Objective To investigate the effect of LI-cadherin- SiRNA protein on the growth and metastatic potentials of transplanted human hepatocellular carcinoma cell lines (Hep3B) in nude mice.Methods We transfected LI-cadherin- SiRNA to Hep3B cells,Hep3B cell suspension (transfected or control ) was injected subcapsullaryly into the spleen of nude mice,hepatic metastasis was observed by naked eye and immunohistochemistry.In addition,Western-blot was used to detect the level of LI-cadherin in different metastatic site.Results (1) Hep3B was green after successful transfect interference vector under fluorescence inverted microscope,in this study,the transfect rate was 80％.(2) Hep3B liver metastasis model in nude mice was established.The metastasis rates in the empty plasmid carrying group,the control group and the SiRNA transfect group were 50％,60％ and 80％,respectively.The number of metastasis caner nodules in the SiRNA transfect group was 26,significantly higher than other two groups.(3) The level of protein expression for LI-cadherin in the SiRNA transfect group is significantly lower than the control group and the empty plasmid carrying group.Conclusions LI-cadherin is crucial and important for the adhension capability of HCC in its migration.SiRNA transfected LI-cadherin increases the metastasis in a nude mouse model inoculated with human hepatocellular carcinoma cell lines.

Abstract： Objective To evaluate the filtration effects of various nanofiltration systems on intravenous human immunog⁃ lobulin（IVIG）in order to screen the optimal nanofiltration system. Methods Various nanofilters were used for IVIG filtration to determine the best one and then various prefilters were selected to combine with the optimal nanofilter for IVIG filtration to determine the optimal nanofiltration system. Results The tangential flow（cross flow）nanofilter showed better filtering effect than dead end（direct current）nanofilter，and nanofilter C was the best one. The effect of deep filtration prefilter was better than that of absolute filtration prefilter，and prefilter Y1 in series with nanofilter C was the optimal nanofiltration system. Conclusion The optimal nanofiltration system was determined through the effect evaluation of various nanofiltration systems filtering for IVIG.

In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 μm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum
Acetonitriles ; Chromatography, High Pressure Liquid ; Flavonoids ; chemistry ; Kaempferols ; Quercetin ; analogs & derivatives ; Rutin ; Tandem Mass Spectrometry ; Vitaceae ; chemistry

OBJECTIVETo observe the analgesic effects of single-and combined-laser irradiation with low-intensity applied at "Zusanli" (ST 36) in rats, and their relation to degranulation of mast cells.

METHODSSixty-six SD rats were randomly divided into 6 groups: normal control group (Group NC), model control group (Group MC), sham irradiation group (Group SI), 10.6 microm laser irradiation group (Group 10.6 microm LI), 650 nm laser irradiation group (Group 650 nm LI) and combined (10.6 microm + 650 nm) laser irradiation group (Group CLI). Complete Freund's Adjuvant (0.05 mL) was injected into the left ankle joints of all the rats except those in Group NC to cause acute adjuvant-induced arthritis. In treatment, laser irradiation was applied at "Zusanli" (ST 36) for 30 minutes in all the rats except those in Group NC and Group MC. The paw withdrawal latency (PWL) to radian heat was used to compare analgesic effects among the groups. By means of toluidine blue, dyed slices of local tissues of "Zusanli" (ST 36) were used to observe changes of mast cell degranulation before and after laser irradiation.

RESULTSThe pain thresholds to irradiation of the rats in Group 650 nm LI and Group CLI were significantly higher than those in Group MC and Group SI (P < 0.01), and the mast cell degranulation rate in Group 650 nm LI and Group CLI were also significantly higher than that in Group MC and Group SI (P < 0.001). The pain threshold and mast cell degranulation rate in Group 10. 6 microm LI were not significantly different from those in Group MC and Group SI. There was a linear correlation between mast cell degranulation rate and PWL with 0. 737 in coefficient (P < 0.001).

CONCLUSIONSingle 650 nm laser and combined 650 nm + 10.6 microm laser with low intensity irradiated at "Zusanli" (ST 36) in acute adjuvant rats can provide remarkable analgesic effects, and there was a positive correlation between mast cell degranulation rate and analgesic effects, which plays an important part in laser irradiation-induced analgesia.

Acupuncture Analgesia ; methods ; Acute Disease ; Animals ; Arthritis, Experimental ; chemically induced ; physiopathology ; therapy ; Cell Degranulation ; radiation effects ; Combined Modality Therapy ; Freund's Adjuvant ; Low-Level Light Therapy ; methods ; Male ; Mast Cells ; cytology ; physiology ; Pain Measurement ; Pain Threshold ; radiation effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To explore Chongqing migrant workers Suiqian children′s mental health status and related factors .Meth-ods In this study 740 children from grade 7 to grade 9 in 3 schools which specify recruit migrant workers′children ,including the 399 migration children of migrant workers and 341 urban household registration students were involved .They were investigated with symptom rating scale(SCL-90) and self-made general questionnaire .Results There were no significant differences in SCL-90 scores between the migrant workers migration children group and urban household registration students group (P>0 .05) .Conclu-sion Generally speaking ,Chongqing migrant workers Suiqian children′s mental health status is good ,but there are still some prob-lems .We need do some further study on the mode of psychological intervention to maintain their physical and mental health .

The cloned TWIK-related acid-sensitive K(+) channel (TASK-1) is sensitive to the pH changes within physiological pH range (pK~7.4). Recently, the native TASK-1-like channel was suggested to be the main contributor to the background (or leak) K(+) conductance in the motoneurons of the brain stem. Serotonin (5-HT) and variation of pH value in perfused solution could modulate these currents. Here we aimed to examine the properties and modulation of the currents by serotonin or variation of pH value in hypoglossal motoneurons of rats. Transverse slices were prepared from the brainstem of neonatal Sprague-Dawley rats (postnatal days 7-8). Hypoglossal motoneurons were used for the study. The leak K(+) current (TASK-1-like current) and hyperpolarization-activated cationic current (I(h)) were recorded with the whole-cell patch-clamp technique. The results showed that these currents were inhibited by acidified artificial cerebrospinal fluid (ACSF, pH 6.0) and activated by alkalized ACSF (pH 8.5). 5-HT (10 mumol/L) significantly inhibited both leak K(+) current and I(h) with depolarization of membrane potential and the occurrence of oscillation and/or spikes. Bath application of Ketanserine, an antagonist of 5-HT₂ receptor, reversed or reduced the inhibitory effect of acidified solution on leak K(+) current and I(h). The results suggest that 5-HT₂ receptors mediate the effects of acidified media on leak K(+) current and I(h) in hypoglossal motoneurons.
Animals ; Animals, Newborn ; Brain Stem ; cytology ; Hypoglossal Nerve ; cytology ; In Vitro Techniques ; Ion Transport ; Membrane Potentials ; Motor Neurons ; metabolism ; Patch-Clamp Techniques ; Potassium Channels, Tandem Pore Domain ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serotonin ; pharmacology

OBJECTIVETo explore the influence of exogenous putrescine and cadaverine on pro-inflammatory factors in the peripheral blood of rabbits.

METHODSForty ordinary adult New Zealand rabbits were divided into saline, necrotic tissue homogenate (NTH), putrescine, and cadaverine groups according to the random number table, with 10 rabbits in each group. Saline, NTH, 10 g/L putrescine, and 10 g/L cadaverine were respectively peritoneally injected into rabbits of corresponding group in the amount of 1 mL/kg. The blood sample in the volume of 2 mL was collected from the central artery of rabbit ears before injection and at 2, 6, 12, 24, 30, 36, 48, 60 hours post injection (PIH). Contents of TNF-α, IL-1, and IL-6 in the serum were determined with enzyme-linked immunosorbent assay. Data were processed with repeated measurement data analysis of variance and Spearman correlation analysis, and cubic model curve was applied in curve fitting for the contents of inflammatory factors.

RESULTS(1) The serum contents of TNF-α, IL-1, and IL-6 were increased in NTH, putrescine, and cadaverine groups in different degrees at most post injection time points. There was no significant change in the concentrations of the three pro-inflammatory factors in saline group, and they were significantly lower than those of the other three groups at most post injection time points (with F values from 3.49 to 13.58, P values all below 0.05). The serum contents of TNF-α, IL-1, and IL-6 in putrescine group began to increase at PIH 2, 6, and 6, which was similar to the trend of NTH group, but the changes were delayed compared with those of cadaverine group(all at PIH 2). The peak values of TNF-α, IL-1, and IL-6 in putrescine group were respectively (339 ± 36), (518 ± 44), and (265.9 ± 33.5) pg/mL, which were significantly lower than those of cadaverine group [ (476 ± 86), (539 ± 22), and (309.4 ± 27.1) pg/mL], with F values respectively 5.11, 1.90, and 5.56, P values all below 0.05. (2) The period of time in which contents of TNF-α, IL-1, and IL-6 began to increase (PIH 3-4) and the peaking time of the three pro-inflammatory cytokines (PIH 18-30) in putrescine group appeared later than those of cadaverine group (PIH 2 and 12-30). The duration of peaking time of the three pro-inflammatory cytokines in putrescine group was shorter than that of cadaverine group (PIH 18-30 vs. PIH 12-30). The increasing period and the duration of peaking time of TNF-α, IL-1, and IL-6 in putrescine group were close to those of NTH group (PIH 3-5 and 18-30). The correlation coefficient test analysis showed that the trends of changes in contents of three pro-inflammatory cytokines in putrescine group were significantly correlated with those of NTH group (r(TNF-α) = 0.933, P < 0.01; r(IL-1) = 0.967, P < 0.01; r(IL-6) = 0.950, P < 0.01). The obvious correlation between cadaverine group and NTH group was only found in the contents of IL-1 and IL-6 (r(IL-1) = 0.913, P < 0.01; r(IL-6) = 0.883, P < 0.05).

CONCLUSIONSBoth exogenous putrescine and cadaverine can cause inflammatory reaction in rabbits. The trend of the inflammatory reaction induced by putrescine was similar with that by NTH, suggesting that putrescine may play a leading role in the inflammatory reaction induced by necrotic tissue decomposition.

Animals ; Cadaverine ; adverse effects ; Inflammation ; blood ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Necrosis ; blood ; Putrescine ; adverse effects ; Rabbits ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo evaluate the application of fluorescence in-situ hybridization (FISH) in detection of gene translocation in paraffin-embedded tissue samples of synovial sarcoma.

METHODSInterphase FISH was carried out in paraffin-embedded tissue of 42 cases of synovial sarcoma and 9 cases of non-synovial sarcoma, using a LSI SYT (18q11.2) dual color break-apart probe. In all of the cases studied, the gene fusion product SYT-SSX was also analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSPositive signals were detected in 37 cases (88.1%) of synovial sarcoma by FISH, as compared with 35 cases (83.8%) by RT-PCR and 39 cases (92.9%) by both techniques. Of the 39 positive cases, 33 cases (78.5%) revealed SYT gene translocation.

CONCLUSIONSFISH may serve as an adjunctive diagnostic tool in problematic cases of synovial sarcoma and can be applied in paraffin-embedded tissue samples. As compared with RT-PCR, FISH is also sensitive and reliable. The methodology is less labor intensive and time consuming. FISH has great potential in molecular diagnosis of soft tissue tumors.

Adolescent ; Adult ; Biomarkers, Tumor ; genetics ; Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Head and Neck Neoplasms ; genetics ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Lower Extremity ; pathology ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Paraffin Embedding ; Reverse Transcriptase Polymerase Chain Reaction ; Sarcoma, Synovial ; genetics ; metabolism ; Soft Tissue Neoplasms ; genetics ; metabolism ; Young Adult

Twenty five new beta-aminoalcohols containing nabumetone moiety were prepared via the reduction of potassium borohydride with a convenient and efficient procedure, starting from beta-aminoketones that have been synthesized by our group. Their chemical structures were determined by IR, MS, 1H NMR, 13C NMR, HR-MS and antidiabetic activities were screened in vitro. Preliminary results revealed that the antidiabetic activity of most beta-aminoalcohols were better than that of the corresponding beta-aminoketones. Although most compounds showed weak antidiabetic activity, the alpha-glucosidase inhibitory activity of compounds 5hd(1) and 5id(2) reached 74.37% and 90.15%, respectively, which were superior to the positive control. The relative peroxisome proliferator-activated receptor response element (PPRE) activity of five compounds were more than 60%, among them compound 5ca possessed the highest activity (112.59%). As lead molecules of antidiabetic agents, compounds 5hd(1), 5id(2) and 5ca deserve further study.
Amino Alcohols ; chemical synthesis ; chemistry ; pharmacology ; Butanones ; chemical synthesis ; chemistry ; pharmacology ; Cyclooxygenase 2 Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Glycoside Hydrolase Inhibitors ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Peroxisome Proliferator-Activated Receptors ; agonists ; metabolism ; Response Elements ; alpha-Glucosidases ; metabolism

A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.
Acetonitriles ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Formates ; chemistry ; Tandem Mass Spectrometry ; methods ; Vitaceae ; chemistry ; Water ; chemistry