Histocytochemistry ; methods ; Humans ; Pathology, Clinical ; methods ; Specimen Handling ; methods ; Staining and Labeling ; methods

Immunohistochemical technique was an essential tool of conventional diagnosis,therefore,the application of immunohisto- chemistry in reformation of pathology laboratory teaching would boost pathological experimental teaching standards to a higher level.

Objective To investigate the distribution of water fluoride and the status of water-improving defluoridation projects,thus to explore the condition of endemic fluorosis in Guangxi Province.Methods According to"The National Technical Scheme for Endemic Disease Contml in 2005",the fuorine content in water Was determined by F-ion selective electrode,children's dental fluorosis was checked by Dean method.and the skeletal fluorosis was checked by the standard of clinical scale of skeletal fluorosis.Results 305 water samples in 61 villages were examined,among which 71 waters were exceeded the standard,accounting for 23.28%(71/305).The projects of defluoriding drinking water were running well except one was discarded.The prevalence of dental fluorosis was 13.55%(356/2627),the prevalence of skeletal fluorosis was 4.02%(65/1615).Conclusions The situation of endemic fluorosis control is not optimistic in Guangxi,which needs fuaher prevention and controls.

Objective To investigate the effect of hydrogen sulfide(H2S)on gene expression of adenosine triphosphate(ATP)-sensitive K+ channel(KATP)of aortic smooth muscle cells(ASMC)in rat.Methods Male Sprague-Dawley(SD)rats were used in the study.The original generation cells were obtained by modified tissue piece inoculation.The passaged cells were used.The ASMC were divided into 2 groups:H2S group of different dosages and control group.The contents of sodium hydrosulfide in the culture media of H2S group were 10-5,10-4 and 10-3 mol/L respectively,and the same volume of normal saline was added to control group.Each group was treated for 24 hours.Morphology of the cells was observed by inverted microscope and identified by immunohistochemical method employing ?-smooth muscle-actin.The expressions of SUR2B and Kir6.1 were identified by immunohistochemical SP technology.The SUR2B mRNA and Kir6.1 mRNA levels were assayed by real time fluorescence relative quantitative polymerase chain reaction.Results The passaged cells developed typical growth pattern peak and valley and the 97th percent of the cells expressed the smooth muscle specific differentiation marker ?-smooth muscle-actin.SUR2B and Kir6.1 could be detected in ASMC by immunohistochemical technology.They were both located at cytoplasmic and cytomembrane but not in at nucleus.Compared with control group,SUR2B mRNA and Kir6.1 mRNA levels in H2S groups were higher than those in control group in a dosage-dependent mode.Conclusions H2S can increased the expression KATP channel SUR2B mRNA and Kir6.1 mRNA levels of ASMC.J Appl Clin Pediatr,2009,24(1):21-23

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China ; Databases, Genetic ; Enterovirus ; genetics ; isolation & purification ; Evolution, Molecular ; Humans ; Molecular Epidemiology ; Sequence Analysis ; Sewage ; virology

Objective To study the influence of the optimized pre-hospital emergency transfer sys-tem on the safety, success rate and prognosis of pediatric patients. Methods A clinical data analysis was made of 1 015 cases of critically ill children delivered with the not-optimized pre-hospital emergency transfer system as group Ⅰ( Jan. 2007-Dec. 2009 ) and those of 1 431 with the optimized pre-hospital emergency transfer system as group Ⅱ(Jun. 2010-Dec. 2012). The preparation time for dispatch,critical illness scores before and after transport,transfer success rates and outcomes were compared between the two groups. Re-sults Before transfer,the critical illness score for group Ⅰ was 81. 73 ± 18. 11,for group Ⅱ78. 45 ± 20. 96, with groupⅡ being more critical(t=4. 154,P<0. 001) and needing respirator supporting and more vasoac-tive agents(50. 5%vs. 30. 2%;60. 8% vs. 51. 6%;P<0. 01). The dispatch delay for groupⅠand groupⅡwere(19. 34 ± 6. 45)min and (16. 19 ± 5. 89)min,respectively,actual time out for groupⅡwas shorter than that for groupⅠ(P<0. 001). The critical illness scores on arrival of ward were 83. 01 ± 16. 73 in groupⅠ, 83. 97 ± 17. 50 in group Ⅱ,the score for group Ⅱ being more improved than that for group Ⅰ( P<0. 05 ) . The transfer success rates, cure and improvement rates were higher in group Ⅱ compared with group Ⅰ(97. 6% vs. 94. 8%;94. 5% vs. 91. 0%;P<0. 01). Conclusion The application of the optimized pre-hos-pital emergency care system can improve the transfer efficiency and success rate,safety and treatment effect, reduce the mortality rate of pediatric patients.

BACKGROUND:The intracel ular accumulation of reactive oxygen species leads to oxidative stress. Hypoxia is widespread in physiological and pathological condition. Variation of bone proliferation and differentiation when bone tissues cultured or bone cel s induced toxicity by reactive oxygen species under hypoxia have not yet been reported. OBJECTIVE:To observe the biological characteristics of MC3T3-E1 pretreated with different concentrations of hydrogen peroxide (H2O2) in hypoxia, thus understanding the cel mechanism underlying prolonged bone healing in the elderly with osteoporosis and diabetes. METHODS:The MC3T3-E1 cel s pretreated with different concentrations of H2O2 were cultured in different oxygen concentrations. The proliferation of MC3T3-E1 was detected by cel counting kit-8. The cel differentiation was detected through alkaline phosphatase staining and alizarin red staining. Total RNAs were extracted and used for analyzing the mRNA levels of col age type 1, alkaline phosphatase and Cbfa1. RESULTS AND CONCLUSION:When MC3T3-E1 pretreated with 200μmol/L H2O2 for 6 hours, the cel proliferation was increased with time, but lower than that in the control group. The alkaline phosphatase activity was weakened, and the number of mineralized nodes was decreased at the early stage of differentiation. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours, the cel proliferation was decreased obviously. The alkaline phosphatase activity was stil weakened, and the number of mineralized nodes was decreased further, but not affected by hypoxia. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours and then cultured in hypoxia, the mRNA expression of Cbfa1 was decreased, but the mRNA expressions of col age type 1 and alkaline phosphatase were significantly increased. These results suggest that MC3T3-E1 pretreated with low concentration of H2O2 show a significant decrease in proliferation, while MC3T3-E1 pretreated with a high concentration of H2O2 and cultured in hypoxia show a decrease in osteogenic differentiation, especial y at the early stage of alkaline phosphatase formation.

Objective To investigate the impact of optimal respiratory system dynamic compliance (Cdyn) strategy for titrating positive end-expiratory pressure (PEEP)on the prognosis of acute respiratory distress syndrome (ARDS) in infants.Methods A total of 30 patients with ARDS admitted in Pediatric Intensive Care Unit (PICU) of Guangxi Maternity and Child Health Hospital were divided into 2 groups randomly (n =15).PEEP was set in the control group according to PEEP/FiO2 in American ARDS collaboration while optimal Cdyn was for the treatment group.Pressure control ventilation(PCV) mode and small tidal volume(7 mL/kg)strategy were taken.Respiratory mechanics,hemodynamics and inflammatory cytokines were monitored in each group before and after experiment.The time of assisted ventilation,and the hospital stay in PICU and 28-day mortality were compared.Results (1) There was no significant differences in gender,age and severity of disease between the 2 groups (all P ＞ 0.05).(2) The optimal PEEP of the control group was significant lower than that of the treatment group[(6.4 ± 1.4) cmH2O vs (9.9-± 1.6) cmH2O,P ＜ 0.01].(3) Cdyn and oxygenation index(OI) in 2 groups were all improved,and the degree of improvement in the treatment group was significantly better than that of the control group [Cdyn after the experiment at 2 h:(0.39 ± 0.03) mL/(cmH2O·kg) vs (0.36 ±0.03) mL/(cmH2O · kg),P＜0.05; 24 h:(0.40±0.03) mL/(cmH2O · kg) vs (0.38±0.03) mL/(cmH2O · kg),P ＜0.05; 48 h:(0.43 ±0.02) mL/(cmH2O · kg) vs (0.40 ± 0.02) mL/(cmH2 O · kg),P ＜ 0.01 ; OI after the experiment at 24 h:(20.07 ± 2.12) cmH2O/mmHg vs (21.94 ± 2.05),P ＜0.05 ; 48 h:(17.51 ± 1.64) cmH2O/mmHgvs (19.82 ± 2.07) cmH2O/mmHg,P ＜ 0.01] ; There were no significant differences in heart rate(HR),mean aerial pressure(MAP),cardiac index (CI) before and after the experiment (all P ＞ 0.05).(4) Interleukin-6 (IL-6) in two groups all decreased gradually,and the decrease in treatment group was more obvious than that of control group after the experiment at 24 h:(84.58-± 9.11) ng/L vs (93.18 ± 9.27) ng/L,P ＜ 0.05 ; 48 h:(76.67 ± 9.23) ng/L vs (90.10-± 9.42) ng/L,P ＜ 0.01.(5) The lengths of assisted ventilation and the hospital stay in PICU of the treatment group were significantly less than that of the control group [length of assisted ventilation:(6.62 ± 1.26) d vs (8.06-± 1.44) d ; the hospital stay in PICU:(8.12 ± 1.31) d vs (9.53 ±1.42) d,all P ＜0.05].There had no barotrauma and the mortality was not distinct between the two groups(P ＞ 0.05).Conclusions Optimal Cdyn method for titrating PEEP improves respiratory mechanics of ARDS obviously,shortens the time of assisted ventilation,and has no serious adverse effect on hemodynamics.

ObjectiveTo evaluate the effect of auscultation, partial pressure of carbon dioxide in end-expiration (PETCO2), transillumination technique to judge whether the endotracheal tube is misplaced into the esophagus. Methods A blinded randomized controlled trial was conducted. Sixty patients with American Society of Anesthesiology (ASA) gradeⅠ-Ⅱundergoing endotracheal intubation in Fengxian Central Hospital admitted from September 2014 to February 2015 were enrolled. Two endotracheal tubes with the same size were respectively inserted into the trachea and esophagus for the same depth after general anesthesia by the same person. Two blinded anesthetists with different experience checked the tube position using three methods including auscultation, PETCO2, and transillumination technique, respectively. The order of the tubes tested (trachea or esophagus) and the method used were randomized according to randomise numbers table. The experienced anesthetists conducted the test first, followed by an inexperienced anesthetist conducting the same methods. The numbers of right and wrong determinations with different methods by different anesthetists were recorded.Results Sixty patients underwent the procedures for 180 times, with intratracheal intubation for 90 times, and esophageal intubation for 90 times. It was shown that the results were not different in two groups [96.7% (174/180) vs. 92.2% (166/180),χ2 = 3.500,P = 0.057]. By using auscultation, the correct rate of experienced anesthetist was higher than that of inexperienced (95.0% vs. 78.3%,χ2 = 5.786,P = 0.013). Using PETCO2, both anesthetists were correct in all cases, and the accuracy was 100%. Using transillumination, the experienced anesthetist was mistaken in 3 cases (accuracy was 95.0%), while the inexperienced mistook in 1 case (accuracy was 98.3%), and no significant difference was found between two groups (χ2 = 0.500,P = 0.250). The correct rate of using transilluminaion was significantly higher than that of using auscultation (χ2 = 7.563,P = 0.004). The sensitivity and specificity of the auscultation was 70.0% and 80.0%, that of transillumination technique was 96.7% and 93.3%,and PETCO2 was 100%, respectively, for two groups.ConclusionsPETCO2 is the most reliable method for determining tube position, and it is superior to auscultation and transilluminaion. Transillumintaion technique is superior to auscultation, irrespective of anesthetists' experience, while the accuracy of auscultation showed an obvious relationship with the anesthetists' experience.

Cerebral Palsy ; physiopathology ; Child ; Child, Preschool ; Humans ; Infant ; Motor Activity ; Posture ; Walking