Biliary atresia (BA) is the most common cause of persistent jaundice in infant,it can rapidly advance to biliary cirrhosis and death.Surgical treatment in time can decrease mortality,so it is essential to be differentiated from other diseases as early as possible.Main diagnosis methods of BA include clinical feature of patients,liver biopsy,and high-frequen cy ultrasonography,MR cholangiopancreatography (MRCP),hepatobiliary scintigraphy.High-frequency ultrasonography is superior to other methods with high diagnostic specificity and sensitivity.The progresses of ultrasonography in diagnosis of BA were reviewed in this article.

In order to assess whether gene transfection could be mediated by ultrasound in association with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm(2) and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal transfection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm(2) for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63+/-1.07)% vs (5.57+/-0.56)%, P<0.05]. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm(2) for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
Cell Survival/genetics ; Green Fluorescent Proteins/genetics ; Hep G2 Cells ; Poloxalene/*pharmacology ; Transfection ; Ultrasonics

Objective To study indole alkaloids of Ervatamia hainanensis. Methods The compounds were separated and purified by column chromatography with silica gel and Sephadex LH-20. The structures were identified by IR, MS, and NMR. Results Seven alkaloids were identified as: hainanervatasine (Ⅰ), hainanervatacine (Ⅱ), vobasine (Ⅲ), coronaridine (Ⅳ), 3-hydroxyl coronaridine (Ⅴ), 3-(2-oxopropyl) coronaridine (Ⅵ), and tabernamine (Ⅶ), respectively. Conclusion Compounds Ⅰ and Ⅱ are new compounds. Compounds Ⅳ and Ⅵ are isolated from this plant for the first time.

Objective To investigate whether ultrasound-mediated microbubble destruction(UMD) could enhance cationic liposome (CL) induced plasmid DNA delivery or not,and optimize the transfection conditions.Methods Multiple parameters were explored to obtain the optimal transgene efficiency by means of with or without serum in culture medium,various CL or nano-liposomal bubble(NB) concentrations,different time point of ultrasonic irradiation.The transfection efficiency was assessed by fluorescence microscopy and flow cytometer,and cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay.Results The serum could protect the cells but show little impact on transfection efficiency induced by CL.CL and plasmid DNA at a weight ratio of 4 ∶ 1 exhibited high transfection efficiency of (17.71-± 0.79)％ and high cell viability of (91.28 ± 0.76) ％.CL combining with ultrasonic irradiation at the time point of 1 hour could increase the transfection efficiency to (24.85 ± 0.78)％ (P ＜0.01).Higher transfection rate (32.47 ± 4.01) ％ was obtained by adding NB at the concentration of 10 ％ (P ＜0.05).Conclusions UMD accompanied with CL could enhance gene delivery effectively,which would provide a new method for gene therapy.

Objective To summarize the sonographic characteristics of high frequency ultrasound and elastography of thyroid microcarcinoma(TMC),and to analyze the causes of misdiagnosis.Methods The preoperative ultrasonic data of 245 suspicious TMCs in 202 patients,as confirmed by operation pathology,were retrospectively analyzed.Results Preoperative ultrasonography accurately diagnosed 221 TMCs,the diagnosis rate was 90.2％,and the misdiagnosis rate was 9.8％.Among the missed diagnosed lesions,18 lesions were nodular goiter,others were 3 nodular goiter with focal papillary hyperplasia of follicular epithelium,2 nodular goiter with adenomatous hyperplasia,1 focal lymphocytic thyroiditis,respectively.The thyroid lesions≤0.5 cm in diameter were more easily to misdiagnose.According to the importance of the ultrasonographic features of TMC,the order were aspect ratio (A/T) ≥ 1,irregular-shape,microcalcifications,low or very-low echo.Based on the above corresponding characteristics and considering other features together,the diagnostic accuracy rate were 94.1 ％,93.9％,92.4％ and 90.5％,respectively.Less blood supply and ill-defined boundary were the secondary sonographic signs of TMC.The elastographic scores of TMC were most showed 4 to 5 points.Diagnosis of TMC relied on elastography alone is less effective,but when elastograph diagnosis based on high frequency ultrasound,the diagnostic accuracy is much higher,especially when there is no calcification in the lesions.Conclusions High-frequency ultrasound has a very important value in the diagnosis of TMC,while elastography has certain assistant value on the basis of high-frequency ultrasonic diagnosis.

The value of color Doppler flow imaging (CDFI) and intravenous contrast-enhanced ultrasound (CEUS) for assessing the transplanted liver and early diagnosing complications by examining hemodynamic changes was discussed. Seventy-five patients with orthotopic liver transplantation (OLT) underwent CDFI. The following parameters were measured: peak systolic velocity (PS), resistance index (RI) and Doppler perfusion index (DPI) of the hepatic artery (HA), time average velocity (TAV) of portal vein (PV) and velocity of hepatic vein (HV) in different stages postoperation. And 11 patients of them received CEUS. Thirty healthy subjects were enrolled as controls. The results showed that: (1) In 23 patients without obvious complications, TAV of PV within 15 days post-operation was significantly higher than in controls (P<0.05), PS and DPI of HA within 7 days postoperation were lower, but RI was higher than in controls (P<0.05); (2) When the hepatic artery thrombosis (HAT) occurred, PS and DPI of HA were obviously decreased, but TAV of PV significantly increased like a high saw-tooth wave; (3) While rejection occurred, both TAV of PV and PS of HA were decreased with the increase in RI of HA, and the triphasic wave of HV disappeared and displayed as saw-tooth wave; (4) The incidence of biliary complications in liver transplantation was increased when DPI was reduced; (5) Seven cases of hepatic carcinoma relapse after OLT demonstrated hyperecho in the arterial phase and hypoecho in the portal and later phase on CEUS; (6) In 2 cases of HA thrombus, there was no visualized enhancement in arterial phase of CEUS, but enhancement during the portal vein and parenchymal phase. It was concluded that the hemodynamic changes of PV, HA and HV in the transplanted liver are valuable for assessing the transplanted liver and early diagnosing complications on CDFI and CEUS.
Bile Ducts/pathology ; Contrast Media/*administration & dosage ; Hemodynamics ; Hepatic Artery/pathology ; Infusions, Intravenous/*methods ; Liver Transplantation/*adverse effects ; Liver Transplantation/*methods ; Liver Transplantation/ultrasonography ; Perfusion ; Postoperative Complications ; Ultrasonography/*methods ; Ultrasonography, Doppler/*methods

This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.

Objective To investigate the optimal ultrasound exposure parameter on H22 neoplasms of mice meditated by ultrasound exposure combined with self‐made nanobubbles ,and then observe their therapeutic effect combined with cisplatin and their possible mechanism of anti‐tumor . Methods Thirty mice engrafts models with subcutaneous H22 neoplasms were established and divided into 6 groups randomly ,which received ultrasound exposure at different intensity and exposure time . The contrast enhanced ultrasound imaging ( CEUS ) was performed in every group at the four time points of before treatment and at 0 h ,24 h ,72 h after treatment . To obtain the optimal ultrasound parameters ,the tumor inhibitory effect was assessed by enhanced intensity ( EI) and microvascular density ( MVD) . The H22 tumor were treated by ultrasound exposure nanobubbles combined with cisplatin to observe their tumor growth inhibition rate ,and the microvessels density and nuclear associated antigen Ki‐67 proliferation index were measured by immunohistochemical staining . Results There was a statistically difference in enhanced intensity (EI) between the experimental groups and control group ( P < 0 .05) . With the increasing of ultrasound intensity and exposure time ,the tumor inhibitory effect was more obvious ,with an increasing side reactions . Except the simple ultrasound group ,there was a statistical difference in tumor inhibition ,the mean MVD and the tumor cell proliferation index (KI‐67) between control group and the other ultrasound therapy groups ( P<0 .05) . The tumor inhibitory rate was the highest ( tumor inhibition rate 70 .0% ) and the mean MVD and KI‐67 expression were the lowest ( P <0 .05) in the combination group comparing with the others . Conclusions The ideal ultrasound exposure parameter of tumor inhibition showed that exposure intensity chose 1 W/cm2 and exposure time chose 1 min or 3 min intermittence . The ultrasound exposure self‐made nanobubbles combined with cisplatin could enhance the tumor inhibitory effect .Its mechanism may be related to the decrease of microvascular density ,the inbition of tumor cell proliferation and the increase of tumor cell necrosis .

Objective To evaluate the differential diagnostic value and superior of biliary atresia(BA)in the infants with cholestatic hepatopathy by high-frequency ultrasonography (HUS).Methods After 4 hours fasting,124 infants with cholestatic hepatopathy were scanned with high-frequency US.The data of hepatic size and parenchyma,gallbladder,triangular cord (TC) sign,bile duct,right hepatic artery (RHA)and portal vein (PV) were observed and measured.Meanwhile,the other data were collected,which included the clinical diagnosis,blood biochemical tests,the MRCP and dynamic duodenal liquid color check finding,the pathological results after liver puncture biopsy and so on.Results In 124 infants with cholestatic hepatopathy,BA was found in 61 infants and ruled out in 63.TC thickness,RHA diameter,and gallbladder length and width exhibited significant differences between the group with BA and the group non-BA(all P ＜0.001).The correctness for the diagnosis of BA was 90.3％ by the combination of TC sign and abnormal gallbladder morphology,and 83.1 ％ by stool color,81.5 ％ by γ-GT,47.5 ％ by MRCP,83.3 ％ by dynamic duodenal liquid color check,95.2％ by the pathology after liver puncture biopsy,respectively.Conclusions HUS is superior to other diagnostic methods in BA with higher accuracy rate,noninvasion,simplicity and economy.

E.coli single-stranded DNA-binding protein (SSB) plays an important role in replication, recombination and repair of DNA and is thus crucial for the survival of the bacteria.We described a high expression and efficient purification scheme and kinetic assay of interaction with its substrate, single-stranded DNA (ssDNA). A ssb gene (537 bp) for encoding SSB was obtained by PCR amplification from E.coli K-12 genome. The expression vector of the fusion protein SSB was constructed by attaching ssb gene to pQE30. SSB fusion protein was expressed in M15 E.coli strain induced by IPTG. SDS-PAGE analysis revealed that the expected protein with a molecular weight 20.6kDa was soluble and amounted to about 30% of the total bacterial protein. SSB protein was purified by immobilized metal (Ni2+) chelation affinity chromatography and the purity was about 90%. The resulting SSB protein was a correctly folded tetramer analyzed by gel filtration. It could bind ssDNA with equilibrium dissociation constant (KD) of 4.79×10-7 mol/L as determined by surface plasmon resonance.