Objective:To investigate the purification of antibacterial effective fraction of Syringae folium by macroporous resins. Methods:Static adsorption and desorption tests were carried out to screen the macroporous resins. The desorption experiment was per-formed on the selected D101 resin to optimize the separation process. The effects of resin amount, diameter length ratio, elution flow rate, elution solution concentration and volume were studied. Results:The optimal conditions were as follows:the elution solution was 55% ethanol, the adsorption flow rate was 1 BV·h-1 , the elution flow rate was 5 BV·h-1 , 6 BV 25% ethanol was used to eliminate impurity and 8 BV 55% ethanol was used to elute to obtain the effective fraction. Conclusion: The content of antibacterial effective component is above 65% after purified by D101 resin, indicating that the present method is suitable for large-scale preparation of anti-bacterial effective fraction of Syringae folium.

To compare the characteristics of absorption and pharmacokinetic behavior of ginsenoside Rg1 (Rg1) with ginsenoside Rb1 (Rb1) of panax notoginseng saponins (PNS), bile excretion of both Rg1 and Rb1 were studied after i.v. and i.g. of PNS solution. Plasma protein binding ratios were studied using equilibrium dialysis method, and referred to pharmacokinetic parameters. It shows that (61.48 +/- 18.30)% dose of Rg1 and (3.94 +/- 1.49)% dose of Rb1 were separately excreted into bile 10 hours after i.v. administration (PNS 50 mg x mL(-1)), and (0.91 +/- 0.51)% dose of Rg1 and (0.055 +/- 0.02)% dose of Rb1 were excreted into bile 12 hours after i.g. administration (PNS 1 500 mg x mL(-1)). Plasma protein binding degrees of Rg1 and Rb1 were 6.56% - 12.74% and 80.1% - 89.69%, respectively. Stomach, intestinal and hepatic throughput efficiency (F(S), F1 and F(H)) for Rg1 were 49.85%, 13.05%, 50.56%, respectively, and 25.82%, 4.18%, 65.77% for Rb1. Therefore, poor intestinal absorption is a primary reason for the low bioavailability of both Rg1 and Rb1. Rg1 possesses relatively high bile excretion and low plasma protein binding rate, in contrast, Rb1 possesses low bile excretion and high plasma protein binding rate. Membrane permeability and elimination rate of Rb1 were lower than that of Rg1, meanwhile, longer MRT and bigger AUC could be found for Rb1.
Administration, Oral ; Animals ; Bile ; secretion ; Biological Availability ; Ginsenosides ; metabolism ; pharmacokinetics ; Intestinal Absorption ; Male ; Panax notoginseng ; chemistry ; Plants, Medicinal ; chemistry ; Protein Binding ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage ; isolation & purification ; pharmacokinetics

Water in oil (W/O) microemulsion formulation was developed to enhance intestinal absorption of ginsenoside Rb1 (Rb1) of panax notoginseng (PNS). Effects of W/O microemulsions on pharmacokinetics after intraduodenal administration, membrane fluidity and membrane transport of Rb, were studied in rats, liposomes and parallel artificial membrane permeability assay (PAMPA), respectively. Soybean phospholipids/ethanol (SP/EtOH) was selected as surfactant/cosurfactant, together with PNS 400 mg x mL(-1) solution and various kinds of oils, to prepare 11 W/O microemulsions. Most of the microemulsions can enhance Rb1 intestinal absorption significantly. Besides surfactant/cosurfactant, oil also had an effect on the enhanced absorption and the order of enhancement was as follows: glyceryl laurate approximately = isopropyl myristate > isopropyl palmitate > 2-ethylhexanol palmitate. The effection of absorption enhancement by the long chain glyceride ( > C14) is lower than that by the medium chain glyceride (C8 - C14). Most of W/O microemulsions were found to enhance the membrane fluidity of liposomes to different extents. In PAMPA analysis, efficient permeability coefficient (Pe) of diluted-microemulsion (D-ME) is mostly higher than that of PNS solution, which indicated the components of microemulision can facilitate the membrane permeability of the drug. Meanwhile, linearity correlation between Pe and ratio of relative bioavailability (Fr) was acquired for undiluted microemulison (ME). Therefore, W/O microemulsions can enhance intestinal absorption of Rbr, and this effect may be attiributed to its enhancement on membrane fluidity to a certain degree. PAMPA analysis could be brought into not only the investigation of membrane transport of crude drug, but also conditioned preformulation research (e.g. absorption enhancer etc.).
Animals ; Drug Delivery Systems ; Emulsions ; Ginsenosides ; administration & dosage ; isolation & purification ; pharmacokinetics ; Intestinal Absorption ; Male ; Membrane Fluidity ; Oils ; Panax notoginseng ; chemistry ; Permeability ; Plants, Medicinal ; chemistry ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Surface-Active Agents ; chemistry ; Water

OBJECTIVETo investigate the characteristics of the hepatic pathological and clinical features of patients with hepatitis B virus (HBV) in immune tolerant stage and find the better measure of diagnosing patients chronic infected by HBV in immune tolerant phase.

METHODS135 patients with HBV chronic infection and persistently normal serum alanine aminotransferase (ALT) levels were involved in this study, whose serum HBeAg and HBV DNA were positive. Statistical analysis included the ages, sex, serum levels of HBVDNA, ALT and histological grade. The grades of inflammation and fibrosis were obtained through hepatic biopsy performed on all the patients.

RESULTSMean age in those patients was 22.61 +/- 8.95 years old. All those patients were divided into two groups according to histological grade: low- histological grade group, G < or = 1 and S < or = 1; and high-histological grade group, G > or = 2, S > or = to 2. Levels of histological grade were low in most of patients (99/135). Patients of low-histological grade had no difference in age, sex and family history of HBsAg carriers. Compared with low-normal ALT (ALT less than 30U/L), those with high-normal ALT (ALT > or = 30U/L) had a greater frequencies of high-histological grade. Compared with high HBVDNA (HBVDNA > or = 10(7) copies/ml), those with low HBVDNA (HBVDNA less than 10(7) copies/ml) had a greater frequencies of high-histological grade.

CONCLUSIONLevels of histological grade were low in most of patients with HBV chronic infection, serum HBeAg and HBVDNA positive, persistently normal serum ALT levels, but some of them were high-histological grade. It showed those patients were not all in immune tolerant stage of chronic HBV infection. Examination of ALT and HBVDNA are helpful to evaluate hepatic pathological damage for them.

Adolescent ; Adult ; Alanine Transaminase ; blood ; Child ; Child, Preschool ; DNA, Viral ; blood ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; virology ; Humans ; Immune Tolerance ; Male ; Middle Aged

Acute Disease ; Fatal Outcome ; Gestational Age ; Humans ; Infant, Newborn ; Male ; Respiratory Distress Syndrome, Newborn ; diagnosis ; therapy

Rat aorta media, adventitia and cultured vascular smooth muscle cells (VSMCs) were used in this study to identify the source of nitric oxide (NO) generation from various cell types of vascular tissues and to elucidate the mechanisms involved in NO formation. Treatment of vascular media and VSMCs with lipopolysaccharide (LPS) or cytokines [tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta)] resulted in a dose-dependent increase of NO release. Inducible nitric oxide synthase (iNOS) in the stimulated VSMCs was significantly upregulated as shown by Western blot analysis. Protein kinase C (PKC) inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7) prevented LPS-, TNF-alpha- and IL-1beta-induced NO production, whereas N-(2-guanidinoethyl)-5-isoquinoline-sulfonamide (HA1004), an H7 analogue with little activity towards PKC, had no inhibition effect. The role of PKC in LPS- and cytokine-induced changes on NO formation was confirmed by using another structurally distinct PKC inhibitor chelerythrine. Treatment of VSMCs with protein tyrosine kinase (PTK) inhibitor genistein or tyrphostin AG18 also reduced the NO production evoked by LPS, TNF-alpha or IL-1beta, which was associated with inhibition of iNOS protein expression. In contrast, PKC inhibitor chelerythrine did not affect iNOS expression. These results suggest that PTK mediates LPS- and cytokine-induced NO formation by upregulation of iNOS expression. PKC may be involved in the post-translational modification of iNOS or the regulation of the availability of iNOS substrates and cofactors.
Animals ; Aorta, Thoracic ; cytology ; Cells, Cultured ; Cytokines ; pharmacology ; Interleukin-1beta ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Protein Kinase C ; physiology ; Protein-Tyrosine Kinases ; physiology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To explore the clinical significance of preoperative ultrasound-assisted drawings in location of thyroid micronodule resection .Methods A total of 88 patients ( 137 nodules ) who underwent thyroid micro-nodule resection were enrolled in the prospective randomized controlled study .All patients were randomly divided into two groups:46 patients (68 micronodules) in experimental group and 42 patients (69 micronodules) in control group.Inclusion criteria:the maximum diameter of nodule≤1.0 cm. Preoperative thyroid ultrasound was conducted .Patients in experimental group also underwent ultrasound-assisted location of thyroid micronodule .Results The diagnostic accuracy rate of US was 81.82%(69/88) in all patients .With the help of ultrasound-assisted location , all nodules in experimental group were found quickly and accurately.The resection rate of experimental group was 100%(46/46).Whereas 4 nodules (in 4 patients ) were missed in control group with a resection rate of 90.5% (38/42).The postoperative US examinations after 3 months showed that all nodules were completely removed in experimental group while 4 nodules retained in control group .Conclusions Preoperative ultrasound-assisted drawings in location of thyroid micronodule plays an important role in thyroid nodule resection .It is great value of clinical application .

BACKGROUNDMycoplasma pneumoniae (M. pneumoniae) is one of the common pathogens causing atypical pneumonia. In recent years, resistance to macrolides has become more common, especially in China. Previous studies have confirmed that the mutation at position 2063 in domain V of the 23S rRNA is the most prevalent, followed by the mutation at position 2064. Reported molecular detection methods for the identification of these mutations include direct sequencing, restriction fragment length polymorphism analysis, real-time polymerase chain reaction (PCR) with high-resolution melt analysis, and nested PCR-linked with capillary electrophoresis, etc. The most commonly used method for monitoring resistance-conferring mutations in M. pneumoniae is direct DNA sequencing of PCR or nested PCR products. However, these methods are time-consuming, labor-intensive or need expensive equipments. Therefore the development of rapid and sensitive methods is very important for monitoring the resistance globally.

METHODSIn this study, we reported a fast and cost-effective method for detecting 2063 and/or 2064 macrolide resistant mutations from specimens using a modified allele-specific PCR analysis, and all results were compared with the sequencing data. We also analyzed the clinical courses of these samples to confirm the modified allele-specific PCR results.

RESULTSAmong 97 M. pneumoniae specimens, 88 were found to possess mutations by this method, and all modified allele-specific PCR analysis results were consistent with the sequencing data. The data of the clinical courses of these 97 cases showed that they suffered from severe pneumonia. Erythromycin showed better efficacy on cases from which no macrolide resistance mutation was found on their specimens. However, in some cases from which mutations were detected, erythromycin monotherapy had poor efficacy, and on these patients severe symptoms improved only when azithromycin was added to the treatment.

CONCLUSIONSThe drug-resistant M. pneumoniae is very common in Beijing, China. Our modified allele-specific PCR analysis can identify erythromycin resistant mutations more rapidly from specimens than any other method currently available. Erythromycin is still effective for treating patients infected with the mutation negative M. pneumoniae, but this treatment fails to work on mutant organisms. This method can facilitate clinicians in selecting appropriate therapy within short timescales.

Alleles ; Anti-Bacterial Agents ; pharmacology ; China ; Drug Resistance, Bacterial ; genetics ; Erythromycin ; pharmacology ; Mycoplasma pneumoniae ; drug effects ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo compare the knee osteoarthritis (OA) models in rabbits by different concentrations of papain and provide data for exploring pathogenesis and treatments of this disease.

METHODSSixty New Zealand white rabbits were randomly divided into four groups of 15 each and given injections into the right knee on days 1, 3 and 5 including intra-articular injections of 2%, 5% or 10% (w/v) papain and 0.03 mol/L L-cysteine at the dose of 0.1 ml/kg (experimental groups). The 0.9% NaCl (w/v) with a dose of 0.1 ml/kg were injected intra-articularly into the right knees of rabbits in the control group. The rabbits were sacrificed at 2, 4, 6 weeks respectively after the initiation of papain injection and these OA models were evaluated through recording the width of knee joint, performing the morphological observation and histological evaluation of articular cartilage and synovium.

RESULTSThe degenerative changes were demonstrated in knee joints of rabbit in all experimental groups, such as thinner articular cartilage, fibrillation and destroyed cartilage matrix, and inflammation, proliferation, and degeneration of the synovial tissue. All these changes were much worse with increased concentration and prolonged observation time.

CONCLUSIONDifferent severity of OA are established through intra-articular injections of 2%, 5% or 10% papain and 0.03 mol/L L-cysteine at the dose of 0.1 ml/kg. These models are of the characters of short period and a good reproducibility.

Animals ; Disease Models, Animal ; Male ; Osteoarthritis, Knee ; chemically induced ; pathology ; Papain ; toxicity ; Rabbits

OBJECTIVETo investigate the protein expression of cyclooxygenase 2 (COX-2) and nuclear factor kappa B (NF-kappaB) in gastric mucosa-associated lymphoid tissue (MALT) lymphoma and its clinical significance.

METHODSProtein expression of COX-2 and NF-kappaB in gastric MALT lymphoma were examined by immunohistochemistry of Envision two-step method. The correlations of COX-2 and NF-kappaB expression with Helicobacter pylori (Hp) infection, clinical stage, depth of tumor invasion, tumor size, recurrent rate and treatment were analyzed by univariate, multivariate and Pearson analysis.

RESULTSThe positive expression of COX-2 and NF-kappaB in gastric MALT lymphoma were 48.9%(23/47) and 36.2% (17/47) respectively, and a positive correlation was found between these two factors(r=0.326,P<0.05). Moreover, COX-2 expression was positively correlated with Hp infection,clinical stage, depth of invasion and tumor size (P<0.05). Univariate analysis showed that the overall survival of gastric MALT lymphoma patients with positive COX-2 protein (59.9 months) was shorter than that of patients with negative COX-2 protein (77.8 months), but the difference was not significant (P>0.05). The survival was significantly shorter in gastric MALT lymphoma patients with positive NF-kappaB protein (26 months) than that of patients with negative NF-kappaB protein (123.2 months)(P<0.05). Multivariate Cox regression analysis revealed that clinicopathological stage was independent prognostic factor, and associated with short survival.

CONCLUSIONUp-regulated expression of COX-2 and activation of NF-kappaB are associated with Hp infection in gastric MALT lymphoma, and their protein expression is correlated with the development of tumor and prognosis.

Cyclooxygenase 2 ; metabolism ; Female ; Gastric Mucosa ; metabolism ; microbiology ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Lymphoma, B-Cell, Marginal Zone ; metabolism ; microbiology ; pathology ; Male ; Middle Aged ; NF-kappa B ; metabolism ; Neoplasm Staging ; Prognosis ; Stomach Neoplasms ; metabolism ; microbiology ; pathology