Metastasis and basement membrane-related signature enhances hepatocellular carcinoma prognosis and diagnosis by integrating single-cell RNA sequencing analysis and immune microenvironment assessment.

BACKGROUND: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and second leading cause of cancer-related deaths worldwide. The heightened mortality associated with HCC is largely attributed to its propensity for metastasis, which cannot be achieved without remodeling or loss of the basement membrane (BM). Despite advancements in targeted therapies and immunotherapies, resistance and limited efficacy in late-stage HCC underscore the urgent need for better therapeutic options and early diagnostic biomarkers. Our study aimed to address these gaps by investigating and evaluating potential biomarkers to improve survival outcomes and treatment efficacy in patients with HCC. METHOD: In this study, we collected the transcriptome sequencing, clinical, and mutation data of 424 patients with HCC from The Cancer Genome Atlas (TCGA) and 240 from the International Cancer Genome Consortium (ICGC) databases. We then constructed and validated a prognostic model based on metastasis and basement membrane-related genes (MBRGs) using univariate and multivariate Cox regression analyses. Five immune-related algorithms (CIBERSORT, QUANTISEQ, MCP counter, ssGSEA, and TIMER) were then utilized to examine the immune landscape and activity across high- and low-risk groups. We also analyzed Tumor Mutation Burden (TMB) values, Tumor Immune Dysfunction and Exclusion (TIDE) scores, mutation frequency, and immune checkpoint gene expression to evaluate immune treatment sensitivity. We analyzed integrin subunit alpha 3 (ITGA3) expression in HCC by performing single-cell RNA sequencing (scRNA-seq) analysis using the TISCH 2.0 database. Lastly, wound healing and transwell assays were conducted to elucidate the role of ITGA3 in tumor metastasis. RESULTS: Patients with HCC were categorized into high- and low-risk groups based on the median values, with higher risk scores indicating worse overall survival. Five immune-related algorithms revealed that the abundance of immune cells, particularly T cells, was greater in the high-risk group than in the low-risk group. The high-risk group also exhibited a higher TMB value, mutation frequency, and immune checkpoint gene expression and a lower tumor TIDE score, suggesting the potential for better immunotherapy outcomes. Additionally, scRNA-seq analysis revealed higher ITGA3 expression in tumor cells compared with normal hepatocytes. Wound healing scratch and transwell cell migration assays revealed that overexpression of the MBRG ITGA3 enhanced migration of HCC HepG2 cells. CONCLUSION: This study established a direct molecular correlation between metastasis and BM, encompassing clinical features, tumor microenvironment, and immune response, thereby offering valuable insights for predicting clinical outcomes and immunotherapy responses in HCC.

Lysine demethylase KDM3A alleviates hyperoxia-induced bronchopulmonary dysplasia in mice by promoting ETS1 expression.

Bronchopulmonary dysplasia (BPD) is the most common chronic lung disease among neonates, with increasing morbidity and mortality. This study aims to investigate the effect and mechanism of lysine demethylase 3A (KDM3A) on hyperoxia-induced BPD. Hyperoxia-induced BPD mouse and alveolar epithelial cell models were constructed. The effects of hyperoxia on lung development were evaluated by histological and morphological analysis. The levels of KDM3A, E26 transformation specific-1 (ETS1), H3 lysine 9 dimethylation (H3K9me2), and endoplasmic reticulum (ER) stress-related indexes were quantified by RT-qPCR, Western blot, and IF staining. Cell apoptosis was assessed by flow cytometry and TUNEL staining. Transfection of oe-ETS1, oe-KDM3A, and sh-ETS1 was applied in hyperoxia-induced alveolar epithelial cells to explore the mechanism of the KDM3A/ETS1 axis in hyperoxia-induced apoptosis. KDM3A inhibitor IOX1 was applied to validate the in vivo effect of KDM3A in hyperoxia-induced BPD mice. The results displayed that hyperoxia-induced BPD mice showed reduced body weight, severe destruction of alveolar structure, decreased radial alveolar count (RAC), and increased mean linear intercept (MLI) and mean alveolar diameter (MAD). Further, hyperoxia induction down-regulated ETS1 expression, raised ER stress levels, and increased apoptosis rate in BPD mice and alveolar epithelial cells. However, transfection of oe-ETS1 improved the above changes in hyperoxia-induced alveolar epithelial cells. Moreover, transfection of oe-KDM3A up-regulated ETS1 expression, down-regulated H3K9me2 expression, inhibited ER stress, and reduced apoptosis rate in hyperoxia-induced alveolar epithelial cells. In addition, transfection of sh-ETS1 reversed the inhibitory effect of KDM3A on hyperoxia-induced apoptosis by regulating ER stress. In vivo experiments, KDM3A inhibitor IOX1 intervention further aggravated BPD in newborn mice. In a word, KDM3A alleviated hyperoxia-induced BPD in mice by promoting ETS1 expression.

Identification and validation of immune-related methylated genes as diagnostic and prognostic biomarkers of nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a common malignancy occurring in the head and neck. Identification of immune-related methylated biomarkers might be helpful for NPC detection and prognostic evaluation. METHODS: A co-methylation network based on WGCNA was constructed to identify modules associated with NPC and immune cells. In combination with differentially expressed genes (DEGs) and immune-related genes from ImmPort database, the candidate immune-related methylated genes (IRMGs) were obtained. RESULTS: Our combined analysis identified 12 IRMGs. Among them, both the methylation and mRNA expression of CCL28, CSK, and PRKCB were correlated with the infiltration of B cells. CD1D, CR2, and GDF10 were favorable markers. Demethylation experiments validated that downregulation of GDF10, PRKCB, SLC40A1, and TGFBR3 in NPC resulted from promoter hypermethylation. Additionally, a diagnostic model was developed and exhibited high discriminative accuracy. CONCLUSIONS: These results provided a group of immune-related methylated biomarkers that may help with the diagnosis and prognosis of NPC.

ETS1 Ameliorates Hyperoxia-Induced Bronchopulmonary Dysplasia in Mice by Activating Nrf2/HO-1 Mediated Ferroptosis.

PURPOSE: Bronchopulmonary dysplasia (BPD) is associated with hyperoxia-induced oxidative stress-associated ferroptosis. This study examined the effect of E26 oncogene homolog 1 (ETS1) on oxidative stress-associated ferroptosis in BPD. METHODS: Hyperoxia-induced A549 cells and neonatal mice were used to establish BPD models. The effects of ETS1 on hyperoxia-induced ferroptosis-like changes in A549 cells were investigated by overexpression of ETS1 plasmid transfection and erastin treatment. Glucose consumption, lactate production, and NADPH levels were assessed by the glucose, lactate, and NADP+/NADPH assay kits, respectively. The potential regulatory relationship between ETS1 and Nrf2/HO-1 was examined by treating hyperoxia-induced A549 cells with the Nrf2 inhibitor ML385. ETS1 effect on the Nrf2 promoter was explored by dual-luciferase reporter and chromatin immunoprecipitation assay. The effect of ETS1 on the symptoms of BPD mice was examined by injecting an adenovirus overexpressing ETS1. RESULTS: ETS1 overexpression increased hyperoxia-induced cell viability, glucose consumption, lactate production, and NADPH levels and reduced inflammation and apoptosis in A549 cells. In animal experiments, ETS1 overexpression prevented weight loss, airway enlargement, and reductions in radial alveolar counts in BPD mice, while reducing the mean linear intercept, mean alveolar diameter and inflammation. ETS1 overexpression suppressed PTGS2 and CHAC1 expression, reduced ROS, MDA and ferrous iron (Fe2+) production and increased GSH levels in hyperoxia-induced A549 cells and BPD mice. In addition, ETS1 can bind to the Nrf2 promoter region and thus promote Nrf2 transcription. ETS1 overexpression increased the mRNA and protein levels of Nrf2, HO-1, xCT, and GPX4 in hyperoxia-induced A549 cells and BPD mice. In hyperoxia-induced A549 cells, erastin and ML385 treatment abolished the effect of ETS1 overexpression. CONCLUSION: ETS1 is important in oxidative stress-related ferroptosis in a hyperoxia-induced BPD model, and the effect is partially mediated by the Nrf2/HO-1 axis.

Prognostic, Immunological, and Mutational Analysis of MTA2 in Pan-Cancer and Drug Screening for Hepatocellular Carcinoma.

BACKGROUND: Metastasis-associated protein 2 (MTA2) is a member of the metastasis-associated transcriptional regulator family and is a core component of the nucleosome remodeling and histone deacetylation complex. Despite growing evidence that MTA2 plays a crucial role in the tumorigenesis of certain cancers, no systematic pan-cancer analysis of MTA2 is available to date. Therefore, the aim of our study is to explore the prognostic value of MTA2 in 33 cancer types and to investigate its potential immune function. METHODS: by comprehensive use of databases from TCGA, GTEx, GEO, UCSC xena, cBioPortal, comPPI, GeneMANIA, TCIA, MSigDB, and PDB, we applied various bioinformatics approaches to investigate the potential role of MTA2, including analyzing the association of MTA2 with MSI, prognosis, gene mutation, and immune cell infiltration in different tumors. We constructed a nomogram in TCGA-LIHC, performed single-cell sequencing (scRNA-seq) analysis of MTA2 in hepatocellular carcinoma (HCC), and screened drugs for the treatment of HCC. Finally, immunohistochemical experiments were performed to verify the expression and prognostic value of MTA2 in HCC. In vitro experiments were employed to observe the growth inhibition effects of MK-886 on the HCC cell line HepG2. RESULTS: The results suggested that MTA2 was highly expressed in most cancers, and MTA2 expression was associated with the prognosis of different cancers. In addition, MTA2 expression was associated with Tumor Mutation Burden (TMB) in 12 cancer types and MSI in 8 cancer types. Immunoassays indicated that MTA2 positively correlated with activated memory CD4 T cells and M0 macrophage infiltration levels in HCC. ScRNA-seq analysis based on the GEO dataset discovered that MTA2 was significantly expressed in T cells in HCC. Finally, the eXtreme Sum (Xsum) algorithm was used to screen the antitumor drug MK-886, and the molecular docking technique was utilized to reveal the binding capacity between MK-886 and the MTA2 protein. The results demonstrated excellent binding sites between them, which bind to each other through Π-alkyl and alkyl interaction forces. An immunohistochemistry experiment showed that MTA2 protein was highly expressed in HCC, and high MTA2 expression was associated with poor survival in HCC patients. MK-886 significantly inhibited the proliferation and induced cell death of HepG2 cells in a dose-dependent manner. CONCLUSIONS: Our study demonstrated that MTA2 plays crucial roles in tumor progression and tumor immunity, and it could be used as a prognostic marker for various malignancies. MK-886 might be a powerful drug for HCC.

Ferroptosis-Related lncRNAs Act as Novel Prognostic Biomarkers in the Gastric Adenocarcinoma Microenvironment, Immunotherapy, and Chemotherapy.

Ferroptosis, a form of programmed cell death akin to necrosis, is managed by iron and is distinguished by lipid peroxidation. Gastric cancer is a highly aggressive form of cancer, responsible for the third highest number of cancer-related deaths globally. Despite this, the potential of ferroptosis to predict the occurrence of this cancer is yet to be determined. In this research, a comprehensive examination was conducted to explore the link between long noncoding RNAs (lncRNAs) and ferroptosis, in order to uncover an lncRNA signature that can predict drug susceptibility and tumor mutational burden (TMB) in gastric adenocarcinoma. We conducted an in-depth analysis of the GC immune microenvironment and immunotherapy, with a particular focus on ferroptosis-related lncRNA prognostic biomarkers, and further explored the correlation between these factors and prognosis, immune infiltration, single nucleotide variation (SNV), and drug sensitivity for gastric adenocarcinoma patients. Through our investigations, we have discovered five lncRNA signatures related to ferroptosis that can accurately forecast the prognosis of gastric adenocarcinoma patients and also regulate the proliferation, migration, and occurrence of ferroptosis in gastric adenocarcinoma cells. In conclusion, this lncRNA signature associated with ferroptosis may be employed as a prognostic indicator for gastric adenocarcinoma, thus presenting a potential solution.

Safety and efficacy of using portable coagulation monitor for INR examination after left-sided mechanical prosthetic valve replacement.

BACKGROUND: Time in therapeutic range (TTR) is an index to assess the effectiveness of anticoagulation and is important to predict the risk of bleeding and thrombosis in patients taking warfarin. In recent years, the portable coagulation monitor, a point-of-care testing device for patients to perform self-management international normalized ratio (INR) examination, has provided an opportunity to improve the quality of oral warfarin treatment. In this study, we applied TTR to evaluate the safety and efficacy of the portable coagulation monitor for patients with oral anticoagulant warfarin after left-sided mechanical prosthetic valve (MPV) replacement. METHODS: It is a single-centre cohort study. From September 2019 to June 2021, a total of 243 patients who returned to our institution for outpatient clinic revisit at 3 months after left-sided MPV replacement, met the inclusion criteria and agreed to be followed up were included. Self-management group used portable coagulation monitor for INR examination, and patients in the conventional group had their INR monitored in routine outpatient visits. Clinical data of the patients would be recorded for the next 12 months, and results were compared between the two groups to assess the effect of the coagulation monitor on TTR and complications related to bleeding and thrombosis in patients with left-sided MPV replacement. RESULTS: A total of 212 individuals provided complete and validated INR data spanning of 1 year. Those who applied the portable coagulation monitor had higher TTR values and larger number of tests for INR. No significant differences were seen between the two groups in postoperative bleeding and thromboembolic complications, but portable coagulation monitor showed a trend toward fewer bleeding events. CONCLUSION: Portable devices for coagulation monitoring are safe and can achieve a higher TTR. Patients who use the portable coagulation monitor for home INR self-management can achieve a safe and effective warfarin therapy.

Lef1 is transcriptionally activated by Klf4 and suppresses hyperoxia-induced alveolar epithelial cell injury.

PURPOSE: Bronchopulmonary dysplasia (BPD) is a long-term respiratory condition. More than a quarter of extremely premature newborns are harmed by BPD. At present, there are no apparent effective drugs or treatments for the condition. In this study, we aimed to investigate the functional role and mechanism of lymphoid enhancer-binding factor 1 (Lef1) in BPD in vitro. MATERIALS AND METHODS: Blood samples from BPD patients and healthy volunteers were gathered, and an in vitro model of BPD was developed in alveolar epithelial cells (AECs) MLE-12 induced by hyperoxia. Then expression of krüppel-like factor 4 (KLF4/Klf4) and LEF1/Lef1 were evaluated. After Lef1 overexpressing plasmid and the vector were transfected into hyperoxia-induced MLE-12 cells, cell proliferation assays were carried out. Cell apoptosis was investigated by a flow cytometry assay, and apoptosis related proteins Bcl-2, cleaved-caspase 3 and 9 were analyzed by a western blot assay. The binding between Klf4 and Lef1 promoter predicted on the JASPAR website was verified using luciferase and ChIP assays. For further study of the mechanism of Klf4 and Lef1 in BPD, gain-of-function experiments were performed. RESULTS: The mRNA levels of KLF4/Klf4 and LEF1/Lef1 were diminished in clinical BPD serum samples and hyperoxia-induced MLE-12 cells. Overexpression of Lef1 stimulated AEC proliferation and suppressed AEC apoptosis induced by hyperoxia. Mechanically, Klf4 bound to Lef1's promoter region and aids transcription. Moreover, the results of gain-of-function experiments supported that Klf4 could impede AEC damage induced by hyperoxia via stimulating Lef1. CONCLUSION: Klf4 and Lef1 expression levels were declined in hyperoxia-induced AECs, and Lef1 could be transcriptionally activated by Klf4 and protect against hyperoxia-induced AEC injury in BPD. As a result, Lef1 might become a prospective therapeutic target for BPD.

Open Seldinger-guided peripheral femoro-femoral cannulation technique for totally endoscopic cardiac surgery.

BACKGROUND: The cannulation technique used in totally endoscopic cardiac surgery has a significant impact on the overall prognosis of patients. However, there are no large cohort studies to discuss it. Here we report on our research of using open Seldinger-guided technique to establish femoro-femoral cardiopulmonary bypass during totally endoscopic cardiac surgery and evaluate its safety and efficacy. METHODS: The institutional database from 2017 to 2020 was retrospectively reviewed to find cases in which totally endoscopic cardiac surgery was performed. We identified 214 consecutive patients who underwent totally endoscopic cardiac surgery with peripheral femoro-femoral cannulation. All patients underwent femoral artery cannulation. Of these, 201 were cannulated in the femoral vein and 13 were cannulated in the femoral vein combined with internal jugular cannulation. The technique involves surgically exposing the femoral vessel, setting up purse-string over the vessels and then inserting a guidewire into the femoral vessel without a vascular incision, followed by exchange of the guidewire with a cannula. RESULTS: Surgery indications included mitral valve disease in 82.71% (177/214), atrial septal defect in 11.68% (25/214) and tricuspid regurgitation in the remaining 5.61% (12/214). Hospital survival was 98.60% (211/214). There were no cases of stroke and postoperative limb ischaemia. No femoral vessel injuries or wound infections was observed. No late pseudoaneurysms were evident. CONCLUSION: The open Seldinger-guided femoro-femoral cannulation technique is effective and safe. We highly recommend this technique, given its safety, simplicity and speed under direct vision. The limited manipulation of the vessels under direct vision minimizes the risk of local complications.

Transcatheter and intraoperative device closure and surgical repair for atrial septal defect.

BACKGROUND: Transcatheter and intraoperative device closure for atrial septal defect (ASD) are widely applied to reduce the incision size and the potential for injury during cardiopulmonary bypass (CPB) in conventional surgical repair. No studies had been conducted to compare the safety and efficiency of these three treatments. METHODS: From January 2018 to April 2018, 87 patients with an isolated ASD who had undergone transcatheter device closure (n = 45), intraoperative device closure (n = 22) and surgical repair (n = 20) were retrospectively reviewed and further analyzed to compare these three treatments. RESULTS: The successful closure rate was similar in the three groups. There was a significant difference in aortic cross-clamping time, CPB duration and operative time between the surgical group and the device groups. The length of intensive care unit stay, postoperative mechanical ventilation time and length of hospital stay were shorter in the two device groups than in the surgical group. The incision was the most extended in the surgical group. Regarding major adverse events, no significant differences were found among the three groups. CONCLUSIONS: Transcatheter and intraoperative device closure and surgical repair for ASD are all safe and effective. Considering their respective disadvantages and advantages, the transcatheter approach may be the first choice for an isolated secundum ASD, the intraoperative approach may be the second choice, and surgical repair may be the last resort.

A Comparative Study of Perventricular and Percutaneous Device Closure Treatments for Isolated Ventricular Septal Defect: A Chinese Single-Institution Experience.

OBJECTIVES: To compare the advantages and disadvantages of perventricular and percutaneous procedures for treating isolated ventricular septal defect (VSD). METHODS: A total of 572 patients with isolated VSD were selected in our hospital between January 2015 and December 2016. The patients' median age and weight were five years (1-26 years) and 29 kg (9-55 kg), respectively. The median diameter of VSD was 6.0 mm (5-10 mm). Patients were divided into two groups. In group A, perventricular device closure was performed in 427 patients; in group B, 145 patients underwent percutaneous device closure. RESULTS: Four hundred twelve patients in group A and 135 patients in group B underwent successful closure. The total occlusion rate was 98.5% (immediately) and 99.5% (3-month follow-up) in group A, which were not significantly different from those in group B (97.7% and 100%, respectively). Patients in group A had longer intensive care unit (ICU) stay than those in group B, but patients in group B experienced significantly longer operative times than those in group A. The follow-up period ranged from 8 months to 1.5 year (median, 1 year). During the follow-up period, late-onset complete atrioventricular block occurred in two patients. No other serious complications were noted in the remaining patients. CONCLUSION: Both procedures are safe and effective treatments for isolated VSD. The percutaneous procedure has obvious advantages of shorter ICU stay and less trauma than the perventricular procedure. However, the perventricular procedure is simpler to execute, results in a shorter operative time, and avoids X-ray exposure.

A comparative study of perventricular and percutaneous device closure treatments for isolated ventricular septal defect: a chinese single-institution experience

Abstract Objectives: To compare the advantages and disadvantages of perventricular and percutaneous procedures for treating isolated ventricular septal defect (VSD). Methods: A total of 572 patients with isolated VSD were selected in our hospital between January 2015 and December 2016. The patients' median age and weight were five years (1-26 years) and 29 kg (9-55 kg), respectively. The median diameter of VSD was 6.0 mm (5-10 mm). Patients were divided into two groups. In group A, perventricular device closure was performed in 427 patients; in group B, 145 patients underwent percutaneous device closure. Results: Four hundred twelve patients in group A and 135 patients in group B underwent successful closure. The total occlusion rate was 98.5% (immediately) and 99.5% (3-month follow-up) in group A, which were not significantly different from those in group B (97.7% and 100%, respectively). Patients in group A had longer intensive care unit (ICU) stay than those in group B, but patients in group B experienced significantly longer operative times than those in group A. The follow-up period ranged from 8 months to 1.5 year (median, 1 year). During the follow-up period, late-onset complete atrioventricular block occurred in two patients. No other serious complications were noted in the remaining patients. Conclusion: Both procedures are safe and effective treatments for isolated VSD. The percutaneous procedure has obvious advantages of shorter ICU stay and less trauma than the perventricular procedure. However, the perventricular procedure is simpler to execute, results in a shorter operative time, and avoids X-ray exposure.

[Vascular bypass grafting combined with endovascular aortic repair for treatment of aortic dilatation disease].

OBJECTIVE: To summarize the clinical experience of vascular bypass grafting combined with endovascular aortic repair (EVAR) for aortic dilatation disease. METHODS: Between January 2008 and August 2011, 12 patients with aortic dilatation disease were treated with vascular bypass grafting combined with EVAR. Of 12 patients, 11 were male and 1 was female, aged 47-81 years (mean, 65.9 years). All cases were diagnosed through computed tomography angiography (CTA), including 1 case of Stanford type A dissection, 5 cases of Stanford type B dissection, 4 cases of aortic arch aneurysm, and 2 cases of abdominal aortic aneurysm. Eight patients received neck artery bypass grafting before EVAR, and 4 patients underwent femoral artery bypass grafting after EVAR. RESULTS: After operation, pulmonary infection occurred in 3 patients, renal insufficiency in 2 patients, cerebral infarction in 1 case, decreased hemoglobin and platelets in 7 cases, and poor healing of groin wound in 1 case. Eleven patients were followed up 3-42 months, with an average of 18.6 months. In 1 case undergoing EVAR of the thoracic and abdominal aorta, EVAR was performed again because new aneurysms formed at 6 months after operation, and the patient achieved good recovery after 3 months. CTA showed reduced false lumen, thrombosis formation, no endoleak, no deformation or displacement of stent, and anastomotic patency of artificial blood vessels in the other patients at 3, 6, and 12 months after operation. CONCLUSION: Vascular bypass grafting combined with EVAR can expand the indications for endovascular repair. It not only provides sufficient anchoring area, but also ensures the blood supply to vital organs, simplifies the surgical procedure, and reduces the difficulty of endovascular treatment.

[Outcome post orthotopic heart transplantation for patients with end-stage hypertrophic cardiomyopathy].

OBJECTIVE: To investigate the outcome of orthotopic heart transplantation for patient with end-stage hypertrophic cardiomyopathy. METHODS: This retrospective review analyzed the clinical data of nine patients (7 males) undergoing orthotopic heart transplantation for end-stage hypertrophic cardiomyopathy in our center. All patients received induced therapy protocols peri-operative and standard triple maintenance immunosuppressive therapy postoperative. RESULTS: One recipients developed acute renal failure due to renal artery embolism and allograft rejection in the early posttransplantive course, symptoms and signs were improved under continuous renal replacement therapy and steroid-pulse therapy, this patient died of sudden cardiac arrest at 32 months post transplantation. Another recipient developed demyelinating disease in frontal and parietal lobe and finally recovered with medical therapy. Eight patients survived the operation with good quality of life and there was no episode of rejection or infection or chronic graft arteriosclerosis during follow-up time. Three recipients developed left ventricular hypertrophy and there were no signs of grapg-vessel diseases in the survivals. CONCLUSION: Heart transplantation is the best therapeutic option for selected patients with end-stage hypertrophic cardiomyopathy.

[Mid- and long-term acute cardiac allograft rejection: clinical observation of 14 patients].

OBJECTIVE: To analyze the clinical features of mid- and long-term acute cardiac allograft rejection to improve the long-term clinical outcomes of the patients. METHODS: Fourteen recipients (11 males and 3 females) underwent orthotopic heart transplantation with standard immunosuppressive therapy protocols (3 cases) or induction therapy protocols (11 cases). Cyclosporine, azathioprine or mycophenolate mofetil, and prednisolone were applied as the maintenance immunosuppressive regimen. Acute graft rejection episodes occurred within 3 to 6 months in 1 case, within 6 months to 1 year in 3 cases, within 1 to 2 years in 3 cases, within 2 to 5 years in 6 cases, and above 5 years in 1 case. RESULTS: No significant difference was found in the incidence of late heart rejection between the patients receiving the two immunosuppressive therapy protocols. Immunosuppressants were withdrawn or spared in 8 recipients due to different causes. Nine recipients with steroid-sensitive acute cardiac allograft rejection were treated with steroid-pulse therapy, while the other 5 were treated with a short course of polyclonal antithymocyte antibodies because of steroid-resistant acute rejection; in 11 cases, azathioprine was converted to mycophenolate mofetil. Four of the 5 late deaths occurred in the recipients with steroid-resistant rejection. The surviving recipients had a good quality of life, and no recurrent episodes of rejection or infection were observed in the follow-up period. CONCLUSIONS: Late acute cardiac allograft rejection is associated mainly with patient compliance but not with early immunosuppressive therapy protocols. The episodes are rather severe and should be timely treated with steroid pulses or polyclonal antithymocyte antibodies.

[Individual immunosuppressive regime in heart transplantation with high risk].

OBJECTIVE: To Summarize the clinical experience of individual immunosuppressive regime in heart transplantation with high risk. METHODS: From September 2001 to December 2006, 51 cases with the complication of Hepatitis B viruses (HBV) infection, diabetes mellitus, renal dysfunction or pulmonary infection in perioperative period were analyzed retrospectively. All cases received daclizumab (Zenapax) induction therapy, and baseline triple immunosuppressive regime was consist of cyclosporine (CsA), azathioprine (Aza) or mycophenolate mofetil (MMF) and prednisone (Pred). Ten cases received HBV infection in preoperative period, the immunosuppressive protocol was emphasized on the use of MMF and the withdraw of Pred one month later in postoperation. Nine cases received diabetes mellitus in pre-operation, 4 cases had post-transplant diabetes mellitus. The immunosuppressive protocol was emphasized on the use of CsA rather than FK506, the use of Pred was less dosage, and the therapy of insulin was necessary. Sixteen cases had renal dysfunction in pre-operation, the use of MMF was routine but the use of CsA was delayed to the time 5 to 19 d postoperative. Twelve cases received pulmonary infection after allograft transplantation. The immunosuppressive agent was to be taped or suspended in therapy time. RESULTS: The liver function of the 10 cases with HBV infection was stable in 1 year follow-up, and 1 case received acute rejection after 13 months allograft transplantation. In the 6 months follow-up, the blood glucose level of the 13 cases with diabetes mellitus was stable, none of the cases suffered from acute rejection. In the one month follow-up, none of the 16 cases with renal dysfunction suffered from acute rejection, and the renal function was normal. Two of the 12 cases with the pulmonary infection were died of serious infection, others were survival. One case received acute rejection on the 17th day in postoperation. CONCLUSIONS: Low mortality can be realized by selecting appropriately individual immunosuppressive regime and the episode of acute rejection is rare.

[Effect of berberin hydrochloride on blood concentration of cyclosporine A in cardiac transplanted recipients].

OBJECTIVE: To observe the clinic effect of combined use of berberin hydrochloride (Ber) with cyclosporine A (CsA) on the blood concentration of CsA in heart transplanted recipients. METHODS: The blood concentration of CsA, liver-renal function and blood lipids in 22 heart transplanted recipients, who received Ber-CsA combined therapy, were measured. RESULTS: The whole blood steady state concentration of CsA, C0 and C2, in recipients after being treated with Ber-CsA significantly increased than those before applying Ber-CsA (P < 0.01), with the mean increment of 26% and 18% respectively; the dosage of CsA used decreased in 21 patients by 25-100 mg/d; and the Ber-CsA showed no significant effect on liver-renal function or blood lipids (P > 0.05). CONCLUSION: Combined use of CsA with Ber could markedly increase the blood concentration of CsA in heart transplanted recipients and reduce the dosage of CsA required, save the fee for medical service, and shows no obvious adverse reaction.

[A modified model of cervical heterotopic cardiac transplantation for chronic rejection research].

OBJECTIVE: To establish the modified model of cervical heterotopic cardiac transplantation in rats for investigation of cardiac chronic rejection. METHODS: Forty healthy male Wistar rats, aged 10 weeks, weighing 250-300 g, were applied as the donor group, and forty healthy male SD rats, aged 10 weeks, weighing 300-350 g, served as the recipient group. The donors' pulmonary artery was anastomosed to the recipients' right external jugular vein by non-suture cuff technique while the donors' innominate artery was anastomosed to the recipients' right common carotid artery by suture microvascular anastomosis. All recipients received cyclosporin to prevent acute allograft rejection. RESULTS: Forty consecutive successful transplantations were performed. Neither anastomosis leakage nor vessel obstruction occurred. The total operation time was 40-50 minutes. The time of cuff vascular anastomosis was 2-3 minutes and that of microvascular anastomosis was 9-12 minutes. All recipients survived for more than 30 days and all allografts were examined at 30 days after the transplantation. Pathological manifestations of allograft vessels were chronic rejection. CONCLUSION: This modified model of cervical heterotopic cardiac transplantation is simple, practical and highly reproducible and is applicable for investigation of chronic rejection in various organ transplantation studies.

[Isografts on subsequent ischemia-reperfusion injury: experiment with rats].

OBJECTIVE: To investigate the feasibility of ex vivo adenovirus-mediated gene transfer of human interleukin10 (hIL10) via the pulmonary vein into lung isografts, and to investigate the effect of hIL-10 gene transfer on subsequent ischemia-reperfusion injury (IRI). METHODS: Fifty-six male SD rats were randomly divided into 4 equal groups: Group D, undergoing left lung isotransplantation with the improved cuff anastomosis technique (the Isografts were transvascularly transfected 5 ml of 5 x 10(9) plaque-forming units/ml adenovec-hIL-10 complex, Group C, with the Isografts transvascularly transfected with blank adenovirus vector Adenovec, Group B, with the Isografts transvascularly transfected with diluent , and Group A, undergoing sham operation. All allografts were preserved for 3 hours at 10 degrees C before transplantation. Four hours after reperfusion blood samples were collected from hr abdominal aorta to undergo blood air analysis. Lung function was evaluated by partial pressure of oxygen (PaO2). Then the rats were killed with their left lung taken out to undergo pathological examination. The graft lung wet-to-dry (W/D) weight ratio was measured. SABC immunohistochemistry was used to detect the expression of hIL-10 in the cytoplasm. ELISA was used to detect the expression of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). The levels of malonyldialdehyde (MDA), superoxide dismutase (SOD), and myeloperoxidase (MPO) were measured by. Pathological morphologic change was also analyzed. RESULTS: The PaO2 level of Group D was significantly higher than those of Groups B and C (both P < 0.01). The W/D ratio, and levels of MDA and MPO of Group D were significantly lower than those of Groups B and C (both P < 0.01), but the SOD level of Group D was significantly higher than those of Groups B and C (both P < 0.05). The TNF-alpha and IFN-gamma levels of Group D were significantly lower than those of Groups B and C (both P < 0.01). Fewer tissue edema and interstitial inflammation were found in lungs. Of Group D RT-PCR showed hIL-10 expression in the lungs of the rats of Group D, but not in other groups. CONCLUSION: Ex vivo adenovirus-mediated gene transfer of hIL-10 via the pulmonary vein into the lung isografts is feasible and effective. hIL-10 gene transfer into lung isografts ameliorates subsequent IRI and improves early posttransplant graft function.