[Colorectal cancer with ß-catenin protein expression deficiency: a clinicopathological analysis].

Objective: To investigate the clinicopathological features and molecular characteristics of ß-catenin-deficient colorectal cancer. Methods: The clinical, pathological and molecular features of 11 colorectal cancers with ß-catenin protein loss diagnosed at the 960th Hospital of People's Liberation Army of China, from January 2012 to November 2022 were analyzed. Results: Among the 11 patients, 3 were males and 8 were females. Their age ranged from 43 to 74 years, with the median age of 59 years. Six were in the left colon and 5 were in the right colon. One of the 11 cases had lymph node metastasis, 10 cases were well and moderately differentiated adenocarcinoma, and 1 was mucinous adenocarcinoma. Eight cases were of TNM stage T4, 2 of T1 stage and 1 of Tis stage. ß-catenin protein was not detected using immunohistochemistry. Sanger sequencing revealed the presence of fragment-deletion mutation in exon 3 of CTNNB1 gene, resulting in loss of ß-catenin protein expression. Conclusion: ß-catenin deficiency is present in a small number of colorectal cancers and may be associated with exon 3 mutations of CTNNB1 gene.

[Analysis of the clinical characteristics of acute myeloid leukemia related to the treatment of hematological and solid tumors].

Objective: To compare and analyze the clinical characteristics of acute myeloid leukemia (AML) related to the treatment of hematological tumors and solid tumors. Methods: The laboratory and clinical data of 41 patients with treatment-related AML (t-AML) in the Department of Hematology, Henan Cancer Hospital from January 2014 to December 2021 were retrospectively analyzed, and they were divided into hematological tumor group and solid tumor group. Survival analysis was performed using the Kaplan-Meier method and Log rank test. Results: The median interval from the first tumor diagnosis to t-AML in 41 patients was 21.0 (16.5-46.0) months; 24 (58.5%) had abnormal expression of lymphoid antigen, 28 (68.3%) had abnormal karyotype, 18 cases (43.9%) were positive for fusion gene, and 28 cases (68.3%) were positive for gene mutation; the median recurrence-free survival (RFS) was 11.0 months, and the median overall survival (OS) was 11.5 months. The proportion of acute promyelocytic leukemia ([APL], 0.0, 0/13), complete response ([CR],18.2%, 2/11), median OS (4.5 months) and median RFS (2.5 months) of t-AML patients in the hematological tumor group were significantly lower than those in the solid tumor group (35.7%, 10/28; 68.0%, 17/25; not reach; not reach), but the proportion of M4 /M5 (93.2%,12/13) was significantly higher than that in the solid tumor group (53.6%,15/18; all P values<0.05). Through subgroup analysis, the proportion of patients with positive PML-RARa and good prognosis karyotypes in the solid tumor group (35.7%, 10/28; 46.4%, 13/28) was significantly higher than that in the hematological tumor group (0.0, 0/13; 0.0, 0/13; P<0.05), while the proportion of patients with intermediate karyotypes (42.9%, 12/28) was significantly lower than that in the hematological tumor group (84.6%, 11/13; P<0.05), the difference was statistically significant. The CR rate (90.0%, 9/10), median OS (not reach) and median RFS (not reach) in the t-APL group were higher than those in the t-AML (without t-APL) group (38.5%, 10/26; 6 months; 8 months; P<0.05). After excluding the effect of t-APL patients, there was no significant difference in the CR rate, median OS and median RFS between the solid tumor group (8; 9 months; not reach) and the hematological tumor group (2; 4 months; 2 months; P>0.05). Univariate analysis showed that the primary tumor belongs to hematological tumor was a common risk factor for OS and RFS in t-AML patients (P<0.10). Conclusions: Compared with patients with t-AML secondary to solid tumors, patients with t-AML secondary to hematological tumors have poorer treatment effects and poorer prognosis. After excluding the effect of t-APL patients, there are no significant differences in the treatment efficacy and prognosis between the two types of t-AML patients.

[Association of circulating levels of soluble PD-1, PD-1 gene polymorphisms with HBV infection and HBV infection-associated hepatocellular carcinoma].

Objective: To investigate the association of circulating sPD-1 level and PD-1 gene polymorphisms with HBV infection and HBV infection-associated hepatocellular carcinoma. Methods: A case-control study was conducted. A total of 237 chronic HBV infection cases and 138 HBV infection-associated hepatocellular carcinoma in the Department of Infectious Diseases of the First Hospital of Shanxi Medical University from 2018 to 2021 were selected as the case group. About 250 individuals who visited a hospital physical examination center for routine physical examination during the same period were selected as the control group. Plasma sPD-1 levels were measured by using an ELISA kit and genotyping was performed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The association of sPD-1 levels and PD-1 polymorphisms with HBV infection as well as HBV infection-associated hepatocellular carcinoma was analyzed by using logistic regression models after adjusting for age, sex, alcohol consumption, smoking, ALT and AST levels. The sPD-1 level and PD-1 polymorphisms were independent variables, and HBV infection was the dependent variable. Results: The age of 237 chronic HBV infections, 138 HBV infection-related liver cancer case subjects and 250 control subjects in the study was (49.1±10.8), (51.9±12.7) and (50.7±11.9) years, respectively. Multivariate logistic regression model analysis showed that with a 1 pg/ml increase in sPD-1 level, the OR (95%CI) values for the risk of incident HBV infection cases and HBV hepatocellular carcinoma cases were 1.92 (1.68-2.19) and 2.02 (1.69-2.40). For rs2227981, compared with the CC genotype, the TT genotype had a lower risk of HBV infection and liver cancer associated with HBV infection, with OR (95%CI) values of 0.45 (0.22-0.91) and 0.35 (0.14-0.91). For rs2227982, compared with the CC genotype, the CT and TT genotypes also had a lower risk of HBV infection [OR (95%CI) values of 0.72 (0.53-0.97) and 0.57 (0.35-0.93)] and HBV infection-related liver cancer [OR (95%CI) values of 0.64 (0.45-0.92) and 0.52 (0.29-0.93)]. Conclusions: Plasma sPD-1 levels and PD-1 gene polymorphisms are associated with HBV infection and HBV infection-associated hepatocellular carcinoma.

Population Pharmacokinetics of Intranasal Dexmedetomidine in Infants and Young Children.

BACKGROUND: Intranasal dexmedetomidine provides noninvasive, effective procedural sedation for pediatric patients, and has been widely used in clinical practice. However, the dosage applied has varied fourfold in pediatric clinical studies. To validate an appropriate dosing regimen, this study investigated the pharmacokinetics of intranasal dexmedetomidine in Chinese children under 3 yr old. METHODS: Intranasal dexmedetomidine 2 µg · kg-1 was administered to children with simple vascular malformations undergoing interventional radiological procedures. A population pharmacokinetic analysis with data from an optimized sparse-sampling design was performed using nonlinear mixed-effects modeling. Clearance was modeled using allometric scaling and a sigmoid postmenstrual age maturation model. Monte Carlo simulations were performed to assess the different dosing regimens. RESULTS: A total of 586 samples from 137 children aged 3 to 36 months were included in the trial. The data were adequately described by a two-compartment model with first-order elimination. Body weight with allometric scaling and maturation function were significant covariates of dexmedetomidine clearance. The pharmacokinetic parameters for the median subjects (weight 10 kg and postmenstrual age 101 weeks) in the authors' study were apparent central volume of distribution 7.55 l, apparent clearance of central compartment 9.92 l · h-1, apparent peripheral volume of distribution 7.80 l, and apparent intercompartmental clearance 61.7 l · h-1. The simulation indicated that at the dose of 2 µg · kg-1, 95% of simulated individuals could achieve a target therapeutic concentration of 0.3 ng · ml-1 within 20 min, and the average peak concentration of 0.563 ng · ml-1 could be attained at 61 min. CONCLUSIONS: The pharmacokinetic characteristics of intranasal dexmedetomidine were evaluated in Chinese pediatric patients aged between 3 and 36 months. An evidence-based dosing regimen at 2 µg · kg-1 could achieve a preset therapeutic threshold of mild to moderate sedation that lasted for up to 2 h.

Dual Functions of MDP Monomer with De- and Remineralizing Ability.

Methacryloyloxydecyl dihydrogen phosphate (MDP) has been speculated to induce mineralization, but there has been no convincing evidence of its ability to induce intrafibrillar mineralization. Polymers play a critical role in biomimetic mineralization as stabilizers/inducers of amorphous precursors. Hence, MDP-induced biomimetic mineralization without polymer additives has not been fully verified or elucidated. By combining 3-dimensional stochastic optical reconstruction microscopy, surface zeta potentials, contact angle measurements, inductively coupled plasma-optical emission spectroscopy, transmission electron microscopy, atomic force microscopy, and Fourier transform infrared spectroscopy with circular dichroism, we show that amphiphilic MDP can not only demineralize dentin by releasing protons as an acidic functional monomer but also infiltrate collagen fibrils (including dentin collagen), unwind the triple helical structure by breaking hydrogen bonds, and finally immobilize within collagen. MDP-bound collagen functions as a huge collagenous phosphoprotein (HCPP), in contrast to chemical phosphorylation modifications. HCPP can induce biomimetic mineralization itself without polymer additives by alternatively attracting calcium and phosphate through electrostatic attraction. Therefore, we herein propose the dual functions of amphiphilic MDP monomer with de- and remineralizing ability. MDP in the free state can demineralize dentin substrates by releasing protons, whereas MDP in the collagen-bound state as HCPP can induce intrafibrillar mineralization. The dual functions of MDP monomer with de- and remineralization properties might create a new epoch in adhesive dentistry and preventive dentistry.

Dynamic contract-enhanced CT-based radiomics for differentiation of pancreatobiliary-type and intestinal-type periampullary carcinomas.

AIM: To investigate whether computed tomography (CT) radiomics can differentiate pancreatobiliary-type from intestinal-type periampullary carcinomas. MATERIALS AND METHODS: CT radiomics of 96 patients (54 pancreatobiliary type and 42 intestinal type) with surgically confirmed periampullary carcinoma were assessed retrospectively. Volumes of interest (VOIs) were delineated manually. Radiomic features were extracted from preoperative CT images. A single-phase model and combined-phase model were constructed. Five-fold cross-validation and five machine-learning algorithms were utilised for model construction. The diagnostic performance of the models was evaluated by receiver operating characteristic (ROC) curves, and indicators included area under the curve (AUC), accuracy, sensitivity, specificity, and precision. ROC curves were compared using DeLong's test. RESULTS: A total of 788 features were extracted on each phase. After feature selection using least absolute shrinkage and selection operator (LASSO) algorithm, the number of selected optimal feature was 18 (plain scan), nine (arterial phase), two (venous phase), 23 (delayed phase), 15 (three enhanced phases), and 29 (all phases), respectively. For the single-phase model, the delayed-phase model using the logistic regression (LR) algorithm showed the best prediction performance with AUC, accuracy, sensitivity, specificity, and precision of 0.89, 0.83, 0.80, 0.88, and 0.93, respectively. Two combined-phase models showed better results than the single-phase models. The model of all phases using the LR algorithm showed the best prediction performance with AUC, accuracy, sensitivity, specificity, and precision of 0.96, 0.88, 0.90, 0.93, and 0.92, respectively. CONCLUSION: Radiomic models based on preoperative CT images can differentiate pancreatobiliary-type from intestinal-type periampullary carcinomas, in particular, the model of all phases using the LR algorithm.

SNHG16 promotes cell proliferation and migration through sponging miR-132 in melanoma.

Melanoma, which originates from the transformation of normal melanocytes, is one of the three main types of skin cancer. We aimed to explore the functions of SNHG16 and miR-132 in melanoma. CCK-8, Transwell assays were used to measure the viability and migration, respectively. Spearman's correlation analysis was performed to analyze the relationship between the expression of SNHG16, miR-132 and LAPTM4B in melanoma tissues. SNHG16 was overexpressed, and miR-132 was low expressed in melanoma tissues and cell lines. Moreover, overexpression of SNHG16 was associated with poor prognosis of melanoma patients. The expression of SNHG16 had a negative connection with the expression of miR-132, and it had a positive relationship with the expression of LAPTM4B in melanoma tissues. Knockdown of SNHG16 or overexpression of miR-132 inhibited SK-MEL-2 cell proliferation and migration. In addition, we confirmed that SNHG16 directly binding to miR-132 promotes the expression of LAPTM4B, facilitating the tumorigenesis of melanoma. SNHG16 promotes the expression of LAPTM4B by sponging miR-132, thereby acting as an oncogene in melanoma. This study demonstrated that the lncRNA-miRNA-mRNA signal cascade existed in melanoma, which may help elucidate the tumorigenesis and development mechanism of melanoma.

Circular RNA hsa_circ_0011946 promotes cell growth, migration, and invasion of oral squamous cell carcinoma by upregulating PCNA.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Circular RNA hsa_circ_0011946 promotes cell growth, migration, and invasion of oral squamous cell carcinoma by upregulating PCNA, by Y. Meng, E.-Y. Zhao, Y. Zhou, D.-X. Qiang, S. Wang, L. Shi, L.-Y. Jiang, L.-J. Bi, published in Eur Rev Med Pharmacol Sci 2020; 24 (3): 1226-1232-DOI: 10.26355/eurrev_202002_20175-PMID: 32096152" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20175.

Circular RNA hsa_circ_0011946 promotes cell growth, migration, and invasion of oral squamous cell carcinoma by upregulating PCNA.

OBJECTIVE: The importance of circular RNAs in malignant tumors has been well concerned nowadays. Oral squamous cell carcinoma (OSCC) is diagnosed prevalently in the world. Our study aims to uncover the potential functions of hsa_circ_0011946 in OSCC development. PATIENTS AND METHODS: Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was performed to determine the level of hsa_circ_0011946 in OSCC tissues and cell lines. Hsa_circ_0011946 was knocked down in OSCC cells. Biological functions of hsa_circ_0011946 in OSCC were identified by performing cell proliferation assay, colony formation assay, wound healing assay, and transwell assay. The underlying mechanism of hsa_circ_0011946 in regulating OSCC progression was explored by RT-qPCR and Western blot assay. RESULTS: Hsa_circ_0011946 was highly expressed in OSCC tissues compared with adjacent samples. It was also upregulated in OSCC cell lines. The knockdown of hsa_circ_0011946 inhibited cell growth, migration, and invasion in OSCC. The expression of PCNA was reduced via knockdown of hsa_circ_0011946. Furthermore, the expression of PCNA in tumor tissues was positively correlated to the expression of hsa_circ_0011946. CONCLUSIONS: Hsa_circ_0011946 could promote cell growth, migration, and invasion of OSCC by upregulating PCNA, which may offer a new therapeutic intervention for OSCC patients.

A comparison of open, laparoscopic and robotic total mesorectal excision: trial sequential analysis and network meta-analysis.

AIM: Total mesorectal excision (TME) for rectal cancer can be achieved by employing open (OpTME), laparoscopic (LaTME) and robotic (RoTME) approaches but which of these has the best outcome? The aim of present study is to identify the most effective technique for rectal cancer by comparing all outcomes. METHODS: Randomized controlled trials (RCTs) which compared at least two TME strategies were identified by literature search of electronic databases of articles published to June 2018. Network meta-analysis with trial sequential analysis was performed using a frequentist approach with random-effects meta-analysis. Data collection and analysis We conducted a systematic search of PubMed, EmBase, the Cochrane Library, CNKI, and Web of Science. Titles and abstracts of the retrieved publications were independently and blindly assessed by two authors. RESULTS: Twenty-two RCTs with 4882 rectal cancer patients were included in this analysis. The trial sequential analysis demonstrated that the cumulative Z-curve crossed either the traditional boundary or the trial sequential monitoring boundaries, suggesting that OpTME resulted in a more complete TME specimen than LaTME (relative risk 1.05, 95% confidence interval 1.01-1.08). Network meta-analysis showed there was no significant difference in the other comparisons. Based on the P score of completeness of the TME specimen and circumferential resection margin positivity, the best technique was OpTME, followed by RoTME and then LaTME. However, this order was reversed when complications and mortality were considered. RoTME led to better lymph node harvest. CONCLUSIONS: Although OpTME may give better pathological specimens, minimally invasive techniques may have advantages when considering lymph node harvest, complications and mortality. More RCTs are needed to determine which technique actually gives the best chance of survival.

Effects of dexmedetomidine on sepsis-induced liver injury in rats.

OBJECTIVE: To explore the effect of dexmedetomidine (DEX) on sepsis-induced liver injury in rats and the mechanism of action, providing certain references for the prevention and treatment of sepsis-induced liver injury in clinical practice. MATERIALS AND METHODS: A total of 60 male Sprague Dawley (SD) rats were randomly divided into 3 groups, namely sham operation group (Sham group, n=20), sepsis-induced liver injury group [lipopolysaccharides (LPS) group, n=20], and sepsis-induced liver injury + DEX group (LPS + DEX group, n=20) using a random number table. Rat models of sepsis-induced liver injury were established by intraperitoneal injection of LPS (10 mg/kg), and at the same time, DEX was intragastrically injected at a dose of 50 µg/kg. After 24 h, the survival analysis curves of each group of rats were plotted. Meanwhile, the levels of liver function indexes and oxidative stress markers were measured at 12 h in each group of rats. Hematoxylin-eosin (H&E) staining assay was carried out to detect the morphological changes of rat liver cells in each group. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL) staining assay was performed to detect the apoptosis level in rat liver tissues in each group. In addition, the expression level of Caspase 3 in three groups of rats was measured through immunohistochemical staining assay. Lastly, the effect of DEX on the protein expression of extracellular signal-regulated kinases 1/2 (ERK1/2) in liver tissues was detected via Western blotting. RESULTS: DEX significantly improved liver dysfunction induced by LPS and raised the 24 h-survival rates of rats (p<0.05). Besides, H&E staining results showed that DEX clearly relieved the pathological damage of rat liver cells caused by LPS. In comparison with LPS group, LPS + DEX group displayed more neatly arranged liver cells, less degradation and necrosis, and evidently attenuated cellular edema. Immunohistochemistry results revealed that DEX significantly reversed the increase in Caspase 3 expression resulting from LPS. The results of the TUNEL staining assay showed that DEX clearly inhibited the apoptosis of rat liver cells induced by LPS. The results of Western blotting revealed that DEX notably reversed the decrease of phosphorylated ERK1/2 (p-ERK1/2) in rat liver tissues compared with LPS group. CONCLUSIONS: DEX is able to markedly relieve LPS-induced liver injury in rats and the underlying mechanism may be related to the activation of the ERK1/2 signaling pathway.

A randomized controlled trial of oral chloral hydrate vs intranasal dexmedetomidine plus buccal midazolam for auditory brainstem response testing in children.

BACKGROUND: Moderate to deep sedation is required for an auditory brainstem response test when high-intensity stimulation is used. Chloral hydrate is the most commonly used sedative, whereas intranasal dexmedetomidine is increasingly used in pediatric non-painful procedural sedations. OBJECTIVE: The aim of this study was to compare the sedation success rate after oral chloral hydrate at 50 mg kg-1 and intranasal dexmedetomidine at 3 µg kg-1 plus buccal midazolam at 0.1 mg kg-1 for an auditory brainstem response test. METHODS: Children who required an auditory brainstem response test were recruited and randomly assigned to receive oral chloral hydrate at 50 mg kg-1 and intranasal placebo, or intranasal dexmedetomidine at 3 µg kg-1 with buccal midazolam 0.1 mg kg-1 . The primary outcome was the rate of successful sedation for auditory brainstem response tests. RESULTS: Fifty-seven out of 82 (69.5%) were successfully sedated after chloral hydrate, while 70 out of 78 (89.7%) children were successfully sedated with dexmedetomidine plus midazolam combination, with the odd ratio (95% CI) for successful sedation between dexmedetomidine plus midazolam combination and chloral hydrate estimated to be 3.84 (1.61-9.16), P = 0.002. Dexmedetomidine plus midazolam was associated with quicker onset with median onset time 15 (IQR 11.0-19.8) for dexmedetomidine plus midazolam and 20 (IQR 15.0-27.0) for chloral hydrate respectively, with difference between median (95% CI) of 5 [3-8], P < 0.0001). The behavior observed during drug administration of intranasal dexmedetomidine and buccal midazolam was better that of the children who had oral chloral hydrate. No children required oxygen therapy or medical intervention for hemodynamic disturbances in this study and the incidence of hypotension and bradycardia was similar. CONCLUSION: Intranasal dexmedetomidine plus buccal midazolam was associated with higher sedation success with deeper level of sedation, with similar discharge time and adverse event rate when compared to chloral hydrate.

LncRNA MT1JP acts as a tumor inhibitor via reciprocally regulating Wnt/ß-Catenin pathway in retinoblastoma.

OBJECTIVE: The aim of this study was to investigate the roles of MT1JP and ß-catenin in retinoblastoma. PATIENTS AND METHODS: We performed quantitative Reverse Transcriptase-Polymerase Chain Reaction (qRT-PCR) to quantify the expressions of MT1JP and ß-catenin in 44 retinoblastoma tissues and matched non-tumor tissues. What's more, retinoblastoma cell lines were transfected with pcDNA3.1-MT1JP, after which proliferation, cell cycle, apoptosis, and expression of ß-catenin as well as its downstream targets were assayed. We also conducted TOP-Flash reporter assay to explore the activity of Wnt/ß-catenin signaling pathway. RESULTS: The results revealed that MT1JP was down-regulated, while ß-catenin was highly expressed in retinoblastoma cells. Meanwhile, the forced expression of MT1JP impaired the expression of the ß-catenin protein and its downstream targets such as cyclin D1, c-myc. CONCLUSIONS: We demonstrated that MT1JP was a tumor suppressor by negatively modulating the activity of the Wnt/ß-catenin signaling pathway in the development of retinoblastoma and might function as a prognostic biomarker and therapeutic target.

The prevalence and influencing factors for child neglect in a rural area of Anhui province: a 2-year follow-up study.

OBJECTIVES: The purpose of the current study was to identify the change of prevalence and influencing factors for child neglect in a rural area of Anhui province through the 2-year follow-up study. STUDY DESIGN: Longitudinal study with 2-year follow-up. METHODS: Analyses were based on data from a longitudinal study, performed in five elementary schools and three secondary schools in Changfeng County. A total of 816 children aged between 7 and 16 years completed the three assessments during the period of 2009-2011. Generalized estimating equations (GEEs) were applied to identify the influencing factors of child neglect. RESULTS: The prevalence of child neglect was 67.8%, 56.6%, and 57.7% at the three assessments, respectively. There were 272 children (33.3%) having consistently experiencing neglect during three assessments and 106 (13.0%) children had not suffered from neglect during three assessments. Among 553 participants who experienced neglect at the first assessment, 105 (19.0%) children no longer met the diagnosis at the next two assessments. Fifty-two children who did not suffer from neglect at the first assessment experienced neglect at the final assessment. The results of GEEs showed that child neglect was clearly associated with age (odds ratio [OR] = 0.95, 95% confidence interval [CI] = 0.92-0.99, P = 0.016), male gender (OR = 1.20, 95% CI = 1.00-1.43, P = 0.047), siblings (OR = 1.26, 95% CI = 1.03-1.55, P = 0.028), parental marital disruption (OR = 2.02, 95% CI = 1.09-3.78, P = 0.027), left-behind status (OR = 1.26, 95% CI = 1.06-1.49, P = 0.008), severe family dysfunction (OR = 1.46, 95% CI = 1.03-2.07, P = 0.035), quality of life (OR = 0.98, 95% CI = 0.98-0.99, P < 0.001), positive coping styles (OR = 0.97, 95% CI = 0.94-0.99, P = 0.001), and negative coping styles (OR = 1.03, 95% CI = 1.02-1.05, P < 0.001). CONCLUSION: Our studies detected the decreased prevalence of child neglect across the three assessments. Additionally, some sociodemographic, psychosocial and family risk factors of child neglect were identified, which will be helpful for child neglect prevention strategies development and implementation in China.

[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.

Microbial sensor for drug susceptibility testing of Mycobacterium tuberculosis.

AIMS: Drug susceptibility testing (DST) of clinical isolates of Mycobacterium tuberculosis is critical in treating tuberculosis. We demonstrate the possibility of using a microbial sensor to perform DST of M. tuberculosis and shorten the time required for DST. METHODS AND RESULTS: The sensor is made of an oxygen electrode with M. tuberculosis cells attached to its surface. This sensor monitors the residual oxygen consumption of M. tuberculosis cells after treatment with anti-TB drugs with glycerine as a carbon source. In principle, after drug pretreatment for 4-5 days, the response differences between the sensors made of drug-sensitive isolates are distinguishable from the sensors made of drug-resistant isolates. The susceptibility of the M. tuberculosis H37Ra strain, its mutants and 35 clinical isolates to six common anti-TB drugs: rifampicin, isoniazid, streptomycin, ethambutol, levofloxacin and para-aminosalicylic acid were tested using the proposed method. The results agreed well with the gold standard method (LJ) and were determined in significantly less time. The whole procedure takes approximately 11 days and therefore has the potential to inform clinical decisions. CONCLUSIONS: To our knowledge, this is the first study that demonstrates the possible application of a dissolved oxygen electrode-based microbial sensor in M. tuberculosis drug resistance testing. This study used the microbial sensor to perform DST of M. tuberculosis and shorten the time required for DST. SIGNIFICANCE AND IMPACT OF THE STUDY: The overall detection result of the microbial sensor agreed well with that of the conventional LJ proportion method and takes less time than the existing phenotypic methods. In future studies, we will build an O2 electrode array microbial sensor reactor to enable a high-throughput drug resistance analysis.

[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 .