Trying to provide ultrasonic image-aid measures for quantitative diagnosis and dynamic monitoring of liver fibrosis, we propose two scoring methods for liver fibrosis tissue in vivo , based on ultrasound radio frequency (RF) time series in this paper. Firstly, RF echo signals of human liver were recorded in this study. Then one of the recorded frame RF data was demodulated to be B model image. After that, a region of interest (ROI) in the B model image was selected. For each point in the ROI, its all frame data were acquired so that RF time series were formed. An SMR (size measure relationship) fractal dimension and six spectral features were extracted from RF time series in the ROI. With relative deviation and Fisher's discriminant ratio, seven features were weighted and summed so that the liver tissues' scores were obtained, Score-rd and Score-fisher, respectively. Area under ROC curve (AUC) and a support vector machine (SVM) were used to evaluate whether these scoring methods would be useful in distinguishing normal and cirrhosis tissues. Experimental results are shown as follows: Score-rd's AUC was 0.843, while Score-fisher was 0.816, SVM classification accuracies were both up to 87.5%. This proved that our proposed scoring methods were effective in distinguishing normal and cirrhosis tissues. Score-rd and Score-fisher have potential for clinical applications. They can also provide quantitative references for liver fibrosis diagnosis.
Area Under Curve ; Fractals ; Humans ; Liver Cirrhosis ; classification ; diagnostic imaging ; Radio Waves ; Support Vector Machine ; Ultrasonography

Objective To study the effect of lgG Fc gene on JEV DNA vaccine immunity. Methods Gene encoding IgG Fc was amplified by nested-RT-PCR technique from BALB/c murine spleen cells. JEV prME protein gene was obtained with restriction endonuclease BamH Ⅰ/EcoR Ⅰ from the eukaryotic recombinant named after pJME, which was constructed by us before. Recombinant, named after pJME/IgG Fc, with above two genes encoding JEV prME protein and BALB/c murine IgG Fc was constructed, and was tested by restriction enzymes analysis and DNA sequencing, then was transfected into China hamster ovary (CHO) cells by Lipo-fectAMINE 2000. Distribution and expression of the fusion proteins encoded by JEV prME protein and BALB/c murine IgG Fc genes in transfected CHO cells were detected by immunofluorescence and Western blot. The BALB/c micc were vaccinated with pJME/IgG Fc via intramuscular injection. Then the cytotoxic T lymphocyt (CTL) activity were assessed by lactic dehydrogenase (LDH) and the neutralizing antibody titer were assessed by 80% plaque reduction neutralization test. Results Molecular weights (2001 bp, 2730 bp) of the two in- serts released from pJME/IgG Fc with two group of restriction analysis associated with BamH 1/EcoR I and BamH Ⅰ/Not Ⅰ were correlated to the expected theoretic results respectively. It was estimated that molecular weight (Mr) of the fusion protein was 101 x 103. The expression of the above fusion protein was mainly distribu- ted in endochylema of transfected CHO cells,and not much in membrane of transfected CHO cells. CHO cells transfected with pJME/IgG Fc could express the fusion protein at the 32th cell passage. After immunization, the CTL activity and the neutralizing antibody titer in the pJMF/IgG Fc vaccinated group increased significantly compared with other vaccinated groups(P <0.05). Conclusion The recombinant pJME/IgG Fc was construc- ted and transfected into CHO cells successfully, and CHO cellular lines expressed fusion protein encoded by JEV prME protein and BALB/c murine lgG Fc genes stably were obtained. IgG Fc gene could reinforce the cellular immunity and humoral immunity of JEV DNA vaccine.

Objective Construction and expression of recombinant plasmid fusing encoding prME protein derived from Japanese encephalitis virus （JEV） and GM-CSF of BALB/c mouse. Methods GM-CSF gene was ampli/ied by nested-RT-PCR from BALB/c spleen cells. JEV prME protein gene was eluted by the digestion with restrict/on endonucleases BamH I/EcoR I from pJME piasmid. Genetic fusion of prME protein and GM-CSF are subcloned info pcDNA3.1 （＋） eukaryotic vector, and named as pJME/GM-CSF.The recombinant was confirmed by restriction enzymes digestion and DNA sequencing, and then transfected into China hamster ovary （CHO） cells by Lipofectamine2000. pJME/ GM-CSF expression in CHO cells was examined by Western blot. Results We observed 2001 bp and 2472 bp DNA fragments when pJME/GM-CSF was digested with BamHI/EcoRI and BamHI/NotI respectively as expected. The estimated molecular weight of the fusion protein was 85kD. Conclusion The recombinant pJME/GM-CSF was constructed and transfected into CHO cells successfully with pJME/GMCSF stably expressed.

Objective To analyze the clinical characteristics of twin reversed arterial perfusion sequence (TRAP),and investigate its prenatal evaluation and clinical management.Methods Karyotype results and ultrasound data of 25 TRAP cases were retrospectively reviewed,including estimated weight and umbilical blood flow of acardiac twin,cardiac function and middle cerebral artery peak systolic velocity of pump twin.Various managements and the outcomes were analyzed.Results (1) Karyotype of amniotie fluid were tested in 16 pump twins.Mosaicism was found in 1 case (46,XX[36]/46,XY [14]).(2) According to the ultrasound evaluation,large acardia accounted for 87.0％ (20/23) cases.Abundant blood perfusion (inter-twin difference of umbilical resistance index ≤0.20) was indicated in 86.4％ (19/22) cases.Decompensation of cardiac function was suggested in 66.7％ (10/15) pump twins.Fetal anemia of pump twin indicated by middle cerebral artery peak systolic velocity＞1.5 multiples of the median was diagnosed in 75.0％ (12/16) cases.(3) The acardiac twin with abundant blood perfusion was more likely to be a large acardia than those without [94.7％(18/19) vs 1/3,Fisher exact test,P=0.04]; More pump twin with large acardia tended to have cardiac decompensation than non-large acardia pump twins [83.3 ％ (10/12) vs 0/3,Fisher exact test,P=0.02].(4) Eleven patients chose to terminate their pregnancies after being diagnosed.In 14 cases who continue the pregnancies,the survival rate of pump twin was 64.3％ (9/14).In 3 cases of non-large acardia without cardiac decompensation of pump twin,the patients selected conservative observation resulting in 2 term deliveries and 1 termination of pregnancy due to for exacerbation.Among 11 cases with large acardia,which the pump twins were complicated by cardiac decompensation or anemia,five cases selected conservative observation.One ended in spontaneous abortion; three exacerbated (one termination and two cesarean section before term with living births) ; one was stable until delivery.Another 6 cases received bipolar cord coagulation,and successful interruptions of acardiac blood flow were achieved in 5 cases among which 4 pump twins survived.Conclusions Prenatal diagnosis,cardiac function and fetal anemia of pump twin,together with the growth and blood supply of acardia are important indexes for prenatal evaluation of TRAP,on which our prompt management should be based.

Objective To study the adjuvant effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) coding gene on cellular immunity induced by Japanese encephalitis (JE) virus DNA vaccine. Methods GM-CSF coding gene was amplified by nested-reverse transcriptase-polymerase chain reaction (RT-PCR) technique from BALB/c murine spleen cells. Recombinant plasmids pJME/GM-CSF and pGM-CSF were constructed by JE virus (JEV) prM-E protein with GM-CSF coding gene or GM-CSF coding gene only, respectively. The plasmids were transfected into China hamster ovary (CHO) cells by Lipofectamine 2000. The coding protein expressions and distributions were detected by immunofluorescence. The BALB/c mice were vaccinated with indicated immunogens with or without GM-CSF gene. The changes of T lymphocyte subsets in the spleen and levels of intracellular cytokines, such as interferon (IFN)-γ and interleukin (IL)-4 of splenic cells from mice immunized with different immunogens were evaluated by flow cytometry. The cytotoxicity T lymphocyte (CTL) activity was assessed by lactate dehydrogenase (LDH). The data were compared by one-factor analysis of variance and least significant difference. Results The constructed recombinant pGM-CSF and pJME/GM-CSF were confirmed by restrict enzyme digestion and DNA sequencing. The expressions of the above proteins were mainly in the cytoplasm and minor on cell membrane. The percentage of CD4+ T lymphocytes in pJME/GM-CSF vaccinated group was (33.90±0.79)%, which was significantly higher than that of in other groups (t values were 9. 818, 6. 804, 6.594, 10.061, 9.380, and 17.675, all P<0.05). The percentages of CD4+T lymphocytes in pJME +pGM-CSF (0) and pJME+pGM-CSF (-3) vaccinated groups were (29.83±0.61)% and (29.70±0.51)%, respectively, which were both higher than that in pJME+pGM-CSF (+3) vaccinated group of (27.69+0.50)% (t=3.466, t=3.255, both P<0.05). The percentages of CD8+ T cells in pJME/GM-CSF and pJME+pGM-CSF vaccinated groups were both higher than that in empty vector (pcDNA 3.1+) group and JE inactivated vaccine vaccinated group (t values were 3.811, 2.627, 10.537, and 3.811, all P<0.05). The CTL activity in pJME/GM-CSF vaccinated group was (51.48±0.10)%, which was higher than those in other groups (t values were 22.868, 13.823, 5.377, 32.287, 34.632, and 53.795, all P<0.05). The IFN-γ/IL-4 ratios in pJME/GM-CSF, pJME+pGM-CSF (0) and pJME + pGM-CSF (-3) vaccinated groups were (19.13±1.36), (12.32±0.82) and (7.05±0.43), respectively, which were higher than those in other groups (P<0.05). Conclusion GM-CSF coding gene could enhance the cellular immune response induced by Japanese encephalitis DNA vaccine.

Objective To summarize the key nursing points in patients with monochorionic twins undergoing selective feticide with bipolar electrocoagulation.Methods Selective feticide with bipolar electrocoagulation were performed in 68 monochorionic twins with one twin anomaly.And the perioperative care was performed on the patients.Results Operations were accomplished successfully in 68 cases.No complications,such as infections,premature rupture of membrane and so on,were found in all cases seven days after operation.Conclusions The nursing key points include preoperative psychological nursing by interpretation of the operations,monitoring fetal heart sounds during operations,and close observation of body temperature,uterine contractions and fetal heart rate in pregnant women after operations.Careful perioperative nursing is helpful for improvement of operative success and to ensure normal development of fetus.

[Objective] To summarize the perioperative nursing points during the amnioreduction by fast and negative pressure drainage.[Methods] Amniodrainage and associated nursing care were performed in 93 hydramnios cases of pregnant women from January 2006 to December 2010,and the nursing key points were summarized.[Results] Operations were performed successfully in 93 hydramnios cases of pregnant women.No complications occurred in 92 eases 3 d after operation.Bellyache and uterine contraction occurred in one case 2h after operation,which indicated placental abruption,two dead fetuses were got out by cesarean section.[Conclusions] The nursing key points included preoperative psychological nursing by interpretation of the operation,monitoring fetal heart sounds and close observation of contrac-tions in pregnant women.Careful perioperative nursing for patients with hydramnios is important to improve the success rate and reduce postoperative complications.

Objective This study aims to investigate the effect of rehabilitation skills training on suicide and relapse prevention of patients with depression.Methods Eighty patients were randomly divided into two groups.One group accepted depression rehabilitation skills training and the other group accepted general health education for 4 weeks.Both groups were followed up by 12 months,and the number of relapse and suicide and the score of Health-related quality of life made by Word Health Organization (WHOQOL-BREF) were recorded.Results The rate of relapse (10.0％ vs.42.5％) and hospitalization (5.0％ vs.20.0％) were lower in skills training group than in control group (P＜0.05).Rate of seeking help of suicide was higher in skills training group than in control group (25.5％ vs.7.5％) (P＜0.05).The suicide mortality was insignificantly different between two groups (0.0％ vs.2.5％) (P＞0.05).The scores of WHOQOL-BREF were significantly higher in skills training group than in control group in follow-up (P＜0.05).Conclusions Rehabilitation skills training program can not only reduce the rate of relapse and suicide but also improve the quality of life of patients with depression.

Objective To research the role of mitochondrial DNA mediate the cultured human retinal pigment epithelium (hRPE) cell apoptosis induced by blue light and the relationship with time.Methods Established the blue light damage model of cultured hRPE cells in vitro with light emitting diode (LED) blue light density of (4.0-±0.5)mW/cm2 adjusted by FL-1D blue light illumination meter,and the illumination time was set as 0,0.5,1,2,4,6,12 and 24 hours,then the cells were grouped according to the illumination time.Immunofluorescence were used to identify the cells;the expressions of caspase-3,cleaved caspase-3,caspase-9,cleaved caspase-9,bax and bcl-2 were detected with Western blot.Quantitative PCR was used to detect the copy number of mitochondrial DNA and PCR was used to detect mitochondrial DNA 4977bp common deletion.Results Immunofluorescence results showed that the RPE65 protein was expressed in the cytoplasm.The expressions of bax were upregulated after illumination for 1 hour,cleaved caspase-3 were upregulated after illumination for 2 hours,caspase-3,caspase-9,cleaved caspase-9 were upregulated after illumination for 4 hours,while the expression of bcl-2 was downregulated after illuminated for 2 hours,with significant differences compared to the normal control group (all at P＜0.05).The copy number of mitochondrial DNA in 0.5,1,2,4,6,12 and 24 hours groups was downregulated,with significant differences compared to the normal control group (all at P＜0.05).The expressions of 4977bp common deletion in 0.5,1,2,4,6,12 and 24 hours groups were increased,with significant differences compared with the normal control group (all at P＜0.05).Conclusions Blue light can cause cell apoptosis,especially mitochondrial apoptosis,in hRPE probably motivated by mitochondrial DNA damage.

Objective:To investigate the feasibility and efficacy of transcystic injection of indocyanine green during laparoscopic cholecystectomy (LC) surgery in the treatment of patients with gallbladder stones combined with liver cirrhosis.Methods:The clinical data of 96 patients with cirrhosis who underwent LC for gallbladder stones with cholecystitis attacks in the Department of Hepatobiliary and Pancreatic Surgery of the Second People′s Hospital of Changzhou City from January 2018 to May 2022 were retrospectively analyzed. All patients were diagnosed by clinical history and auxiliary examination before surgery and underwent cholecystectomy according to their groups, which were divided into fluorescence group ( n=49) and white light group ( n=47) according to whether ICG was used or not, where the fluorescence group underwent LC in fluorescence mode after direct intraoperative injection of ICG via gallbladder. The two groups were compared in terms of identification time of the three tubes, operation time, intraoperative bleeding, intraoperative injury, intraoperative open abdomen and blood transfusion, postoperative drainage time, postoperative hospitalization time, postoperative complications and changes in infection and liver function indexes before and after surgery. The measurement data obeying normal distribution were expressed as mean±standard deviation( ± s), and independent sample t-test was used for comparison between groups. The measurement data obeying the skewed distribution were expressed by M( Q1, Q3), and the Mann-Whitney U test was used for comparison between groups. Count data were expressed as rates (%), and the chi-square test or Fisher′s exact probability method was used for comparison between groups. Results:The procedure was successfully performed in both groups, and the identification of triple-tube time, operative time, intraoperative bleeding, postoperative drainage time, postoperative hospital stay, postoperative ALT, postoperative GGT, and postoperative TBIL in the fluorescent group were (14.96±1.49) min, (52.14±7.36) min, 5(5, 10) mL, (1.61±0.61) d, (2.37±0.49) d, 31.5(22.0, 44.8) U/L, 38.0(21.0, 91.5) U/L, 18.0(11.5, 22.8) μmol/L, and (29.87±3.37) min, (84.36±13.25) min, 10(10, 20) mL, (2.70±0.69) d, (3.15±0.42) d, 45.0(28.0, 64.8) U/L, 73.0(32.0, 132.0) U/L, 23.0(16.1, 29.3) μmol/L in the white light group and the differences were statistically significant compared with the two groups( P<0.05). In the fluorescence group, there was no intraoperative injury and no cases of intraoperative opening, and there were 2 cases of postoperative complications, while in the white light group, there were 2 cases of intraoperative injury, 1 case of intraoperative opening, and 5 cases of postoperative complications, and there were no cases of blood transfusion in both groups. There was no statistically significant difference between the fluorescence group and the white light group when comparing the preoperative laboratory indexes of both groups ( P>0.05). When comparing the first postoperative white blood cell count, C-reactive protein, aspartate aminotransferase, and alkaline phosphatase indexes in the fluorescence group and the white light group, there was no statistically significant difference between the two groups ( P>0.05). Conclusion:When LC is performed in patients with symptomatic gallbladder stones combined with cirrhosis, intraoperative injection of indocyanine green via the gallbladder to visualize the gallbladder and bile duct structures is simple and easy to perform, and the safe and efficient dissection of extrahepatic bile ducts and gallbladder bed speeds up the procedure while reducing postoperative trauma.