Molecular Mechanism of Electroacupuncture Regulating Cerebral Arterial Contractile Protein in Rats with Cerebral Infarction Based on MLCK Pathway.

OBJECTIVE: To explore the effect of electroacupuncture (EA) intervention on the vasoconstriction of cerebral artery smooth muscle cells after cerebral infarction. METHODS: Male Wistar rats were randomly divided into 3 groups by a random number table: the model group (n=24), the EA group (n=24), and the normal group (n=6). The model and the EA groups were divided into different time subgroups at 0.5, 1, 3, and 6 h after middle cerebral artery occlusion (MCAO), with 6 rats in each subgroup. MCAO model was established using intraluminal suture occlusion method. The EA group was given EA treatment at acupoint Shuigou (GV 26) instantly after MCAO for 20 min. The contents of cerebrovascular smooth muscle MLCK, the 3 subunits of myosin light chain phosphatase (MLCP) MYPT1, PP1c-δ and M20, as well as myosin-ATPase activity were detected using immunohistochemistry and Western blotting. RESULTS: The overall expression level of the MYPT1 and PP1c-δ in the model group was significantly higher (P<0.01). After EA intervention, the 0.5 h group expression level was close to that of the normal group (P>0.05), and the other subgroups were still significantly higher than the normal group (P<0.01). After EA intervention, the expression level of each subgroup was significantly lower than the corresponding model group. There was a significant difference between the 0.5 and 1 h subgroups (P<0.01), while a difference was also observed between the 3 and 6 h subgroups (P<0.05). The dynamic change rule gradually increased with the prolongation of infarction time within 6 h after infarction. CONCLUSION: EA intervention can inhibit contraction of cerebral vascular smooth muscle cells and regulate smooth muscle relaxation by regulating MLCK pathway.

[Acupuncture for improving neuroinflammation in vascular dementia rats by regulating Th1/Th2 cells balance in peripheral blood].

OBJECTIVE: To observe the effect of acupuncture on the balance of T helper (Th) 1/Th2 cells in peripheral blood, inflammatory reaction and intracerebral neuroinflammation in vascular dementia (VD) rats, and to explore the mechanism of acupuncture for improving cognitive function in VD. METHODS: A total of 60 SPF Wistar rats were randomized into a normal group (n=12), a sham operation group (n=12) and an operation group (n=36). Bilateral common carotid artery occlusion was adopted to establish the VD model in rats of the operation group. The rats of successful modeling were randomized into a model group and an acupuncture group, 12 rats in each one. In the acupuncture group, Sanjiao acupuncture was applied at "Danzhong" (CV 17), "Zhongwan" (CV 12), "Qihai" (CV 6), "Xuehai" (SP 10) and "Zusanli" (ST 36), the needles were manipulated for 30 s at each acupoint, without retaining. The intervention was given once a day for 15 days, and there was 1-day rest on day 8. Morris water maze test was adopted to observe the ethology, flow cytometry was used to detect the ratio of Th1/Th2 in peripheral blood, and Luminex liquid chip technology was used to detect the levels of interleukin (IL)-4, IL-10, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in serum and hippocampus. RESULTS: There were no significant differences in various indexes between the normal group and the sham operation group (P>0.05). Compared with the sham operation group, in the model group, the escape latency of hidden platform test and reversal platform test was prolonged (P<0.01), the residence time of the original platform quadrant was shortened and the number of crossing the original platform was reduced in probe test (P<0.01, P<0.05), the proportion of Th1 cells was increased, the proportion of Th2 cells was decreased and the ratio of Th1/Th2 cells was increased in peripheral blood (P<0.01), the levels of TNF-α and IFN-γ were increased, the levels of IL-4 and IL-10 were decreased in serum and hippocampus (P<0.05, P<0.01). Compared with the model group, in the acupuncture group, the escape latency of hidden platform test and reversal platform test was shortened (P<0.01), the residence time of the original platform quadrant of the probe test was prolonged (P<0.05), the proportion of Th1 cells was decreased, the proportion of Th2 cells was increased and the ratio of Th1 / Th2 cells was decreased in peripheral blood (P<0.05), the levels of TNF-α and IFN-γ were decreased, the levels of IL-4 and IL-10 were increased in serum and hippocampus (P<0.05, P<0.01). CONCLUSION: Acupuncture can improve the cognitive dysfunction and reduce the intracerebral neuroinflammation in VD rats, its mechanism may relate to the regulation of Th1/Th2 cells balance and reduce the inflammatory reaction in peripheral blood.

Effect of Electro-acupuncture on Vasomotor Symptoms in Rats with Acute Cerebral Infarction Based on Phosphatidylinositol System.

OBJECTIVE: To investigate the effect of electro-acupuncture (EA) on vasomotor symptoms in rats with acute cerebral infarction, by observing the changes in the expression of factors related to the phosphatidylinositol (PI) system. METHODS: Forty-two Wistar rats were randomly divided into 3 groups by a random number table: the control group (n=6), the model group (n=18) and the EA group (n=18). The EA group was given EA treatment at Shuigou (GV 26) instantly after modeling with middle cerebral artery occlusion (MCAO) method, while the model and control groups were not given any treatment. The degrees of neurological deficiency were evaluated using neurological severity scores (NSS) and the brain blood flow was evaluated by a laser scanning confocal microscope. Western blot analysis was conducted to detect the expression levels of G-protein subtype (Gq) and calmodulin (CaM). Competition for protein binding was conducted to detect the expression level of inositol triphosphate (IP3). Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol (DAG). The expression level of intracellular concentration of free calcium ion ([Ca2+]i) was detected by flow cytometry. RESULTS: The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO (P<0.01), while the EA group was significantly lower than the model group at 6 h (P<0.01). The cerebral blood flow in the model group was significantly lower than the control group at 1, 3 and 6 h after MCAO (P<0.01), while for the EA group it was remarkably higher than the model group at the same time points (P<0.01). The expressions of Gq, CaM, IP3, DAG and [Ca2+]i in the model group were significantly higher than the control group (P<0.05 or P<0.01), and those in the EA group were significantly lower than the model group at the same time points (P<0.05 or P<0.01). CONCLUSION: EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction, improve cerebral autonomy movement, and alleviate cerebral vascular spasm.

[Using bioinformatics tools to explore cellular biological mechanisms of "Triple Energizer Acupuncture Method" in treating senile dementia].

OBJECTIVE: To observe the effect of needling "Danzhong" (CV17), "Zhongwan"(CV12), "Qihai" (CV6), "Xuehai"(SP10)and "Zusanli"(ST36) (Triple Energizer Acupuncture Method) on the learning-memory ability and profile of hippocampal differentially-expressed genes and proteins of SAMP8 mice (rapid aging mice), so as to explore its underlying mechanisms in improving Alzheimer's disease (AD)．. METHODS: A total of 60 SAMP8 were used as the dementia model and randomly divided into blank control, acupoint and non-acupoint groups (n＝20 per group). The 5 acupuncture points and non-acupoints (subcostal region) on the bilateral sides were needled with filiform needles and manipulated manually for 30 s at each one，once daily, 6 times a week for 4 weeks. The Morris water maze tests (location navigation tests and space probing trials) were used to evaluate the mouse's learning-memory ability. The hippocampal tissue was extracted to detect differentially expressed genes and proteins related to acupuncture intervention by gene chip and isobaric tags for relative and absolute quantitation (iTRAQ) techniques as well as bioinformatic analysis, separately. The information is analyzed through bioinformatics database tools. Finally, immunofluorescence staining was used to verify theresults of microarray analysis. RESULTS: Compared with mice of the control and non-acupoint groups, the escape latency of location navigation task of Morris water maze tests on 4th and 5th day of training was significantly shortened in mice of the acupoint group, and the duration of stay in the original safe-platform quadrant was significantly increased in the acupoint group (P<0.05). Gene microarray displayed that in comparison with the control group, 898 differentially expressed genes were up-regulated, 418 genes were down-regulated in the hippocampus of acupoint group. The iTRAQ analysis indicated that in the acupoint and non-acupoint groups, 286 and 299 differentially expressed proteins were up-regulated, 319 and 179 proteins down-regulated, respectively. Of the 34 terms containing 47 proteins up-regulated by acupoint needling, including intermediate filament, keratin filament, myelin sheath, postsynaptic density, neuron projection were related with neurite and cytoskele-ton. While in the non-acupoint group, of the 24 terms were listed by the system, only the myelin sheath involving 11 differentially expressed proteins functions in activities of neurite and cytoskeleton. Immunofluorescence staining of the hippocampal tissue showed that the high-density distribution areas of neurons and neurite fibers were characterized by decentralization and disordering, with the highlighted areas being mainly near the cell body parts in control mice, but in mice of the acupoint group, the highlighted areas at the neurite were relatively dense, the morphology of hippocampal cells was complete, the fiber structure was clear, dense and orderly, and the neurites were closely arranged and in order, indicating an improvement of the distribution and arrangement of nerve fibers after acupuncture. The height of neurite highlight area of the acupoint group was significantly higher than that of the non-acupoint group (P<0.05)．. CONCLUSION: The "Triple Energizer Acupuncture" of acupoints is able to improve the learning-memory ability in SAMP8, which may be related to its effects in regulating the expression and function of hippocampal genes and proteins related to neurite and cytoskeleton.

Pulmonary Papillary Adenoma: Report of Two Cases.

Pulmonary papillary adenoma is a rare tumor. Two cases without any clinical symptoms were enrolled in our hospital. Both cases were incidentally detected in pulmonary area by imaging. Pathological examination revealed well-circumscribed nodules consisting of papillary growth of cuboidal to low columnar epithelial cells lining the surface of the fibrovascular stroma. Immunohistochemistry (IHC) staining showed that the lining cells were diffusely positive for TTF-1, CK, p63, CK7, and Napsin A. The Ki-67 proliferation index was approximately 2%. The morphological features and the IHC profile of the tumor were in agreement with that of pulmonary papillary adenoma. Both patients are doing well without recurrence or metastasis of the tumor.

[Acupuncture Regulated the Expressions of Hsp84 and Hsp86 and Postponed Brain Aging in SAMP8].

Objective To observe the correlations between heat shock protein 84 ( Hsp84)/ Hsp86 and brain aging in senescence accelerated mouse prone 8 (SAMP8) and the regulation effects of acupuncture. Methods Ten senescence accelerated mouse resistant 1 (SAMR1) were recruited as a normal control group. Another 30 SAMP8 mice were divided into the blank control group, the acupuncture group, and the non-acupoint group by random digit table, 10 in each group. Mice in the acupuncture group received treatment with "Sanjiao" acupuncture method. Mice in the non-acupoint group were needled at two fixed non-acupoints located at bilateral hypochondrium of the body. Catching stimulus at equal volume was given to mice in the rest two groups. All intervention was performed once per day for 15 successive days. Neuromuscular coordination of mice was evaluated. Levels of oxidative stress and protein carbonyl were determined. mRNA and protein expression levels of Hsp84 and Hsp86 in the hippocampus of mice and Neuro-2a cells were detected using Real-time PCR and Western blot. Results Compared with the normal control group, the success rate of tight rope experiment was lowered (P <0. 01) , levels of super- oxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the hippocampus were reduced (P < 0. 01) , the levels of superoxide anion and protein carbonyl increased (P <0. 01, P <0. 05), mRNA and protein expression levels of Hsp84 and Hsp86 in the hippocampus decreased (P <0. 01) in the blank con- trol group. Compared with the blank control group and the non-acupoint group, the success rate of tight rope experiment was elevated (P <0. 05) , levels of SOD and GSH-Px were increased (P <0. 05, P < 0. 01) , the levels of superoxide anion and protein carbonyl decreased (P <0. 01 , P <0. 05), and expres- sion levels of Hsp84 and Hsp86 in the hippocampus increased (P <0. 01) in the acupuncture group. mR- NA expression levels of Hsp84 and Hsp86 were decreased in Neuro-2a cells after treated with AP21-35 (P <0. 01). Conclusions Increased oxidative damage of protein and decreased expression levels of Hsp84 and Hsp86 might be partial reasons for resulting in accumulation of denatured protein and brain aging in SAMP8. Acupuncture could delay brain aging by regulating the expressions of Hsp84 and Hsp86.

[Effect of acupuncture on transmembrane signal pathway in AD mice: an analysis based on lipid-raft proteomics].

OBJECTIVE: To reveal the transmembrane signal pathway participating in regulating neuron functions of treating Alzheimer's disease (AD) by acupuncture. METHODS: SAMP8 mice was used for AD animal model. The effect of acupuncture method for qi benefiting, blood regulating, health supporting, and root strengthening on the amount and varieties of transmembrane signal proteins from hippocampal lipid rafts in SAMP8 mice was detected using HPLC MS/MS proteomics method. RESULTS: Compared with the control group, acupuncture increased 39 transmembrane signal proteins from hippocampal lipid rafts in SAMP8 mice, of them, 14 belonged to ionophorous protein, 8 to G protein, 8 to transmembrane signal receptor, and 9 to kinase protein. Totally 3 main cell signal pathways were involved, including G-protein-coupled receptors signal, enzyme linked receptor signal, and ion-channel mediated signal. Compared with the sham-acupuncture group, acupuncture resulted in significant increase of kinase signal protein amount. From the aspect of functions, they were dominant in regulating synapse functions relevant to cytoskeleton and secreting neurotransmitters. CONCLUSION: The cell biological mechanism for treating AD by acupuncture might be achieved by improving synapse functions and promoting the secretion of neurotransmitters through transmembrane signal transduction, thus improving cognitive function of AD patients.

[Effect of acupuncture intervention on hippocampal neuron loss and astrocytosis in SAMP 8 mice].

OBJECTIVE: To observe the effect of "Sanjiao Acupuncture Therapy" on hippocampal selective neuron loss and astrocytosis in senile accelerated mice (SAM) so as to reveal its mechanism underlying improvement of Alzheimer's disease (AD). METHODS: A total of 18 seven-month old male SAM prone 8 (SAMP 8) were selected and randomly allocated into model group, acupuncture group, and non-acupoint group. Moreover, 6 age- and sex-matched SAM resistant 1 (SAMR 1) were enrolled as the homology normal control group. For mice of the acupuncture group, "Sanjiao Acupuncture Therapy" [manual acupuncture stimulation of "Danzhong" (CV 17), "Zhongwan" (CV 12), "Qihai" (CV 6), "Zusanli" (ST 36) and "Xuehai"(SP 10)] was given, and for rats of the non-acupoint group, manual acupuncture stimulation was applied to bilateral costal region (non-acupoints, below the costal region and 10 and 15 mm superior to the iliac crest). The treatment was conducted once daily for 14 days. At the end of each experiment, the mouse's hippocampus tissue was taken out after routine perfusion and fixing for paraffin sections which were stained by toluidine blue technique for observing changes of neurons and by immunohistochemistry for analyzing glial fibrillary acidic protein (GFAP) immunoactivity of astrocytes. RESULTS: Compared with the control group, a marked neuron loss was found in the model, acupuncture and non-acupoint groups (P < 0.01). The numbers of neurons of hippocampal CA 1 and CA 3 regions in the acupuncture group were significantly more than those in the model group (P < 0.05). Compared with the control group, the expression level of hippocampal GFAP was markedly increased in the model group (P < 0.01). In comparison with the control group, the average expression area, optical density values and the number of GFAP-labeled astocytes were significantly upregulated in the model group (P < 0.01). While in comparison with the model group, these 3 indexes were apparently downregulated in the acupuncture group (P < 0.01) but not in the non-acupoint group except the average expression area, suggesting a reduction of hippocampal astrocyte activity after acupuncture intervention. A negative correlation was found between the expression level of GFAP and the number of astrocytes. CONCLUSION: "Sanjiao Acupuncture Therapy" can lower or delay neuron loss, suppress astrocytosis, and regulate the relationship between neuron and astrocyte, which may contribute to its function in relieving AD.

[Effect of acupuncture on the p53 protein expression of mice with Alzheimer's disease].

OBJECTIVE: To observe the effect of acupuncture method for benefiting qi, regulating blood, supplementing the root, and cultivating the essence (BQRBSRCE) on the p53 protein expression of mice with Alzheimer's disease (AD). METHODS: SAMP8 mice were divided into the control group, the acupuncture group, and the non-acupoint group. The homologous SAMR1 control group was set up. Mice in the acupuncture group used acupuncture method for BQRBSRCE by needling at Tanzhong (RN17), Zhongwan (RN12), Qihai (RN6), and bilateral Xuehai (SP10), and bilateral Zusanli (ST36).Two fixed non-acupoints from bilateral ribs were needled in the non-acupoint group. The p53 protein expression in the cortex and hippocampus of mice was determined using Western blot. The pathological changes of neurons in the hippocampal CA1 region, the temporal lobe, and the occipital lobe were observed using HE staining. The expression of cortical p53-positive cells was detected by immunohistochemical assay. RESULTS: The p53 protein was highly expressed in the cortex of SAMP8, which was significantly down-regulated after acupuncture, showing statistical difference when compared with the SAMP8 control group (P < 0.05), but with no statistical difference when compared with the SAMR1 control group (P > 0.05). Needling at non-acupoints had no obvious effect on the p53 protein expression. There was no statistical difference in the p53 protein expression of the hippocampus (P > 0.05). CONCLUSION: Acupuncture method for BQRBSRCE could down-regulate the p53 protein expression in the brain of mice, improve the pathological state of brain cells, thus enhancing learning and memory capabilities of AD mice, improving their cognitive functions, with specificity of acupoints.

Application of metagenomic techniques in mining enzymes from microbial communities for biofuel synthesis.

Feedstock for biofuel synthesis is transitioning to lignocelluosic biomass to address criticism over competition between first generation biofuels and food production. As microbial catalysis is increasingly applied for the conversion of biomass to biofuels, increased import has been placed on the development of novel enzymes. With revolutionary advances in sequencer technology and metagenomic sequencing, mining enzymes from microbial communities for biofuel synthesis is becoming more and more practical. The present article highlights the latest research progress on the special characteristics of metagenomic sequencing, which has been a powerful tool for new enzyme discovery and gene functional analysis in the biomass energy field. Critical enzymes recently developed for the pretreatment and conversion of lignocellulosic materials are evaluated with respect to their activity and stability, with additional explorations into xylanase, laccase, amylase, chitinase, and lipolytic biocatalysts for other biomass feedstocks.

[Combination of acupoints based on the theory of qi tonification, blood regulation and strengthening the primary source].

The acupuncture technique of tonifying qi, regulating blood and strengthening the primary source of human life is established on the base of the new pathology of dementia due to dysfunction of sanjiao by Prof. HAN Jing-xian. It is held that Tanzhong (CV 17), Zhongwan (CV 12), Qihai (CV 6), Xuehai (SP 10), Zusanli(ST 36) and Waiguan (TE 5) were the basic points for treatment of dementia. There are 3 characteristics of the point combination. Firstly, emphasis should be put on regulation of the stomach and the spleen so as to promote transportation, transformation and distribution; secondly, monarch, minister, assistant and guide points should be selected on the base of meridians, and more attention should be paid to the entity and connections among organs; and thirdly, treatment should be given according to physiological functions of the viscera.

[Effect of puncturing shenshu point on the femoral biomechanical properties in senescence accelerated mouse prone 6].

OBJECTIVE: To explore the mechanisms of puncturing Shenshu point in improving osteoporosis. METHODS: Serum levels of testosterone (T) and osteocalcin (BGP) in senescence accelerated mouse prone 6 (SAMP6, test animals) and senescence accelerated mouse resistant 1 (SAMR1, for control) were determined by radioimmunoassay and their femoral biomechanical properties were determined with three-point bending test before and after puncturing to observe the effect of puncturing on the femoral biomechanical properties and bone mineral contents. RESULTS: Compared with the SAMR1 control group, the serum level of T (20.91 +/- 3.41 nmol/L) decreased (11.09 +/- 1.48 nmol/L in SAMP6 mouse), BGP (6.7 +/- 2.07 microg/L) increased (12.29 +/- 2.29 microg/L in SAMP6 mouse), femoral bending strength lowered and fragility increased. These changes were all improved to some extent or normalized, serum T level 15.05 +/- 2.63 nmol/L and BGP 8.88 +/- 1.85 microg/L after needling at Shenshu point showed significant difference when compared with those in SAMP6. CONCLUSION: Puncturing Shenshu point could effectively prevent the bone loss in SAMP6 mice, increase their bone strength, the therapeutic effect is partly by way of promoting the secretion of sex hormone, improving bone metabolism, suppressing bone transformation rate and increasing bone minerals.

Spatial memory performance and hippocampal neuron number in osteoporotic SAMP6 mice.

The senescence-accelerated mouse strain P6 (SAMP6) is an inbred mouse that represents a clinically relevant model of senile osteoporosis. However, whether osteoporotic SAMP6 mice have cognitive deficits remains largely unexplored. Here, we used Morris water maze to assess reference memory and working memory performance in SAMP6 mice and SAMR1 controls, at 4 and 8 months of age. In addition, unbiased stereological techniques were used to estimate total neuron number in hippocampal CA1 subfield of the mice used in the behavioral study. Morris water maze test revealed impairments in working memory but not in reference memory of the 4- and 8-month-old SAMP6 mice compared with the SAMR1 mice at the same age. However, there were no significant differences in the total numbers of neurons in hippocampal CA1 subfield when comparing 4-month-old SAMR1 and 4-month-old SAMP6 and 8-month-old SAMR1 and 8-month-old SAMP6, which indicate that, in SAMP6 mice, the structural correlates of working memory deficits are to be found in parameters other than the number of neurons in hippocampal CA1 subfield. These findings suggest that SAMP6 mice exhibit selective cognitive deficits and highlight the importance of this mouse model for studying the brain alterations associated with osteoporosis.

Acupuncture prevents cognitive deficits and oxidative stress in cerebral multi-infarction rats.

The aim of this study is to investigate the effects of acupuncture on cognitive deficits and oxidative stress in cerebral multi-infarction rats. The results showed that acupunctural treatment attenuated memory impairment induced by cerebral multi-infarction, as evaluated by shortened escape latency and increased swimming time of rats with memory impairment in the target quadrant. The data additionally suggested that acupunctural treatment ameliorated oxidative injuries induced by cerebral multi-infarction by increasing the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the hippocampus. Further investigation by in situ hybridization and immunohistochemistry revealed that acupunctural treatment significantly increased the expression of CuZnSOD mRNA and protein in the hippocampus of the impaired rats. The findings demonstrate that acupuncture can exert beneficial effects on spatial memory and antioxidant status of cerebral multi-infarction rats.

Gas-initiation under UV and liquid-grafting polymerization on the surface of polysulfone hollow fiber ultrafiltration membrane by dynamic method.

Using the inner-surface of polysulfone hollow fiber ultrafiltration membranes as grafted layer, the method of gas-initiation and liquid-polymerization has been studied, which aimed to adjust the diameter of the pores in the membranes. The degree of polymerization varied with the changes of the parameters, such as irradiation time, monomer concentration, temperature and time of polymerization and soon. The results indicated that using benzophenone (BP) which is in a gaseous condition as photo-initiator, acrylamide as graft monomer, the polyacrylamide chain was grafted on the surface of membranes. After the surface membrane being modified, the water flux and retention altered, and thus it can be seen that the diameter of the pores in the membrane was altered. These experiments contribute to finding a new way to produce the hollow fiber membrane with the small pore size and are extraordinarily worth developing and studying.

Determination of residual clenbuterol in pork meat and liver by HPLC with electrochemical detection.

AIM: To detect the residual clenbuterol in pork meat and liver using HPLC with Coulometric electrode array system. METHODS: Homogenized meat or liver sample was treated with 1 mol x L(-1) hydrochloric acid and centrifuged, the fat existing in meat or liver tissue was removed by diethyl ether. The pH of the remaining aqueous layer was adjusted to 10.8 +/- 0.2 or 11.6 +/- 0.2 for meat or liver and liquid-liquid extraction with diethyl ether was followed. The ether extract was evaporated to dryness, the residue was dissolved in the mobile phase. The mobile phase A consisted of 50 mmol x L(-1) phosphoric acid-30 mmol x L(-1) triethylamine and was adjusted to pH 4.0 with 2 mol x L(-1) sodium hydroxide solution. The mobile phase B consisted of methanol-acetonitrile (30:45). A mixture of mobile phase A and B (80:20) was used in the method. A four electrode array module was selected for quantitation, the electrode potentials were set at 450, 600, 650 and 680 mV respectively. RESULTS: The two calibration curves for meat and liver showed good linearity between 1.88 - 60.16 ng x g(-1), the detection limit of clenbuterol was 1.2 ng x g(-1). CONCLUSION: This method using HPLC-electrochemical detection is reproducible, and the sensitivity is good enough for the determination of clenbuterol in meat and liver.

[The effects of the tangfukang capsules on cytokines of early diabetic nephropathy].

OBJECTIVE: To find out the mechanism of Tangfukang on the diabetic nephropathy(DN). METHOD: A Model of streptorotocin diabetic rats was used. The expressions of the AGEs, ICAM-1, IV-C and FN were observed in renal cortex of diabetic rats with immunohistochemical method. The level of the IL-1 beta in blood serum was measured by radioimmunoassay method and the level of TNF was determined in blood serum by ELISA method. Aminoguanidine was selected as the control medicine which could inhibit the expression of monenzymatic glycosylation end products of protein. All the results were compared to analyze the effect of Tangfukang on monenzymatic glycosylation of protein and cytokine expression in early diabetic nephropathy rats. RESULTS: Both Tangfukang and aminoguanidine significantly decreased the expression of AGEs, but Tangfukang was greatly superior to aminoguanidine in suppressing the expression of cytokine like ICAM-1, IL-1 beta, TNF-alpha and the ingredients of extracellular matrix like IV-C, FN. CONCLUSION: It may be the partical mechanism of Tangfukang to suppress the monenzymatic glycosylation of protein and to reduce the expression of cytokine in diabetes. The effect of Tangfukang is superior to that of aminoguanidine.