Predictive values of donor TIMP-2 and IGFBP7 in delayed graft function after kidney transplantation / 中华器官移植杂志

Objective:To explore the predictive values of tissue inhibitor of metalloproteinase-2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) in donor sera and lavage fluid on delayed graft function (DGF) in donation after circulatory death (DCD) kidney transplant recipients.Methods:A total of 33 eligible kidney donors and 33 corresponding recipients were recruited. Preoperative serum and renal perfusion fluid samples of donors were collected to determine the levels of TIMP-2 and IGFBP7. Patients were grouped according to whether DGF occurred after kidney transplantation and measured indicators analyzed. Independent sample t test was utilized for comparing the groups with normal distribution measurement data. And χ2 test was employed for comparing the groups with normal distribution counting data and Mann-Whitney test for comparing the groups with non-normal distribution measurement data. Receiver operating characteristic (ROC) curve and area under curve (AUC) were used for evaluating the diagnostic efficacy of indicators. Results:In donor-DGF group, lavage fluid TIMP-2, product of lavage fluid TIMP-2 and IGFBP7 (TIMP-2×IGFBP7), serum IGFBP7 and product of serum TIMP-2 and IGFBP7 (TIMP-2×IGFBP7) were higher than those in donor-non-DGF group ( P<0.05). The AUC of TIMP-2, TIMP-2×IGFBP7, serum IGFBP7 and serum TIMP-2×IGFBP7 in the diagnosis of DGF were 0.753 (95%CI 0.546~0.959), 0.747 (95%CI 0.510~0.984), 0.824 (95%CI 0.615~1.000) and 0.852 (95%CI 0.660~1.000) respectively. Conclusions:Donor serum IGFBP7, donor serum TIMP-2×IGFBP7, lavage fluid TIMP-2 and lavage fluid TIMP-2×IGFBP7 may be used for predicting the occurrence of early DGF after kidney transplantation. Among them, serum TIMP-2×IGFBP7 has the highest diagnostic efficiency and may be an excellent predictor of DGF occurrence.

The predictive value of TIMP-2 and IGFBP7 in delayed renal function recovery after DCD renal transplantation / 中华泌尿外科杂志

Objective:This study is to investigate the predictive value of serum levels of TIMP-2 and insulin-like growth factor-binding protein 7(IGFBP7) in patients with DCD(donation after cardiac death) kidney transplantation.Methods:A prospective research design was used to select DCD kidney transplant patients admitted to the Li Huili Hospital of Ningbo University from January 2018 to October 2020.Inclusion criteria: ①Complete data; ②There were no serious complications affecting the function of the transplanted kidney in the early postoperative period.Exclusion criteria: ①Incomplete data; ②Patients were unable or unwilling to cooperate with the study; ③Severe complications affecting the function of the transplanted kidney occurred early after the operation.The ELASE method was used to quantitatively detect the serum TIMP-2 and IGFBP7 levels at 6, 12, 24, 48, 72 hours and 7 days after renal transplantation, and monitor the serum creatinine values during the same period and 21 days after the operation. According to the occurrence of DGF, the measured values of TIMP-2 and IGFBP7 at different time points and their product's ability to predict the occurrence of DGF after kidney transplantation were analyzed. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the diagnostic efficacy of TIMP-2 and IGFBP7 for DGF.Results:A total of 33 patients were enrolled, 7 patients (21.2%) in the DGF group and 26 patients (78.8%) in the non-DGF group. Between the two groups, the donor glomerular filtration rate were [98.5(15.8-132.5)ml/(min·1.73m 2) and 79.1(60.6-102.5)ml/(min·1.73m 2)], recipient gender (male/female: 3/4 cases and 10/16 cases), recipient age [48(34-56) Years old and 45(23-61) years old], the recipient's preoperative creatinine [1114.0(731.4-1293.0)μmol/L and 858.4(657.6-1051.9)μmol/L], the recipient's preoperative urea nitrogen [15.0(13.2-19.6)mmol/L and 17.3(13.6-20.9)mmol/L], receptor preoperative albumin [43.5(38.5-45.3)mmol/L and 41.2(37.5-46.1) mmol/L], recipient dialysis method [hemodialysis/peritoneal dialysis: 3/4 cases and 9/17 cases], warm ischemia time [6(5-7) and 5(4-6) min, there was no statistically significant difference] ( P>0.05). The values of serum IGFBP7 and TIMP-2×IGFBP7 in the DGF group were higher than those in the non-DGF group at all time points ( F=15.753, P=0.040; F=13.000, P=0.024), while serum TIMP-2 was not significant between the two groups difference ( F=1.157, P=0.075). For the diagnostic value of DGF, the AUC of serum IGFBP7 at 48 h after surgery was 0.863 (95% CI 0.696-1.000, P=0.004). When 5.97 ng/ml was used as the cut-off value, the sensitivity was 85.7% and the specificity was 80.8 %. The AUC of TIMP-2×IGFBP7 at 48 hours after surgery was 0.819 (95% CI 0.641-0.996, P=0.011). When 62.06(ng/ml) 2 was used as the cutoff value, the sensitivity was 71.4% and the specificity was 80.8%.There was no statistical difference in the area under the curve between the two ( P>0.05). There were differences in the dynamic trend of serum IGFBP7 and creatinine in the DGF group. Serum IGFBP7 at 7 days after surgery was positively correlated with creatinine at 21 days after surgery. Conclusion:Serum IGFBP7 and TIMP-2×IGFBP7 could predict the occurrence of DGF after DCD donor kidney surgery. The predictive value changes with time. Among them, 48h and 7d after surgery are the most valuable. However, serum TIMP-2 has not been found to have predictive value in this study.

Survey of RBRVS-based performance appraisal methods at 12 public hospitals / 中华医院管理杂志

Objective To learn the initial application of RBRVS related performance appraisal at public hospitals in China. Methods 12 tertiary public hospitals which took the lead to introduce RBRVS models, via third-party cooperation, in medical stuff performance assessment over one year were surveyed. The study summarized their design emphases of RBRVS performance appraisal, covering such aspects as appraisal level, accounting method, conceptual design, points setup and items charged as cost. Descriptive statistics was used to analyze the data so acquired. Results As shown in the performance appraisal level, most hospitals were found with defects in refining levels, as eight hospitals only appraise down to the department level, three to diagnostic group level, and only one hospital to individuals. Of these hospitals, over eight hospitals used RBRVS to elevate the efficiency and inventiveness of employees, while half of these hospitals reported the method can improve service, reduce cost and enhance quality of care. Conclusions Public hospitals enjoy advantages via third-party cooperation RVRBS method in RVRBS-related reforms. Introduction of this method calls for localization design based specifics and management goals of public hospitals.

RBRVS-based performance appraisal: application at a hospital / 中华医院管理杂志

With the resource based relative value scale (RBRVS) application at a hospital cited as an example, the paper covered its use in performance appraisal for optimal performance-based distribution as follows. In view of hospital specifics, such appraisal indexes as the quantity, quality, technical difficulty and cost control are quantified as key indicators, to form its own RBRVS scores and weights. In combination of the case mixed index as well, such factors as specific appraisal, quality of care and research/teaching work are taken into account for the purpose of rational and optimal pay for performance.

Proteomic analysis of nasal lavage fluid in patients with allergic rhinitis / 国际检验医学杂志

Objective To screen the biomarkers in nasal lavage fluid of patients with allergic rhinitis (AR) by proteomics tech‐nology .Methods The nasal lavage fluid samples of 10 cases of AR patients and 20 cases of normal controls were collected both be‐fore and after allergen stimulation tests .Two‐dimensioned gel eletrophoresis was used to detect the proteins in nasal lavage fluid samples .The proteins that showed different expression levels were identified by MALDI‐TOF/TOF mass spectrometry .Results Six kinds of proteins differentially expressed were screened out .The expressions of lactoferrin ,α1‐antitrypsin ,and APOA1 increased in the AR patients after being stimulated by allergens .On the contrary ,the expressions of PLUNC protein ,haptoglobin and IgJ de‐creased .Conclusion The six kinds of proteins might be the biomarkers of AR .

Rapid expression and preparation of the recombinant fusion protein sTNFRII-gAD by adenovirus vector system / 生物工程学报

We expressed and prepared the recombinant fusion protein sTNFRII-gAD consisted of soluble TNF receptor II and the globular domain of adiponectin by Adenovirus Vector System in mammalian BHK21c022 cells. First we used the adenovirus vector containing EGFP gene (rAd5-EGFP) to infect BHK21c022 cells at different MOI (from 0 to 1 000), and then evaluated their transduction efficiency and cytotoxicity. Similarly, we constructed the replication-deficient adenovirus type 5-sTNFRII-gAD (rAd5-sTNFRII-gAD). We collected the supernatants for Western blotting to determine the optimal MOI by comparing the expression levels of sTNFRII-gAD fusion protein, 48 h after the BHK21c022 cells were infected by rAd5-sTNFRII-gAD at different MOIs (from 0 to 1 000). Then, we chose rAd5-sTNFRII-gAD at MOI 100 to infect five bottles of BHK21c022 cells in 100 mL of serum-free chemically defined media 100 mL, harvested the supernatant every 48 h for 6 times, and condense and purify sTNFRII-gAD fusion protein by ammonium sulfate salt-out and size-exclusion chromatography, respectively. Finally, we analyzed anti-TNFalpha activity of sTNFRII-gAD fusion protein on L929 cells in vitro. The results showed that the number of BHK21c022 cells expressing EGFP protein was increased significantly with the increase of MOI. However, some cells died at MOI of 1 000 while there was no significant cytotoxicity at MOI from 0 to 100. Western blotting analysis showed that the more adenoviruses, the higher expression of sTNFRII-gAD fusion protein in the supernatant with the highest expression at MOI 1 000. We successfully obtained about 11 mg bioactive and purified sTNFRII-gAD fusion protein at last. The in vitro assay demonstrated that the sTNFRII-gAD fusion protein was potent to antagonize TNFalpha's cytotoxicity to L929 cells. Put together, we established a recombinant adenovirus vector/BHK21 cell expression system, characteristic of the efficient serum-free culture and easy scaling-up.

A Synchronistic Deficiency in Expression of HLA Class I Molecules and TAP/LMP Genes in Human Ovarian Cancer Cells / 中国肿瘤生物治疗杂志

Objective: Full expression of class I HLA molecules on tumor cells is pivotal in priming the tumor antigen-specific CTLs for effective iramunotherapies. Tumor cells display abnonnal expression of HLA class I molecules in human ovarian carcinoma. Methods: In this study, the expression of class I molecules in human ovarian cancer cells was determined by using Western bloting, immunohistochemistry testing and flow cytometry. The mRNA of TAP (transporter associated with antigen processing) and LMP(low molecular weight polypeptide) were exannined at the same time by RT-PCR. Results: It has been shown that class I molecules were unable to be, or only weakly, expressed in five of eight ovarian cancer cell lines. On these cell lines abnormal in class I expression, no or only little mRNA were detected for TAP1, TAP2, LMP2 and(or) LMP7 genes. Class I expression, however, could be partially recovered by incubating the tumor cells at 25℃ when compared with those at 37℃ . Conclusion: The results suggested that the dysfunction of TAP and IMP genes, following by a defective expression of class I molecules, might be one of the mechanisms enable ovarian cancer cells to escape from immune surveillance.

Construction of cDNA Library of Ovarian Carcinoma / 中国肿瘤生物治疗杂志

We isolated polyA~+ mRNA direcdy from tumor tissue of ovarian carcinoma using Oligotex~(TM) Direct mRNA Kit (QIAGEN) to synthesize the first and second strand cDNA. The ds-cDNA termini were blunted with pfu DNA poly-rnerase. The blunted cDNAs were added EcoR I adaptor, and then were digested by Xho I . Small cDNA molecules(

Cloning of HLA-A~*0201 and its transient expression on COS-7 cells / 中国免疫学杂志

Objective:To clone HLA A *0201 cDNA and express it transiently on COS 7 cells.Methods:HLA A cDNA isolated from the HLA A *0201 positive lymphocytes by using RT PCR was cloned into pBluescript II SK vector directionally.After the sequence was confirmed,the cDNA was inserted into mammalian expression vector pcDNA3.The recombinant vector was transfected into COS 7 cells by cation liposome.The transient expression on the cells was measured by flow cyteometer.Results:The cDNA was identical with HLA A *0201 cDNA published on Genebank.Determined by flow cytemeter,the expressing rate was recorded for 57%.Conclusion:We have cloned HLA A *0201 successfully and expressed it transiently on COS 7 cell,which would be potentially useful in research on killing tumor restricted by HLA A2.