We hypothesized that the concentrated urea and NaCl solutions may opened the blood-brain barrier to the horseradish peroxidase and trypan blue-albumin complex by shrinking barrier cells and opening up spaces between them. The experiments were carried out on 30 healthy, adult rats. Two experimental groups were used. First, intracarotid perfusion of anesthetized rats were prepared by exposing and catheterizing the left common carotid artery. A test solution of 3.4 M, 3.0 Osm urea and 0.87 M, 1.6 Osm NaCl, was perfused manually for 30 sec. in a cranial direction so as to expel the blood from the pial arterioles of the exposed brain. The pressure, which was not measured, varied between what was required which to expel blood from both arterioles and venules. Five milliliters of the test solution usually were used. Horseradish peroxidase and trypan blue was injected intravenously or through the carotid artery after perfusion. Threshold of barrier damage due to the intracarotid substance was defined as the lowest osmotality which produced obvious blue staining of the brain both on surface observation and coronal section. The effect of a substance was defined as reversible if a threshold concentration did not produce blue staining when the dye was injected 30 min following perfusion. Second, we applied a concentrated solution of 3.0 Osm urea and 1.6 Osm NaCl to the pia-arachnoid of the cerebral cortex, to study the barrier to the intravascular horseradish peroxidase and trypan blue-albumin complex. Hyperosmotic solution of 3.0 Osm urea and 1.6 Osm NaCl, either infused into one internal corotid artery or applied topically to the pia-arachnoid surface of the brain of rats, results in the opening of endo thelial tight junction through which horseradish peroxidase passes from blood to the basal menbrane and astrocytes and neurons. The evidence for this opening of the blood-brain barrier to protein is the entry of peroxidase into the neurons. It was postulated that sufficiently high concentrations of electrolytes or relatively lipid-insoluble non-electrolytes such as urea, osmotically pulled water from the cerebral endothelial cells resulting in their shrinkage. The shrinkage, in turn, was believed to open the tight junction between continuous endothelial cells so that the dyeprotein complex could pass through the junction from blood to neurons. The present study shows that these tight junctions, the sites of the barrier to neuron movement of protein, are indeed opened by the osmotic action of urea or NaCl.

Objective To establish an HPLC method for simultaneous contents determination of paeoniflorin, protocatechuic acid and chlorogenic acid in Penyanjing Capsule. Methods The chromatographic separation was performed on a SHIMADZU VP-ODS-C18 column (4.6 mm × 250 mm, 5 μm); the column temperature was maintained at 30 ℃; A gradient elution of acetonitrile-0.15% phosphoric acid aqueous solution was adopted at the flow rate of 1.0 mL/min; The UV detection wavelengths were at 231, 255, and 326 nm; The injection volume was 20 μL. Results Paeoniflorin, protocatechuic acid and chlorogenic acid could be separated efficiently with this condition. The regression equation was Y=1546.4128X+127.0756 (r=0.9999), Y=2925.8468X+2204.1076 (r=0.9998), Y=893.9045X-261.7315 (r=0.9994), respectively. Paeoniflorin, protocatechuic acid and chlorogenic acid were linear in the range of 150.054–1254.50 ng, 162.33–1352.75 ng, 11.43–95.25 ng, and the average recoveries were 97.60%, 102.09% and 98.52%. Conclusion The method is simple, convenient and accurate, which can be used to the quality control of Penyanjing Capsule.

Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

To determine the potential of the high mobility group box-1 protein 1 (HMGB1) to activate Toll-like receptor 4 (TLR4) signaling in hepatic stellate cells (HSCs) and investigate the subsequent transition of HSC towards the inflammatory phenotype. Three immortalized mouse HSC cell lines, wild-type (JS1), TLR4-/- (JS2) and MyD88-/- (JS3), were subcultured in plates and divided into groups of normal control (untreated), postive control (lipopolysaccaride, LPS treatment), and experimental (HMGB1 treatment). All groups were transfected with luciferase reporter plasmids carrying responsive elements for either the nuclear factor-kappa B (NF-kB) or activator protein-1 AP-1 transcription factors. Following stimulation with normal saline, LPS (100 ng/mL) or HMGB1 (100 ng/mL), the activation of NF-kB or AP-1 was detected by a dual-luciferase reporter assay system. The induction of monocyte chemotactic protein-1 (MCP-1) transcription was determined by measuring the mRNA levels using real time quantitative reverse transcription PCR (qRT-PCR). The secreted protein levels of MCP-1 were determined by enzyme-linked immunosorbent assay (ELISA) of the culture supernatants. Activation of NF-kB- and AP-1-responsive reporters was significantly up-regulated in JS1 cells treated with HMGB1 or LPS, and the activation was coincident with markedly up-regulated transcription and secretion of MCP-1. However, HMGB1 and LPS treatment produced no responsive of the NF-kB and AP-1 reporters, and no increase in expression or secretion of MCP-1, in JS2 or JS3 cells. As an endogeous ligand of TLR4, HMGB1 may activate TLR4 signaling and the TLR4-mediated inflammatory response of HSC.
Animals ; Cell Line ; Chemokine CCL2 ; genetics ; metabolism ; Gene Knockout Techniques ; HMGB1 Protein ; pharmacology ; Hepatic Stellate Cells ; drug effects ; metabolism ; Lipopolysaccharides ; pharmacology ; Mice ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; genetics ; metabolism ; Transcription Factor AP-1 ; genetics ; metabolism ; Transfection ; Up-Regulation ; drug effects

OBJECTIVE: To detect pathogenic variant in a juvenile with severe type Cornelia de Lange syndrome (CdLS). METHODS: A 12-year-old female presented with comprehensive developmental retardation and deformity of lower limbs. Genomic DNA was extracted from peripheral blood sample of the patient. Whole exome sequencing was performed to identify pathogenic variants. Putative variant was verified by Sanger sequencing. The impact of variants was predicted and validated by bioinformatic analysis. RESULTS: A de novo missense variant, c.1507A>G (p. Lys503Glu), was found in the NIPBL gene of the proband. The variant was unreported previously and predicted to be pathogenic by PolyPhen-2, MutationTaster and SIFT. Using HomoloGene system, the 503 loci in the NIPBL protein are highly conserved. The change of amino acid (Glu), locating in 503 locus, was found to cause the Neuromodulin_N superfamily domain destroyed, resulting in severe damage to the function of NIPBL protein. CONCLUSION The de novo missense variant c.1507A>G (p. Lys503Glu) of the NIPBL gene probably underlies the disease in this patient.
Cell Cycle Proteins ; genetics ; Child ; De Lange Syndrome ; genetics ; Developmental Disabilities ; genetics ; Female ; Humans ; Mutation, Missense ; Phenotype

Objective:To study the effect of Sanjie Xiaoliu Recipe on transplanted tumors in breast cancer mice through in vivo experiments, in order to explore the efficacy and mechanism of Sanjie Xiaoliu Recipe on breast cancer patients. Methods:45 BaL B/c female mice were selected to establish the transplanted tumor model of breast cancer. All the transplanted tumor models of breast cancer mice were randomly divided into five groups: the control group (intragastric administration of normal saline), the low, medium and high dose group of Sanjie Xiaoliu Recipe (intragastric administration of different doses of Sanjie Xiaoliu Decoction), and the paclitaxel group (intraperitoneal injection of paclitaxel). After 24 days of continuous administration, the diet and activities of mice were observed; the body weight, weight and volume changes of transplanted tumor mice were recorded, and the tumor inhibition rate was calculated. Western blot was used to detect the expression of epithelial mesenchymal transformation related proteins (E-cadherin, N-cadherin, Vimentin) in the transplanted tumor tissues of mice in each group.Results:(1) The food intake and activity status of mice treated with Sanjie Xiaoliu Recipe were less affected by the transplanted tumor of breast cancer. (2) The volume and weight of transplanted tumor in the treat groups were smaller than those in the control group (all P<0.01), and the volume of transplanted tumor in the middle dose group of Sanjie Xiaoliu Recipe was smaller than that in the low dose group ( P<0.05). The tumor inhibition rates among the treatment groups were: Sanjie Xiaoliu Recipe medium dose group 52.4%, paclitaxel group 40.3%, Sanjie Xiaoliu Recipe low dose group 39.5%, Sanjie Xiaoliu Recipe high dose group 34.1%. (3) The results of Western blot showed that the expression level of E-cadherin in the transplanted tumor tissue of the treat groups was higher than that in the control group, and the expression levels of N-cadherin and Vimentin were lower than that in the control group, with statistically significant difference (all P<0.05). Conclusions:The Sanjie Xiaoliu Recipe can improve the weakness and reduced consumption of breast cancer mice, which can inhibit the tumor mass growth in mice to a certain extent. Its mechanism may be that Sanjie Xiaoliu Recipe can inhibit the epithelial mesenchymal transformation of breast cancer and the invasion and metastasis of breast cancer.

OBJECTIVETo evaluate the clinical value of double contrast-enhanced ultrasonography using oral and intravenous contrast agents in preoperative staging of gastric cancer.

METHODSSixty-two patients with biopsy-proven gastric cancer were enrolled into this study, and were examined by double contrast-enhanced gastric ultrasonography preoperatively. The results were compared with postoperative pathologic findings.

RESULTSThe accuracy of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 72.9% (T1: 66.7%, T2: 60.0%, T3: 76.9%, T4: 71.4%) and 88.1% (T1: 66.7%, T2: 80.0%, T3: 89.7%, T4: 100%), respectively, with a statistically significant difference between the two methods (P = 0.036). The sensitivity, specificity, accuracy and Youden index of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in assessment of lymph node metastasis were 74.5%, 66.7%, 72.9%, and 0.41 versus 89.4%, 75.0%, 86.4%, 0.76, respectively. No significant difference in the accuracy of assessment for lymph node metastasis was observed (P > 0.05).

CONCLUSIONDouble contrast-enhanced ultrasonography is useful for preoperative staging of gastric cancer, especially for T staging.

Adult ; Aged ; Contrast Media ; Endosonography ; methods ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Preoperative Care ; Prospective Studies ; Stomach Neoplasms ; diagnostic imaging ; pathology ; surgery ; Sulfur Hexafluoride

OBJECTIVETo investigate the effects of cordyceps acid and cordycepin on the inflammatory phenotype and fibrogenic property of hepatic stellate cells (HSCs).

METHODSAn immortalized mouse HSC line (JS1) was stimulated with lippolysaccharide (LPS; 100 ng/ml) to induce an inflammatory response with or without co-administration of cordyceps acid or cordycepin in various concentrations (10, 50, or 200 mumol/L). Effects of the treatments on the chemokine monocyte chemotactic protein-1 (MCP-1) mRNA expression in the cells and the protein secretion in the cell culture supernatants were determined by reverse transcription and real-time quantitative PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. In addition, JS1 cells were treated with transforming growth factor-b1 (TGFb1; 10 ng/ml) to induce a fibrogenic response with or without co-administration of cordyceps acid or cordycepin in various concentrations (10, 50, or 200 mumol/L). Effects on the expression of fibrogenic proteins including collagen type I and a-smooth muscle actin (a-SMA), were investigated by Western blot.

RESULTSHigh-concentration (200 mumol/L) treatments of both cordyceps acid and cordycepin significantly inhibited the LPS-induced up-regulation of MCP-1 transcription and secretion (mRNA: 2.07 +/- 0.29 vs. 3.35 +/- 0.26, t = 15.90 and 1.15 +/- 0.23 vs. 4.17 +/- 0.61, t = 8.93; protein: 1.88 +/- 0.06 vs. 2.33 +/- 0.06, t = 10.39 and 1.47 +/- 0.25 vs. 1.97 +/- 0.04, t = 4.60; all P less than 0.05). All concentrations of cordyceps acid and cordycepin inhibited the TGFb1-induced up-regulation of collagen type I and a-SMA protein expression. However, the effects were more robust with the 200 mumol/L concentrations (P less than 0.05).

CONCLUSIONCordyceps acid and cordycepin ameliorate the LPS-induced inflammatory phenotype and TGFb1-induced fibrogenic response of cultured HSCs. These effects may contribute significantly to the drugs' therapeutic mechanisms to inhibit and resolve liver fibrosis.

Animals ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Cordyceps ; Hepatic Stellate Cells ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Up-Regulation ; drug effects

OBJECTIVETo evaluate the accuracy and utility of ultrasound-guided core needle biopsy (CNB) in the diagnoses of breast lesions.

METHODSThe clinical data of 2152 consecutive breast lesions examined by CNB were reviewed. The histological agreement between core pathology and subsequent excision pathology was studied. The benign diseases without repeat biopsy were followed up.

RESULTSThere were 1461 cancers in final diagnosis among 2152 breast lesions, 1339 cancers were diagnosed by CNB. The false-negative rate of CNB was 3.5% (51/1461), and the underestimation rate was 4.9% (71/1461). In the repeat biopsy, carcinoma was found in 17 (50.0%) of 34 atypical ductal hyperplasia lesions and 25 (46.3%) of 54 papillary lesions. In 1461 cancers, the false-negative rate of ultrasound-guided CNB (2.1%, 22/1068) was significantly lower than that of free-hand-guided CNB (7.4%, 29/393) (P < 0.05). The false-negative rate of two special doctors for CNB (1.2%, 8/681) was significantly lower than that of other doctors (5.5%, 43/780) (P < 0.05). In 738 of benign lesions, 417 cases were excised and 50 malignant lesions were found, 205 cases were followed up by 2 - 29 months (median, 10.2 months), and one malignant lesion was found.

CONCLUSIONSUltrasound-guided core needle biopsy with histopathological assessment is an accurate method in diagnosis of breast lesions. Excisional biopsy is required to the high-risk lesions in CNB.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biopsy, Needle ; instrumentation ; methods ; Breast ; pathology ; Breast Neoplasms ; diagnosis ; pathology ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Reproducibility of Results ; Retrospective Studies ; Sensitivity and Specificity ; Ultrasonography, Mammary