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 .

In order to find highly active antidiabetic lead compound, sixteen 4-aminobenzoic acid derivatives were designed and synthesized directly through Mannich reaction in the solution of ethanol at 15-35 degrees C with facile method, mild reaction condition and high yield (45%-90%). Fifteen of them are new compounds. Their structures were confirmed by 1H NMR, 13C NMR, IR, ESI-MS and HR-MS. Alpha-glucosidase inhibitory activity of these compounds indicated that most of these compounds possess the activity with the order: 2c > 2b > 2h > 1a > 1f. The structure-activity relationship of these 4-aminobenzoic acid derivatives was also discussed.
4-Aminobenzoic Acid ; chemical synthesis ; pharmacology ; Drug Design ; Glycoside Hydrolase Inhibitors ; Hypoglycemic Agents ; chemical synthesis ; pharmacology ; Mannich Bases ; chemistry ; Molecular Structure ; Structure-Activity Relationship ; alpha-Glucosidases ; metabolism ; para-Aminobenzoates

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

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

OBJECTlVE To investigate the antidepressant effect and reIated mechanism of the totaI fIavonoids extract parts( Iicorice fIavonoids,LF)from Glycyrrhiza uralensisFisch. cuItivated IocaIIy in Ningxia. METHODS Forced swimming test( FST)and taiI suspension test( TST)were adopted to study the antidepressant pharmacoIogicaI effect in the acute stress-induced depression modeI in mice. The Km mice were intragastricaIIy administered with LF(5,30 and 180 mg·kg-1 )once daiIy,for 21 con-secutive days. One hour after the first,seventh and Iast administrations,the mice were submitted to FST by recording the immobiIity period within the Iast 4 min of the totaI 6 min in both tests and the resuIts were expressed as decrease in immobiIity period with respect to vehicIe controI. In TST,the other group of Km mice was used to evaIuate the antidepressant effect in same protocoI. In the antagonism of reserpine-induced symptoms test( ART),ICR mice were administered intragastricaIIy with LF( 50,150 and 400 mg·kg-1 )once daiIy for 7 consecutive days. One hour after the Iast administration,the mice received reserpine(4 mg·kg-1 ,ip),and ptosis or akinesia was measured 1 h after reserpine injection whiIe rectaI temperature was measured 4 h after the reserpine injection respectiveIy. The same protocoI was adopted in yohimbine toxicity potentiation test(YTT)as in ART. Thirty minutes fter the Iast adminis-tration,the mice received the threshoId IethaI dosage of yohimbine(30 mg·kg-1 ,sc)respectiveIy,and the death number of the mice was caIcuIated in 24 h after the yohimbine administration. In the 5-hydroxy-L-tryptophan(5-HTP)induced head-twitches test(HTT)in mice,after being administered intragastricaIIy with LF(50,150 and 400 mg·kg-1 )once daiIy for 7 consecutive days,the mice received pargiIine (100 mg·kg-1 ,ip)the next day,and 30 min Iater,5-HTP(10 mg·kg-1 ,ip)was intraperitoneaIIy injec-ted to induced the head twitch respectiveIy,and the times of head twitch in a 30 min period after 5-HTP treatment were observed at 6 time points. After HTT,the mice were sacrificed quickIy,and the mono-amine oxidase(mAO)activity in the brain cortex,hippocampus and thaIamus was examined to evaIuate the antidepressant effect of fIavonoids with mAO inhibition. RESULTS Compared with the vehicIe controI,LF significantIy decreased the immobiIity period in both FST and TST(P﹤0.05). LF(50,150 and 400 mg·kg-1 )antagonized the ptosis and akinesia symptoms respectiveIy in 1 h after reserpine administration( P ﹤ 0. 05 ), but faiIed to antagonize hypothermia produced 4 h after reserpine administration. AIso,at the same dosage,LF did not synergeticaIIy produce the enhancement of death by subcutaneous injection of yohimbine at the threshoId IethaI dosage. LF(150 and 400 mg·kg-1 )couId significantIy and synergeticaIIy increase 5-HTP induced head-twitches response(P﹤0.05),but LF couId not promote mAO activity in the cortex,hippocampus and thaIamus at the same dosage. CONCLUSlON LF exerts antidepressant-Iike effect on the modeI of acute despair test. The mechanism might be reIated to direct enhancement of the serotonergic neuraI function in the brain.

Objective To explore the changes of cardiovascular system of military personnel during highly intensive training.Methods One hundred and seventy officers and soldiers were as research subjects,including 100 individuals in high-intensity group and 70 in control group.The levels of serum cortisol (COR),high sensitivity C-reactive protein (hs-CRP),cardiacspecific Troponin T (cTnT) and human heart fatty acid binding protein (H-FABP) were measured when military training ended.All subjects were tested with fatigue assessment instrument (FAI).The data of blood pressure and electrocardiogram were collected from 40 individuals randomly selected from high-intensity group and 36 from control group before and after training for analyzing the blood pressure,arrhythmia and heart rate variability (HRV).Results The levels of COR (indicator related to stress) and hs-CRP (indicator related to inflammation) were significantly higher in high intensity group than in control group (P＜0.05).Highly intensive training can lead to the emergence of myocardial micro-injury,the levels of cTnT and H-FABP were obviously higher than those in control group (P＜0.01),and the mean blood pressure and the severity of fatigue status were significantly higher than those in control group (P＜0.05).The incidence of severe ventricular arrhythmia was lower in both groups (P=1.000).The average heart rate,total heart beats,total number of atrial premature beat,total number of ventricular premature beat,the incidence of sinus arrhythmia and intermittent second degree type Ⅰ atrioventricular block were significantly higher in high intensity group than in control group (P＜0.05).The HRV of high intensity group was obviously decreased (P＜0.01).Conclusion Highly intensive training may induce the military personnel into the state of acute stress and inflammation,which may lead to myocardial injury,increase severity of fatigue status,accompanied with the rise of blood pressure,low HRV and increased incidence of various arrhythmias.