HPLC fingerprint studies and determination of six components in Phyllanthus urinaria L. collected from different habitats / 国际中医中药杂志

Objective:To establish a method of measuring the contents of gallic acid, brevifolin, corilagin, geraniin, ellagic acid and rutin in Phyllanthus urinaria L. simultaneously with fingerprint study for analysis. Methods:Phyllanthus urinaria L. was extracted by ultrasound with 50% methanol. Chromatographic separation was performed on a Phenonmenex Luna C18 (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitrile (A) and 0.1% phosphoric acid aqueous solution (B) with gradient elution. The flow rate was 1.0 ml/min. The column temperature was 25 ℃, and the injection volume was 10 μl. The detection wavelength was 270 nm. HPLC fingerprints of Phyllanthus urinaria L. from different habitats was established. PCA and OPLS-DA were used to analyze the differences in chemical components of different habitats. Results:Gallic acid, brevifolin, corilagin, geraniin, ellagic acid and rutin showed good linearity at 0.042 8-0.641 6, 0.033 4-0.501 4, 0.142 2-2.133 1, 0.383 1-5.746 5, 0.063 1-0.946 2 and 0.019 2-0.287 8 μg, respectively. The average recovery rate of them was 103.65%, 96.39%, 101.85%, 95.04%, 98.79% and 98.33%, respectively. The HPLC fingerprints of different habitats contained 14 characteristic common peaks, and six compounds characteristic peaks were identified. PCA analysis showed that the chemical components of Phyllanthus urinaria L. from different habitats were different. Geraniin, ellagic acid and corilagin were screened by OPLS-DA. Conclusions:The method is efficient, accurate and sensitive, which can be used to measure the six components in Phyllanthus urinaria L.. The established HPLC fingerprint of different habitats combined with the measrurement method of six components can be used for the quality control and evaluation of Phyllanthus urinaria L..

Differential proteomic analysis of children infected with respiratory syncytial virus

Respiratory syncytial virus (RSV) infection is the main cause of lower respiratory tract infection in children. However, there is no effective treatment for RSV infection. Here, we aimed to identify potential biomarkers to aid in the treatment of RSV infection. Children in the acute and convalescence phases of RSV infection were recruited and proteomic analysis was performed to identify differentially expressed proteins (DEPs). Subsequently, promising candidate proteins were determined by functional enrichment and protein-protein interaction network analysis, and underwent further validation by western blot both in clinical and mouse model samples. Among the 79 DEPs identified in RSV patient samples, 4 proteins (BPGM, TPI1, PRDX2, and CFL1) were confirmed to be significantly upregulated during RSV infection. Functional analysis showed that BPGM and TPI1 were mainly involved in glycolysis, indicating an association between RSV infection and the glycolysis metabolic pathway. Our findings provide insights into the proteomic profile during RSV infection and indicated that BPGM, TPI1, PRDX2, and CFL1 may be potential therapeutic biomarkers or targets for the treatment of RSV infection.

Molecular Identification of Taenia hydatigena from Sheep in Khartoum, Sudan

The cestode Taenia hydatigena uses canids, primarily dogs, as definitive hosts, while the metacestode larval stage cysticercus infects a range of intermediate hosts, including domestic animals such as goats, sheep, and pigs. Cysticercosis due to T. hydatigena has large veterinary and economic drawbacks. Like other taeniids, e.g., Echinococcus, intraspecific variation is found among the members of the genus Taenia. In Africa, few studies are available on the epidemiology and distribution of T. hydatigena, and even fewer studies are available on its genetic variation. In this study, we molecularly identified 11 cysticerci from sheep in Sudan and demonstrated the genetic variation based on the NADH dehydrogenase subunit 1 (nad1) and cytochrome c oxidase subunit 1 (cox1) mitochondrial genes. The isolates were correctly identified as T. hydatigena with more than 99% similarity to those in the GenBank database. Low diversity indices and insignificant neutrality indices were observed, with 3 and 2 haplotypes for the nad1 and cox1 genes, respectively. The results suggest the presence of unique T. hydatigena haplotypes in Sudan, as haplotypes with 100% similarity were not found in the GenBank database. With few available studies on the genetic variation of T. hydatigena in Africa, this report represents the first insights into the genetic variation of T. hydatigena in Sudan and constitutes useful data.

Study on the current situation and allocation equity of China's nursing human resources / 中国卫生政策研究

Objective:To analyze the current situation of Human resources distribution of registered nurses in China, this study aims at providing evidence for the government departments to enact proper strategies from the aspects of quantity, structure and distribution equity, and to explore the problems existing in the nurturing of nursing talents. Methods:Descriptive analysis was used todescribe the quantity and quality of the registered nurse personnel during the period from 2010 to 2015, and the configuration fairness was evaluated by Gini Coefficient according to the population and size of geographical areausing data from China Health Statistics Yearbook and China Health and Family Planning Statistics Yearbook. Results :The results showed that from 2010 to 2015, the number of registered nurse personnel presented an increasing trend, and the personnel structure was further optimized and showed an overall trend of younger. From the perspective of distribution equity, the China's nursing personnel have a Gini coefficient which is less than 0. 2, and this highlights a good fairness according to the distribution of the population ; the Gini coefficient of the geographical area allocation was a-round 0.6, which shows great disparity. Conclusions :Remarkable results have been achieved in nursing human resource construction in China,but there are also many problems such as unreasonable nursing talent echelon and the uneven development across regions cannot be ignored too. It is recommended to expand the scaleof nurse training, strengthen the nursing education, improve the nursing efficiency and reasonably allocate the nursing human resources.

Phased Expression of Gene and Protein of NogoA and NgR in Rats with Spinal Cord Injury and Time Window of Electroacupuncture / 中国康复理论与实践

Objective To investigate the phased expression of gene and protein of NogoA and its receptor (NgR) that affects axon growth of spinal cord injury (SCI), and to explore the time window effect of electroacupuncture on SCI rats. Methods A total of 144 female Sprague-Dawley rats were randomly assigned to sham operation group (group A, n=48) and model group (n=96). In the model group, Allen's method was used to establish SCI rats model, and they were further subdivided into model control group (group B, n=48) and electroacupuncture group (group C, n=48). Group C received electroacupuncture on Dazhui (GV14), Yaoyangguan (GV3), bilateral Ciliao (BL32) and Zu-sanli (ST36) with loose-tight wave, for 20 minutes every day, one day, seven days and 14 days after modeling. The rats at every interventional therapy time were randomly subdivided into two subgroups, which accepted sev-en or 14 days of treatment. Groups A and B were killed and the injured spinal cord tissue was extracted one day, three days, seven days, 14 days and 28 days after modeling, group C at the corresponding time. The hind limb motor function was assessed with BBB score before all of rats were killed. Four samples at every time in each group were randomly selected to detect the expression of mRNA and protein of NogoA and NgR at different stage of SCI using reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. Results The BBB score began to increase 14 days after modeling, and significantly increased until 28 days after model-ing (P<0.05), compared with one day, three days and seven days after modeling in group B. The BBB score in-creased in group C than in group B at all the time points (P<0.05), except 14 days after electroacupuncture one day after modeling. The BBB score was higher as electroacupuncture intervening seven days and 14 days after modeling than that at one day after modeling in group C, and no significant difference was found between seven days and 14 days of treatment at either electroacupuncture time point (P>0.05). The expression of gene and pro-tein of NogoA and NgR in group B was in the increasing tendency after SCI, and was at the peak until 21 days af-ter modeling, and was higher in group B than in group A at each time point (P<0.01). The expression of gene and protein of NogoA decreased at all the time points in group C than in group B (P<0.05), except seven days of elec-troacupuncture intervening one day after modeling in the expression of NogoA mRNA (P>0.05). The expression of gene and protein of NogoA and NgR was lower as electroacupuncture intervening 14 days after modeling than one day after modeling in group C (P<0.05). There was no significant difference in the expression of gene and protein of NogoA and NgR between electroacupuncture intervening 14 days and seven days after modeling, and seven days and one day after modeling (P>0.05); as well as between seven days and 14 days of treatment at each time point (P>0.05). Conclusion Elerctroacupuncture could improve the hind limb motor function, which may associate with the inhibition of the expression of gene and protein of NogoA and NgR in injured spinal cord of rats after SCI. Elerctroacu-puncture is effective in the treatment of SCI at the early time, however, it is much better in the recovery stage.

Research on HPLC Fingerprint of Jinsangqi Kangdu Dropping Pills / 中国中医药信息杂志

Objective To establish fingerprint of Jinsangqi Kangdu Dropping Pills by HPLC; To control the quality of the preparation. Methods Waters XSELECT CSH-C18 chromatographic column (4.6 mm × 150 mm, 5 μm) was used and eluted with acetonitrile - 0.1% phosphoric acid solution gradient at the flow rate of 1.0 mL/min. The detection wavelength was 260 nm with column temperature of 30 ℃. Using calycosin-7-O-β-D-glucopyranoside, rutin, liquiritin, hyperoside, quercetin and ammonium glycyrrhizinate as the object references, ten batches of Jinsangqi Kangdu Dropping Pills were tested and analyzed by similarity comparison.Results Fringerprint spectrum of Jinsangqi Kangdu Dropping Pills had 24 common peaks in total, and characteristic spectrums of Hypericum Perforatum, Mori Cortex, Astragali Radix and Glycyrrhizae Radix et Rhizoma had been found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can evaluate the Jinsangqi Kangdu Dropping Pills quality totality,which can provide references for improving the quality control of the preparation.

Discussion on Commodity Specification and Grade Standard of Radix and Rhizoma of Chinese Materia Medica / 中国中医药信息杂志

Through literature review and field investigation, the commodity specification, grade standard and application situation of radix and rhizoma of Chinese materia medica were analyzed, and research methods and achievements of radix and rhizoma of Chinese materia medica were concluded, the existing problems of radix and rhizoma of Chinese materia medica were analyzed and concluded, and proposed identification methods fully referring Chinese methods of radix and rhizoma of Chinese materia medica. By combining modern planting and initial processing methods, classification indicators were specifically quantified, grade standard radix and rhizoma of Chinese materia medica conforming to current pharmacopoeia and meeting practical operation was designed, which were in accordance with market demand for high quality and high price standard system.

Optimization of the conditions for the synthetic preparation of iron sucrose complex by response surface methodology / 国际药学研究杂志

Objective To optimize the conditions for the synthetic process of iron sucrose complex (ISC), via the investigation of the effects of reaction temperature (X1), reaction time (X2), amount of alkali (X3), and amount of sucrose (X4) on the relative molecular mass of the ISC product. Methods According to the experimental results for the single factor, the conditions dealing with the X1, X2, X3, and X4 parameters for the preparation of ISC were optimized by the Box-Behnker design combined with the response surface methodology using the weight average relative molecular mass of ISC as an indicator, and analyzed with gel permeation chromatography. Results The reaction temperature and the amount of alkali had a significant effect on the weight average relative molecular mass of ISC. The influence of the four factors in the descending order was as follows:X3＞X1＞X2＞X4. In the designed experimental conditions, theresponsevaluedecreasedwiththeincreaseofbothreactiontemperaturesandalkaliamounts. Conclusion Theresponse surface methodology could provide the relationship between the response values and variables via the minimum number experiments to obtain the optimized conditions for the preparation of ISCs.

Relationship between oxidative stress and inflammation in peripheral and cerebral system of oxonate-induced hyperuricemic rats

To study what kind of role uric acid play on the relationship between oxidative Stress and inflammation in peripheral and cerebral system of oxonate-induced hyperuricemic rats. Twenty-six eight male Wistar rats were divided into two groups randomly. Potassium oxonate was used to establish hyperuricemic model for four weeks. In 2nd and 4th week, uric acid (UA) level, total superoxide dismutase (T-SOD), Gu,Zn-SOD activity and interleukin-1beta (IL-1ß) concentration in serum were determined respectively. In 4th week, one hour after last PO treatment, five rats of every group were given Evans Blue to test blood-brain barrier (BBB) permeability. Other brains were obtained to analysis T-SOD, Gu,Zn-SOD activity and IL-1ß concentration in cerebral system. Meanwhile, brain and kidney were stained with hematoxylin and eosin (H&E) to observe pathological change. In 2nd week, both of T-SOD and Gu,Zn-SOD activity in serum increased obviously (P<0.05) in hyperuricemia rats. However, IL-1ß content didn't change remarkably. In the 4th week, T-SOD activity in model group had become similar with control group, and at the same time IL-1ß content in serum increased significantly (P<0.05). Pathological section showed the structural and functional unit of the kidney had been damaged. On the contrary, both of T-SOD and Gu,Zn-SOD activity in brain increased obviously (P<0.05), but IL-1ß concentration was no significant difference between two groups. In addition, the results of Evans Blue and H&E suggested the integrity of BBB and structure of brain were not changed after PO treatment. The permeability of BBB and form of UA would be potential factors to decide what kind role UA play on keeping balance between anti-oxidative stress and induction of inflammatory response.

Diagnosis and treatment of patients with bile duct injury after laparoscopic cholecystectomy / 中华肝胆外科杂志

Objective To study the role of endoscopic retrograde cholangiopancreatography (ERCP) in diagnosing and treating iatrogenic bile duct injury after laparoscopic cholecystectomy (LC).Methods A retrospective study was conducted on 45 patients with iatrogenic bile duct injury after LC who were investigated and treated by ERCP from December 2002 to August 2015.Results Using the StrasbergBismuth classification,there were 14 patients with type A and 4 with type C who were managed successfully using endoscopic nasobiliary drainage (ENBD) and interventional ultrasound abdominal localized puncture and drainage ; 7 patients with type D were managed successfully using endoscopic sphincterotomy (EST) and endoscopic retrograde biliary drainage (ERBD).For the 5 patients with type E Ⅰ and 3 patients with type E Ⅱ who were treated by EST and ERBD,one patient who had common bile duct transection required cholangioenteric Roux-en-Y anastomosis.For the 6 patients with type E Ⅲ and 6 patients with type EⅣ who were treated by EST and ERBD,a patient required a cholangioenteric Roux-en-Y anastomosis to achieve good results.Conclusions When iatrogenic bile duct injury is suspected after LC,correct assessment with ERCP should be taken immediately.ERCP when combined with ENBD and (or) ERBD could reduce bile duct pressure and dilate stenotic bile ducts to avoid further operation.

HPLC fingerprint of analgesic ingredients in cangbaiqutong capsules / 国际药学研究杂志

Objective To establish fingerprint of analgesic ingredients in Cangbaiqutong capsules by HPLC, and control different production batches of Cangbaiqutong capsules’ quality. Methods Waters XSELECT CSH-C18 (4.6 mmxl50 mm, 5.0μm) chromatographic column was used and eluted with acetonitrile -0.1% phosphoric acid solution gradiently in the flow rate of 1.0 ml/ min. The detection wavelength was 284 nm with column temperature of 30°C. rrith phellodendrine chloride, gentiopicroside, paeoniflorin, tetrandrine and berberine hydrochloride as the object references, ten batches of Cangbaiqutong capsules were tested and analyzed by similarity comparison. Results fingerprint spectrum in Cangbaiqutong capsules had 20 common peaks in total, and characteristic spectra of Cortex Phellodendri, Gentiana, Radix Paeoniae Rubra and Radix Stephaniae Tetrandrae were found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can be used to evaluate the Cangbaiqutong capsules% quality in the round, which could improve the quality control standard in productive process of Cangbaiqutong capsules.

Study on the Quality Standard for Cangbai Qutong Capsule / 中国药房

OBJECTIVE:To establish the quality standard for Cangbai qutong capsule. METHODS:TLC was conducted to iden-tify the Rhizoma atractylodis,Asarum sieboldii,Paeoniae Radix Rubra and Cortex phellodendri;RP-HPLC was conducted to deter-mine the content of berberine hydrochloride. The column was Waters Symmetry-C18 with mobile phase of acetonitrile-0.1% phos-phoric acid (gradient elution) at flow rate of 1.0 ml/min,the detection wavelength was 265 nm,column temperature was 30 ℃, and the injection volume was 10 μl. RESULTS:TLC showed R. atractylodis,A. sieboldii,Paeoniae Radix Rubra and C. phelloden-dri had clear spots and good separation. The linear range of berberine hydrochloride was 21.12-84.48 μg/ml(r=0.999 6);RSDs of pre-cision,stability and reproducibility tests were lower than 3%;recovery was 96.439%-100.950%(RSD=1.34%,n=9). CONCLU-SIONS:The method is simple and reproducible,and can be used for the quality standards of Cangbai qutong capsule.

HPLC fingerprint of analgesic ingredients in Cangbaiqutong capsules / 国际药学研究杂志

Objective To establish fingerprint of analgesic ingredients in Cangbaiqutong capsules by HPLC, and control different production batches of Cangbaiqutong capsules′ quality. Methods Waters XSELECT CSH-C18 (4.6 mm×150 mm, 5.0μm) chromatographic column was used and eluted with acetonitrile -0.1% phosphoric acid solution gradiently in the flow rate of 1.0 ml/min. The detection wavelength was 284 nm with column temperature of 30℃. rrith phellodendrine chloride, gentiopicroside, paeoniflorin, tetrandrine and berberine hydrochloride as the object references, ten batches of Cangbaiqutong capsules were tested and analyzed by similarity comparison. Results fingerprint spectrum in Cangbaiqutong capsules had 20 common peaks in total, and characteristic spectra of Cortex Phellodendri, Gentiana, Radix Paeoniae Rubra and Radix Stephaniae Tetrandrae were found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can be used to evaluate the Cangbaiqutong capsules′quality in the round, which could improve the quality control standard in productive process of Cangbaiqutong capsules.

Research on Extraction Process of Zhechong Chuangyu Capsule / 中国中医药信息杂志

Objective To establish the extraction process of Zhechong Chuangyu Capsule. Methods The difference of analgesic effect of water extraction and alcohol extraction in mice was observed by body-torsion test to determine the extract solvent. With the rate of aqueous extraction, n-butyl alcohol extraction and asperosaponin Ⅵ as evaluating indicator, the influencing factors including solvent volume, time and times of extraction were investegated to evaluate extracting procedure by orthogonal experiment. Results There was no obvious difference in analgesic effect between water extraction and alcohol extraction. Given the requirement of produce, aqueous extraction was a better choice. The optimum extracting condition was extracted 3 times with 20 folds volume of solvent, and extraction time was 150 minutes. Conclusion The extraction process is feasible to be applied into production.

Preliminary analysis of taenia multiceps metacestode antigens by two-dimensional electrophoresis

Taenia multiceps is a cestode parasite with its larval stage [metacestode], Coenurus cerebralis, mainly encysts in the central nervous system of sheep and other livestock causing cerebralis coenurosis. Since treatment of coenurosis with chemotherapy showed little effect and surgical removal of cysts is not advisable in field conditions, vaccination is useful to control coenurosis. Previous study indicated that immunization with T. multiceps metacestode antigens could induce protection in sheep against coenurosis, so the aim of this study was to identify T. multiceps metacestode antigens in order to find potential vaccine development candidates for further study. The protein extracts from the larval T. multiceps were analyzed by twodimensional electrophoresis [2-DE] and characterized by mass spectrometry. A total of 150 protein spots were detected with isoelectric point [pI] value from 4.97 to 9.65 and molecular weight from 14 to 98 kDa. Twenty-two protein identities were determined by mass spectrometry and 15 unique proteins were obtained. Functional annotation revealed that some of these proteins are involved in catalytic activity, binding, metabolic, cellular process and stress response. Among these molecules are antioxidant proteins [peroxiredoxin and glutathione-S-transferase], glycolytic enzymes [malate dehydrogenase and enolase], proteins with chaperone activity [heat shock protein 70 and small heat shock protein], and structural proteins [actin, actin modulator protein and paramyosin]. The identification of T. multiceps metacestode protein will provide valuable information to elucidate their specific roles in the parasitism and screen new targets for vaccine development

Molecular Characterization of Taenia multiceps Isolates from Gansu Province, China by Sequencing of Mitochondrial Cytochrome C Oxidase Subunit 1

A total of 16 Taenia multiceps isolates collected from naturally infected sheep or goats in Gansu Province, China were characterized by sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The complete cox1 gene was amplified for individual T. multiceps isolates by PCR, ligated to pMD18T vector, and sequenced. Sequence analysis indicated that out of 16 T. multiceps isolates 10 unique cox1 gene sequences of 1,623 bp were obtained with sequence variation of 0.12-0.68%. The results showed that the cox1 gene sequences were highly conserved among the examined T. multiceps isolates. However, they were quite different from those of the other Taenia species. Phylogenetic analysis based on complete cox1 gene sequences revealed that T. multiceps isolates were composed of 3 genotypes and distinguished from the other Taenia species.

Explore the protective effects of the bone marrow mesenchymal stem cells on severe acute pancreatitis in rats / 中华急诊医学杂志

Objective To investigate effects of bone marrow mesenchymal stem cells (MSC) on severe acute pancreatitis (SAP) in rats.Methods A total of 60 rats were randomly divided into normal group (n =10),the model of SAP group (n =10),the transplantation of the MSC group (n =20),the combination of the MSC and the granulocyte colony-stimulating factor (G-CSF) group (n =20).The conversion rate of MSC was detected by using immunofluorescence methods,and the level of amylase in serum was assayed by using biochemical methods.Simultaneously,the level of interleukin-6 (IL-6) in serum was determined by using enzyme linked immunosorbent assay (ELISA).Results The conversion rates of the MSC increased,and consequently,the levels of amylases and interleukin-6 in serum were reduced (P ＜ 0.05).When a small amount of the G-CSF was added to MSC,the therapeutic effects of the two different kinds of cells were synergistically strengthened.In the contrary,when a large number of the G-CSF was added to MSC,the antagonism resulted between these two different kinds of cells gives rise to harmful effects on SAP.Conclusions The bone marrow mesenchymal stem cells have therapeutic effects on SAP.When the number of the bone marrow mesenchymal stem cells increases,the protective effects are enhanced.

Research on bone marrow mesenchymal stem cells in the treatment of severe acute pancreatitis / 中华肝胆外科杂志

Objective To investigate the mechanism of acute inflammatory response and tissue repair when rats accepted transplanted bone marrow mesenchymal stem cells (MSC) in severe acute pancreatitis (SAP).Methods A total of 40 rats were randomly divided into 4 groups which included the normal group (n=10),the model severe acute pancreatitis group (n=10),the transplanted bone marrow mesenchymal stem cells group (n =10),and the combination of bone marrow mesenchymal stem cells and granulocyte colony-stimulating factor (G-CSF) group (n=10).To cure the acute severe pancreatic injury caused by SAP,rats were injected with EDU-labeled MSCs and granulocyte colonystimulating factor (Gr-CSF).The conversion rate of MSCs to pancreatic cells or MSCs to endothelial cells was detected to assess the role of MSCs in tissue regeneration and repair.The expression of amylase,interleukin-6 (IL-6),and interleukin-10 (IL-10) in serum was detected to assess the role of MSCs in an acute inflammatory response.Results The concentrations of amylase and IL-6 were reduced and the concentration of IL-10 was increased in MSC and MSC+G-CSF groups after the onset of SAP.Flow cytometry showed a small amount of MSCs converting to endothelial or pancreatic cells,but the conversion rate was relatively low.Conclusions MSCs can play an important role in the antipre-release of inflammatory mediators,reducing acute immune response to control the acute inflammatory response in SAP.Moreover,MSCs can repair a damaged pancreas by converting into both pancreatic and endothelial cells.

Garlicin ameliorated pressure overload induced myocardial fibrosis in rats through partial inhibiting TGF-beta1 mediated Smads signal / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe whether garlicin could ameliorate pressure overload induced myocardial fibrosis in rats through partial inhibiting transforming growth factor beta1 (TGF-beta1) mediated Smads signal.</p><p><b>METHODS</b>Forty male SD rats were randomly divided into 4 groups, i. e., the sham-operation group, the model group, the garlicin group, and the Tetramethylpyrazine (TMP) group, 10 in each group. The pressure overload induced myocardial fibrosis rat model was prepared using coarctation of aorta. Three days after modeling 5.0 mg/kg garlicin injection was administered to rats in the garlicin group, 20 mg/kg TMP injection to rats in the TMP group by peritoneal injection, while normal saline was peritoneally injected to rats in the sham-operation group and the model group. Four weeks after medication, the changes of myocardial collagen were observed by picrosirius red staining. The myocardial collagen volume fraction (CVF) and perivascular collagen areas (PVCA) were calculated. The serum transforming growth factor beta1 (TGF-beta1) expression was detected using ELISA. The TGF-beta1 protein expression in the myocardial tissue was observed using immunohistochemical assay. The changes of myocardial Smad2 and Smad7 mRNA expressions were detected using Real-time RT-PCR. The effects of garlicin on TGF-beta1 mediated Smad Signaling through luciferase assay were further verified using Mv1 Lu-(CAGA) 12-Luc cell line response to TGF-beta1.</p><p><b>RESULTS</b>Compared with the sham-operation group, the myocardial levels of CVF and PVCA, the serum TGF-beta1 level, and the TGF-beta1 protein expression in the myocardial tissue obviously increased in the model group (P < 0.05, P < 0.01). Compared with the model group, the PVCA level, the serum TGF-beta1 level, and the TGF-beta1 protein expression in the myocardial tissue of the garlicin group and the TMP group obviously decreased (P < 0.05, P 0O 01). The Smad2 mRNA expression was up-regulated while Smad7 mRNA expression down-regulated in the model group. The Smad2 mRNA expression was obviously down-regulated in the garlicin group and the TMP group (P < 0.05). The Smad7 mRNA expression was obviously up-regulated in the TMP group (P > 0.05). One to 2 microg/mL garlicin could obviously inhibit the luciferase activities of corresponding TGF-beta1, under the stimulation of 2 ng/mL TGF-beta1 (P < 0.05).</p><p><b>CONCLUSION</b>Garlicin ameliorated pressure overload induced myocardial fibrosis in rats through partial inhibiting TGF-beta-Smads signal pathway.</p>

Genetic variation of the 8-kDa glycoprotein family from Echinococcus granulosus, Taenia multiceps and Taenia hydatigena / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Echinococcosis, coenurosis and cysticercosis are debilitating diseases which prevail in China. Immunological diagnosis of metacestodosis is important in disease control. The 8-kDa glycoproteins from taeniid cestodes have successfully been used for diagnosis of human cysticercosis in immunological assays. The aim of the present study was to investigate genetic variations and phylogenetic relationships of the 8-kDa proteins for evaluating the possibility of utilizing these proteins as diagnostic antigens for other metacestode infections.</p><p><b>METHODS</b>The genes and complementary DNAs (cDNAs) encoding the 8-kDa proteins from Echinococcus (E.) granulosus, Taenia (T.) multiceps and T. hydatigena were amplified using PCR method. Their amplicons were cloned into the vector pMD18 and the positive clones were sequenced. Sequence data were analyzed with the SeqMan program, and sequence homology searches were performed using the BLAST program. Alignments were conducted using the ClustalX program, and the phylogenetic analyses were performed with the Protein Sequences Program and the Puzzle Program using the Neighbor-joining method.</p><p><b>RESULTS</b>Fifteen, 18 and 22 different genomic DNA sequences were identified as members of the 8-kDa protein gene family from E. granulosus, T. multiceps and T. hydatigena, respectively. Eight, four and six different cDNA clones respectively from E. granulosus, T. multiceps and T. hydatigena were characterized. Analysis of these sequences revealed 54 unique 8-kDa protein sequences. Phylogenetic trees demonstrated that the taeniid 8-kDa proteins are clustered into eight clades at least: Ts18, Ts14, TsRS1, TsRS2, T8kDa-1, T8kDa-2, T8kDa-3 and T8kDa-4.</p><p><b>CONCLUSION</b>We found that the gene family encoding for the taeniid 8-kDa antigens is comprised of many members with high diversity, which will provide molecular evidence for cross-reaction or specific reaction among metacestode infections and may contribute to the development of promising immunological methods for diagnosis of metacestodosis.</p>