Aged ; Female ; Humans ; Male ; Middle Aged ; Silicosis

"CapFinder" technology,which can be used to clone the full length of 5′ UTR sequence of mRNA,was described.This technology used the terminal transferase activity of certain MMLV RT variants that added 3-5 residues(predominantly dC) to the 3′end of the first-strand cDNA exhibited when MMLV RT reached the 5′cap structure of mRNA.In the reverse reaction system containing GGG oligo,the terminal transferase activity was harnessed by the GGG oligo whose terminal stretch of dG residues can anneal to the dC-rich cDNA tail and serve as an extended template for RT.After RT switch templates from the mRNA template to the GGG oligo,a complete cDNA copy of the original RNA was synthesized with the additional GGG oligo sequences at the end.5′UTR of mRNA can be amplified with GGG oligo as forward primer and a gene-specific reverse primer.5′UTR of Bt toxin receptor E-Cadherin gene in midgut of cotton bollworm was cloned.

Objective To study the effects of Lipoxins A4(LXA4)on the expressions of aquaporin(AQP)1,3,5 in type Ⅱ pneumonocytes(ATⅡ)of rat treated with lipopolysaccharide(LPS).Method One pathogenfree male Spree Dawley(SD)rat every time.weighing 200～250 g,were used for the study.The typeⅡpenumonocytes of rats were isolated and purified,and the changes of cellular ultrastructure were observed by electron microscope in order to get the purity quotien＞90%.The type Ⅱ pneumonocytes were divided randomly into five groups,namely,vebicukun group(alcohol 0.7μL/mL),control group,LXA4 group(1×10-7mol/mL),endotoxin group(LPS 1μg/mL)and LXA4+LPS group(LXA4 1×10-7mol/mL,LPS 1μg/mL).AQP-1,3,5 mRNA of in the typeⅡpenumonocytes were assayed by using reversal transcription poly chain reaction(RT-PCR),and the expressions of AQP-1,3,5 protein were detected by using.immunohistochemistry(IHC).One each specimen,these tests were repeated for six times.ANOVA was used for statistical analysis.Results RT-PCR and IHC showed that when AT Ⅱ treated with 1 μg/mL LPS for 4 hours,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were significantly decreased in LPS group compared with control group(P＜0.01).However,the AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein after application of LXA4 significandy increased in LPS+LXA4 group in comparison with LPS group(LPS+LXA4,AQP1:0.647±0.132,AQP3:0.900±0.856,AQP5:0.879±0.058;LPS,AQP1:0.297±0.133,AQP3:0.512±0.113,AQP5:0.647±0.110;P＜0.01).The AQP-1,3,5 mRNA and the expressions of AQP-1,3,5 protein were aignificandy increased in LXA4 group in comparison with control group(LXA4,AQP1:0.539±0.142,AQP3:0.818 4-0.176,AQP5:0.841±0.066;Blank Control,AQP1:0.518±0.139;AQP3:0.138±0.136,AQP5:0.766±0.066;P＜0.01).Conclusions AQP-1,3,5 exist in typeⅡpenumonoeyte of rata,and the LXA4 can up-regulate the mRNA and protein expressions of AQP-1,3,5 in Type Ⅱ penumonocytes of rats treated with LPS.

AIM:To explore the effects of small interfering RNA(siRNA) targeting PCNA gene on nasopharyngeal carcinoma CNE2 cells growth and cycle.METHODS:Three synthesized siRNA targeting PCNA gene was transfected into CNE2 cells by using LipofectamineTM reagent.The PCNA mRNA and PCNA protein were detected by real-time polymerase chain reaction(RT-PCR) and immunohistochemical method.Inverted phase contrast microscope was used to determine the CEN2 cells growth before and after PCNA-siRNA transfected.Flow cytometry was used to observe the cell cycle.RESULTS:In CNE2 cells after PCNA-siRNA transfection,the expressions of PCNA mRNA and protein were down-regulated at different degree.Inhibition ratio of PCNA mRNA was 98.5%.Meanwhile,the cell cycle was suffocated at G0/G1 stage.CONCLUSION:The synthesized PCNA-siRNA effectively interferes nasopharyngeal carcinoma cells by down-regulating the expressions of the PCNA mRNA and its protein,therefore inhibits the growth of CNE2 cells.Future application of PCNA-siRNA in the gene therapy of nasopharyngeal carcinoma might be expected.

Objective To explore the value of laparoscopic diagnosis and treatment for Meckel’s diverticulum in children. Methods Laparoscopic exploration was performed in 11 children with suspected intestinal Meckel’s diverticulum. In other 2 children, the diverticulum was encountered during laparoscopic appendectomy, and then the umbilical incision was prolonged to take out the corresponding intestine for resection. Results No conversions to laparotomy were required in all the 13 children. The operation time lasted 30~90 min (mean, 75 min). The children were discharged at 5~6 postoperative days without complications. Conclusions Laparoscopic diagnosis and treatment for Meckel’s diverticulum in children is effective.

Objective To study the relation between copper,copper-zinc ratio in blood and blood-fat only in the copper intake lower than the normal acceptable daily intake,and to provide a unified dosage evidence for copper multi-ways exposure.Methods Forty-two SD rats were divided into 7 groups randomly.The rats in one group among them as the normal control group were fed on the normal fodder and water,the other 6 groups were fed on special fodder without copper and deionized water and i.g.copper gluconate at doses of 0ADI(the acceptable daily intake of rat),1/625ADI(0.000 128 mg/ml),1/125ADI(0.000 64 mg/ml),1/25ADI(0.003 2 mg/ml),1/5ADI(0.016 mg/ml) and ADI(0.8 mg/ml),for 30 days.The activity of ceruloplasmin(CP),the content of copper,zinc in the blood and the level of total cholesterol(TC),triglyceride(TG),high-density lipoprotein(HDL) and low density-lipoprotein(LDL) were determined.Results In the case of copper intake lower than the normal acceptable daily intake,the activity of CP in the blood of all treated rats were lower than the normal level(P

Hand Disinfection ; Health Personnel ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal

Objective To investigate the effectivity of bispectral index(BIS)and auditory -evoked potentials index(AEP index)in monitoring the depth of desflurane anesthesia in the elderly. Methods Sixty patients(ASA gradeⅠ～Ⅱ)without obvious auditory and psychiatric dysfunction undergoing general anesthesia were enrolled in this study,and they were devided into the elderly group (n=30)and the non-elderly group(n=30)according to the age.After the endotracheal intubation,the lungs were ventilated with desflurane in oxygen.End-tidal concentrations of desflurane were maintained at 0.6,1.0 minimum alveolar concentrations(MAC)and 1.3 MAC for 20 minutes, respectively.The changes of BIS,AEP index,mean arterial blood pressure(MAP)and heart rate (HR)were recorded simultaneously.Results In the elderly and non-elderly groups,BIS were 94.14?6.5 and 94.5?4.6(P＞0.05),and AEP index at pre-anesthesia were 87.2?11.7 and 89.2?6.9(P＞0.05)respectively,which showed no statistic difference between two groups.BIS and AEP index decreased significantly in each time point at pre-anesthesia compared with those during anesthesia in two groups (P＜0.05).In the periods of increasing concentration of desflurane,BIS decreased gradually in two groups(P＞0.05),but BIS in the elderly group were higher than in the non-elderly group at the same end-tidal desflurane concentration(P＞0.05),and BIS negatively correlated with the end-tidal desflurane concentration in two groups.In the elderly group,AEP index decreased significantly at the concentration of 1.0 MAC and 1.3 MAC of desflurane compared with that of 0.6 MAC(P＜0.05),but there was no difference in AEP index with increasing end-tidal desflurane concentration from 1.0 MAC to 1.3 MAC(P＞0.05).At every concentration of desflurane,AEPindexintheelderlygroupwaslessthaninthenon-elderlygroup(P＜0.05).Conclusions BIS and AEP indexes correlate well with the end-tidal desflurane concentration,and are valuable for monitoring the depth of desflurane anesthesia in the elderly.AEP index can predict the difference in depth of anesthesia at the same end-tidal desflurane concentration between the elderly and the non-elderly.

Objective To explore the therapeutic mechanisms of nitric oxide donor (SIN 1) in ischemic acute renal failure (IARF). Methods The effects of SIN 1 on intercellular adhesion molecule 1 (ICAM 1) expression and inflammatory cell infiltration in rat kidney after ischemia/reperfusion (I/R) injury were examined using ICAM 1, ?2 integrin polyclonal antibody, and immunohistochemistry. The renal functions were measured simultaneously. Results Progressive increased expression of ICAM 1 was found in kidneys, especially in vasa recta, after IR injury. ?2 integrin positive cells increased simultaneously in the outer medulla. The two indexes reached the peak values at 24 h after I/R injury. SIN 1 infusion at the beginning of reperfusion could remarkably inhibit the enhanced ICAM 1 expression, reduce the local infiltration of inflammatory cells, and ameliorate the renal functions. Conclusion NO donor can inhibit the renal expression of ICAM 1 and the infiltration of leukocytes after ischemia reperfusion injury, which may play an important role in the treatment of ischemic acute renal failure.

Objective To investigate the protective effect of genistein on endotoxin-induced acute lung injury in rats,and explore the underlying mechanisms.Methods Thirty-two male Sprague-Dawley rats were randomly divided into four experimental groups: saline control(S), genistein alone(G),lipopolysaccaride(LPS) alone(L),and genistein pretreatment(G+L).Each treatment group consisted of eight animals.Animals were observed for 6h after LPS challenge,and the wet/dry(W/D) weight ratio of the lung and the protein contents of the bronchoalveolar lavage fluid(BALF) were used as indexes of lung injury.Neutrophil recruitment and activation were evaluated by BALF cellularity and myeloperoxidase(MPO) activity.RT-PCR analysis was performed with lung tissue to assess the gene expression of ICAM-1.The histopathologic changes were also observed using the H&E stains of lung tissue.Results Lung injury parameters,including the wet/dry weight ratio and protein content in BALF,were significantly higher in the L group than in the S group(P