Objective To estimate an automatic DNA extracting method, and evaluate the quantity and quality of these DNA samples.Methods Genomic DNA was extracted automatically from blood samples by MiniPrep 75-Ⅱ workstation. The yield and purity of DNA samples were detected by UV-spectrophotometer, and the integrality of these DNA samples were measured by agarose gel electrophoresis.Results The yield of genomic DNA extracted from 100?l whole blood were (7.33?2.58)?g. The average purity of these DNA samples was 1.647?0.135 (A260/A280) and the molecular weight of them was about 21 kb.Conclusion High quality and intact genomic DNA can be extracted rapidly from whole blood by using MiniPrep 75-Ⅱ workstation, and these DNA samples were suitable for downstream molecular biology experiments.

Objective:To investigate the application of endoscopic ultrasound-guided liver biopsy (EUS-LB) to liver transplant recipients.Methods:In this retrospective cohort study, a total of 12 liver transplant recipients who underwent EUS-LB by the same endoscopist and specimens were diagnosed and reported by the same pathologist due to abnormal liver function or need to be evaluated for graft fibrosis in the Organ Transplantation Center of the Affiliated Hospital of Qingdao University were enrolled into the EUS-LB group from December 2021 to March 2022, meanwhile, a total of 23 patients whose PLB was completed by the same hepatologist and specimens were diagnosed by the same pathologist during the same period were enrolled in the PLB group. Acquisition of liver specimens and postoperative adverse events of the two groups were compared.Results:Patients in both groups were punctured 1-2 times on average, and the median total length of liver specimens in the EUS-LB group was significantly longer than that in the PLB group (61 mm VS 17 mm, Z=11.362, P=0.002). There was no significant difference in the length of the longest liver specimens between the two groups (17.6±6.9 mm VS 13.7±3.5 mm, t=2.382, P=0.086), while the number of liver specimens in the EUS-LB group was more than that in the PLB group (4.8±2.1 VS 2.3±1.2, t=9.271, P=0.001). The number of complete portal tracts was 11.3±4.6 in the EUS-LB group and 6.2±3.3 in the PLB group ( t=8.457, P=0.003). Abdominal pain was the only postoperative adverse event, and only 1 patient in the EUS-LB group had postoperative abdominal pain, which was fewer than that in the PLB group [8.3% (1/12) VS 43.5% (10/23), χ2=4.893, P=0.036]. Conclusion:Compared with PLB, EUS-LB delivers longer liver biopsy specimens with more complete portal tracts in liver transplant recipients, and fewer recipients complain about postoperative pain in EUS-LB group. Therefore, EUS-LB is a safer, more effective and more comfortable liver biopsy method.

Objective:To explore the clinical applications and therapeutic outcomes of immune checkpoint inhibitors(ICIs)on liver transplantation(LT)recipients after tumor development.Methods:Eight databases including PubMed, China National Knowledge Infrastructure, Wanfang Data, CQVIP, PubMed, EMBASE, Web of Science and Google scholar were accessed for searching the relevant literature articles in both Chinese and English from the establishment of databases to December 31, 2021. Disease response, adverse reactions and prognoses of patients with malignant tumors after LT and receiving ICIs were analyzed.Results:The patient was diagnosed as chronic rejection plus drug-induced liver injury by liver biopsy. After intermittent treatment with DPMAS plus plasma exchange and immunosuppressants, he finally died of tumor recurrence at 37 months after LT. After screening, a total of 28 articles on the application of ICIs after LT were retrieved. In these articles, there were 47 patients(37 males and 10 females)with a median age of 57(14-71)years and the predominant type of tumor after LT was hepatocellular carcinoma(28/47, 59.6%), followed by malignant melanoma in 11 cases (23.4%), non-small cell lung cancer in 3 cases(6.4%), colorectal cancer, cholangiocarcinoma, squamous cell carcinoma, hypopharyngeal squamous cell carcinoma and post transplant lymphoproliferative disease(PTLD) in 1 case(2.1%). The overall remission rate after ICI treatment was 29.8%(14/47)and the disease progression rate 68.1%(32/47). Among them, 31.9%(15/47)had immune rejection. Case fatality rate was 61.7%(29/47)and median survival time 6.5(0.3-48.0)months.Conclusions:Depending on existing publications, among those LT recipients with malignant tumors treated by ICIs, the rate of graft rejection and patient mortality are higher. ICIs should be carefully considered for LT patients and further researches are required.

Objective:To analyze the clinical characteristics of patients with de-novo non-alcoholic fatty liver disease (de-novo NAFLD) and patients with de-novo NAFLD combined with metabolic syndrome (MS) after liver transplantation (LT) , and to determine the related risk factors.Methods:Patients who underwent LT at the Organ Transplantation Center, the Affiliated Hospital of Qingdao University, from Jan. 2016 to Oc. 2020 and were monitored until Oct. 2021 were gathered. The recipients were divided into the group with/without de-novo NAFLD, and LT recipients with de-novo NAFLD were divided into the group with/without combined MS. Clinical characteristics of the LT recipients with de-novo NAFLD combined with MS were analyzed. Logistic regression analyses were performed to identify the risk factors for LT recipients with de-novo NAFLD and those with combined MS.Results:A total of 324 LT recipients with a median follow-up of 2.9 years (range: 2.0-4.3 years) were included in the study. De-novo NAFLD was diagnosed in 21.0% (68/324) of the LT recipients, and MS was diagnosed in 44.1% (30/68) of these patients. Compared with LT recipients without de-novo NAFLD, those with de-novo NAFLD had higher preoperative body mass index (BMI) , blood glucose, glycated hemoglobin levels and lower platelet levels, and longer postoperative follow-up, higher BMI, waist circumference, albumin, triglycerides (TG) , low-density lipoprotein (LDL) , blood glucose, glycated hemoglobin levels, and the incidence of MS (all P<0.05) . Preoperative platelets, glucose, postoperative albumin, LDL and BMI were independent risk factors for predicting de-novo NAFLD after LT (all P<0.05) . Preoperative glucose performed well in predicting the occurrence of de-novo NAFLD (threshold: 5.5mmol/L, P<0.001, AUC=0.678) . The differences in Pre-LT blood glucose, post-LT BMI, waist circumference, prevalence of prediabetes or diabetes, fatty liver index (FLI) , and NAFLD fibrosis score (NFS) between de-novo NAFLD LT recipients with and without combined MS were significantly different (all P<0.05) . Conclusions:The incidence of de-novo NAFLD after LT is noteworthy, and LT recipients with de-novo NAFLD are more likely to have a combination with MS. In preoperative treatment, keeping blood glucose to 5.5 mmol/L or below trends helps to lower the risk of de-novo NAFLD following LT. LT recipients’ nutritional state and lipid levels require prompt care. High albumin levels might not be a desirable thing. De-novo NAFLD LT recipients with concomitant prediabetes or diabetes may imply an increased risk of developing comorbid MS during the post-LT follow-up. Controlling FLI levels in LT recipients with de-novo NAFLD may reduce the risk of developing comorbid MS.

Objective:To investigate the clinical characteristics of de novo malignancies (DNMs) after liver transplantation (LT) and to study the clinical management strategies.Methods:Adult LT recipients who were regularly followed-up in the Organ Transplantation Center, the Affiliated Hospital of Qingdao University from January 2005 to April 2021 were enrolled in this study. The clinical characteristics of DNMs were retrospectively analyzed. Of 601 LT recipients, there were 105 females and 496 males, aged (51.4±9.6) years old. They were divided into the DNMs group ( n=26) and the non-DNMs group ( n=575) according to whether there were DNMs on followed-up. Clinical data including age, sex, basic diseases before LT and operation time were collected. These patients were follow-up in outpatient clinics. Results:Twenty-six patients were diagnosed to develop DNMs after LT, but there were 28 DNMs (of which 2 patients were diagnosed to have DNMs twice). The incidence of DNMs after LT was 4.3% (26/601), the median time from LT to DNMs was 42 (20, 70) months, and the cumulative incidence rates of DNMs were 0.5%, 2.0%, 6.3%, 21.0% and 34.5% at 1, 3, 5, 10 and 15 years after LT, respectively. Among the 28 DNMs, digestive system tumors were most common, with 17 lesions (60.7%), followed by 3 lesions (11.1%) of lung cancer, 2 lesions (7.4%) of lymphoproliferative diseases, and 1 lesion (3.7%) of cervical cancer, thyroid cancer, soft palate cancer, eyelid cancer, laryngeal cancer, and prostate cancer. The follow-up time of 55.9 (36.6, 102.5) months in the DNMs group after LT was longer than the 33.4 (18.5, 58.9) months in the non-DNMs group ( P<0.001). The 1, 5, and 10 year survival rates of patients with DNMs after LT were 96.3%, 83.5%, and 49.8%, respectively. The 1, 5, and 10 year survival rates of patients with non-DNMs after LT were 94.5%, 77.7%, and 75.4%, respectively. There was no significant difference in the cumulative survival rates between the two groups (log rank=0.402, P=0.526). Conclusion:The incidence of DNMs in LT recipients was 4.3%. The majority of them were digestive system tumors. Early diagnosis and treatment of DNMs significantly improved the prognosis and quality of life of these patients.

【Objective】 To study the screening and confirmatory test of human T-lymphotropic virus(HTLV) in blood donors. 【Methods】 Anti HTLV-1 / 2 screening was conducted on voluntary blood donors from 9 cities in Fujian province betweenJan. 12016toDec. 312018.Plasma samples ofanti-HTLV-1/2 reactive donors werecollected and sent to Xiamen Blood Center for confirmatory test. The influence of different screening reagents and confirmatory test methods on the test results were analyzed. 【Results】 A total of 741 anti-HTLV-1/2 reactivesamples were collected, among which 252 were positiveby Western Blot, 15undetermined, and 474negative, withthe overall positive rate at 34.0% (252/741). The yielding rate of domestic reagent A was significantly differentbyregions, withthe highest in Ningde(73.9%, 88/119) and the lowest in Zhangzhou(4.0%, 4/99). The confirmedpositive rates of anti-HTLV-1/2 screening reagent A(domestic) and B(imported)were 33.3%(13/39) and 57.1%(56/98), respectively, and the difference was statistically significant(P<0.01). When S / CO >5, the reagent Ayielding rate was 76.5%(13/17), significantly lower than that of imported reagent Bas 100%(56/56) (P<0.01). A total of 652 anti-HTLV-1 / 2 reactive samples were confirmed by Western Blot and nucleic acid test in parallel, among which 638 results were concordant, 14 were not, with the overall concordance rate at 97.85%. 【Conclusion】 Comparable differences in the yielding rate of twoanti-HTLV screening reagents were observed.There was over allhighconcordance, but also complementarity, between Western Blot and nucleic acid test.