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 investigate the effects of television-watching and computer-using on sleep/wake patterns, sleep duration and sleep problems of school-aged children in Shanghai. Methods A total of 4 108 school-aged children from 10 primary schools of Shanghai were enrolled by multi-stage cluster sampling and surveyed by questionnaires. The information of television-watching and computer-using, family and personal condition was investigated by self-prepared questionnaire, and the Chinese version of Children's Sleep Habits Questionnaire was employed to survey the sleep behaviors of children. The effects of television-watching and computer-using on sleep/wake patterns, sleep duration and sleep problems were analyzed by multiple linear regression analysis and Logistic regression analysis. Results The percentage of children who watched television≥2 h per day was 4.1% during weekdays, and that came to 49.2% during weekends. In terms of frequency of computer-using, most children reported "rarely" (88.2%, 0-1 time/week), followed by "often" (11.0%, 2-4 times/ week) and "usually" (0.8%, 5-7 times/week). With the age increase, the percentages of children who watched television≥2 h per day and those who "often" used computer gradually increased. It was revealed by multiple linear regression analysis and Logistic regression analysis that television-watching and computer-using were not only positively correlated with later bedtime, later wake time and shorter sleep duration but also significantly associated with sleep problems such as bedtime resistance, sleep onset delay, sleep duration disorder, sleep anxiety and parasomnia. Conclusion Television-watching and computer-using exert influences on sleep behaviors of sleep/wake patterns, sleep duration and sleep problems. Concerns about the potential negative effects of television-watching and computer-using on sleep behaviors may help to promote healthy sleep patterns and improve sleep quality.

AIMTo investigate the effect of hyperthermia on apoptosis of cultivated striatum neurons in the rat.

METHODSAfter 30 min hyperthermia in 43 degrees, the Ca2+ concentration, the mitochondria membrane potential of the neuron were detected by Laser Scanning Confocal Microscopy (LSCM). The apoptosis of striatum neurons was detected by TUNEL staining.

RESULTSHeat stress at 43 degrees C for 40 min caused an increase in the Ca2+ concentration of striatum neurons and a decrease in the mitochondria membrane potential of the striatum neurons.

CONCLUSIONThe striatum shows more apoptosis neurons after heat stress.

Animals ; Animals, Newborn ; Apoptosis ; Cells, Cultured ; Corpus Striatum ; cytology ; Heat-Shock Response ; Hot Temperature ; adverse effects ; Membrane Potential, Mitochondrial ; Neurons ; cytology ; pathology ; Rats ; Rats, Wistar

Objective To analyze the diagnosis and treatment for complications of renal caliceal diverticulum with calculi or infection. Methods A retrospective investigation was performed on 29 cases with renal caliceal diverticulum. The 29 cases included 11 males and 18 females aged 18 to 61 years. Among the study group, 3 cases were simple renal caliceal diverticulum, 12 cases were diagnosed as diverticular calculi and 14 cases presented recurrent urinary tract infections including 3 cases with urinary fistula after unroofing and decompression as renal simple cyst from another hospital. Ten cases underwent an open operation that unroofed and decompressed the cyst, and sutured the diverticular neck. Eight cases underwent laparoscopic operation similar to the open operation, including lithotomy in caliceal diverticulum in 2 cases. Eleven cases diagnosed with caliceal diverticular calculi were taken one-stage percutaneous nephrolithotomy including dilating the diverticular neck, remaining the nephrostomy catheter and Double-J ureteral stents, and 1 case was transferred to open operation.Results The open and laparoscopic operations were performed successfully. One case was cured by Double-J ureteral stenting after postoperative urinary leakage. One case was transferred to open operation for the failure of percutaneous puncturation. X-ray examination revealed that there were no remaining stones after the operation. All the patients were followed up for 6 to 24 months without calculi and infection recurrence. Conclusions Stones and infection are common that complications of renal caliceal diverticulum. Percutaneous nephrolithotomy, laparoscopy and other operations were effective and feasible treatment options for cases with complications of renal caliceal diverticulum. Exact diagnosis was very important for treatment of renal caliceal diverticulum before operation.

Objective To compare the diagnostic value of myocardial perfusion imaging (MPI) and 64 multi-slice spiral CT (64-MSCT) for coronary artery disease (CAD). Methods Fifty-two patients with suspected or known CAD were included in the study. Each patient underwent both stress and rest MPI,MSCT as well as conventional coronary angiography (CAG) within 1 month. The stress and rest MPI were scored by a 5-grade criteria (0 ～ 4) based on 17 coronary artery segments. The difference between summed stress and rest scores ＞ 1 was defined as myocardial ischemia. Stenosis in one main vessel or one main branch of the main vessel ≥50% was defined as myocardial ischemia by MSCT. CAG was used as the reference for comparison. Statistical analysis was performed using SPSS 13. 0 software. Kappa value was used to test the accordance of MPI and MSCT results. X2 test was used to evaluate the difference between MPI and MSCT results. Results The patient-based sensitivity, specificity, positive and negative predictive values and accuracy of MPI and MSCT for the diagnosis of CAD were 86.7% (26/30), 77.3% ( 17/22),83.9% (26/31), 81.0% ( 17/21), 82.7% (43/52) and 83.3% ( 25/30), 86.4% ( 19/22), 89.3%( 25/28), 79.2% ( 19/24), 84.6% (44/52), respectively. The vessel-based sensitivity, specificity, positive and negative predictive values and accuracy of MPI and MSCT were 74.5% (38/51), 81.0% (85/105 ), 65.5% (38/58), 86.7% ( 85/98), 78.8% ( 123/156 ) and 90.2% (46/51 ), 88.6% ( 93/105 ),79.3 % (46/58), 94.9% (93/98), 89.1% ( 139/156), respectively. There was no statistically significant difference between MPI and MSCT for either patient or lesion-based diagnosis (X2 =0.44, 0.21, both P ＞0.05 ). 96.0% (24/25) patients with both abnormal MPI and MSCT positive were valified by CAG while 83.3% (15/18) patients with both MPI and MSCT negative were excluded by CAG. Conclusions Both MPI and MSCT are reliable diagnostic modalities for CAD. They also provide complementary diagnostic value to each other.

AIM To investigate the effect and mechanism of Qigui Yishen Decoction (QGYS,Astragali Radix,Angelicae sinensis Radix,Chuanxiong Rhizoma,Achyranthis bidentatae Radix) on regulating the expression of miR-141 in unilateral ureteral obstruction (UUO) mice with renal fibrosis.METHODS Thirty Balb/c male mice randomly divided into sham-operated group (n =6),UUO group (n =6),Lotensin (50 g/kg) group (n =6),QGYS high dose (50 g/kg) group (n =6),and QGYS low dose (10 g/kg) group (n =6) were conducted UUO surgery to promote kidney fibrosis except the six mice in the sham operation group.After a successive 10-day medication of QGYS and Lotensin to mice by oral gavage on daily basis,all mice were killed to procure renal tissue to observe its morphology and pathology changes by HE staining.The expressions of TGF-β1,ColⅣ,and MMP-9 were analyzed by immunohistochemical method,and the expressions of miR141,TGF-β1 were measured by real-time PCR.RESULTS The obviously pathological injuries including renal interstitial fibrosis were identified by HE staining among the groups intervened with UUO,but the variance in the extent due to different administrations of QGYS and Lotensin was noticed as well (P ＜ 0.05).As compared to the UUO group,high and low dose QGYS groups and Lotensin group achieved an up-regulated expression of TGF-β1 and ColⅣ,and a down-regulated expression of MMP-9 by immunohistochemistry (P ＜ 0.05),and significantly increased Mrna expression of miR-141,and decreased Mrna expression of TGF-β1 by real-time PCR (P ＜ 0.05).CONCLUSION In UUO mouse models,QGYS gives influence to TGF-β1and MMP-9 through inducing miR-141 expression change to decrease abnormal accumulation of ECM,and thus inhibits the progression of renal fibrosis.

The present study is to assess the prophylactic effect of quinacrine (QA) , an anti-malarial drug, against heatstroke in rats. Conscious rats were orally given equal volume normal saline or QA (dissolved in normal saline and final dosage for rats was 4.5, 9.0 and 18 mg x kg(-1)). An hour later rats were put into a warm water circulated hot chamber (41.0 +/- 0.5) degrees C. Rectal temperature (core temperature, T(co)) of rats in hot chamber was continuously monitored by a thermocouple. T(co) and survival time of rats showed that QA pre-treatment postponed the hyperthermia, and increased the survival time of rats in hot chamber. Primary striatum neurons' culture from new born rats was maintained with D-MEM and 10% FBS. After immuno-cytochemistry identification with antibodies against neural specific proteins, culture received 20 micromol x L(-1) QA only for 1 h and followed by 43.0 degrees C heat treatment for another hour, or 20 micromol x L(-1) QA for 1 h followed by 43.0 degrees C heat treatment for another hour. Control culture received heat treatment only. Cultures were labeled with the fluorescent indicator DPH and the relative membrane fluidity of neurons was measured with the help of fluorescent polarized spectrophotometer. [3H] Arachidonic acid (AA) labeled membrane of E. Coli cells was used as substrate to determine cPLA2 activity of neurons. Gas chromatography and mass spectrum were also employed to detect on the level of fatty acids level in rat striatum neurons. Results from cells indicated that inhibition of cPLA2, reduction the release of active fatty acids such as AA, and possibly, stabilization of the cell membrane which was disturbed by hot treatment, may contribute to the mechanism underlying heat protection and heatstroke preventive effects of quinacrine.
Animals ; Cells, Cultured ; Corpus Striatum ; drug effects ; pathology ; Fatty Acids ; metabolism ; Heat Stroke ; metabolism ; physiopathology ; prevention & control ; Hot Temperature ; adverse effects ; Male ; Membrane Fluidity ; drug effects ; Neurons ; enzymology ; metabolism ; physiology ; Phospholipases A2 ; metabolism ; Quinacrine ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of different anesthetic and analgesic protocols on the cellular immune function and stress hormone in patients undergoing lobectomy for esophagus cancer.

METHODSSixty ASA I or II patients undergoing lobectomy for esophagus cancer were randomly divided into two groups to receive postoperative general anesthesia and intravenous analgesia (group A, n=30) or intraoperative general anesthesia combined with thoracic epidural anesthesia with postoperative epidural analgesia (group B, n=30). The cervical venous blood samples were obtained from the patients at 30 min before anesthesia induction (T(0)), 2 h after skin incision (T(1)), and at 4 h (T(2)), 24 h (T(3)) and 48 h (T(4)) after the end of operation. The T-lymphocyte subsets (CD4(+) and CD8(+)) were analyzed by flow cytometry, serum concentrations of sIL-2R and IL-2 determined by ELISA, and the levels of growth hormone (GR), prolactin (PRL), IL-8 and cortisol (Cor) measured by radioimmunoassay. Visual analogue scale (VAS) was used for assessment of the postoperative analgesic effects.

RESULTSThe VAS scores were significantly lower in group B than in group A at T(2) and T(3) (P<0.05). The percentage of CD4(+) cells and the CD4(+)/CD8(+) ratio in the two groups began to decrease significantly at T(1) (P<0.05), reducing to the lowest level at T(2) in group B and at T(3) in group A. From T(1) to T(4), the percentage of CD4(+) in group B remained significantly higher than those in group A (P<0.05), and from T(3) to T(4), the CD4(+)/CD8(+) ratio in group B were significantly higher than those in group A (P<0.05). The IL-2 level in the two groups began to decrease significantly at T(1) (P<0.05), reaching the lowest level at T(2) in group A and at T(3) in group A. IL-2 level was significantly higher in group B than in group A from T(3) to T(4) (P<0.05). sIL-2R level in group A began to increase at T(1) and peaked at T(3), showing significant differences from the T(0) level, but the level showed no significant variations in group B compared with the T(0) level. From T(2) to T(4), sIL-2R level was significantly higher in group A than in group B (P<0.05). The levels of GH, PRL and Cor increased significantly, while IL-8 decreased in the two groups from T(1) to T(4) (P<0.05), but remained stable in group B.

CONCLUSIONGeneral anesthesia combined with thoracic epidural anesthesia may reduce the perioperative stress reaction and adverse effect on cellular immune function in patients undergoing lobectomy for esophagus cancer.

Adult ; Aged ; Analgesia, Epidural ; methods ; Anesthesia, Epidural ; methods ; Anesthesia, Intravenous ; methods ; Esophageal Neoplasms ; immunology ; surgery ; Female ; Humans ; Immunity, Cellular ; drug effects ; Male ; Middle Aged ; Pain, Postoperative ; drug therapy ; Postoperative Period ; Stress, Physiological ; drug effects ; T-Lymphocytes ; immunology

AIMTo study the effect of chronic hypoxic hypercapnia on expression of COX-2 mRNA in pulmonary arterioles.

METHODSSD rats were randomly divided into two groups: control group and hypoxic hypercapnic group. COX-2 mRNA was observed in pulmonary arterioles by the technique of in situ hybridization.

RESULTSmPAP, weight ratio of right ventricle (RV) to left ventricle plus septum (LV + S) and COX-2 mRNA in pulmonary arterioles were much higher in rats of hypoxic hypercapnic group than those of control group. Light microscopy showed that vessel smooth muscle cell hypertrophy and vessel cavity straightness were found in hypoxic hypercapnic group.

CONCLUSIONChanges of expressions of COX-2 mRNA may regulate hypoxic hypercapnic pulmonary hypertension.

Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Hypercapnia ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the prevalence of chronic prostatitis in men with premature ejaculation.

METHODSThe segmented urine specimens before and after prostatic massage and the expressed prostatic secretion specimens from 106 patients with premature ejaculation and 38 controls were evaluated by microscopic and/or bacteriological studies. The prevalence of premature ejaculation was also investigated in 120 patients with chronic prostatitis.

RESULTSProstatic inflammation was found in 46.2% and chronic bacterial prostatitis in 34.7% of the subjects with premature ejaculation, respectively. Compared with the controls, the findings were statistically significant (P < 0.05). The prevalence of premature ejaculation in the patients with chronic prostatitis was 47.5% (57/120).

CONCLUSIONSChronic prostatic inflammation may play a role in the pathogenesis of some cases of premature ejaculation and it is important to give a careful examination of the prostate before initiating any therapy for premature ejaculation.

Adult ; Chronic Disease ; Ejaculation ; Humans ; Male ; Middle Aged ; Prevalence ; Prostate ; diagnostic imaging ; Prostatitis ; complications ; epidemiology ; Sexual Dysfunction, Physiological ; etiology ; Sexual Dysfunctions, Psychological ; etiology ; Ultrasonography