0.071, P-gp: 0.21+/-0.01 vs 0.06+/-0.01) and SGC7901/DDP (p-JNK: 2.56+/-0.14 vs 1.02+/-0.12, P-gp: 0.77+/-0.05 vs 0.52+/-0.06 )cells(all P<0.01). The protein expression rates of p-JNK and P-glycoprotein were 45.8% and 51.8% respectively in gastric cancer tissue, which were significantly higher than those in normal gastric tissue 7.4% and 18.5% (P<0.01). The correlation of protein expression of p-JNK and P-gp was positive (P<0.01).</p><p><b>CONCLUSION</b>JNK anti-apoptosis pathway with the regulation of P-gp expression plays an important role in the DDP-resistance of gastric cancer, which may be a novel target for reversing multidrug resistance.</p>
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Humanos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP , Metabolismo , Apoptose , Linhagem Celular Tumoral , Cisplatino , Metabolismo , Farmacologia , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Regulação Neoplásica da Expressão Gênica , Proteínas Quinases JNK Ativadas por Mitógeno , Metabolismo , Transdução de Sinais , Neoplasias Gástricas , Tratamento Farmacológico , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate drug-resistant genes associated with beta-lactams and aminoglycosides in clinically isolated Pseudomonas aeruginosa.</p><p><b>METHODS</b>Twenty strains of Pseudomonas aeruginosa were isolated from wound excretion of hospitalized burn patients. The strains resistant to 14 antibiotics were selected for detection of 16 kind of drug-resistant genes (TEM, SHV, OXA-10 cluster, PER, VEB, GES, CARB, CTX-M- I, IMP, VIM, SPM, GIM, DHA, MOX, FOX, oprD2) and 6 kind of aminoglycoside modification genes (aac(3)- I, aac(3)-II, aac(6')-I, aac(6')-II, ant (3")- I , ant(2")- I) in them by PCR.</p><p><b>RESULTS</b>Among the 20 strains resistant to beta-lactam , all of them were TEM and GES positive (100%), oprD2 gene depletion in 5 strains (25%). All other genes were negative. Among aminoglycoside resistant genes, 20 strains were aac (6') - I positive (100%), 7 were ant (2") - I positive (35%), and negative for other stains.</p><p><b>CONCLUSION</b>There were very high existence rates of TEM, GES and aac (6')- I genes in Pseudomonas aeruginosa isolated from clinical burn patients. The fact that GES-5 gene has also been detected in Pseudomonas aeruginosa, suggesting this organism is highly drug resistant in our burn unit.</p>
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Humanos , Aminoglicosídeos , Farmacologia , Farmacorresistência Bacteriana Múltipla , Genética , Pseudomonas aeruginosa , Genética , Resistência beta-Lactâmica , Genética , beta-Lactamas , FarmacologiaRESUMO
Osteoporosis is a disease of bone metabolic disorder, the incidence of which increases sharply in old people. Calcitonin (CT) is a peptide hormone containing 32 amino acid that can inhibit osteoclasts activity. Osteogenic Growth Peptide (OGP) is a peptide hormone with 14 amino acid. It is an autocrine mitogen for osteoblastic and firbroblastic cells which has anabolic activiy. Six SCT and OGP DNA segments were chemically synthesized and ligated into a yeast expression vector pPIC9. The recombinant plasmid was transformed into pichia pastoris GS115. Finally, we got two stable SCT-OGP high expression clones after screening. Purifed protein can stimulate the proliferation of osteoblastic and fibroblastic cells, and also stimulate serum ALP activity and decrease serum calcium level using mice as animal models, demonstrating its potential role in oteoporosis therapy.
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Animais , Camundongos , Ratos , Calcitonina , Genética , Cálcio , Sangue , Histonas , Genética , Peptídeos e Proteínas de Sinalização Intercelular , Genética , Células NIH 3T3 , Osteoblastos , Osteoporose , Tratamento Farmacológico , Ratos Wistar , Proteínas Recombinantes de Fusão , Usos TerapêuticosRESUMO
<p><b>OBJECTIVE</b>To study on the dynamic changes of Codnopsis pilosula for the guidance of the field management.</p><p><b>METHOD</b>Using the random method the influences of the cultivating density and fertilizing weight at four different levels was observed.</p><p><b>RESULT AND CONCLUSION</b>The dynamic changes of the biomass, growth period andbiological changes at different growth stages were found out. The most predominant combination of the two factors, the 1.05 million roots/hm2 of the cultivating density and 240 kg x hm(-2) of the fertilizing weight reached the highest accumulation of stems & leaves; 652 mg x d(-1) the 0.6 million roots/hm2 and 240 kg x hm(-2) reached the heaviest single fresh root. The results could be used for the cultivating, fertilizing and practicing SOP of C. pilosula.</p>
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Biomassa , Codonopsis , Fertilizantes , Folhas de Planta , Raízes de Plantas , Caules de Planta , Plantas Medicinais , Estações do AnoRESUMO
<p><b>OBJECTIVE</b>To provide scientific basis for large scale production by studying the technique of tissue culture of Gentiana stramines.</p><p><b>METHOD</b>Callus was induced from germ-free stem segment of G. stramines on a MS medium supplemented with different hormones.</p><p><b>RESULT</b>The MS medium with 0.5-1.0 mg x L(-1) BA and 0.5 mg x L(-1) NAA was suitable for the induction and proliferation of cluster bads. MS medium with 1.0 mg x L(-1) IAA and 3 mg x L(-1) BA was suitable for the induction of calli. MS medium with 1.0 mg x L(-1) IAA and 2.0-3.0 mg x L(-1) BA was suitable for the subculture of calli. MS medium with 2.0 mg x L(-1) BA and 0.5 mg x L(-1) NAA was suitable for the differentiation of calli.</p><p><b>CONCLUSION</b>Aseptic seeding of G. stramines can be quickly propagated by shoot culture.</p>
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Meios de Cultura , Gentiana , Reguladores de Crescimento de Plantas , Farmacologia , Caules de Planta , Plantas Medicinais , Regeneração , Plântula , Técnicas de Cultura de Tecidos , MétodosRESUMO
<p><b>OBJECTIVE</b>To investigate the dynamic changes of the bacterial flora in our burn intensive care unit (ICU) in the past 6 years, and to analyze resistance of bacteria to various antibiotics.</p><p><b>METHODS</b>A retrospective analysis of bacterial culture and drug-sensitivity results from 209 patients in our burn intensive care unit during a period of 1998 to 2003 was carried out.</p><p><b>RESULTS</b>Eight hundred and forty-five strains of bacteria were isolated from 209 specimens, among which 486 strains were gram negative (G(-)) (57.51%), and 339 were gram positive (G(+)) (40.12%). Among all the G(+) bacteria, Enterococcus faecalis accounted for 34.51%, Staphylococcus aureus accounted for 31.27%, and 72.64% of Staphylococcus aureus strains were MRSA. Pseudomonas aeruginosa was predominant among all G(-) bacteria, and it accounted for 66.26% of the latter. All the bacteria isolated showed multiple resistance to antibiotics.</p><p><b>CONCLUSION</b>G(-) bacilli were still predominant in our burn intensive care unit. The isolated bacteria exhibited multiple resistance to antibiotics. The results imply that antibiotics should be administered rationally in the burn wards guided by the bacterial resistance test.</p>
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Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Antibacterianos , Farmacologia , Unidades de Queimados , Queimaduras , Tratamento Farmacológico , Microbiologia , Farmacorresistência Bacteriana Múltipla , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Testes de Sensibilidade Microbiana , Estudos RetrospectivosRESUMO
<p><b>OBJECTIVE</b>To analyze the isolation and the in vitro susceptibility of P. aeruginosa to antibiotics in our burn ward.</p><p><b>METHODS</b>Five hundred and thirty six burn patients admitted to our ward from 1997 to 2003 were enrolled in the study, and the wound excretion, the tips of the venous catheter, the subeschar tissue samples, and the blood samples were collected for bacterial identification and antibiotic susceptibility test with VITEK-AMS system.</p><p><b>RESULTS</b>The isolation rate of P. aeruginosa from 1997 to 2003 was 24.51%, 23.94%, 21.01%, 40.06%, 36.17%, 46.76% and 55.72%, respectively. The antibiotic effect of the third generation of Cephalosporins against the said bacteria showed a tendency to decline. The susceptibility rate to Cefoperazone, Ceftazidime and Cefotaxime were respectively 71%, 66% and 79% in 1997; 47%, 25%, 39% in 1998; 22%, 16%, 25% in 2002; The third generation cephalosporins had almost lost their antibiotic activity against P. aeruginosa in 2003, with the susceptibility rate to Cefotaxime lowered to 2%. The susceptibility rate to Imipenem from 1997 to 2003 was 76%, 33%, 45%, 11%, 41%, 31%and 4%, respectively.</p><p><b>CONCLUSION</b>The isolation rates of P. aeruginosa were steady during the period from 1997 to 1999, and they began to increase in 2000. The bacterial resistance to antibiotics increased gradually in recent years, and the strains of P. aeruginosa had become multi-drug resistant.</p>