The human GM CSF cDNAandEGFP cDNA were linked together with IRES and then inserted into the expression vector pCI Egr 1, which was constructed by substituting CMV promoter in pCIneo with the Egr 1 promoter(Egr EG).The vector was transfered into human bone marrow stromal cell line HFCL by Lipofectin TM . The results indicated that the activity of EGFP in transfected cells increased at 18h after exposure to 2.5 Gy. The amounts of secreted GM CSF and CFU GM in serum free supernatants of Egr EG was significantly higher than the control group. EGFP and GM CSF cDNA were successfully integrated and expressed in the cells, which were confirmed by FACS and RT PCR analysis respectively. These in vitro data provide an experimental basis for the in vivo use of gene therapy of GM CSF gene regulated by Egr 1 promoter to protect hematopoiesis from total body irradiation.

Objective To explore the chemotherapeutic agents-induced modulating effects of Egr-1 promoter sequence on GM-CSF expression and its protective effect against injury to haematopoiesis due to chemotherapy.Methods Human GM-CSF cDNA and enhanced green fluorescent protein(EGFP) cDNA were linked to internal ribosome entry site(IRES) respectively,and then recombined to pCIneo vector containing Egr-1 promoter(Egr-EG).The vector was transferred into human bone marrow stromal cell line HFCL by lipofection,and the HFCL/EG cells were then finally constructed.MTT assay was performed to determine the effects of cisplatin(DDP),5-fluorouracil(5-FU),gemcitabine(GEM) and paclitaxel(PTX) on the survival rates of HFCL/EG and HFCL cells,and the IC50 of such agents against HFCL/EG and HFCL cells were calculated.The percentage of HFCL/EG cells which positively expressed EGFP was assessed by both flow cytometry and inverted fluorescence microscope.The effects of the active oxygen inhibitor N-acetylcysteine(NAC) on the expression of GM-CSF,which was modulated by promotor Egr-1,were detected by ELISA.The effects of HFCL/EG supernatant on CFU-GM after being exposed to chemotherapeutic agents were examined.Results With different sensitivity to DDP,5-FU,GEM and PTX,HFCL/EG cells were successfully constructed.The drugs showed higher IC50 value against HFCL/EG cells than HFCL cells(P

Objective To investigate the effects of sevoflurane anesthesia in diabetic pregnant rats on the cognitive function of the offspring rats. Methods Forty female Sprague?Dawley rats and 5 male rats, weighing 200-250 g, were used in the study. Twenty pregnant rats at 7 weeks of gestation were randomly selected, and diabetes mellitus was induced by intraperitoneal streptozotocin 45 mg∕kg and confirmed by blood glucose level>10.4 mmol∕L. Twenty pregnant rats at 20 days of gestation, in which diabetes mellitus was not induced, were selected and divided into 2 groups ( n=10 each) using a random number table:sevoflurane group (group S) and control group (group C). Twenty pregnant rats at 20 days of gestation with diabetes mellitus were selected and divided into 2 groups ( n=10 each) using a random number table:sevoflurane group (group DS) and control group ( group DC). In DS and S groups, the pregnant rats were placed in a self?made anesthetic box and inhaled 2% sevoflurane for 2 h. At 6 weeks after birth, the offspring rats were selected, and Morris water maze test was performed. The rats were sacrificed, brains were removed, and the hippocampi and cortex were removed for determination of phosphorylated cyclic a?denosine monophosphate response element?binding protein ( p?CREB) expression using immuno?histochem?istry. Results Compared with group C, the escape latency was significantly prolonged, and the frequency of crossing the original platform was significantly decreased in S and DC groups ( P<0.05) . Compared with group DC, the escape latency was significantly prolonged, and the frequency of crossing the original plat?form was significantly decreased in group DS (P<0.01). Compared with group S, the escape latency was significantly prolonged, the frequency of crossing the original platform was significantly decreased ( P<0.05) , and lighter staining for p?CREB was found, and the number of p?CREB positive cells was decreased in the hippocampus and cortex in group DS. Conclusion Sevoflurane anesthesia?induced cognitive dys?function is aggravated in the offspring rats of diabetic pregnant rats, and the mechanism is related to inhibi?tion of CREB phosphorylation.

Objective To explore the diagnostic value of subacute thyroiditis by fine needle aspiration cytology(FNAC).Method 3665 patients with thyroid disease were detected by FNAC method in 8 years(from Feb 1990 to Dec 1998).Results 353 cases(9 63%) of subacute thyroidits were definitely diagnosed by FNAC with vital staining.Conclusions As compared with other clinical examination including CT,ultrasonic B and radioimmunological assay.FNAC test is all the more simple,safe and valuable diagnostic method for subacute thyroiditis.

Objective To summarize the clinical features of uncinate process carcinoma of the pancreas and improve the resection rate. Methods From Jan. 1990 to Jun.1999, Ten cases of pancreas uncinate process carcinoma received Whipple′s procedure. Portal vein resection and reanastomosis was performed in 5 cases, among them the resected length varied between 20～42 *!cm.Two cases underwent PV lateral wall partial resection. The longest time of PV blood occlusion was 40 *!minutes.Results Among 7 cases undergoing PV resection, 1 died with hepatic failure 3 days post-op. 1 case suffered postoperative chylous ascites.These 6 cases survived 13 to 29 months post-op. Among 3 cases not undergoing PV resection, 2 cases survived 13 months and 14 months, respectively. 1 case is still alive at the follow-up of 115 months post-op. Conclusion Pancreas uncinate process carcinoma has the tendency of invading adjacent PV and SMV, which should not be simply regarded as a contraindication of the radical resection.

0.05).Conclusions Immunologic function alteration of MPP of infants is showed mainly that Th1 immunologic response is raised,Th2 immunologic response is abated,the cellular factor are changed horizontally,but the immunology index and cellular factor are recovered quickly.There are no relations between MPP of infant and PHF-AH genotype,but there is a downward trend in activity of PHF-AH in acute stage.

Objective To evaluate the effect of sleep deprivation on cognitive function in the rats undergoing propofol anesthesia.Methods Sixty healthy male Sprague Dawley rats,aged 14-18 weeks,weighing 200-250 g,were randomly assigned into 3 groups (n=20 each) using a random number table:control group (group C),propofol anesthesia group (group P),and sleep deprivation + propofol anesthesia group (group SDP).Propofol was given as a bolus of 15 mg/kg followed by an infusion of 40 mg · kg-1 · h-1 for 2 h in group P.After the rats were subjected to rapid eye movement sleep deprivation for 24 h,the rats received propofol anesthesia in group SDP.Before sleep deprivation,after sleep deprivation,and at 1,3 and 7 days after anesthesia,Morris water maze test was used to assess the learning and memory function,and the escape latency and frequency of crossing the original platform were recorded.Ten rats randomly selected from each group at 1 and 7 days after anesthesia were sacrificed,and brains were removed to observe the morphology of nerve cells in the hippocampal CA1 region (by Nissl's staining) and to detect the expression of phosphorylated Tau at Thr231 (Tau-pThr231) in the hippocampal CA1 region (by immunohistochemisty).Results Compared with group C,the escape latency was significantly prolonged,the frequency of crossing the original platform was decreased,the expression of Tau-pThr231 in the hippocampal CA1 region was up-regulated at 1 day after anesthesia in P and SDP groups (P＜0.05),especially in group SDP (P＜0.05),and there was no significant difference between the groups at the other time points (P＞0.05).The pathological changes were aggravated at 1 day after anesthesia in group SDP compared with group P,and there was no significant difference at 3 and 7 days after anesthesia between group SDP and group P.Conclusion Sleep deprivation can aggravate the transient cognitive dysfunction after propofol anesthesia,and the mechanism is related to promotion of Tau phosphorylation in the rats.

Diagnosis, Differential ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Male ; Prostatic Hyperplasia ; diagnosis ; Prostatic Intraepithelial Neoplasia ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Racemases and Epimerases ; analysis ; Staining and Labeling ; methods

Objective To explore the effects of penehyclidine hydrochloride (PHC) on lung epithelial type Ⅱcells against oxidative stress damage. Methods A549 cells treated with H2O2 were used as oxidative stress damage cell model. A549 cells were divided into 3 groups: control group (C group), H2O2 treated group (H group) and PHC treated group (P group). The viability of A549 cells was measured by MTT assay. The apoptotic rate was measured by TUNEL assay. The lev?els of malonicdialed (MDA), reactive oxygen species (SOD), reduced glutathione hormone (GSH) and nicotinamide adenine dinucleotide phosphate (NADPH) of cells were detected by biochemistry colorimetry. Results Compared with group C, the viability of A549 and the contents of SOD, GSH and NADPH were significantly decreased in group H, while MDA content and apoptotic rate were increased (P<0.05). Compared with group H, the viability of A549, the contents of SOD, GSH and NADPH were significantly increased in group P, while MDA content and apoptotic rate were reduced (P<0.05). Conclu?sion Penehyclidine hydrochloride shows protective effects on A549 cells through reducing the oxidative damage induced by H2O2.

Objective To investigate the association of ERCC1 and BRCA1 gene expression with clinical features and sensitivity to platinum-containing chemotherapy in patients with ovarian epithelial carcinoma. Methods Primary ovarian epithelial carcinoma tissues were harvested from 48 patients receiving staging surgery or cytoreductive surgery. Expression of ERCC1 and BRCA1 in the tumor samples was detected by immunohistochemistry. Results ERCC1 expression was correlated with clinical stage(P﹤0. 05)but not with age,pathological type or degree of differentiation(P﹥0. 05). BRCA1 expression was not correlated with any of the clinicopathological features(P﹥0. 05);ERCC1 expression was significantly higher in drug-resistant tissues than in drug-sensitive samples(P﹤0. 05);The expression of ERCC1 and BRCA1 was positive in 89. 58%and 25. 00%of the samples,respectively. Conclusion ERCC1 gene expression is correlated with clinical stage but not with age,pathological type or degree of differentiation. BRCA1 gene expression is not correlated with clinicopathological features. ERCC1 has high positive expression in epithelial ovarian cancer and is correlated with sensitivity of platinum-containing adjuvant chemotherapy.