Objective Radionuclide-labeled low molecular weight polypeptide is reeently advocated for the diagnosis and treatment of malignant tumor. The purpose of this study was to evaluate the anti-tumor effect of 131Ⅰ-Tyr-octreotide in nude mice bearing human non-small cell lung cancer (NSCLC). Methods 131Ⅰ-Tyr-octreotide was prepared by Ch-T method. The radiochemical purity was measured and biodistribution was evaluated. The nude mice models bearing human NSCLC were studied and divided into four groups: group A injected 131Ⅰ-Tyr-octreotide through tail vein, group B injected normal saline, group C injected 131Ⅰ-Tyroctreotide through stroma and group D injected 131Ⅰ through stroma. The radioactivity ratio of tumor to normal tissue (T/NT) was calculated over region of interest (ROI). The tumor cell cycle and cell apoptosis were analyzed by flow cytometry (FCM), terminal deoxynucleotidyl transferase mediated dUTP-biotion nick end labeling (TUNEL) and histopathological analysis. Statistical analysis was performed with SPSS 11.0, and the comparison for difference between groups performed with one-way ANOVA analysis. Results The labeled radiochemical purity was (95.23±1.67)% and specific activity of 3.5×106Bq/ug. The biodistributiou showed high uptake in kidney, and low uptake in liver and spleen. The radioactive uptake in group C was higher than the other groups, and the retention time was longer. The T/NT was 52.74±0.13 after 24 h, which was much higher than that the other groups (group D: 8.90±0.23, group A: 6.42±0.02, q=628.81 and 664.33, all P<0.05). The resuits of tmnor cell cycle determined by FCM showed that the G1 phase was blocked mast remarkably in group C than the other groups [group C: (83.17±6.86)%, group A: (57.02±18.81)%, group D: (49.29±7.80)%, group B: (45.88±5.13)%, q=5.29, 6.86, 7.55, 1.56, 2.26, 0.69, all P<0.05]. Apeptotic cells were observed by TUNEL, and apoptotic body was detected by immuno-histochemical examination. Conclusions 131Ⅰ-Tyr-octreotide was easily labeled by Ch-T. 131Ⅰ-Tyr-octreotide could induce tumor cell apoptosis and inhibit the tumor cell of NSCLC. It might be a potential target-directed agent in NSCLC.

Objective To provide an experimental basis for long-term in vitro proliferation,tissue construction and function maintaining of human primary hepatocytes by preparing three-dimensional (3D) porous specific extracellular matrix (ECM) derived from porcine liver with different decellularizing methods.Methods The porcine liver tissue slices were treated using six different decellularization/oxidation ways,and the decellularization extent and ECM structure were evaluated through qualitative and quantitative analysis.Results The decellularized liver-ECM could maintain the original shape during the whole process of the decellularization.HE staining showed that the cellular components and nucleus disappeared in the decellularized ECM,indicating that all of these six decellularization methods can completely decellularize the porcine liver tissue slices.Determination of DNA content showed that 91％ ～ 97％ of cell components have been removed.Electron microscopy scanning observed that the pore sizes and ultrastructures of the liver ECMs under six decellularization treatments were significantly different.Conclsion The six decellularization/oxidation methods evaluated in this study could successfully create a 3D specific decellularized liver ECM with porous ultrastructure,which further lays basis for the development of a novel in vitro 3D culture model for human hepatocytes.

Objective To observe the effects of simplified recipe ofBuyang Huanwu Decoction on expression of APJ in the brain of local cerebral ischemia rats;To discuss its mechanism of action. MethodsFocal cerebral ischemia rat models were established by middle cerebral arterial occlusion. The adult rats were randomly divided into sham-operation group, model group,Buyang Huanwu Decoction group and simplified recipe ofBuyang Huanwu Decoction group. Administration groups were given relevant medicine for gavage. The expressions of APJ protein and APJ mRNA at different time points were detected by Western blot and RT-PCR.ResultsCompared with model group and sham-operation group, the expression of APJ protein and APJ mRNA at different time points in Buyang Huanwu Decoction group and simplified recipe ofBuyang Huanwu Decoction group significantly increased (P<0.05). The expression of APJ protein at different time points showed no difference between simplified recipe ofBuyang Huanwu Decoction group andBuyang Huanwu Decoction group;while the expression of APJ mRNA in simplified recipe ofBuyang Huanwu Decoction group was higher than Buyang Huanwu Decoction group (P<0.05).ConclusionSimplified recipe ofBuyang Huanwu Decoction plays a role in neural protection and restoration by promoting the expression of APJ in the brain of focal cerebral ischemia rats.

Objective:To analyze the changes in T lymphocyte subsets, B lymphocytes and NK cells in children with active tuberculosis (TB) and their clinical significance.Methods:T lymphocyte subsets, B lymphocytes and NK cells in peripheral blood samples of 106 patients with acute TB (TB group) and 106 healthy children (healthy control group) were detected by flow cytometry and compared between different groups.Results:The percentages of CD3 + T, CD4 + T and NK cells as well as the CD4 +/CD8 + T cell ratio were significantly lower in the TB group than in the healthy control group ( Z=-3.783, P=0.000; Z=-5.401, P=0.000; Z=-3.434, P=0.001; Z=-2.014, P=0.044). The percentages of double negative T (DNT) and B cells in the TB group were significantly higher than those in the healthy control group ( Z=2.765, P=0.006; Z=6.880, P=0.000). No significant difference in the percentage of CD8 + T or double positive T (DPT) cells was observed between the two groups ( P>0.05). The expression of peripheral lymphocyte subsets varied in TB children of different age groups (0-<3, 3-<6, 6-<10 and 10-<16 years old). There were significant differences in CD3 + T, DNT and B cells among the four age groups ( H=10.081, P=0.018; H=14.583, P=0.002; H=8.498, P=0.037). The percentage of CD4 + T cells was significantly lower in children with extrapulmonary TB than in those with pulmonary TB ( Z=-3.068, P=0.002). No statistically significant difference in other lymphocyte subsets was found between children with extrapulmonary and pulmonary TB ( P>0.05). Conclusions:Tuberculosis could lead to immune dysfunction in children. Dynamic monitoring of the changes in peripheral lymphocyte subsets in children with TB could be conducive to better assessment of immune status and providing personalized treatment.

Objective To study the doses and methods of F1 antigen(F1Ag) to immune Balb/c mice during the establishment of hybridoma cell strains. Secreting McAbs against F1Ag of Yersinia pestis. Methods Balb/c mice of seven to nine weeks old were randomly divided into six groups. The first four groups were 150, 100, 50 and 25 μg F1Ag inoculated group, having multipoint hypodermic inoculation of F1Ag of 150, 100, 50 and 25 μg followed by multipoint hypodermic inoculation of F1Ag of 100 μg for a second time and then intraperitoneal injection of 100 μg. Next, hypodermically inoculated group received F1Ag of 100 μg for three times in multiple points. Finally, the intraperitoneal injection group was intraperitoneally inoculated with F1Ag of 100 μg for three times. Emulsification liquid of F1Ag + Complete Frednd's adjuvant(CFA) of equivalence was used in the first inoculation, emulsification liquid of F1Ag + Incomplete Frednd's adjuvant(IFA) balanced mix in the second, F1Ag liquid in the third. One week afterwards, tail blood of the mice was collected to test antibody titers of anti-F1Ag by double antigens sandwich enzyme linked immunosorbent assay (DAgS-ELISA) and trace indirect hemagglutination assay(IHA). Results The levels of antibody of anti-F1Ag in 150,100,50 and 25 μg groups had statistics difference (DAgS-ELISA method: G = 12 173.87,13 440.37,15 024.19 and 4466.72, F= 3.11, P< 0.05;IHA: G = 19 972.32,18 089.40,23 170.47 and 4871.08, F = 4.11, P < 0.05). Immune effect of the 3 groups of 150, 100 and 50 μg was almost the same (P> 0.05), and excelled as compared with that in 25 μg group with statistics difference(DAgS-ELISA method: t = 2.18,2.39,2.73, P < 0.05;IHA: t = 2.54,2.73,3.13, P< 0.05). The titer of F1 antibody had an increasing trend from the 100 μg group to hypodermic group and intraperitoneal injection groups, but without statistics difference (DAgS-ELISA method: G = 8933.44, 9986.16, 13 440.37;IHA: G = 13 777.25,16 384.00, 18 089.40, F = 0.66,0.25, all P > 0.05). Conclusions Hyodermical inoculation of F1Ag with the first dose of 50 μg in multiple points for mouse is appropriate, and a strengthening dose of 100 μg in an intraperitoneal injection may shorten the immune period.

Objective To study the sensitivity and specificity of a double monoclonal antibody sandwich enzyme-linked immunosorbent assay (DMcAbS-ELISA)for the detection of F1 antigen of Yersinia pestis (Y.pestis).Methods Viscera (viz.liver and spleen)specimens of infected mice with virulent Y.pestis and negative control mice were detected by bacteriological test,DMcAbS-ELISA and reverse indirect hemagglutination assay (RIHA) for the F1 antigen.Results The 225 control specimens were all negative tested by plague bacteriology testing,DMcAbS-ELISA and RIHA.A total of 308 plague-infected mouse organ specimens were tested,and the positive detection rate was 92.21％ (284/308),90.91％(280/308) and 89.61％ (276/308),respectively,with germiculture,DMcAbS-ELISA and RIHA,and the difference was not statistically significant(x2=5.65,P＞0.05).The coincidence rate of DMcAbS-ELISA and bacterial culture was 97.00％[(274+243)/533],Kappa =0.940;RIHA in line with the rate was 99.25％[(276+253)/533],Kappa =0.985.Authenticity comparison of F1 antigen detection in viscera specimens:sensitivity,specificity,positive predictive value,negative predictive value,adjusted agreement and Youden's index was 96.48％(274/284),97.59％(243/249),97.86％ (274/280),96.05 ％(243/253),96.99％[1/4×(274/280+274/284+243/253+243/249)]and 0.9407,respectively,for DMcAbS-ELISA and 96.13％(273/284),98.80％(246/249),98.91％(273/276),95.72％(246/257),97.39％[1/4×(273/276+273/284+246/257±246/249)]and 0.9492,respectively,for RIHA.The detection sensitivity of DMcAbS-ELISA and RIHA was 2.7×104 cfu/ml and 2.2×105 cfu/ml,for Y.pestis,respectively,and was 10 μg/L for F1 antigen.Conclusions DMcAbS-ELISA assay is a sensitive,specific,simple and fast method for detection of the F1 antigen,and it has a potential application value in rapid diagnosis of plague.

AIMTo observe the changes of MMP-2, 9 level on atherosclerosis in experimental rats by treatment of 2,3,4',5-tetrahydroxystilbene-2-0-beta-D glucoside (TSG) and to investigate the mechanism of TSG in stabilizing plaque and anti-atherosclerosis.

METHODSThe atherosclerosis model of rat was made by feeding high grease food and injecting VitD3. Sixty male SD rats were randomly divided into six groups: control, Simvastatin, model and TSG 120 mg x kg(-1) x d(-1), TSG 60 mg x kg(-1) x d(-1) and TSG 30 mg x kg(-1) x d(-1). After 12 weeks, several aorta were randomly tested, model and TSG 120 mg x kg(-1) x d(-1), TSG 60 mg kg(-1) x d(-1) and TSG 30 mg x kg(-1) x d(-1). After 12 weeks, several aorta were randomly tested, model made was successful when we found plaque. And after six weeks treating, the mRNA expressions of MMP-2 and MMP-9 were measured by RT-PCR. The activities of MMP-2 and MMP-9 were measured by Western blot. The levels of CRP, IL-6 and TNF-alpha in serum were measured in biochemical method.

RESULTSData of the study demonstrated that the level of TNF-alpha, IL-6, CRP, MMP-2 and MMP-9 were remarkably decreased by TSG60, 120 mg x kg(-1) x d(-1) groups, which showed a dose-dependent effect.

CONCLUSIONTSG has the effect of anti-atherogenic and stabilizing plaque on the experimental rats with atherosclerosis, which are induced by the high cholesterol feeding and VitD3 injecting. The effect of TSG seems to be closely involved in regulating the expressions of MMP-2 and MMP-9, and inhibiting inflammation.

Animals ; Atherosclerosis ; drug therapy ; metabolism ; C-Reactive Protein ; metabolism ; Glucosides ; isolation & purification ; therapeutic use ; Interferon-alpha ; blood ; Interleukin-6 ; blood ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Polygonum ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; isolation & purification ; therapeutic use

OBJECTIVETo investigate the clinical effect of microsurgical vasoepididymostomy and/or vasovasostomy in the treatment of obstructive azoospermia.

METHODSThis study included 76 patients with obstructive azoospermia, 53 treated by bilateral vasoepididymostomy (8 involving the epididymal head, 18 involving the epididymal body, 5 involving the epididymal tail, and 22 involving the epididymal head, body and tail), 14 by unilateral vasoepididymostomy, and the other 9 by unilateral vasoepididymostomy + unilateral vasovasostomy (including cross anastomosis). We followed up the patients for 2 to 16 months for the patency rate, routine semen parameters, and pregnancy outcomes.

RESULTSThe success rate of bilateral vasoepididymostomy, unilateral vasoepididymostomy, and unilateral vasoepididymostomy + unilateral vasovasostomy (including cross anastomosis) were 62.26% (33/53), 35.71% (5/14), and 77.78% (7/9), respectively. The average sperm concentrations in the three groups of patients were (27.9 +/- 5.74), (11.8 +/- 8.33), and (19.9 +/- 7.53) x 10(6)/ml, the average total sperm counts were (65.6 +/- 13.71), (28.0 +/- 15.86), and (69.2 +/- 28.59) x 10(6), and the mean rates of progressively motile sperm were (22.3 +/- 3.18), (11.0 +/- 9.77), and (15.8 +/- 5.05)%, respectively. The success rates of bilateral vasoepididymostomy that involved the epididymal head, body, tail, and all the three parts were 62.5, 72.22, 60, and 54.55%, respectively. Natural pregnancy was achieved in 8 (10.53%) of the total number of cases.

CONCLUSIONMicrosurgery is effective for the treatment obstructive azoospermia. Unilateral vasoepididymostomy + unilateral vasovasostomy is superior to the other procedures, followed by bilateral vasoepididymostomy. Bilateral vasoepididymostomy involving the epididymal body may achieve a slightly better effect than that involving the other epididymal parts.

Adult ; Anastomosis, Surgical ; methods ; Azoospermia ; etiology ; surgery ; Epididymis ; surgery ; Female ; Humans ; Infertility, Male ; surgery ; Male ; Microsurgery ; Pregnancy ; Pregnancy Rate ; Sperm Count ; Treatment Outcome ; Vas Deferens ; surgery ; Vasovasostomy ; methods

5 mm and ≤8 mm in 4 cases.The mean value was 4.2 mm. Four patients noticed reduction in their vision and two had diplopia.Those patients were examined by CT or MR.Direct venography was performed in each patient.After the diagnosis of OVM was confirmed, intralesional injection of BLE was performed.The efficacy of the treatment and complications were observed during the following 8 to 42 months(mean 23 months).Results The BLE were successfully injected in all the patients.All patients had resolution of proptosis and diplopia.Three patients gained improvement of visual acuity.The periorbital swelling occurred in all patients after operation and resolved within 1 week without special treatment.Other complications,such as orbital hemorrhage and periorbital scar,were not observed during following-up.Conclusion Intralesional injection with BLE is convenient,safe and efficient for the treatment of OVM.

Objective To study the sensitivity and specificity of gold-immunochromatography assay (GICA) for detection of Yersiniapestis(Y. pestis ) F1 antigen. Methods Viscera organ(liver and spleen) specimens of 308 mice with virulent Y. pestis infection and 225 control specimens of rats(217 Spermophilus dauricus, 5 mice,3 guinea pigs) were detected by GICA dipstick with monoclonal antibody against plague F1 antigen (F1MAb).Meanwhile, micro-method of reverse indirect hemagglutination assay(RIHA) and bacteria culture were carried out for parallel testing. Results Bacteriological examination of 225 control specimens, and F1 antigen detected with GICA and RIHA were all negative. No cross-reaction with related Yersinia pseudotuberculosis at 1 x 108 cfu/ml level was found in GICA and RIHA. Detection sensitivity of Y. pestis by GICA and RIHA were 2.5 × 105 cfu/ml and 2.0 × 105 cfu/ml, respectively, and of F1 antigen were 1μg/L and 10 μg/L, respectively. Coincidence was 97.94% (522/533) between GICA and bacteriological test, Kappa = 0.959, and the difference was statistically insignificant(x2 = 0.36, P ＞ 0.05); and 97.94%(522/533) between GICA and RIHA, Kappa = 0.959, with statistically significant difference in the positive detection rates(x2 = 9.09, P ＜ 0.05). Out of the 308 infected mice, 284 were positive of plague bacterial cultured, In 284 samples with positive bacterial culture, there were 280 of positive detected by GICA for F1 antigen, positive rate of F1 antigen was 98.59%, higher than that by RIHA[the positive rate of 96.13%(522/533)], with statistically significant difference(x2 = 5.14, P ＜ 0.05). Sensitivity of GICA was 98.59% (280/284), specificity was 97.19% (242/249), positive predictive value (PPV) was 97.56% (280/287),negative predictive value ( NPV ) was 98.37% (242/246), and Youden index was 0.9578. Conclusions GICA is sensitive and specific, fast and simple in detection of F1 antigen of the plague. It's a valuable detection technique for early and rapid diagnosis of plague.