To study the chemical constituents of Dipsacus asper, chromatographic methods such as D101 macroporous resin, silica gel, octadecylsilyl (ODS) column chromatographic techniques and preparative HPLC were used, and five compounds were isolated from 70% (v/v) ethanol extract of the plant. By using spectroscopic techniques including 1H NMR, 13C NMR, 1H-1H COSY, HSQC, HMBC and TOF-MS, the compounds were identified as 3beta-hydroxy-24-nor-urs-4 (23), 12-dien-28-oic acid (1), ursolic acid (2), oleanolic acid (3), 3-O-alpha-L-rhamnosyl(1 --> 3)-beta-D-glucopyranosyl (1 --> 3)-alpha-L-rhamnosyl (1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranosyl ester (4), 3-O-[beta-D-xylopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 4)] [alpha-L-rhamnosyl(1 --> 3)]-beta-D-glucopyranosyl (1 --> 3)-alpha-L-rhamnosyl(1 --> 2)-alpha-L-arabinopyranosyl hederagenin (5), separately. Among them, 1 is a new compound, and 2 is isolated from this plant for the first time.
Dipsacaceae ; chemistry ; Molecular Structure ; Oleanolic Acid ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo investigate the effect of p65 gene inhibited by siRNA on neuronic differentiation in the marrow mesenchymal stem cells (MSCs).

METHODSThe MSCs were transfected with Rn-p65-siRNA. Fasudil hydrochloride induced MSCs differentiating into neurons. The non-transfected group and negative control group (transfected with negative control siRNA marked by Cy3) were used as controls. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope at 24 h,48 h and 72 h after transfected with negative control siRNA. The viability of MSCs was detected by MTT at 24 h, 48 h and 72 h after transfected with Rn-p65-siRNA. The expressions of p65 mRNA and protein in MSCs were detected by RT-PCR and Western blot respectively. The expressions of p65 protein, NSE, MAP-2 and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method after transfection for 6 h.

RESULTSThe fluorescence of MSCs was mostly displayed after transfection of 72 hours and the efficiency of transfection was up to 83.3% ± 3.8%. Meanwhile, the p65 mRNA and p65 protein expressed by MSCs of transfected group were significantly decreased (P < 0.05); MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). The best efficiency of induction was observed in the transfected group. There were higher expressions of NSE and MAP-2 than the other group (P < 0.05).

CONCLUSIONThe p65 gene inhibited by siRNA can promote the marrow mesenchymal stem cells to differentiate into neurons.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; Animals ; Cell Differentiation ; Glial Fibrillary Acidic Protein ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; RNA, Messenger ; RNA, Small Interfering ; Rats ; Transcription Factor RelA ; antagonists & inhibitors ; metabolism ; Transfection

OBJECTIVETo investigate the significance of PKC and p44/42 mitogen-activated protein kinase (MAPK) signal transduction in ischemic preconditioning (IP).

METHODSThrough liver cell IP models, PKC inhibitor and MEK inhibitor were utilized to analyze the phosphorylation level of p44/42 MAPK and cell viability was also observed. Rat liver IP models were established which were treated with various drugs. Then the phosphorylation level of p44/42 MAPK in vivo and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) concentrations were detected. And cellular structures were observed under light microscopy.

RESULTSSimilar results were obtained in vivo and in vitro IP models. Compared with the ischemia reperfusion (IR) group in vivo, the phosphorylation level of p44/42 MAPK was obviously increased in IP treated rats (q = 27.217, P < 0.01), and the cellular structure injured slightly. The concentrations of serum ALT and AST in IP group were significantly lower than those in IR group (281.0 U/L +/-35.6 U/L vs 762.8 U/L +/-130.5 U/L and 407.7 U/L +/-73.7 U/L vs 820.9 U/L +/-111.3 U/L, P < 0.01). However, opposite changes were found in PKC and MEK inhibited groups, when compared to IP group. The phosphorylation level of p44/42 MAPK was obviously decreased, the liver tissues injured evidently, and the concentrations of serum ALT and AST (645.61 U/L +/-90.4 U/L, 678.6 U/L +/-136.5U/L and 466.2 U/L +/-82.8 U/L, 732.9 U/L +/-91.1 U/L, respectively) were significantly greater than those in IP group.

CONCLUSIONThese results suggest that p44/42 MAPK pathway plays a vital role in the protection of hepatocytes in ischemic preconditioning.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Hepatocytes ; cytology ; physiology ; Humans ; Ischemic Preconditioning ; Liver ; blood supply ; cytology ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVETo evaluate the clinical effects of carotid endarterectomy for carotid stenosis and occlusion.

METHODSFrom August 2005 to November 2008 moderate and severe carotid stenosis or occlusion were found in 16 patients by Doppler ultrasonography (DUS), MRA, CTA, DSA. The stenosis degree ranged from 60% to 99% in 14 patients and complete occlusion in 2 patients. Twelve patients underwent standard carotid endarterectomy (sCEA) in whom 2 patients were placed carotid shunt and 1 patient underwent carotid patch angioplasty. Four patients underwent eversion carotid endarterectomy (eCEA). All operations were performed by microscope.

RESULTSThere was no stroke, transient ischemic attack and mortality perioperatively and during follow-up from 1 month to 3 years. The ICA flow detected by follow-up duplex scan and MRA was unobstructed. The primary cerebral ischemic symptoms were obviously improved or disappeared after operation. The postoperative complications included one case of upper gastrointestinal hemorrhage and one case of hoarseness and bucking, which disappeared after medical treatment.

CONCLUSIONSCEA is an effective way for treating carotid stenosis. Different operative methods and techniques deal with different carotid lesions to achieve better effect. Microsurgical technique is useful for exposure of high ICA bifurcation and avoid effectively cranial nerve injury and other complications.

Adult ; Aged ; Carotid Stenosis ; surgery ; Endarterectomy, Carotid ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; Middle Aged ; Treatment Outcome

OBJECTIVESTo evaluate the clinical significance of flow cytometry (FCM) to detect the cytomegalovirus (CMV) PP65 antigen in patients with CMV infection.

METHODSSamples from 35 patients without CMV infection were used as negative control. The definite diagnosis of CMV infection was based on the national criteria for CMV infection. All 136 patients with CMV infection were examined with the FCM to detect CMV PP65 antigen, real-time fluorescence quantitative-polymerase chain reaction assay (RFQ-PCR) to detect CMV-DNA and ELISA to measure the serum level of IgM antibody against CMV. The results of these 3 assays in 2 groups (isolated organ involvement and disseminated diseases) were compared and the significance of PP65 antigenemia was evaluated. A short-term follow-up was undertaken in 18 patients.

RESULTSThe percentages of PP65 positivity in blood mononuclear cells (MNC) and polymorphic nuclear leukocyte (PMNL) from 35 negative control patients were 0.21% +/- 0.09% with a range of 0 - 0.41% and 0.24% +/- 0.10% with a range of 0.12% - 0.48%, respectively, which were not significantly different (t = 0.425, P > 0.05). The 95(th) percentiles (P(95)) of PP65 in MNC and PMNL were 0.39% and 0.45%, respectively, so a cutoff value of >/= 0.50% was set. Of the 136 patients with CMV infection, 118 samples from 118 patients were positive for PP65 antigenemia with a positive rate of 86.8%, which was not statistically different from that (90.4%, chi(2) = 0.91, P > 0.05) of CMV-DNA detected by RFQ-PCR assay but it was significantly higher than that (45.6%, chi(2) = 51.50, P < 0.005) of the detection by IgM measurement. PP65 detection was correlated with urine CMV DNA amplification (chi(2) = 63.78, P < 0.01) while the different detection rates between the two assays were not statistically significant (chi(m)(2) = 1.78,P > 0.05). PP65 detection was not correlated with serum IgM measurement while the detection rates between the two were significantly different (chi(m)(2) = 52.92,P < 0.01). No significant difference was found between the detection rates of CMV infection in MNC (45/53, 84.9%) and PMNL (43/53, 81.1%) (chi(m)(2) = 0.25, P > 0.05). Higher PP65 antigenemia level was correlated with systemic CMV infection, while lower level of PP65 was either in the patients with isolated organ involvement by CMV (chi(2) = 38.51, P < 0.005) or less severe in patient's situation. PP65 antigenemia of CMV infection returned to lower level or negative in recovery stage and increased when condition of patients deteriorated.

CONCLUSIONSPP65 antigenemia detection by FCM is effective in the diagnosis of the active CMV infection. Quantitative monitoring of PP65 antigenemia is useful in the evaluation of patients with CMV infection.

Antigens, Viral ; analysis ; Cytomegalovirus ; immunology ; Cytomegalovirus Infections ; diagnosis ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Immunoglobulin M ; analysis ; Polymerase Chain Reaction

OBJECTIVESTo explore the therapeutic effects of metformin on rat fatty livers induced by high fat feeding.

METHODSA fatty liver model was established by feeding rats with a high caloric laboratory chow for 12 weeks, then the rats were randomly divided into three groups, i.e. model control group, metformin group and dietary treatment group. A normal control group was organized at the same time. The rats of the metformin group were given metformin 156 mg/kg/d while the other groups were given distilled water of the same volume by stomach feeding. The model control group rats were fed with high caloric laboratory chow while other groups were fed a normal diet. After four weeks, all the animals were sacrificed. Liver index (liver/body weight ratio), serum activities of liver-associated enzymes, blood lipids, liver triglycerides, fasting blood glucose, fasting plasma insulin, HOMA insulin resistance index (HOMA-IR), and the liver histology of rats of all groups were assayed.

RESULTSThe body weight, liver index, serum activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), total cholesterol (TC), triglycerides (TG) and liver triglycerides in the model group increased significantly, while HDL-cholesterol concentration decreased significantly. Fasting blood glucose, fasting plasma insulin and HOMA-IR showed an increasing tendency, but there was no significant difference of those indexes among the three groups. The liver histology in the model group showed moderate to severe steatosis, mainly as macro vesicle steatosis, lobular inflammatory, cell infiltration and necrosis. Compared with the model group, the levels of body weight, liver index, serum ALT, ALP, TC, TG and liver triglycerides in the metformin group were significantly lower and were similar to those of the normal group, while their HDL-cholesterol concentration was significantly higher. The liver histology in the metformin group was nearly normal. In the dietary treatment group, hyperlipidemia persisted, although liver index and GGT were lower and the liver histology changes were somewhat milder.

CONCLUSIONIt is suggestive that metformin might be effective in treating rat fatty liver induced by high fat feeding.

Animals ; Dietary Fats ; administration & dosage ; Fatty Liver ; drug therapy ; etiology ; Hypoglycemic Agents ; therapeutic use ; Male ; Metformin ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of Rhizoma Coptidis and Radix Rehmanniae with the different ratio on the pharmacokinetics of berberine in rats.

METHOD24 rats were grouped to 4 groups randomly. Decoction, in which the proportion of Rhizoma Coptidis to Radix Rehmanniae is 1:0, 1:1, 1:4, 1:8 differently, was intragastrically given to the 4 groups. HPLC was used to determine concentration of berberine in serum. We adopted DAS 2.0 to analysis pharmacokinetic parameters of berberine.

RESULTThe concentration-time curves was all fitted to two-compartment model with a weight of 1/C2. Difference of 4groups in C(max), AUC(0-t), AUC(0-infinity), is significant (P <0.05).

CONCLUSIONRadix Rehmanniae of large dose can effectively enhance berberine's bioavailability in rats.

Animals ; Area Under Curve ; Berberine ; blood ; chemistry ; pharmacokinetics ; Biological Availability ; Coptis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacokinetics ; Male ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rehmannia ; chemistry ; Rhizome ; chemistry

OBJECTIVETo investigate the effect of notch signaling on differentiation of rat bone marrow mesenchymal stem cells (MSCs) into neurons induced by fasudil hydrochloride.

METHODSThe experiments were divided into non-transfected group, transfected group (transfected with Rn-Notch1-siRNA), positive control group (transfected with Rn-MAPK-1 Control siRNA) and negative control group (transfected with negative control siRNA). Fasudil hydrochloride induced MSCs differentiating into neurons. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope. The expression of notch1 mRNA, Hes1 mRNA and MAPK1 mRNA in MSCs was detected by RT-PCR. The expression of Notch1 protein, NSE, neurofilament M (NF-M) and glial fibrillary acidic protein(GFAP)was detected by immunocytochemical method. The viability of MSCs was detected by MTT.

RESULTS(1) The fluorescence of MSCs was mostly displayed after transfection for 72 h and the efficiency of transfection was up to 91.3% +/- 4.2%. Meanwhile, the notch1 mRNA and Hes1 mRNA expressed by MSCs of transfected group were significantly decreased (P < 0.05) and MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). (2) Fasudil hydrochloride could induce MSCs differentiate into neurons and the best efficiency of induction was observed in the transfected group. There was higher expression of NSE and neurofilament-M (NF-M) than the other groups (P < 0.05).

CONCLUSIONThere may be notch1 signaling and Rho/Rho GTPase signaling synergy on differentiation of rat bone marrow stromal cell into neurons induced by fasudil hydrochloride and they jointly promote the differentiation of MSCs into neurons.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; pharmacology ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Neurons ; cytology ; Rats ; Rats, Wistar ; Receptor, Notch1 ; metabolism ; Signal Transduction

Objective To understand the contamination status of common food-borne pathogens in foods sold in Changzhou, and to provide evidence for food safety risk assessment and prevention of food-borne diseases. Methods From 2010 to 2020 , 2 513 samples of 17 types of foods were collected in Changzhou area. The detection of pathogenic bacteria was carried out in accordance with the standard operation procedure specified in the ^“Workbook for Surveillance on Food Microorganisms and Pathogenic Factors in Jiangsu Province^”. Results A total of 260 positive samples of common food-borne pathogens were detected in all 2 513 samples with an overall detection rate of 10.30%. Single factor analysis showed that the detection rate of pathogenic bacteria in non-ready-to-eat samples was higher than that in ready-to-eat samples (χ²=148.875，P =0.000). The detection rate of pathogenic bacteria in bulk samples was higher than that in prepackaged ones (χ²=70.956，P=0.000). There is a difference in the detection rate of food-borne pathogens from different types of sampling sites (χ²=65.017，P=0.000). Logistic regression analysis showed that ready-to-eat food, packaging type, and sampling season were significantly correlated with the detection rate of food-borne pathogens. The detection rate of pathogenic bacteria in samples collected in the third or fourth quarters was higher than that in the first quarter. Conclusion The commercial foods sold in Changzhou have a relatively high level of contamination of food-borne pathogenic bacteria, and they should be fully heated and sterilized before consumption. The relevant departments should strengthen supervision and health education in summer and autumn.