Objective To establish a serum protein fingerprinting technique with a pattern matching algorithm to distinguish Dukes A stage from colorectal cancer of Dukes B,C,D stage. Methods Serum samples were collected from both reserved group and test group, each of the groups comprised 10 patients of colorectal cancer in Dukes A stage and 68 patients in Dukes B,C,D stage. The sera of the reserved group was combined with the surface of the IMAC3 proteinchip. Then the data of SELDI TOF MS was read and analyzed by BioMarker Wizard software and BioMarker Pattern software to get a classificatory pedigree tree, which is a standard configuration that can distinguish the sera of colorectal cancer patients of Dukes A from colorectal cancer patients of Dukes B,C,D, and the standard was confirmed by double blind test in the test group. Results At the M/Z values of 8 320 Da, 8 604Da, 8 867Da and 15 872Da, the protein contents in reserved group are obviously different between the two classes of patients. The accuracy of classification was 97 4%(76/78), corresponding sensitivity was 100%(10/10) and corresponding specificity was 97 1%(66/68). By double blind examination in test group, the corresponding accuracy was 100%(10/10), the corresponding sensitivity was 94 1%(64/68) and the corresponding specificity was 94 1%(64/68). Conclusion Colorectal cancer of Dukes A can be quickly and exactly diagnosed by this method with high sensitivity and specificity. That will be widely used in clinical application

Objective To explore the serum proteomics characteristics of early carcerization of post-hepatitic cirrhosis.MethodsThe serum protein profiles were detected of 62 patients with post-hepatitic cirrhosis and 100 patients with hepatic carcinoma in Ⅰ-Ⅱ stage (T1-2N0M0) and cirrhosis by surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) IMAC30-Cu2+ ProteinChip array (Ciphergen Biosystems Inc.,USA),the distinct proteins were analyzed and a classification tree model was established by using Biomarker Patterns software.The diagnostic efficacy of the model was blindly tested using the serum protein from both groups.ResultsThe serum protein profiles of post-hepatitic cirrhosis and hepatic carcinoma complicating with cirrhosis were analyzed and a classification tree model was established including 4 distinct proteins with different M/Z.In the training mode,the accuracy was 99.4% (161/162) on differential diagnosis to differentiate post-hepatitic cirrhosis and hepatic carcinoma complicating with cirrhosis,the sensitivity and specificity were 99.0% (99/100) and 100.0% (62/62),respectively,on the diagnosis of hepatic carcinoma complicating with cirrhosis.In the testing mode,the accuracy of differential diagnosis was 94.4% (153/162),the sensitivity and specificity were 95.0% (95/100) and 93.5% (58/62) on the diagnosis of hepatic carcinoma complicating with cirrhosis.ConclusionSELDI-TOF-MS ProteinChip technique is of convenient and rapid,and with high sensitivity and specificity on the diagnosis of early carcerization of post-hepatitic cirrhosis.

A high performance liquid chromatography-electrospray ionization mass spectrometry- charged aerosol detection ( HPLC-MS-CAD) method was established for the simultaneous quantitative analysis of four Lignans in Magnoliae Flos extract. The components were separated on a YMC-Pack ODS-A column (250 mm× 4. 6 mm, 5 μm) by gradient elution with methanol and water as the mobile phase at aflow rate of 1. 0 mL/min. Then the elution solution was routed into MS equipment at a flow rate of 0. 3 mL/min and CAD detector at a flow rate of 0. 7 mL/min by a split ratio of 3:7 for the further detection. The column temperature was 25 ℃ and the detection wavelength was 278 nm. A method was developed for the quantitative analysis of muti-components by single maker ( QAMS) to determine pinoresinol dimethylether, magnoli, 1irioresinol B dimethylethe and epi-magnoli A . Magnoli was selected as internal standard and the relative correction factors ( RCF) of the four Lignans were calculated. The contents of the four Lignans in Magnoliae Flos extract were determined by both external standard method and QAMS. The QAMS method was evaluated by comparison of its assay result and that of external standard method. Under the selected chromatographic condition, the limits of detection of pinoresinol dimethylether, magnoli, lirioresinol B dimethylethe and epi-magnoli A were 0. 34, 0. 55, 0. 50 and 0. 58 mg/L, respectively, while the linear range were within 6. 8-270 mg/L, 11-546 mg/L, 2. 0-101 mg/L and 2. 3-116 mg/L. The recoveries ( n=9 ) were 98. 2%-99. 5%, and the correlation coefficient were 0 . 9995-0 . 9998 . No significant differences were found between the quantitative results of external standard method and QAMS method. The developed method is accurate, feasible, and can be used for quality evaluation of Magnoliae Flos .

[Abstract ] Objective Evidence from previous studies indicated that over-activation of C3a/C3aR axis existed in the renal tubular epithelial cells of patients with renal diseases including diabetic nephropathy .However , the pathological significance of over-ac-tivating C3a/C3aR axis still remains to be elucidated .In this study, we constructed a renal tubular epithelial cell line over-expressing C3a in a secretory manner in order to provide a cell model to investigate the pathological significance of over -activating C3a/C3aR axis under various pathological scenes . Methods We designed a synthesized C3a secretory expression unit and cloned it into the multi-clonal site of lentivirus expression vector pLenti 6.3-MCS-IRES2-EGFP.After identification by sequencing , recombinant lentivirus was packaged by using pLenti 6.3-C3a-IRES2-EGFP and packaging plas-mid in 293T cells.Then, the recombinant lentivirus was used to in-fect HK2, a cell line of human renal tubular epithelial cells .After screening in medium with blasticidin , blasticidin resistant cell clones were obtained .Real-time PCR and ELISA method were applied to analyze the expression and secretion of stable transfected cells cloned C3a and identify renal tubular epithelial cell lines with stable over-activating C3a. Results ①C3a secretory expression unit was suc-cessfully synthesized and correctly cloned into the multi-clonal site of pLenti6.3-IRES2-EGFP; ②C3a secrectory expression recombi-nant lentivirus LV-C3a was successfully packaged with a high titer of 5 ×108/mL;③HK2 Cell clones resistant for blasticidin were ob-tained;according to the analysis of Real-time PCR and ELISA, the C3a mRNA level in HK2-C3a cell lines was significantly higher than that of HK2 cells(1.0 ±0.5 vs 1321.0 ±18.0, P<0.01) and the secreted C3a level increased significantly ([0.3 ±0.2]ng/mL vs [249.0 ±37.0] ng/mL, P<0.01). Conclusion The present study successfully constructed C 3a secretory expression vector pLenti6.3-C3a-IRES2-EGFP and C3a over-expression renal tubular epithelial cell line HK 2-C3a, which is very useful in further study of the function and significance of C 3a/C3aR axis not only in renal tubular epithelial cells but also in other cell types .

Objective Our previous study showed that the expression of C 3aR was increased in renal tubular epithelial cells (RTEC).However, the role of C3aR in diabetic nephropathy remains unknown, and so does the exact physiological and pathological signifi-cance of C3aR in relevant renal tissues.In the present study, we investigated the physiological and pathological significance of C3aR in RTECs using a lentivirus expression vector and anRTEC strain overexpressing C3aR. Methods Based on the sequence of C3aR mRNA, the human C3aR gene was synthesized and cloned into the multi-clonal site of the lentivirus expression vector pLenti6.3-MCS-IRES2-EGFP to construct a C3aR expression vector pLenti6.3-C3aR-IRES2-EGFP.After identification by sequencing, the recombinant lentivirus expressing C3aR was packaged by cotransfecting293T cellswith the recombinant lentivirusexpression vector and packaged plasmid .Then,the recombinant lentivirus was used to infect the human RTECline HK 2.After screening in the medium with blasticidin, blasticidinr-esistant cell clones were obtained, followed by identification ofthe human RTECline stably overexpressing C 3aR by real-time PCR and immunochemical staining. Results TheC3aR expression vector pLenti6.3-C3aR-IRES2-EGFP was successfully constructed and the sequence was proved to be correct. C3aR expression recombinant lentivirus was successfully packaged with a titer of 5×108/mL.Blasticidin-resistant HK2 cell clones were ob-tained and the expression of HK2-C3aR mRNA was significantly higherin the HK2-C3aR cells than in the non-transfected HK2 cells (2.33± 0.45 sv 1.00±0.09, P<0.05). Conclusion We successfully constructed a C3aR expression lentivirus vector pLenti6.3-C3aR-IRES 2-EGFP and a C3aR overexpression renal tubular epithelial cell strain HK2-C3aR, which is very contributive to further studies of the roles of C3aR in renal tubular epithelial cells and other types of cells.

Objective To study the effects of environmental exposure to rare earth elements REEs on intelligence of children. Methods Intelligence quotient IQ were examined with Drawing a Man Test in 464 children aged 7-10 years living in RE ore containing area and the control area in Xunwu country Jiangxi province China. Fifteen kinds of REEs in 112 blood samples 69 samples from RE ore area and 43 samples from the control area  were detected by inductively coupled plasma source mass spectrometry ICP-MS. Results All 15 kinds of REEs were detected in each sample the content of blood REEs of the children in the rare earth area 2.18?1.08 ng/g was 1.73 times of that of the children in the control area 1.26?0.35 ng/g the difference was significant P

AlM:To evaluate the efficacy of combined pranoprofen eye drops and artificial tears on the treatment of mild to moderate dry eye syndrome after trabbeculectomy.METHODS:This prospective case control study included 63 cases (63 eyes) of patients with mild to moderate dry eye syndrome after trabbeculectomy in our hospital from November 2013 to June 2013. All subjects were randomly divided into two groups. Observation group was treated with combined pranoprofen eye drops and artificial tears and control group received simple artificial tears marking the eyes at 1, 2, 4wk. The patient's symptoms, signs, BUT, S▏t, and FL were observed before treatment and 1, 2, 4wk after treatment.RESULTS: After 2wk, the symptoms of observation group were improved, there was statistically significant difference (P<0. 05). FL difference of each group was statistically significant ( P<0. 05 ); After 4wk, symptoms and signs were improved. There was statistically significant difference ( P < 0. 05 ). The BUT of the observation group and corneal FL scores of two groups showed significant differences (P<0. 05). CONCLUSlON: Artificial tears joint pranoprofen eye drops has good curative effect in the treatment of mild to moderate dry eye syndrome after trabbeculectomy.

The effect of amygdalin joint hydroxysafflor yellow A (HSYA) on the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and the possible mechanism were studied and explored. Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary endplate chondrocytes. After identification, they were divided into normal group, induced group, amygdalin group, HSYA group and combined group. CCK-8 kit was adopted to detect the proliferation of the endplate chondrocytes. FCM was measured to detect the apoptosis. Real-time PCR method was adopted to observe the mRNA expression of Aggrecan, Col 2 alpha1, Col 10 alpha1, MMP-13 and the inflammatory cytokines IL-1beta. The protein expression of Col II, Col X was tested through immunofluorescence. Compared with the normal group, the proliferation of the endplate chondrocytes decreased while the apoptosis increased (P < 0.05). With down regulation of the mRNA expressions of Aggrecan, Col 2 alpha1 and up regulation of the mRNA expressions of Col 10 alpha1, MMP-13, IL-1beta (P < 0.05), the protein expression of Col II decreased while the protein expression of Col X increased. Compared with the induced group, amygdalin group, HSYA group, the combined group could inhibit the apoptosis and promote the proliferation (P < 0.05). They could increase the mRNA expressions of Aggrecan and Col 2 alpha1 while decrease the mRNA expressions of Col 10 alpha1, MMP-13 and IL-1beta (P < 0.05). They could also enhance the protein expression of Col II while reduce the protein expression of Col X. The effect of the combined group was significantly better than that of amygdalin and HSYA. Amygdalin joint HSYA could inhibit the degeneration of the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and better than the single use of amygdalin or HSYA.
Amygdalin ; pharmacology ; Animals ; Apoptosis ; Cells, Cultured ; Chalcone ; analogs & derivatives ; pharmacology ; Chondrocytes ; drug effects ; Collagen ; metabolism ; Drug Synergism ; Interleukin-1beta ; Intervertebral Disc ; cytology ; Quinones ; pharmacology ; Rats

Ultrasonic microbubbles were used to open blood-brain barriers (BBB) with a reversed and limited behavior feature in the study, which could improve the brain-targeted delivery of anti-tumor drugs. The glioma rat model was prepared. Low-frequency ultrasound was combined with microbubbles to affect the permeability of BBB compared with the permeability of independently administered Evans blue (EB) crossing BBB. Time point and length of ultrasound were investigated whether they affect the permeability of BBB and the damage of brain tissue. The effect of the growth time of glioma on BBB permeability was explored. Only glioma had a very little impact on BBB permeability. However, ultrasonic microbubbles opened the BBB with the features of temporary, limited and reversed behavior and improved EB and magnetic resonance imaging contrast agent penetrating BBB. A length of 30 s ultrasound is appropriate for opening BBB and no damage of brain tissue. Drugs should be injected before ultrasound so that they enter into brain as BBB opening. Ultrasonic microbubbles can open BBB effectively and safely, which improve drugs penetrating BBB under proper time point and length.
Animals ; Blood-Brain Barrier ; Contrast Media ; Drug Delivery Systems ; Glioma ; drug therapy ; Magnetic Resonance Imaging ; Microbubbles ; Permeability ; Rats ; Ultrasonics

OBJECTIVE: To observe the protective effects of Yanggan Lidan Granules (YGLDG) on carbon tetrachloride (CCl(4))-induced liver damage in mice and to find out its mechanism. METHODS: A model of chronic liver damage was established in mice by intraperitoneal injection of CCl(4). After three weeks, those model mice were treated with low-, medium-, high-dose YGLDG, Danning Tablets and bifendate respectively for four weeks. Then the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in liver tissues were detected. RESULTS: YGLDG could significantly reduce the levels of serum ALT and AST in model mice, and the content of MDA was obviously decreased while the content of SOD was increased in liver tissue. CONCLUSION: The therapeutic effect of YGLDG on mice with CCl(4)-induced liver damage is to relieve the seriousness of liver damage, and its mechanism may relate to reducing peroxidation activity in liver tissue.