Objective To observe the expression of TGF?RⅠ,TGF?RⅡ and Smad7 in oxazolone-induced colitis of mice and to investigate the role of the TGF? signal transduction on pathogenesis of colitis. Methods Balb/c mice were pre-sensitized by skin painting with 0.2 ml 3% oxazolone on day 0 and 1 followed by intrarectal administration of 0.15 ml 1% oxazolone on day 7. The mice were sacrificed after 3 days. Colitis was evaluated by macroscopic and microscopic examination. The expressions of TGF?RⅠ, TGF?RⅡ and Smad 7 were examined by immunohistochemical study and Western blot respectively. All the results were compared with the controls. Results Twenty-four hours after intrarectal administration of oxazolone, the mice presented anorexia, less moving, loose stool, hematochezia or occult blood(+) and weight loss. The macroscopic and microscopic scores in two groups were 0.17?0.41, 2.67?1.03 and 2.33?0.52, 8.17?0.75, respectively. In the normal intestine, TGF?RⅠ, TGF?Ⅱ and Smad7 were mainly co-localized on the upper part of the villus. However, their expression was not only throughout the villus including fundus of crypts, but also in the mononuclear cells of the lamina propria and submucosa in the experimental intestine. The amounts of TGF?RⅠ, TGF?Ⅱ, Smad7 and the ratio of TGF?RⅠ/Ⅱ in control and colitis groups were 3.40?1.25, 21.71?6.97, 8.95?2.12, 0.16?0.01 and 6.49?3.18, 4.40?3.34, 17.92?6.80, 2.14?1.61, respectively. Conclusions Decreased TGF?RⅡ and increased Smad7 expressions indicate the abnormality of TGF? signal transduction in oxazolone-induced colitis. These pathologic and immunologic characteristics may resemble human ulcerative colitis.

Objective:To obtain phage-displayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display,so as to lay a foundation for diagnosis and treatment of cancer.Methods:The membrane proteins were extracted from the specimens of esophageal carcinoma,stomach carcinoma,brain cancer,lung cancer,and spinal cord tumor.The recombinant phage-antibody system was used to construct a single-chain Fv fragment(ScFv)cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein.The specific primers of VH and VL were used to amplify the cDNA of VH and VL,respectively,which were then assembled into ScFv gene with a specially constructed linker DNA.The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma(HeLa cells),sepecific phage-ScFvs were selected by phage displaying and panning in vivo.After four rounds,24 phage-ScFvs,which were identified by PCR,were analyzed immunohistochemically.The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced.Results:Tomors-bearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice.It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice.A ScFv library of 1.6?106 was obtained and a tumor vessel specific phage-ScFv named ScFvH1(VH-linker-VL)was selected from the library.Conclusion:A tumor targeting ScFv library has been successfully constructed and a tumor vessel-specifrc antibody has been identified from the library,which provides a new way for the early diagnosis and therapy of cancer.

Objective: To investigate the targeting and anti-tumor ability of the tumor vessel-specific antibody ScFvH1 selected from phage-ScFv library, and to discuss the application of the antibody in clinical diagnosis and therapy of cancer. Methods: The ScFvH1 gene was inserted into pET-28a(+)/EGFP vector containing green fluorescent protein(GFP) gene and pTIG-Trx vector containing thioredoxin gene; the products were then expressed in E.coli and purified by using Ni-NTA. Tumor-bearing mice model was established by subcutanuous injection of cervical cancer cell line HeLa. The mice were injected with purified ScFv-EGFP fusion protein through vena caudalis and the GFP signals were observed by fluorescent microscope to evaluate the targeting ability of the antibody. Meanwhile, the mice model also received intratumoral injection of purified ScFv-EGFP fusion protein to evaluate the anti-tumor effect of the antibody. Results: Soluble ScFvH1 gene and ScFvH1-EGFP protein were successfully expressed in E.coli; a single band was showed in SDS-PAGE after the purification by Ni-NTA. We found that ScFvH1-EGFP fusion protein was enriched to tumor tissues, but there was only weak fluorescent signal when EGFP protein was injected. No EGFP signal was observed in the lung of tumor-bearing mice. Tumor inhibition experiment showed that the tumor growth in the antibody treatment group was similar to that of the PBS control group. Conclusion: The tumor vessel-specific antibody ScFvH1 selected from phage-ScFv library can specifically target tumor vessels, but it has no obvious inhibitory effect on tumor growth. Our findings pave a way for antibody in cancer diagnosis and treatment.

An increasing number of monopartite begomoviruses are being identified that a satellite molecule (DNAbeta) is required to induce typical symptoms in host plants. DNAbeta encodes a single gene (termed betaC1) encoded in the complementary-sense. We have produced transgenic Nicotiana benthamiana and N. tabacum plants expressing the betaC1 gene of a DNAbeta associated with Tomato yellow leaf curl China virus (TYLCCNV), under the control of the Cauliflower mosaic virus 35S promoter. Transgenic plants expressing betaC1 showed severe developmental abnormalities in both species. Microscopic analysis of sections of both transgenic and non-transgenic N. tabacum leaves showed abnormal outgrowths of transgenic N. tabacum to be due to disorganized cell division (hyperplasia) of spongy and palisade parenchyma. Immuno-gold labeling of sections with a polyclonal antibody against the betaC1 protein showed that the betaC1 protein accumulated in the nuclei of cells. The possible biological function of the betaC1 protein was discussed.
Cell Division ; physiology ; Cell Nucleus ; genetics ; metabolism ; ultrastructure ; Cells, Cultured ; DNA, Viral ; genetics ; Geminiviridae ; genetics ; Plant Diseases ; genetics ; virology ; Plant Leaves ; cytology ; genetics ; growth & development ; metabolism ; Plants, Genetically Modified ; growth & development ; metabolism ; Recombinant Proteins ; metabolism ; Tobacco ; cytology ; growth & development ; metabolism ; ultrastructure ; Viral Proteins ; genetics ; metabolism

Objective:To study the feasibility of extraction of impacted mandibular wisdom teeth using turbine drill and new instruments. Methods: 600 patients with impacted mandibular third molars were divides into 2 groups. A group used turbine drill and new instruments to extract the impacted mandibular third molar. B group used the dental chisel to extract the impacted mandibular third molar. The operation time, intraoperative and postoperative complications were recorded to assess the effects of the methods. Results: The operation time of group A and group B was (22.285±12.025 01) min and (16.115±12.078 62) min respectively. The operation time of group A was shorter(P<0.05). The intraoperative and postoperative complication incidence rate was lower(P<0.05). Conclusion: Turbine drill and new instruments method is superior to dental chisel method in the extraction of impacted mandibular wisdom teeth.

Re-evaluation of bioequivalence of generic drugs is one of the key research focus currently. As a means to ensure consistency of the therapeutic effectiveness of drug products, clinical bioequivalence has been widely accepted as a gold standard test. In vitro dissolution testing based on the theory of the BCS is the best alternative to in vivo bioequivalence study. In this article, the conventional dissolution method and flow-through cell method were used to investigate the dissolution profiles of domestic amoxicillin capsules in different dissolution media, and the absorption behavior of the drugs with different release rates (t85% = 15-180 min) in the gastrointestinal tract was predicted by Gastro Plus. The flow-through cell method was thought better to reflect the release characteristics in vivo, and amoxicillin capsules with regard to the release rates up to 45 min (t85% = 45 min) were having a satisfied bioequivalence with the oral solution according to the C(max) and AUC. Although two different dissolution profiles of domestic amoxicillin capsules were found by flow-through cell methods, prediction results revealed that domestic capsules were probably bioequivalent to each other.
Amoxicillin ; pharmacokinetics ; Capsules ; Computer Simulation ; Gastrointestinal Tract ; Humans ; Software ; Solubility ; Therapeutic Equivalency

Powder X-ray diffraction (PXRD) technology combined with cluster analysis method was used to classify 75 batches of crystalline ceftriaxone sodium into subtypes, the crystalline characteristics of each subtype were measured with scanning electron microscope (SEM). By comparing some parameters of these subtypes correlated to crystallization process of ceftriaxone sodium, such as salification rate, water content in different subtypes, as well as by studying different lattice stabilities, different compatibilities with rubber closures during accelerated stability tests, the key point to improve the quality of domestic ceftriaxone sodium was disclosed. The results of this paper indicated that the fine structure of the products could be controlled well by improving the salification and crystallization process. As a result, the subtype II of ceftriaxone sodium with high stability can be produced.
Ceftriaxone ; chemistry ; classification ; Crystallization ; Microscopy, Electron, Scanning ; Powders ; Water ; X-Ray Diffraction

Objective To investigate ultrasonic findings of pure invasive micropapillary carcinoma (PIMPC) of breast. Methods A total of 18 patients with surgically confirmed PIMPC and 40 patients with surgically confirmed invasive ductal carcinoma (IDC) treated between January 2010 and August 2015 in Affiliated Union Hospital of Fujian Medical University, who had undergone preoperative ultrasound examination, were included in the study. To compared with the postoperative pathological examination, the value of ultrasonography in the diagnosis of axillary lymph node metastasis was discussed.Ultrasound findings of PIMPC and pathological results were compared. Results Ultrasound analysis of PIMPC masses identified predominantly hypoechoic lesions and irregular shape 100% (18/18), obscure lesion boundaries 88.9% (16/18), spiculated or angular margins 83.3% (15/18), combined microcalcifications 83.3% (15/18), with posterior acoustic enhancement or normal 88.9% (16/18), dcrab claws changes 77.7% (14/18),witout hyperechoic halo72.2% (13/18) and with 0- Ⅰ grade flow signals 55.6% (10/18). Compared to the IDC, the PIMPC had lower proportions in long speculation, hyperechoic halo, aspect ratio ≥ 0.7, posterior echo attenuation, Ⅱ- Ⅲ grade blood flow signals (P < 0.05), while their lesions in the maximal tumor size,shape, boundary, edge bur, microcalcification has no significance (P>0.05). The rate of lymph node metastasis of PIMPC was 72.2% (13/18), which was significantly higher than that of IDC 45.0% (18/40) (t=3.697,P=0.05). 13 cases were pathologically confirmed lymph nodes metastasis in 18 cases, and among them, the ultrasound indicated abnormal 46.2% (6/13), and showed abnormal cortex and medulla structure 30.8% (4/13), eccentric lymph door 30.8% (4/13), poor blood flow signals 38.5% (5/13). The sensitivity, specificity,positive predictive value, negative predictive value and accuracy of PIMPC lymph node metastasis by preoperative ultrasound were respectively 46.2%, 60.0%, 75.0%, 30.0%, 50.0%. Ultrasound performance of PIMPC has a certain relationship with its special pathological characteristics. Sonography findings were compared with pathological results: Microscopically, PIMPC cell arranged as pseudo-papillary or tubuloalveolar structures floating in empty spaces; PIMPC with 0-Ⅰ grade flow signals were seen that the small amount of new blood vessels of the tumor were mainly found in the pellucid zone around the cell clusters under the microscope, while IDC with Ⅱ - Ⅲ grade blood flow signals were found that more regenerated blood vessels were distributed in the collagen fibers. PIMPC witout hyperechoic halo were noticed that under the microscope, there were no fibrous tissue clusters in the edge of the tumor, meanwhile, IDC with hyperechoic halo were discovered that fibrous tissue was seen at the edge of the mass of the lens. Conclusions Ultrasound performance of PIMPC has a close relationship with its special pathological characteristics. To be familiar with ultrasound characteristic of PIMPC is significant for improving its ultrasound detection rate.

OBJECTIVETo investigate the nephrotoxic effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs.

METHODBeagle dogs were randomly divided into negative control group(blank tablet), methyl cantharidimide tablets group (6.11,12.21, 24.42 mg x kg(-1)), continuously 30 days of oral adminiStration, once a day. The drug and control group were collected and determined fresh urine in 1, 2, 3 and 4 weeks of the administration; Serum urea nitrogen (BUN), creatinine (Crea), total protein (TP) and albumin (ALB) as well as sodium, potassium, chloride electrolyte were determined on 15 and 30 days of the administration; Urine albumin (mAlb), kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin( NGAL), N-acetyl-beta-D-glucosaminidase (NAG), clusterin, beta2-microglobulin (beta2-MG), alpha1-microglobulin (alpha1-MG), alanine aminopeptidase( AAP) and im- munoglobulins IgG were tested on 15 and 30 days of the administration.

RESULTCompared with the control group, urine protein and white blood cells was significantly increased in each dose group. On 15 days of the administration, mAlb were higher in each dose group, KIM-1, NGAL, clusterin, NAG and AAP were significantly higher in high-dose group, while the middle and low dose group had no significant difference, as well as blood SCr and BUN no obvious abnormalities. On 30 days, mAlb, KIM-1, clusterin, NAG, AAP were increased in each dose group, appearing dose-effect relationship, beta2-MG and NGAL levels were significantly increased in high-dose group. Contents above indicators were increased with significant dose and time relationship, and serum BUN, Scr were correlated, suggesting that urine mAlb, KIM-1, clusterin, NAG and AAP indicators that can sensitively respond the changes of proteins and enzymes in urine.

CONCLUSIONMethyl cantharidimide tablets has a renal toxicity, urine mAlb, KIM-1, clusterin, NAG and AAP can be used as the early nephrotoxic biomarkers of methyl cantharidimide tablets.

Animals ; Biomarkers ; urine ; Dogs ; Female ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; Male ; Proteins ; metabolism ; Tablets ; adverse effects ; Urine ; chemistry