Crystal structures of chemical drugs has been being investigated widely. But few attention has been paid to polymorphs-phenomena of active ingredients from Traditional Chinese Medicine(TCM). Taking anhydrous dehydroandrographolide and hydrousprim-O-glucosylcimifugin as example, differences between TCM reference substances (RSs) with different crystal structures were discussed by using microscopy, melting point determination, differential thermal analysis (DTA) and infrared (IR) methods. The results showed that different crystal structures could lead to change of melting points, thermal behaviors and IR spectrum. It's indicated that polymorphs may be considered if different physicochemical properties were obtained when applying TCM RS. Differences of chemical properties of active ingredients from TCM with different crystal structures need further investigation.
Crystallization ; Differential Thermal Analysis ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; Medicine, Chinese Traditional ; Molecular Structure ; Reference Standards ; Spectrophotometry, Infrared ; methods ; standards ; Transition Temperature

Objective To investigate the expression of syndecan-1 (SDC1) in glioma cells and the effects of synde?can-1 knockdown on the proliferation and invasion of A172 cells. Methods The expression of syndecan-1 in glioma cells was analyzed using quantitative Real-time PCR and Western blotting. A172 cells were transfected with lentiviral vector carrying SDC1 shRNA to establish a stable SDC1-silencing cell line. The cell proliferation was analyzed by MTT assay. Trypan blue exclusion assay and flow cytometry, and Transwell assays were performed to measure the migration and invasion abilities, respectively. The mRNA and protein and expression levels of SDC1, Proliferation Cell Nuclear An?tigen (PCNA) and Matrix Metalloproteinase 9 (MMP-9) were detected by using qRT-PCR and Western blotting. Results The expression levels of SDC1 were significantly different in different glioma cell lines. The stable SDC1-silencing cell line was successfully established, in which the mRNA and protein expression levels of SDC1 were significantly decreased (P<0.05). SDC1 knockdown significantly reduced the cell proliferation, migration(58.40±5.24 vs. 255.8±16.09、226.5± 22.84,F=126.4,P<0.05)and invasion(61.67 ± 16.26 vs. 233.70 ± 17.24、244.30 ± 28.15,F=69.87,P<0.05)compared with either control group or blank group. SDC1 knockdown also significantly decreased the mRNA and protein expression levels of PCNA and MMP-9 (P<0.05). Conclusion:SDC1 knockdown suppresses the capacities of proliferation, invasion and migration of glioma A172 cell, implying that SDC1 may serve as a novel target in the biotherapy of glioma.

Objective To investigate the effect of different gene expression levels of Syntenin on invasion and mi?gration of glioma cells and the underlying molecular mechanisms. Methods Lentiviral RNA interference was used to knockdown the expression of syntenin in U-87 cells. Real-time quantitative PCR was used to detect mRNA expression levels of syntenin . Transwell assay and adhesion assay were used to examine the invasion, migration and adhesion, re?spectively. Western-blot was used to detect the protein expression levels of Syntenin, AKT, p-AKT, and MMP-9. Re?sults The mRNA expression level of Syntenin was greatly reduced in interference group compared with empty vector group (P<0.01). The ability of invasion and migration was much lower in interference group than in empty vector group (P<0.01). However, there were no significant differences in invasion and migration between empty vector group and con?trol group. The adhesion ability of glioma U-87 cells was much higher in interference group than in empty vector group (P<0.05). However, when U-87 cells were seeded on 96-wells coated with HUVEC, the adhesion ability was much lower in interference group than in empty vector group(P<0.05). The protein expression levels of Syntenin, p-AKT, and MMP-9 in interference group were markedly decreased compared with empty vector group(P<0.05). There was no signif?icant difference in expression of AKT protein between interference group and empty vector group (P>0.05). Conclusion Syntenin may enhance the invasion and migration ability of glioma though up-regulation of p-AKT, which in turn pro?motes MMP-9 expression in a corresponding signal transduction pathway.

Objective The aim of the study was to investigate the expression of P-glycoprotein (P-gp)and nuclear factor κB (NF-κB) in the cerebral cortex of rat with chronic fluorosis,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of fluoride.Methods Sixty SD rats were randomly divided into three groups.The rats in each group were given drinking water containing different levels of fluoride:control group less than 0.5 mg/L,small amount of fluoride exposure group 10.0 mg/L and large amount of fluoride exposure group 50.0 mg/L.The animals were examined at the sixth month after initiating the experiment.Protein levels of P-gp and NF-κB in brain tissues were detected by immunocytochemistry and Western blotting,and the P-gp protein and mRNA level by quantitative real time PCR method.Results As compared to the control group(28.21 ±6.13),the numbers of positive staining cells by P-gp antibody in the cortex of rat brains were significantly increased in both the small and the large amount of fluoride exposure groups[(48.46 ± 8.00),(53.72 ± 9.15),respectively,all P ＜ 0.05] ; the protein levels in the control group(100.00 ± 3.86)％ detected by Western blotting were significantly increased in the cortex of rat brains treated with fluoride in both the small and the large amount of fluoride exposure groups[(189.47 ± 3.14)％,(191.36 ± 11.09)％,respectively,all P ＜ 0.05].The significantly increased expression of NF-κB at the protein level was observed in the cortex of rat brains of the small and the large amount of fluoride exposure groups[(365.97 ± 6.04)％ and (417.15 ± 10.89)％,respectively] as compared with the control group[(100.00 ± 10.07)％,all P ＜ 0.05].The mRMA level of P-gp in the cortex of rat brains of the small and the large amount of fluoride exposure groups(2396 ± 427,3479 ± 371,respectively) were higher than that of the control group(260 ± 106,all P ＜ 0.05).Conclusion The increased expressions of P-gp and NF-κB in the cortex of rat brains are induced by chronic fluorosis,which might be connected with the mechanism of brain damages.

Objective To explore the impacts of over-expression of microRNA-7 (miRNA-7) on the sensitivity of cis-platin in esophageal carcinoma cell line TE-1, and the possible mechanism thereof. Methods Lipofectmin 2000 method was used to transient transfect with miRNA-7 mimic into esophageal cancer cell line TE-1, which was taken as transfection group, mimic negative control was taken as transfection conrtol group. The expressions of miRNA-7 and epidermal growth factor receptor (EGFR) mRNA were detected by RT-PCR in the above two groups and normal control group. The total EGFR and EGFR in cytoplasmic and nucleus were detected with Western blot assay in transfection group and transfection control group. CCK-8 was used to detect IC50 of cisplatin in transfection group and transfection control group. The expression of EGFR was observed with immunofluorescence confocal microscope in two groups. Results The miRNA-7 expression was signifi-cantly increased in transfection group than that of transfection conrtol group and control group. The expression of EGFR mRNA was significantly reduced in transfection group (P<0.001). The total EGFR was significantly decreased in transfec-tion group than that of transfection conrtol group. The level of nuclear EGFR was significantly increased ( P<0.01),and cyto-plasm EGFR expression was significantly decreased in transfection group than that of transfection control group ( P<0.05). CCK-8 results showed that after the over expression of miRNA-7 in TE-1, the IC50 of cisplatin (48 h) increased in transfec-tion group than that of control group (P<0.01). Immunofluorescence results showed that EGFG in nuclear was higher in transfection group than that of transfection control group but its expressions reduced in cell membrane and cytoplasm. Con-clusion The over-expressed miRNA-7 in esophageal cancer cells TE-1 can reduce cisplatin sensitivity by the increased EGFR in nuclear translocation.

OBJECTIVETo investigate diagnosis and therapeutic effects of knee joint synovial chondromatosis with arthroscopic.

METHODSFrom March 1995 to July 2011, 28 patients with knee joint synovial chondromatosis were treated. Among them, 18 males and 10 females ranging age from 25 to 81 (mean 55.2) years,the course of disease ranged from 0.5 to 15 (mean 5.6) years. Clinical manifestation mainly included pain, swell and functional limitation of knee joint. Knee open surgery (17 cases) and laparoscopic surgery (10 cases) were respectively used. Clinical symptom,image data,pathological manifestation and effects under arthroscopy were observed, Lysholm scoring was used to evaluate effects.

RESULTSAll patients were followed up except one lost, the duration ranging from 6 to 24 months. Lysholm score in knee open surgery was increased from (41.89 +/- 6.81) preoperatively to (67.73 +/- 7.62) postoperatively;while in laparoscopic surgery it was increased from (40.78 +/- 7.54) preoperatively to (77.46 +/- 8.43) postoperatively.

CONCLUSIONArthroscopic surgery, which has no risk of rupture of incision, nonunion, earlier to exercise, is a good method to diagnosis and treat knee joint synovial chondromatosis.

Adult ; Aged ; Aged, 80 and over ; Chondromatosis, Synovial ; diagnosis ; surgery ; Female ; Follow-Up Studies ; Humans ; Knee Joint ; surgery ; Laparoscopy ; Male ; Middle Aged ; Treatment Outcome

Objective To study the protective effect and mechanism of Inonotus obliquus polysaccharide on liver injury induced by isoniazid( INH) combined with rifampicin( RFP) in mice. Methods Forty-eight mice were randomly divided into six groups ( control, model, huganpian, Inonotus obliquus polysaccharide at low, middle, and high dose ) . The mice were administered orally with isoniazid and rifampicin simultaneously except the control. After 2 h,the control and model groups were administered with normal saline,and others were treated with huganpian and Inonotus obliquus polysaccharide,respectively,once a day for 2 weeks. The liver index and serum levels of ALT and AST,malondialdehyde(MDA),superoxide dismutase(SOD) and glutathione peroxidase( GPx) were measured. Pathology examination of liver tissue was performed. Results The activities of ALT and AST, liver index, content of MDA in liver homogenate of mice treated with Inonotus obliquus polysaccharide and huganpian decreased obviously, while the liver SOD activity increased. Histopathological exzamination showed that Inonotus obliquus polysaccharide alleviated the degeneration and necrosis of hepatic cells. Conclusion Inonotus obliquus polysaccharide shows protective effects on mice with hepatic toxic injury induced by isoniazid and rifampicin,which maybe due to its antioxidation effect.

ObjectiveTo investigate the transcriptional changes of nitochondria fission and fusion gene loci and reactive oxygen species (ROS) level in cortical neurons of rats with chronic fluorosis,and to reveal their roles in mitochondria damage due to chronic fluorosis.MethodsSD rats were fed with different doses of fluoride through drinking water[＜ 0.5(control),10,50 mg/L,respectively] for 3 and 6 months.The level of ROS and mRNA contents of mitochondria fission gene loci Drp1/Fis1 and fusion gene locus Mfn1 in the cortical neurons of rat brains were detected with ROS fluorescent probe and real-time PCR,respectively.ResultsAs compared with control group [10.43 ± 5.98,(3.4 ± 0.6) × 103,(8.8 ± 1.4) × 10,(1.2 ± 0.2) × 102] at the experiment period of 3 months,the level of ROS and mRNA contents of mitochondria fusion gene locus Mfn1 and fission gene loci Drp1/Fis1 in the cortical neurons were obviously increased in the rats fed with 50 mg/L fluoride through drinking water[25.48 ± 6.09,(1.0 ± 0.2) × 1011,(3.0 ± 1.6) × 103,(8.9 ± 3.6) × 102,all P ＜ 0.05],whereas no significant changes were found in the rats fed with 10 mg/L fluoride[11.67 ± 3.49,(3.1 ± 0.3) × 104,(6.7 ± 2.7) × 10,(5.0 ± 0.9) × 10,all P ＞0.05].Furthermore,at 6 months of the experiment the increases in ROS level(63.02 ± 8.15,65.60 ± 7.40) and mRNA contents of mitochondria fission gene loci Drp1/Fis1 [(2.0 ± 0.8) × 106,(4.0 ± 0.6) × 105,(3.8 ± 1.3) × 103,(1.3 ± 0.2) × 103] and the decrease in mitochondrial fusion gene locus Mfn1[(3.0 ± 0.4) × 106、(4.0 ± 0.9) × 104]were observed in the cortical neurons of the rats fed with 10 mg/L and 50 mg/L fluoride as compared with the control group[25.26 ± 6.41,(3.0 ± 0.8) × 109,(5.1 ± 0.8) × 103,(2.8 ± 0.7) × 102,all P ＜ 0.05].Conclusions Excessive intake of fluorine leads to elevated ROS levels,and decreased transcription of mitochondrial fusion gene loci Mfn1,which is positively correlated with the time and dose-exposed to fluoride.The changes of mitochondrial fission and fusion gene loci in the cortical neurons may be related to high level of oxidative stress induced by chronic fluorosis.

Objective To investigate the deletion pattern of 4.8 kb mitochondrial DNA(mito-DNA) in liver,kidney,and brain of rats with chronic fluorosis and to explore the significance of mitochondria in the pathogenesis of chronic fluorosis.Methods Sixty SD rats were randomly divided into 3 groups according to body mass (20 in each group):control,low-fluoride and high-fluoride groups,and they were fed with different concentrations of fluoride in drinking water (0,10,50 mg/L,respectively) for 6 months.Mito-DNA in liver,kidney and brain was detected by real-time PCR.Results The amounts of 4.8 kb mito-DNA in liver(2.1 × 10-3,1.6 × 10-3),kidney (1.7 × 10-3,1.4 × 10-4) and brain cortex (1.5 × 10-5,1.3 × 10-5) in low-and high-fluoride groups were significantly reduced,as compared with that of control group (2.9 × 10-3,2.0 × 10-3,1.1 × 10-4,all P ＜ 0.05).The amount of 4.8 kb mito-DNA in kidney in high-fluoride group was lower than that in low-fluoride group (P ＜ 0.05).Conclusions Excessive fluoride intake can result in missing of 4.8 kb mito-DNA in liver,kidney and brain cortex.The abnormal of mito-DNA might be related to the dysfunction of mitochondrial respiratory chain.

OBJECTIVETo evaluate the impact of hyphema secondary to high intraocular pressure on corneal pathology in rabbits.

METHODSThirty adult New Zealand rabbit were randomized into 3 equal groups, and in each rabbit, one eye served as the experimental eye with the other as the control eye. In the experimental eye, autoblood was injected into the anterior chamber to induce high intraocular pressure maintained for 3, 5, or 8 days. Only saline was injected into the control eye. After the injections, the cornea was observed with slit-lamp microscopy, and at 3, 5, or 8 days, the experimental and control eyes were taken from the 3 groups for microscopic examination of the corneas to detect the occurrence of cornea bloodstain with prolonged high intraocular pressure. Corneal edema, elastic fibers changes, growth of new blood vessels, changes of eosinophils, fibroblasts, lymphocytes and plasma cells, as well as the pathological changes of the corneal layers were observed and compared between the experimental and control eyes.

RESULTSMaintenance of high intraocular pressure for 8 days resulted in the most severe corneal edema and thickening, and histopathologically, the corneal stroma showed widened space between the elastic fibers and obvious fiber distortion. Neovascularization was seen in the marginal cornea where eosinophil infiltration occurred with a small number of lymphocytes, plasma cells and fiber cells. All the three groups showed more obvious edema in the posterior than in the anterior cornea.

CONCLUSIONProlonged hyphema with ocular hypertension results in aggravation of corneal edema, and corneal blood staining does not occur until 8 days of high intraocular pressure but corneal elastic fiber disruption can be seen, suggesting the impending irreversible pathological changes of cornea.

Animals ; Cornea ; pathology ; Edema ; pathology ; Female ; Hyphema ; complications ; pathology ; Male ; Ocular Hypertension ; complications ; pathology ; Rabbits ; Random Allocation