Magnetic covalent organic frameworks for extraction and determination of endocrine-disrupting chemicals in beverage and water samples.

BACKGROUND: Phenolic endocrine-disrupting chemicals (EDCs) are widespread and easily ingested through the food chain. They pose a serious threat to human health. Magnetic solid-phase extraction (MSPE) is an effective sample pre-treatment technology to determine traces of phenolic EDCs. RESULTS: Magnetic covalent organic framework (COF) (Fe3 O4 @COF) nanospheres were prepared and characterized. The efficient and selective extraction of phenolic EDCs relies on a large specific surface and the inherent porosity of COFs and hydrogen bonding, π-π, and hydrophobic interactions between COF shells and phenolic EDCs. Under optimal conditions, the proposed magnetic solid-phase extraction-high-performance liquid chromatography-ultra violet (MSPE-HPLC-UV) based on the metallic covalent organic framework method for phenolic EDCs shows good linearities (0.002-6 µg mL-1 ), with R2 of 0.995 or higher, and low limits of detection (6-1.200 ng mL-1 ). CONCLUSION: Magnetic covalent organic frameworks (Fe3 O4 @COFs) with good MSPE performance for phenolic EDCs were synthesized by the solvothermal method. The magnetic covalent organic framework-based MSPE-HPLC-UV method was applied successfully to determine phenolic EDCs in beverage and water samples with satisfactory recoveries (90.200%-123%) and relative standard deviations (2.100%-12.100%). © 2023 Society of Chemical Industry.

A novel cell-wall polysaccharide derived from the stipe of Agaricus bisporus inhibits mouse melanoma proliferation and metastasis.

Malignant melanoma is an invasive and highly aggressive skin cancer that-if diagnosed-poses a serious threat to the patient's health and life. In this work, a novel purified cell-wall polysaccharide (termed Abwp) was obtained from the discarded stipe of Agaricus bisporus (A. bisporus) and characterized to be a novel homogeneous polysaccharide consisted of a ß-(1 â†’ 4)- glucosyl backbone with ß-(1 â†’ 2) and (1 â†’ 6)-d-glucosyl side-chains. The anti-melanoma effects of Abwp and its associated mechanisms in mice were then explored using in vitro and in vivo approaches. In vitro results showed that Abwp inhibited B16 melanoma cell proliferation and promoted their apoptosis in both time- and dose-dependent manners. In B16 cells induced with tumor necrosis factor (TNF-α), Abwp significantly decreased the protein expression of inflammatory-related signaling pathway (e.g., p38 MAPK and NF-κB) in time-, concentration-, and dose-dependent manners. Moreover, Abwp blocked nuclear entry of NF-κB-p65. In an in vivo mouse model featuring neoplasm transplantation with B16 melanoma cells, Abwp significantly inhibited the growth and proliferation of mouse melanoma. Hematoxylin staining showed that the invasion of melanoma cells into the lung tissue of the Abwp-treated group was significantly reduced. Immunohistochemical analysis showed that the expression of proliferation cell nuclear antigen (PCNA), N-cadherin, MMP-9, and Snail in the lung of mouse was significantly inhibited. Immunofluorescence showed that Abwp significantly interfered with the nuclear transcription of NF-κB-p65 in a dose-dependent manner. Collectively, these results showed that Abwp mediated p38 MAPK and NF-κB signaling pathways to inhibit the inflammatory response and malignant proliferation and metastasis of melanoma in mice.

Clinical application of ultrasound-guided percutaneous microwave ablation for benign breast lesions: a prospective study.

BACKGROUND: Background: Benign breast lesions are the most common diseases in adult women, which have been treated with minimally invasive therapies in recent years. Little is known about the feasibility of Microwave ablation (MWA) for benign breast lesion treatment. The primary aim of this prospective study was to evaluate the safety and efficiency of MWA as a potential therapeutic option for benign breast lesions in a single-center cohort study. METHODS: Women with possibly benign breast lesions based on an ultrasound (US) assessment who were scheduled to undergo MWA between November 2014 to July 2018 were included in the study. The patients underwent conventional US to measure the size of the lesion, Doppler US to assess the vascularity of the lesion, elastography to evaluate the stiffness of the mass, core needle biopsy of suspicious lesions, contrast-enhanced US to help determine the treatment plan and eventually MWA of the lesion. Lesions were followed at one, three, six, twelve and eighteen months after treatment to with the same imaging modalities. RESULTS: A total of 314 women aged 17 to 69 years old (mean = 36.9 ± 9.9 years) with 725 benign breast lesions (mean of maximum diameter = 10.86 ± 5.40 mm) were included. The frequency of palpable mass, pain and nipple discharge significantly decreased after treatment. Complete ablation rate was 97.8%, immediately after ablation, which increased to 100% after supplementary ablation of the 15 cases with incomplete ablation. Blood flow classification and lesion's volume also showed a significant decrease, while both volume reduction ratio and disappearance rate significantly increased following treatment. The elasticity score of the lesions showed fluctuations across different follow-up intervals. None of the patients experienced major complications and the 1% who had mild symptoms were successfully treated. CONCLUSION: MWA treatment is shown to be safe and efficient and has the potential to be considered as an alternative first line treatment for benign breast lesions.

Prediction of kidney transplant outcome based on different DGF definitions in Chinese deceased donation.

BACKGROUND: Delayed graft function (DGF) is an important complication of kidney transplantation and can be diagnosed according to different definitions. DGF has been suggested to be associated with the long-term outcome of kidney transplantation surgery. However, the best DGF definition for predicting renal transplant outcomes in Chinese donations after cardiac death (DCDs) remains to be determined. METHOD: A total of 372 DCD kidney transplant recipients from June 2013 to July 2017 in the First Affiliated Hospital of Xi'an Jiaotong University were included in this retrospective study to compare 6 different DGF definitions. The relationships of the DGF definitions with transplant outcome were analyzed, including graft loss (GL) and death-censored graft loss (death-censored GL). Renal function indicators, including one-year estimated glomerular filtration rate (eGFR) and three-year eGFR, and were compared between different DGF groups. RESULTS: The incidence of DGF varied from 4.19 to 35.22% according to the different DGF diagnoses. All DGF definitions were significantly associated with three-year GL as well as death-censored GL. DGF based on requirement of hemodialysis within the first week had the best predictive value for GL (AUC 0.77), and DGF based on sCr variation during the first 3 days post-transplant had the best predictive value for three-year death-censored GL (AUC 0.79). Combination of the 48-h sCr reduction ratio and classical DGF can improve the AUC for GL (AUC 0.85) as well as the predictive accuracy for death-censored GL (83.3%). CONCLUSION: DGF was an independent risk factor for poor transplant outcome. The combination of need for hemodialysis within the first week and the 48-h serum creatinine reduction rate has a better predictive value for patient and poor graft outcome.

IL-18 genetic polymorphisms may contribute to the pathogenesis of tuberculosis among Asians: a meta-analysis of case-control studies.

This current meta-analysis of case-control studies was continued to investigate whether the genetic polymorphisms of IL-18 gene contribute to the occurrence and progression of tuberculosis (TB). We searched certain English and Chinese databases for relevant studies without language restrictions. Meta-analysis for the moment was performed with the adoption of the STATA statistical software. Crude OR and its corresponding 95 % confidence interval (95 % CI) were calculated as estimates of relative risk for UC under different genetic models. Seven case-control studies (TB patients = 1,325, healthy subjects = 1,778) were included for the following analysis. We evaluated two functional polymorphisms (rs1946518 C>A and rs187238 G>C). Pooled OR within the progression of statistical analysis indicated that the specific polymorphism of IL-18 rs1946518 C>A showed a closely relationship with the elevated susceptibility to TB under those three genetic models (allele model: OR 1.24, 95 % CI 1.11-1.38, P < 0.001; dominant model: OR 1.41, 95 % CI 1.21-1.65, P < 0.001; homozygous model: OR 1.46, 95 % CI 1.15-1.86, P = 0.002; respectively). However, we observed no statistical associations of the IL-18 rs187238 G>C polymorphism with the susceptibility to TB under any of the genetic models (all P > 0.05). Country-stratified analysis results detected that the variants of IL-18 may be strongly enrolled in the risk of TB among populations in China (allele model: OR 1.19, 95 % CI 1.06-1.33, P = 0.003; recessive model: OR 1.54, 95 % CI 1.00-2.36, P = 0.048; homozygous model: OR 1.59, 95 % CI 1.09-2.33, P = 0.016; respectively), but not among populations in Iran, Korea and India (all P > 0.05). Current results provide strong evidence that IL-18 mutations may be evidently related to the occurrence and development of TB, especially for the rs1946518 C>A polymorphism among populations in China.

Relationships of COX2 and MMP12 genetic polymorphisms with chronic obstructive pulmonary disease risk: a meta-analysis.

We performed the present meta-analysis in an attempt to confirm the correlation of genetic polymorphisms in the COX2 and MMP12 genes with the susceptibility to chronic obstructive pulmonary disease (COPD). We searched English database such as PubMed, CISCOM, CINAHL, Web of Science, Google Scholar and several Chinese database for meta-analysis. There were no specific language restrictions. Two investigators systematically extracted relevant data within those included studies. Crude ORs with its corresponding 95 % CI were calculated. STATA 12.0 software was adopted for statistical analysis. The impact of COX2 and MMP12 genetic polymorphisms on the pathogenesis of COPD was investigated in the current study with a total of 10 case-control studies, which includes 1,751 COPD patients and 2,472 healthy subjects. Four common polymorphisms, including rs689466 G > A and rs20417 G > C in the COX2 gene, rs652438 A > G and rs2276109 A > G were evaluated in the MMP12 gene. Pooled OR of the present studies and results showed that the frequency of COX2 rs20417 polymorphism was prevalent in COPD patients than those of healthy subjects (C allele vs. G allele OR = 1.33, 95 % CI 1.06-1.67, P = 0.014; GC + CC vs. GG OR = 1.86, 95 % CI 1.07-3.24, P = 0.029; respectively). However, we found no significant correlation between COX2 rs689466 polymorphism and the risk of COPD (all P > 0.05). Furthermore, our meta-analysis illustrated that individuals with MMP12 rs652438 polymorphism had significantly increased risk of developing COPD (G allele vs. A allele OR = 1.62, 95 % CI 1.08-2.42, P = 0.020; AG + GG vs. AA OR = 2.14, 95 % CI 1.12-4.09, P = 0.021; respectively). Nevertheless, no positive relation was detected between MMP12 rs2276109 variant and the risk of COPD. Our meta-analysis indicates that COX2 and MMP12 genetic polymorphisms may be strongly implicated in the development of COPD, especially for the COX2 rs20417 and MMP12 rs652438 polymorphisms. Thus, COX2 and MMP12 genetic polymorphisms could potentially be utilized as helpful biomarkers for early diagnosis of COPD.

[Evaluation of immune status of kidney transplant recipients by combined HLA-G5 and sCD30].

OBJECTIVE: to study the relationship between the expression of serum human leucocyte antigen-G5 (HLA-G5)/soluble CD30 (sCD30) and the function of renal graft in kidney transplant recipients and investigate the immune status of recipients with combined HLA-G5 and sCD30. METHODS: from January 2002 to November 2008, a total of 66 kidney transplant recipients in our centre were selected as subjects and divided into three groups: stable function of renal graft (n = 38), acute rejection (n = 15) and chronic rejection (n = 13). The expressions of serum HLA-G5 and sCD30 were detected. There were two different immune conditions with acute/chronic allograft rejection and normal renal graft in kidney transplant recipients as evaluated by combined HLA-G5 and sCD30. The sensitivity, specificity and critical value of the method were analyzed by the curve of receiver operating characteristic. RESULTS: the levels of HLA-G5 and sCD30 were significantly correlated with serum creatinine (r = -0.493, 0.691, both P < 0.01). Within the first year post-transplantation, the sensitivity was 78.6% and the specificity 85.7% when HLA-G5 critical value 82 microg/L and sCD30 critical value 12.2 microg/L. After one year post-transplantation: the sensitivity was 92.3% and the specificity 84.6% when HLA-G5 critical value 141 microg/L and sCD30 critical value 10.3 microg/L. CONCLUSION: the immune state of recipients are evaluated by combine HLA-G5 and sCD30 which may be a simple and valid method.

Repairing and Analgesic Effects of Umbilical Cord Mesenchymal Stem Cell Transplantation in Mice with Spinal Cord Injury.

Transplantation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into spinal cord injury (SCI) may alleviate neuropathic pain and promote functional recovery. The underlying mechanism likely involves activation of glial cells and regulation of inflammatory factors but requires further validation. SCI was induced in 16 ICR mice using an SCI compression model, followed by injection of lentiviral vector-mediated green fluorescent protein- (GFP-) labeled hUC-MSCs 1 week later. Behavioral tests, histological evaluation, and inflammatory factor detection were performed in the treatment (SCI+hUC-MSCs) and model (SCI) groups. Histological evaluation revealed GFP expression in the spinal cord tissue of the treatment group, implying that the injected MSCs successfully migrated to the SCI. The Basso, Beattie, and Bresnahan (BBB) scores showed that motor function gradually recovered over time in both groups, but recovery speed was significantly higher in the treatment group than in the model group. The pain threshold in mice decreased after SCI but gradually increased over time owing to the self-repair function of the body. The corresponding pain threshold of the treatment group was significantly higher than that of the model group, indicating the therapeutic and analgesic effects of hUC-MSCs. Expression of IL-6 and TNF-α in the spinal cord tissue of the treated group decreased, whereas glial cell line-derived neurotrophic factor (GDNF) expression along with ED1 expression increased compared with those in the model group, suggesting that SCI activated ED1 inflammatory macrophages/microglia, which were subsequently reduced by hUC-MSC transplantation. hUC-MSCs are speculated to enhance the repair of the injured spinal cord tissue and exert an analgesic effect by reducing the secretion of inflammatory factors IL-6 and TNF-α and upregulating the expression of GDNF.

Early Immunosuppressive Exposure of Enteric-Coated-Mycophenolate Sodium Plus Tacrolimus Associated with Acute Rejection in Expanded Criteria Donor Kidney Transplantation.

BACKGROUND: Immunosuppressive agents are still inefficient in preventing biopsy-proven acute rejection (BPAR) after expanded criteria donor (ECD) kidney transplantation. The aim of this study was to investigate the relationships between early immunosuppressive exposure and the development of BPAR. METHODS: We performed a retrospective study of 58 recipients of ECD kidney transplantation treated with enteric-coated-mycophenolate sodium, tacrolimus (Tac), and prednisone. The levels of mycophenolic acid-area under the curve (MPA-AUC)0-12h and Tac C0were measured at the 1st week and the 1st month posttransplant, respectively. The correlation was assessed by multivariate logistic regression. RESULTS: The occurrence rates of BPAR and antibody-mediated rejection were 24.1% and 10.3%, respectively. A low level of MPA-AUC0-12h at the 1st week posttransplant was found in BPAR recipients (38.42 ± 8.37 vs. 50.64 ± 13.22, P < 0.01). In addition, the incidence of BPAR was significantly high (P < 0.05) when the MPA-AUC0-12hlevel was <30 mg·h-1·L-1 at the 1st week (15.0% vs. 44.4%) or the Tac C0was <4 ng/ml at the 1st month posttransplant (33.3% vs. 21.6%). Multivariable logistic regression analysis showed that the MPA-AUC0-12h at the 1st week (OR: 0.842, 95% CI: 0.784-0.903) and the Tac C0at the 1st month (OR: 0.904, 95% CI: 0.822-0.986) had significant inverse correlation with BPAR (P < 0.05). CONCLUSIONS: Low-level exposure of MPA and Tac C0in the early weeks posttransplant reflects an increased acute rejection risk, which suggested that MPA-AUC0-12h <30 mg·h-1·L-1 and Tac C0 <4 ng/ml should be avoided in the first few weeks after transplantation.

Effects of chronic unpredictable mild stress on ovarian reserve in female rats: Feasibility analysis of a rat model of premature ovarian failure.

Premature ovarian failure (POF) results from a number of disorders. The POF model is primarily based on chemotherapeutic injury, and hence is not suitable for assessing the effects of chronic stress on ovarian function. Therefore, improved animal models are required to analyze the effects of chronic stress on ovarian reserve. The feasibility of the chronic unpredictable mild stress (CUMS) method for establishing a model of POF was examined. The depressive behavior exhibited by rats was evaluated with the open field and sucrose preference tests. Vaginal smears were obtained for assessment of the estrous cycle. The ovarian reserve of the animals was evaluated using the estrous cycle, ovarian histology and serum levels of gonadotropin releasing hormone (GnRH), follicle­stimulating hormone (FSH), estradiol (E2), and anti­Müllerian hormone (AMH). Compared with the control group, body weight, time spent in the center, horizontal movement, vertical frequency, consumption of sucrose, sucrose preference, number of small follicles from the rats, and serum E2, AMH and GnRH levels were significantly decreased in the CUMS group (all P<0.05). However, the estrous cycle was prolonged significantly (P<0.05) and serum FSH levels were increased significantly (P<0.01). These results suggested that the CUMS model rats exhibited depression­like behaviors. CUMS may induce psychological stress and decrease ovarian reserve in female rats. Thus, the CUMS model may be used to assess the effects of chronic stress on female reproductive function.

[Impacts of kidney transplantation on erectile function and its contributing factors evaluation].

OBJECTIVE: To study the impacts of kidney transplantation on erectile function and analyse its contributing factors. METHODS: In order to evaluate the severity of erectile dysfunction (ED), a total of 250 married male kidney transplant recipients (KTR) with functioning graft were assessed with the International Index of Erectile Function (IIEF) questionnaire. Data of clinical characteristics, medical and sexual history and laboratory examination were collected. Univariate and multivariate logistic regression analyses were carried out to determine which have independent impacts on erectile function. RESULTS: The investigation was accomplished in 84.8% of the KTRs. There was no significant difference in ED incidence before and after renal transplantation (53.8% vs. 44.3%, P > 0.05). According to the IIEF score, erectile function improved in 43.9% of the KTRs, remained unchanged in 42.9%, and deteriorated in 13.2%, as compared with pre-transplantation. Logistic regression analysis showed that significant and independent influencing factors in erectile function were age, hemoglobin level, presence of DM and/or peripheral neuropathy and iterative transplantations, and their relative risks were 3.01, 2.01, 3.15, 3.89 and 2.67, respectively. CONCLUSION: ED is highly prevalent among KTRs and its pathogenesis is multifactorial. Age, presence of DM and/or peripheral neuropathy, hemoglobin level and iterative transplantations were chief contributing factors in erectile function.

Ultrasound-guided percutaneous microwave ablation for benign breast lesions: evaluated by contrast-enhanced ultrasound combined with magnetic resonance imaging.

BACKGROUND: Minimally invasive ablative techniques in the treatment of breast tumor has become popularly in recent years. METHODS: We analyzed gray-scale and contrast-enhanced ultrasound (CEUS) features of 205 microwave ablated breast benign lesions from 182 consecutive patients, compared with magnetic resonance imaging (MRI) and histopathology findings. The follow-up was implemented at 3, 6 and 12 months after the ablation treatment. RESULTS: Before the MWA, the mean of largest diameter and volume of the lesions were 14.41±6.54 and 3,224±961 mm3, respectively. However, those of the lesions respectively were 8.48±6.30 and 2,116±732 mm3 one year after the treatment. The longest diameter and the volume of the ablative lesions were gradually decreased 3, 6 or 12 months after the MWA. 44 (/205, 21.5%) ablative lesions were disappeared one year after the MWA. One hundred and forty-two (/205, 69.3%) ablative lesions presented a hypoechoic halo surrounding it on gray-scale US after the MWA. The success rate of the MWA treatment in the benign breast lesion was 87.32% and 82.93% evaluated by CEUS and enhanced MRI, respectively. During the ablation, no patient had serious complications, such as hemorrhage, serious pain and fat necrosis, etc. CONCLUSIONS: Microwave ablation was a safe and efficient method in the treatment of the benign breast tumors. CEUS and enhanced MRI could accurately assess whether the MWA treatment is effective.

Predictive Score Model for Delayed Graft Function Based on Easily Available Variables before Kidney Donation after Cardiac Death.

BACKGROUND: How to evaluate the quality of donation after cardiac death (DCD) kidneys has become a critical problem in kidney transplantation in China. Hence, the aim of this study was to develop a simple donor risk score model to evaluate the quality of DCD kidneys before DCD. METHODS: A total of 543 qualified kidneys were randomized in a 2:1 manner to create the development and validation cohorts. The donor variables in the development cohort were considered as candidate univariate predictors of delayed graft function (DGF). Multivariate logistic regression was then used to identify independent predictors of DGF with P < 0.05. Date from validation cohort were used to validate the donor scoring model. RESULTS: Based on the odds ratios, eight identified variables were assigned a weighted integer; the sum of the integer was the total risk score for each kidney. The donor risk score, ranging from 0 to 28, demonstrated good discriminative power with a C-statistic of 0.790. Similar results were obtained from validation cohort with C-statistic of 0.783. Based on the obtained frequencies of DGF in relation to different risk scores, we formed four risk categories of increasing severity (scores 0-4, 5-9, 10-14, and 15-28). CONCLUSIONS: The scoring model might be a good noninvasive tool for assessing the quality of DCD kidneys before donation and potentially useful for physicians to make optimal decisions about donor organ offers.

Polyglycolic Acid Fibrous Scaffold Improving Endothelial Cell Coating and Vascularization of Islet.

BACKGROUND: Improving islet graft revascularization has become a crucial task for prolonging islet graft survival. Endothelial cells (ECs) are the basis of new microvessels in an isolated islet, and EC coating has been demonstrated to improve the vascularization and survival of an islet. However, the traditional method of EC coating of islets has low efficiency in vitro. This study was conducted to evaluate the effect of a polyglycolic acid (PGA) scaffold on the efficiency of islet coating by ECs and the angiogenesis in the coated islet graft. METHODS: A PGA fibrous scaffold was used for EC coating of islet culture and was evaluated for its efficiency of EC coating on islets and islet graft angiogenesis. RESULTS: In in vitro experiments, we found that apoptosis index of ECs-coating islet in PGA group (27% ± 8%) was significantly lower than that in control group (83% ± 20%, P < 0.05) after 7 days culture. Stimulation index was significantly greater in the PGA group than in the control group at day 7 after ECs-coating (2.07 ± 0.31 vs. 1.80 ± 0.23, P < 0.05). vascular endothelial growth factor (VEGF) level in the PGA group was significantly higher than the coating in the control group after 7 days culture (52.10 ± 13.50 ng/ml vs. 16.30 ± 8.10 ng/ml, P < 0.05). Because of a tight, circumvallated, adhesive and three-dimensional growth microenvironment, islet cultured in a PGA scaffold had higher coating efficiency showing stronger staining intensity of enzyme than those in the control group after 14 days of culture following ECs-coating. For in vivo study, PGA scaffold significantly prolonged the average survival time of EC-coated islet graft after transplantation compared with control group (15.30 ± 5.60 days vs. 8.30 ± 2.45 days, P < 0.05). The angiogenesis and area of survived grafts were more in the PGA group compared with the control group by measuring the mean microvessel density (8.60 ± 1.21/mm2 vs. 5.20 ± 0.87/mm2, P < 0.05). In addition, expression of VEGF and tyrosin-protein kinase receptor (Tie-2) gene increased in PGA scaffold group than that in control group by real-time reverse transcription-polymerase chain reaction analysis. CONCLUSIONS: These results demonstrate that the efficiency of EC coating of islets was successfully increased by culturing ECs on a PGA scaffold. This method enhances the function, survival, and vascularization of isolated islets in vitro and in vivo.

Treatment of cytomegalovirus infection after renal transplantation: experience from a single center in China.

BACKGROUND: We compared the efficacy and safety of 2 different treatments of CMV infection, including asymptomatic CMV replication and CMV disease. MATERIAL AND METHODS: 852 renal transplantation recipients, including asymptomatic CMV replication and CMV disease, received antiviral therapies of intravenous acyclovir or comprehensive anti-infection solution, mainly with intravenous ganciclovir. Effect, time, acute allograft rejection, and safety were analyzed during the antiviral therapy RESULTS: The total effective rates were higher with ganciclovir in both asymptomatic CMV replication (98.96% vs. 84.90%) and CMV disease (96.29% vs. 50.36%). Ganciclovir significantly shortened antiviral therapy duration in both asymptomatic CMV replication (15.0 ± 2.3 days vs. 16.0 ± 3.4 days) and CMV disease (19.7 ± 3.1 days vs. 21.5 ± 4.0 days). The acute allograft rejection incidences were significantly lower with ganciclovir in both asymptomatic CMV replication (8% vs. 14%) and CMV disease (11% vs. 22%). CMV-IEA was detected in renal grafts of patients with acute rejection. There was more CMV-associated acute rejection using acyclovir than using ganciclovir. Except for the higher incidence of anemia leucopenia and anemia with ganciclovir, the safety profiles of both drugs were similar. CONCLUSIONS: Comprehensive anti-infection solution, mainly with intravenous ganciclovir, can effectively treat CMV infection, shorten duration of therapy, and decrease acute rejection. The few adverse effects had negligible effects on use of ganciclovir.

[The effects of ICA on the cell-immunosuppressive and bone-marrow-suppressive mice after chemotherapy].

AIM: to observe the effect of ICA on the immunosuppressive and bone-marrow-suppressive mice after chemotherapy, and explore the machanism of hematopoietic and immunologic function in mice accentuated by ICA. METHODS: mice were injected Cy intraperitoneally except control group , then randomly divided into mode group(saline), positive (Shenqi, 1 mL/d)group, ICA groups(150, 80, 40 mg/kg.d). All mice were treated respectively for 10 successive days. The pathological changes of thymus were observed by HE staining. Killing activity of peritoneal macrophage were measured by LDH kits and its production of TNF-α and IL-12 was measured by ELISA kits. Population of white blood cells (WBC), red blood cell (RBC) and platelet (PLT) in the peripheral blood were detected with automated blood cell counter (ABCC). Hemogram of peripheral blood and bone marrow morphology were observed under the microscope. RESULTS: ICA could protect the thymus and bone marrow from damage. The proliferation of lymphocyte and killing activity of macrophage cells in ICA treatment groups was all enhanced, moreover, the population of WBC, RBC and PLT were increased significantly. CONCLUSION: ICA can improve the state of immunosuppressive and myelosuppressive mice caused by Cy thus could alleviate the side effect of chemotherapy effectively.

Allograft rejection-related gene expression in the endothelial cells of renal transplantation recipients after cytomegalovirus infection.

OBJECTIVE: To explore the effects of cytomegalovirus (CMV) infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. METHODS: Endothelial cells (ECs) were cultured and stimulated by a variety of factors: A, normal control group; B, inactivated human cytomegalovirus (HCMV) infection group; C, HCMV infection group; D, HCMV supernatant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 (ICAM-1) and major histocompability complex (MHC) class I and class II antigens was detected by flow cytometry (FCM) and immunohistochemistry. RESULTS: We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D (P>0.05). Although the expression levels of ICAM-1 were not significantly different between groups C and E (P>0.05), the ICAM-1 expression in these two groups was significantly higher than that in group A (P<0.05). ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E (P>0.05). Human leucocyte antigen (HLA)-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant differences of HLA-ABC and HLA-DR expression among groups A, B and D (P>0.05). However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported (P<0.05). Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C (P<0.05). CONCLUSION: CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection.

Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD4+ CD25+ Treg cells.

OBJECTIVE: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4(+) CD25(+) regulatory T (Treg) cells, in order to inform further studies in Treg cell function. METHODS: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4(+) CD25(+) Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4(+) CD25(+) Treg cells, trypan blue staining to detect cell viability, and propidium iodide (PI) staining to assess the cell viability. We detected the immune inhibition of CD4(+) CD25(+) Treg cells in the in vitro proliferation experiments. RESULTS: The results showed that compared to flow cytometry sorting, magnetic cell sorting took more time and effort, but fewer live cells were obtained than with flow cytometry sorting. The CD4(+) CD25(+) Treg cells, however, obtained with both methods have similar immunosuppressive capacities. CONCLUSION: The result suggests that both methods can be used in isolating CD4(+) CD25(+) Treg cells, and one can select the best method according to specific needs and availability of the methodologies.

[High-concentration palmitic acid inhibits the proliferation of peripheral blood-derived human endothelial progenitor cells in vitro].

OBJECTIVE: To study the effects of palmitic acid (PA) on the proliferation of peripheral blood-derived endothelial progenitor cells (EPCs) in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the peripheral blood by Ficoll density-gradient centrifugation. The isolated EPCs were characterized by Di-LDI uptake and FITC-lectin binding assay using laser confocal microscope, and further identified by detection of CD34, CD133 and VEGFR2 expression using flow cytometry. The cultured EPCs were incubated in the presence of PA at the concentrations of 0, 50, 100, 200, 400 and 800 micromol/L for different durations (0, 12, 24, 36, 48 and 60 h). The cell morphology was observed and cell proliferation determined with CCK-8 assay. RESULTS: Incubation with 400 and 800 micromol/L of PA significantly inhibited the proliferative ability of EPCs as compared with the control group (P < 0.05). PA at 400 micromol/L had the strongest effect on the cell proliferation, and this effect was intensified with the passage of time, reaching the peak at 48 h with the growth inhibition rate of 58.59% (P < 0.05). CONCLUSION: High-concentration PA can significantly inhibit the proliferation of EPCs in vitro.

[Protection of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by recombinant adenovirus containing hCGPx gene].

AIM: To study the protective effect of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by transfection of recombinant adenovirus-mediated human cytosolic glutathione peroxidase(hCGPx) gene. METHODS: Recombinant pGEM-T vector containing hCGPx cDNA and pACCMV-pLpA adenovirus shuttle plasmid was constructed. Then the shuttle plasmid pACCMV-hCGPx and pJM17 were co-transfected into 293 cells and recombinant adenovirus AdCMV-hCGPx was obtained. Cultured HK-2 cells were transfected with AdCMV-hCGPx or vacant recombinant adenovirus (control). The expression ratio of transfected hCGPx gene were studied. Cell viability, the percentage of apoptosis and death were evaluated after hypoxia-reoxygenation damage. RESULTS: The expression ratio of hCGPx gene was higher in the AdCMV-hCGPx transfected cells than that in the control group (P<0.01). After hypoxia-reoxygenation damage, the viability of hCGPx gene transfected cells was significantly higher than that of control and the percentage of apoptosis and death of hCGPx transfected cells was significantly lower than that of control. CONCLUSION: The transfection of hCGPx mediated by recombinant adenovirus could protect renal tubular epithelial cells from hypoxia-reoxygenation damage in vitro.