[Effects of 4'-O-methylochnaflavone on endothelial dysfunction induced by palmitic acid in rat cavernous endothelial cells].

OBJECTIVE: To investigate the effect of biflavonoid 4'-O-methylochnaflavone (MF) on palmitic acid-induced endothelial dysfunction in rat cavernous endothelial cells (RCECs). METHODS: The isolated RCECs were commercially available and randomly divided into four groups: normal+BSA group (NC group), palmitic acid (PA) group, MF group, and icariside Ⅱ (ICA Ⅱ) group. The protein expression levels of protein kinase B (PKB/AKT) and endothelial nitric oxide synthase (eNOS) in each group were evaluated via Western blotting. The differences in the intracellular nitric oxide of RCECs treated by MF or ICA Ⅱ were detected by DAF-FM DA that served as a nitric oxide fluorescent probe. Effects of MF and ICA Ⅱ on cell proliferation of PA-stimulated RCECs were determined via CCK-8 assay. RESULTS: The content of nitric oxide in RCECs was significantly increased after the treatment of MF and ICA Ⅱ in comparison with the NC group (P < 0.05). Moreover, compared with ICA Ⅱ group, MF demonstrated a more obvious effect in promoting nitric oxide production (P < 0.05). Compared with the NC group, the expression levels of eNOS and AKT in the PA group were significantly decreased, indicating that a model for simulating the high-fat environment in vitro was successfully constructed (P < 0.05). Meanwhile, the intervention of MF and ICA Ⅱ could effectively increase the expression of eNOS and AKT, suggesting that MF and ICA Ⅱ could promote the recovery of endothelial dysfunction caused by high levels of free fatty acids (P < 0.05). The results of CCK-8 assays showed that PA could significantly reduce the proli-feration ability of RCECs (P < 0.05). Furthermore, the decreased cell viability induced by PA was significantly elevated by treatment with ICA Ⅱ and MF (P < 0.05). CONCLUSION: In RCECs, MF and ICA Ⅱ could effectively increase the content of nitric oxide. The down-regulation of the expression of proteins associated with the AKT/eNOS pathway after PA treatment revealed that this pathway was involved in the development of endothelial dysfunction, which could be effectively reversed by MF and ICA Ⅱ. In addition, the cell proliferation ability was significantly decreased following PA treatment, but MF and ICA Ⅱ could restore the above changes. Overall, biflavonoid MF has an obvious repairing effect on PA-stimulated endothelial dysfunction.

[Clinicopathological characteristics of type 2 diabetes mellitus complicated with colorectal cancer].

Objective: To analyze the clinicopathological features of type 2 diabetes mellitus complicated with colorectal cancer (DCRC). Methods: A case-control study was conducted. Inclusion criteria: (1) hospitalized patients receiving fibrocolonoscopy; (2) adenocarcinoma and mucinous adenocarcinoma diagnosed by pathology; (3) with preoperative cTNM clinical staging; (4) colorectal cancer patients undergoing surgical treatment; (5) with postoperative pTNM staging; (6) no smoking or drinking habits. Exclusion criteria: (1) familial adenomatous polyposis (FAP); (2) Lynch syndrome; (3) carcinoma of anal canal and perianal carcinoma; (4) multiple primary cancer; (5) with serious cardiocerebrovascular diseases or multiple organ failure. Clinicopathlogical data of 32 DCRC patients who were diagnosed and treated in Peking University Shougang Hospital from December 2017 to December 2018 were retrospectively collected and analyzed. Forty nondiabetic colorectal cancer (CRC) patients during the same period were selected as control group according to the sex ratio and the age difference less than 5 years. Student's t test and χ(2) test were used to compare the difference between the two groups in baseline clinicopathological data, clinical test results, tumor markers and infiltration status of T cells in tumor immune microenvironment. Results: Among 32 DCRC patients, 24 were males and 8 were females with a mean age of (63.0±1.7) years; among 40 CRC patients, 30 were males and 10 were females with a mean age of (60.5±1.6) years. The duration of diabetes mellitus in DCRC patients (from the diagnosis of diabetes mellitus to the diagnosis of colorectal cancer) was (9.2±1.3) years. The body mass index (BMI) of DCRC group was significantly higher than that of CRC group [(24.8±0.6) kg/m(2) vs. (23.2±0.4) kg/m(2), t=2.372, P=0.020]. There were no significant differences in other baseline data (sex, age, primary site of tumor, R0 resection rate, pathological stage, pathological type, differentiation degree of tumor, preoperative intestinal obstruction) between the two groups (all P>0.05). Serum triglyceride level in DCRC group was higher than that in CRC group [(2.1±0.2) mmol/L vs. (1.5±0.1) mmol/L, t=3.085, P=0.003], while hemoglobin [(120.3±5.2) g/L vs. (132.7±2.8) g/L, t=-2.224, P=0.029], anti- thrombin III [(94.2±3.7)% vs. (103.5±2.4)%, t=-2.197, P=0.031], and red blood cell count [(4.2±0.1)×10(12)/L vs. (4.5±0.1)×10(12)L, t=-2.055, P=0.044] were all lower than those in CRC group. The preoperative carcinoembryonic antigen (CEA) level in DCRC group was higher than that in CRC group [(50.3±21.8) µg/L vs. (5.6±1.0) µg/L, t=2.339, P=0.022]. There were no significant differences in preoperative levels of other four tumor molecular markers (CA199, CA242, CA724 and CA125) between the two groups (all P>0.05). The expression of Foxp3 [specific markers of CD4+, CD25+ regulatory T cells (Treg)] in DCRC group was higher than that in CRC group [(82.7±6.2) cell/HPF vs. (62.6±4.9) cell/HPF, t=2.586, P=0.012]. There were no significant differences in the infiltration of CD4, CD8, PD-1 and PD-L1 positive cells between two groups (all P>0.05). Conclusions: The average diabetic history of DCRC patients is nearly 10 years. They have higher BMI and serum CEA level, and more Treg cell infiltration in the tumor. Close attention should be paid to these patients in clinical practice.

[The outcomes comparison between autologous incus and titanium PORP in Austin A and C type ossicle defect cases].

Objective: A series of otitis media patients with Austin A and C type ossicles defects received ossicular chain reconstruction surgery using either autologous incus or titanium partial ossicular replacement prosthesis (PORP).The hearing outcomes and exclusion rate of prosthesis were compared between these two groups. Method: One hundred and seventy-six patients with chronic otitis media underwent ossiculoplasty in our hospital during 2016-2017. These patients were divided into 2 groups: group A consisted of those receiving autologous incus replacement (n=85), and group B consisted of those receiving titanium PORP (n=91). All the patients received CWD mastoidectomy and ossiculoplasty in one stage and were followedup in postoperative 3, 6 and 12 months. Result: The postoperative hearing improvement rates in group A and B were 85.88% and 92.31%, respectively, and there was no statistical difference. No statistical difference of the postoperative hearing improvement rates was observed between Austin A and C type in 2 groups. Three cases (3.29%) had ossicle exclusion in B group, and 0 case in A group, which was not statistical different (P>0.05). Conclusion: Patients receiving ossicular chain reconstruction surgery using autologous incus show similar hearing outcomes as those using titanium PORP. The risk of prosthesis exclusion is lower for autologous incus than titanium PORP. The residual handle of malleus has no obvious influence on the postoperative hearing improvement.

[The differences of myringoplasty performed in active and inactive nflammatory stage].

Objective:Through comparing of simple underlay myringoplasty performed in active and inactive inflammatory stage of the chronic otitis media, to explore how the tympanic cavity inflammation affect the outcome of the myringoplasty.Method：Forty-six ears were enrolled in active inflammation group (group A,n=46), and 52 ears were enrolled in inactive inflammation group (group B,n=52). All the patients had received the simple underlay technique myringoplasty. The following-up with otoscope and auditory test had been done in post-operative 12 months.Result:There was no statistic difference of gender and age between two groups. The graft taken-up rate of the group A was 87,0%, that of group B was 90,4%, and there was no statistic difference between two groups(P>0.05).There was no statistic difference of auditory threshold in pre-operative period(P>0.05) and the ABG closure in post-operative period(P>0.05)between two groups. The re-perforation rate of the tympanic membrane also has no difference between two groups(P>0.05).Conclusion：There is no difference of the underlay technique myringoplasty between the active inflammatory stage and inactive inflammatory stage. The active inflammatory condition of the tympanic cavity is not the contraindication of the myringoplasty.

Twist2 and CD24 expression alters renal microenvironment in obesity associated kidney cancer.

OBJECTIVE: Obesity emerged as a major public health problem worldwide, and prolonged condition with increased BMI causes various metabolic disorders include the development of kidney cancer. The metabolic changes alter the renal microenvironment and thereby promoting tumor. Hence, detailed studies of genes that regulate these this changes are keen to understand. MATERIALS AND METHODS: Initially, we successfully initiate kidney tumor using prolonged intake of a high-fat diet in Wistar rats, which are confirmed with pathological changes observed through histological sectioning. The expression of Twist2 and CD24 was assessed using Immunohistology and Western Blotting in a different time interval of kidney cancer. RESULTS: The rats fed with high-fat diet for 8 months shows 1.5 times increased in body mass whereas rats fed with high-fat diet for 16 months shows triple the size when compared with controls. Histological sectioning confirms the development of lesions and proteinaceous casts in 8 months high-fat fed rats, whereas we observed the high proliferative mass of cells in 16 months high-fat fed rats. Interestingly, we also observed elevated expression of Twist2 in initial stages of kidney cancer, which are down-regulated in the latter stages of kidney cancer. The experiments with CD24 shows the gradual increase of the expression of CD24 as a tumor develops to the next level. CONCLUSIONS: The correlation between Twist2 and CD24 expression conclude that Twist2 overexpression in initial stage augments CD24 to express more in the latter stage of kidney cancer. Reversely, the overexpression of CD24 and down-regulation of Twist2 in later stages of kidney cancer suggest the CD24 expression is dependent on Twist2 expression level.

[Epidemiological investigation of allergic rhinitis in Ningxia].

Objective: To investigate the epidemiological characteristics of allergic rhinitis (AR) in Ningxia and to analyze its related factors. Methods: From March to September of 2013, a multi-stage and cluster sampling method was used to investigate the diagnosis and treatment of AR in Ningxia Area (3 years and above). Guidelines for diagnosis and treatment of allergic rhinitis (2009, Wuyishan) was used as the basis for the diagnosis of adult AR, while Guidelines for diagnosis and treatment of pediatric allergic rhinitis (2010, Chongqing) was used as the basis for children. SPSS 16.0 software was used to complete the statistical analysis. Results: The total number of questionnaires was 6 000, and the number of effective questionnaire was 5 236, the recovery rate was 87.27%. With 684 cases diagnosed of AR, the prevalence of AR in Ningxia was 13.06% (684/5 236), including 13.40% (325/2 425) of males, 12.77% (359/2 811) of females. The difference was not statistically significant (χ(2)=0.456, P>0.05). There was significant difference in the prevalence between Hui and Han [14.35% (452/3 150) vs 11.12% (232/2 086), χ(2)=11.51, P<0.05]. According to ARIA criteria, persistent AR was 27.63% (189/684), intermittent AR was 72.37% (495/684). The month with highest incidence of AR in Ningxia Area was September, accounting for 71.78% (491/684). The prevalence of urban population was 14.54%, with the prevalence of rural population was 11.90%, and the difference was significant between urban and rural residents (χ(2)=7.90, P<0.05). The age group with highest prevalence rate was 21~30 years old. The main inhalation allergens were mugwort (68.42%), weeds (58.48%) and ragweed (55.56%). The main dietary allergens were wheat flour (14.33%), peanut (13.74%) and walnut kernel (11.99%). The most common complication was allergic conjunctivitis [82.02% (561/684)]. Conclusion: The epidemiology of AR in Ningxia Area is preliminarily understood, which will provide the epidemiological evidence for the prevention and treatment of AR and the formulation of public health policy.

Association between folate status and cervical intraepithelial neoplasia.

BACKGROUND/OBJECTIVES: To investigate the effect of folate status on cervical intraepithelial neoplasia (CIN) progression and its relationship with high-risk human papillomavirus (hrHPV). SUBJECTS/METHODS: We evaluated 20 000 sexually active women aged <65 years in Yangqu County by using a questionnaire; the subjects were also screened using the ThinPrep cytologic test (TCT). Patients with abnormal TCT results (other than glandular cell abnormalities) who were willing to provide informed consent were further diagnosed using colposcopy and histopathological examination. We investigated 247 cases of low-grade cervical squamous intraepithelial lesions (LSIL), 125 cases of high-grade cervical squamous intraepithelial lesions (HSIL) and 877 controls. A 24-item food frequency questionnaire was filled out by the investigator to estimate the consumption of dietary folate. Positivity for hrHPV from residual exfoliated cervical cells was tested; serum folate was also measured. RESULTS: The hrHPV infection rate in HSIL patients (77.6%) was higher than that in LSIL (33.2%) and control (32.0%) patients. Dietary folate intakes in controls, LSIL and HSIL were 306.9±176.6, 321.8±168.0 and 314.7±193.8 µg/kcal, respectively. The levels of serum folate in controls, LSIL and HSIL were 18.2±7.9, 15.9±7.1 and 14.3±7.5 nmol/l, respectively. Increased CIN correlated with higher rates of hrHPV infection and lower levels of serum folate. CONCLUSIONS: Low levels of serum folate may increase the risk of CIN progression. Furthermore, potential synergy may exist between low serum folate levels and hrHPV infection to promote CIN development.

[Curative observation on allergic rhinitis treated by intranasal corticosteroids combined with nasal irrigation].

Objective: To observe the effect of normal saline with intranasal corticosteroids in the treatment of allergic rhinitis(AR).Method:Two hundred and seventy-three outpatients with severe persistent allergic rhinitis were participated in this study.All patients were randomly divided into three groups, and were treated for 12 weeks using three different therapies, respectively. Symptom assessment of each patient were performed before treatment, 4 weeks, 8 weeks, and 12 weeks after treatment, respectively. The content of symptom assessment includes visual analogue scale, rhinoconjunctivitis quality of life questionnaire, symptoms scores and endoscopy examination scores.Result:Efficacy of intranasal corticosteroids combined with normal saline nasal irrigation in the treatment of AR is better than that of intranasal corticosteroids only, and is almost equivalent to intranasal corticosteroids combined with antihistamines.Conclusion:Long-term application of intranasal corticosteroids in combination with normal saline irrigation can effectively control and mitigate symptoms of severe persistent AR patients, and improve the patient's quality of life. This combination therapy is a better choice for the patient who is not satisfied with the single use of intranasal corticosteroids.

The efficacy and safety of zibotentan in the treatment of castration-resistant prostate cancer: a meta-analysis.

OBJECTIVE: Recently, novel endothelins like zibotentan and atrasentan and other novel taxanes have been introduced to treat prostate cancer. This study reviews zibotentan in the treatment of castration-resistant prostate cancer (CRPC) and derives a more precise estimate of their effect of treatment. MATERIALS AND METHODS: Two reviewers searched and extracted data of the published trials and review articles on zibotentan for prostate cancer using the Medline, Embase and Cochrane Controlled Trials Register database. We used hazard ratios (HRs) to assess the effects on overall survival (OS), progression-free survival (PFS), or time to PSA progression (TTP), and relative risk (RR) for the different types of toxicity. Four randomized controlled trials were identified. RESULTS: The pooled HR showed that zibotentan did not improve OS and PFS (HR = 0.92, 95%CI = 0.82-1.03, p = 0.161, HR = 0.98, 95% CI = 0.89-1.08, p = 0.714). Zibotentan had modest benefits on TTP (HR = 0.94, 95% CI = 0.91-0.97, p = 0.001). In addition, zibotentan led to more peripheral edema, anemia, cardiac failure and pneumonia. CONCLUSIONS: Our study concludes that zibotentan is not an attractive option for CRPC patients. However, additional studies on other novel therapies are needed to improve patient outcomes.

A pilot study of endoscopic spray cryotherapy by pressurized carbon dioxide gas for Barrett's esophagus.

BACKGROUND AND STUDY AIMS: Endoscopic spray cryoablation is a novel approach for the treatment of Barrett's esophagus. However, few studies have reported its efficacy, especially with the use of carbon dioxide (CO (2)). The aim of the current study was to evaluate the short term efficacy and complications using CO (2) in endoscopic cryoablation of Barrett's esophagus. METHODS: Patients diagnosed with Barrett's esophagus underwent monthly stepwise cryoablation with pressurized CO (2) gas, with follow-up esophageal biopsies until complete histological reversal was achieved. Responses were analyzed with an intention-to-treat analysis according to complete response for intestinal metaplasia (CR-IM), which was defined as the elimination of all intestinal metaplasia including specialized intestinal metaplasia (SIM), subsquamous SIM, and dysplasia with intestinal metaplasia in the biopsies under narrow-band imaging (NBI). RESULTS: In total, 22 patients were enrolled, 20 of whom completed the treatment. Two patients declined further ablation after the first cryotherapy session. A total of 44 sessions were performed; a median of 2 sessions per patient (range 1 - 3 sessions) were needed to complete the ablation of Barrett's esophagus. No severe complications occurred. Follow-up endoscopies were performed in 20 patients (90.9 %). Two patients (9.1 %) were lost to follow-up. Median follow-up was 10 months (range 6 - 18 months). After cryotherapy, 20 patients (90.9 %) reached CR-IM of Barrett's esophagus. Patients underwent a median number of 3 follow-up endoscopies (range 2 - 4) with biopsies. At 6 months, recurrence was evident in three patients (13.6 % of the overall population, 15.0 % of the CR-IM population). One of the three patients developed intestinal metaplasia but no dysplastic change and the other two developed subsquamous SIM. CONCLUSIONS: The pressurized CO (2) spray cryotherapy is a relatively effective and safe endoscopic treatment for Barrett's esophagus.

Matrix metalloprotease expressions in both reperfusion lung injury and oleic acid lung injury models and the protective effects of ilomastat.

OBJECTIVE: Our aim was to study the expressions of matrix metalloprotease 9 (MMP9) and the effects of the MMP inhibitor Ilomastat in both ischemia/reperfusion (I/R)- and oleic acid (OA)-induced lung injury models. MATERIALS AND METHODS: Real-time polymerase chain reactions and Western blots were used to assess mRNA and protein expressions of MMP9 in lung tissues after I/R or OA lung injury. Ischemia was induced by clamping one branch of the pulmonary artery for 60 minutes and then reperfusing for 120 minutes. In the OA model, lung injury was induced by intravenous infusion of OA (0.1 mL/kg) for 20 minutes and then observation for 6 hours. Lavage leukocyte concentration and wet/dry lung weight ratio were used to assess lung inflammation and injury. Blood samples were collected for assays of hydroxyl radicals and nitric oxide. The MMP inhibitor Ilomastat (100 microg/kg) was administered before I/R and OA infusion. RESULTS: mRNA and protein expressions of MMP9 were significantly increased in both lung injury models. Ilomastat decreased MMP9 mRNA and protein expressions but did not reach statistical significance. Blood concentrations of hydroxyl radicals and nitric oxide, wet/dry lung weight ratios, and lavage leukocyte concentrations were significantly higher in both experimental groups compared with the sham group (P < .001). Ilomastat significantly attenuated the extent of lung inflammation and injury induced by both I/R and OA. CONCLUSION: MMP may play a critical role in the lung injury induced by I/R and OA infusion.

The association between MIF-173 G>C polymorphism and prostate cancer in southern Chinese.

BACKGROUND AND OBJECTIVES: Accumulating epidemiological and molecular evidence suggests that inflammation is an important component in the etiology of PCa. Macrophage migration inhibitory factor (MIF) plays an important role in the pro- and anti-inflammatory response to infection. This study is aimed at investigating the potential association between MIF-173 G>C polymorphism, Gleason score, clinical stage, and prostate-specific antigen (PSA) value with respect to PCa incidence among the Han nationality in Southern China. METHODS: Genotyping was performed by using tetraprimer polymerase chain reaction (PCR) on 259 PCa patients and 301 cancer-free controls. RESULTS: We found that the MIF-173*C variant allele was significantly associated with an increased risk of PCa [adjusted odd ratio (OR) = 2.99, 95% confident interval (CI): 1.94-4.60] and higher Gleason scores from the PCa subjects (adjusted OR = 10.72, 95% CI: 5.35-21.49). In addition, we noted that the MIF -173*C variant allele was related to higher clinical stages and PSA values in PCa patients (adjusted OR = 15.68, 95% CI: 7.40-33.23; adjusted OR = 4.37, 95% CI: 2.41-7.92, respectively). CONCLUSION: Our data suggest that MIF-173 polymorphisms may be associated with a higher incidence of prostate cancer compared to controls, and appears to be associated with higher Gleason scores, higher clinical stages, and PSA values in those with prostate cancer.

Inducible nitric oxide synthase expressions in different lung injury models and the protective effect of aminoguanidine.

OBJECTIVE: Our aim was to study the expression of inducible nitric oxide synthase (iNOS) in 2 experimental models: (1) ischemia/reperfusion (I/R) of the lung tissues and (2) oleic acid infusion. The protective effect of an iNOS inhibitor, aminoguanidine, was evaluated in these 2 injury models. MATERIALS AND METHODS: Real-time polymerase chain reactions and Western blots were used to assess the mRNA and protein expressions of iNOS in lung tissues after applying 2 injury models. In the I/R model, ischemia was induced by clamping one branch of the pulmonary artery for 60 minutes and then reperfusing for 120 minutes. In the bone fracture model, lung injury was induced by intravenous (IV) infusion of oleic acid (0.1 mL/kg); analysis was performed 6 hours after injury. Blood samples were collected for the assay of 3 inflammatory parameters: tumor necrosis factor alpha, hydroxyl radicals, and nitric oxide (NO). The wet/dry lung weight ratio was used as a parameter reflecting the lung injury level. RESULTS: mRNA and protein expressions of iNOS were significantly increased in these 2 lung injury models compared with the controls. Blood concentrations of TNFalpha, hydroxyl radicals, NO, and wet/dry lung weight ratio were also significantly higher in the 2 experimental groups than in the sham-treated group. The iNOS inhibitor aminoguanidine (20 mg/kg) significantly attenuated the lung injury induced by these challenges. CONCLUSIONS: Reperfusion of the ischemic lung tissues or IV infusion of oleic acid can both induce lung injury by activating systemic inflammatory responses and inducing iNOS expression. Administration of aminoguanidine can significantly attenuate the injury, suggesting that iNOS expression may play a critical role in the lung injury induced in these 2 models.

Cholinergic neurons of mouse intrinsic cardiac ganglia contain noradrenergic enzymes, norepinephrine transporters, and the neurotrophin receptors tropomyosin-related kinase A and p75.

Half of the cholinergic neurons of human and primate intrinsic cardiac ganglia (ICG) have a dual cholinergic/noradrenergic phenotype. Likewise, a large subpopulation of cholinergic neurons of the mouse heart expresses enzymes needed for synthesis of norepinephrine (NE), but they lack the vesicular monoamine transporter type 2 (VMAT2) required for catecholamine storage. In the present study, we determined the full scope of noradrenergic properties (i.e. synthetic enzymes and transporters) expressed by cholinergic neurons of mouse ICG, estimated the relative abundance of neurons expressing different elements of the noradrenergic phenotype, and evaluated the colocalization of cholinergic and noradrenergic markers in atrial nerve fibers. Stellate ganglia were used as a positive control for noradrenergic markers. Using fluorescence immunohistochemistry and confocal microscopy, we found that about 30% of cholinergic cell bodies contained tyrosine hydroxylase (TH), including the activated form that is phosphorylated at Ser-40 (pSer40 TH). Dopamine beta-hydroxylase (DBH) and norepinephrine transporter (NET) were present in all cholinergic somata, indicating a wider capability for dopamine metabolism and catecholamine uptake. Yet, cholinergic somata lacked VMAT2, precluding the potential for NE storage and vesicular release. In contrast to cholinergic somata, cardiac nerve fibers rarely showed colocalization of cholinergic and noradrenergic markers. Instead, these labels were closely apposed but clearly distinct from each other. Since cholinergic somata expressed several noradrenergic proteins, we questioned whether these neurons might also contain trophic factor receptors typical of noradrenergic neurons. Indeed, we found that all cholinergic cell bodies of mouse ICG, like noradrenergic cell bodies of the stellate ganglia, contained both tropomyosin-related kinase A (TrkA) and p75 neurotrophin receptors. Collectively, these findings demonstrate that mouse intrinsic cardiac neurons (ICNs), like those of humans, have a complex neurochemical phenotype that goes beyond the classical view of cardiac parasympathetic neurons. They also suggest that neurotrophins and local NE synthesis might have important effects on neurons of the mouse ICG.

Reperfusion liver injury induces down-regulation of eNOS and up-regulation of iNOS in lung tissues.

OBJECTIVES: Acute lung injury and inflammation can occur after hepatic ischemia/reperfusion (I/R). Little is known regarding the possible role of nitric oxide synthase expression in this complex type of lung injury. METHODS: Real-time polymerase chain reactions and immunohistochemistry were used to assess the mRNA and protein expression of eNOS and iNOS in lung tissue after I/R challenge to the liver. Ischemia was induced by clamping the hepatic artery and portal vein for 40 minutes. After flow was restored, the liver was reperfused for 300 minutes. Blood samples were collected to assay three inflammatory parameters: tumor necrosis factor (TNF)-alpha, hydroxyl radicals, and NO. Lung lavage samples were assayed for protein and myeloperoxidase. The expression of eNOS and iNOS in lung tissues (n = 3) was also evaluated after I/R challenge to the liver. The iNOS inhibitor aminoguanidine was also tested in this I/R model. RESULTS: Reperfusion of the liver produced increased blood concentrations of TNF, hydroxyl radicals, and NO (P < .001; n = 8). Bronchial lavage fluids showed higher levels of protein and myeloperoxidase in the I/R than in the sham-treated group (P < .01). eNOS expression was down-regulated and iNOS expression up-regulated in I/R lung tissues (n = 3). The iNOS inhibitor aminoguanidine (10 mg/kg) significantly attenuated the lung injury. CONCLUSIONS: I/R injury to the liver induced lung injury involving systemic inflammatory responses and iNOS expression. Administration of aminoguanidine significantly attenuated the injury, suggesting that iNOS expression may play a critical role in lung injury induced by I/R of the liver.

Extraperitoneal laparoscopic radical prostatectomy.

A total of 28 patients with clinically localized prostate cancer (PCa) underwent extraperitoneal laparoscopic radical prostatectomy (EP-LRP). The mean operative duration was 309 (287-600) minutes. Estimated blood loss ranged from 380 to 1000 (mean 480) ml. At 3 to 5 days postoperatively, the catheter was removed. No open conversion was required and no patient presented postoperative complications. PSA level was less than 0.1 ng/ml at 3 months after surgery in all patients. At a mean follow-up of 10 (6-16) months, there were no biochemical failures. The extraperitoneal technique potentially decreased the risk of intra-abdominal complications and better approximated than open retropubic radical prostatectomy. In conclusion, EP-LRP is an effective, safe and precise technique.

Protective intestinal anti-rotavirus B cell immunity is dependent on alpha 4 beta 7 integrin expression but does not require IgA antibody production.

Rotavirus (RV) is the main cause of severe gastroenteritis in young children; protection has been correlated with intestinal Ab responses. Using a mouse model of RV infection and beta(7)-deficient (beta(7)(-/-)) mice, which do not express alpha(4)beta(7) integrin, we demonstrated the importance of alpha(4)beta(7) integrin in B cell-mediated anti-RV immunity. beta(7)(-/-) mice acutely infected with murine RV resolved infection and developed normal serum IgG Abs but had diminished intestinal IgA responses. alpha(4)beta(7)(-/-) immune B cells did not resolve RV infection when adoptively transferred into RV-infected Rag-2-deficient mice. Fewer RV-specific B cells were found in the intestine of Rag-2-deficient mice transferred with beta(7)(-/-) B cells compared with wild type. The absence of alpha(4)beta(7) expression and/or a lower frequency of IgA-producing cells among transferred beta(7)(-/-) B cells could have accounted for the inability of these cells to resolve RV infection following passive transfer. To distinguish between these possibilities, we studied the importance of IgA production in RV infection using IgA-deficient (IgA(-/-)) mice. IgA(-/-) mice depleted of CD8(+) T cells were able to clear primary RV infection. Similarly, adoptive transfer of immune IgA(-/-) B cells into chronically infected Rag-2-deficient mice resolved RV infection. We further demonstrated in both wild-type and IgA(-/-) mice that, following oral RV infection, protective B cells reside in the alpha(4)beta(7)(high) population. Our findings suggest that alpha(4)beta(7) integrin expression is necessary for B cell-mediated immunity to RV independent of the presence of IgA.

[Micro determination of acrylonitrile in ambient air by gas chromatography].

Acrylonitrile in ambient air was collected by Tenax GC and then thermo-desorbed. It was separated on GDX-502 chromatographic column from other pollutants and determined by flame-ion ization detector. The minimum detectable concentration was 0.01 mg/m3 when 2 L ambient air was collected. The recoveries were 85.6%-105.4% and RSDs were 4.5%-6.2%.

The interleukin-4/interleukin-13 receptor of human synovial fibroblasts: overexpression of the nonsignaling interleukin-13 receptor alpha2.

Interleukin (IL)-4 and IL-13 are known to bind to shared heteromultimeric receptor complexes of variable composition. Given the many regulatory effects of IL-4 and IL-13 on synovial cells, we aimed to characterize their IL-4/IL-13 receptor (R). Cultivated synovial fibroblasts expressed transcripts for IL-4Ralpha and IL-13Ralpha1, the human homolog of the recently cloned mouse IL-13R, but not the common gamma-chain of the IL-2R. In particular, IL-13Ralpha2 mRNA, encoding a different IL-13R recently cloned from human renal carcinoma cells, was expressed at a strikingly high level. Correspondingly, a predominant protein migrating at 65 to 75 kd was cross-linked by iodinated IL-13 and was not cross-competed by an excess of unlabeled IL-4. However, by flow cytofluorometry, IL-13Ralpha1 (detected by the anti-lL-13Ralpha1 mAb 65) and IL-4Ralpha (detected by the mAb S697) were expressed at similar low density. Radioligand binding studies revealed for both cytokines approximately 300 receptors/cell with similar high affinity. An additional class of IL-13Rs was identified after occupation of the shared high-affinity receptors by the nonsignaling, double-mutant IL-4121R-->D, 124Y-->D (RY-IL-4). In these experiments, 1251-IL-13 bound to a single receptor population with a Kd of approximately 300 pM and approximately 5000 sites/cell, matching the published affinity of monomeric IL-13Ralpha2 when expressed in COS7 cells. RY-IL-4 blocked the IL-4- and IL-13-mediated vascular cell adhesion molecule (VCAM)-1 expression and Stat6 activation, suggesting that the large number of high-affinity IL-13Ralpha2 monomers are silent receptors, likely representing a decoy target for IL-13.