[Analysis of BK virus infection in kidney transplant recipients].

Objective: To compare the infection of BK virus in the recipients of living donor(LD) kidney transplant and deceased donor(DD) kidney transplant. Methods: A total of 911 recipients who underwent kidney transplantation in the Organ Transplantation Research Institute of the 8th Medical Center of the People's Liberation Army General Hospital from January 2015 to August 2019 were enrolled in this study. The DNA copies of BK virus in urine and peripheral blood of kidney transplant recipients were detected by real-time quantitative PCR. The patients were divided into LD group (n=255) and DD group (n=656). BK virus infection in recipients with DD kidney transplant were compared with that in recipients of LD kidney transplant. Results: The BK virus positive rate in the urine of all subjects was 13.06%(119/911), and that in blood was 2.96% (27/911). The positive rate of BK virus in urine after kidney transplantation was significantly higher than that in blood(P<0.000 1). The positive rate in urine was 9.02% (23/255) in LD group, which was significantly lower than that of 14.63% (96/656) in DD group in the same period (χ(2)=5.097, P=0.012); The positive rate of BK virus infection in relatives group was 0.78% (2/255), which was significantly lower than that of 3.81% (25/656) in DD group (χ(2)=5.849, P=0.007). Conclusions: There was a significant difference in the infection rate of BK virus between the LD and DD group. The incidence of BK virus infection in kidney transplant recipients from DD was higher than that of from LD kidney transplant recipients.

[Cytomegalovirus active infection after kidney transplantation affecting the expression of activation receptor CD226 on NKT cell].

Objective: To investigate the effect of active cytomegalovirus infection post kidney transplant on the expressing of receptor CD226 on NKT cell. Methods: Case controlled study. From December 2013 to December 2014, 43 cases of kidney transplant recipient with CMV infection were collected in the Organ Transplantation Research Institute of the former 309th Hospital of PLA. The healthy control group included 15 cases. 15 cases of recipients who were stable after operation and followed up in our hospital at the same time were also collected as control. Peripheral blood specimen with EDTA as anticoagulant were used and analyzed by flow cytometry. Results: The population of NKT in CMV infection recipients were 5.19(1.18, 25.92)%, while in the remission stage the population were 4.89(0.68, 25.33)%, Compared with normal healthy controls and the stable recipients, the percentage of CD3(+)CD56(+) NKT cells in periphery blood mononuclear cells did not vary among these groups(P>0.05). The CD226(+) NKT population during the active CMV infection was (70±13)%, which was significantly lower than the health control [(87±10)%] and stable recipients [(80±9)%](P<0.001). Whereas in the CMV infection remission stage, the CD226(+)NKT population was (81±16)%, which was significantly higher than that of CMV active infection group (P<0.05), and showed no difference with the health control group and stable recipients (P>0.05). The CD226 MFI expressed on NKT in CMV infection group was 101±49, which showed no difference with health controls and stable recipients (P>0.05). However, significantly up regulation of CD226 MFI on NKT was observed in the samples obtained from the same recipients in CMV active infection (91±40) and in CMV regression stage(173±73)(P<0.001). Conclusions: The CD226(+) NKT cells population was down during the active CMV infection post kidney transplantation, while the expression of CD226 and the population of CD226(+)NKT could regression when the CMV infection regressed, which indicates the involvement of CD226 in the process of NKT cells anti-CMV infection.

[A preliminary research on the exosome influenced by virus infection in the renal transplantation recipients].

Objective: Virus infection is a common complication of transplantation.With the research and application of exosome is becoming more popular, this study focused on whether the virus particles and nucleic acids exist in the exosomes extracted from the plasma of recipients with virus infection after renal transplantation. Methods: A total of 10 independent transplantation recipients at Institute of Organ Transplantation, 309th Hospital of Chinese People's Liberation Army from January 2015 to July 2017 were studied in this study.5 cases of positive or suspected positive in granulocytes HCMV pp65 antigen detection and positive in plasma HCMV DNA test, and the other 5 cases of positive results in plasma BK DNA test were adopted.Exosomes were extracted from the collected plasma samples with SBI kit.Electron microscopy and nanoparticles tracing analyzer (NTA) were used for exosome analysis.Quantitative real-time PCR method was used to inspect and compare virus DNA copies number in plasma, exosome and effluent. Results: Typical exosome-like vesicle structure was observed.NTA put forward the sample concentration data from 1.2 to 4.5×10(12) particles/ml, and the particle diameters were 30-200 nm.In the qRT-PCR assays, the viral DNA quantitative results of exosome samples are lower but on the same magnitude compared with that of the plasma, and sharply decreased in effluent. Conclusions: Virus DNAs in exosome samples of recipients with viral infection after transplantation were detected in great quantities.This not only hints the spread of the virus may take advantage of the biological formation process of exosomes, but also warns that the limitation of the existing way to extract exosmes from virus infected population may be a bottleneck in research.

[Research of mHLA-G and the receptor expression with kidney rejection and CMV active infection following kidney transplantation].

Objective: To study the expression of membrane HLA-G (mHLA-G) and the receptor immunoglobulin-like transcript 2(ILT2) on lymphocyte and find their association with rejection and cytomegalovirus (CMV) infection after renal transplantation. Methods: A total of 88 cases of renal transplant recipients for the first time from February 2014 to February 2016 were studied in this work. Recipients can be divided into rejection group (n=12) and stable renal function group (n=41) according to whether rejection occurred. Recipients only infected CMV not developed rejection were included in the CMV positive group (n=24). CMV negative group (n=11) including CMV negative recipients once infected CMV.The expression of mHLA-G and ILT2 on lymphocytes were detected by flow cytometry, and the differences among different groups were analyzed. Results: The data showed that after renal transplantation, T and B lymphocytes mHLA-G expression rate was the lowest in the rejection group (0.42%±0.35%, 0.88%±0.47%), having significant difference with renal function stable group and CMV positive group (all P<0.01). In CMV positive group the expression of mHLA-G on T and B lymphocytes was the highest (1.31%±0.69%, 2.01%±0.91%), having significant difference with rejection group (P<0.001). The expression of mHLA-G on B cell was statistically significantly different between CMV positive group and CMV negative group (P<0.05). There was no significant difference in ILT2 expression on B cell among the four groups (P>0.05). The expression rate of ILT2 on T cells was higher in the CMV positive group (36.91%±14.91%), having significant difference with the other three groups (P<0.01). Conclusions: Low expression of mHLA-G on T and B lymphocytes may predict rejection after renal transplantation. High expression of mHLA-G and ILT2 on lymphocytes is prone to CMV infection after renal transplantation .

[Expression and significance of neutrophil surface adhesion molecules in renal transplant recipients with cytomegalovirus infection].

OBJECTIVE: To study the expression and its diagnostic significance of neutrophil surface adhesion molecules including CD11b, CD15 and CD62L after renal transplantation in recipients with cytomegalovirus (CMV) infection. METHODS: Blood samples were collected from 142 kidney transplant recipients, including 95 males and 47 females, who received allogeneic renal transplantation between September 2009 and January 2015 in 309th Hospital of the PLA. Healthy volunteers (22 males and 9 females) were recruited from physical examination center in 309th Hospital of the PLA from September 2009 to January 2015 as healthy control group. Renal transplant recipients were divided into high active CMV infection group, active CMV infection group and CMV negative control group according to CMV-pp65 antigen detection. Neutrophil surface adhesion molecules CD11b, CD15 and CD62L were detected by flow cytometry and their mean fluorescence intensity compared among the groups. Receiver operating characteristic (ROC) curves of CD11b, CD15 and CD62L in detecting active infection in renal transplant recipients were made. RESULTS: The mean fluorescence intensity of CD15 in high active CMV infection group(n=17) and active CMV infection group(n=65)were 776.31±89.53 and 554.39±67.89, respectively, with significant differences compared with CMV negative control group (n=60, 334.92±44.69) and healthy control group (n=31, 310.56±39.67) (all P<0.05); the expression proportions of CD11b and CD62L in high active CMV infection group and were 42.31%±6.11% and 40.35%±6.47%, respectively, with significant differences compared with active CMV infection group(62.45%±5.67% and 65.65%±5.33%), CMV negative control group(70.74%±6.55% and 70.37%±6.71%) and healthy control group(72.52%±6.48% and 72.43%±6.51%) (all P<0.05). The optimal cut-off values of CD11b and CD62L in diagnosing active CMV infection group were 56.61% and 44.35%, respectively, with the sensitivity being both 100.00%, the specificity being 76.67% and 58.06% respectively, and the area under the curve (AUC) being 0.851 and 0.628 respectively; the optimal cut-off values of CD11b and CD62L in diagnosing high active CMV infection group were 66.57% and 69.56% respectively, with the sensitivity being 81.54% and 87.69% respectively, the specificity being 100.00% and 98.33% respectively, and the AUC being 1.000 and 0.991 respectively; the optimal cut-off values of mean fluorescence intensity of CD15 in diagnosing high active CMV infection group and active CMV infection group were 542.71 and 408.03 respectively, the sensitivity in the two groups being 100.00% and 98.46% respectively, the specificity being both 100.00%, and the AUC being 1.000 and 0.999 respectively. CONCLUSIONS: Neutrophils CD15 expression may be up-regulated in renal transplantation recipients with CMV infection, while neutrophils CD11b and CD62L expressions are down-regulated. Such changes in CD15, CD11b and CD62L expression can be used as a basis for laboratory diagnosis of active CMV infection.

A 10-year retrospective study of primary hyperparathyroidism in children.

OBJECTIVE: Primary hyperparathyroidism (PHPT) in children is thought to be extremely rare. The exact incidence is unknown and little is known of the characteristics of the disease. We reviewed our experience with PHPT to better characterize these patients. METHODS: We carried out a retrospective study in 12 patients (age 9-16 years) who had surgical treatment between 2001 and 2011 in a single institution. RESULTS: There were 5 male subjects and 7 female subjects. The diagnosis of PHPT in pediatric patients is frequently delayed with an average time of 41 months. The main signs of PHPT were urinary and bone disorders, as well as non-specific signs. All patients had preoperative localization studies prior to operation. Combined using of neck sonography, MIBI-scintigraphy and CT and/or MRI, our patients had a positive predictive value of 100%. After operation, all patients were confirmed to have parathyroid adenoma. 1 patient had 2 adenomas, 1 of which was an ectopic adenoma located in the mediastinum. 9 patients experienced symptomatic hypocalcemia postoperatively. No patients had any serious long-term effects from parathyroidectomy at our institution. All patients were cured after removal of the parathyroid adenoma. No permanent complication was observed for all patients. CONCLUSION: PHPT in children is rare, the commonest signs are urinary and bone tissue impairment, as well as non-specific signs. Evaluation of serum calcium and PTH levels is diagnostic in suspected patients. Preoperative localization is essential. Surgery is without significant complications. Surgical treatment of PHPT is curative and definitive.