Objective To establish a method for soluble fms-like tyrosine kinase-1(sFlt-1)in human serum based on magnetic nanoparticle chemiluminescence immunoassay and preliminarily evaluate its performance.Methods According to the principle of sandwich plate theory,sFlt-1 in serum reacted with biotinylated anti-sFlt-1 monoclonal antibody and acridinium ester labeled anti-sFlt-1 monoclonal antibody to form an immune complex of biotinylated antibody-antigen-acridinium ester labeled antibody.Through the reaction between biotin and streptavidin,the immune complex was captured by the magnetic nanoparticles coated with streptavidin.The acridinium ester labeled on the immune complex caused chemiluminescence in alkaline H2O2.The chemiluminescence intensity of the immune complex was used to calculate the concentration of sFlt-1 in serum,optimize the concentrations of antibodies,and establish a method for sFlt-l based on magnetic nanoparticle CLIA.The sensitivity,precision,linear range,interference,accuracy and method comparison of the established method were evaluated.A total of 100 clinical serum specimens were collected from pregnant women in the Ningbo Yinzhou Hospital of Traditional Chinese Medicine from February to August 2022.The established CLIA was used for detection,and the results were compared with the results of Roche electrochemiluminescence assay.Results The optimal concentrations of biotinylated antibody and acridinium ester labeled antibody in the reagent were ultimately determined to be 2 μ g/ml and 100 ng/ml,respectively.The limit of blank of this method was 1.67 pg/ml,the coefficient of variation(CV)of precision within and between batches was lower than 5％,and the linear range was 8.9～81 206pg/ml.When the concentrations of serum hemoglobin,bilirubin,fat emulsion and biotin were lower than 500 mg/dl,20 mg/dl,1 450 FTU and 50 μ g/L,respectively,there would be no significant interference with the detected results.The recovery rate of the method was 98％.The correlation coefficient(r2)between the established method and Roche electrochemiluminescence assay was 0.995 3.The difference between the two group was significant(P=0.000 3).Conclusion The established method have good preliminary performance and can be further applied to the auxiliary screening of preeclampsia.

Intraoperative ultrasound assisted localization is routinely used in the surgical treatment of completely endogenous renal cell tumor, however the localization and guidance ability of conventional ultrasound will decline to a certain extent for isoechoic renal tumor. A 62 years old female patient with right renal tumor was reported. The tumor diameter was about 2.3 cm× 1.7 cm, equivalent to the isoechoic of kidney. Laparoscopic partial nephrectomy was performed under the real-time guidance of contrast-enhanced ultrasound. The tumor was found to be lack of blood supply during the operation, and the tumor contour was clearly developed by contrast agent.The operation was successfully completed, and the pathological diagnosis was polycystic renal tumor with low malignant potential.The incisional margin was negative.The patient recovered well after operation without complications.No recurrence or metastasis was found after 6 months of follow-up.The renal function was good.

Objective:To study the effect of blood volume feedback control system on improving intradialytic-hypotension (IDH) in maintenance hemodialysis (MHD) patients.Methods:It was a prospective cohort study. Thirty MHD patients with recurrent IDH in the Dialysis Center of the First Affiliated Hospital of Zhejiang University School of Medicine from March 2021 to March 2022 were selected. A self-control study was conducted in MHD patients. The patients were treated with routine hemodialysis in both baseline phase (A1) and reversal phase (A2), while with hemodialysis under the blood volume feedback control system in intervention phase (B). Each phase lasted for 4 weeks (12 hemodialysis sessions). The average occurrences of IDH and IDH-related adverse events (IDH-RAE, stopping dehydration for more than 10 minutes or getting off the hemodialysis machine 10 minutes earlier due to IDH) of each patient between phase A1, B, and A2 were calculated and compared. In a total of 1 080 dialysis records, a logistic regression analysis model was established with age, sex and intervention as independent variables and with the occurrence of IDH-RAE as the outcome.Results:A total of 30 eligible patients were included in the study, including 14 males (46.7%) and 16 females (53.3%), aged 63.0 (56.5, 72.5) years old, with a median dialysis age of 84.0 (37.2, 120.0) months. The average times of IDH in 30 MHD patients decreased from 1.17 (0.83, 1.67) in stage A1 (before intervention) to 0.33 (0.25, 0.58) in stage B (after intervention) ( P<0.05). The frequency of IDH-RAE decreased significantly from 0.29 (0.19, 0.47) in stage A1 to 0.17 (0,0.25) in stage B ( P<0.05). Logistic regression analysis results indicated that the use of blood volume feedback control system reduced the risk of IDH-RAE by 53% ( OR=0.47, 95% CI 0.34-0.64, P<0.001). Conclusions:The application of blood volume feedback control system can effectively reduce the occurrences of IDH and the risk of IDH-RAE in MHD patients.

BACKGROUND: The expression of programmed cell death ligand 1 (PD-L1) as a biomarker for immunotherapy in non-small cell lung cancer (NSCLC) is routinely detected in clinical pathology department. However, the spatial heterogeneity of PD-L1 expression in intrapulmonary tumors and extrapulmonary metastases is still a challenge for the clinical testing. This study aims to explore the differences of PD-L1 expression in test samples obtaining from different sites of NSCLC. This study may contribute to the detection strategy of PD-L1 in patients with advanced lung cancer. METHODS: One hundred and thirty-one cases of consecutively detected PD-L1 (22c3 assay, Dako) staining in metastatic NSCLC and 972 cases of non-paired intrapulmonary NSCLC were collected. The discrepancies of tumor proportion score (TPS) of PD-L1 expression in intrapulmonary samples and extrapulmonary metastatic samples of different sites were compared. RESULTS: The positive expression rate of PD-L1 in extrapulmonary metastatic NSCLC (TPS ≥ 1%) was 61.83%, and the TPS was significantly higher than that in intrapulmonary tumors (P=0.03). The PD-L1 scores of the specimens obtained from different sites were significantly different (P=0.007). The positive rates of PD-L1 in liver and adrenal metastases were 85.71% and 77.78% respectively, and their TPS were significantly higher than that of the intrapulmonary samples (P<0.05). The positive rates of PD-L1 in lymph node, bone, brain, soft tissue, and pleural metastases was 40.00%-66.67%, with no significant differences compared to intrapulmonary tumors. The analysis of histological subtype and sample type showed that the PD-L1 score of extrapulmonary samples of adenocarcinoma subtype or surgical specimen was significantly higher than that of intrapulmonary tumors. The analysis of clinicopathological parameters showed that the PD-L1 positive expression or high expression were significantly correlated with male patients, smoking history, and epidermal growth factor receptor (EGFR) wild type. CONCLUSIONS The expression of PD-L1 in metastatic NSCLC is generally higher than that in intrapulmonary tumor, and the positive rate of PD-L1 expression was discrepant in different sites of specimen. The differences of PD-L1 score between extrapulmonary metastatic samples and intrapulmonary samples may be associated with different metastatic sites, histological subtype, and specimen type.
B7-H1 Antigen/metabolism* ; Biomarkers, Tumor/metabolism* ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Humans ; Immunohistochemistry ; Lung Neoplasms/drug therapy* ; Male

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.

IgG4-related diseases have a low incidence and are easily misdiagnosed as tumors in clinical treatments. A 26-year-old male patient was admitted to the hospital because of a left adrenal tumor found in health examination for more than 5 months. The tumor in the left adrenal region could be seen from abdominal CT, and the retroperitoneal laparoscopic resection of the left adrenal tumor was performed. Postoperative pathology was consistent with IgG4-related diseases, and serum IgG4 was abnormally high. After 2 months’ follow-up, serum IgG4 returned to normal, and no special discomfort.

The incidence of solitary fibroma of seminal vesicle is low, and the source of seminal vesicle is rare. A 38-year-old patient was admitted to hospital because of intermittent gross hematuria for more than one month. Seminal vesicle space occupying lesions can be seen in pelvic MRI. Laparoscopic resection of right seminal vesicle tumor was performed, and the pathological diagnosis tended to solitary fibroma. During the 5-month follow-up, the symptoms of hematuria disappeared and no recurrence.

BK polyomavirus-associated nephropathy (BKPyVAN) is a common cause of allograft failure. However, differentiation between BKPyVAN and type I T cell-mediated rejection (TCMR) is challenging when simian virus 40 (SV40) staining is negative, because of the similarities in histopathology. This study investigated whether donor-derived cell-free DNA (ddcfDNA) can be used to differentiate BKPyVAN. Target region capture sequencing was applied to detect the ddcfDNAs of 12 recipients with stable graft function, 22 with type I TCMR, 21 with proven BKPyVAN, and 5 with possible PyVAN. We found that urinary ddcfDNA levels were upregulated in recipients with graft injury, whereas plasma ddcfDNA levels were comparable for all groups. The median urinary concentrations and fractions of ddcfDNA in proven BKPyVAN recipients were significantly higher than those in type I TCMR recipients (10.4 vs. 6.1 ng/mL,

Objective To compare the clinical outcomes of low-dose rabbit antithymocyte globulin (rATG ) vs basiliximab as induction therapy in recipients of ABO-incompatible kidney transplantation (ABOi-KT) .Methods Retrospective analysis was conducted for e the clinical data of 40 ABOi-KT recipients between March 2017 and March 2019 .17 recipients of them received induction therapy with basiliximab (basiliximab group) while another 23 recipients received low-dose rATG (rATG group ,rATG 25 mg/d × 3 d) .During a median follow-up period of 282 days , the data of serum creatinine and eGFR at 1 week and 1 month ,graft survival rate and complication rate of two groups were compared .Results No significant difference existed in age ,gender ,dialytic modality/ duration , blood groups of recipients , HLA mis-match , blood group antibody titers , dose of rituximab ,blood groups of donors or donor age ( P > 0 .05 ) . The times of double filtration plasmapheresis in Basiliximab group were more (P< 0 .05) .No significant difference existed in serum creatinine and eGFR at 1 week or 1 month ( P > 0 .05 ) . No significant difference existed in graft survival rate . No significant difference existed in rate of acute rejection ,parvovirus B19 infection , urinary tract infection or hematoma .Conclusions Low-dose of rATG is as effective as basiliximab for ABOi-KT recipients .And rATG does not increase the rate of infection .

Objective To analyze the feasibility and safety in application of resectoscope combined with laparoscopy in the operation treatment of paraganglioma of urinary bladder.Methods 7 cases patients with paraganglioma of urinary bladder treated in our hospital from November 2014 to August 2018 were analyzed retrospectively.There were 5 males and 2 females,average age of 31.1 years (22-37 years),average body mass index was 22.3 kg/m2 (18.3-22.5 kg/m2).All the 7 cases patients complained of dizziness and palpitation after urination,average basal systolic blood pressure was 111.8 mmHg (97-124 mmHg),the average fluctuation of systolic blood pressure before and after urination was 64.9 mmHg(28-91 mmHg),the CT and cystoscopy prompt bladder tumor,the average diameter was 2.7 cm(2.1-3.5 cm).The average of plasma norepinephrine was 706.3 pg/ml(330-997 pg/ml);the average of plasma dopamine was 101.1 pg/ml(44-145 pg/ml);the average of 24h urinary vanilmandelic acid was 13.4 mg/24h (10.3-16.1 mg/24h).All the patients has controlled the blood pressure and dilate the blood vessels with phenoxybenzamine hydrochloride,accepted the operation of resectoscope combined with laparoscopy partial cystectomy and bladder sutura per abdomen after ample dilatancy.The patients had lithotomy position with trendelenburg,preparation of gas peritoneal cavity by transabdominal,inside the resectoscope by transurethral at the same time,mutilated bladder mucosa beside 1cm at the edge of the tumor,and cut full thickness bladder wall,take the extraperitional fat as the standard procedure;we could see the cutting edge clear at this time by laparoscopy,cut off the pelvic peritoneum,extraperitional fat and the tumor.The sample placed in bladder,close the bladder with absorbable or barbed wires,take out the sample by resectoscope.Results All the 7 cases patients operation was successfully completed,no cases has been transfered to open.The average time of operation is 85.3 min(65-100 min),the average amount of bleeding is 27.9 ml(10-50 ml).The average fluctuation of systolic blood pressure is 8.7 mmHg(6-15 mmHg).Bladder washout was stopped 24h after operation,catheter was removed 1 weeks after operation.There is no obvious complications occurred.The average hospital stay is 3.7 days (3-5 days).The average pain score of 4 cases 4h after operation is 3.8 (2-5),reevaluation 24h after operation is 2.3 (1-4).The average follow-up time is 7.9 months(2-15 months).All the 7 cases patients clinical symptoms disappeared,there is no fluctuation of systolic blood pressure before and after urination,there is no recurrence of the tumor.Conclusions To the paraganglioma of urinary bladder in fundus of bladder or anterior wall of bladder,we can accurate resection tumor by resectoscope combined with laparoscopy,reduce blood pressure fluctuations,reduce the surgical trauma and the distress of patients.It is a safety and effective minimally invasive surgery.