Objective To explore the application value of tissue dispersion quantitative analysis in rat liver fibrosis stages and provide a reference for clinical non-invasive diagnosis of liver fibrosis.Methods Seventy-two male Wistar rats were randomly divided into experimental group (n =64) and control group (n =8).The experimental groups of rats were gavaged with the volume fraction of 60％ CCl4 olive oil solution to form different stages of liver fibrosis,the control group did wvith normal saline,All the rat underwent tissue dispersion quantitative analysis to obtain 12 elastic parameters,the differences in above parameters were compared among rats with different liver fibrosis stages,then the correlation with pathological stages were analyzed.Results Except for COMP,ASM,CORR,there were significant differences in all parameters among rats of different liver fibrosis (P ＜0.05),which were correlated with pathological stages(P ＜0.05).Among these parameters,％ AREA had the highest correlation coefficient (r =0.891,P =0.001).The ROC curve was made by ％AREA to estimate the fibrosis stage.the area under ROC curve for ％ AREA was 0.914 (≥ the control group + S0),0.963 (≥early liver fibrosis S1 + S2),0.969 (≥middle liver fibrosis S3),0.948 (early cirrhosis S4),respectively.Conclusions The technique of tissue dispersion quantitative analysis has a good evaluation value for liver fibrosis stages,it is able to provide a preliminary reference for clinical non-invasive diagnosis of liver fibrosis.

Objective To study whether there was significant difference between pregnant women , data and the results of prenatal screening of single intrauterine fetal death ( sIUFD) when twin pregnancy and singleton pregnancy for guiding the clinical prenatal screening and risk consulting .Methods By comparative study, 56 cases of sIUFD when twin pregnancy were recorded from 2011 to 2014 in Ningbo Prenatal Diagnosis Center , all were natural pregnancy , the sistens gestational weeks were less than 14 weeks , and 4 993 natural singleton pregnancy .The pregnant women , data and the results of serological prenatal screening between sIUFD and singleton pregnancy were analyzed by t-test and rank sum test .Separately , the 56 cases of prenatal screening , risk value was calculated according to the twins and singleton , then the difference were analyzed combined with the results of follow-up.Results Pregnant women , data of two groups were analyzed, there were no statistically significant difference between sIUFD and singleton pregnancy .The age of sIUFD and singleton was (27 ±3)year-old and (27 ±3)year-old respectively, t=2.56, P>0.05; the weight of sIUFD and singleton was (55.2 ±10.23 ) kg and (56 ±10.34) kg, t=4.268, P>0.05.The gestational weeks of sIUFD and singleton were (39.21 ±0.78)weeks and (39.1 ±0.91) weeks, t=1.3, P>0.05;the weight of newborn was (3.38 ±0.41) kg and (3.31 ±0.43) kg, t=1.9, P>0.05.The AFP multiple of median (AFPMOM) of sIUFD and singleton was 1.41(0.99,1.83) and 1.02(0.84,1.24), Z=5.337, P<0.05.Free human chorionic gonadotropin beta multiple of median of sIUFD was 1.04(0.62, 1.64) and 0.99(0.67,1.51), Z=0.275, P>0.05;unconjugated estriol multiple of median of sIUFD and singleton was 1(0.79,1.16) and 1.01(0.85,1.21), Z=1.334, P>0.05.Trisomy 21 risk of sIUFD and singleton was 7 750(2 200,28 000) and 5 300(2 000,12 000), Z=2.093, P<0.05, that had significant difference.The 56 cases of prenatal screening risk value was calculated according to the twins and singleton , among whom 42 cases had the same conclusion , 14 cases had the different conclusion .Among them, according to singleton calculation , 3 cases for high risk, according to the twin calculation of high risk for 17 cases,χ2 =12.1, P <0.05.According to follow-up, all newborns were normal.Conclusions For the natural pregnancy , sIUFD when twin pregnancy , if the sistens gestational weeks less than 14 weeks, the risk of prenatal screening results calculated according to singleton will be more reasonable , as for the prenatal screening for twin pregnancy , the method needs further exploration .

Objective To investigate the value of prenatal ultrasound in diagnosis and treatment of twin reverse arterial perfusion (TRAP) syndrome. Methods A retrospective study was performed in 5 TRAP cases, including ultrasound images, clinical data and pregnancy outcomes. The sonographic characteristics were summarized. Results Five TRAP cases were diagnosed during 13 to 28 weeks' gestation and confirmed after birth. Color Doppler unltrasonography revealed retrograde umbilical artery perfusion towards acardiac twin. Two of 5 cases ended up in induced abortion, 1 in spontaneous abortion, 1 was delivered at 37 weeks' gestation after ultrasound-guided feticide of the acardiac twin and 1 was monitored closely with ultrasound and delivered alive at 32~(+4) weeks' gestation. Conclusion Prenatal ultrasonography has great applicative value for TRAP syndrome in early diagnosis, choosing optimal treatment and prognosis assessment.

One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like HIN1, NS from classical swine H1NI, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.

Objective To investigate the effects of growth-discordant twin pregnancies on neonatal thyroid stimulating hormone (TSH) level and congenital hypothyroidism (CH).Methods A total of 3 444 live-birth twin neonates born between January 1,2012 and December 30,2014 in Ningbo City were enrolled.Blood samples via heel puncture were collected and tested.Incidence of CH in singleton and twin neonates was compared.Deviation of birth weight larger than 25％ in twin neonates was set as the criteria for discordant growth.TSH and 17 α-hydroxylase levels in CH twins and normal twins,with or without discordant growth,were compared.Chi-square and non-parametric statistics were performed for data analysis.Results The incidence of CH in twin neonates was 0.56％ (19/3 444),higher than that in singleton neonates [0.09％ (203/225 712),x2=76.225,P＜0.01].Among nineteen CH twins,CH occurred in both twins in eight cases (four twins) and in one of the twins in eleven cases.The gestational age at birth in the eight CH twins were less than 37 weeks,with four males and four females;five were low birth weight infants;one twin were dichorionic,and three twins were monochorionic.In the eleven cases of CH occurring in one of the twins,the gestational age was less than 37 weeks in nine cases,eight were low birth weight infants,six were male and five female;seven were monochorionic and four were dichoronic twins.Five cases of temporary hypothyroidism were all low birth weight infants among the growth-discordant twins.CH cases in growth-discordant group had lower birth weight than their normal twins [M(P25-P75),2 100 (1 800-2 600) vs 2 770 (2 530-2 960) g,Z=4.369],and a higher TSH level [15.4 (11.8-18.5) vs 6.4 (4.8-7.9) mU/L,Z=6.339] (both P＜0.05).In normal twins with or without discordant growth,the neonates with a lower birth weight had a higher TSH level [3.6(2.5-4.7) vs 2.4(1.8-2.9) mU/L,Z=0.962] in weight consistent group,compared with 6.0(4.4-7.8) vs 3.4(1.9-4.1) mU/L in weight inconsistent group (Z=4.369),both P＜0.05.Conclusions In the growth-discordant twins,neonates with a lower birth weight have a higher TSH level and a higher risk of temporary hypothyroidism.

Objective To disscuss and analyze the clinical effects of whole body mild hypothermia on MODS caused by cerebral hemorrhage.Methods One hundred and eighty patients with MODS caused by cerebral hemorrhage during the period of hospitalized from January 2013 to December 2015 in ICU of our hospital were divided into two groups (observation group and control group) randomly,90 cases patients in each group.Patients in control group were treated with conventional treatment intervention,patients in control group were treated with whole body mild hypothermia on the base of conventional treatment intervention,the score of MODS,the ratio of,VO2 and DO2,the time during the period of hospitalized in ICU,the time of mechanical ventilation,epilepsy incidence and mortality,NIHSS score and PADL score of the patients in these two group after treatment were compared.Results After the comparion,the score of MODS and the state of oxygen consumption and oxygen supply of the patients in observation group were better than the patients in control group,there was significant differences,and had statistical significance (P＜0.05);the time during the period of hospitalized in ICU and the time of mechanical ventilation of the patients in observation group were shorter than the patients in control group,there was significant difference,and had statistical significance (P＜0.05);the mortality were reduce significantly of the patients in observation group than the patients in control group,there was significant difference,and had statistical significance (P＜0.05);the epilepsy incidence of the patients in observation group and control has no significant difference,has no statistical significance (P＞0.05);the NIHSS score and PADL score of the patients in observation group were higher than the patients in control group,there was significant difference,and has statistical significance (P＜0.05).Conclusion The clinical effects of whole body mild hypothermia on MODS caused by cerebral hemorrhage have important significance,can effectively improve the balance of multiple organ dysfunction syndrome patients,can delay the progression of multiple organ dysfunction syndrome patients,improve the prognosis,it is worthy of clinical application and promotion.

Objective To analysis and summary the chromosome abnormal existing in old pregnant women from 2002 to 2013,and to provide basis for clinicians intervene the fetus with chromosome disorders.Methods The 4 539 pregnant women in Ningbo city from 2002 August to 2013 October accepted the fetal karyo type detection,were retrospective analyzed,the frequency of abnormal chromosomal karyotypes was calculated according to different age groups,and the pregnancy outcomes of the old pregnant women were followed up.Thechi-square testswere performed on the frequency dateof the abnormal chromosome karyotype,polymorphism,and serum screening of high risk for fetal chromosome detection of less than 35-years-old pregnant women.Results The total of advanced maternal age pregnancyduring the past 11 years in Ningbo City is 32 080,and the follow-up rate was 99.90％,there are 10 infants borned with chromosomal abnormalities,the 1 290 caseswere detected withadverse pregnancy.A total of 4 539 advanced maternal age pregnancyaccepted amniocentesis,in those we found 107 cases of chromosome abnormality fetus,116 cases of polymorphism.A total of 5 232 high-risk pregnant women accepted the serum screening in the same period (less than 35 years old),finding 135 cases of fetal chromosome abnormal and 69 cases of polymorphism.Conclusion To strengthen the prenatal diagnosis,especially for puerperae above the age of 39,will lower the birth rate of infants with chromosome disease and will be conducive to the high quality of population in Ningbo.

Objective To investigate rational management for pregnant women with intermediate risk of Down syndrome (DS) through a retrospective analysis of second trimester maternal screening results for DS.Methods Second-trimester triple maternal serum screening for DS was performed.High risk ( ≥ 1/270) pregnant women received prenatal diagnosis.Intermediate (1/1000 to 1/270) and low (＜1/1000) risk pregnant women received routine obstetric examination.Data of ultrasound diagnosis,karyotype analysis of amniotic fluid and postpartum follow-up were collected.Outcomes of pregnant women were retrospectively analyzed.The incidences of DS among three groups were compared.Data of different groups were compared with Fisher exact or Chi-square test.Results Eighty-six thousand eight hundred and seventy-four pregnant women accepted maternal serum screening and outcomes of 86 126 cases were followed up,among which 4342 cases were DS high risk,8196 intermediate and 73 588 low risk.The incidence of DS was 6.22‰(27/4342) for high risk,0.73 ‰ (6/8196) for intermediate risk and 0.04 ‰ (3/73 588) for low risk group.The incidence of DS was higher in high risk group than in intermediate risk group (Fisher exact probability,P=0.000),and was higher in intermediate risk group than in low risk group (Fisher exact probability,P=0.000).Comparison among the three subgroups within intermediate risk group showed no significant differences (x2 =0.047,P =0.977).Conclusions Pregnant women with intermediate risk of DS should be paid more attention.Reasonable management such as ultrasound diagnosis and prenatal consultation should be provided.

To prepare the RNAse-resistant virus particles containing the partial gene fragments of avian influenza virus H5N1 for use as RNA standard and control in RNA virus detection, the genes coding the coat protein and maturase of E.coli bacteriophage MS2 were amplified by PCR and then cloned into prokaryotic expression vector pET32a to construct the intermediate vector pET32a-MS2. In addition, the gene sequences coding hemagglutinin (HA), neuraminidase(NA) and M protein of the H5N1 virus were also cloned separately to the down-stream of plasmid pET32a-MS2, thus constructing the prokaryotic expression vectors pET32a-NS2-HA, pET32a-MS2-NA and pET32a-MS2-M. These recombinant plasmids were then transformed separately to E.coli BL21(DE3) with induction by IPTG. to express the virus-like particles. The virus-like particles observed under electron microscopy were identified by RT-PCR ,while their stability was confirmed by real-time RT-PCR. In this way, the virus-like particles were successively constructed and identified through PCR amplification, enzymolysis identification and sequencing analysis. These virus-like particles observed under electron microscopy appeared to be circular in shape with a diameter of about 50 nm. Their stability was proved to be rather good. From these observations, it is apparent that these virus-like particles can be used as RNA standard and quality control in the detection of avian influenza virus H5N1.

Objective To prepare and study the immunogenicity of hepatitis B virus surface anti-gen (HBsAg)-tetanus toxoid (TT) conjugate vaccine. Methods Tr was activated by cyangen bromide and reacted with adipic acid dihydrazide, then HBsAg-TT conjugate was prepared by carbediimide. Conjugate, HBsAg or hepatitis B vaccine was injected subcutaneously into mice. Anti-HBsAg and HBsAg-specific T cell response elicited by these immunogens were assayed. Results New HBsAg-TT conjugate elicited higher levels of anti-HBsAg and HBsAg positive conversion rates after the immunization than did HBsAg alone or hepatitis B vaccine. Conjugate induced mesdy antibodies of the IgG2a subclass, while HBsAg alone or hepa-titis B vaccine mainly elicited anti-HBsAg in the IgG1 subclass. The number of IFN-γand IL-2 secreting T cells induced by conjugate was also significantly higher than that did by HBsAg or hepatitis B vaccine. Con-clusion This study indicated new HBsAg-TT conjugate can induce both stronger humoral and TH1 type of cellular immune response.