Oxidation of Hypophosphorous Acid by a Ruthenium(VI) Nitrido Complex in Aqueous Acidic Solution. Evidence for a Proton-Coupled N-Atom Transfer Mechanism.

The oxidation of hypophosphorous acid (H3PO2) by a ruthenium(VI) nitrido complex, [(L)RuVI(N)(OH2)]+ (RuVIN; L = N,N'-bis(salicylidene)-o-cyclohexyldiamine dianion), has been studied in aqueous acidic solutions at pH 0-2.50. The reaction has the following stoichiometry: 2[(L)RuVI(N)(OH2)]+ + 3H3PO2 + H2O → 2[(L)RuIII(NH2P(OH)2)(OH2)]+ + H3PO3. The pseudo-first-order rate constant, kobs, depends linearly on [H3PO2], and the second-order rate constant k2 depends on [H+] according to the relationship k2 = k[H+]/([H+] + Ka), where k is the rate constant for the oxidation of H3PO2 molecule and Ka is the dissociation constant of H3PO2. At 298.0 K and I = 1.0 M, k = (2.04 ± 0.19) × 10-2 M-1 s-1 and Ka = (6.38 ± 0.63) × 10-2 M. A kinetic isotope effect (KIE) of 2.9 ± 0.1 was obtained when kinetic studies were carried out with D3PO2 at pH 1.16, suggesting P-H bond cleavage in the rate-determining step. On the other hand, when the kinetics were determined in D2O, an inverse KIE of 0.21 ± 0.03 (H3PO2 in H2O vs H3PO2 in D2O) was found. On the basis of experimental results and DFT calculations, the proposed mechanism involves an acid-catalyzed tautomerization of H2P(O)(OH) to HP(OH)2; the latter molecule is the reacting species which reacts with RuVIN via a proton-coupled N-atom transfer pathway.

In vitro anti-platelet aggregation effects of fourteen fruits and vegetables.

In the present study, the anti-platelet aggregation activity of 14 vegetables and fruits was tested in vitro. The aqueous, 90% ethanol and ethyl acetate extracts, as well as concentrated juices of 14 foods (fruits and vegetables) were prepared, and the anti-platelet aggregation activity of those extracts was analyzed on a platelet aggregation analyzer in vitro with adenosine 5'-diphosphate (ADP), bovine thrombin (THR) and arachidonic acid (AA) as aggregation inducers, respectively. Aspirin (ASP) was used as the positive control. A number of the tested foods had inhibitory effects in concentration-dependent manner on platelet aggregations induced by various agonists. Especially, some foods such as lemon, leek, garlic, scallion, ginger, tomato and grapefruit showed good anti-platelet aggregation effect similar or higher than that of positive control group i.e. aspirin (ASP). The results of present study provide scientific reference for reasonable selection of daily dietary with supplementary curative effects or prevention of cardiovascular diseases (CVD).

Evaluation of interactions between RAW264.7 macrophages and small molecules by capillary electrophoresis.

In this study, the affinity interactions between RAW 264.7 macrophages and three small molecules including naringin, oleuropein and paeoniflorin were evaluated by affinity capillary electrophoresis (ACE), partial filling affinity capillary electrophoresis (PFACE) and frontal analysis capillary electrophoresis (FACE), respectively. The result indicated that ACE (varying concentrations of cell suspension were filled in the capillary as receptor) may not be suitable for the evaluation of interactions between cell and small molecules due to the high viscosity of cell suspension; PFACE can qualitatively evaluate the interaction, but the difference in viscosity between RAW264.7 suspension and buffer effects on the liner relationship between filling length and injection time, which makes the calculation of binding constant difficult. Furthermore, based on the PFACE results, naringin showed stronger interaction with macrophages than the other two molecules; taking advantage of the aggregation phenomenon of cell induced by electric field, FACE was successfully used to determine the stoichiometry (n = 5×109 ) and binding constant (Kb = 1×104 L/mol) of the interaction between RAW264.7 and naringin.

Evaluation of affinity interaction between small molecules and platelets by open tubular affinity capillary electrochromatography.

In this paper, an open tubular affinity capillary electrochromatography (OT-ACEC) was developed by physical adsorption of rabbit platelets on the inner surface of capillary. The interactions between small molecules include adenosine diphosphate (ADP) (positive control), protocatechuic acid (negative control) and seven natural products (salvianolic acid B, salvianic acid A sodium, hydroxysafflor yellow A, ferulic acid, chlorogenic acid, sinapic acid, caffeic acid) and platelets were evaluated by their retention factors and binding constants obtained based on peak-shift assay. Then, the activities of anti-platelet aggregation induced by thrombin (THR), ADP and arachidonic acid (AA) for those small molecules (except ADP) were evaluated by turbidimetric method. The results indicate that: (i) ADP, a platelet aggregation inducer, had strong interaction with platelet, while protocatechuic acid that had no inhibition on platelet aggregation behaved no specific interaction; (ii) there was a positive correlation between the anti-platelet aggregation activities of small molecules and their interactions with platelet, generally those compounds with higher binding constants with platelet exhibited higher activities. Therefore, the OT-ACEC method developed in the present study can be a potential method to evaluate affinity interactions between small molecules and platelets, so as to predict the biological activities such as anti-platelet aggregation for the small molecules.

Simultaneous screening and analysis of antiplatelet aggregation active alkaloids from Rhizoma Corydalis.

CONTEXT: The rising problem of atherosclerosis and ischemic heart disease emphasizes the need to look for new antithrombotic components with effective modes of action. Corydalis yanhusuo (Y.H. Chou & Chun C. Hsu) W.T. Wang ex Z.Y. Su & C.Y. Wu (Papaveraceae) (Rhizoma Corydalis) has been used in the traditional medicines for the treatment of cardiovascular disease. OBJECTIVE: The antiplatelet aggregation compounds in Rhizoma Corydalis were screened to validate its traditional medicinal use. MATERIAL AND METHODS: Total alkaloid extract (TAE) of Rhizoma Corydalis was obtained by refluxing 100 g Rhizoma Corydalis powder with 600 mL 70% ethanol, and purified by acidification (20% HCl) and alkalization (5 M NaOH) process. Potential antiplatelet aggregation compounds in TAE were screened by a method involving platelet bio-specific extraction and HPLC-DAD/LC-MS analysis. Further in vitro antiplatelet aggregation activity confirmation of TAE and seven main alkaloids were achieved by turbidimetry method within 3 h after blood collection from rabbit carotid artery, and all the test drugs were at the concentration range of 25-350 µg/mL. Finally, HPLC-DAD was employed for the quantitative determination of seven main components in TAE. RESULTS: Five alkaloids, identified as glaucine, dehydrocorydaline, canadine, tetrahydrocoptisine and corydaline, can be specifically extracted with platelets. The results indicated that all these five alkaloids can inhibit thrombin-induced platelet aggregation in a low dose (IC50 of glaucine, dehydrocorydaline, canadine, tetrahydrocoptisine and corydaline were 49.057, 34.914, 33.547, 84.261 and 54.164 µg/mL, respectively) as compared to TAE (IC50 = 175.426 µg/mL) and aspirin (IC50 = 300.340 µg/mL), while the unbound compounds (palmatine and tetrahydropalmatine) had a very weak antiplatelet effect (IC50 > 200 µg/mL). DISCUSSION AND CONCLUSION: This study is the first reported work for antiplatelet components screening in Rhizoma Corydalis. Seven compounds were detected and identified by HPLC-DAD/LC-MS, of which five platelet-targeted compounds were discovered.

[Applications of platelets in studies on traditional Chinese medicines promoting blood circulation to remove blood stasis].

Thrombotic diseases in different forms become a great threat to human health. Such anti-platelet aggregation drugs as aspirin and clopidogrel are common drugs in clinic. However, along with the appearance of resistance and side effects of western anti-platelet aggregation drugs, anti-platelet aggregation traditional Chinese medicines promoting blood circulation to remove blood stasis have gradually become an important study orientation. Platelet is one of major participant in thrombosis, and plays an important role as a bioactive material in studies on traditional Chinese medicines promoting blood circulation to remove blood stasis, mainly involving two aspects--the evaluation for the anti-platelet aggregation activity of traditional Chinese medicines and the screening of their active components. This paper summarized the applications of platelets in studies on traditional Chinese medicines promoting blood circulation to remove blood stasis, so as to provide basis for further studies.

Design, Synthesis, and Biological Activity of Donepezil: Aromatic Amine Hybrids as Anti-Alzheimerss Drugs.

In this study, benzylpiperidine, the active group of donepezil (DNP), was connected with the neurotransmitter phenylethylamine by square amide, in which the fat chain of phenylethylamine was reduced and the benzene rings were substituted. A series of multifunctional hybrid compounds, including DNP-aniline hybrids (1-8), DNP-benzylamine hybrids (9-14), and DNP-phenylethylamine hybrids (15-21) were obtained and their cholinesterase inhibitory activity and neuroprotection of the SH-SY5Y cell line were determined. Results showed that compound 3 exhibited excellent acetylcholinesterase inhibitory activity with an IC50 value of 4.4 µM, higher than that of positive control DNP and significant neuroprotective effects against H2O2-induced oxidative damage in SH-SY5Y cells with 80.11% viability rate at 12.5 µM, much higher than that of the model group (viability rate = 53.1%). The mechanism of action of compound 3 was elucidated by molecular docking, reactive oxygen species (ROS), and immunofluorescence analysis. The results suggest that compound 3 could be further explored as a lead compound for the treatment of Alzheimer's disease. In addition, molecular docking research indicated that the square amide group formed strong interactions with the target protein. Based on the above analysis, we believe that square amide could be an interesting construction unit in anti-AD agents.

Clinical Efficacy of Blood Ultrafiltration Therapy in Patients with Acute Decompensated Chronic Heart Failure Running Title: Blood Ultrafiltration Therapy for Heart Failure.

In this study, we investigated the clinical effects of blood ultrafiltration therapy in patients with acute decompensated chronic heart failure. We enrolled 78 patients with acute decompensated chronic heart failure who were admitted to a hospital from September 2017 to December 2021, and divided them into two groups based on the digital randomization method. The FQ-16 heart failure ultrafiltration dehydrating device blood ultrafiltration therapy was administered to the observation group (39 patients) for 8-16 hours, while the control group (39 patients) received the stepped drug therapy. Echocardiography was used to assess the changes in cardiac function of the patients in both groups before and after treatment. The changes in urine volume, N-terminal pro-B-type natriuretic peptide (NT-proBNP), plasma renin, and serum creatinine levels were measured before and after the treatment to compare the overall response rate of the patients in both groups. The differences in left ventricular end-systolic dimension and left ventricular end-diastolic dimension and the ejection fraction between the groups before treatment were not statistically significant (P > 0.05), however, the left ventricular end-diastolic dimension in the observation group was significantly lower and the ejection fraction was significantly higher (P < 0.05) compared with that before treatment; the urine volume, N-terminal pro-B-type natriuretic peptide (NT-proBNP), plasma renin, and serum creatinine were significantly improved in both groups after treatment compared with that before treatment. All indexes in the observation group were better than those in the control group (P < 0.05), 74.36%. The overall response rate of the observation group was 94.87%, x2 = 4.843 and the difference between groups was statistically significant (P < 0.05). Blood ultrafiltration therapy for patients with acute decompensated chronic heart failure can improve their cardiac and renal functions, reduce NT-proBNP, reduce volume load, and enhance efficacy while ensuring high safety.

Mammalian target of rapamycin regulates isoliquiritigenin-induced autophagic and apoptotic cell death in adenoid cystic carcinoma cells.

Previous studies, including those from our laboratory, have demonstrated that isoliquiritigenin (ISL), a flavonoid isolated from licorice, is a promising cancer chemotherapeutic agent. However the mechanisms underlying its anticancer effects are still far from clear. We now show, for the first time, that ISL triggers the mammalian target of rapamycin (mTOR)-dependent autophagic and apoptotic cell death in adenoid cystic carcinoma (ACC). Exposure of both ACC-2 and ACC-M cells to ISL resulted in several specific features for autophagy, including the appearance of membranous vacuoles, formation of acidic vesicular organelles, punctate pattern of LC3 immunostaining, and an increase in autophagic flux. Moreover, ISL treatment also resulted in significantly increased apoptosis in ACC cells. The ISL-mediated autophagic and apoptotic cell death were obviously attenuated by transfection with dominant negative Atg5 (DN-Atg5(K130R)) plasmids or treatment with 3-methyladenine(3-MA). In additon, the data also revealed that the autophagic and apoptotic cell death induced by ISL occurred through a mTOR-dependent pathway. More importantly, the xenograft model using ACC-M cells provided further evidence of the occurrence of ISL-induced autophagy and apoptosis in vivo, correlating with the suppresson of mTOR activation as well as up-regulation of Atg5 expression. Taken together, these findings in our study suggest that induction of mTOR-dependent autophagic and apoptotic cell death may be an important mechanism in cancer chemotherapy by ISL.

Salicylic Acid Enhances Heat Stress Resistance of Pleurotus ostreatus (Jacq.) P. Kumm through Metabolic Rearrangement.

Pleurotus ostreatus (Jacq.) P. Kumm is cultivated worldwide, and its growth is seriously threatened by heat stress. Here, we performed a comprehensive analysis to investigate the influence of the phytohormone salicylic acid (SA) in P. ostreatus under HS. The results showed that the hyphal growth recovery rate and the antioxidant capacity of P. ostreatus increased with exogenous SA application (0.01 mmol/L and 0.05 mmol/L) after HS treatment. Metabolomic and transcriptomic analyses showed that SA application (0.05 mmol/L) weakened central carbon metabolism to allow cells to survive HS efficiently. In addition, SA shifted glycolysis to one-carbon metabolism to produce ROS scavengers (GSH and NADPH) and reduced ROS production by altering mitochondrial metabolism. SA also maintained nucleotide homeostasis, led to membrane lipid remodeling, activated the MAPK pathway, and promoted the synthesis of cell-wall components. This study provides a reference for further study of SA in microorganisms.

Elevated Sad1 and UNC84 Domain Containing 2 (SUN2) level inhibits cell growth and aerobic glycolysis in oral cancer through reducing the expressions of glucose transporter 1 (GLUT1) and lactate dehydrogenase A (LDHA).

BACKGROUND/PURPOSE: Oral cancer is a malignant tumor accompanied by high morbidity, mortality, and poor prognosis. Therefore, it is urgent to explore the percise regulation mechanisms underlying oral cancer. Sad1 and UNC84 Domain Containing 2 (SUN2) was considered as a tumor suppressor in some cancers. The purpose of the study was to define the role of SUN2 in oral cancer progression. MATERIALS AND METHODS: Tumor tissues and paired paracancerous healthy tissues from 56 oral cancer patients were collected. Cell viability was measured using MTT assay. The colony formation assay was applied to determine cell proliferation ability. The mRNA and protein levels were assessed by qRT-PCR and Western blot, respectively. RESULTS: SUN2 expression was decreased in oral cancer tissues and cell models. SUN2 overexpression suppressed the growth of oral cancer cells, while the down-regulation of SUN2 promoted cell growth. SUN2 overexpression restrained the glucose uptake, lactate production, and ATP level of oral cancer cells, whereas down-regulation of SUN2 promoted glycolysis. Besides, elevated SUN2 inhibited the glucose transporter 1 (GLUT1) and lactate dehydrogenase A (LDHA) levels. However, SUN2 knockdown increased the levels of GLUT1 and LDHA. CONCLUSION: SUN2 was decreased in oral cancer in vivo and in vitro. SUN2 overexpression suppressed cell growth and glycolysis via reducing the levels of GLUT1 and LDHA in oral cancer.

Poly[[aqua-µ(3)-2,2-dimethylmalonato-copper(II)] monohydrate] and poly[aqua-µ(3)-2,2-dimethylmalonato-copper(II)].

The coordination mode of the dimethylmalonate ligand in the two title Cu(II) complexes, {[Cu(C(5)H(3)O(4))(H(2)O)]·H(2)O}(n), (I), and [Cu(C(5)H(3)O(4))(H(2)O)](n), (II), is the same, with chelated six-membered, bis-monodentate and bridging bonding modes. However, the coordination environment of the Cu(II) atoms, the connectivity of their metal-organic frameworks and their hydrogen-bonding interactions are different. Complex (I) has a perfect square-pyramidal Cu(II) environment with the aqua ligand in the apical position, and only one type of square grid consisting of Cu(II) atoms linked via carboxylate bridges to three dimethylmalonate ligands, with weak hydrogen-bond interactions within and between its two-dimensional layers. Complex (II) has a coordination geometry that is closer to square pyramidal than trigonal bipyramidal for its Cu(II) atoms with the aqua ligand now in the basal plane. Its two-dimensional layer structure comprises two alternating grids, which involve two and four different dimethylmalonate anions, respectively. There are strong hydrogen bonds only within its layers.

Poly[diaquabis(mu3-2,2-dimethylpropanedioato)calcium(II)copper(II)].

The title complex, [CaCu(C(5)H(6)O(4))(2)(H(2)O)(2)](n), is the first heterobimetallic complex based on a substituted malonate dianion. The Cu(II) cation and two independent 2,2-dimethylmalonate (or 2,2-dimethylpropanedioate) dianions build up a robust dianionic [Cu(C(5)H(6)O(4))(2)](2-) complex, which acts as a building block to coordinate to four Ca(2+) cations. Each Cu(II) centre is in a four-coordinate square plane of dimethylmalonate O atoms, while each Ca(II) atom is in an eight-coordinate distorted bicapped trigonal-prismatic environment of six O atoms from four different dimethylmalonate groups and two water molecules. This arrangement creates a two-dimensional layer connectivity of the structure. The dianionic [Cu(C(5)H(6)O(4))(2)](2-) units are involved in different intermolecular hydrogen-bonding interactions with water molecules via the formation of hydrogen-bonded rings of graph sets R(1)(2)(8) and R(6) within this layer. The crystal was nonmerohedrally twinned by rotation about [011] with a major twin volume fraction of 0.513 (3).

4,4'-Bipyridine-3-nitro-benzoic acid (1/2).

The title compound, C(10)H(8)N(2)·2C(7)H(5)NO(4),was obtained unintentionally as the harvested product of the hydro-thermal reaction between Co(OAc)(2)·4H(2)O and 4,4'-bipyridine in the presence of 3-nitro-phthalic acid. In the reaction, 3-nitro-phthalic acid is transformed into 3-nitro-benzoic acid by an in situ deca-rboxylation reaction, in which the carboxyl-ate group is not deprotonated and is uncoordinated. In the crystal, the uncoordinated 3-nitro-benzoic acid and free 4,4'-bipyridine mol-ecules are linked alternately by O-H⋯N hydrogen bonds into chains, which are assembled by C-H⋯O hydrogen bonds into a three-dimensional supra-molecular network.

Poly[[di-mu-aqua-tetraaquadi-mu-hydroxido-bis(mu3-3-nitrophthalato)tricopper(II)] dihydrate].

The novel title complex, {[Cu(3)(C(8)H(3)NO(6))(2)(OH)(2)(H(2)O)(6)].2H(2)O}(n), has a one-dimensional polymeric double chain structure where the three Cu atoms are linked by mu(2)-OH and mu(2)-H(2)O groups, and these trinuclear centres are bridged by two 3-nitrophthalate ligands. The asymmetric unit contains one and a half crystallographically independent Cu atoms (one lying on a centre of inversion), both coordinated by six O atoms and exhibiting distorted octahedral coordination geometries, but with different coordination environments. Each 3-nitrophthalate ligand connects to three Cu atoms through two O atoms of one carboxylate group and one O atom of the nitro group. The remaining carboxylate group is free and is involved in intrachain hydrogen bonds, reinforcing the chain linkage.

Bradyrhizobium elkanii, Bradyrhizobium yuanmingense and Bradyrhizobium japonicum are the main rhizobia associated with Vigna unguiculata and Vigna radiata in the subtropical region of China.

Cowpea (Vigna unguiculata) and mung bean (Vigna radiata) are important legume crops yet their rhizobia have not been well characterized. In the present study, 62 rhizobial strains isolated from the root nodules of these plants grown in the subtropical region of China were analyzed via a polyphasic approach. The results showed that 90% of the analyzed strains belonged to or were related to Bradyrhizobium japonicum, Bradyrhizobium liaoningense, Bradyrhizobium yuanmingense and Bradyrhizobium elkanii, while the remaining represented Rhizobium leguminosarum, Rhizobium etli and Sinorhizobium fredii. Diverse nifH and nodC genes were found in these strains and their symbiotic genes were mainly coevolved with the housekeeping genes, indicating that the symbiotic genes were mainly maintained by vertical transfer in the studied rhizobial populations.

Hexaaquanickel(II) bis[2-carboxylato-2-(isothiouronium-S-ylmethyl)propane-1,3-diyl phosphate] hexahydrate.

In the title complex, [Ni(H(2)O)(6)](C(6)H(10)N(2)O(6)PS)(2) x 6 H(2)O, the asymmetric unit consists of one-half of an Ni atom (which lies on an inversion centre) with three coordinated water molecules, one complete 2-carboxylato-2-(isothiouronium-S-ylmethyl)propane-1,3-diyl phosphate anion and three noncoordinated water molecules. The hexaaquanickel(II) cations have distorted octahedral coordination and are connected via water chains to form two-dimensional supramolecular networks parallel to the ab plane. The phosphate ester anion is linked via N-H...O and O-H...O hydrogen bonds, thus creating various ring, dimer and chain hydrogen-bonding patterns, and building up a second two-dimensional supramolecular network parallel to the ab plane. The crystal structure is further stabilized by an intra- and interlayer hydrogen-bond network. This work illustrates that a carboxylate with a caged phosphate ester can open its ring in the presence of dichloridotetrakis(thiourea)nickel, and the resulting polyfunctional anion can be used for constructing a complex hydrogen-bonding scheme.

Aqua-(2,2'-bipyrid-yl)(pyrazine-2,6-dicarboxyl-ato)nickel(II) 1.25-hydrate.

The asymmetric unit of the title compound, [Ni(C(6)H(2)N(2)O(4))(C(10)H(8)N(2))(H(2)O)]·1.25H(2)O, contains two independent chemically identical Ni(II) complex cations and two and a half solvent water mol-ecules. The Ni(II) ions are in slightly distorted coordination environments. In the crystal structure, inter-molecular O-H⋯O and weak C-H⋯O hydrogen bonds link cations and water mol-ecules into a three-dimensional network. One of the three uncoordinated water molecules is half-occupied.

Prognostic relevance of tripartite motif containing 24 expression in colorectal cancer.

Colorectal cancer is one of the most frequent malignancies in the world. Tripartite motif containing 24 (TRIM24) is a member of the TRIM protein family and a coregulator for multiple nuclear receptors. Altered expression of TRIM24 has been shown in a spectrum of human cancers. However, the clinical role of TRIM24 in colorectal cancer remains unknown. Here, gene expression data in colorectal cancer and normal tissues were downloaded from Gene Expression Omnibus (GEO). Western blotting analysis was conducted to compare TRIM24 expression between colorectal cancer and non-cancerous tissues. Immunohistochemistry staining were performed to assess TRIM24 expression in colorectal cancer tissues, and statistical analyses were employed to evaluate the associations of TRIM24 expression with clinicopathologic features and overall survival. TRIM24 mRNA and protein levels were higher in colorectal cancer tissues than that in the normal controls. TRIM24 protein expression was positively correlated with tumor size (P=0.0269), clinical stage (P=0.0061), vital status (P=0.0110) and serum carcinoembryonic antigen levels (P=0.0176). Kaplan-Meier survival analysis indicated that patients with higher TRIM24 expression had shorter survival time than those with lower TRIM24 expression. Multivariate analyses revealed TRIM24 expression was an independent prognostic factor (P<0.001). In conclusion, our study suggests that TRIM24 may play a role in colorectal carcinogens and serve as a potential prognostic marker of human colorectal cancer.

Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis.

BACKGROUND: As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. OBJECTIVE: Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. MATERIALS AND METHODS: Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). RESULTS: The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. CONCLUSIONS: Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. SUMMARY: The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis: CS, TCMs: Traditional Chinese medicines.