Clinical efficacy of ureteroscopy-assisted laparoscopic ureteroplasty in the treatment of ureteral stricture after pelvic surgery.

OBJECTIVE: This study is to investigate the safety and efficacy of ureteroscope-assisted laparoscopic ureteroplasty in treating ureteral stricture after pelvic surgery. METHODS: A retrospective analysis of the clinical data of 95 patients treated for ureteral stricture at Ganzhou People's Hospital from June 2017 to March 2023 after pelvic surgery. In this group, 49 patients underwent ureteroscope and laparoscopic ureteroplasty under lithotomy position. The control group consisted of 46 patients who underwent simple laparoscopic ureteroplasty in a supine position. Postoperative data from both groups were collected and compared, including operation time, amount of blood loss during surgery, postoperative hospital stay, incidence of complications, success rate of ureteroplasty, and effectiveness of the operation. RESULTS: The success rate of end-to-end ureteral anastomosis in the observation group was 93.88%, and the operation effectiveness rate was 100%. The success rate in the control group was 78.26% and the operation effectiveness rate was 89.1%.The average operation time and intraoperative blood loss in the observation group were (121.3 ± 44.6) min and (137.5 ± 34.2) ml, respectively, while in the control group they were (151.2 ± 52.3) min and (165.6 ± 45.8) ml, the difference were statistically significant (P < 0.05). The incidence of perioperative complications in the observation group was 2%, significantly lower than that in the control group (19.6%) (P < 0.05). CONCLUSION: Ureteroscope-assisted laparoscopic ureteroplasty for ureteral stricture after pelvic surgery has the advantages of shortened operation time, increased success rate, and reduced incidence of complications, making it an optional surgical scheme in clinical practice.

Clinical efficacy analysis of tip­flexible suctioning ureteral access sheath combined with disposable flexible ureteroscope to treat 2-4 cm renal stones.

PURPOSE: This study aims to evaluate the clinical efficacy of using a tip­flexible suctioning ureteral access sheath (TFS-UAS) in combination with a traditional ureteral access sheath (T-UAS) and a disposable flexible ureteroscope (DFU) for treating large renal stones (2-4 cm in diameter). METHODS: We retrospectively collected clinical data from 238 patients who underwent retrograde intrarenal surgery (RIRS) at Ganzhou People's Hospital between January 2019 and October 2023. The study included 238 patients who met the inclusion criteria, with 125 in the observation group using TFS-UAS and 113 in the control group using T-UAS. We compared differences in the stone-free rate (SFR), complication rates, surgery duration, and average hospital stay between the two groups. RESULTS: All 238 surgeries were successfully completed. The stone-free rates for the observation group at the first and thirtieth day post-surgery were 87.20% and 95.20%, respectively, whereas for the control group, the rates were 73.45% and 85.84%, showing statistically significant differences (P < 0.05). The overall complication rates were 1.6% for the observation group and 14.16% for the control group, also statistically significant (P < 0.001). The surgical times for stone removal were (101.17 ± 25.64) minutes for the observation group and (86.23 ± 20.35) minutes for the control group, with significant differences (P < 0.05). CONCLUSION: Compared to T-UAS, combining TFS-UAS with DFU for treating renal stones of 2-4 cm diameter, although more time-consuming, resulted in higher SFRs and improved safety.

Clinical efficacy analysis of partial cystectomy and radical cystectomy in the treatment of muscle-invasive sarcomatoid carcinoma of the urinary bladder.

Objective: This study compares the clinical efficacy of partial cystectomy (PC) versus radical cystectomy (RC) in the treatment of muscle-invasive bladder urothelial carcinoma (SCUB) through a retrospective analysis. Methods: We retrospectively analyzed the clinical data of 20 patients diagnosed with muscle-invasive SCUB from July 2015 to August 2023 at Ganzhou People's Hospital. All patients underwent surgical treatment followed by chemotherapy, with 9 receiving PC and 11 undergoing RC. We compared the average survival time of deceased patients for both treatments and conducted survival and multivariate analyses using the Kaplan-Meier method and Cox proportional hazards model, respectively. Results: All 20 patients were postoperatively diagnosed with muscle-invasive SCUB and were followed up for 4 to 60 months. The average survival time for patients undergoing PC was 11.5 months, with survival rates at 1 year, 2 years, and 5 years of 55.56%, 22.22%, and 11.11%, respectively. In contrast, patients receiving RC had an extended average survival time of 22.5 months, and their 1-year, 2-year, and 5-year survival rates increased to 63.64%, 36.36%, and 18.18%, respectively. Survival analysis revealed statistically significant differences in prognosis between PC and RC for the treatment of muscle-invasive SCUB (P<0.05). Conclusion: SCUB is a rare malignant tumor with unique biological characteristics often associated with poor prognosis. Upon diagnosis, RC should be considered as an early treatment approach when the patient's overall condition permits.

Dynamic Changes of Regulatory T Cells/CD4⁺ T Cells in Peripheral Blood of Adult Kidney Transplant Recipients: A Comparison of Pediatric and Adult Kidney Donors.

BACKGROUND Inducing transplantation tolerance and monitoring the recipient's immune status to improve allograft survival remains the main goal for kidney transplantation (KTx). MATERIAL AND METHODS A total of 53 renal transplantation patients and 20 healthy individuals were assigned to the post-transplantation and healthy groups, respectively; 10 recipients with stable renal function for 2 years after kidney transplantation were assigned to Group C. Eleven kidney transplantation recipients were hospitalized due to lung infection. Flow cytometry was used to measure levels of Tregs/CD4⁺ T cells. RESULTS The Tregs/CD4⁺ T cells ratio reached homeostasis 6 months after KTx, with no significant difference between Group D (healthy control group) and pre-surgery or Group C (2 years after KTx group). The pediatric donor group and the adult donor group reached immune homeostasis 3 months after the operation. Immune homeostasis is maintaining a balance between immune tolerance and immunogenicity. There was no significant difference in graft function between the pediatric and adult donor groups before surgery, 1 day after surgery, 1 week after surgery, 2 weeks after surgery, and 1 month after surgery; however, graft function was significantly better in the pediatric donor group compared with the adult donor group at 3 mouths (eGFR: 51.7 (40.4-66.2) vs 73.0 (55.7-90.2), P=0.008<0.05) and 6 months (eGFR: 52.2 (37.5-62.8) vs 80.5 (64.1-90.4), P<0.001) after surgery. Pediatric donor kidneys reached immune homeostasis 3 months after surgery, with better graft function at this time compared with adult donor kidneys. The proportion of Tregs/CD4⁺ T cells in recipients with a pulmonary infection after KTx was lower than in those with infection recovery. CONCLUSIONS Expanding the use of pediatric kidneys should be further explored by the transplantation community. The proportion of Tregs/CD4⁺ T cells in recipients with a pulmonary infection after KTx was lower than in those with infection recovery.

A Multiparametric Nomogram for Predicting Delayed Graft Function in Adult Recipients of Pediatric Donor Kidneys.

BACKGROUND: Delayed graft function (DGF) is one of the most common postoperative complications after kidney transplantation. The ability to predict DGF after transplantation can greatly aid clinical decision-making. Several models have been proposed to predict DGF in adult recipients of adult donor kidneys, but there is currently no model to predict DGF in adult recipients of pediatric donor transplants. Therefore, based on our medical records, we retrospectively investigated the pretransplant risk factors of DGF in transplants from pediatric donors to adult recipients. METHODS: Our center is in compliance with national laws, the Declaration of Istanbul, and the Helsinki Congress. The donors used by us were organs donated after the death of citizens and the participants were neither paid nor coerced. A retrospective review of 84 adult patients who received pediatric donor kidneys at a single center from April 4, 2015 to November 17, 2021 was conducted to investigate the pretransplant risk factors for the development of DGF. RESULTS: DGF was observed in 45 of 68 patients (66.17%) in the training group and 9 of 16 patients (56.25%) in the validation group. Multivariate logistic analysis showed that kidney donor profile index, cold ischemia time, number of human leukocyte antigen mismatches, and pretransplant dialysis duration were significant independent risk factors for DGF. By integrating these 4 factors, we constructed a nomogram model to predict DGF. According to the prediction model, the area under the curve of DGF of the training group and validation group was 0.899 and 0.905, respectively. CONCLUSION: We have constructed a novel, reliable, and accurate visual nomogram that provides a practical tool for predicting DGF in adult recipients of pediatric donor kidneys.

Mild hypothermia ameliorates hepatic ischemia reperfusion injury by inducing RBM3 expression.

Liver ischemia reperfusion injury (IRI) is a serious complication of certain liver surgeries, and it is difficult to prevent. As a potential drug-free treatment, mild hypothermia has been shown to promote positive outcomes in patients with IRI. However, the protective mechanism remains unclear. We established in vivo and in vitro models of hepatic ischemia reperfusion (IR) and mild hypothermia pretreatment. Hepatocytes were transfected with RNA-binding motif protein 3 (RBM3) overexpression plasmids, and IR was performed. Cell, culture medium, blood and tissue samples were collected to assess hepatic injury, oxidative stress, apoptosis and changes in RBM3 expression in the liver. Upregulation of RBM3 expression by mild hypothermia reduced the aminotransferase release, liver tissue injury and mitochondrial injury induced by liver IR. Hepatic IR-induced p38 and c-Jun N-terminal kinase (JNK) signaling pathway activation, oxidative stress injury and apoptosis could be greatly reversed by mild hypothermia. Overexpression of RBM3 mimicked the hepatoprotective effect of mild hypothermia. Mild hypothermia protects the liver from ischemia reperfusion-induced p38 and JNK signaling pathway activation, oxidative stress injury and apoptosis through the upregulation of RBM3 expression.

Prediction model of delayed graft function based on clinical characteristics combined with serum IL-2 levels.

BACKGROUND: Kidney transplantation is an effective treatment for end-stage renal disease (ESRD). Delayed graft function (DGF) is a common complication after kidney transplantation and exerts substantial effects on graft function and long-term graft survival. Therefore, the construction of an effective model to predict the occurrence of DGF is particularly important. METHODS: Seventy-one patients receiving their first kidney transplant at the First Affiliated Hospital of Nanchang University from October 2020 to October 2021 were enrolled in the discovery cohort. Based on clinical characteristics and serum markers, a logistic regression model was used to simulate the risk of DGF in the discovery cohort. The DGF prediction model was named the prediction system and was composed of risk factors related to DGF. Thirty-two patients receiving a kidney transplant at the First Affiliated Hospital of Nanchang University from October 2021 to February 2022 were enrolled in the validation cohort. The validation cohort was used to verify the accuracy and reliability of the prediction model. RESULTS: Cold ischemia time (CIT), donor history of diabetes mellitus, donor interleukin-2 (IL-2) level and donor terminal creatinine level constitute the prediction system. In the validation test, the area under the receiver operating characteristic curve (AUC) was 0.867 for the prediction system, and good calibration of the model was confirmed in the validation cohort. CONCLUSIONS: This study constructed a reliable and highly accurate prediction model that provides a practical tool for predicting DGF. Additionally, IL-2 participates in the kidney injury process and may be a potential marker of kidney injury.

Prognostic Analysis of Differentially Expressed DNA Damage Repair Genes in Bladder Cancer.

Bladder cancer (BCa) is the tenth most common tumor in humans. DNA damage repair genes (DDRGs) play important roles in many malignant tumors; thus, their functions in BCa should also be explored. We performed a comprehensive analysis of the expression profiles of DDRGs in 410 BCa tumors and 19 normal tissues from The Cancer Genome Atlas database. We identified 123 DDRGs differentially expressed between BCa tumors and normal tissues, including 95 upregulated and 28 downregulated genes. We detected 22 DDRGs associated with overall survival (OS) of patients with BCa by performing univariate Cox regression analysis. To explore the interactions between OS-associated DDRGs, we constructed a PPI network, which showed that the top six DDRGs (CDCA2, FOXM1, PBK, RRM2, ORC1, and HDAC4) with the highest scores in the PPI network might play significant roles in OS of BCa. Moreover, to investigate the latent regulatory mechanism of these OS-associated DDRGs, we analyzed the transcription factors (TFs)-DDRGs regulatory network. The core seven TFs (NCAPG, DNMT1, LMNB1, BRCA1, E2H2, CENPA, and E2F7) were shown to be critical regulators of the OS-related DDRGs. The 22 DDRGs were incorporated into a stepwise multivariable Cox analysis. Then, we built the index of risk score based on the expression of 8 DDRGs (CAD, HDAC10, JDP2, LDLR, PDGFRA, POLA2, SREBF1, and STAT1). The p-value < 0.0001 in the Kaplan-Meier survival plot and an area under the ROC curve (AUC) of 0.771 in TCGA-BLCA training dataset suggested the high specificity and sensitivity of the prognostic index. Furthermore, we validated the risk score in the internal TCGA-BLCA and an independent GSE32894 dataset, with AUC of 0.743 and 0.827, respectively. More importantly, the multivariate Cox regression and stratification analysis demonstrated that the predictor was independent of various clinical parameters, including age, tumor stage, grade, and number of positive tumor lymph nodes. In summary, a panel of 8 DNA damage repair genes associated with overall survival in bladder cancer may be a useful prognostic tool.

Curculigoside mitigates hepatic ischemia/reperfusion-induced oxidative stress, inflammation, and apoptosis via activation of the Nrf-2/HO-1 pathway.

Curculigoside has been shown to decrease oxidative stress and inflammatory reactions in many disorders, but its effects during hepatic ischemia-reperfusion injury (IRI) remain unknown. This research aims to determine the protective role and the potential mechanism of action of curculigoside in hepatic IRI. Here, a well-established rat model of partial warm IRI was constructed; serum ALT/AST and H&E staining were employed to assay the extent of liver injury; the superoxide dismutase, malondialdehyde, IL-6, and TNF-α contents were determined using the corresponding kits; the apoptosis index was evaluated by TUNEL staining; and the expression of Nrf-2, HO-1, and apoptosis-associated proteins was detected by qRT-PCR and Western blotting. The results showed that curculigoside pretreatment effectively mitigated hepatic IRI, as demonstrated by decreases in the levels of serum aminotransferases, hepatocellular necrosis and apoptosis, oxidative stress markers, infiltration of inflammatory cells, and secretion of proinflammatory cytokines. Mechanistically, the expression of Nrf-2 and HO-1 was greatly suppressed by hepatic IRI and reactivated by curculigoside. Furthermore, cotreatment with ML-385, an inhibitor of Nrf-2, counteracted the protective effect of curculigoside against hepatic IRI. The results of our study show that curculigoside plays a protective role in hepatic IRI by inhibiting oxidative stress, inflammation, and apoptosis and that its effects may be associated with activation of the Nrf-2/HO-1 pathway.

Machine learning-based method for tacrolimus dose predictions in Chinese kidney transplant perioperative patients.

WHAT IS KNOWN AND OBJECTIVES: Tacrolimus (TAC), a first-line immunosuppressant in solid-organ transplant, has a narrow therapeutic window and large inter-individual variability, which affects its use in clinical practice. Successful predictions using machine learning algorithms have been reported in several fields. However, a comparison of 10 machine learning model-based TAC pharmacogenetic and pharmacokinetic dosing algorithms for kidney transplant perioperative patients of Chinese descent has not been reported. The objective of this study was to screen and establish an appropriate machine learning method to predict the individualized dosages of TAC for perioperative kidney transplant patients. METHODS: The records of 2551 patients were collected from three transplant centres, 80% of which were randomly selected as a 'derivation cohort' to develop the dose prediction algorithm, while the remaining 20% constituted a 'validation cohort' to validate the final algorithm selected. Important features were screened according to our previously established population pharmacokinetic model of tacrolimus. The performances of the algorithms were evaluated and compared using R-squared and the mean percentage in the remaining 20% of patients. RESULTS AND DISCUSSION: This study identified several factors influencing TAC dosage, including CYP3A5 rs776746, CYP3A4 rs4646437, haematocrit, Wuzhi capsules, TAC daily dose, age, height, weight, post-operative time, nifedipine and the medication history of the patient. According to our results, among the 10 machine learning models, the extra trees regressor (ETR) algorithm showed the best performance in the training set (R-squared: 1, mean percentage within 20%: 100%) and test set (R-squared: 0.85, mean percentage within 20%: 92.77%) of the derivation cohort. The ETR model successfully predicted the ideal TAC dosage in 97.73% of patients, especially in the intermediate dosage range (>5 mg/day to <8 mg/day), whereby the ideal TAC dosage could be successfully predicted in 99% of the patients. WHAT IS NEW AND CONCLUSION: The results indicated that the ETR algorithm, which was chosen to establish the dose prediction model, performed better than the other nine machine learning models. This study is the first to establish ETR algorithms to predict TAC dosage. This study will further promote the individualized medication of TAC in kidney transplant patients in the future, which has great significance in ensuring the safety and effectiveness of drug use.

Alterations of thyroid microbiota across different thyroid microhabitats in patients with thyroid carcinoma.

BACKGROUND: In recent years, the incidence rate of Thyroid carcinoma (TC) has been increasing worldwide. Thus, research on factors of TC carcinogenesis may promote TC prevention and decrease the incidence rate. There are several studies targeting the correlation between gut microbiota and thyroid disease. Carcinogenesis of several malignancies is influenced by microbiota. However, thyroid microbiome of TC has not been revealed. This study investigated thyroid microbiota in different TC microhabitats. METHODS: We performed 16s rRNA gene sequencing using tumor tissues and matched peritumor tissues from 30 patients with TC to characterize thyroid microbiota. RESULTS: The richness and diversity of thyroid microbiota were lower in TC tumor samples than in matched peritumor tissues. At the genus level, the core microbiota of thyroid included Sphingomonas, Comamonas, Acinetobacter, Pseudomonas, Microvirgula, and Soonwooa. The abundance of Sphingomonas and Aeromonas was significantly increased in tumor tissues, while the abundance of Comamonas, Acinetobacter, and Peptostreptococcus was significantly enhanced in peritumor tissues. The combination of Comamonas and Sphingomonas could discriminate tumor samples from peritumor samples with an area under the curve (AUC) of 0.981 (95% confidence interval [CI] 0.949-1.000). The abundance of Sphingomonas was significantly higher in N1 stage than in N0 stage. Sphingomonas could distinguish between N0 and N1 stage with an AUC of 0.964 (95% CI 0.907-1.000). CONCLUSIONS: The microbial diversity and composition were significantly different between peritumor and tumor microhabitats from patients with TC, which may eventually affect TC carcinogenesis and progression. The combination of Comamonas and Sphingomonas could serve as a powerful biomarker for discrimination between tumor and peritumor tissues. Furthermore, the higher abundance of Sphingomonas was correlated with lymph node metastasis, indicating that the abundance of Sphingomonas may indicate a poor prognosis for TC patients, and Sphingomonas may play a role in promoting TC progression.

Intake of Dietary Fiber From Grains and the Risk of Hypertension in Late Midlife Women: Results From the SWAN Study.

Background: The possible effects of dietary fiber intake on hypertension have not been clarified fully. The association of dietary fiber intake with hypertension risk in midlife women was analyzed in this study. Methods: Baseline data were obtained from the Study of Women's Health Across the Nation (SWAN). Smooth curve, linear regression, and logistic regression analyses were performed to investigate the associations of four indices of daily dietary estimate (DDE) of dietary fiber (dietary fiber intake, dietary fiber intake from beans, dietary fiber intake from vegetables/fruit, and dietary fiber intake from grains) with blood pressure in midlife women. For this research purpose, diastolic blood pressure (DBP) ≥90 mmHg was defined as diastolic hypertension, and systolic blood pressure (SBP) ≥140 mmHg was defined as systolic hypertension. Results: This study included 2,519 participants with an average age of 46. The smooth curve showed approximate negative correlations between three fiber indices (DDE dietary fiber, DDE fiber from vegetables/fruit, and DDE fiber from grains) and blood pressure, including DBP and SBP (all P < 0.005). There were also approximate negative correlations between two fiber indices (DDE dietary fiber and DDE fiber from grains) and the risk of diastolic hypertension and systolic hypertension (all P < 0.05). Furthermore, multiple linear regression analysis suggested that DDE dietary fiber (Sß = -0.057, 95% CI -0.194 - -0.012, P = 0.027), DDE fiber from vegetables/fruit (Sß = -0.046, 95% CI -0.263 - -0.007, P = 0.039), and DDE fiber from grains (Sß = -0.073, 95% CI -0.600 - -0.099, P = 0.006, Model 4) were still negatively correlated with DBP after adjusting for confounding factors. Only DDE fiber from grains was independently and negatively associated with SBP (Sß = -0.060, 95% CI -0.846 - -0.093, P = 0.015) after these same confounding factors were adjusted for. Importantly, multiple logistic regression analysis suggested that only higher DDE fiber from grains was independently associated with a reduced risk of diastolic hypertension (OR = 0.848, 95% CI 0.770-0.934, P = 0.001, Model 4) and systolic hypertension (OR = 0.906, 95% CI 0.826-0.993, P = 0.034, Model 4) after the adjustments were made for confounding factors. Conclusions: We found that dietary fiber intake, especially DDE fiber from grains, contributes to a lower risk of systolic hypertension and diastolic hypertension in midlife women.

Comparison of Surgical Techniques in Living Donor Nephrectomy: A Systematic Review and Bayesian Network Meta-Analysis.

BACKGROUND The aim of this study was to compare and evaluate surgical techniques used for living donor nephrectomy (LDN). MATERIAL AND METHODS We performed a meta-analysis to compare 4 surgical techniques: open LDN (OLDN), laparoscopic LDN (LLDN), hand-assisted LLDN (HALLDN), and robot-assisted LLDN (RLDN). RESULTS No significant differences were found among these surgical techniques in terms of BMI, donor postoperative complications, 1-year graft survival, and DGF. Compared to the OLDN, the other 3 surgical techniques preferred to harvest the left kidney. When the right kidney was chosen as a donor, OLDN was the first-choice surgical technique. EBL was significantly lower in the HALLDN, LLDN, and RLDN groups when compared to the OLDN group. However, operative time and WIT were significantly shorter in the OLDN group. The RLDN group had an increased rate of donor intraoperative complications and a significantly lower VAS on day 1. The OLDN group required more morphine intake than the LLDN group. The length of hospital stay was significantly longer and AR was significantly higher in the OLDN group than in the LLDN and HALLDN groups. CONCLUSIONS There are no significant differences in donor postoperative complications, recipient DGF, and graft survival among the 4 surgical techniques. OLDN reduces WIT and operation time, but increases EBL and AR. RLDN and LLDN reduce the length of hospital stay, morphine intake, and VAS, and thus accelerate recovery. However, RLDN is associated with increased intraoperative complications.

TAK242 suppresses the TLR4 signaling pathway and ameliorates DCD liver IRI in rats.

Ischemia­reperfusion injury (IRI) is a notable cause of tissue damage during surgical procedures and a major risk factor in graft dysfunction in liver transplantation. Livers obtained from donors after circulatory death (DCD) are prone to IRI and toll­like receptor 4 (TLR4) serves a prominent role in the inflammatory response associated with DCD liver IRI. The present study was designed to investigate whether TAK242, a specific TLR4 inhibitor, improves hepatic IRI following a DCD graft and to investigate its underlying protective mechanisms. Male Sprague­Dawley rats were randomized into 4 groups: Control, TAK242, DCD and DCD+TAK242 groups. Rats were pretreated with TAK242 or its vehicle for 30 min, then the livers were harvested without warm ischemia (control group and TAK242 group) or with warm ischemia in situ for 30 min. The livers were stored in cold University of Wisconsin solution for 24 h and subsequently perfused for 60 min with an isolated perfused rat liver system. Rat liver injury was evaluated thereafter. When compared with the DCD group, DCD livers with TAK242 pretreatment displayed significantly improved hepatic tissue injury and less tissue necrosis (P<0.05). Compared with DCD livers, mechanistic experiments revealed that TAK242 pretreatment alleviated mitochondrial dysfunction, reduced reactive oxygen species and malondialdehyde levels and inhibited apoptosis. Additionally, TAK242 significantly inhibited the IRI­associated inflammatory response, indicated by the decreased expression of TLR4, interleukin (IL)­1ß, IL­6 and cyclooxygenase 2 at the mRNA and protein levels (P<0.05). TAK242 ameliorates DCD liver IRI via suppressing the TLR4 signaling pathway in rats. The results of the present study have revealed that TAK242 pretreatment harbors a potential benefit for liver transplantation.

Downregulation of Blimp1 inhibits the maturation of bone marrow-derived dendritic cells.

Modulation of differentiation of dendritic cells (DCs), which are derived from bone marrow cells, may influence their maturation and consequently regulate their ability to present antigens to alloreactive T lymphocytes. B lymphocyte­induced maturation protein­1 (Blimp1) is a master regulator of immunocyte differentiation, which has been investigated for its effect on DCs. In the present study, a lentivirus was used as a vector to transduce Blimp1­short hairpin (sh)RNA into primary bone marrow cells during their differentiation to DCs. Lentiviral­mediated Blimp1­shRNA (lenti­shRNA­Blimp1) had a transduction efficiency of >60% in DC precursors. Lenti­shRNA­Blimp1 significantly downregulated the expression levels of Blimp1 and modulated the expression of its target proteins, including class II major histocompatibility complex (MHC) transactivator, c­myc and interleukin­6. Although lenti­shRNA­Blimp1 did not interfere with the differentiation of bone marrow cells to DCs, it inhibited DC maturation by decreasing the expression of surface MHC­II molecules, but not the expression of MHC­I molecules and co­stimulatory molecules [cluster of differentiation (CD)80/CD86]. Subsequently, alloreactive T cell proliferation was alleviated and regulatory T cells were expanded in response to lenti­shRNA­Blimp1. A toxicity assay indicated that the morphology and proliferation of cultured DCs were mildly influenced by the lentiviral vector, indicating that the use of alternative vectors with minimal or no toxicity could be investigated in future studies. In conclusion, transduction with lenti­shRNA­Blimp1 modulated the maturation of DCs via MHC­II molecule suppression and inhibited alloreactive T cell activation. The present findings supported the application of Blimp1­based intervention as a novel approach to induce immature DCs for further immunological research.

Mild hypothermia pretreatment protects against liver ischemia reperfusion injury via the PI3K/AKT/FOXO3a pathway.

Mild hypothermia is known to protect against ischemia and reperfusion (IR) injury. The exact mechanisms of the protection are not fully understood. Forkhead box O3 (FOXO3a) has been defined as a critical mediator in cellular processes, including oxidative stress, apoptosis, inflammation, cell death and DNA repair; however, the protection function in mild hypothermia has not been reported previously. The current study was designed to investigate the function of phosphoinositide 3­kinase (PI3K)/protein kinase B (AKT)/FOXO3a pathway in pretreatment with mild hypothermia during IR injury. Additionally, PI3K/AKT/FOXO3a signaling was inhibited using Ly294002 and the effect on the protective function of mild hypothermia pretreatment was evaluated. Furthermore, the apoptotic and inflammatory response induced by the IR injury was evaluated. Liver IR injury induced a significant increase in the level of apoptosis and inflammatory responses. However, pretreatment with mild hypothermia increased phospho (p)­AKT and p­FOXO3a following IR injury, and significantly reduced apoptosis and inflammatory cytokines release. However, inhibiting p­AKT and p­FOXO3a using Ly294002 suppressed the liver protection produced by mild hypothermia. In conclusion, these findings indicated that mild hypothermia pretreatment exhibited liver protective effects against IR injury associated with suppressing inflammatory cytokine release and apoptosis via the PI3K/AKT/FOXO3a pathway.

[The protective effects on allografts of adeno-associated heme-oxygenase-1 gene therapy against chronic rejection injury].

OBJECTIVE: To investigate the protective effects on allografts and the possible mechanism of adeno-associated heme-oxygenase-1 (AdHO-1) gene therapy against chronic rejection injury. METHODS: Ex vivo AdHO-1 gene therapy was performed in vascular and renal transplantation models. The structure and function, the expression of therapeutic genes and proteins, and the immune modulation were analyzed. RESULTS: AdHO-1 gene therapy protected renal transplant against chronic rejection, but the effect was not as remarkable as that in vascular transplant. The transfected empty vehicle aggravated chronic rejection damage in renal transplantation. AdHO-1 decreased the infiltration of macrophages and CD4(+) T cells. CONCLUSIONS: AdHO-1 gene therapy can lessen damage of chronic rejection in allografts. It plays roles by protecting transplants, down-regulating immune response and inducing immune deviation.

Effects of ischemic preconditioning on cyclinD1 expression during early ischemic reperfusion in rats.

AIM: To observe the effect of ischemic preconditioning on cyclinD1 expression in rat liver cells during early ischemic reperfusion. METHODS: Fifty-four SD rats were randomly divided into ischemic preconditioning group (IP), ischemia/reperfusion group (IR) and sham operation group (SO). The IP and IR groups were further divided into four sub-groups (n = 6). Sham operation group (SO) served as the control group (n = 6). A model of partial liver ischemia/reperfusion was used, in which rats were subjected to liver ischemia for 60 min prior to reperfusion. The animals in the IP group underwent ischemic preconditioning twice for 5 min each time prior to the ischemia/reperfusion challenge. After 0, 1, 2, and 4 h of reperfusion, serum and liver tissue in each group were collected to detect the level of serum ALT, liver histopathology and expression of cyclinD1 mRNA and protein. Flow cytometry was used to detect cell cycle as the quantity indicator of cell regeneration. RESULTS: Compared with IR group, IP group showed a significantly lower ALT level in 1 h to 4 h sub-groups (P < 0.05). Proliferation index (PI) indicated by the S-phase and G2/M-phase ratio [(S+G2/M)/(G0/G1+S+G2/M)] was significantly increased in IP group at 0 and 1 h (26.44 +/- 7.60% vs 18.56 +/- 6.40%, 41.87 +/- 7.27% vs 20.25 +/- 6.70%, P < 0.05). Meanwhile, cyclinD1 protein expression could be detected in IP group. But in IR group, cyclinD1 protein expression occurred 2 h after reperfusion. The expression of cyclinD1 mRNA increased significantly in IP group at 0 and 1 h (0.568 +/- 0.112 vs 0.274 +/- 0.069, 0.762 +/- 0.164 vs 0.348 +/- 0.093, P < 0.05). CONCLUSION: Ischemic preconditioning can protect liver cells against ischemia/reperfusion injury, which may be related to cell proliferation and expression of cyclinD1 during early ischemic reperfusion.