Objective To evaluate autologous buccal mucosal patch grafting for correcting penile deformity in the surgical treatment of Peyronie's disease. Methods 14 patients with symptoms of Peyronie's disease for more than three months suffered from persistent pain ,severe curvature of the penile shaft and resistant to conservative treatment underwent plaque incision and autologous buccal mucosa patch grafting. Results Postoperative follow up has been 0.5 to 2years.The penile shaft became straight with no narrowing or indentation in 12 patients (86%) . In 14% of cases (2/14) some curvature of the penile shaft persisted and there was pain on erection.In all the patients there was no change in penile length. 86% of patients had overall satisfaction after surgery with improvement in psychological and social well-being. Conclusions The autologous buccal mucosal patch graft provides an anatomical and functional tunied substitute resulting in correction of the penile deformity in Peyronie's disease.

Objective To review the diagnosis and treatment of extra-adrenal pheochromocytoma with a 34cases report. Methods Thirty-four cases of extra-adrenal pheochromocytoma were retrospectively analyzed. Hypertension was observed in 27 cases. Abdominal pain was seen in 10 patients and intermittent hematuria in 2 patients. Serum and urinary catecholamine and urinary VMA were measured in 34 cases. The level of serum or urinary catecholamine elevated in 20 cases and urine VMA elevated in 24 cases. Thirty-four cases had ultrasound examination,25 cases underwent CT scan and 6 cases underwent MER scan.Results Pheochromocytomas of 12 cases were located in the renal hilum, 2 in the lower pole of the left kidney, 1 in the posterior aspect of the inferior vena cava, 3 in the interaortocaval region, 2 in the anterior aspect of the abdominal aorta, 1 in the anterior of the right common iliac artery, 1 in the hilum of the liver, 1 in the posterior o{ the pancreas, 2 in the bladder wall, 1 in the posterior of the descending colon, and 8 cases of multifoci. Twenty-two cases of extraadrenal pheochromocytoma were benign and 12 cases were malignant. Thirty cases were followed up from 6 months to 13 years. Among 27 cases with hypertension, the blood pressure of 22 patients returned to normal and 5 cases were still hypertensive. Nine cases of malignant pheochromocytoma all had tumor recurrence or metastases at one year postoperatively. Six patients died during followed-up from 6 months to 3 years, including 3 cases died of cerebral hemorrhage and 3 cases of tumor metastases. Three cases got stable with 131Ⅰ-MIBG radiotheraphy.Conclusions The accurate detecting extra-adrenal pheochromocytoma is difficult. CT scan could be reliable in localizing the lesions. Surgical resection of the tumor could be the best therapy. Patients of malignant extra-adrenal pheochromocytoma may be treated with 131Ⅰ-MIBG after surgical therapy.

The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.

Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.

OBJECTIVETo investigate the inhibitory effects of OMT on TGF-β1-induced CFBs proliferation, and then explore the mechanism.

METHODThe experiment was randomly divided into 6 groups as following: control group (serum free DMEM), model group (20 μg x L(-1) TGF-β1), OMT low dose group (1.89 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT medium dose group (3.78 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), OMT high dose group (7.56 x 10(-4) mol x L(-1) + 20 μg x L(-1) TGF-β1), SB203580 group (p38MAPK blocking agent, 1 x 10(-5) mol x L(-1) + 20 μg x L(-1) TGF-β1). Vimentin of CFBs was identified by immunocytochemical methods, α-SMA of myFBs as well. Inhibitory effects of OMT on CFBs proliferation was detected by the MTT assay. Picric acid Sirius red staining was analyzed collagen type I and collagen type III deposition. Western blot was determined the expression of p38MAPK, p-p38MAPK, collagen type I and collagen type III.

RESULTMTT results showed that OMT significantly inhibited CFBs proliferation induced by TGF-β1 (P < 0.01) α-SMA immunocytochemical experiments suggested that OMT could protect against the CFBs proliferation. OMT could significantly decrease the deposition of collagen type I and collagen type III by Western bloting and picric acid Sirius red staining. Western blot results showed that TGF-β1 enhanced p38MAPK phosphorylation, however OMT attenuated the phosphorylation of p38MAPK induced by TGF-β1 (P < 0.01).

CONCLUSIONOMT can inhibit the CFBs proliferation induced by TGF-β1, and its mechanism may be involved in inhibiting p38MAPK phosphorylation.

Alkaloids ; pharmacology ; Animals ; Cell Proliferation ; drug effects ; Collagen ; metabolism ; Down-Regulation ; Female ; Fibroblasts ; drug effects ; Heart ; drug effects ; In Vitro Techniques ; Male ; Phosphorylation ; Quinolizines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; antagonists & inhibitors ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo evaluate the expression of nitric oxide synthase III (NOS III) mRNA in the heart, aorta, kidney and liver of spontaneously hypertensive rats (SHR).

METHODSTwo hundred and ninety-four total RNA samples were obtained from the tissues of ventricle, aortic smooth muscle, kidney and liver of SHR and normotensive rats (Wistar-Kyoto rats, WKY). RNA array was used to determine the mRNA levels of NOS III of the two groups.

RESULTSCompared with WKY, the systolic blood pressure increased significantly in SHR at 6-week-old, 8-week-old, 10-week-old and 12-week-old [(158.50 +/-7.69 vs 108.67 +/-5.89) mmHg, (174.33 +/-4.46 vs 128.50 +/-4.97) mmHg, (198.00 +/-13.45 vs 142.00 +/-3.58) mmHg, (216.67 +/-8.91 vs 141.17 +/-4.92) mmHg, P<0.01], and the ventricle/body weight ratio was significant higher at 10-week-old and 12-week-old [(4.08 +/-0.17 vs 3.59 +/-0.11, 4.05 +/-0.18 vs 3.40 +/-0.19)mg/g, P<0.01]. In the heart tissue and the kidney, the mRNA levels of NOS III were significantly increased at 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.12 +/-0.18 vs 0.90 +/- 0.15, 1.46 +/- 0.34 vs 1.06 +/-0.18, 1.66 +/- 0.31 vs 1.21 +/- 0.30, 1.98 +/- 0.40 vs 1.31 +/-0.38, P <0.05) and at 4-week-old, 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.10 +/- 0.21 vs 0.81 +/-0.11, 1.28 +/-0.18 vs 0.95 +/-0.13,1.31 +/-0.23 vs 0.99 +/-0.23, 1.70 +/-0.30 vs 1.08 +/-0.25, 1.83 +/-0.33 vs 1.15 +/-0.20, P<0.05 or P<0.01), respectively. There was no significant difference of the NOS III expression in the liver and no significant signals were detected in the aortic smooth muscle.

CONCLUSIONThe results provide the evidence of the increased expression of NOS III in different tissues in SHR and suggests the progressive nature of essential hypertension.

Animals ; Hypertension ; enzymology ; genetics ; Kidney ; enzymology ; Liver ; enzymology ; Male ; Myocardium ; enzymology ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type III ; Oligonucleotide Array Sequence Analysis ; methods ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

OBJECTIVETo study the main active constituents of Siwu mixture for reversing the multidrug resistance (MDR) of human erythrocyte leukemic cell line K562/ADM.

METHODMacroporous resin was used to separate Siwu mixture and an in vitro model was used to evaluate the reversal index of each fraction for discovery of active fraction. HPLC-MS was used for investigating the main active constituents of Siwu mixture.

RESULTThe reversal index of the gross polysaccharide of Rehmannia glutinosa was significantly higher than that of blank control (P < 0.01). There was little difference between the reversal effect of the gross polysaccharide of Rehmannia glutinosa and that of the active fraction A (P > 0.05).

CONCLUSIONThe gross polysaccharide of Rehmannia glutinosa is one of the main active constituents of Siwu mixture.

Doxorubicin ; Drug Combinations ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; K562 Cells ; Ligusticum ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Rehmannia ; chemistry

The aim of this study is to develop monoclonal antibody against human hepatocyte growth factor activator inhibitor 1 (HAI-1) for future study of HAI-1. The cDNA fragments of human hepatocyte growth factor activator inhibitor 1 (HAI-1) were subcloned to construct GST-HAI-1 fusion protein expression vectors. The vectors were transformed into E. coli and fusion protein expression was induced by IPTG. The GST-HAI-1 fusion proteins were separated on preparative SDS-PAGE and recovered by electroelution, and used to immunize BALB/c mice. Hybridomas producing monoclonal antibodies against human HAI-1 were prepared by cell fusion technique and characterized by ELISA, Western Blot and immunohistochemical staining. One hybridoma cell line, ZMC6, was obtained, which produces specific antibody against the expressed GST-HAI-1 fusion protein. The monoclonal antibody recognizes both the membrane-type and secretory-type HAI-1 proteins of colorectal tissue. The successful development of anti-HAI-1 antibody provides a powerful tool for further investigation on HAI-1's function.
Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Glutathione Transferase ; genetics ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Proteinase Inhibitory Proteins, Secretory ; analysis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; immunology

Thermosensitive gel with the property of LCST (Lower Critical Solution Temperature) was studied as a hot spot these years for it can be used as one supportor of drug controlled release system which can release drug at right point, right time and right quantity. With further reseaches and applications of thermosensitive gel, the modes and mechanisms of it were discovered and poited out, drug release models were also established step by step. Resesrches on drug release mechanisms and modes of thermosensitive gel will make it applied better in fields of medicine and biology.
Drug Delivery Systems ; methods ; Gels ; Models, Theoretical ; Temperature

OBJECTIVEThis study aimed to investigate the effect of trichloroethylene (TCE) intake via drinking water on Th17 cells in mice.

METHODSForty eight six weeks old female BALB/c mice were divided into blank control, vehicle control, 2.5 mg/ml TCE and 5.0 mg/ml TCE groups by random number table (12 mice each group), and exposed to TCE by drinking water. On the 14(th), 28(th), 56(th), 84(th) days, blood were collected and assayed for IL-17, IL-6, and TGF-β concentration in serum through ELISA. Animals were killed and spleen biopsies were taken sterility. The proportion of Th17 cells among CD4(+) T cells and RORγt mRNA expression level in spleen were measured by FCM and real-time PCR.

RESULTSIn 2.5 mg/ml TCE and 5.0 mg/ml TCE group mice, Th17 cells/CD4(+) T cells in spleen were (3.46 ± 0.32)% and (5.45 ± 0.45)% on day 14, (3.47 ± 0.33)% and (4.10 ± 0.39)% on day 84, which were significantly higher than those for solvent control group at the same time point ((2.15 ± 0.20)%, (2.16 ± 0.35)%, respectively) (P < 0.01). RORγt mRNA expression levels were (1.870 ± 0.084) and (1.965 ± 0.060) on 14 day, (1.998 ± 0.079) and (2.028 ± 0.073) on day 56, which were also significantly higher than those for solvent control group at the same time point (1.77 ± 0.04 and 1.75 ± 0.09, respectively) (P < 0.05). IL-17 concentrations in serum were (32.28 ± 5.38) and (34.47 ± 5.02) pg/ml on day 14, and (34.87 ± 5.48) and (41.94 ± 6.19) pg/ml on day 28, which were significantly higher than those for solvent control group at the same time point((21.57 ± 5.23), (22.11 ± 5.11) pg/ml). IL-6 concentration in serum were (43.07 ± 6.71) and (47.86 ± 8.52) pg/ml on day14, (41.32 ± 7.04) and (46.74 ± 9.33) pg/ml on day 56, which were significantly higher than solvent control group at the same time point ((7.56 ± 7.71) and (28.26 ± 7.22) pg/ml). TGF-β concentration were (17.48 ± 3.06) and (18.93 ± 3.12) pg/ml on day 14, which did not show significant difference from solvent control group ((15.25 ± 2.95) pg/ml). Correlation analysis showed that IL-6 in serum were significantly positively correlated with the proportion of Th17 cells among CD4(+) T cells and RORγt expression level in spleen (r = 0.741, 0.765, P < 0.01).

CONCLUSIONTCE might promote the differentiation of Th17 cells and increase IL-17 secretion by inducing IL-6 and up-regulating RORγt expression together with TGF-β.

Animals ; Drinking Water ; chemistry ; Female ; Interleukin-17 ; immunology ; Interleukin-6 ; immunology ; Mice ; Mice, Inbred BALB C ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; immunology ; Th17 Cells ; drug effects ; immunology ; Transforming Growth Factor beta ; immunology ; Trichloroethylene ; toxicity