The major goal of microbial ecology is to study the structure and function of complex micro-bial communities. New molecular biological techniques have been successfully applied to analyze mi-crobial community structure. However they do not provide information on the physiologic properties of the detected microorganisms. A new tool for structure-function analyses in microbial ecology, micro-autoradiography combined with fluorescence in situ hybridization (MAR-FISH) can be used to simul-taneously examine the phylogenetic identity and the specific activity of microorganisms within a com-plex microbial community at a single-cell level. This article reviews the principle, experimental steps of MAR-FISH technique. The application of this technique in study of the environmental microbial com-munity and function is also summarized.

Objective To analyze the mutations in nucleotide sequences of transfusion transmitted virus(TTV) in neonatal infection.Methods Neonatal serum TTV-DNA was detected by a nested PCR technique.Fifteen Chinese neonates with positive TTV-DNA were diagnosed as TTV infection.ORF1 sequences of TTV-DNA from these neonates were determined.Results Homology of Chinese TTV(C01-C15) and Japanese TTV(N22)isolated ranged from 87.1%-97.7% at nucleotide level,but there were point mutations in Chinese TTV,such as GG→TT in locus 112 and 113,TTATC→CCTAT in locus 236-240.Conclusions Chinese and Japanese TTV isolated had the same genotype.Some gene mutations may increase the TTV pathogen,and result in neonatal hepatitis syndrome or hyperbilirubinemia.

Inheirted metabolic disodrers(IMD)involves in multiple substance dysbolism,which usually results in irreversible neurological lesions because of various categories and complicated clinical manifestations.In resent years,IMD became one of the hot spots in medical domain around the world,original diagnostic technique and management progressed unceasingly.This paper provides an overview of the traditio-nal detection and treatment about IMD,and reviews the new techniques such as gene analysis,gene chip,organ transplantation and enzyme replacement therapy at the same time.

Objective To compare the difference of early trauma experience between early-onset and late-onset obsessive-compulsive disorder(OCD),and the kinds of early trauma in the cause of OCD. Methods Three hundred and twenty-six patients with obsessive-compulsive disorder and 480 healthy con-trols were enrolled. Early Trauma Inventory-Short Form ( ETI-SF) was applied to assess the early trauma ex-perience,compare the difference of early trauma experience between early-onset ( n=138) and late-onset ( n=188) OCD,and the kinds of early trauma in the cause of OCD. Results There were significant differences in the early trauma experience between obsessive-compulsive disorder group and control group((3.90±3.61) vs (1.88±2.61), P<0.01).Scores of physical((1.12±1.38) vs (0.71±1.23), P=0.001),emotional((1.58± 1.71) vs (0.42±1.01), P<0.01),and sexual abuse((0.27±0.59) vs (0.09±0.36), P=0.001),and a gener-al traumatic experience((0.94±1.17) vs (0.66±1.09), P<0.01) were significant difference between patients and controls. Except sexual abuse((0.30±0.66) vs (0.24±0.67), P=0.42),there were significant differ-ences in the early trauma between early-onset and late-onset OCD((5.12±3.58) vs (3.01±3.38), P<0.01), especially emotional abuse((2.18±1.76) vs (1.12±1.53), P<0.01).247(75.8%) OCD patients and 79(57. 9%) healthy controls experienced early trauma(χ2=21.48, P<0.01). Experiencing one kind of early trau-ma,the prevalence of OCD was 39.1%,two kind was 61.0%,three kind of early trauma was 65.8%,and ex-periencing four or more kind of early trauma,the prevalence of OCD was up to 84.4%. Conclusion OCD patients have much more early trauma,especially early-onset OCD. Experiencing more kind of early trauma, and the prevalence of OCD higher may associated with the development of obsessive-compulsive disorder. It is important to consider the role of childhood trauma in the prevention and treatment of OCD.

Objective To highlight the characteristics of pulmonary sequestration (PS) with combination of practical experience and review of literature. Methods One patient with PS confirmed by biopsy was described and the relevant literatures of 279 cases were reviewed. Results PS can be divided into two types: intralobar type and extralobar type. The percentage of intralobar type is about 78.5%, intralobar type is 13.3%, other unreported is 8.2%. Clinically, PS was often combined by repeated pulmonary infection. The mis-diagnose rate is 72.8%. Etrograde arteriography or enhanced CT scan could find the abnormal artery, from which we can make correct diagnosis. The diagnose rate is 100%. Conclusion For patients with above clinical and X-ray symptoms, PS should be considered. Enhanced CT or retropgrade arteriography examination should be done as soon as possible. Timely diagnosis and surgery treatment are important for curing this kind of disease.

Objective To investigate the effect of MDR1 and MDR3 gene silence by shRNA of human breast carcinoma cell line MCF-7/Adr,and explore the role of MDR1 and MDR3 in adriamycin-resistance of breast carcinoma cells. Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 gene was transfected into cells. The control group was transfected with empty vector. The concentration of adriamycin was detected by the flow cytometry (FCM). Cell apoptosis was analysed by FITC-Annexin-V/PI double staining. Cell viability and the IC50 of adriamycin on MCF-7/Adr cells were determined by MTT method. MDR1 and MDR3 mRNA were assessed by RT-PCR. P-gp expression was detectedby immunochemistry. Results After treatment with ABCB1 and ABCB4 shRNA plasmid vector, the apoptosis of MCF-7/Adr cells was (30.21±1.65)%and (22.07±2.17)% respectively. Compared with untransfecedgroup and empty vector transfection group the difference was significant(P＜0.01). MDR1 and MDR3 shRNAcould increase cellular adriamycin accumulation of MCF-7/Adr cells. MCF-7/Adr cells viability and the IC50were significantly decreased after transfection. Compared with untransfeced group and empty vector transfectiongroup, the mRNA level of MDR1 and MDR3 in MCF-7/Adr cells were decreased by (89.5±0.8)%and(85.1±1.2)%, the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner. Immunochemistry proved that the expression of p-gp was significantly inhibited. Compared with untransfeced group and empty vector transfection group the difference was significant (P＜0.05). Conclusion The shRNA can effectively and specifically silence the expression of MDR1 and MDR3 gene, reverse the adriamycin-resistance mediated by P-gp in MCF-7/Adr cells. The reversal effect of adriamycin-resistance by shRNA of MDR1 is more effective than that of MDR3.

Objective To test intraoperative radiotherapy with mobile photon beam using the INTRABEAM system ( Germany) , and to analyze the dosimetric characteristics of low?energy photon beam using X?ray source and spherical applicators and explore its potential limitations in clinical application. Methods A special water phantom, a parallel?plate ionization chamber, and an electrometer were used to measure the depth dose rates and isotropy of dose distribution in x/y plane of X?ray source and different spherical applicators in the INTRABEAM system. Those data were then compared with the system data. Results For the X?ray source, the deviation of observed depth dose rate and isotropy in the x/y plane from the system data were-2.16%± 1. 36% and-1.9%~ 2. 1%, respectively. For applicators with different diameters, the deviation of observed depth dose rate, transfer coefficient, and isotropy in x/y plane from the system data were-10.0%~2. 3%,-8.9%~4. 2%, and-1.6%~2. 6%, respectively. Surface dose rate and dose gradient became larger with the decrease in the diameter of the spherical applicator. The measurement of depth dose rate and isotropy of X?ray source and spherical applicators showed good repeatability. The influencing factors for measurement accuracy included the positioning error of ionization chamber, energy response, noise current, and correction factor f ’ ( R ) . Conclusions This study reveals the dosimetric characteristics of the INTRABEAM system, verifies the accuracy of the system data, and obtains the data for clinical application and routine quality assurance. However, large dose gradient and small therapeutic range may limit its wide clinical application.

Objective To explore the application value of two-dimensional echocardiography with spatio-temporal im-age correlation (STIC) in fetal congenital heart disease (CHD) prenatal ultrasonography. Methods 11 036 hearts of fetus were inspected by severalviews order scanning method and for STIC volume database acquisition and off-line a-nalysis congenital heart disease fetus for autopsy after induced labor or contrast the follow-up results after birth. Re-sults 97 cases with spatio-temp-oral image correlation in 176 cases congenital heart and great vessels exception (dysrhythmias not including) with simple two-dimensional echocardiography,92 cases were accordant (one case with incorporative intracardiac malformation missed diagnosis);in screened congenital heart disease fetus,STIC (n=79) and routine ultrasonography (n=87) took (7.76±2.42) min and (9.68±2.13) min per case,respectively;in dif-ferent gestational weeks,the quality of the images derived from volume datasets were comparable to that directly ob-tained from 2D echocardiography. Conclusion STIC technology can be used as effective supplementary means of 2D echocardiography, and the combination can further improve the prenatal diagnosis of fetal congenital heart disease.

Objective:This work aims to explore the long-term efficacy and complications of late-course accelerated hyperfrac-tionation (LCAHF) for treating nasopharyngeal carcinoma. Methods:A total of 58 patients who consulted from December 2005 to May 2008 and histologically proven nasopharyngeal carcinoma at initial diagnosis were randomized into an LCAHF group (experimental group) and a conventional fractionation (CF) group (control group). The treatment dose for both groups was 2 Gy per fraction once dai-ly, 5 days a week. After the 40 Gy to 50 Gy dose, the dosage in the LCAHF group was increased to two daily doses at 1.5 Gy per frac-tion 6 h apart, 5 days a week. The total dose in this group was 73 Gy to 76 Gy, the total dose in the CF group was 70 Gy to 76 Gy, with the total course of the treatment shortened by 0.5 weeks to 1.5 weeks in the former group. Results:The 5-year control rates of the naso-pharyngeal cancers was 86% in the LCAHF group and 59% in the CF group (P=0.021), with statistically significant differences be-tween the two groups. The late complications slightly increased in the LCAHF group than in the CF group, but the differences were not statistically significant. Conclusion:LCAHF treatment improves the local control of nasopharyngeal carcinoma without increasing the incidence of long-term complications.