[Effects of FRNK on activation and migration of hepatic stellate cells].

Objective: To investigate the effect of focal adhesion kinase related non kinase (FRNK) on the activation and migration of hepatic stellate cells (HSCs). Methods: Human liver tissue was divided into healthy control group and fibrosis group from March 2019 to September 2019 in Affiliated Hospital of Guizhou Medical University. C57BL/6 mice were divided into wild type (WT) and FRNK gene knockout type (FRNK-/-) groups. The liver fibrosis model was established with carbon tetrachloride (CCl4). After that, FRNK gene overexpression (Ad-FRNK) was constructed with adenovirus vector. HE and Masson staining were used to evaluate the pathological changes and fiber deposition of liver tissue. Western blot was used to detect the expression of PY397-FAK and α-SMA protein. Mouse primary HSCs were extracted, and the effect of FRNK on HSCs migration was detected by wound healing, activation of Rac and Rho was detected by Western blot. Results: The expression of PY397-FAK protein in human liver tissue with hepatic fibrosis was significantly higher than that in healthy control group (0.88±0.09 vs. 0.73±0.09). FRNK was significantly lower than that in control group(0.68±0.09 vs. 0.79±0.11). After animal model was set up, the degree of liver fibrosis in FRNK-/-mice (153±13)% was more serious than that in WT (100%) group. The expression of PY397-FAK and α-SMA protein was significantly elevated (2.50±0.23 vs. 0.75±0.09, 1.46±0.20 vs. 0.92±0.10). After FRNK gene was re-expressed (100%), the degree of liver fibrosis was mainly reversed [(74±6)%], and the expression of PY397-FAK and α-SMA was accordingly decreased(0.68±0.11 vs. 1.12±0.19,0.68±0.10 vs. 0.85±0.06). In vitro, FRNK inhibited the migration of HSCs [WT∶FRNK-/-∶Ad-FRNK,(339±49)%∶(580±53)%∶(259±33)%] and the activation of Rac and Rho proteins (Rac: 0.54±0.07 vs. 0.91±0.10 vs. 0.77±0.12,Rho:0.45±0.05 vs. 0.64±0.06 vs. 0.53±0.07), all P<0.01. Conclusions: FRNK can inhibit the activation and migration of HSCs which contributed to liver fibrosis. The potential mechanism is related to down regulation of PY397-FAK and inhibition of Rac and Rho activation.

[Analysis of the pregnancy outcomes in pregnant women with COVID-19 in Hubei Province].

Objective: To study the effect of COVID-19 on pregnancy outcomes and neonatal prognosis in Hubei Province. Method: s A retrospective comparison of the pregnancy outcomes was done between 16 women with COVID-19 and 45 women without COVID-19. Also, the results of laboratory tests, imaging examinations, and the 2019 novel coronavirus (2019-nCoV) nucleic acid test were performed in 10 cases of neonatal delivered from women with COVID-19. Result: s (1) Of the 16 pregnant women with COVID-19, 15 cases were ordinary type and 1 case was severe type. No one has progressed to critical pneumonia.The delivery method of the two groups was cesarean section, and the gestational age were (38.7±1.4) and (37.9±1.6) weeks,there was no significant difference between the two groups (P>0.05). Also, there wee no significant differences in the intraoperative blood loss and birth weight of the newborn between the two groups (all P>0.05). (2) Ten cases of neonates delivered from pregnant women with COVID-19 were collected. The 2019-nCoV nucleic acid test were all negative.There were no significant differences in fetal distress, meconium-stained amniotic fluid, preterm birth, and neonatal asphyxia between the two groups (all P>0.05).(3) In the treatment of uterine contraction fatigue, carbetocin or carboprost tromethamine was used more in cesarean section for pregnant women with COVID-19 (1.3±0.6), compared with Non-COVID-19 group (0.5±0.7),the difference was statistically significant (P=0.001). Conclusions: If there is an indication for obstetric surgery or critical illness of COVID-19 in pregnant women, timely termination of pregnancy will not increase the risk of premature birth and asphyxia of the newborn, but it is beneficial to the treatment and rehabilitation of maternal pneumonia. Preventive use of long-acting uterotonic agents could reduce the incidence of postpartum hemorrhage during surgery. 2019-nCoV infection has not been found in neonates delivered from pregnant women with COVID-19.

Detection and analysis of laser driven proton beams by calibrated Gafchromic HD-V2 and MD-V3 radiochromic films.

The radiochromic film (RCF) is a high-dose, high-dynamic range dosimetry detection medium. A stack of RCFs can be used to detect both spatial and energetic distribution of laser driven ion beams with a large divergence angle and continuous energy spectrum. Two types of RCFs (HD-V2 and MD-V3, from Radiation Products Design, Inc.) have been calibrated using MeV energy protons and carbon ions produced by using a 2 × 6 MV tandem electrostatic accelerator. The proportional relationship is obtained between the optical density and the irradiation dose. For protons, the responses are consistent at all energies with a variation of about 15%. For carbon ions, the responses are energy related, which should be noted for heavy ion detection. Based on the calibration, the broad energy spectrum and charge distribution of laser accelerated proton beam with energy from 3 to 8 MeV and pC charge were detected and reconstructed at the Compact LAser Plasma Accelerator at Peking University.

Creation of Electron-Positron Pairs in Photon-Photon Collisions Driven by 10-PW Laser Pulses.

A novel approach is proposed to demonstrate the two-photon Breit-Wheeler process by using collimated and wide-bandwidth γ-ray pulses driven by 10-PW lasers. Theoretical calculations suggest that more than 3.2×10^{8} electron-positron pairs with a divergence angle of 7° can be created per shot, and the signal-to-noise ratio is higher than 10^{3}. The positron signal, which is roughly 100 times higher than the detection limit, can be measured by using the existing spectrometers. This approach, which could demonstrate the e^{-}e^{+} pair creation process from two photons, would provide important tests for two-photon physics and other fundamental physical theories.

High-efficiency γ-ray flash generation via multiple-laser scattering in ponderomotive potential well.

γ-ray flash generation in near-critical-density target irradiated by four symmetrical colliding laser pulses is numerically investigated. With peak intensities about 10^{23} W/cm^{2}, the laser pulses boost electron energy through direct laser acceleration, while pushing them inward with the ponderomotive force. After backscattering with counterpropagating laser, the accelerated electron is trapped in the electromagnetic standing waves or the ponderomotive potential well created by the coherent overlapping of the laser pulses, and emits γ-ray photons in a multiple-laser-scattering regime, where electrons act as a medium transferring energy from the laser to γ rays in the ponderomotive potential valley.

Dilemmas across different overseas liver transplant stages: Taiwan transplant recipient families' perspectives.

AIMS: This study aimed to explore the dilemmas of Taiwanese overseas liver transplant recipient families (OLTRF) across three overseas liver transplant (OLT) stages in Taiwan and Mainland China. PATIENTS AND METHODS: An exploratory qualitative method was employed using a purposive sample of OLTRF, who received guided face-to-face, semistructured interviews. Data were subjected to content analysis. RESULTS: Nineteen OLTRF (15 female, 4 male) aged between 29 and 71 years (mean 55.1) for 19 patients with end-stage liver diseases were interviewed. OLT stages including predeparture stage (first stage), stay in China stage (second stage), and reentry to Taiwan stage (third stage). Ten kinds of dilemmas were encountered: (1) unable to get transplantation immediately (first to second stages); (2) dilemma of choosing overseas transplantation (first to second stages); (3) uncertainty about the transplantation outcomes (second to third stages); (4) care pressure (second to third stages); (5) poor diet adaptation (second to third stages); (6) lack of trust in the medical care quality (second stage); (7) worry about not fulfilling family responsibilities (second stage); (8) lack of information (all stages); (9) financial pressure (all stages); and (10) frustration when seeking medical care (all stages). CONCLUSIONS: Taiwanese OLTRF's perspectives of their dilemmas through the OLT process were first revealed in this study. Both Western and Eastern health professionals might be empowered by better understanding of OLTRF's living experiences and concerns during the stages of overseas liver transplantation.

Motivations and decision-making dilemmas of overseas liver transplantation: Taiwan recipients' perspectives.

PURPOSE: This study sought to explore the motivation and dilemmas in the decision-making process encountered by Taiwan overseas orthotopic liver transplant (OLT) recipients. PATIENTS AND METHODS: We used an exploratory qualitative research method on a sample of transplant recipients. Face-to-face in-depth interviews were performed with a semistructured interview guide. Data were evaluated by qualitative content analysis. RESULTS: We interviewed 15 patients including 11 males and 4 females aged between 41 and 68 years (mean = 57.3) including 14 who received OLT in China and one in the United States. The most important motivations were saving lives from end-stage liver disease and avoiding a hopeless sense of their residual lives with the psychological torture of a waiting death. Their decision-making process leading to overseas OLT could be divided into several phases among which the phase of transplant data evaluation and decision was the most critical one. Nevertheless, every stage and phase had its unique contents, factors, and dilemmas. CONCLUSION: This study demonstrated that patients encountered various dilemmas at different phases in the decision-making process of considering overseas OLT. This information is important for care providers and policy makers in dealing with patients who consider overseas OLT.

Caring for overseas liver transplant recipients: Taiwan primary family caregivers' experiences in mainland China.

INTRODUCTION: This study explores the motivations of overseas liver transplant recipient (OLTR) families of Taiwanese OLTR who undergo the procedure in mainland China. We report the waiting time to receive the transplant in mainland China as well as the rational and service. PATIENTS AND METHODS: This exploratory qualitative method reflects guided face-to-face, semistructured interviews with families members of OLTRs. Data were subjected to content analysis. RESULTS: We interviewed 19 OLTR family members (15 females and 4 males who were between 29 and 71 years of age; mean 55.1 years) regarding 19 patients who had (17 males and 2 females who were between 36 and 71 years of age, mean, 54.6 years). The OLTR underwent transplantation in three cities in mainland China: Tianjin, Shanghai, and Guangzhou. After arrival the average waiting time was 33.1 days. Subjects reported the following reasons to help patients undergo the procedure in mainland China: (1) it is difficult to have the procedure in Taiwan; (2) the desire to extend life; and ((3)) there is no domestic living donor. Seven reasons for serving as OLTR supportive family members were identified: (1) The role and obligation in the marital relationship; (2) a close bloodline relationship; (3) insufficient manpower; (4) an individual's availability; (5) evasion of responsibility by other family members; (6) compensation for inadequate caring efforts earlier in life; and (7) an unwillingness to disturb other relatives' lives. Finally, the following support for the OLTR was reported: providing company during medical treatment/doctor visits, food preparation, massage, daily assistance, medical care, and psychological support. CONCLUSIONS: Taiwanese OLTR family members' perspectives throughout the transplant process may provide better understanding of living experiences and concerns during the stages of overseas liver transplantation.

Gene expression of P-glycoprotein and cytochrome P450 3A4 in peripheral blood mononuclear cells and correlation with expression in liver.

Long-term immunosuppression in renal transplant recipients generally includes calcineurin inhibitors (CNIs), which demonstrate marked interindividual diversity and a narrow therapeutic range. In the clinical setting, it is important to reach therapeutic drug concentrations to prevent allograft rejection. The same immunosuppressant dosage leads to different drug concentrations. Therefore, we investigated factors that influence the metabolism of immunosuppressant agents. The CNIs are substrates of cytochrome P450 (CYP450) and P-glycoprotein. The CYP450 3A genotype significantly influences CNI concentration. Differences in expression of these proteins may explain interindividual pharmacokinetic variations. However, it is risky and impractical to obtain specimens from the liver in renal transplant recipients. Therefore, we investigated the correlation of gene expression between peripheral blood mononuclear cells (PBMCs) and liver parenchyma. We observed that the correlation of relative P-glycoprotein gene expression between PBMCs and liver is not significant (r2=0.03; P=.65). In addition, the correlation of CYP450 3A4 gene expression between PBMCs and liver is not strong (r2=0.23; P=.42). The expression level of CYP450 3A5 is too low to be detected in the sample from PBMCs.

Frequent nuclear expression of beta-catenin protein but rare beta-catenin mutation in pulmonary sclerosing haemangioma.

BACKGROUND: Pulmonary sclerosing haemangioma (PSH) is an uncommon tumour that is composed of glandular/papillary lining cells and polygonal cells. The biological behaviour of this tumour has been investigated; however, the molecular pathogenesis of PSH remains unknown. AIMS: To characterise the role of the Wnt/beta-catenin pathway in the genesis of PSH. METHODS: 37 PSH samples were investigated immunohistochemically for detection of the beta-catenin protein and direct sequencing of exon 3 of the beta-catenin gene. RESULTS: Nuclear expression of beta-catenin was found in the lining component of 23 tumours (62%) and in the polygonal component of 11 tumours (30%). The expression of beta-catenin was stronger in the lining component, but weaker in the polygonal component. Interestingly, all the tumours with expression of beta-catenin in the polygonal component also expressed beta-catenin in the lining component. However, mutation of exon 3 of the beta-catenin gene was detected in only one tumour that expressed nuclear beta-catenin in lining and polygonal components. CONCLUSIONS: The Wnt/beta-catenin pathway is involved in the genesis of PSH, but mutation of exon 3 of the beta-catenin gene rarely contributes to the activation of the Wnt/beta-catenin pathway in PSH.

Effects of and predictors for tacrolimus rescue therapy among renal transplant patients under cyclosporine-based immunosuppression.

BACKGROUND: Though cyclosporine has dramatically decreased rejection rates and improved graft survival rates of renal allografts, there is still a remarkable rate of acute rejection and progressive deterioration of renal function after transplantation. Rescue therapy with tacrolimus has been used for allografts failing under cyclosporine-based treatment in order to get some renal functional recovery or stabilization. The aim was to evaluate tacrolimus rescue therapy for failing allografts under cyclosporine-based immunosuppression for possible prediction factors for success. PATIENTS AND METHODS: Thirty-five renal allograft recipients with failing transplants under cyclosporine-based immunosuppression were enrolled into this study. Renal function was evaluated by reciprocal serum creatinine level (1/Cr) and calculated CCr. The slope of changes in 1/Cr and CCr were calculated before and after tacrolimus therapy. The possible risk factors that affect the outcome of tacrolimus rescue therapy were analyzed. RESULTS: Nineteen patients showed improved renal function (group 1) and 16 patients, persistent deterioration (group 2) after rescue therapy. Group 1 showed positive slopes of changes of 1/Cr and CCr after rescue therapy. Group 2 patients showed persistent negative slopes although less negative than before rescue therapy. Only the posttransplant time was the significant predictive factor for successful tacrolimus therapy (P = .018). CONCLUSION: Tacrolimus rescue therapy improved or stabilized renal function in some patients with failing grafts under cyclosporine-based immunosuppression. To assure a successful rescue effect, it should be given early after transplantation, if there is a tendency toward deterioration of renal function.

Evaluation of cyclosporine C2 levels in long-term stable renal allograft recipients.

BACKGROUND: The use of cyclosporine was traditionally monitored by the trough level (C(0)). However, the immunosuppressive effects of cyclosporine correlate with its drug exposure, represented by the area under curve (AUC). It was also noted that cyclosporine C(0) level correlated with AUC poorly, while C(2) level (concentration at 2 hours after drug administration) satisfactorily correlated with AUC. Most recent studies concern the use of C(2) levels in de novo renal transplant patients; target levels of C(2) have been suggested. There is rare discussion about the C(2) target level for long-term cyclosporine-maintenance patients. Our objectives were to analyze the cyclosporine C(2) levels of patients more than 12 months after transplantation as well as changes in C(2) with time and the correlation between C(2) level and renal function. METHODS AND PATIENTS: This was a cross-sectional case-controlled study of 101 kidney recipients immunosuppressed with a cyclosporine-based regimen for at least 12 months. Both C(0) and C(2) levels were examined at various time points during outpatient clinic follow-up. The patients were stratified according to the time after transplant surgery, or to their renal function. RESULTS: The 101 patients were divided into three groups based on the time after renal transplant surgery. Groups 1, 2, and 3 represented patients transplanted for 1 to 3 years (n = 16), 4 to 6 years (n = 35), and more than 6 years (n = 50), respectively. The C(2) levels for each group were 657 +/- 232, 561 +/- 186, and 580 +/- 243 ng/mL, respectively, (P = NS). When stratified into low versus high C(2) groups, there were no significant differences in renal function both at the beginning and at the end of 1 year follow-up. Seven of 67 patients shifted to stronger immunosuppression in the low C(2) group, but only 2/34 in the high C(2) group, a difference that was not significant (P = .234 by Fisher Exact Test). Patients with creatinine levels greater than 1.5 mg/dL or lower than 1.5 mg/dL showed no difference in C(2) on C(0) levels. Patients with deterioration of renal function during this period had no different C(2) levels as those with no deterioration of renal function. CONCLUSION: The average C(2) levels among long-term cyclosporine-maintained patients were significantly lower than those previously suggested. C(2) levels did not correlate with the long-term outcome of renal function in patients at least 1 year after renal transplantation.

Use of simulect can reduce the incidence of acute rejection and demonstrates with superior 3-year patient and graft survival rates in renal transplantation.

BACKGROUND: Acute rejection is a major cause of graft loss in renal transplantation. Because the highest risk for acute rejection is in the first month posttransplantation, improved prophylaxis could be most beneficial in this period. Simulect administration provides 30 to 45 days of immunoprophylaxis against acute rejection during the critical period after transplantation. OBJECTIVES: We sought to assess the incidence of acute rejection episodes and the safety and tolerability of Simulect plus Neoral immunosuppression. Patient and graft survival rates up to 3 years posttransplantation were evaluated. METHOD: Forty-one transplant recipients received Simulect by intravenous infusion of an initial 20-mg dose on the day of renal transplantation and a second 20-mg dose on day 4 posttransplant. All renal recipients received immunosuppression with Neoral and steroid. RESULTS: There were eight cases (19.5%) of acute rejection within 1 year. The rejection episodes were easily reversed with steroid pulse therapy in seven patients except for graft loss. The 1-, 2-, and 3-year graft survival rates were 95%, 93%, and 88%, respectively. Overall, the 3-year patient survival rate was 100%. CONCLUSIONS: Simulect in combination with Neoral and steroid-reduced the incidence of acute rejection without an increase in adverse events. The low incidence and severity of acute rejection may have led to the superior 3-year patient and graft survival rates in renal transplantation.

Liver transplantation for acute hepatic failure.

INTRODUCTION: The mortality rate of acute hepatic failure (AHF) with conservative treatment is 40% to 90%, depending on the etiology. Hepatitis B infection is the major cause of AHF in Asia. In this study, we examined the role of liver transplantation for adult patients with AHF. METHODS: Sixteen patients with AHF received liver transplants in the past 6 years. Eight patients received cadaveric donor and another 8 living-related donor grafts. Fifteen patients suffered from hepatitis B-related disease and 1 had drug-induced AHF. Extracorporeal charcoal hemoperfusion was used as a bridge to liver transplantation in the first 2 patients and plasma exchange was used in the following patients. RESULTS: One patient died 1 month after the operation due to primary nonfunction. The other 15 patients are alive with good graft function at 2 months to 6 years follow-up. The success rate is 94%. Postoperative complications included infection in 10 patients (62.5%), acute rejection in 4 patients (25%), and biliary complication in 2 patients (12.5%). No neurological complications were noted. CONCLUSION: Liver transplantation is the most effective treatment for patients with AHF. Living donors may be considered due to the organ shortage and the critical patient disease.

Do patients with acute liver failure have a better chance to receive liver grafting?

OBJECTIVE: Patients with acute hepatic failure (AHF) were always given first priority on the transplant waiting list. We investigated whether AHF patients will deprive other patients on the waiting list of the chance of liver transplantation (LTx). METHODS AND RESULTS: From January 1999 to March 2003, a total of 423 patients were on the transplant waiting list at the National Taiwan University Hospital. Sixty-five of the patients had AHF caused by hepatitis-B-related disease (HBV, n = 52, 80%), Wilson disease (n = 3, 4.6%), drug-induced AHF (n = 3, 4.6%), and other causes (n = 7, 10.8%).Thirty-three patients died and 16 survived by medical treatment. Two received LTx abroad and 14 underwent LTx at our hospital (7 living-related; 7 cadaver). A total of 140 patients died while waiting for a transplant during the period studied. Of them, 107 were among 358 non-AHF patients (30%), and time-to-death interval was 133 +/- 175 days (median: 62); 33 were among 65 AHF patients (51%); time to death was 19 +/- 28 days (median: 8). There were 35 cadaver donor livers available during the period; 28 of 358 non-AHF patients (7.8%), and 7 of 65 AHF patients (10.7%) received cadaveric LTx. Their waiting time totaled 342 +/- 316 and 12 +/- 9 days, respectively (P < .0001). CONCLUSION: Most AHF patients died unless they received liver grafts. Even with a higher priority assigned to them, AHF patients still have little chance to get a cadaver donor liver in Taiwan, and non-AHF patients have an even slimmer chance. Therefore, we need to encourage liver donation from living-related donors.

Surgical complications and outcome of living related liver transplantation.

INTRODUCTION: Living donor liver transplantation (LDLT) is now widely performed for patients to resolve the critical shortage of organs from cadavers. Despite rapid implementation and expansion of the procedure, both outcome and complication analyses of LDLT are still incomplete. OBJECTIVES: To analyze the outcome of LDLT, with particular reference to complications of those in need of surgical or radiological intervention. METHODS: Forty-eight LDLTs performed at National Taiwan University Hospital between December 1997 and April 2003 were reviewed retrospectively. RESULTS: Forty-two (87.5%) patients survived the operation. The 1-year graft and patient survival rate was 81.5%. Seventeen of the 48 LDLT patients had at least one postoperative complication, which needed surgical or radiological intervention. The complications included bile leakage (n = 3), biliary stricture (n = 4), internal bleeding (n = 7), intra-abdominal abscess (n = 2), liver abscess (n = 1), hepatic artery thrombosis (n = 2), duodenal ulcer bleeding (n = 1), jejunal perforation (n = 1), adhesion ileus (n = 1), and intracranial hemorrhage (n = 1). Nine of the 17 patients with complications died. In contrast, only 2 of the other 31 patients died. Seven of the mortalities were related to the complications. All survivors received only one definite intervention early after the complications were diagnosed. However, the others received an average of 1.71 +/- 0.95 (0 to 3) interventions. CONCLUSIONS: Complications requiring surgical or radiological treatment caused major mortality of LDLT. Early and definite treatment of these complications is important to improve the patient's outcome.

Liver transplantation for patients with hepatocellular carcinoma.

BACKGROUND: Liver transplantation (LT) has been advocated as a salvage treatment for unresectable hepatocellular carcinoma (HCC). Selection criteria still need to be developed in Taiwan. OBJECTIVES: The purpose of our study was to assess the clinical findings and outcome of cirrhotic patients with HCC undergoing liver transplantation. METHODS: Our study consisted of 13 HCC patients who underwent liver transplantation during October 1996 to March 2003. The medical records and pathologic reports were analyzed retrospectively. RESULTS: Overall survival rates at 1 and 3 years were 86% and 61%, respectively. HCC recurrences occurred in three patients, one of whom is still alive with HCC recurrence 2 years after LT. The other two patients died of HCC recurrence 1 and 2 years after LT, respectively. Pretransplant alpha-fetoprotein (AFP) levels of >200 ng/mL were noted in all three patients with HCC recurrence. In contrast, only one of the ten patients without HCC recurrence had pretransplant AFP >200 ng/mL (P = .003). Four patients did not meet Milan criteria, two of whom had HCC recurrence. However, the other two patients with microscopic vascular invasion survived and were free of HCC. The only one patient, who had histologic grade 4 HCC, died of recurrence, although his tumor was AJCC stage 1. CONCLUSIONS: High AFP level is a risk factor for HCC recurrence after LT. In addition to Milan criteria, histologic tumor grading should be considered in patient selection. Microscopic vascular invasion may not affect the outcome of the patients with early HCC.