Exposure to second-hand smoke is an independent risk factor of small airway dysfunction in non-smokers with chronic cough: A retrospective case-control study.

Objectives: This study aimed to identify the potential risk factors for small airway dysfunction (SAD) in non-smokers with chronic cough. Methods: Non-smokers with chronic cough who underwent lung function tests at Xiangya Hospital from May 2019 to May 2020 were enrolled, and divided into the derivation and validation cohorts based on their hospital admission time. SAD was determined based on the presence of at least two of the following three indicators of lung function being less than 65% of predicted: maximal mid-expiratory flow, forced expiratory flow at 50% of forced vital capacity (FVC), and forced expiratory flow at 75% of FVC. Clinical data of these patients were collected. Risk factors for SAD were identified by logistic regression analysis in the derivation cohort and further confirmed in the validation cohort. Results: In total, 316 patients (152 in the non-SAD group and 164 in the SAD group) were included in the derivation cohort. Compared with the non-SAD group, the SAD group had a higher proportion of female patients (82.3 vs. 59.2%, P < 0.001), was more commonly exposed to second-hand smoke (SHS) (61.6 vs. 27.6%, P < 0.001), and tended to be older (median age, 45.5 vs. 40.0 years old, P = 0.004). The median FVC, forced expiratory volume in one second (FEV1) % pred, FEV1/FVC ratio, and peak expiratory flow (PEF) % pred were slightly lower in the SAD group. Multivariable logistic analysis showed that exposure to SHS was an independent risk factor (OR 4.166 [95% CI 2.090-8.302], P < 0.001) for SAD in non-smokers with chronic cough after adjusting for related variables. In the validation cohort (n = 146), patients with SHS exposure had a relative risk of 1.976 (95% CI 1.246-3.135, P = 0.004) for SAD compared to those without SHS exposure. Multivariable logistic analysis consistently confirmed that exposure to SHS was an independent risk factor (OR 3.041 [95% CI 1.458-6.344], P = 0.003) for SAD in non-smokers. Conclusions: Exposure to SHS is independently associated with a higher risk of SAD in non-smokers with chronic cough. Reduction in SHS exposure may ameliorate lung function, thus lowering the risk of irreversible airway obstruction.

A network pharmacology-based study on the mechanism of astragaloside IV alleviating renal fibrosis through the AKT1/GSK-3ß pathway.

ETHNOPHARMACOLOGICAL REVELVANCE: Astragaloside IV, a glycoside derived from Astragalus membranaceus, has anti-renal fibrosis effects. However, its mechanism of action has not yet been fully elucidated. AIM OF THE STUDY: The purpose of this study was to investigate the anti-fibrotic effect of AS-IV and to clarify its underlying mechanism. MATERIALS AND METHODS: The network pharmacology method, molecular docking and surface plasmon resonance (SPR) was used to identify potential targets and pathways of AS-IV. A unilateral ischemia-reperfusion injury (UIRI) animal model, as well as TGF-ß1-induced rat renal tubular epithelial cells (NRK-52E) and renal fibroblasts (NRK-49F) were used to investigate and validate the anti-fibrotic activity and pharmacological mechanism of AS-IV. Network pharmacology was performed to construct a drug-target-pathway network. The anti-fibrosis effect of AS-IV was determined using hematoxylin-eosin (H&E) and MASSON staining, as well as immunostaining methods. qRT-PCR and western blotting were used to elucidate and validate the mechanism of AS-IV. RESULTS: Network pharmacology revealed that the PI3K/AKT pathway is an important pathway in AS-IV. AS-IV inhibited the expression of α-SMA, collagen I, and fibronectin in NRK-52E, NRK-49F, and UIRI rats, and reduced serum creatinine and blood urea nitrogen levels in UIRI rats. AS-IV inhibited AKT phosphorylation, blocked GSK-3ß phosphorylation, and restored GSK-3ß activity, which contributed to the degradation of ß-catenin, thereby preventing epithelial-mesenchymal transition (EMT). CONCLUSION: Astragaloside IV alleviated renal fibrosis through the AKT1/GSK-3ß pathway. In addition, our findings indicate that the network pharmacology method is a powerful tool for exploring the pharmacological mechanisms of drugs.

An integrated network pharmacology and cell metabolomics approach to reveal the role of rhein, a novel PPARα agonist, against renal fibrosis by activating the PPARα-CPT1A axis.

BACKGROUND: Rhein, an anthraquinone compound, displays extensive antifibrotic effects; however, its potential mechanisms are not fully understood. In this study, we explored the underlying molecular mechanism of action of rhein. METHOD: An integrated network pharmacology and cell metabolomics approach was developed based on network pharmacology and bioinformatics method, and then successfully applied to speculate the potential targets of rhein and construct a rhein-target-metabolic enzyme-metabolite network. Thereafter, the antifibrotic mechanism of rhein was validated in TGF-ß- and oleic acid- induced HK-2 and NRK-52E cells in vitro as well as a unilateral ischemia-reperfusion injury Sprague-Dawley rat model. RESULTS: Based on the construction of the rhein-target-metabolic enzyme-metabolite network, we found that rhein played an antifibrotic role through the PPAR-α-CPT1A-l-palmitoyl-carnitine axis. In vitro experiments demonstrated that rhein effectively activated the expression of PPARα and its downstream proteins (CPT1A and ACOX1) to alleviate lipid accumulation and fibrosis development. In vivo experiments indicated that rhein attenuated renal fibrosis mainly by activating the fatty acid oxidation pathway and improving lipid metabolism. CONCLUSION: Taken together, our findings reveal that rhein is a novel agonist of PPARα, which contributes to its renoprotection through the regulation of the PPARα-CPT1A axis. Moreover, our study provides a novel insight into an integrated network pharmacology-metabolomics strategy for uncovering the pharmacological mechanisms of drugs from the system perspective.

GC-MS/MS method for determination and pharmacokinetics of sclareol in rat plasma after intravenous administration.

Sclareol, mainly isolated from Salvia officinalis, has a variety of pharmacological effects. In this work, a sensitive and rapid gas chromatography-tandem mass spectrometry (GC-MS/MS) method was first developed and validated for the determination and pharmacokinetics of sclareol in rat plasma. The chromatographic seperation of biosamples was performed with an HP-5MS column. Ethyl acetate was selected as the solvent to extract analytes from rat plasma. The multiple reaction monitoring transitions for sclareol and dehydrocostuslactone (Internal Standard, IS) were m/z 177 â†’ 121 and m/z 230 â†’ 173, respectively. The intra- and inter- precision, accuracy, matrix effect, recovery and stability meet the method requirements for biological sample analysis. The lowest limit of quantification (LLOQ) of the developed method for sclareol determination was 20 ng/mL. After intravenous administration (5.0 mg/kg) of sclareol to the rats, its drug clearance (CLz) and elimination half-life (t1/2z) was 2.7 ± 1.3 L/h/kg and 6.0 ± 4.6 h, respectively. The apparent volume of distribution (Vz) was 21.4 ± 12.9 L/kg, which indicated that sclareol was mainly distributed in extracellular fluid. Our results provided useful information for the further pharmacological investigation and preclinical studies of sclareol.

Chemical constituents of radix Actinidia chinensis planch by UPLC-QTOF-MS.

Radix Actinidia decoction and its prescriptions are used to treat tumors and other diseases. Although some chemical components have been isolated from Radix Actinidia, systematic analysis of its chemical components has not been reported, which hinders the basic research on its effective substances and its quality control. In this work, a UPLC-QTOF-MS method was employed to profile and characterize the chemical constituents of water extracts from Radix Actinidia Chinensis Planch (RACP). We unambiguously or tentatively identified 295 chemical components from RACP, including 46 pentacyclic triterpenes, 72 flavonoids, 53 phenolic acids, 24 coumarins, three anthraquinones and other compounds. Most of the chemical components have not been described so far in Actinidia. More than 180 phytochemicals are reported in Actinidia for the first time. 2α,3α,24-trihydroxyurs-12-en-28-oic acid, asiatic acid, syringic acid, fraxin, esculetin, 5,7-dihydroxychromone, esculin, (+)-catechin, (-)-epi-catechin, vanillic acid, ferulic acid, protocatechuic acid and rutin were unambiguously identified by comparison with the reference standards. Catechin derivatives, coumarin derivatives and phenolic acid derivatives were the main water-soluble components in RACP. This study broadened the chemical profiles of RACP, and laid the foundation for subsequent research on the effective components and their mechanism of action. This work also provides an important reference for the quality control and evaluation of RACP.

Pharmacokinetics and Pharmacodynamics of the Combination of Rhein and Curcumin in the Treatment of Chronic Kidney Disease in Rats.

Objectives: The interaction between the components of traditional Chinese medicine (TCM) is an important basis for their synergy. Rhein and curcumin exert various pharmacological activities, including anti-tumour, anti-inflammatory, antioxidant, anti-fibrosis and renoprotective effects. However, no investigation has reported the synergistic anti-fibrosis effect yet. This study aims at determine the pharmacokinetics and pharmacodynamics of the combination of rhein and curcumin in the treatment for chronic kidney disease in rats. Design: Fifty two male Sprague-Dawley (SD) rats were randomly divided into rhein group, curcumin group and their combination group for pharmacodynamics studies. HE and Masson staining was conducted to observe the changes of renal morphology. Kits were used to detect the level of urea nitrogen (BUN) and creatinine (Scr). For pharmacokinetic study, 36 SD rats were randomly divided into rhein group, curcumin group and a combination group, the content of rhein and curcumin in plasma and renal tissue was determined by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). In additon, molecular docking method and cell experiments was used to disclose the interaction mechanism between curcumin and rhein. Results: The pharmacodynamic results showed that the degree of renal fibrosis was improved obviously by co-administration rhein and curcumin. Meanwhile, compared to single administration, the Cmax and AUC of rhein and curcumin in plasma and renal tissue were enhanced significantly after co-administration. Moreover, the result of molecular docking and cell experiments showed that both two compounds could interact with P-gp, CYP2C9 and CYP2C19. Conclusion: Together, these findings demonstrated that rhein and curcumin had a synergistic effect in ameliorateing chonic kidney disease, providing an important explanation on the synergistic mechanism of curcumin and rhein from a pharmacokinetic viewpoint.

Long non-coding RNA TCF7 contributes to the growth and migration of airway smooth muscle cells in asthma through targeting TIMMDC1/Akt axis.

BACKGROUND: Long noncoding RNAs (lncRNAs) have been revealed to participate in cellular biological processes in multiple diseases, including asthma. Nevertheless, the role of lncRNA TCF7 (lncTCF7) in airway smooth muscle cells (ASMCs) is still covered. METHODS: The expression of lncTCF7 and TIMMDC1 in ASMCs from 12 asthma patients and 12 healthy controls were detected using qRT-PCR. Then MTT assay, EdU assay and transwell assay were conducted respectively to assess the impact of lncTCF7 on ASMCs viability, proliferation and migration. Besides, western blotting was performed to determine the protein levels of TIMMDC1 and AKT/p-AKT. RESULTS: We discovered that lncTCF7 and TIMMDC1 were upregulated in asthma groups and lncTCF7 improved ASMCs viability/proliferation and migration. In addition, lncTCF7 regulated TIMMDC1 expression indeed and PDGF-BB treated ASMCs exhibited elevated levels of lncTCF7 and TIMMDC1. Moreover, lncTCF7 suppression diminished both the mRNA and protein levels of TIMMDC1 and markedly reduced p-AKT level which could be enhanced under TIMMDC1 overexpression. Finally, both TIMMDC1 overexpression and AKT activator could restored the inhibitory impacts of lncTCF7 silence on PDGF-BB treated ASMCs. CONCLUSION: Our study uncovered that lncTCF7 facilitated human ASMCs growth and migration via targeting TIMMDC1 thus activating AKT signaling, providing a novel possible target for asthma therapy.

Multinodule abnormalities of the tracheobronchus: bronchoscopy findings and clinical diagnosis.

BACKGROUND AND AIMS: Bronchoscopy is an important method for diagnosing respiratory disease. Multiple tracheobronchial nodules are rarely reported and their causes remain unclear. OBJECTIVES: The aim of this study was to describe the clinical characteristics of multiple nodule tracheobronchial abnormalities found under bronchoscopy caused by different diseases. METHODS: Eighty-seven patients with multiple tracheobronchial nodules were enrolled in this study. The characteristics of the multinodule lesions and the patient were diagnosed based on the pathology findings in our hospital. Chest computed tomography images were retrospectively reviewed by pulmonologists and radiologist. RESULTS: In 55 patients with definite pathological diagnosis, 16 (29%) patients were diagnosed as tuberculosis (TB) granuloma; 23 (41.8%) cases were diagnosed as malignant disease; 12 (21.8%) cases were diagnosed as tracheobronchopathia osteochondroplastica; 2 (3.6%) cases were diagnosed as sarcoidosis; and one case (1.8%) was diagnosed as lymphoma and one case (1.8%) as fungal infection. There were 32 cases of chronic inflammation. There was no relationship between nodule distribution and the pathological diagnosis. Malignant nodules usually smaller with a pale outlook, while nodules with larger size and smooth and intact mucosa usually turn out to be granuloma of unknown reason. CONCLUSION: The major causes of mutinodule lesions observed using bronchoscopy are tumor and TB. The presence of multiple endotracheobronchial nodules suggest that pulmonary lesion is present, and biopsy should be performed. Malignant nodules can be diagnosed by appearance and biopsy. Pathology results of TB, sarcoidosis and fungal infection can turn out to be granuloma of unknown reason. Further diagnosis needs other clinical materials.

Fluorofenidone attenuates TGF-ß1-induced lung fibroblast activation via restoring the expression of caveolin-1.

Caveolin-1 plays an important role in the pathogenesis of idiopathic pulmonary fibrosis. We previously showed that fluorofenidone (FD), a novel pyridine agent, can attenuate bleomycin-induced experimental pulmonary fibrosis and restore the production of caveolin-1. In this study, we explore mainly whether caveolin-1 plays a critical role in the anti-pulmonary fibrosis effects of FD in vitro. The normal human lung fibroblasts (NHLFs) were cultured with transforming growth factor-ß1 (TGF-ß1) and then were treated with FD. Subsequently, NHLFs transfected with cav-1-siRNA were treated with TGF-ß1 and/or FD. The expressions of α-smooth muscle actin (α-SMA), fibronectin, collagen I, caveolin-1, phosphorylated extracellular signal-regulated kinase (p-ERK), phosphorylated c-Jun N-terminal kinase (p-JNK), and phosphorylated P38 were measured by Western blot and/or real-time polymerase chain reaction. Fluorofenidone attenuated TGF-ß1-induced expressions of α-SMA, fibronectin, and collagen I; inhibited phosphorylation of ERK, JNK, and P38; and restored caveolin-1 protein expression but cannot increase caveolin-1 mRNA level in vitro. After caveolin-1 was silenced, FD could not downregulate TGF-ß1-induced expressions of α-SMA, fibronectin, and collagen I or phosphorylation of ERK, JNK, and P38. These studies demonstrate that FD, a potential antifibrotic agent, may attenuate TGF-ß1-induced activation of NHLFs by restoring the expression of caveolin-1.

Draft genome sequence of Brevibacillus brevis strain X23, a biocontrol agent against bacterial wilt.

Brevibacillus brevis X23 is an appropriate biocontrol agent against bacterial wilt caused by Ralstonia solanacearum. We report herein the draft genome sequence (6,566,879 bp) and a circular plasmid (6,600 bp) of B. brevis X23, data which may be helpful for mining the antagonistic activity against R. solanacearum.

[Change in serum levels of soluble Fas and soluble FasL in patients with multiple organ dysfunction syndrome].

OBJECTIVE: To evaluate the significance of changes in the serum levels of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients with multiple organ dysfunction syndrome (MODS) regarding its diagnosis and judgement of severity and outcome, and to investigate the correlations between the levels of tumor necrosis factor-alpha (TNF-alpha), sFas and sFasL. METHODS: Enzyme linked immunoadsorbent assay (ELISA) was used in the determination of serum sFas, sFasL and TNF-alpha in 36 patients with MODS. Thirty-two non SIRS patients and 20 healthy individuals comprised the control groups. The acute physiology and chronic health evaluation III (APACHE III) scoring system and modified MODS scoring system were used to assess patients' clinical severity. The differences of sFas and sFasL levels between MODS group and control groups and between survival and dead patients were observed. The correlations between sFas, sFasL and TNF-alpha levels and severity of MODS and the correlations between the TNF-alpha levels and the levels of sFas and sFasL were also observed. RESULTS: The serum levels of sFas, sFasL and TNF-alpha in patients with MODS were significantly higher than those in controls (P<0.05 or P<0.01), and were associated with severity of the disease (all P<0.01) . The sFas and sFasL levels were found to be significantly higher in the patients who eventually died as compared to those in the patients who survived (both P<0.05). Positive correlations were noted between the TNF-alpha levels and the levels of sFas and sFasL(both P<0.01). The serum levels of sFas and sFasL were elevated with the increase of the number of failure organs in MODS patients. CONCLUSION: The serum levels of sFas and sFasL may serve as diagnostic and prognostic indicators for MODS. TNF-alpha may help enhance the expression of Fas/FasL system.

Molecular diversity of nitrogen-fixing bacteria from the Tibetan Plateau, China.

To understand the composition and structure of nitrogen-fixing bacterial communities from the Sanjiangyuan Nature Reserve on the Tibetan Plateau, the molecular diversity of nifH genes from soil obtained at six sites was examined using a PCR-based cloning approach. Six samples were collected from different regions at an altitude of 3907-4824 m above sea level, and a principal component analysis (PCA) showed that they had different biogeochemical properties. A total of 446 clones and 162 unique RFLP patterns were found. PCA of the RFLP patterns and their biogeochemical parameters showed that the content of soil organic carbon (C), total nitrogen (N) and altitude were the most important factors affecting the nitrogen-fixing bacteria community. Fifty-nine nifH clones were sequenced and their nucleotide identity varied from 64% to 98%, subdivisible into four groups in our phylogenetic tree. Some of the clone sequences were related to nifH genes belonging to four phylogenetic subdivisions (alpha, beta, gamma and delta subclasses of the Proteobacteria), while most of the clones were closely related to the genes of the uncultured bacteria. The tree also showed that the sequence distributions were not clearly related to the sample sites.

[Extraction method of soil microbial DNA for molecular ecology study].

In environmental microbiology, molecular ecology study has been widely concerned in the world, while high quality DNA is the basis of the study. In our study, soil microbial DNA was extracted by the methods of SDS lysis denaturant plus SDS lysis, and purified with gel electrophoresis plus minicolumn and double minicolumn methods. The results showed that denaturant plus SDS lysis could extract DNA more efficiently, and gel electrophoresis plus minicolumn could help to obtain purer DNA that was available in amplifying its 16S rDNA and functional genes by PCR. Therefore, denaturant plus SDS lysis could be an efficient and reliable method to extract DNA molecular ecology studies.

[The community and structure of nitrogen-fixing microorganism in Sanjiangyuan natural reserve].

Research on the diversity of microorganism community in natural environment has been concerned hot spot using the newly molecular biotechnology in the world now. To understand the composition and structure of nitrogen-fixing bacteria communities in the Qingzang plateau, the molecular diversity and phylogenetic of nifH genes of Sangjiangyuan natural reserve were examined by using the PCR-RFLP based cloning approach. The 3 samples were come from different sites and different plant types, and their biogeochemical parameters were diverse. DNA was directly extracted from the soil microorganism and amplified the nifH gene fragment using PCR by the primers of nifH-34F 5'-AAAGG(C/T)GG(A/T) ATCGG(C/T)AA(A/G) TCCACCAC-3' and nifH-491R 5'-TFGTT(G/C)GC(G/C)GC(A/G)TACAT(G/C)GCCATCAT-3'. For the nifH gene segment, diverse PCR products were characterized by cloning, restriction fragment length polymorphism (RFLP) analysis and sequencing. A total of 233 clones and 99 operational taxonomic units (OTUs) which were digested the clones by the restriction enzymes MspI and RsaI were obtained from all samples. YS-1 had 63 clones and 24 OTUs, ZD-1 had 75 clones and 28 OTUs, and NQ-1 had 95 clones and 47 OTUs, respectively. They were found 1-2 significant domain groups of clones and shared 4 OTUs in all samples. A wide range of sequence divergence was observed in the 26 nifH clones that were sequenced from all samples. Sequence comparison showed that the nifH clones were 66% to 98% similar. The phylogenetic tree was constructed by the Clustal W and Mega software. 26 sequences could be subdivided into 4 clusters in the phylogenetic tree, and some of them had the closely similar to Proteobacteria, but The majority of the clones were not closely related to any known cultivated nitrogen-fixing bacteria, Therefore, most of them are unique and may represent novel sequences of nitrogen-fixing bacteria.

[Molecular diversity and phylogenetic analysis of nitrogen-fixing (nifH) genes in alp prairie soil of Sanjiangyuan natural reserve].

Research on the diversity of microorganism community in natural environment has been concerned hot spot using the newly molecular biotechnology in the world now. This was the first description of the molecular diversity and phylogenetic analysis of nitrogen-fixing (nifH) genes in alp prairie soil of Sanjiangyuan natural reserve. DNA was directly extracted from the soil microorganism and amplified the nifH gene fragment using PCR by the primers of nifH-34F 5'-AAAGG(C/T)GG(A/T) ATCGG(C/T)AA(A/G) TCCACCAC-3' and nifH-491R 5'-TYGTT(G/C)GC(G/C)GC(A/G)TACAT(G/C)G CCATCAT-3'. For the gene fragment, diverse PCR products were characterized by cloning, restriction fragment length polymorphism (RFLP) analysis and sequencing. 143 clones and 35 different RFLP patterns were received in two samples by the restriction enzymes MspI and RsaI digested. ZD sample had 82 clones and 21 different RFLP patterns, and YS sample had 61 clones and 19 different RFLP patterns. There were shared 5 RFLP patterns in two samples. The analysis result found a significant dominant group of clones occurring in both samples which account for 29.3% and 32.8%, respectively, and several minor groups were also detected. 21 clones were sequenced, and their levels of nucleotide identity were from 71% to 98%. None of the sequenced nifH gene was completely identical to any deposited in the data banks, and therefore each of them belong to a noncharacterized bacterium. Finally, the phylogenetic tree was constructed by the Clustal W and Mega software. 21 sequences can be subdivided into 4 clusters in the phylogenetic tree, and most of them had the closely similar toalpha- , beta-, and gamma-Proteobacteria . The significant dominant group in YS sample and ZD sample had the closely related with Rhodobacter sphaeroides and Delftia tsuruhatensis, respectively. The YS-nifH-11 was the only sequence which had highly similar to Cyanobacteria .

[Genetic diversity of microsatellite loci in captive Amur tigers].

The tiger is one of the most threatened wildlife species since the abundance and distribution of tiger have decreased dramatically in the last century. The wild Amur tiger (Panthera tigris altaica) only distributed in northeast China, the far east area of Russia and the north Korea and its size of wild population is about 450 in the world and 20 in China. Several hundred captive populations of Amur tigers are the main source to protect gene library of tiger and the source of recovering the wild populations. The Breeding Center for Felidae at Hengdaohezi and Haoerbin Tiger Park in Heilongjiang Province is the biggest captive breeding base in China. How to make clear the genetic pedigree and establish reasonable breeding system is the urgent issues. So we use the microsatellite DNA markers and non-invasive technology to research on the genetic diversity of captive Amur tiger in this study. Ten microsatellite loci (Fca005, Fca075, Fca094, Fca152, Fca161, Fca294, Pti002, Pti003, Pti007 and Pti010), highly variable nuclear markers, were studied their genetic diversity in 113 captive Amur tigers. The PCR amplified products of microsatellite loci were detected by non-denatured polyacrylamide gel electrophoresis. Allele numbers, allelic frequency, gene heterozygosity(H(e)), polymorphism information content(PIC) and effective number of allele(N(e)) were calculated. 41 alleles were found and their size were ranged from 110bp to 250bp in ten microsatellite loci, Fca152 had 6 alleles, Fca075, Fca094 and Fca294 had 5 alleles, Fca005 and Pti002 had 4 alleles and the others had 3 alleles in all tiger samples, respectively. The allelic frequencies were from 0.009 to 0.767; The He ranged from 0.385 to 0.707, and Fca294 and Pti010 locus had the highest and lowest value; the PIC were from 0.353 to 0.658, Fca294 and Pti010 locus had the highest and lowest value; and N(e) were from 1.626 to 3.409, Fca294 and Pti010 locus had the highest and lowest value, which showed the ten microsatellie loci had high or medium polymorphism in these Amur tigers and had high genetic diversity. At the same time, we only found even bases variability which showed the even bases repeat sequence (CA/GT) maybe the basic unit for length variability of microsatellite in all loci. In this study, the samples were made up of 75 hair specimens, 23 blood specimens and 15 tissue specimens, we obtained the genome DNA from hairs using the non-invasive DNA technology and demonstrated that DNA derived from hair samples is as good as that obtained from blood samples for the analysis of microsatellite polymorphism. These results imply that microsatellite DNA markers and non-invasive DNA technology can help study the genetic diversity of Amur tiger. This method could be used in the captive management of other endangered species.