10.3969/j.issn.1000-3606.2013.06.016

Objective To prepare the mouse anti recombinant human Galectin-7 antibody and the antibody was characterized in bladder cancer.Methods The gene coding for Galectin-7 was amplified by PCR from the cDNA of human foreskin cells and cloned into prokaryotic expression vector pET28a.Then the recombinant plasmid pET28a/Galectin-7 was transformed into E.coli BL21 (DE3)and expressed under IPTG induction.The recombinant Galectin-7 was purified through Ni2+-NT agarosegel column and the purified Galectin-7 used as imunogen to imunize the mouse.The titer and specificity of the anti-Galectin-7antibody from the mouse were analyzed by ELISA,Western blot and immunohistochemistry,respectively.Results The recombinant Galectin-7 was successfully expressed and purified,and the polyclonal ani-Galectin-7 antibody was suc-cessfully prepared.The titer of the antiserum was 1∶32 000 by ELISA.Western blot analysis showed this antiboday reacted specifically with Galectin-7.Immunohistochemistry analysis showed the antibody could recognize the native Galectin-7 in the human bladder cancer tissue.Conclusion The preparation recombinant Galectin-7 protein was as immunogen in rabbits.It was successful to produce high titer and high specificity of anti Galectin-7 polycolonal antibody.

Objectiye To investigate the expression of regulated on activation, normal T-cell expressed and secreted (RANTES) and occluding in brain, and investigate the role of expression of RANTES in the change of blood-brain barrier permeability of rats with acute pancreatitis. Methods 49 Sprague-Dawley rats were randomly divided into 7 groups according to random number table, including sham-operated group (SO) , acute edematous pancreatitis (AEP) 3, 6 h group and acute necrotizing pancreatitis (ANP) 3, 6, 12 and 24 h group. AEP and ANP were induced by retrograde injection of 0. 5% and 5% sodium taurocholate, respectively. RT-PCR, Western blot, and immunohistochemistry methods were performed to assess the expression of RANTES and occludin mRNA and protein in brain tissue. Results The RANTES mRNA expressions in SO, AEP 3, 6 h, ANP 3, 6, 12, 24 h group were 0, 0, 0, 0.36±0.05, 0.47±0.04, 0.65±0.05, 0.83±0.07, respectively; The RANTES protein expressions were 0, 0, 0, 0.42±0. 03, 0. 57±0.04,0.78±0.08, 1.05±0.08, respectively; the values in ANP group were significantly lower than those in SO group and AEP group (P<0.01). The occludin mRNA expressions were 1.21±0.07,1.17±0.07, 1.15±0.08,0.84±0.07,0.77±0.05,0.64±0.09,0.56±0.09, respectively, the occludin protein expressions were 1.18 ±0.08, 1. 16 ±0. 10, 1. 11 ±0. 10, 0. 90 ±0. 03, 0. 65 ±0."05, 0.57 ±0.05, 0.48 ±0.05, respectively, the values in ANP group were significantly lower than those in SO group and AEP group (P< 0.01). The expression of RANTES was positively related to the pancreatic pathologic score (r = 0. 936,P< 0. 001) ; the expression of occluding was negatively related to the pancreatic pathologic score (r = - 0. 943, P < 0.001). The expression of RANTES was negatively related to the expression of occluding (r = -0. 943, P <0. 001). Conclusions The expression of RANTES was progressively increased in the brain tissue in rats with ANP, while the expression of occluding was progressively decreased; RANTES may play an important role in the change of blood-brain barrier permeability via down-regulating the expression of occluding.

Objective To compare the clinical and radiological outcomes between hybrid and total pedicle screw instrumentation in adolescents undergone posterior spinal fusion (PSF) for thoracic scoliosis secondary to Chiari malformation..Methods A total of 75 patients undergone PSF were included and divided into two groups:the all pedicle screw group (Group A,n=44) and the hybrid group (Group B,n=31).Patients were evaluated before surgery,immediately after surgery,and at the 2-year follow-up in radiographic changes in curve magnitude,apical vertebral translation (AVT),apical vertebral rotation (AVR),trunk shift,thoracic kyphosis (TK),lumbar lordosis (LL),and sagittal vertical axis (SVA).These parameters were further analyzed with respect to preoperative TK in both groups.Results After surgery,the average correction of the thoracic curve was 60.2％ and 51.3％ in Group A and B,respectively (t=2.372,P=0.023).The average lumbar curve correction was 61.7％ in Group A,representing a significant increase compared to Group B (51.1％,t=2.431,P=0.020).At the final follow-up,loss of the thoracic curve correction was less in Group A (0.3％) than in Group B (1.7％),however,there was no statistical significance (t=-0.468,P＞0.05).AVT of the thoracic curve improved in Group A from 25.0 mm to 6.9 mm,while in Group B it changed from 24.1 mm to 7.4 mm.For patients with a preoperative TK greater than 40°,the proximal junctional angle was found to be significantly larger in Group A (10.0 degrees versus 4.5 degrees,t=-2.031,P=0.052) by the final follow-up,along with a significantly increased incidence of proximal junctional kyphosis (20％ versus 9％).Conclusion Total pedicle screw instrumentation provided a significantly better correction of the major and minor curves than hybrid constructs for the operative treatment of thoracic scoliosis secondary to Chiari malformation.However,for patients with thoracic hyperkyphosis,all-screw instrumentation had a higher risk of adjacent level proximal kyphosis.

This editorial briefly reviewed the history of andrology, its rapid development in recent years, the conception of male health, international andrology studies, the achievements of Chinese andrologists and the existing problems. It is highly necessary for Chinese andrology community to strengthen the international academic communication, join the International Society of Andrology and attend its activities actively.
Andrology ; trends ; Biomedical Research ; trends ; China ; Humans ; Male

OBJECTIVETo investigate the genotype of blaADC which was a kind of AmpC produced by Acinetobacter baumannii (AB), isolated through the detection of 28 similar strains among children.

METHODS28 strains of AB were collected and isolated from the Pediatrics clinic during 2006, and were identified through bacteria and susceptibility test using Vitex-32 automicroscan GNI and GNS cards. The genotype of blaADC was confirmed by polymerase chain reaction (PCR) and them sequenced.

RESULTS3 of the 28 strains of AB showed multi-drugs resistance, with a positive rate of 10.71%. blaADC was discovered in 17 of the 28 strains and the positive rate was 60.71%. All the 28 strains of AB were resistant to Cefoxitin. blaADC positive strains were all sensitive to Ampicil/Sulbactam, and only one of them was resistant to Piperacillin/Tazobactan. There were no blaADC genes discovered in the strains that were resistant to Ampicil/Sulbactam or Piperacillin/Tazobactan. There were changes of amino acids on the site 4, 242, 342 and 376 in the sequence of blaADC of No.2 strain, comparing to gi /7258342/ emb /CAB77444. 1/ in GenBank.

CONCLUSIONAbove 60% of the AB isolated in children were carrying blaADC while a strain was collected from them at random. When they were undertaken nucleotide sequence analysis, significant difference was found from the others that landed in GenBank, which identified itself as new subtype.

Acinetobacter Infections ; microbiology ; Acinetobacter baumannii ; classification ; drug effects ; genetics ; isolation & purification ; Amino Acid Sequence ; Anti-Bacterial Agents ; pharmacology ; Bacterial Typing Techniques ; Child ; DNA, Bacterial ; genetics ; Drug Resistance, Multiple, Bacterial ; genetics ; Genotype ; Humans ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Sequence Alignment ; Sequence Analysis, DNA ; beta-Lactamases ; genetics

In order to evaluate the effect and mechanism of the mulberry leaf alkaloid, flavones, and polysaccharide intervention on diabetes, the overall metabolite profiling characteristics for the plasma of diabetic mouse was performed by using an ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS). The 8 potential biomarkers were found in diabetic mice plasma based on the data of MS/MS characteristics obtained from the UPLC-OrbitrapMS analysis, which mainly involved in sphingolipids, amino acid metabolic pathway. The principal component analysis showed that the normal group and model group were obviously distinguished and implied that metabolic disturbance was happened in diabetic mice plasma. The extracts of mulberry leaf flavonoids, polysaccharide, alkaloid had exhibited the effects of callback function for diabetic mice through regulating the amino acid metabolism and sphingolipid metabolism.

Objective To evaluate the optical quality in patients with meibomian gland dysfunction (MGD) by using double-pass optical quality analysis system (OQAS-Ⅱ) for offering the reference data for clinical practices.Methods A total of 50 patients with a diagnosis of MGD,who were divided into MGD with dry eye disease (DED) group (n =22) and MGD without DED group (n =28) according to the results of tear film break-up time (BUT) and Schirmer I test (SIT),and another 25 healthy subjects were enrolled in this study.Then several parameters,including modulation transfer function cutoff (MTF cutoff),Strehl ratio (SR) and objective scattering index (OSI),were measured to evaluate the optical quality by using OQAS-Ⅱ.Additional clinical examination,including meibomian gland and tear film assessment were per formed and compared between the three groups.In addition,a correlation analysis was conducted among OSI,MTF cutoff,SR,BUT,SIT and meibomian gland assessment.Results Statistical differences were approached in MTF cutoff,SR,OSI of the three groups (all P ＜ 0.05).The MTF cutoff and SR in MGD without DED group were significantly lower than those in the control group [(31.36 + 1.83) c · deg-1 vs.(35.87 ± 1.59)c·deg-1,(0.21 +0.02) vs.(0.23 ±0.03)],but the OSI got higher [(0.57+ 0.06) vs.(0.45 ±± 0.06)] (all P ＜ 0.05).The SR and MTF cutoff in MGD patients with DED was significantly lower than those in MGD patients without DED [(27.87 ±± 3.08),c·deg-1 vs.(31.36±±1.83)c· deg-1,(0.16 ±±0.02) vs.(0.21 ± 0.02)],but OSIgot significantly higher [(0.72 ± 0.10) vs.(O.57 ± 0.06)] (all P ＜ 0.05).Among the three groups,OSI,MTF cutoff and SR showed no significant correlation with BUT,SIT and meibomian gland assessment (all P ＞ 0.05).Conclusion There is changes in visual quality parameters in MGD patients by OQAS-Ⅱ,and MGD with DED patients has significant changes than MGD patients without DED.

In order to evaluate the effect and mechanism of the mulberry leaf alkaloid, flavones, and polysaccharide intervention on diabetes, the overall metabolite profiling characteristics for the plasma of diabetic mouse was performed by using an ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS). The 8 potential biomarkers were found in diabetic mice plasma based on the data of MS/MS characteristics obtained from the UPLC-OrbitrapMS analysis, which mainly involved in sphingolipids, amino acid metabolic pathway. The principal component analysis showed that the normal group and model group were obviously distinguished and implied that metabolic disturbance was happened in diabetic mice plasma. The extracts of mulberry leaf flavonoids, polysaccharide, alkaloid had exhibited the effects of callback function for diabetic mice through regulating the amino acid metabolism and sphingolipid metabolism.
Alkaloids ; chemistry ; Amino Acids ; metabolism ; Animals ; Biomarkers ; blood ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; drug therapy ; Flavones ; chemistry ; Flavonoids ; chemistry ; Metabolic Networks and Pathways ; Metabolomics ; Mice ; Morus ; chemistry ; Plant Leaves ; chemistry ; Sphingolipids ; metabolism ; Tandem Mass Spectrometry

Objective To evaluate the role of tumor necrosis factor α-induced protein 8-like-2 ( TIPE2) in pyroptosis in macrophage of mice using small interfering RNA ( siRNA) technique. Methods J774A. 1 macrophages of mice were divided into 4 groups ( n=6 each) using a random number table method: non-specific siRNA (Scr-siRNA) group (S group), Scr-siRNA plus LPS∕ATP group (S+LPS∕ATP group ) , TIPE2-siRNA group ( T group ) and TIPE2-siRNA plus LPS∕ATP group ( T+LPS∕ATP group) . The corresponding siRNA and Lipofectamine2000 transfection reagents were added to each group, and transfection was performed for 24-48 h, and in addition LPS 1. 0 μg∕ml was then added, cells were incubated for 5 h, then ATP 5. 0 mmol∕L was added, and cells were incubated for 1 h in S+LPS∕ATP and T+LPS∕ATP groups. Cells were collected to detect the expression of TIPE2, NOD-like receptor familypyrin domain containing 3 (NLRP3), caspase-1, apoptosis-associated speck-like protein containing a caspase activation and recruitment domain ( ASC) and Gasdermin D ( GSDMD) ( by Western blot) . The superna-tant was collected for determination of lactic dehydrogenase ( LDH) activity and concentrations of interleu-kin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results Compared with group S, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, caspase-1, ASC and GSDMD was up-regulated, and the LDH activity and concentrations of IL-1βand IL-18 in supernatant were increased in group S+LPS∕ATP ( P<0. 05) . Compared with group T, the expression of TIPE2 was sig-nificantly down-regulated, the expression of NLRP3, caspase-1, ASC and GSDMD was up-regulated, and the LDH activity and concentrations of IL-1βand IL-18 in supernatant were increased in group T+LPS∕ATP (P<0. 05). Compared with group S+LPS∕ATP, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, caspase-1, ASC and GSDMD was up-regulated, and the LDH activity and con-centrations of IL-1βand IL-18 in supernatant were increased in group T+LPS∕ATP ( P<0. 05) . Conclusion Application of siRNA technique once again confirms that TIPE2 can inhibit pyroptosis in macrophages of mice.