Objective To evaluate the effects of mild hypothermia performed at different times on the levels of glutamate, Bcl-2 and Bax during global cerebral ischemia-reperfsusion (I/R) in rats. Methods Twenty-four male SD rats weighing 230-270 g were randomly divided into 4 groups according to the time at which mild hypothermia was performed ( n =6 each): group A, B, C and D. Global cerebral ischemia was produced by occlusion of 4 vessels (cauterization of bilateral vertebral arteries and occlusion of bilateral common carotid arteries) in the 4 groups. In group B, C and D, the nasopharyngeal temperature was reduced to 32.5-33.5 ℃ and maintained for 1 h. Ischemia was performed after termination of cooling in group B. While ischemia was performed, cooling was started in group C. While reperfusion was performed, cooling was started in group D. Rewarming was started after termination of cooling in group B, C and D. Samples of dialysate in hippocampal CA1 area were collected every 10 min during 100 min reperfusion for determination of glutamate concentrations by high-performance capillary electrophoresis with laser-induced fluorescence detection ( HPCE-LIF). The brain tissues were taken at 3 h of reperfusion for determination of the expression of Bax and Bcl-2 in hippocampal CA1 area, and the ratio of Bcl-2/Bax was calculated. Results Compared with group A, the glutamate concentrations were significantly decreased, the Bcl-2 expression was up-regulated, the Bax expression was down-regulated and the ratio of Bcl-2/Bax was increased in the other thee groups ( P ＜ 0.05). Compared with group B, the Bcl-2 expression was significantly down-regulated, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group C ( P ＜0.05). The glutamate concentrations were significantly increased, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group D compared with group C ( P ＜ 0.05 ). Conclusion The earlier cooling is performed, the better the cerebral protective effect is during global cerebral I/R in rats.

Objective To assess the differential diagnosis and characteristics of renal benign and malignant tumors with three-dimensional contrast-enhanced ultrasound (3D-CEUS). Methods Totally 68 patients with renal tumors were examined by conventional ultrasound and two-dimensional contrastenhanced ultrasound(2D-CEUS). 3D imaging was reconstructed from 2D imaging, the differential diagnosis of renal tumors with 3D-CEUS was analyzed by comparing with 2D-CEUS. All patients with renal tumors were proved by operational pathology. Results Eighteen patients with renal benign tumors mostly displayed equal or low enhancement, showed "slowly in and slowly out" with 2D-CEUS, while displayed regular peripheral and internal vessels with 3D-CEUS. Fifty patients with renal malignant tumors mostly displayed high enhancement, showed "rapidly in and rapidly out" with 2D-CEUS,displayed winding peripheral vessels and disordered internal vessels with 3D-CEUS. 3D-CEUS may display the vascular characteristics of tumors and showed superior imaging quality to 2D-CEUS ( P ＜ 0.05). Conclusions 3D-CEUS can display the vascular characteristics of tumors and their spatial positions, it plays an important role in differential diagnosis between renal benign and malignant tumors.

Objective To explore the value of contrast-enhanced ultrasound(CEUS) in the diagnosis of hematoma and active hemorrhage of renal trauma. Methods Totally 28 patients with renal trauma were examined by conventional ultrasound and CEUS,respectively,including 24 cases caused after renal biopsy,4closed trauma. The detectability of renal hematoma and active hemorrhage with these two methods were compared. All patients were identified by CT or follow-up studies with ultrasound. Results The detectability of renal hematoma with conventional ultrasound and CEUS were 67. 86% (19/28), 92. 86%(26/28), respectively. There was statistically different for detectability in the diagnosis ( P ＜0.05), and the extent of hematoma was more obvious with CEUS. CEUS diagnosed 7 of 26 were renal hematoma with active hemorrhage,which were difficult to be detected with conventional ultrasound. For renal hematoma,the features of CEUS were no enhancement found in every phase; for renal hematoma with active hemorrhage,the contrast agents overflowed from injured blood vessels and formed irregular remarkable enhanced regions. Conclusions CEUS is useful in diagnosing hematoma and identifying the extent and active hemorrhage of renal trauma,in addition,CEUS is valuable in detecting complications after renal biopsy.

Objective To study the X-ray features and differential diagnosis of duodenal obstruction in neonates.Methods X-ray data of duodenal obstruction in 52 cases confirmed surgically were analyzed retrospectively.Erect abdominal plain films were done in each case;Upper gastric intestinal investigations(UGI) were performed in 28 cases,and barium or meglumine diatyizoate enemas were done in 36 cases.Results On the plain films,9 cases with single-bubble,29 with double-bubble,6 with tri-tubble,8 with multiple stepladder-like gas-fluid level in the bowel loops were noted.14 cases with completive obstruction and 13 cases with partial obstruction were found in UGI series.Malposition of the jejunum was seen in 8 cases among them.Abnormal location of cecum on barium enema was demonstrated in 30 cases,included 10 cases of microcolon.The etiology of duodenal obstruction included intestinal malrotation 34 cases,duodenal atresia 9 cases,duodenal stenosis 4 cases and anaular pancreas 5 cases.Conclusion Most cases of duodenal obstruction can be diagnosed with plain films combining with clinical materials.Differential diagnosis should be made by UGI series and barium enema.

Objective To detect Chlamydia trachomatis phage Vp1 gene in clinical swab specimens and anti-Vp1 antibodies in serum specimens.MethodsCervical and urethral swab as well as serum specimens were collected from attendees to the sexually transmitted disease(STD) clinic in the Tianjin Institute of STD,during March 2008 to March 2011.PCR was conducted to detect chlamydial phage Vp1 gene in swab samples,enzyme linked immunosorbent assay(ELISA) and Western blot to detect anti-Vp1 antibody in sera.The swab specimens positive for Vp1 gene were subjected to cell culture followed by the detection of Vp1 protein with an immunofluorescence-based method.ResultsTotally,36 out of 1542 swab specimens turned out to be positive for Vp1 gene,and 23 out of 453 serum specimens for anti-Vp1 antibody.No positive results were obtained in the Vp1 gene-positive swab specimens by cell culture and immunofluorescence-based assay.ConclusionThe Vp1 gene of Chlamydial trachomatis phage and anti-Vp1 antibody are successfully detected from clinical swab and serum specimens respectively.

Objective To discuss the imaging characteristics of chromophobic cell renal carcinoma (CCRC) and study the features on the contrast-enhanced ultrasound (CEUS). Methods The CEUS features of CCRC in 28 cases identified by pathology were reviewed. The blood supply and enhancement characteristic were observed and analyzed on time intensity curve parameters. Results The 28 cases of CCRC showed poor blood supply in contrast with the renal cortex. The CCRC presented with heterogeneity enhancement, part of the tumor took on a high wash-in and wash-out, and enhanced less intense than the surrounding renal cortex. The actinomorphous strong echo of the tumors might be revealed with CEUS in 15 cases (54%). The time intensity curve analysis demonstrated that the CCRCs' difference of peak intensity and area under the curve were lower than the renal cortex (P＜0.05), but arrival time, time-to-peak and slope of ascending curve were higher than the renal medulla (P＜0.05). Conclusion The actinomorphous enhancement and poor blood supply in the tumor of CEUS could provide diagnostic evidence for CRCC.

Objective To improve the knowledge of primary gout in children. Methods Clinical data of a 12-year-old girl with primary gout was collected. Analysis of UMOD gene, REN gene and HNF-1βgene was performed using PCR and di-rect sequencing. Results The girl was admitted for 1-month history of left hallux pain accompanied with elevations of serum uric acid concentration and serum creatinine concentration. Several examinations showed serum uric acid/creatinine ratio was greater than 2.5. The fractional excretion of uric acid was 3.4%-6.6%. The X-ray showed that the proximal phalanxes of halluces were erosion. The diagnosis of renal biopsy was ischemic renal injury and chronic tubulointerstitial nephropathy. Blood uric acid concentrations of parents were normal, and the family history of gout was negative. Two single nucleotide polymorphisms (c.264C>T heterozygous and c.866-71 G>A heterozygous) in UMOD gene, 1 single nucleotide polymorphism (c.373+44C>G heterozygous) in REN gene, and 2 single nucleotide polymorphisms (c.100-50-49ins TCTG heterozygous and c.781-22T>C homozygous) in HNF-1βgene were detected. No pathological mutation was detected in these 3 genes. Conclusions This child is highly suspected to have primary gout caused by familial juvenile hyperuricemic nephropathy.

Objective To get phagc's capsid Vp3 gene and protein of guinea pig inclusion conjunctivitis (GPIC) chlamydia. Methods The genome DNA was extracted from the φCPG1 phage.The full sequence of Vp3 gene was amplified by polymerase chain reaction (PCR) from the above genome DNA. The Vp3 gene was digested by restriction endonuclease and then inserted into prokaryotic plasmid vector pET30a (+). The recombinant plasmid was transformed into E. coil BL21, and was identified by restriction endonuclease, PCR and sequencing. The E. coil BL21 with expected recombinant plasmid was induced and the expressed recombinant Vp3 protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, then purified by agarose gel. Results The recombinant gene was sequenced and proved to be 447 bp which was consistent with the φCPG1 Vp3 gene sequence in GenBank. A 25 000 capsid protein was expressed and confirmed by SDS-PAGE and Western blot. The purified protein was obtained. Conclusion The capsid Vp3 protein of φCPG1 is successfully expressed and purified, which is helpful for the further study on its mechanism and clinical applications.

Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.

Objective To investigate the value of contrast enhanced ultrasound(CEUS) with microbubbles targeted to vascular endothelial growth factor receptor type 2 (VEGFR2) for imaging tumor angiogenesis in murine tumor models.Methods Established human renal cell carcinomas(RCC) subcutaneous xenograft tumor model in nude mice,microbubbles targeted to VEGFR2(MBV) was prepared,control microbubbles(MBC) and MBV were injected respectively in each mouse,the intensity of each microbubble was compared,the expressions of VEGFR2 in tumors were tested by immunohistochemistry.Results CEUS imaging showed the intensity of MBV and MBC was (6.50±1.43)dB,(2.59±0.99)dB,respectively.There was significantly higher intensity when using MBV compared with MBC (P<0.01).The time to wash out was longer in MBV contrast group compared with MBC group.Immunohistochemistry showed VEGFR2 was highly expressed in novel vessel walls.Conclusions Contrast microbubbles targeted to VEGFR2 can specially enhance the images of RCC and has tremendous significance in the assessment of angiogenesis.