Objective To investigate the clinical significance of WT1 gene expression in the urine. Methods The expression of WT1 gene in the urine of patients with chronic glomerular nephritis(CGN), rheumatic diseases,diabetes mellitus(DM) and healthy subjects was detected by semi-nested PCR. Results The positive rates of WT1 gene expression in CGN with proteinuria,DM with proteinuria, DM without proteinuria and rheumatic diseases were 46.7% (14/30),44.4% (16/36),5.95% (5/84) and 16%(4/25), respectively. WT1 expression in the urine was not related to the degree of proteinuria, hematuria, renal function or clinical course of diseases. WT1 expression in the urine of the healthy subjects or cystitis patients was negative. Conclusion The measurement of WT1 mRNA in the urine could play an important role in early diagnosis of progressive renal damage.

Objective To investigate the association of anti-C1q antibody with disease activity and lupus nephritis in patients with systemic lupus erythematosus(SLE). Methods The serum Level of anti-C1q antibody was determined in 35 patients with SLE by ELISA. The correlation between anti-C1q level and other disease activity parameters, such as SLEDAI score, anti-dsDNA antibody, C3, C4 and CH50 levels, was analyzed. Results Anti-C1q antibody was positive in 51.4% of patients with SLE, the positive rates in patients with active nephritis and without one was 77.8%(14/18) and 42.3%(22/52), respectively, and that was significantly higher in the former than in the later(P

Objective To study the clinical significance of neuron specific enolase (NSE),carcinoembryonic antigen(CEA) and carcinoma associated antigen(CA125) in diagnosis,evaluation of therapy and monitoring metastasis and recurrence of lung cancer.Methods The levels of NSE,CEA and CA125 were detected in the sera from 30 healthy controls,48 patients with benign diseases,50 patients with non-small cell lung cancer(NSCLC) and 14 patients with small cell lung cancer(SCLC) by using microparticle enzyme linked immunoassay and bioantibodies sandwich one step assay by using streptavidin technique,respectively.Results The levels of NSE and CEA in the sera of patients with NSCLC and SCLC were significantly higher than those of healthy controls and patients with benign diseases (both P0 05).The levels of NSE CEA and CA125 decreased after the treatment and increased greatly with the metastasis and recurrence of the cancer.After the combination of NSE and CEA and CA125,the sensitivity significantly increased,but no difference was found in the specifity.Conclusion Measuring serum levels of NSE,CEA and CA125 are very useful in diagnosis,evaluation of therapy and monitoring of metastasis and recurrence of lung cancer.

Objective To investigate the association of the expression level of anti-apoptosis protein p-AKT and gastric mucasal injury induced by indomethacin in mice.Methods The cytotoxicity induced by indomethacin was measured by LDH assay.The p-AKT expression levels were measured in the gastric mucosal tissues from C57BL/6 mice and rat gastric mucosal (RGM-1) cell lines treated with indomethacin lay western blotting.Results The cytotoxicity induced by indomethacin was in a dose dependent manner.Compared with the control,a typical histological appearance of gastric ulcer was observed in the gastric mucosa of in domethacin-administered mice;p-AKT protein expression in the gastric mucosa of mice and RGM-1 cell lines was decreased after treated with indomethacin.Conclusion The reduction of Anfi-apoptesis protein p-AKT expression may be a new mechanism for the gastric mucosal injury induced by indomethacin.

Objective To investigate the relationship among the concentration of plasma interleukin (IL)-10 and plasma IL-13 and the clinical therapeutic effect in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods Thirty-six AECOPD inpatients were enrolled in this study.Blood samples of the subjects were collected as soon as hospitalization, and plasma IL-10 and IL-13concentrations were analyzed by enzyme-linked immunosorbent assay.The clinical manifestations of subjects were quantified by a special designed score standard, and were evaluated at the time points of hospitalization, 48 hours treatment and 96 hours treatment.The therapeutic effect was evaluated by clinical manifestations score combined with pulmonary ventilation functional parameter.Eventually, the correlation among the concentration of IL-10 concentration and IL-13 and the clinical therapeutic effect were analyzed.Results The correlation coefficients between clinical manifestations score decrease and pulmonary ventilation function improvement with plasma IL-10 concentration after 48 hours treatment were 0.85 and 0.48 respectively,then, 0.64 and 0.52 after 96 hours treatment.The correlation coefficients between clinical manifestations score decrease and pulmonary ventilation function improvement with plasma IL-13 concentration after 48hours treatment were -0.41 and -0.34, after 96 hours treatment , correlation coefficients between clinical manifestations score decrease with plasma IL-13 concentration was -0.36.All of the above correlation coefficients were statistically significant.Conclusion Plasma IL-10 concentration was positively, whereas, IL-13 concentration was negatively correlated with the therapeutic effect in AECOPD patients.

BACKGROUND:The dynamic changes of hypoxia inducible factor-1 alpha (HIF-α) and inducible nitric oxide synthase (iNOS) genes in the pulmonary artery wall during the process of hypoxic pulmonary hypertension (HPH) development need to be investigated.OBJECTIVE: This study was to observe the gene expressions of HIF-α and iNOS in the pulmonary artery wall at the different hypoxic time points, and to investigate their effects in HPH development.DESIGN: Controlled observation animal experiment.SETTING: Department of Respiration, Second Hospital Affiliated to Nanhua University.MATERIALS: Forty healthy male Wistar rats of clean grade, aged 6-8 weeks, with body mass of (220±10) g were involved in this study. They were randomized into control group (n =8) and hypoxia group (n =32). Four time points, i.e.3, 7, 14, and 21 days after hypoxia were set for the animals in the hypoxia group, 8 rats at each time point.METHODS: This study was carried out in the Institute of Oncology, Nanhua University between August 2004 and December 2005. Rats in the hypoixa group were treated according to the method reported by Li et al. Hypoxia treatment was omitted for the rats in the control group. At each time point, the rats were anesthetized, and then a micro-catheter was inserted into the right jugular vein and connected to a multichannel physiologic recorder, which was used for detecting the mean pulmonary arterial pressure (mPAP). The heart of each euthanized rat was taken out, and its right ventricle (RV), and left ventricle and septum (LV+S) were weighted. Right ventrical hypertrophy index (RVHI) reflected right ventricle hypertrophy degree. Right upper lung tissue of rat was harvested for haematoxylin & eosin (HE) staining and elastic fiber staining. Pathological image analysis software was used to determine pulmonary arterial wall area/total vascular area, lumina area/total vascular area, smooth muscle cell density in the media of pulmonary arteriole, and media thickness of pulmonary arteriole, which were used as remodeling indexes of pulmonary arteriole. HIF-1α and iNOS in the pulmonary arteriole were performed in situ hybridization and immunohistochemical detection. The mean absorbance of pulmonary arteriole wall was used as the relative content of mRNA expression and protein level.MAIN OUTCOME MEASURES: ①Changes in mPAP, RV hypertrophy degree and remodeling indexes of pulmonary arteriole of rats. ② Expressions of HIF-α and iNOS as well as their correlations with mPAP and remodeling indexes of pulmonary arteriole.RESULTS: All the 40 Wistar rats were involved in the final analysis. ①At hypoxia 7 days, mPAP was significantly higher than that of control group (P ＜ 0.05). mPAP reached to the high level at hypoxia 14 days, and then maintained at this level. ② At hypoxia 14 days, RVHI was higher than that of control group (P ＜ 0.05). ③ At hypoxia 7 days, pulmonary arteriole wall was thickened, and lumina became narrowed. There were significant differences in pulmonary arterial wall area/total vascular area, lumina area/total vascular area between hypoxia group and control group (P ＜ 0.05). At hypoxia 14 days, smooth muscle cell density in the intima-media of pulmonary arteriole, and intima-media thickness of pulmonary arteriole were significantly increased (P ＜ 0.05). At hypoxia 21 days, lumina was further narrowed, and obvious smooth muscle hyperplasy was found. HIF-1α and iNOS mRNA expression presented weak-positive in the control group; The relative content of HIF-1α mRNA did not significantly alter at hypoxia 3 and 7 days, but was obviously increased at hypoxia 14 days ,and after this, it maintained at high level. iNOS mRNA was markedly higher than that in the control group at hypoxia 3 days, reached its peak at hypoxia 7 days, was close to the level in the control group at hypoxia 14 days, and enhanced again at hypoxia 21 days, but it was still lower than that at hypoxia 3 days. ⑤HIF-α was mainly found in the intima and media, while iNOS was found in the whole layer of vessels. iNOS was weakly expressed in the intima and media of pulmonary vessels in the control group. No obvious difference in iNOS existed at hypoxia 3 days as compared with control group. iNOS was obviously expressed in media and intima at hypoxia 7 days. Vascular media was thickened at hypoxia 14 days, and the expression of iNOS was enhanced. iNOS was not found in the vascular adventitia in the control group, but was found in the vascular adventitia in the hypoxia group. ⑥mPAP was positively correlated with pulmonary vascular remodeling (r =0.976, P ＜ 0.01 ), and HIF-1α mRNA was positively correlated with iNOS protein(r =0.927, P ＜ 0.05).CONCLUSION: Both HIF-1α and iNOS exert effects in the process of HPH development of rats, and HIF-1α and iNOS gene expression may be mutually regulated.

Objective To compare three rat models of acute stress response and to explore an ideal experimental rat model for research of acute stress response .Methods A total of 40 clean grade male SD rats were randomly ( by random number ) divided into 4 groups ( n=10 each ):Normal group ( group I ) , caudal trunk transection group ( group II), burn group (group III), and amputation group (group IV).The group I received no special treatment , the group II received inferior caudal trunk transection at the level between sacral spinal nerve 3 and 4 (S3, S4), the group III was inflicted with 30%total body surface area ( TBSA) grade 3 burn on the back , and the group IV had an amputation of the left lower limb as severe traumatic stress .Rats in each group were killed at 30 minutes after treatment and blood samples were collected for measuring serum adrenocorticotropic hormone (ACTH) and corticosterone (CORT) levels by ELISA. The c-fos protein expression in hypothalamic tissue was examined by immunohistochemistry .Results Compared with the group I, serum ACTH and CORT levels , and hypothalamic expression of c-fos protein in the group II , III, IV were significantly increased ( P <0.05 ) .Compared with the group III , serum ACTH and CORT levels , and hypothalamic expression of c-fos protein were significantly higher in the group II and IV ( P <0.05 ) , while there was no significant difference between the group II and IV (P>0.05).Conclusions The preparation of acute stress response model induced by inferior caudal trunk transection has simple operation steps and produces a traumatic injury to a similar degree , and quite well reflects the acute stress response in humans caused by sudden accident .Therefore it is a quite good method to establish acute stress response and deserves further investigation .

Objective To investigate the influence of electroacupuncture(EA) in different points combined with spinal anesthesia on acute stress response and mechanisms related with expression of spinal c-fos protein. Methods Fifty adult male Sprague-Dawley rats were randomly divided into 5 groups:groupⅠwere normal rats receiving only 50μL saline intrathecal IT injection;groupⅡwere docked rats receiving IT with 50 μL saline;groupⅢ were docked rats receiving IT injection of 50 μL, 2.5 μg fentanyl; group Ⅳwere docked rats receiving IT injection of fentanyl 50μL and EA at′zu-san-li′;groupⅤwere docked rats receiving EA at′er xue′and IT fentanyl. Serum corticosterone (CORT) and adrenocorticotropic hormone (ACTH) levels were measured by ELISA. Determined the c-fos protein expression levels of L4-6 spinal by western blotting. Result GroupⅡACTH, CORT levels and c-fos expression was significantly higher than those in groupⅠ(P<0.01). Compared with groupⅡ, groupⅢ,ⅣandⅤhad higher ACTH and CORT levels (P<0.05). However, c-fos expression in the spinal cord between groupⅡandⅢshowed no difference (P>0.05). Between groupⅢandⅣ, no differences in serum hormone levels (P > 0.05) were detected. In group Ⅴ the c-fos expression was lower than that of the Ⅳ group (P < 0.01). Conclusion Body acupuncture and auricular electrical stimulation combined with intrathecal anesthesia were effective to reduce pain in rats with acute stress reactions. Under intrathecal anesthesia, auricular stimulation played a better role of non-nerve block region, and the mechanism may be due to the release of neurotransmitters induced c-fos protein in spinal cord.

Objective To explore the effect of electroacupuncture (EA) at tusanli (ST36) on regulation of stress response under different doses of etomidate anesthesia in rats.Methods Sixty male Sprague-Dawley rats, weighing 280-310 g, were randomly divided into control group (group C), model group (group M), etomidate 60 mg/kg group (group E1), etomidate 30 mg/kg group (group E2), etomidate 60 mg/kg combined with EA group (group EA1) and etomidate 30 mg/kg combined with EA group (group EA2), n=10 in each group.All groups received inferior caudal trunk transection at the level between sacral spinal nerve 3 and 4 (S3, S4) to prepare acute stress response model except group C.Group M received no others treatment.The rats in group E1, group EA1, group E2 and group EA2 were intraperitoneally injected with 60, 60, 30 and 30 mg/kg etomidate, respectively.Group EA1 and group EA2 received EA ST36.The points were stimulated at a frequency of 2/100 Hz with 1 mA output and a dilatational wave, which lasted for 30 min.ACTH and Cor levels were measured by ELISA.The c-fos protein expression in hypothalamic tissue was examined by Western blot.Results Compared with group C, ACTH and Cor levels, and hypothalamic expression of c-fos protein in group M were significantly increased (P<0.05).Compared with group M, serum ACTH and Cor levels, and hypothalamic expression of c-fos protein in groups E1, E2, EA1 and EA2 were significantly decreased (P<0.05).Compared with group E1, serum ACTH and Cor levels, and hypothalamic expression of c-fos protein were significantly higher in groups E2 and EA1 (P<0.05).Compared with group E2, serum ACTH level and hypothalamic expression of c-fos protein were significantly lower in group EA2 (P<0.05).Conclusion EA at ST36 regulating stress response under etomidate anesthesia in rats is effective and two-way, and the mechanism may be due to the release of neurotransmitters induced c-fos protein in hypothalamus.

Objective There is a lack objective methods for the diagnosis of postoperative cognitive dysfunction (POCD).This study aimed to investigate the influence of propofol or etomidate anes-thesia on the postoperative cognitive function and serum S 100βprotein level in infants . Methods This study included 100 hernia infants aged 1-3 years treated by laparoscopic herniorrhaphy under propofol (n=50) or etomidate anesthesia (n=50).At 1 day before and 3 days after surgery, we assessed the cognitive function of the patients using Bayley Scales of Infant and Toddler Development ( BSID-Ⅲ) and further divided each group into a POCD and a non-POCD sub-group based on the results of diagnosis made according to the Z-scores.Using ELISA, we measured the levels of the serum S100βpro-tein in the iliac venous blood drawn preoperatively ( T0 ) and before PACU ( T1 ) and compared them between the POCD and non-POCD groups. Results At 3 days after operation, POCD was observed in 10 cases (20.0%) in the propofol group and 9 cases (18.0%) in the etomidate group, with no statistically significant differences between the two (P>0.05).The level of the serum S100βprotein was markedly elevated in both the propofol and etomidate groups at T 1 as compared with that at T0(P<0.05), and so was it in the POCD in comparison with that in the non-POCD group (P<0.05), with no statistically significant differences between the two groups at T1(P>0.05).A significant correlation was found between the postoperative S 100βlevel and POCD at 3 days after surgery in both the propofol (r=0.842, P=0.001) and the etomidate group (r=0.821, P=0.001). Conclusion Propofol and etomidate anes-thesia can induce different degrees of postoperative decline of cognitive function in 1-3 years old infants .The post-anesthesia elevation of the serum S100βprotein level is positively correlated with early postoperative POCD and indicates various degrees of brain damage .