Objective To develop a sensitive and stable substrate for enzyme‐linked immunosorbent assay(ELISA)and carry out methodology evaluation tests on it .Methods A kind of substrate with tetramethyl benzidine and urea peroxide as its main compo‐nents were made up ,its sensitivity ,precision and storage stability were evaluated and compared with other three kinds of substrate . Results This substrate was more sensitive than two of the three commercial substrates .The variable coefficient of precision test was 3 .5% .The optical densities(OD) of the new substrate after stored one day ,seven days ,one month ,six months ,twelve months in 4 ℃ environment were 2 .268 、2 .403 、2 .358 、2 .278 、2 .330 respectively ,the standard deviation was 0 .056 185 ,the variable coeffi‐cient was 2 .4% .Conclusion The substrate proposed in this paper displays good sensitivity ,precision and stability .The preparation method is simple ,reproducible .This substrate not only satisfies the requirements of laboratory research ,but also meets with the de‐mands for commercial kit development due to its function as an important component of test kits .It is an economical and effective choice for both laboratories and research transformation .

Objective To explore the mechanism and therapeutic effects of wet dressing with 50％ magnesium sulfate ( MgSO4 ) as a part of comprehensive treatment for wound resulted from pit viper bites in children.Methods A total of 61 children with pit-viper-bitten wound enrolled from May 2009 through May 2011 were divided into two groups,namely the comprehensive treatment group (n =31 cases) and the conventional treatment group (n =30 eases). In the comprehensive treatment group,50％ magnesium sulfate wet dressing applied to the swelling parts of body in the early stage in addition to the conventional treatment and topically bandaged once a day keeping wet with spraying saline on the top of dressing.Before the treatment and on the 1st day,the 3rd day and the 5th day of treatment,the values of myocardial enzyme and the circumference of swelling parts were recorded,and the differences were analyzed by t- test.The differences in rate of wound healing in one week between two groups were analyzed by chi-square test.Results Before the treatment,there was no statistical difference in clinical features between the two groups with different modes of treatment for the viper-bitten children (P ＞ 0.05 ). During comprehensive treatment,the values of myocardial enzyme and the circumference of swelling parts decreased more markedly than those in conventional treatment group ( P ＜ 0.05 ).The rates of wound healing in one week of two groups were 80.5％ and 56.6％ respectively.Conclusions The comprehensive treatment with adjuvant of 50％ MgSO4 for the viper-bitten children can effectively reduce the tissue damage caused by venom,and the swelling as well,restoring function of body and shortening the time for wound healing and preventing complications.

Objective To explore how the severity of periventricular-intraventricular hemorrhage (PIVH)impact on physique and neurodevelopment in premature infants.Methods A total of 80 preterm infants with PIVH admitted to NICU of Qingdao Women and Children's Hospital from December 2013 to June 2015 were eligible.According to the Papile classification,the premature infants were divided into 4 groups.They were grade Ⅰ PIVH group,grade Ⅱ PIVH group,grade Ⅲ PIVH group and grade Ⅳ PIVH group.The infants with grade Ⅰ PIVH and grade Ⅱ PIVH belong to the low-grade PIVH group.The infants with grade Ⅲ PIVH and grade Ⅳ belong to the severe-grade PIVH group.All of them were regularly followed up for 12 months.Neurodevelopmental outcomes of infants at 6 and 12-month correction age were assessed by using the 20 items neuromotor assessment applying for 0-1 year old and the Bayley scales of infant development-Ⅱ.The differences in physical and neurophysical development of premature infants among 4 groups were compared.Results There were no significant differences in physical growth indicators such as body weight,body length and the incidence of weight growth retardation among 4 groups (all P＞0.05).The incidence of neurobehavioral abnormalities in infants with grade Ⅲ-Ⅳ PIVH was significantly higher than that of infants with grade Ⅰ-Ⅱ PIVH at 12-month correction age (21.05％ vs 3.28％,x2 =4.284,P=0.038).Physical development index(PDI) of grade Ⅰ-Ⅱ PIVH infants was significantly higher than that of grade Ⅲ-Ⅳ PIVH infants at 6-month correction age(F=11.500,P＜0.05).At 12-month correction age,grade Ⅰ-Ⅱ PIVH infants showed a significant higher mental development index(MDI) scores and PDI scores than those of grade Ⅲ-Ⅳ PIVH infants(F=14.227,16.515,all P＜0.05).Of the 80 cases assessed,infants with grade Ⅲ-Ⅳ PIVH had significantly higher rates of cerebral palsy(21.05％ vs 1.64％,x2 =6.300,P=0.012) and developmental delay (26.32％ vs 4.92％,x2=5.185,P=0.023) compared with grade Ⅰ-Ⅱ PIVH infants.Conclusions The severe PIVH can have negative effect on the neurodevelopmental outcomes of preterm infants and might induce mental retardation,cerebral palsy and other neurodevelopmental disabilities.Therefore,the regular follow-up and early intervention in preterm infants with PIVH should be implemented to improve the quality of their lives.

Objective To learn the current anesthesiologists deployment at the hospital,and to figure out how to measure anesthesiologists deployment.Methods Existing data and expert consultation were used to obtain various indicators,and the current anesthesiologists deployment was analyzed,calculating with the formula the needed number of anesthesiologists.Results Despite the rising demand and resource utilization,anesthesiologists were found in obvious shortage at the hospital.The number of anesthesiologists needed was calculated as 101.4,with a vacancy of 11.4.Conclusion The national health authorities were recommended to revise the standard of anesthesiologists deployment at hospitals.

Objective To investigate pulmonary function of tidal respiration in late premature infants with non-mechanical ventilation treatment at corrected gestational age of full term. Methods A total of 26 late premature infants with non-mechanical ventilation treatment at corrected gestational age of full term were collected from Qingdao Women and Children′s Hospital between April and December 2013,and saved as the late premature infant group. Also, a total of 31 full term infants with hyperbilirubinemia or neonatal ABO hemolytic disease were recruited and treated as the full term group. Then the parameters of pulmonary func-tion of tidal respiration were compared between the two groups. The late premature infant group was equally allocated into extrauterine growth retardation and non-extrauterine growth retardation groups according to the increase in weight and the ratio of time to peak tidal expiratory flow and total expiratory time(TPEF/TE) and the ratio of expiratory volume at peak tidal expiratory flow and total expiratory volume( VPEF/VE) were respectively compared in these two groups. Another correlation analysis between body weight and TPEF/TE was carried out in the non-extrauterine growth retardation group. Results (1)Gestational age difference had no significant significance between late premature infant group and full term group ( P <0. 05 ) . The body weight[(2. 39 ± 0. 44)kg vs. (3. 21 ± 0. 43) kg] and height[(46. 51 ± 3. 22) cm vs. (50. 16 ± 2. 14) cm] in late premature infant group were still lag behind that in full term group and the difference had significant significance(P<0. 05,respectively). (2)The comparison of parameters of pulmonary function of tidal respi-ration:the TPEF/TE [ ( 41. 74 ± 10. 94 )% vs. ( 48. 17 ± 11. 79 )%] , VPEF/VE [ ( 42. 66 ± 9. 66 )% vs. (48. 31 ± 9. 94)%],VPEF[(8. 02 ± 2. 85) ml vs. (10. 23 ± 3. 56) ml] and minute ventilation[(0. 76 ± 0. 23) L/min vs. (1. 00 ± 0. 44) L/min] in late premature infant group were significantly lower than those in full term group(P<0. 05,respectively). (3)The TPEF/TE and VPEF/VE all had significant significance in both late premature infant group and full term group(late premature infant group:r=0. 958,P=0. 000; full term group:r=0. 950,P=0. 000). (4)The TPEF/TE and VPEF/VE in non-extrauterine growth retardation group were closer to those in full term group. The TPEF/TE and VPEF/VE in extrauterine growth retardation group were lower than those in non-extrauterine growth retardation group(t= -2. 775,-2. 109,P=0. 011, 0. 047). Conclusion Airway(especially small airway) resistance of late preterm infant group is higher than that of term infants. Extrauterine growth retardation influences the airway(especially small airway) resist-ance.

To discuss bacteria of urine infection reoccurrence , the routine culture and L-form bacteria culture of urine tract infection on 850 cases were adopted and clinica l watching and analysis of the bacteria plasmid were held ．Result: of the l28 cases, re-infec tion rate l5.0%．,the positive rate of the routine bacteria culture for those initial diagnosis patie nts was 58.3% while that of the re-infection patients was 34.0% P<0.0l ．The initia l diagnosis L-form inspected rate 5.6%yet that of re-infection was up to 32.0 % P<0.01 ．The analysis of the pla smid bacteria of re- infection were of the same origins ．Conclusion: the first infection dont nee d L-form culture but re- infection should have L-form culture ．Analysis: plasmid positive bacteria of r e-infection was the result of the first infection not completely cured．

Objective To establish a simple,accurate and practi cal method of extracting plasmid from gram-negative and gram-positive bacteria ,Method we selected,Biswajit sahas method and comported others method, such as controlling pH andtemperature strictly and increasing EDTA-Na2 capacity． Result The eight strip s of Ecliv 5l7 can not be found completely in original method, but can be found in our one．We found the largest plasmid molecular weight was l86.3?kb, and t he minimum was l.l?kb from each 30 strains STA, EC, PA． Plasmid profile is alike through repeating ex p eriment three times． Conclusion:The method can be widely used in extracting bac teria plasmid and fit gram-negative and gram-positive bacteria whenever molecular weight ma x or min． These conditions can be obtained in general laboratory．

Objective To learn the suitability and validity of readmission service at counties and townships in rural China.Methods Seven pilot counties were randomly selected from the eastern,middle and western areas of China,and five diseases of the respiratory infection and cerebral system were set as the targets,while 600 target inpatient readmitted who had any of the five target diseases from 2012 to 2013 in such counties were selected from the NRCMS database.With expert consultation and empirical evaluation,the fitness of the days of stay at township hospital and necessity of hospitalization at county hospitals were evaluated for such target inpatients.Results 64.7％ of the target inpatients were found with unfit days of stay at township hospitals,mostly too short;59.0％ of the target inpatients' hospitalization at county hospitals were found with serious risk factors during their hospitalization;18.5％ of the inpatients were found with unnecessary hospitalization.Conclusions Hierarchical medical system can enhance the efficiency of the heahhcare system,but the current validity of readmission is poor,so is the fitness of days of stay at township hospitals,and the quality of the referral pattern.These problems mostly result from poor medical competence at township hospitals and high autonomy of patients in seeking medical service anywhere in the country.

Moyamoya disease is a rare cerebrovascular disorder, characterised by progressive stenosis and/or occlusion of the intracranial internal carotid artery and its proximal branches with the development of a basal collateral network. Moyamoya disease has a high prevalence in Asia, particularly in Japan, Korea and China. Ischemic events and intracranial bleeding are the most common clinical manifestation of moyamoya disease. Although the benefi cial effect on hemorrhage is still not clear, revascularisation surgery remains the most effective way to prevent the progression of ischemic symptoms. Moyamoya disease has been investigated by numerous studies since it was fi rst described 50 years ago, many conundrums remain to be solved. In this article, we review the history, epidemiology, aetiology, clinical manifestation, diagnosis and treatment of moyamoya disease. Recent advances and future challenges of moyamoya disease are also discussed.

AIM: To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells. METHODS: The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR. RESULTS: A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established. CONCLUSION: The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).