Objective:To investigate the application value of hepatic vein outflow tract reconstruction with ringed polytetrafluoroethylene vascular in right lobe living donor liver trans-plantation.Methods:The retrospective and descriptive study was conducted. The clinicopatho-logical data of 4 donors and 4 recipients undergoing right lobe living donor liver transplantation in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School and 17 donors and 17 recipients undergoing right lobe living donor liver transplantation in the First Affiliated Hospital with Nanjing Medical University from June 2015 to August 2018 were collected. Of 21 donors, there were 10 males and 11 females, aged from 35 to 57 years, with a median age of 46 years. The median body mass of 21 donors were 64 kg, with a range from 56 to 72 kg. Of 21 recipients, there were 16 males and 5 females, aged from 21 to 68 years, with a median age of 42 years. The median body mass of 21 recipients were 63 kg, with a range from 47 to 77 kg. Observation indicators: (1) surgical and postoperative situations; (2) follow-up. Follow-up was conducted by outpatient examination or telephone interview to detect graft function, tumor recurrence, vascular graft complications, patency of vascular graft and survival of recipients up to August 2020. All recipients will be followed up for all their lives. Measurement data with normal distribution were represented as Mean±SD and measurement data with skewed distribution were represented as M (range). Count data were represented as absolute numbers or percentages. The Kaplan-Meier method was used to calculate patency rates of hepatic vein outflow tract and survival rates to draw patency curve and survival curve. Results:(1) Surgical and postoperative situations: the operation time, the weight of donor graft, graft to recipient weight ratio and duration of hospital stay of 21 donors were (367±72)minutes, (557±68)g, 0.89%±0.16% and (10+2)days, respectively. No major complication requiring reoperation or intervention occurred in any of the 21 donors. One donor undergoing mild bile leakage preserved peritoneal drainage for one week. All 21 recipients underwent classic orthotopic liver transplantation successfully. The time of hepatic vein outflow tract reconstruction in donor graft, operation time and time of anhepatic phase of 21 recipients were (24±4)minutes, (326±66)minutes and (42±6)minutes, respectively. The number of reconstructed middle hepatic vein in hepatic segment 5 and 8 were 18 and 15, with the diameter of (6.1±1.3)mm and (7.2±1.2)mm, respectively. The number of reconstructed inferior right hepatic vein were 10, with the diameter of (6.3±1.3)mm. The postoperative treatment time at intensive care unit and duration of hospital stay of 21 recipients were (1.5±0.9)days and (22.6±6.7)days, respectively. Ten of 21 recipients underwent postoperative complications. Five recipients underwent graft dysfunction including the level of alanine aminotransferase and aspartate aminotransferase >1 000 IU/L and the level of bilirubin slightly increasing, combined with increased ascites. Enhanced computed tomography scan showed congestion in the right anterior of graft and thrombosis in the middle hepatic vein of hepatic segment 5 and segment 8. All 5 recipients undergoing graft dysfunction recovered with normal liver function and ascites decreasing after symptomatic treatment including liver protection therapy, anticoagulation and albumin infusion. Two recipients underwent inferior vena cava thrombosis and intractable pleural effusion one month after operation. Vena cava venography examination showed thrombosis in the graft vascular. Of the 2 recipients, one case with collateral circulation formation recovered undergoing balloon dilatation and stent placement combined with anticoagulation therapy of warfarin. The other one case recovered after anticoagulation therapy of warfarin. One recipient undergoing bile leakage and abdominal infection with klebsiella pneumoniae recovered after symptomatic treatment. Two recipients undergoing abdominal infection or pulmonary infection recovered after symptomatic treatment. There was no serious complication or death during perioperative period. (2) Follow-up: all 21 recipients were followed up for 10 to 57 months, with a median follow-up time of 38 months. During the follow-up, no recipient underwent graft dysfunction and 2 recipients had tumor recurrence at postoperative 6 months. Six of the 21 recipients died within 2 years after operation including 3 cases dying of tumor recurrence, 2 cases dying of acute hemorrhage and 1 case dying of liver failure. There was no death caused by vascular graft complica-tions. The postoperative 1, 3, 6-month, and 1-year and 2-year potency rates of hepatic vein outflow tract in 21 recipients were 88.4%, 88.4%, 82.4%, 68.0% and 42.1%, respectively. The 6-month, 1-year and 2-year overall survival rates in 21 recipients were 100%, 94.4%, 71.4%, respectively.Conclusion:Application of hepatic vein outflow tract reconstruction with ringed polytetrafluoroethylene vascular in right lobe living donor liver transplantation is safe and feasible.

AIM: To clone EBV-LMP2A gene, construct and identify the recombinant retroviral vector and stable cell strains expressing EBV LMP2A. METHODS: The full-length EBV LMP2A gene was generated by RT-PCR amplification from B95.8 cells which contain complement nucleotide sequence of EBV LMP2A gene. The gene was ligated to T-vector and sequenced to construct retroviral vector consisting with LMP2A. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pGEZ-LMP2A and two auxiliary viral vectors pHIT456 and pHIT60 by lipofectAMINE2000. Viral titration was performed according to the instructions of the manufacturer. To establish L929 cell line stable expressing LMP2A, L929 cells were infected with recombinant retrovirus three times and selected by Zeocine. The Zeocine-resistant clones (L929/LMP2A) were screened for LMP2A expression by RT-PCR and Western blot. RESULTS: The recombinant retrovirus vector carrying LMP2A gene was constructed successfully. Transfection yield a titer of 5×10~8 infectious particles/L. The infected L929 cells were selected by Zeocine. Results of RT-PCR and Western blot indicated that L929 transgenetic cells could stably express EBV-LMP2A. CONCLUSION: The L929 cell line stably expressing LMP2A provides suitability for extraction of the LMP2A protein and preparations of the vaccine for the therapy of EBV-associated diseases.

objective To investigate preoperative donor and recipient assessment,choice of surgical options in living donor liver transplantation(LDLT).Methods The clinical data of 95 patients who underwent LDLT from January 1995 to October 2007 in our center were retrospectively analyzed.Of all,92 recipients were benign end-stage liver disease patients (including 45 patients with Wilson disease),and 3 were malignant hepatic carcinbma patients.Results Thirty-one fight lobes without middle hepatic vein(MHV),3 right lobes with MHV,51 left lobes with MHV.and 10 left lobes or left lateral lobes without MHV were obtained.All the donors recovered after operation. Recipients with benign end-stage liver disease were followed up for 1 to 86 months,and the 1-,3-,5-year accumulative survival rates were 89%(82 cases),78%(71 cases)and 73%(67 cases),respectively. The 1-,3-,5-year survival rates of patients with Wilson disease were 92%(42 cases),89%(40 cases)and 76%(34 cases),respectively. For the 3 patients with malignant hepatic carcinoma,2 died and 1 was alive and well. The copper metabolism was back to normal in both donors and recipients. Conclusions Establishment of a system for the safety of donors is basic for LDLT. The key to raise the recipients' survival rates is to choose the optimal surgical approach. LDLT is effective in treating Wilson disease.

Primary liver cancer, especially hepatocellular carcinoma, poses a serious threat to the life and health of the Chinese people. Given the insidious onset of liver cancer, less than 30% of hepatocellular carcinoma patients are considered for radical treatment at the initial diagnosis. Systemic anti-tumor therapy plays an important role in the treatment of advanced hepatocellular carcinoma. Immunotherapy of hepatocellular carcinoma has developed rapidly, and an increasing number of immunotherapy drugs, which can better control the progress of hepatocellular carcinoma and prolong the survival of patients, have become first- and second-line treatment options. This article reviews briefly the progress of immunotherapy for hepatocellular carcinoma in recent years.

OBJECTIVETo sum up the clinical experience of liver transplantation.

METHODA retrospective study was made in 11 patients receiving living donor liver transplantation (LDLT)/and 14 patients having orthotopic liver transplantation (OLT), including one time operation of reduced size liver retransplantation and one time operation of cadaveric liver retransplantation.

RESULTSThe voluntary donors were a sister and 10 mothers of recipients. The location of graft included 3 patients of segment II, III, part of IV (not including intermediate hepatic veins), 6 patients of segment II, III, IV (including intermediate hepatic veins), and 2 patients of V, VI, VII, VIII (not including intermediate hepatic veins). The weight range of graft was 270 - 620 g. Twenty-four recipients achieved a long-term survival and retained normal liver function during the follow-up. Only 1 patient died from serious rejection on the 72nd day postoperatively. Ten patients with hepatitis B cirrhosis were treated with lamivudine and anti-HBVIg, and HBV-DNA in serum was negative during the follow-up for 4 approximately 21 months. Copperoxidase, ceruloplasmin and main indexes of liver function became normal in all patients with Wilson's Disease. Postoperative complications included abdominal hemorrhage (2 patients), acute respiratory distress syndrome (5), acute rejection (4), and acute renal function failure (2).

CONCLUSIONSThe wise solution to improve the result of liver transplantation and optimize liver resources is the "multimodal approach", by which all kinds of techniques for liver transplantation including CLT, LDLT and RSLT should well developed.

Adolescent ; Adult ; Child ; Female ; Humans ; Liver Transplantation ; adverse effects ; methods ; Living Donors ; Male ; Middle Aged ; Reoperation

OBJECTIVETo explore the value of the microsurgical technique in the reconstruction of hepatic artery.

METHODSFrom September 2000 to June 2001, we performed liver transplantation for 11 patients including living related liver transplantation (4) and 7 orthotopic liver transplantation (7). Arterial reconstruction was performed under an operating microscope.

RESULTSNo patients developed hepatic arterial thrombosis and serious complication, nor death for multiple organ failure.

CONCLUSIONMicrosurgical technique in reconstruction of the hepatic artery can improve surgical outcome, not only in orthotopic liver transplantation but also in living related liver transplantation.

Adolescent ; Adult ; Child ; Female ; Hepatic Artery ; surgery ; Humans ; Liver Transplantation ; Male ; Microsurgery ; Middle Aged ; Treatment Outcome

Pancreas arises from dorsal and ventral anlagens on an embryological basis,with a difference in histocytology.Immunohistochemical staining for an anti-pancreatic polypeptide can be performed to discriminate between the dorsal and ventral pancreas because of the difference in the amount of pancreatic polypeptide contained by the dorsal and ventral pancreas.Differences of survival in patients were approved due to the difference in histocytology between the dorsal and ventral pancreas,including the ability of local invasion,lymph node metastases,and nerve plexus invasion.

Objective:To investigate the clinical value of split domino donor auxiliary liver transplantation.Methods:The retrospective and descriptive study was conducted. The clinco-pathological data of 3 liver transplantation recipients who were admitted to Nanjing Drum Tower Hospital affiliated to Nanjing University Medical School and 1 liver transplantation recipient who was admitted to external hospital in September 2018 were collected. The first case was male, aged 22 years, who was diagnosed as type II citrullinemia (CTLN2). The second case undergoing liver transplantation in external hospital was male, aged 59 years, who was diagnosed as decompensated alcoholic cirrhosis. The third case was female, aged 52 years, who was diagnosed as hepatocellular carcinoma of right lobe of liver. The fourth case was female, aged 51 years, who was diagnosed as hepatocellular carcinoma of right lobe of liver. The donor liver from a brain and cardiac death donor was split in vitro into the left liver and the right liver, in which the right liver without middle hepatic vein, and the modified piggyback liver transplantation using the left liver and the classical orthotropic liver transplantation using the right liver was conducted on the first and the second case, respectively. The original liver of the first case was split in vivo into the left liver and the right liver, and the piggyback auxiliary liver transplantation using the left liver and the piggyback auxiliary liver transplantation using the right liver was conducted on the third and the fourth case who underwent extended right hemihepatectomy, respectively. Observation indicators: (1) intraoperative situations; (2) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect liver function, liver imaging, complication and survival of recipients up to October 2021.Results:(1) Intraoperative situations. Liver transplantation was conducted successfully on the first, third and fourth case, with the operation time, the volume of intraoperative blood loss, the donor liver cold ischemia time, the graft-to-recipient weight ratio were 400 minutes, 370 minutes, 390 minutes, 600 mL, 1 300 mL, 1 600 mL, 230 minutes, 152 minutes, 135 minutes, 1.2%, 0.8%, 1.1%. (2) Follow-up. B-ultrasound examination of the first, third and fourth case after liver transplantation showed that the blood flow was normal, and all the 3 cases discharged and were followed up at postoperative 1, 6 and 12 month. The liver function, the level of blood ammonia and citrulline were normal of the first, third and fourth case at postoperative 1 week. Imaging examina-tion showed normal liver morphology of the first and third case, and a transplanted liver atrophy caused by portal vein steal of the fourth case. ① The level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), direct bilirubin (DBil) of the first case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 22.8 U/L, 404.1 U/L, 355.5 U/L, 289.6 U/L, 31.0 U/L, 23.1 U/L, 42.1 U/L and 25.8 U/L, 31.5 U/L, 517.7 U/L, 327.6 U/L, 172.9 U/L, 15.9 U/L, 21.4 U/L, 47.5 U/L and 29.7 U/L, 3.8 μmol/L, 92.1 μmol/L, 87.4 μmol/L, 79.7 μmol/L, 90.1 μmol/L, 130.6 μmol/L, 33.8 μmol/L and 25.4 μmol/L, 2.3 μmol/L, 47.0 μmol/L, 44.1 μmol/L, 47.1 μmol/L, 57.4 μmol/L, 70.9 μmol/L, 24.7 μmol/L and 9.7 μmol/L, respectively. The level of citrulline and blood ammonia of the first case before and after liver transplantation were 999.0 μmol/L, 196.0 μmol/L and 14.6 μmol/L, 9.0 μmol/L, respectively. The first case was followed up for 3 years and survived without any liver transplantation related complication. ② The level of ALT, AST, TBil, DBil of the third case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 21.3 U/L, 143.9 U/L, 182.0 U/L, 132.0 U/L, 17.2 U/L, 10.1 U/L, 17.6 U/L and 16.8 U/L,20.0 U/L, 291.0 U/L, 227.5 U/L, 106.4 U/L, 15.8 U/L, 10.8 U/L, 17.1 U/L and 19.4 U/L, 6.8 μmol/L, 50.9 μmol/L, 45.0 μmol/L, 34.0 μmol/L, 32.4 μmol/L, 22.3 μmol/L, 12.8 μmol/L and 14.9 μmol/L, 2.5 μmol/L, 18.4 μmol/L, 17.2 μmol/L, 14.9 μmol/L, 14.8 μmol/L, 12.1 μmol/L, 3.6 μmol/L and 4.4 μmol/L. The level of citrulline and blood ammonia of the third case after liver transplantation were 24.9 μmol/L and 16.0 μmol/L. The third case was followed up for 3 years and survived without any liver transplantation related complication. ③ The level of ALT, AST, TBil, DBil of the fourth case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 35.0 U/L, 268.7 U/L, 682.0 U/L, 425.8 U/L, 57.5 U/L, 34.0 U/L, 29.4 U/L and 18.1 U/L, 37.0 U/L, 419.1 U/L, 436.2 U/L, 139.5 U/L, 35.2 U/L, 32.4 U/L, 54.7 U/L and 32.8 U/L, 7.1 μmol/L, 64.2 μmol/L, 41.4 μmol/L, 17.6 μmol/L, 34.2 μmol/L, 48.7 μmol/L, 14.1 μmol/L and 21.8 μmol/L, 2.8 μmol/L, 18.9 μmol/L, 16.1 μmol/L, 6.0 μmol/L, 14.6 μmol/L, 26.7 μmol/L, 3.9 μmol/L, 11.8 μmol/L. The level of citrulline and blood ammonia of the fourth case after liver transplantation were 8.4 μmol/L and 47.0 μmol/L. One week after surgery, the transplanted right liver of the fourth case occurred atrophy due to blood stealing from the right branch of the portal vein. B-ultrasound examination showed that the reflux of the hepatic artery and hepatic vein was unobstructed. Immunosuppressants were discontinued 3 months after operation on the fourth case and there was no complication such as rejection, bile leakage, biliary stricture, thrombosis and vascular stricture during follow-up. The fourth case died of lung metastasis 19 months after operation.Conclusion:Split domino donor auxiliary liver transplantation can be used for the treatment of metabolic liver disease and advanced hepatocellular carcinoma.

The caudate lobe of liver is anatomically divided into three parts: Spiegel portion, inferior vena cava portion and caudate process. The caudate lobe of the liver is located in the dorsal side of the liver, adjacent to the inferior vena cava, the three hepatic veins, and the left and right portal veins. The location of the caudate lobe depends on the location of anatomical landmarks and the location of staining, especially negative staining techniques. The left approach is suitable for Spiegel resection, and the right approach is suitable for paracentral resection of the inferior vena cava and caudate process. The dorsal approach and anterior approach combined with other approaches can achieve complete caudate resection. This article showed the combination of multimodal approach with total caudate lobectomy, partial caudate lobectomy and laparoscopic caudate lobectomy.

Objective: To evaluate the effect of human CCR1 (hCCR1) gene overexpression on the migration of human bone marrow-derived mesenchymal stem cells (hMSCs) towards hepatocellular carcinoma (HCC), and to examine the application prospects of MSCs as gene delivery vectors in the treatment of HCC. Methods: The hCCR1 gene was subcloned into a lentiviral vector to generate the recombinant plasmid pLV-hCCR1. The pLV-hCCR1 plasmid and two other packaging plasmids were co-transfected into 293T cells using calcium phosphate, and the virus-containing supernatant was collected. hMSCs were then infected with the recombinant lentivirus, and the expression of hCCR1 mRNA and protein was analyzed by RT-PCR and Western blot, respectively. The effect of CCR1 gene overexpression on the in vitro migration of hMSCs was examined using the Transwell migration assay. Orthotopic nude mice models of HCC were established using the MHCC-97H-GFP cell line, and the mice were divided into two groups (n = 8 per group). hMSCs were then intravenously injected via the tail vein into the tumor-bearing nude mice to examine the effect of hCCR1 overexpression on the in vivo migration of hMSCs towards HCC. Unpaired Student’s t-test was used for two-group comparisons, and one-way ANOVA was used for multi-group comparisons. Results: Restriction enzyme digestion and DNA sequencing demonstrated that the recombinant plasmid pLV-hCCR1 was constructed successfully. The LV-hCCR1 lentivirus packaged by 293T cells has high infection efficiency in hMSCs, and hCCR1 was overexpressed in hMSCs after LV-hCCR1 infection. Transwell migration assay showed that hCCR1-transfected hMSCs had significantly enhanced migration towards HCC cell line-derived condition medium (CM) compared with the control RFP-hMSCs [(134.8±15.7)/LPF vs (83.5±10.9)/LPF, t = 10.40, P < 0.01]. In vivo migration experiment also demonstrated that there was significantly higher number of hCCR1-hMSCs localized within the MHCC-97H-GFP xenografts than hMSCs-RFP on day 14 following intravenous injection of hMSCs in mice [(86.7±14.1)/HPF vs (54.5±9.6)/HPF, t = -7.32, P < 0.01]. Conclusion Overexpression of CCR1 gene can significantly enhance the migration capacity of hMSCs towards HCC cells in vitro and in vivo. This study provides evidence for potential clinical application of MSCs as more effective delivery vehicles in cancer gene therapy.