Objective To detect the immunogenicity of VirB9,a protein of type Ⅳ secretion system of Brucella.Methods Full length VirB9 gene was cloned into plasmid pET32a and expressed in Escherichia (E.) coli BL21 (DE3).Expression of recombinant protein was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and the recombinant fusion protein was purified by affinity chromatography on Ni2+-conjugated chelateing sepharose.The purity of the purified protein was ascertained by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDSPAGE) and the concentration was measured by bicinchoninic acid (BCA) protein assay kit.Animal model was established by immunizing BAL B/c mice with live vaccine strain S19 of Brucella and the mice immunized with phosphate buffered saline (PBS) as control.The blood of immunized mice was acquired after 4 weeks.Antibody against VirB9 in S19 immunized mice was detected by Rose Bengal plate agglutination test and serum tube agglutination test; IgG antibody titers against VirB9 in immunized mice were determined by enzyme linked immunosorbent assay(ELISA).At the 35th day,the immunized mice and control mice were killed and spleens were collected.The splenocytes were harvested and stimulated with each of VirB9,concanvalin A(ConA) or medium in triplicate.Production of gamma interferon (IFN-γ) was determined by enzyme-linked immunospot assay (Elispot).Results The full length of VirB9 gene was cloned into pET32a.The recombinant VirB9 protein was expressed at 43 × 103 in relative molecular mass and the purity of the purified recombinant VirB9 protein was above 97％ in SDS-PAGE and the concentration was 1.6 g/L in BCA protein assay.The antibody of VirB9 was detected in all S19 immunized mice but not PBS immunized mice by Rose Bengal plate agglutination test.The antibody titer in all S19 immunized mice was ＞ 1 ∶ 800 or ＞ 1 ∶ 3200 by tube agglutination test and ELISA,respectively.Meanwhile,the protein stimulated stronger IFN-γresponse in immunized mice than that in the control mice(147 cells Vs 38 cells).Conclusion VirB9 can stimulate humoral and cellular immunity and it might be an appropriate target for developing subunit vaccine against Brucella.

[Objective] To detect the expression of NAD(P)H oxidase (Nox) and the content of ROS in different malignant gliomas,and explore the influence of Nox and intracellular ROS levels in the survival,proliferation,and malignant phenotype of gliomas.[Methods]Thirty human glioma specimens (10 with grade Ⅰ and I1,10 with grade II and 10 with grade IV),performed resection in our hospital from August 2007 to August 2010,were collected in our study;another 10 normal brain tissues were collected as controls.Real time PCR was used to detect the mRNA expressions of Nox(1-5);ROS level was detected by flow cytometry and Nox4 protein level was detected by immunofluorescence and Western blotting.[Results] The Noxl-5 mRNA and protein levels and ROS content were significantly different between each 2 groups (P＜0.05);Nox4 mRNA expression and ROS level significantly increased following the increment of malignancy degrees (controls＜low-grade gliomas＜grade III gliomas＜ grade IV gliomas,P＜0.05);the Nox protein expression was low in controls while that in gliomas was high,and the higher malignancy of the tumors,the higher expression levels of the tumors.[Conclusion] Nox expression and ROS content have significantly positive relations with the malignancy of the tumors,which indicates that Nox expression and ROS content might paly important roles in the occurrence of glioma and malignant proliferation.

To study the effects of short-term cryopreservation of cord blood hematopoietic cells in liquid nitrogen, the viability and function of cord blood hematopoietic cells were investigated by using each of 8 samples cryopreserved for six months, one and two years after thawing respectively. Nucleated cells (NC) were detected by blood cell analyzer. CD34+ cells were analyzed by flow cytometry, CFU-GM were cultured and detected in vitro, the survival rate was determined by trypan blue staining. The results showed that the differences of recovery rate of NC, CD34+, CFU-GM were nonsignificant at three different cryopreserved times. In conclusion, the short-term storage in liquid nitrogen showed a good effect on cord blood hematopoietic cell without any significant change of activities and number of the cryopreserved hematopoietic cells.
Cell Survival ; Cryopreservation ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Humans

Objective To study the clinical value of portable echocardiography system in diagnosis for acute paroxysmal dyspnea.Methods Clinical data of 81 patients with acute paroxysmal dyspnea recorded by a portable echocardiography apparatus at their bedside were retrospectively analyzed,and compared to those of 45 patients by conventional echocardiography.Results The 2D images in portable echocardiograph were similar to those of conventional echocardiograph.Diagnosis could be established in 74 (91.4%),corrected in six (7.4%) and not confirmed only in one (1.2%) of 81 patients with acute paroxysmal dyspnea by portable echocardiography system.And,portable echocardiography system could be used to diagnose pericardial effusion and to monitor perieardial puncturing and draining at bedside. Conclusions Portable echocardiography systems can provide rapid,accurate and valuable information on diagnosis and treatment for acute paroxysmal dyspnea,and make its clinical intervention accurate,scientific and effective,bringing echocardiography performed at bedside possible.

Adolescent ; Adult ; Athletic Performance ; psychology ; Back ; physiology ; Electromyography ; Exercise ; physiology ; Humans ; Imagery (Psychotherapy) ; Male ; Young Adult

OBJECTIVETo investigate the correlation of expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (Ki67) with sensitivity to neoadjuvant chemoradiation in rectal adenocarcinoma.

METHODSSamples of pretreatment biopsies and the resected specimens after neoadjuvant therapy in 32 patients with rectal adenocarcinoma were collected, and the expression of Ki67 and VEGF were detected by immunohistochemistry using specific antibodies. The correlation of Ki67 and VEGF expression with clinicopathological factors were analyzed.

RESULTSThe level of VEGF expression was significantly correlated with lymph node metastasis (P = 0.033), depth of tumor invasion (P = 0.007) and TNM stage (P = 0.016), but not with histological type, tumor size, age and gender of the patients (P > 0.05). However, VEGF expression was found to be negatively and significantly correlated with the sensitivity to neoadjuvant chemoradiation (P = 0.016), and a transient increase of VEGF expression was detected in the resected specimens after neoadjuvant therapy (P = 0.035). Ki67 labeling index (Ki67-LI) was found to be significantly correlated with lymph node metastasis (P = 0.028), but not with tumor size, age and gender of the patients (P > 0.05). It was also found that tumors with lower Ki67-LI expression were more sensitive to neoadjuvant therapy than that with higher expression of Ki67-LI (P = 0.032). In contrast with VEGF, the Ki67 expression level decreased after neoadjuvant therapy, but no statistical significance was found between pretreatment and posttreatment specimens (P > 0.05).

CONCLUSIONThe preliminary results of this study demonstrate that the expression of VEGF and Ki67 in pretreatment biopsy of rectal adenocarcinoma may be used as a biomarker to predict tumor response to neoadjuvant chemoradiation.

Adenocarcinoma ; metabolism ; pathology ; surgery ; therapy ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Female ; Humans ; Ki-67 Antigen ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Invasiveness ; Neoplasm Staging ; Radiotherapy, Conformal ; Rectal Neoplasms ; metabolism ; pathology ; surgery ; therapy ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVETo explore the anatomical structures, and depth and direction of needling at Jingming (BL 1), so as to provide anatomical basis for its clinical application.

METHODSForty-eight adult orbital specimens were observed by dissection.

RESULTSWhen a needle was vertically inserted into Jingming (BL 1), the needle tip will past through the skin, subcutaneous tissue, medial palpebral ligament, medialis rectus and orbital adipose body. Above the body of the needle, there are ophthalmic artery, anterior ethmoidal artery and nasociliary nerve. The average distance between the skin at the punctured point and the anterior ethmoidal artery is (18.25 +/- 4.45) mm, with an angle of (12.5 +/- 5.5) degrees, and the average distance between the skin at the punctured point and the optic nerve tunnel frontal point is (43.37 +/- 7.84) mm.

CONCLUSIONTo avoid bleeding caused by injuring the anterior ethmoidal artery, acupuncture at Jingming (BL 1) should avoid deeply inserting needled back-upwards and upwards, and the needling depth should not exceed 30.36 mm to avoid injury of the optic nerve tunnel frontal point.

Acupuncture Points ; Female ; Humans ; Male ; Orbit ; anatomy & histology

OBJECTIVETo observe the distribution of the blood vessels in the integument tissue of the channel area of legs.

METHODSThe integument tissue of the lower limbs in the 12 cadavers were dissected with macro-and micro-dissection, radiographical technique of systemic artery and technique of image pattern analysis to observe and analyze the origins, branches and anastomoses in the integument tissues along the channels of legs.

RESULTSThe distributional density of the blood vessels in the integument tissues of legs along the channel area of the three-yin meridians of the foot, the Gallbladder Meridian, and the Urinary Bladder Meridian was higher than that in the other parts. They formed an obvious nutrient vascular chain on the arteriogram. The distributional density in the channel area of the Stomach Meridian was not obviously increased and the obvious nutrient vascular chain could not be seen.

CONCLUSIONAn obvious nutrient vascular chain is formed in the integument tissue along the channel area of legs except the Stomach Meridian.

Blood Vessels ; anatomy & histology ; Humans ; Leg ; blood supply ; Meridians

Objective To observe the efficacy and safety of applying sirolimus-eluting stents in emergency percutaneous coronary intervention (PCI) for the patients with acute myocardial infarction (AMI).Methods In total,220 patients with AMI were enrolled in this study at Shanghai Tongji Hospital, divided into two groups,one with bare-metal stent and the other with sirolimus-eluting stent.Cardiovascular fatality,major adverse cardiac events (MACE) and target vessel revascularization (TVR) were observed one and six months after PCI in the two groups.Results There was no significant difference in overall fatality and MACE in the 1~(st) or 6~(th) months after PCI between the two groups.Three cardiogenic deaths occurred in bare-metal stent group with a fatality of 2.8 percent,and five deaths in sirolimus-eluting stent group with a fatality of 4.5 percent in six months after PCI.However,rate of restenosis in those with sirolimus-eluting stents was significantly lower than that of bare-metal stents (6.0 percent vs 16.1 percent,P

OBJECTIVEPituitary prolactinoma with severe erectile dysfunction (ED) as the initial symptom is often misdiagnosed. This article explores the diagnosis and treatment of severe ED caused by pituitary prolactinoma.

METHODSWe retrospectively analyzed the diagnosis and treatment of 4 cases of pituitary prolactinoma with severe ED (IIEF-5 score 5 - 7) as the initial clinical symptom confirmed by MRI.

RESULTSThe 4 cases of pituitary prolactinoma-induced severe ED, with serum prolactin 10 times above the maximum normal level, were misdiagnosed for 2 years. All failed to respond to the PDE5 inhibitor therapy, and then 3 of them underwent transnasal hypophysectomy. Twenty-four months of follow-up found the level of prolactin restored to normal in 1 case (IIEF-5 = 19), and reduced to 600 and 768 IU/L respectively (IIEF-5 = 15) in the other 2. Then administration of the PDE5 inhibitor was followed, which produced satisfactory efficacy. One case was treated with oral bromocriptine, which restored the prolactin level to normal at 12 months (IIEF-5 > 21).

CONCLUSIONProlactin detection and brain MRI can help to confirm pituitary prolactinoma with severe ED at the onset. As for its treatment, in case of an extremely high level of prolactin, simple administration of the PDE5 inhibitor is ineffective. When the prolactin level is reduced after surgery or medication, the symptom of ED can be improved and, in case of no obvious relief, administration of the PDE5 inhibitor can be followed, which may achieve satisfactory results.

Adult ; Erectile Dysfunction ; diagnosis ; etiology ; Humans ; Male ; Middle Aged ; Phosphodiesterase 5 Inhibitors ; therapeutic use ; Pituitary Neoplasms ; complications ; diagnosis ; drug therapy ; Prolactinoma ; complications ; diagnosis ; drug therapy ; Retrospective Studies