Objective To establish antibody titre quantitation method by ELISwithoustandard substance .MethodThe tesgroupof non-specifi,negative control ,specifiand total antibodiewere sefodetecting serantibodieby ELIS.Aftelineafitting of the time-absorbance datof early developing ,the fitted slopewere used athe velocity of absorbance changing , which were denoted by ν0 ,νC,νS,νT.Based on thathe ν-value and the concentration ? of determinand were linearelationship while the substrate waexcessive ,the function with parameteof ν-valueforeflecting the multiple proportionof antibodiecon-centrationbetween specifiand negative control groupcould be deduced .The assessmenof specifiIgG antibodiein serof KM mice immunized with fish collagen waused aan instance .ResultThe function C/C= (ν-ν0 )/(νC-ν0 ) could calculate the mul-tiple proportion(titre) of antibodieconcentrationbetween specifiand negative control group.Conclusion The above method of antibody titre quantitation isuitable fosemi-quantitative analysiwithoustandard substance .

Objective To observe the dynamic expressions of kielin/chordin-like protein (kcp) in mouse model of hepatic fibrosis and the effects of bone morphogenetic protein-7 (BMP-7)intervention on the expressions,and to explore a new target for treatment of fibrosis.Methods A total of 50 healthy male ICR mice were divided into three groups:control group(n=10) ; model group (n=30) and BMP-7 treatment group (n=10).The model group was further divided into three subgroups according to different time points:subgroups of 4,8 and 12 weeks with 10 mice in each subgroup.The mouse model of hepatic fibrosis was established by hypodermic injection of carbon tetrachloride(CCl4 ).The mice in BMP-7 treatment group began to receive human recombmant BMP- 7via intraperitoneal injection after 8 weeks of the first administration of CC14 and lasted for 4 weeks.The serum levels of alanine aminotransferase (ALT),aspartate transaminase (AST) and albumin (Alb) were detected.The pathological changes of liver were observed under optical microscope after HE and Masson staining.The dynamic expressions of kcp mRNA and protein of each group were detected by reverse transcription-polymerase chain reaction,immunohistochemistry and Western blot.The comparison of means among groups was done by univariate ANOVA.Results In model group,ALT and AST levels increased,while Alb level gradually decreased,and peaked at week 12.BMP-7treatment could reduce the changes,and there were significant differences among groups (F=23.501,34.600 and 16.244,respectively; all P＜0.05).In normal control group,the expressions of kcp mRNA and protein were low,while those in model group were gradually increased.BMP-7 treatment could achieve remission and the changes were all significantly different among groups (F=30.362 and 10.727,respectively; P＜0.01 or 0.05).The expression of kcp mRNA was positively correlated with levels of transforming growth factor (TGF)-β1 mRNA and BMP-7 mRNA (r=0.760 and 0.769,respectively; both P＜0.05).Conclusions BMP 7 can improve the hepatic fibrosis in mice.kcp may play an important role in the occurrence and development of liver fibrosis which is a potential therapeutic target for hepatic fibrosis.

Researching on a large relevant literature of spinal tuberculosis, this paper discussed characteristics of spinal tuberculosis and its early diagnosis method. If spinal tuberculosis can be found and treated earlier, prognosis will be better. The early diagnosis of spinal tuberculosis has become hot spot at present. There are many different diagnostic methods including clinical manifestation, laboratory examination, imaging examination, etc. The newest standpoint showed that immunization has closely correlation with tuberculosis and it will become the focus and developmental direction of future research in spinal tuberculosis. In addition, we should know the differential diagnosis of spinal tuberculosis to avoid the misdiagnosis.
Diagnosis, Differential ; Early Diagnosis ; Humans ; Tuberculosis, Spinal ; diagnosis

AIM To observe the protective effect of Ganduqing Granule(GDQ) on acute hepatic injury. METHODS Sixty male Wistar rats were divided into 6 groups, namely normal control group, model group, Ganduqing large (GDQ Ⅰ), middle(GDQ Ⅱ), small dose(GDQ Ⅲ) groups and Yiganning (YGN) positive group. Acute hepatic injury induced by thioacetamide. The changes of blood serum ALT, AST activity and plasma endotoxin (ET) level and TNF-?, IL-6 levels were measured and livers were taken for pathology examination. RESULTS Plasma ET level and blood serum ALT, AST levels and TNF-?, IL-6 levels were significantly lower than model group ( P

OBJECTIVETo investigate the protective effect and mechanism of curcumin derivatives B06 on myocardium from type 2 diabetic rats.

METHODSThirty-five male SD rats were randomly divided into 5 groups, normal control group (NC group), high fat group (HF group), high fat treatment group (FT group), diabetes mellitus group (DM group) and diabetes treatment group (DT group) (n = 7). The late four groups were fed with high fat food, after four weeks of high fat feeding, the rats from DM group and DT group were injected with low dosage of streptozocin intraperitoneally to induce diabetes mellitus, FT group and DT group were gavaged with curcumin derivatives B06 at the dosage of 0.2 mg/kg x d. The blood glucose and lipid were detected biochemically, blood insulin was assayed by ELISA and the insulin resistance index was calculated, the morphology of myocardium was observed by light and transmission electron microscopy, the protein expression of AMP-activated protein kinase alpha (AMPKalpha) and phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) in myocardium were tested by Western blot.

RESULTSThe level of blood glucose, lipid, insulin and the insulin resistance index were increased in HF group and DM group, but they were decreased after the treatment with B06. The expression of AMPKalpha and p-AMPKalpha were decreased, but they became increased after the treatment of B06. There were increased collagen fibers in interstitium and expansion of mitochondria in cytoplasm of myocardium from DM group, but they were ameliorated in B06 treatment group.

CONCLUSIONIt is suggested that B06 may relieve the damage of myocardium from type 2 diabetic rats and the increased expression of AMPKalpha and p-AMPKalpha may be involved in it.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Blood Glucose ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; Insulin Resistance ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

This study was aimed to investigate the effects of glycyrrhetinic acid (GA) on proliferation, apoptosis and survivin mRNA expression in human myeloma cell line U266 in vitro. Cell proliferation was assayed by MTT method. Both cell apoptosis and cell distribution in cell cycle were analyzed by using flow cytometry. Scanning electron microscopy was used to observe the morphological changes in U266 cells induced by GA. Expression of survivin mRNA was detected by quantitative real-time reverse transcription-polymerase chain reaction. The results showed that GA inhibited proliferation of U266 cells in a time- and dose-dependent manners in vitro. GA presented apoptosis induction potency to U266 cells, obvious changes in morphology of U266 cells was observed under scanning electron microscope. The cells were arrested at the G(0)/G(1) phase, showing the accumulation in G(0)/G(1) phase, reduction of cells in G(2)/M phase and S phase. GA could down-regulate the expression of survivin gene in U266 cells in a dose-dependent manner. It is concluded that GA can inhibit proliferation of U266 cells in a time- and dose-dependent manners and induce apoptosis of this cell line in vitro through arresting G(0)/G(1) phase and down-regulating expression of survivin.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Glycyrrhetinic Acid ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology

OBJECTIVETo investigate the effect of Shuyusan decoction on neuropeptide Y (NPY) and serotonin (5-HT) expression in the hippocampus and plasma of rats with chronic mild unpredictable stressors depression.

METHODSFifty Wistar rats were randomly divided into 5 groups, namely the normal control group, model group, fluoxetine group, and high- and low-dose Shuyusan groups. Except for those in the normal control group, all the rats were subjected to chronic mild unpredicted stress for 21 consecutive days with corresponding treatments. Open-field test was used to assess the behavioral changes of the rats. The content of NPY in the hippocampus and plasma was detected by competitive enzyme-linked immunosorbent assay, and immunocytochemistry was used to determine the expression of 5-HT in the hippocampus.

RESULTSNPY levels in the hippocampus and plasma was significantly decreased in the model group as compared with that in the normal control group (P<0.05). Treatments with fluoxetine and high-dose Shuyusan both significantly increased NPY levels in the hippocampus and plasma in the depressive rats (P<0.05), resulting also in significantly increased 5-HT-immunoreactive neurons in the cerebral cortex and hippocampus and the average optical density (P<0.05).

CONCLUSIONShuyusan decoction can effectively increase plasma and hippocampus NPY levels and the number of 5-HT-positive neurons in the cerebral cortex and happocampus of rats with chronic mild unpredictable stress-induced depression.

Animals ; Depression ; etiology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hippocampus ; metabolism ; Male ; Neurons ; metabolism ; Neuropeptide Y ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Serotonin ; metabolism ; Stress, Physiological

OBJECTIVETo express and purify Schistosoma japonicum ribosomal protein S4(SjRPS4) in Escherichia coli, and assess its value in immunodiagnosis of Schistosomiasis japonica.

METHODSGene fragment of SjRPS4 was amplified by screening the cercaria cDNA library of Schistosoma japonicum. The target gene was cloned into the expressive vector pQE30 and transformed into E. coli M15. The recombinant protein expression was induced by isopropylthio-β-D-galactoside (IPTG). This fusion protein was purified by Ni(2+)-NTA chromatography and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and ELISA.

RESULTSThe plasmid pQE30/SjRPS4 was constructed successfully and expressed a SjRPS4 fusion protein in E. coli as showing a single special band on SDS-PAGE gel at Mr 30 × 10(3) position. It reached a purity of above 90% after purification. The Western blot result confirmed that the recombinant protein could specifically react with the serum samples from patients of schistosomiasis. Detecting the serum of Schistosomiasis japonica patients by ELISA, the sensitivity and specificity of the ELISA method were 90.91% (70/77) and 92.59% (25/27), the positive rate of recombinant protein expression was 67.30% (70/104). There was no cross-reaction with paragonimiasis patients' serum.

CONCLUSIONProtein SjRPS4 was successfully cloned and expressed, and it was confirmed that SjRPS4 antibodies were valuable in the diagnosis of Schistosomiasis japonica.

Amino Acid Sequence ; Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Library ; Humans ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; genetics ; Ribosomal Proteins ; genetics ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; diagnosis ; genetics ; Sensitivity and Specificity

OBJECTIVEThis study explored the dynamic expression of the E3 ubiquitin-protein ligase gene, Arkadia, in response to carbon tetrachloride (CCl4)-induced liver fibrosis in a mouse model and investigated the differential expression that occurs following treatment with the anti-fibrotic bone morphogenetic protein-7 (BMP-7).

METHODSThirty healthy male imprinting control region (ICR) mice were randomly assigned to three groups: normal (control; n = 6), CCl4-induced model group (model; n = 18), and CCl4-induced model with BMP-7 treatment group (treatment; n = 6). The model group was further divided into three subgroups (n = 6 each) for analysis at 4, 8 and 12 weeks after fibrosis induction. Liver fibrosis was induced by hypodermic injections of 60% CCl4 /peanut oil (5 mL/kg) to the hind legs of mice two-times per week in alternating legs for a period of 12 weeks. At week 9, the treatment group of CCl4-induced mice were given an intraperitoneal injection of BMP-7 (300 pg/g) simultaneously with that day's hypodermic injection of 60% CCl4 /peanut oil, and then every other day for a period of four weeks. The pathological changes in liver tissues were observed after staining with hematoxylin-eosin (HE) and Masson's trichrome. Messenger RNA (mRNA) and protein expression of Arkadia in liver were evaluated using reverse transcription-polymerase chain reaction and immunohistochemistry and Western blotting, respectively.

RESULTSMouse models of liver fibrosis were successfully established by CCl4 exposure. Arkadia, Smad7 and TGF-beta1 mRNA levels were up-regulated in the model group in a time-dependent manner (vs. control group), and BMP-7 treatment led to significant down-regulation of the CCl4-induced expression of the three genes (vs. control group: F = 812.80, 451.46, and 998.96, respectively; P less than 0.01). At week 12, the mRNA levels of Arkadia, Smad7, and TGF-b1 were significantly lower in the BMP-7 treatment group than in the model group (t = 12.108, 18.737, and 16.364, respectively; P less than 0.01). Arkadia, Smad7, and TGF-b1 protein staining was weak in the portal area of control liver tissue. In contrast, the model group showed significantly stronger staining for all three proteins in the portal area and in the cytoplasm of liver cells. The staining of Arkadia, Smad7, and TGF-b1 proteins was significantly lower in the treatment group (vs. control group: F = 8.399, 609.690, and 900.561, respectively; P < 0.01). At week 12, the protein levels of Arkadia, Smad7, and TGF-b1 were significantly lower in the treatment group than in the model group (t = 23.438, 11.667, and 42.889, respectively; P < 0.01).

CONCLUSIONArkadia expression gradually increased along with the development of liver fibrosis but was suppressed by treatment with the anti-fibrotic factor, BMP-7.

Animals ; Bone Morphogenetic Protein 7 ; pharmacology ; Liver ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; prevention & control ; Male ; Mice ; Mice, Inbred ICR ; Ubiquitin-Protein Ligases ; metabolism ; Up-Regulation

· AIM:To compare the clinical curative effect of triamcinolone acetonide and Ranibizumab on diffuse diabetic macular edema (DME).· METHODS:We collected 84 cases of patients with diffuse DME treated in our hospital from February 2016 to May 2017.According to 1:1 matching method,they were divided into Ⅰ,Ⅱ groups.They were all treated with laser photocoagulation.Preoperative auxiliary application of triamcinolone acetonide was given to Group Ⅰ,while Group Ⅱ received preoperative application of ranibizumab.After treatment,the efficacy of the two groups were analyzed and compared.· RESULTS:The total clinical efficiency of Group Ⅱ at 3mo after treatment was 93％,higher than that of Group Ⅰ (77％;x2 =4.981,P =0.025).Compared with before treatment,BCVA and CMT of the two groups at each time after treatment were significantly improved (P＜ 0.05).BCVA of Group Ⅱ at 1 and 3mo after treatment was better than that of Group Ⅰ (P＜0.05);CMT of Group Ⅱ at 1,3 and 6mo after treatment improved more than that of Group Ⅰ,with significant difference (P ＜ 0.05);occurrence rate of adverse reactions Group Ⅰ and Group Ⅱ were 17％ and 13％ with no significant statistical difference (X2 =0.243,P=0.621).There were no serious adverse reactions such as retinal detachment,endophthalmitis or cataract in the two groups.· CONCLUSION:Compared with triamcinolone acetonide,the effect of ranibizumab on diffuse diabetic macular edema is better,and has high clinical value.