Objective To explore the curative effect of endourological treatment of pediatric urethral hemangioma with holmium laser.Methods Two children with urethral hemangioma were enrolled in this study. One urethral hemangioma in the bulbous urethra,another in the posterior urethra. Modalities of diagnosis before operation included B ultrasound,intravenous urogram(IVU) and endoscopy. Two children with urethral hemangioma underwent holmium laser therapy.Results After endourological trearment with holmium laser, two children had been cured.Six months to 4 years follow-up did not find recurrent urethral hemangioma.Conclusions The diagnosis of pediatric urethral hemangioma mainly rely on endoscopy. Endourological treatment with holmium laser is a minimally invasive,safe and effective methods for pediatric urethral hemangiomas.

In order to identify the chemical constituents of Yushu tablets comprehensively, we studied the chemical constituents of CHCl3 extract from Yushu tablets by the ultra performance liquid chromatography-electrospray ionization-ion trap-time of flight mass spectrometry (UPLC-ESI-IT-TOF/MS). It showed that there were more than 100 compounds separated, and forty-nine peaks among these were identified on the basis of high resolution mass spectrometry data and literature data reported. Determination of twelve peaks was further confirmed by standard substances. These components assigned to the different plant sources mainly included phenylpropanoids, triterpenoids, quinones and m-trihydroxybenzene compounds. By analyzing the chemical components of CHCl3 extract from compound Chinese medicine Yushu tablets comprehensively, this study provided the foundation for studying chemical components, pharmacodynamic substance and quality control of Yushu tablets.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drugs, Chinese Herbal ; analysis ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Tandem Mass Spectrometry

Background Glaucoma is one of the leading causes of blindness worldwide;while the different types of glaucoma is vary from different region. Objective Present study was to survey the prevalence and types of glaucoma among peasants of Uigur adults in Kuche county and offer the basis for the prevent and treatment of glaucoma in Uigur nationanlity. Methods 4191 Uigur peasants aged 40 years or above were collected in Kuche county for the survey of prevalence and types of glaucoma by randomized cluster sampling in March and April of 2009. The subjects were grouped into 40-,50-,60- and ≥70 years groups according to the distribution of age. The disease history of glaucoma, regular eye examination, funds examination and measurement of the anterior chamber depth, gonioscope were performed in the all subjects. Darkroom prone test and mydriasis test were carried out in suspicious glaucomous patients. The depth of periphery anterior chamber was assessed based on van Herick' s criteria, and the width of chamber angle was graded based on Scheie' s method. The standardized training was performed. This survey approved by Xinjiang Medical Ethics Committee, all subjects signed the informed consent before the examination. Results 4191 of 4873 subjects finished all the examinations with the response rate 86%. All the subjects showed a good compliance. The prevalence of glaucoma was 3. 79% , and the prevalence of primary angle-closed glaucoma(PACG) .primary angle-open glaucoma (PAOG) and secondary glaucoma was 2.22% ,0.26% and 1. 31% respectively, showing a significant difference among the different types of glaucoma( P<0. 05). The prevalence of glaucoma was elevated with aging (χ2 - 116. 69 ,P<0. 05) and presented a high rate in male subjects compared with female ones(χ2 = 7. 34, P<0. 05 ). Bilateral blindness was found in 19.75% glaucoma peasants, in which 25.3% glaucoma peasants received anti-glaucoma surgery. The distribution of visual acuity of patients was of significant difference among different age groups(χ2 = 37. 69 ,P<0. 05 ) . Conclusions The prevalence of the glaucoma among Uigur peasants in Kuche county was higher than most area no matter inland or overseas. PACG still is the common type in those people.

Objective By comparing with the SD rats mixed glial cells, C57 mice mixed glial cells and Kunming mice mixed glial cells and exploring the expression of inflammatory factor of three mixed glial cells induced by lipopolysaccharide (LPS), the study aimed to explore the application value of three kinds of mice and find the ideal model of inflammation. Methods We used LPS as inducers, and NO, IL － 1β, COX－2and iNOS as anti－inflammatory antioxidant index. After cutting the head and taking the brain, we cultured the mixed glials. Then we used Greiss assay to detect the expression of NO and used western blot to detect the expression of protein of IL－1β, COX－2, and iNOS protein.Finally we compared the mixed glial cells from SD rats, C57 mice mixed glial cells and Kunming mouse mixed glial cells and selected the best inflammation model from three mixed glial cells. Results The results showed that the morphological changes of the mixed glial cells in SD rats were treated with N2－ free medium. Compared with the control group, the quantity of NO of LPS group of three mixed glial cells increased significantly (P＜0.01) . The LPS group of SD rats released the highest concentration of NO. Western blot was used to detect the expression of IL－1β, Cox－2 and iNOS in three kinds of rodents. Compared with the blank control group, the expression of COX－2 protein, iNOS and IL－1β in the LPS group of the three mice increased significantly. The results showed that LPS could successfully stimulate the release of inflammatory cytokines in three kinds of mice,among which the SD rats were more sensitive and it could be used in the study of AD inflammation model. Conclusion The results showed that LPS could induce the release of NO and the expression of IL－1β, iNOS and COX－2 in C57BL/6 mice,Kunming mice and SD rats to induce inflammatory response. Thus,LPS can induce the formation of inflammatory oxidation models of the original mixed glial cells of the three mice. Moreover, the SD rats were more sensitive.

OBJECTIVETo observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α) in IL-1β induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages.

METHODSNaringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1β-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1β-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1β and TNF-α in IL-1β-induced CDs was detected by RT-PCR.

RESULTSThe appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1β on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α in IL-1β induced CDs (P < 0.05), which was approximate to the level of the normal group.

CONCLUSIONSNaringin could not only promote the proliferation of CDs, but also protect IL-1β-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1β and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

Animals ; Blotting, Western ; Cartilage, Articular ; Caveolins ; Chondrocytes ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; pharmacology ; Interleukin-1beta ; metabolism ; Polypodiaceae ; Rabbits ; Rhizome ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.

METHODSNP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.

RESULTSThree recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (1:2000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1:640 to 1:1280.

CONCLUSIONThe recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.

Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Influenza A virus ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology

In this study, the recombinant adenovirus (Ad) vector containing dual reporter gene [i.e. human transferrin receptor gene (TFRC) and firefly luciferase reporter gene] was constructed to provide a novel experimental tool for magnetic resonance (MR) and bioluminescence dual-modality molecular imaging. The cDNA of TFRC was amplified by polymerase chain reaction (PCR) and cloned into the multiple cloning site of pShuttle-CMV-CMV-Luciferase vector. After identification by Sfi I digestion and sequencing, pShuttle-TFRC-Luciferase vector and the adenoviral backbone vector (pAdeno) were subjected to homologous recombination. The correct recombinant plasmid was then transfected into 293 packaging cells to produce adenoviral particles and confirmed by PCR. After infection of human colorectal cancer LOVO cells with Ad-TFRC-Luciferase, the expressions of transferrin receptor (TfR) and luciferase protein were detected respectively by Western blotting and bioluminescence imaging in vitro. The results showed that TFRC gene was successfully inserted into the adenoviral shuttle vector carrying luciferase gene. DNA sequence analysis indicated that the TFRC gene sequence in the shuttle plasmid was exactly the same as that reported in GenBank. The recombinant plasmid was identified correct by restriction digestion. Ad-TFRC-Luciferase recombinant adenovirus was constructed successfully, and the virus titer was 1.6×10(10) pfu/mL. Forty-eight h after dual reporter gene transfection, the expressions of TfR and luciferase protein were increased significantly (P<0.01). It was concluded that the recombinant adenovirus vector with dual reporter gene was successfully established, which may be used for in vivo tracing target cells in multimodality imaging.
Adenoviridae ; genetics ; Genes, Reporter ; genetics ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Molecular Imaging ; methods

OBJECTIVETo evaluate the effect of Zhuanggu Jianxi Decoction (, ZGJXD) on interleukin-1 β (IL-1 β)-induced degeneration of chondrocytes (CDs) as well as the activation of caveolin-p38 mitogen-activated protein kinase (MAPK) signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis.

METHODSSerum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum (ZGJXD-S) was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL-1 β stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type II collagen immunocytochemistry. After IL-1 β-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 (a specific inhibitor of p38 MAPK) for 48 h, caveolin-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL-1 β, tumor necrosis factor α (TNF-α), matrix metalloproteinase 3 (MMP-3) and MMP-13 were examined by real-time polymerase chain reaction.

RESULTSIL-1 β stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 β, TNF-α, MMP-3 and MMP-13. However, the IL-1 β-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs.

CONCLUSIONZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.

Animals ; Base Sequence ; Blotting, Western ; Caveolins ; metabolism ; Chondrocytes ; drug effects ; enzymology ; metabolism ; DNA Primers ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Profiling ; Interleukin-1beta ; physiology ; MAP Kinase Signaling System ; Male ; RNA, Messenger ; genetics ; Rabbits ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

In this study, chemistry, biology and pharmacology were combinated to screen pseudoallergenic substances of Shuang-huanglian injection (SHLI) so that to establish a scientific and systematic approach to screen pseudoallergenic substances of traditional Chinese medicine injections. The mouse pseudoallergic reaction models were used to screen the pseudoallergic reaction of SHLI's intermediate extract and the intermediate extract's component or ingredient. Among the three intermediates of Shuanghuanglian injection (extract of Scutellaria baicalensis, extract of Lonicera japonica, extract of Forsythia suspensa) , pseudoallergic action of Forsythia suspensa was the strongest, Forsythia suspesnsa's pseudoallergic reaction mainly associated with the composition with largerchemical polarity. Further it was found that forsythiaside A and arctiin which existed in the the composition with largerchemical polarity caused obvious pseudoallergic reactions. SHLI with removal forsythoside A with the technology of HPLC-MS displayed reduced pseudoallergic reaction and a significant improved safety. This study provided a scientific basis for SHLI process improvements and also offered idea and research foundation for screening pseudoallergenic substances injections in other TCM injections.
Animals ; Drug Hypersensitivity ; etiology ; Drugs, Chinese Herbal ; adverse effects ; analysis ; Furans ; adverse effects ; Glucosides ; adverse effects ; Glycosides ; adverse effects ; Injections ; Male ; Mice ; Mice, Inbred ICR

OBJECTIVETo evaluate the clinical effect of surgical resection of the severe heterotopic ossification (HO) after the open reduction internal fixation (ORIF) of acetabular fractures.

METHODSFive cases of severe HO after the ORIF of acetabular fractures were treated by surgical resection from October 2005 to April 2007. All patients were male, the average age was 34 years (22 to 45 years). The average time of HO after ORIF of acetabular fractures was 14.2 months (3 to 30 months). The original surgical approaches were: Kocher-Langenbeck approach as 4, ilioinguinal combined K-L approach as 1. According to the Brooker classification, there were 4 patients with IV degree and 1 with III degree. The average total movement for all the 5 patients was 8 degrees. All patients received one time radiation therapy before or after operation, the dosage was 7-8 Gy. The surgical approach was Kocher-Langenbeck for all patients. During operation the nerve stimulator was used to explore the sciatic nerve and carefully protected it, resected all HO bone and removed all implants. For one patient, because of confusion between femoral head and acetabulum, total hip replacement were performed. The joint exercise (passively and actively) began from the second day after operation, and at the same time, all patients took the indomethacin to prevent the occurrence of HO.

RESULTSAll patients were followed up for 4 to 22 months. There was no recurrence of HO, the average total movement for all the 5 patients was 160 degrees.

CONCLUSIONEarly surgical resection and combined with radiation and indomethacin for the severe HO after the ORIF of acetabular fractures can obtain excellent results.

Acetabulum ; injuries ; Adult ; Follow-Up Studies ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Ossification, Heterotopic ; etiology ; surgery ; Postoperative Complications ; surgery ; Treatment Outcome