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There is currently a huge worldwide demand for donor kidneys for organ transplantation. Consequently, numerous marginal donor kidneys, such as kidneys with microthrombi, are used to save patients' lives. While some studies have shown an association between the presence of microthrombi in donor kidneys and an increased risk for delayed graft function (DGF) (McCall et al., 2003; Gao et al., 2019), other studies have demonstrated that microthrombi negatively impact the rate of DGF (Batra et al., 2016; Hansen et al., 2018), but not graft survival rate (McCall et al., 2003; Batra et al., 2016; Gao et al., 2019). In contrast, Hansen et al. (2018) concluded that fibrin thrombi were not only associated with reduced graft function six months post-transplantation but also with increased graft loss within the first year of transplantation. On the other hand, Batra et al. (2016) found no significant differences in the DGF rate or one-year graft function between recipients in diffuse and focal microthrombi groups. To date, however, the overall influence of donor kidney microthrombi and the degree of influence on prognosis remain controversial, necessitating further research.
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Humans , Thrombotic Microangiopathies , Transplantation, Homologous , Tissue Donors , Kidney , AllograftsABSTRACT
Obesity is increasingly prevalent globally, searching for therapeutic agents acting on adipose tissue is of great importance. Equisetin (EQST), a meroterpenoid isolated from a marine sponge-derived fungus, has been reported to display antibacterial and antiviral activities. Here, we revealed that EQST displayed anti-obesity effects acting on adipose tissue through inhibiting adipogenesis in vitro and attenuating HFD-induced obesity in mice, doing so without affecting food intake, blood pressure or heart rate. We demonstrated that EQST inhibited the enzyme activity of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), a therapeutic target of obesity in adipose tissue. Anti-obesity properties of EQST were all offset by applying excessive 11β-HSD1's substrates and 11β-HSD1 inhibition through knockdown in vitro or 11β-HSD1 knockout in vivo. In the 11β-HSD1 bypass model constructed by adding excess 11β-HSD1 products, EQST's anti-obesity effects disappeared. Furthermore, EQST directly bond to 11β-HSD1 protein and presented remarkable better intensity on 11β-HSD1 inhibition and better efficacy on anti-obesity than known 11β-HSD1 inhibitor. Therefore, EQST can be developed into anti-obesity candidate compound, and this study may provide more clues for developing higher effective 11β-HSD1 inhibitors.
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Equisetin (EQST) belongs to polyketide (PKS)-nonribosomal peptide synthetase (NRPS) type compound with an inhibitory effect of 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) enzyme activity. This study investigated anti-obesity effect and insulin resistance improvement effect of EQST on high-fat diet (HFD)-induced ob/ob mice model. EQST treatment effectively reduced the body weight gain, fat weight gain and blood lipid content of model mice. All animal experiments were approved by the Medical Ethics Committee of Capital Institute of Pediatrics. EQST alleviated adipose tissue expansion and hepatic ballooning degeneration of model mice, and also effectively controlled the blood glucose content after glucose load and insulin load, showed a significant improvement in obesity and insulin resistance. EQST inhibited adipogenic proteins fatty acid-binding protein 4 (FABP4) and peroxisome proliferators-activated receptor γ (PPARγ), and upregulated thermogenic protein (uncoupling protein 1, UCP1) through suppressing 11β-HSD1 protein expression. In addition, EQST widely upregulates mitochondrial respiratory metabolism related proteins in adipose tissue and may improve insulin resistance through phosphatidylinositol-3-kinase (PI3K) pathway. Therefore, EQST plays an anti-obesity role by promoting adipose tissue thermogenesis and improving insulin resistance, which may provide reliable clues for improving obesity and diabetes.
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Objective:To rapidly identify the chemical constituents in Xiao Chengqitang by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS）. Method:The method was established by the Waters CORTECS T3 column （2.1 mm×150 mm， 1.6 μm）， mobile phase was methanol （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-5 min， 3%-21%A； 5-20 min， 21%-36%A； 20-32 min， 36%-50%A； 32-42 min， 50%-62%A； 42-50 min， 62%-85%A； 50-60 min， 85%-95%A）， the flow rate was 0.2 mL·min<sup>-1</sup>， and the column temperature was 30 ℃. UPLC-Q-Orbitrap-MS was operated in positive and negative ion modes， the scanning range was 100-1 200 with mode of Full MS/dd-MS<sup>2</sup>， and the collision energies were 20， 40 eV. The compounds were identified by comparing with reference substances and combining with literature reports and MS database information. Result:A total of 123 components were identified in Xiao Chengqitang， including 33 flavonoids， 25 anthraquinones and anthrones， 23 phenylpropanoids， 15 tannins， 10 nitrogen-containing components and 17 other components. Among them， 32 components were determined by reference substances. Conclusion:The material basis of Xiao Chengqitang is flavonoids， anthraquinones and anthrones， phenylpropanoids， which is derived from Aurantii Fructus Immaturus，<italic> </italic>Rhei Radix et Rhizoma and Magnoliae Officinalis Cortex， respectively.
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Respiratory training can prevent and treat nonspecific low back pain, mainly by activating the deep stabilizing muscles of trunk, establishing appropriate intra-abdominal pressure and optimizing the proprioceptive input of lumbar back, so as to maintain the stability and control of the spine. At present, the commonly used breathing training methods in clinic mainly include diaphragmatic breathing, inspiratory muscle training and abdominal breathing. However, the formulation of respiratory training prescriptions and the comparative study of therapeutic effects among different respiratory training methods still need to be further explored.
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Objective:To analyze the long-term efficacy and safety of re-irradiation for recurrent glioma.Methods:The data of 52 patients with recurrent gliomas were collected from 2009 to 2019. The median planned targetvolume (PTV) was 73.5 cm 3(49.9-102.7 cm 3) and the median dose was 45.0 Gy (43.0-48.8 Gy). Kaplan-Meier method was used for survival assessment, log-rank test for difference assessment, and Cox’s regression model for multivariate prognostic analysis. Results:The median follow-up time was 32.6 months. The median overall survival (OS) and progression-free survival (PFS) time were 16.1 months (95% CI, 4.1-28.1) and 8.0 months (95% CI, 4.0-12.0). The 1-, 2-and 3-year survival rates were 67%, 43% and 29%, respectively. The 6-month, 1-year and 2-year PFS rates were 67%, 40%, 26%, respectively. Multivariate analysis showed that KPS score and recurrence time significantly affected the OS ( P=0.012, P=0.001). KPS score and time interval between two radiotherapies significantly impacted the PFS ( P=0.003, P=0.018). Stratified analysis showed that KPS score was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅱ initial pathology and reoperation after recurrence ( P<0.001, P=0.012); clinical manifestation was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅲ and Ⅳ initial pathology ( P=0.006, P=0.044). The overall incidence of adverse reactions was 30.8%. Grade 1 adverse reactions accounted for 25.0%, and 5.8% for grade 2. Conclusions:Re-irradiation for recurrent glioma yields good long-term clinical efficacy and tolerable adverse reactions.
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As an interdisciplinary subject of medicine and artificial intelligence, intelligent diagnosis and treatment has received extensive attention. The standardization of Chinese medicine (CM) diagnosis has been always a bottleneck in the modernization and globalization of CM. Studying the application technology of artificial intelligence in CM and solving the problems is an urgent need for the development of modern CM in the era of artificial intelligence. Taking the pneumonia with dyspnea and cough in CM as an example, this article gives an overview of intelligent medical technology and application development, brings forward the present technical problems faced and the new advances in intelligent technology on CM diagnosis and treatment.
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Gentianae Radix et Rhizoma has a long history of medication,and was recorded in Shennong Bencaojing. Through the study of herbal medicine of Gentianae Radix et Rhizoma,we reviewed and analyzed previous herbal records,studied its basic,origin and processing methods, and provided the basis for further development and utilization of Gentianae Radix et Rhizoma. Through the analysis of ancient herbal researches and drawings,we knew that the ancient resources of this medicine were basically consistent with Gentiana scabra and G. manshurica. The Latin name of the original plant of gentian medicinal herbs set forth in the modern literature was analyzed for the first time. The commodities of this medicine were divided into Guanlongdan and Dianlongdan. Guanlongdan was produced in Northeast China, and consisted of G. scabra,G. manshurica,and G. triflora. Dianlongdan was G. rigescens and grew in Yunnan province. Due to various reasons, such as historical changes, genuine producing areas of medicinal materials gradually changed from Jiangsu and Zhejiang to Northeast China. The processing methods of Gentianae Radix et Rhizoma included preparing with wine, preparing with licorice juice and preparing with ginger juice. The purpose of these processing methods is to suppress its nature of bitter and cold. Among those processing methods,preparing with wine has the longest history, and can promote the dissolution of the active ingredients of gentian,thus is a better method of processing. This study provides the basis for radical reform, resource development, correct use, and determination of genuine producing area and processing methods for Gentianae Radix et Rhizoma.
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<p><b>Background</b>Acute coronary syndrome (ACS) is closely related to unstable plaques and secondary thrombosis. The inflammatory cells in plaques and their inflammatory products may be the cause for plaque instability and ruptures. The study aimed to disclose the changes of inflammatory factors including serum intracellular adhesion molecule-1 (ICAM-1), chitinase-3-like protein 1 (YKL-40), and lipoprotein-associated phospholipase A2 (Lp-PLA2) in patients with ACS and its clinical significance.</p><p><b>Methods</b>A total of 120 patients with coronary heart disease (CHD) were categorized into 2 groups: 69 with ACS and 51 with stable angina pectoris (SAP); 20 patients with chest pain and normal angiography served as a control group. The 120 patients with CHD were categorized into single-vessel disease group, double-vessel disease group, and three-vessel disease group based on the number of coronary artery stenosis. The severity of coronary artery stenosis was quantified based on coronary angiography using Gensini score. They were further divided into mild CHD group with its Gensini score <26 (n = 36), moderate CHD group with its Gensini score being 26-54 (n = 48) and severe CHD group with its Gensini score >54 (n = 36). Serum levels of ICAM-1, YKL-40, and Lp-PLA2 of different groups were determined by enzyme-linked immunosorbent assay. Correlation between ICAM-1, YKL-40, Lp-PLA2, and Gensini score was analyzed.</p><p><b>Results</b>The levels of serum inflammatory factors ICAM-1, YKL-40, and Lp-PLA2 were significantly higher in the ACS group than those in control group and SAP group (all P < 0.05); and compared with control group, no significant difference was observed in terms of the serum ICAM-1, YKL-40, and Lp-PLA2 levels in the SAP group (P > 0.05).The levels of serum ICAM-1, YKL-40, and Lp-PLA2 were not significantly different among control group, single-vessel disease group, double-vessel disease group, and three-vessel disease group (all P > 0.05). The levels of serum ICAM-1, YKL-40, and Lp-PLA2 were not significantly different among control group, mild CHD group (Gensini score <26), moderate CHD group (Gensini score 26-54), and severe CHD group (Gensini score >54) (all P > 0.05). Nonparametric Spearman correlation analysis showed that the levels of serum ICAM-1, YKL-40, and Lp-PLA2 were not correlated with the Gensini score in CHD patients (r = 0.093, r = -0.149, and r = -0.085, all P > 0.05; respectively).</p><p><b>Conclusions</b>The serum levels of ICAM-1, YKL-40, and Lp-PLA2 were correlated with different clinical types of CHD, but not well correlated the severity and extent of artery stenosis, suggesting that ICAM-1, YKL-40, and Lp-PLA2 might be involved in occurrence of instability of atherosclerotic plaque, and might reflect the severity of CHD mostly through reflecting the plaque stability.</p>
Subject(s)
Adult , Aged , Humans , Middle Aged , 1-Alkyl-2-acetylglycerophosphocholine Esterase , Metabolism , Acute Coronary Syndrome , Blood , Allergy and Immunology , Metabolism , Chitinase-3-Like Protein 1 , Metabolism , Coronary Angiography , Coronary Disease , Blood , Allergy and Immunology , Metabolism , Intercellular Adhesion Molecule-1 , MetabolismABSTRACT
Objective To compare the clinical effect between percutaneous nephrolithotomy (PCNL) and retroperitoneal laparoscopic pyelolithotomy (RLP) in the treatment of renal pelvis calculus,so as to provide evidence for the treatment of renal pelvis calculus.Methods A total of 108 patients with renal pelvis calculus were selected from January 2012 to December 2016 in Zhumadian Central Hospital.The patients were divided into PCNL group (n =59) and RLP group (n =49) according to the therapeutic method.The operation time,intraoperative blood loss,intraoperative blood transfusion rate,operative success rate,stone clearance rate,postoperative hemoglobin reduction,postoperative analgesia time,postoperative hospitalization time and the incidence of complications were compared between the two groups.Results The success rate of operation in PC-NL group and RLP group was 94.9％ (56/59) and 100.0％ (49/49) respectively,there was no significant difference in the success rate of operation between the two groups (x2 =1.026,P ＞ 0.05).The stone clearance rate in PCNL group and RLP group was 94.9％ (56/59) and 98.0％ (48/49) respectively,there was no significant difference in the stone clearance rate between the two groups (x2 =0.140,P ＞ 0.05).The intraoperative blood transfusion rate in PCNL group and RLP group was 5.1％ (3/59) and 2.0％ (1/49) respectively,there was no significant difference in the intraoperative blood transfusion rate between the two groups (x2 =0.105,P ＞ 0.05).There was no significant difference in operation time and postoperative analgesia time between the two groups (P ＞ 0.05).Compared with the PCNL group,the blood loss and postoperative hemoglobin reduction were less,and the postoperative hospitalization time was shorter in the RLP group (P ＜ 0.05).The incidence of postoperative urinary leakage,urinary tract infection and secondary hemorrhage in PCNL group was 3.4％ (2/59),8.5％ (5/59)and 6.8％ (4/59) respectively;the incidence of postoperative urinary leakage,urinary tract infection and secondary hemorrhage in RLP group was 8.2％ (4/49),4.1％ (2/49) and 4.1％ (2/49) respectively;there was no significant difference in the incidence of postoperative urinary leakage,urinary tract infection and secondary hemorrhage between the two groups (x2 =2.975,1.064,1.811;P ＞ 0.05).Conclusion The clinical effect of PCNL and RLP in the treatment of renal pelvis calculus is fairly,and their safety is high.However,RLP has the advantages of less intraoperative bleeding,quick postoperative recovery and short hospitalization time.
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This work aims to analyZe and compare the potential biomarkers between cervical cancer ( CC ) patients and healthy controls (HC).The urine samples of 11 CC patients (age (45.65 ± 5.6) years) and 11 HC patients (age (45.9 ± 3.2) years) were collected.The metabolites of urine samples from CC and HC were analyZed by using ultra performance liquid chromatography coupled with quadrupole orbitrap tandem mass spectrometry ( UHPLC-Q-Orbitrap MS/MS ) , which could provide evidence for early diagnose and disease pathway.The LC-MS data of urines were analyZed by principal components analysis ( PCA) and orthogonal partial least squares discriminant analysis ( OPLS-DA) to identify the potential biomarkers.Urine samples of CC patients were successfully discirminated from those of healthy controls.A total of twelve significant metabolites were found and identified as potential biomarkers according to the established UHPLC-Q-Orbitrap MS and MS/MS method.Identification of biomarkers between CC and HC patients may play an important role in the study of mechanism of UC and its pathway.
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Objective:To compare the long-term prognosis of percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG) in patients with ostial/shaft lesions of unprotected left main coronary artery (ULMCA).Methods:A total of 259 patients with isolated ostial/midshaft lesions of ULMCA who received PCI or CABG in our hospital from 2003-01 to 2009-07 were enrolled.The patients were divided into 2 groups:PCI group,n=149 patients who received drug-eluting stents (DES) implantation and CABG group,n=110.The end points were all cause death,myocardial infarction (MI) and major adverse cardiac and cerebrovascular events (MACCE) which included cardiac death,non-fetal MI,stroke,repeated revascularization and the composite events death.Results:The median follow-up period was 7.1 years (inter quartile range 5.3-8.2 years) in all patients.Before multivariate adjusting,the following parameters were similar between PCI group and CABG group:all cause death (12.7％ vs 29.7％),P=-0.096;non-fatal MI (14.8％ vs 8.5％),P=0.844;stroke (9.3％ vs 6.3％),P=0.904;repeated revascularization (26.8％ vs 19.0％),P=0.234;composite events of cardiac death/stroke/MI (18.9％ vs 20.3％),P=0.224 and MACCE occurrence (37.5％ vs 34.2％),P=0.946.With adjusted variations,the trend was similar to pre-adjustment.Conclusion:During the maximum 8.2 years follow-up period,PCI and CABG had the similar efficacy and safety in patients with ostial/shaft lesions of ULMCA.
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Objective:To compare the long-term prognosis of percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG) in patients with ostial/shaft lesions of unprotected left main coronary artery (ULMCA).Methods:A total of 259 patients with isolated ostial/midshaft lesions of ULMCA who received PCI or CABG in our hospital from 2003-01 to 2009-07 were enrolled.The patients were divided into 2 groups:PCI group,n=149 patients who received drug-eluting stents (DES) implantation and CABG group,n=110.The end points were all cause death,myocardial infarction (MI) and major adverse cardiac and cerebrovascular events (MACCE) which included cardiac death,non-fetal MI,stroke,repeated revascularization and the composite events death.Results:The median follow-up period was 7.1 years (inter quartile range 5.3-8.2 years) in all patients.Before multivariate adjusting,the following parameters were similar between PCI group and CABG group:all cause death (12.7％ vs 29.7％),P=-0.096;non-fatal MI (14.8％ vs 8.5％),P=0.844;stroke (9.3％ vs 6.3％),P=0.904;repeated revascularization (26.8％ vs 19.0％),P=0.234;composite events of cardiac death/stroke/MI (18.9％ vs 20.3％),P=0.224 and MACCE occurrence (37.5％ vs 34.2％),P=0.946.With adjusted variations,the trend was similar to pre-adjustment.Conclusion:During the maximum 8.2 years follow-up period,PCI and CABG had the similar efficacy and safety in patients with ostial/shaft lesions of ULMCA.
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<p><b>OBJECTIVE</b>To investigate the relationship between inflammatory factors and two Chinese medicine (CM) syndrome types of qi stagnation and blood stasis (QSBS) and qi deficiency and blood stasis (QDBS) in patients with acute coronary syndrome (ACS).</p><p><b>METHODS</b>Sixty subjects with ACS, whose pathogenesis changes belongs to qi disturbance blood stasis syndrome, were divided into 2 groups: 30 in the QSBS group and 30 in the QDBS group. The comparative analysis on them was carried out through comparing general information, coronary angiography and inflammatory factors including intracellular adhesion molecule-1 (ICAM-1), chitinase-3-like protein 1 (YKL-40) and lipoprotein-associated phospholipase A2 (Lp-PLA2).</p><p><b>RESULTS</b>Compared with the QSBS group, Lp-PLA2 and YKL-40 levels in the QDBS group showed no-significant difference (P>0.05); ICAM-1 was significantly higher in the QDBS group than in the QSBS group in the pathological processes of qi disturbance and blood stasis syndrome of ACS (P<0.05).</p><p><b>CONCLUSIONS</b>Inflammatory factor ICAM-1 may be an objective basis for syndrome typing of QSBS and QDBS, which provides a research direction for standardization research of CM syndrome types.</p>
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Acute kidney injury (AKI) is a common complication following orthotopic liver transplantation (OLT) and is associated with increased morbidity and mortality.The aim of the current study was to determine the risk factors for AKI in patients undergoing OLT.A total of 103 patients who received OLT between January 2015 and May 2016 in Tongji Hospital,China,were retrospectively analyzed.Their demographic characteristics and perioperative parameters were collected,and AKI was diagnosed using 2012 Kidney Disease:Improving Global Outcomes (KDIGO) staging criteria.It was found that the incidence of AKI was 40.8％ in this cohort and AKI was significantly associated with body mass index,urine volume,operation duration (especially ＞ 480 min),and the postoperative use of vasopressors.It was concluded that relative low urine output,long operation duration,and the postoperative use of vasopressors are risk factors for AKI following OLT.
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Adenoid hypertrophy is a disease that mostly occurs among children of 3-5 years old. It is caused by repeated inflammation and infection of nasopharynx and its adjoin parts, or the adenoid itself, which will finally leads to pathological hyperplasia of adenoid. With so much information we have acquired about this disease, its specific mechanism remains unknown. In recent years, some researches have indicated that adenoid hypertrophy may have something to do with extra-gastroesophageal reflux, in which pepsin plays a very important role, and pepsin will do a series of pathological damages to the upper airway as it reaches the upper respiratory tract. Based on relative domestic and foreign literature, this paper attempts to make a review about the relationship between gastroesophageal reflux and adenoid hypertrophy.
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Child , Humans , Adenoids , Pathology , Gastroesophageal Reflux , Hypertrophy , Nasopharynx , Pathology , Pepsin A , MetabolismABSTRACT
Objective To investigate the influencing factors and improve potency methods of chymosin, to verify the stability and applicability of the national standard of chymosin.Methods The effects of different formula milk powder substrate and enzyme concentration on the determination of the activity of chymosin were studied.3 ×3 dose-response parallel line method was established.The results were compared with the different methods of absolute and relative methods.Results The different formula milk powder had a significant effect on the determination of the absolute potency of the activity of chymosin.The concentration of the enzyme was a power function relationship with the milk clotting time.Compared with the absolute potency, reproducibility of the relative potency of the results was better in different laboratories.The suitable doses in 3 ×3 dose-response parallel line method were 0.35,0.44,0.55U/mL.The confidence limit rate was less than 5%.The potency of the national standard of chymosin (140712-201302) was not significantly different between 2013 and 2015.In a certain dose range, the dose-response of the national standard of chimosin and gastropylor complex or lamb'tripe extract was linear, and the two lines were parallel.Conclusion A lot of factors can affect on the potency of chymosin.Relative potency is determinate by reference standard which can eliminate the influence of different substrates, different operators and endpoint judgment on the determination in order to make results have comparability between laboratories.The test design of 3 ×3 dose-response parallel line can control the test deviation better than the single point determination.The stability of the national standard of chymosin(140712-201302) is good, and is suitable for the potency of chymosin of the products of gastropylor complex and the extract of the lamb.
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Objective To improve the related substances analysis method of cobamamide.And to further research light degradation characteristics of cobamamide.Methods Determined the related substances of solid and aqueous solution of cobamamide after light degradation by high performance liquid chromatography(HPLC) conditions,which is Merck Hibar C18 column (4.6 mm ×250 mm, 5μm), 0.05 mol/L KH2PO4 solution:acetonitrile (90:10) -80% acetonitrile as mobile by gradient elution and detection wavelength 260 nm.And compared their light stability.The main three kinds of light degradation impurities were determinate from LC-MS.Results Gradient elution made the light degradation impurities separate better.The results of precision and reproducibility tests increased to RSD =0.2% (n=6) and RSD =8% (n=5) from RSD =3.8% (n =6) and RSD=38%(n=5). Cobamamide solution was very sensitive to light, the preparation should be strict dark operation.Two of the light degradation impurities were adenosine and hydroxycobalamin, with the relative response factor 2.5 and 0.7.Conclusion New method is specific, durable and reproducible, which can be used for quality control of cobamamide.
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OBJECTIVE: To establish an LC-MS method for identification of the primary degradation impurities in domestic calcitonin salmon injections. METHODS: For the RPLC analysis., the separation was performed on Waters XTerra RP C18 column (3.0 mm × 50 mm, 3.5 μm) with linear gradient elution using 0.05% aqueous trifluoroacetic acid (A) and acetonitrile (B); the flow rate was 0.5 mL · min-1; the column temperature was 40℃; the detection wavelength was set at 220 nm. For the SEC analysis, the separation was performed on TSK GEL 2000SWxl column (7.8 mm × 300 mm, 5 μm) with mobile phase consisting of TFA-water-acetonitrile(0.05:80:20); the flow rate was 0.7 mL · min-1; the column temperature was room temperature; the detection wavelength was set at 220 nm; the main impurities were collected and identified by positive mode. RESULTS: The structures of three impurities were elucidated by RPLC-MS and SEC-MS. CONCLUSION: The results of this study can be applied in impurity profile analysis, specification research, and quality control of calcitonin salmon injections.
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Objective To validate the determination method of microbacteria limit of Shuitiaosan powder .Methods Plate counting method was used .The method of counting bacteria and mould was validated by the recovery rates with 5 control strains .The method of checking control bacteria was validated by observing cultivation of Staphylococcus aureus and Pseudomonas aeruginosa in the test group, positive control group and negative control group in the same environment .Results The recovery rate of every trail strains was higher than 70%when centrifugal sedimentation methods were used in the counting bacteria and mould .To the examination of con-trol bacteria , Staphylococcus aureus and Pseudomonas aeruginosa were detected by the centrifugal sedimentation methods .The tested strains were observed in the test group and in the positive control group .No strains were observed in the negative control group .Con-clusion The methods are simple , feasible, reliable and can be used for the examination of microbacteria limit .