OBJECTIVE: To predict the anti-diabetes effects of Corydalis yanhusuo alkaloids with pharmacological network technology and verify the results in animal models. METHODS: Targets of Corydalis yanhusuo alkaloids were collected from Pubmed, CNKI, Wangfang and VIP databases, and targets for treatment of diabetes and diabetic complications were searched from OMIM database. The "components-targets-diabetes" network was constructed and analyzed with cytoscape 2.8.2. Diabetes model was established by STZ injection. RESULTS: The targets of Corydalis yanhusuo alkaloids covered many kinds of protein, including ion channel, G coupled protein, and signal proteins such as potassium channel, MAPK/ERK, VEGF and NOS3. In the network, Corydalis yanhusuo alkaloids associated with diabetes and diabetic complications, especially, non-insulin dependent diabetes, obesity, and diabetic complications via modulation of NOS3, VEGF, INSR, KCNJ11 and IRS1, indicating potential effects of Corydalis yanhusuo for diabetes and diabetic complications. Corydalis yanhusuo alkaloids decreased blood glucose in normal and diabetic ICR mice and improved sugar tolerance in insulin-resistant mice, which was in conformity with the prediction. CONCLUSION: Corydalis yanhusuo alkaloids show potential effects on diabetes and diabetic complications.

OBJECTIVETo understand the process of Cistanche tubulosa.

METHODThe process of seed germination and parasitism was observed using stereomicroscope.

RESULTSeedling of C. tubulosa sprouted after forty day without host root's contact in fields, a tube-like-organ formed and grew auger-type from host root, the tuber apex where touches host root swelled and formed haustorium. Haustorium intruded host root epidermis and vascular bundles, and released brown substances. Then, embryo bud with six or more young leaves formed, finally the swelled tuber-like-organ broken and seed coat shed. Due to the parasitism of C. tubulosa, the host root near stem site swelled, but the other part, shrunk and disappered gradually.

CONCLUSIONSeed of C. tubulosa could germinate indepently in fields. Tuber-like-organ formatin, haustorium formation and bud formation are key steps of C. tubulosa seedling development.

Cistanche ; growth & development ; Germination ; Plants, Medicinal ; growth & development ; Seeds ; growth & development ; Symbiosis ; Tamaricaceae ; growth & development

OBJECTIVETo study the curative effects of pirfenidone (PF) on pulmonary fibrosis induced by paraquat (PQ) in mice and to provide the theoretical basis for clinical treatment.

METHODSNinety adult healthy male ICR mice were randomly divided into six groups: control group, PQ group, 2 mg/kg Dexamethasone group, 25 mg/kg PF group, 50 mg/kg PF group and 100 mg/kg PF group, there were 15 mice in each group. The corresponding volume of normal saline was given to the each mouse in control group according to the weight, after 2 h 0.1% CMC was given to the each mouse of control group one time by intragastric administration, then the CMC was administrated at regular time until sacrifice. All mice for other 5 groups were exposed to 100 mg/kg PQ by intragastric administration. At 2 h after exposure to PQ, 0.02 ml/10 g dexamethasone and 25, 50, 100 mg/kg PF were given to mice for dexamethasone group and for 3 PF groups by intragastric administration each day for 49 days, respectively. The lung coefficient was calculated and pathological changes of lung tissue were observed by HE staining for each mouse. The hydroxyproline (HYP) level in lung tissue was measured for each mouse. The mRNA level of and the protein level of TGF-β(1) in lung tissue for each mouse were determined, and the protein level of TGF-β(1) in the bronchus-alveolus lavage fluid (BALF) of each mouse was detected.

RESULTSThe survival rates on the 3rd day in PQ group, 3 PF groups and dexamethasone group were 53.33%, 46.67%, 73.33%, 86.67% and 80%, respectively. The survival rates on the 3rd day in dexamethasone group, 50 mg/kg and 100 mg/kg PF groups were significantly higher than those of PQ group and 25 mg/kg PF group (P < 0.05). The lung coefficients of 3 PF groups were significantly lower than that of the PQ group (P < 0.05). The lung tissue HYP levels of dexamethasone group and 3 PF groups were 50.95 ± 11.65, 44.52 ± 9.48, 43.27 ± 6.01 and 40.82 ± 5.90 mg/g respectively, which were significantly lower than that (74.27 ± 3.68) of PQ group (P < 0.01). The TGF-β(1) protein levels of BALF in dexamethasone group, 50 and 100 mg/kg PF groups were 22.03 ± 7.27, 27.75 ± 5.84 and 21.31 ± 6.82 ng/ml respectively, which were significantly lower than that (52.52 ± 15.51) ng/ml of PQ group (P < 0.01) The expression level of TGF-β(1) mRNA in 100 mg/kg PF group decreased significantly, as compared with PQ group (P < 0.01).

CONCLUSIONPF could reduce the collagen deposition and pulmonary fibrosis induced by PQ in mice lungs.

Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Paraquat ; poisoning ; Pulmonary Fibrosis ; chemically induced ; drug therapy ; pathology ; Pyridones ; therapeutic use ; Transforming Growth Factor beta ; metabolism

Objective To analyze the region cluster and risk factors of hypertension in the Chinese adult population and to explore the application of multilevel regression model in the risk factors of hypertension. Methods Multi-stage random sampling technique was used to choose 15 540 individuals aged 35-74 years from 10 regions in China. Two-level logistic regression models were fitted under MLwiN 2.02 software. Results The region cluster of hypertension existed and variance portion coefficient was 3.1%. After adjusting for the age and gender, overall obese people (BMI≥28 kg/m2) were 4.50(95%CI: 4.00-5.06) times, overweight people (BMI=24-27.9 kg/m2) were 2.26 (95%CI: 2.07-2.46) times more likely to be hypertensive as compared with those of normal BMI (18.5-23.9 kg/m2), and those centrally obesive people (Waist circumference≥85 cm in male or 80 cm in female) were 2.62 (95%CI: 2.42-2.83) times more likely to be hypertensive as compared with those of normal WC. The age-and gender-adjusted odds ratios (Ors) of triglyceride (TG), serum total cholesterol (TC), glucose, low-density lipoprotein cholesterol (LDL-C) , high-density lipoprotein cholesterol (HDL-C) and drinking alcohol were 2.10 (95% CI: 1.89-2.33) , 2.08 (95% CI: 1.84-2.35) , 1.85 (95% CI: 1.60-2.14) , 1.58 (95% CI: 1.38-1.81), 1.49(95%CI: 1.32-1.69) and 1.15(95%CI: 1.05-1.27), respectively. Conclusion The prevalence of hypertension was not only affected by individual risk factors, such as obesity, drinking alcohol, abnormal glucose and serum lipids profile, but also affected by the geographic environment where people resided in. Population-and risk factors targeted strategies, proved a promising way to reduce individual risk of hypertension in the primary prevention of hypertension.

OBJECTIVETo discuss the clinical effectiveness in treating thoracolumbar fractures adopting the rehabilitation exercise utilizing knee pads on the orthopedic traction bed.

METHODSFrom June 1996 to June 2006, we studied the clinical effectiveness of thoracolumbar fractures utilizing knee pads on the orthopedic traction bed for rehabilitation exercise. The cases surveyed total 209, 163 of which had full data. There were 98 males and 65 females with the age from 17 to 74 years (mean, 14.5 years). Consulting time after injury from 30 min to 7 days. Fracture site in T11 had 8 cases, in T12 24 cases, in L1 73 cases, in L2 33 cases, in L3 8 cases, in L4 3 cases, in T12 and L1 14 cases. Compression degree of vertebral anterior border: full compression had 1 case,more than 4/5 had 23, more than 2/3 had 67, more than 1/2 had 40, in 1/3 had 46.

RESULTSAmong them, 8 cases with legs paresis no recovery in nerval function or stopping recovery changed methods, and underwent surgical treatment. Others 155 cases were followed up from 2 to 12 years with an average of 3 years and 4 months. The average height of vertebral anterior borders of the 169 injured compressed had increased from 1.55 cm before treatment to 2.70 cm after treatment with an average of 1.15 cm. The height of the injured vertebral anterior borders had recovered from 50.5% (1.55/3.07) before treatment to 89.4% (2.70/3.02) after treatment. Kyphosis angle of the injured vertebral bodies had recovered from 13.25 degrees to -1.6 degrees in average. Twenty-three cases associated with dislocation basic reduction.

CONCLUSIONRehabilitation exercise using knee pads on the orthopedic traction bed can obtain satisfactory clinical effect in treating thoracolumbar fractures, the method is easy. At 3, 7, 10 days after treatment, the height of bed should be adjusted according X-ray.

Adolescent ; Adult ; Aged ; Exercise Therapy ; instrumentation ; methods ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; injuries ; physiopathology ; surgery ; Male ; Middle Aged ; Orthopedic Equipment ; Recovery of Function ; Spinal Fractures ; physiopathology ; rehabilitation ; surgery ; Thoracic Vertebrae ; injuries ; physiopathology ; surgery ; Treatment Outcome ; Young Adult

BACKGROUNDH3N2 subtype influenza A viruses have been identified in humans worldwide, raising concerns about their pandemic potential and prompting the development of candidate vaccines to protect humans against this subtype of influenza A virus. The aim of this study was to establish a system for rescuing of a cold-adapted high-yielding H3N2 subtype human influenza virus by reverse genetics.

METHODSIn order to generate better and safer vaccine candidate viruses, a cold-adapted high yielding reassortant H3N2 influenza A virus was genetically constructed by reverse genetics and was designated as rgAA-H3N2. The rgAA-H3N2 virus contained HA and NA genes from an epidemic strain A/Wisconsin/67/2005 (H3N2) in a background of internal genes derived from the master donor viruses (MDV), cold-adapted (ca), temperature sensitive (ts), live attenuated influenza virus strain A/Ann Arbor/6/60 (MDV-A).

RESULTSIn this presentation, the virus HA titer of rgAA-H3N2 in the allantoic fluid from infected embryonated eggs was as high as 1:1024. A fluorescent focus assay (FFU) was performed 24-36 hours post-infection using a specific antibody and bright staining was used for determining the virus titer. The allantoic fluid containing the recovered influenza virus was analyzed in a hemagglutination inhibition (HI) test and the specific inhibition was found.

CONCLUSIONThe results mentioned above demonstrated that cold-adapted, attenuated reassortant H3N2 subtype influenza A virus was successfully generated, which laid a good foundation for the further related research.

Animals ; COS Cells ; Cercopithecus aethiops ; Dogs ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; Plasmids ; Reassortant Viruses ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, Attenuated ; immunology ; Viral Proteins ; genetics

Accumulating evidence has revealed that brain iron concentrations increase with aging, and the choroid plexus (CP) may be at the basis of iron-mediated toxicity and the increase in inflammation and oxidative stress that occurs with aging. The mechanism involves not only hepcidin, the key hormone in iron metabolism, but also iron-related proteins and signaling-transduction molecules, such as IL-6 and signal transducer and activator of transcription 3 (Stat3). The aim of the present study was to investigate the correlation between the IL-6/Stat3 signaling pathway and hepcidin at the CP in normal aging. Quantitative real time PCR and Western blot were used to determine the alterations in specific mRNA and corresponding protein changes at the CP at ages of 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33 and 36 months in Brown-Norway/Fischer (B-N/F) rats. The results demonstrated that hepcidin mRNA level at the CP kept stable in young rats (from 3 to 18 months), and increased with aging (from 21 to 36 months). The alterations of IL-6/p-Stat3 mRNA and protein expressions in normal aging were in accordance with that of hepcidin mRNA. Our data suggest that IL-6 may regulate hepcidin expression at the CP, upon interaction with the cognate cellular receptor, and through the Stat3 signaling transduction pathway.
Aging ; physiology ; Animals ; Choroid Plexus ; metabolism ; Hepcidins ; physiology ; Interleukin-6 ; physiology ; Iron ; metabolism ; RNA, Messenger ; Rats ; Rats, Inbred F344 ; STAT3 Transcription Factor ; physiology ; Signal Transduction

Accumulation of amyloid-beta peptides (Aβ) results in amyloid burden in normal aging brain. Clearance of this peptide from the brain occurs via active transport at the interfaces separating the central nervous system (CNS) from the peripheral circulation. The present study was to investigate the change of Aβ transporters expression at the choroid plexus (CP) in normal aging. Morphological modifications of CP were observed by transmission electron microscope. Real-time RT-PCR was used to measure mRNA expressions of Aβ(42) and its transporters, which include low density lipoprotein receptor-related protein-1 and 2 (LRP-1 and -2), P-glycoprotein (P-gp) and the receptor for advanced glycation end-products (RAGE), at the CP epithelium in rats at ages of 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33 and 36 months. At the same time, the mRNA expressions of oxidative stress-related proteins were also measured. The results showed that a striking deterioration of the CP epithelial cells and increased Aβ(42) mRNA expression were observed in aged rats, and there was a decrease in the transcription of the Aβ efflux transporters, LRP-1 and P-gp, no change in RAGE mRNA expression and an increase in LRP-2, the CP epithelium Aβ influx transporter. Heme oxygenase-1 (HO-1) and caspase-3 expressions at the CP epithelium increased with age at the mRNA level. These results suggest the efficacy of the CP in clearing of Aβ deceases in normal aging, which results in the increase of brain Aβ accumulation. And excess Aβ interferes with oxidative phosphorylation, leads to oxidative stress and morphological structural changes. This in turn induces further pathological cascades of toxicity, inflammation and neurodegeneration process.
ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Aging ; Amyloid beta-Peptides ; metabolism ; Animals ; Caspase 3 ; metabolism ; Choroid Plexus ; physiology ; Heme Oxygenase (Decyclizing) ; metabolism ; LDL-Receptor Related Proteins ; metabolism ; Oxidative Stress ; Peptide Fragments ; metabolism ; Rats ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism

OBJECTIVETo evaluate the efficacy and safety of a China made adefovir dipivoxil (ADV) treatment for hepatitis B e antigen-positive patients with chronic hepatitis B.

METHODSTwo hundred and thirty patients with chronic hepatitis B who were positive for hepatitis B e antigen (HBeAg) were randomly put into groups A or B, and 58 patients with lamivudine-resistant chronic hepatitis B were randomly put into groups C or D. During the first 12 weeks of the trial, 112 patients in group A and 115 patients in group B received 10 mg of ADV and a placebo once a day; 28 patients in group C received 100 mg of lamivudine (LMV) and 10 mg of ADV; 29 patients in group D received 100 mg of LMV and a placebo once a day. In the second trial period, all patients received ADV for 36 weeks. The primary checking criterion was the serum HBV DNA change during the treatment. The secondary ones were alanine aminotransferase (ALT) normalization, HBeAg loss, and HBeAg seroconversion.

RESULTSAt week 12, the median serum hepatitis B virus (HBV) DNA level of group A (ADV-ADV) was reduced 2.8 log10 copies/ml, significantly greater than that of group B (placebo-ADV) of 0.3 log10 copies/ml reduction (P = 0.000). At week 48, the median serum HBV DNA level of group A and group B were reduced 3.6 and 3.4 log10 copies/ml respectively. At week 12, the median serum HBV DNA level of group C (LMV+ADV) was reduced 3.0 log10 copies/ml, significantly greater than that of the group D (LMV+placebo) of 0.16 log10 copies/ml reduction (P = 0.000). At week 48, the median serum HBV DNA level of group C and group D were reduced 3.6 and 3.8 log10 copies/ml respectively. Only 5.56% (16/288) patients had adverse events that were mild to moderate. There was no significant difference in the change of serum creatinine compared with their baseline levels.

CONCLUSIONIn our HBeAg positive lamivudine-resistant chronic hepatitis B patients, 48 weeks of ADV treatment was safe and resulted in significant virological and biochemical improvements.

Adenine ; analogs & derivatives ; therapeutic use ; Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Double-Blind Method ; Drug Resistance, Viral ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; immunology ; virology ; Humans ; Middle Aged ; Mutation ; Organophosphonates ; therapeutic use ; Young Adult

Cardiac extracellular matrix (ECM), generated from the process of decellularization, has been widely considered as an ideal source of biological scaffolds. However, current ECM preparations are generally difficult to be applied to generate cardiac tissue. Our research was aimed to improve decellularization protocols to prepare cardiac ECM slices. Adult murine ventricular tissues were embedded in low melting agarose and cut into 300 μm slices, and then were divided randomly into three groups: normal cardiac tissue, SDS treated group (0.1% SDS) and SDS+Triton X-100 treated group (0.1% SDS+0.5% Triton X-100). Total RNA content and protein content quantification, HE staining and immunostaining were used to evaluate the removal of cell components and preservation of vital ECM components. Furthermore, murine embryonic stem cell-derived cardiomyocytes (mES-CMs) and mouse embryonic fibroblasts (MEFs) were co-cultured with ECM slices to evaluate biocompatibility. The relative residual RNA and protein contents of ECM slices significantly decreased after decellularization. HE staining showed that SDS+Triton X-100 treatment better destroyed cellular structure and removed nuclei of ECM slices, compared with SDS treatment. Immunostaining showed that collagen IV and laminin were better preserved and presented better similarity to original cardiac tissue in ECM slices acquired by SDS+Triton X-100 treatment. However, collagen IV and laminin were significantly decreased and arranged disorderly in SDS treated group. We observed effective survival (≥ 12 days) of MEFs and mES-CMs on ECM slices acquired by SDS+Triton X-100 treatment, and signs of integration, whereas those signs were not found in SDS treated group. We concluded that, compared with traditional SDS method, new combined protocol (SDS+Triton X-100) generated ECM slices with better component and structural preservation, as well as better biocompatibility.
Animals ; Extracellular Matrix ; chemistry ; Heart Ventricles ; cytology ; Mice ; Octoxynol ; Sodium Dodecyl Sulfate ; Tissue Engineering ; methods ; Tissue Scaffolds