Local infiltration of HYR-PB21, a sustained-release formulation of bupivacaine, provides analgesia and reduces opioid requirement after haemorrhoidectomy: a randomised controlled trial.

BACKGROUND: HYR-PB21 is a new sustained-release formulation of bupivacaine indicated for controlling postoperative pain. The objectives of this study were to investigate the analgesic efficacy and safety profile of HYR-PB21 in patients after haemorrhoidectomy. METHODS: This was a multicentre, randomised, double-blind, positive-controlled trial. Patients were assigned randomly to receive a single dose of HYR-PB21 (150 mg or 300 mg) or bupivacaine HCl (75 mg) after surgery for prolapsing haemorrhoids. Postoperative pain was evaluated using a numeric rating scale at rest to calculate a cumulative pain score. Total rescue opioid usage and the proportion of subjects receiving rescue opioid were also assessed. RESULTS: We enrolled 72 patients with haemorrhoidectomy, and 71 patients completed the study. The average cumulative pain score through 72 h after surgery in the 300 mg HYR-PB21 group (87 scores) was lower than in the bupivacaine HCl group (166 scores) in an intention-to-treat analysis (P<0.001). There was a dose-response effect in reducing total opioid usage and the proportion of rescue opioid use between the 150 mg and 300 mg HYR-PB21 groups, with bupivacaine HCl as a reference group. The HYR-PB21 groups did not show more adverse effects than the bupivacaine HCl group. CONCLUSIONS: Local infiltration of a single dose of HYR-PB21 sustained-release bupivacaine had better efficacy in controlling postoperative pain, with similar adverse effects, compared with a single dose of bupivacaine HCl in patients after haemorrhoidectomy. CLINICAL TRIAL REGISTRATION: ChiCTR2000041318 (Chinese Clinical Trial Registry).

Surgical repair of a special category of arteriovenous fistula outflow stenosis caused by venous valve hyperplasia.

OBJECTIVE: This study evaluated a special category of arteriovenous fistula outflow stenosis caused by venous valve hyperplasia and explored the effectiveness of surgical repair in dealing with this kind of stenosis. STUDY DESIGN: This retrospective cohort study was conducted from February 2016 to January 2020 in our center. Patients with arteriovenous fistula dysfunction, including flow rate insufficiency, venous hypertension, thrombosis, and aneurysm dilation enlargement, were selected. Stenosis lesions presenting with venous valve hyperplasia were selected after ultrasound screening. All patients underwent surgical repair and were followed up every 6 months after surgery. RESULTS: Forty-three patients (median age, 54.5 ± 11.2 years; 65.1% men) were included. All procedures were technically successful. Based on intraoperative exploration, 56.5% were reconstructed via autologous vein patch, 17.4% of patients were reconstructed with end-to-end reconstruction after cutting the stenotic segment, 13.0% of cases simply had the valve resected, and 13.0% of cases involved a longitudinal incision and transverse suture. All patients returned to routine dialysis the following day and avoided catheter insertion. The mean follow-up time was 22.5 ± 14.0 (range, 1.3-49.8) months. The patency rates at 2 and 4 years were 92.2% and 79.0%, respectively. Valves harvested from patients were analyzed via Masson staining and immunohistochemical staining, indicating collagen fiber and myofibroblast hyperplasia in outflow venous valve hyperplasia (OVVH). CONCLUSIONS: Outflow venous valve hyperplasia can lead to fistula dysfunction. Ultrasound is the main method to diagnosis OVVH. Special surgical repair can preserve valuable vascular resources and relieve stenosis, is safe and effective, and has a high patency rate.

Long-term patency and comparisons of venous outflow in hemodialysis forearm arteriovenous grafts.

INTRODUCTION: This retrospective study investigated the factors and the effects of different venous outflows on forearm arteriovenous graft patency. METHODS: The venous outflow sites included basilic, cephalic, median antecubital, and deep veins. Comparisons among multiple groups were analyzed. FINDINGS: A total of 179 patients with forearm loop arteriovenous grafts met the inclusion criteria. Of these, 72 were basilic, 48 were cephalic, 44 were median antecubital, and 15 were deep. The median observation period was 19 months. The survival rate was 84.9% at 24 months and 78.2% at 48 months. Primary, secondary, and assisted primary patency rates for all arteriovenous grafts were 48.9%, 72.4%, and 68.4% at 12 months; 13.8%, 33.9%, and 23.6% at 24 months; and 0.6%, 4.6%, and 2.3% at 48 months, respectively. Differences in primary patency were statistically significant compared with those of secondary and assisted primary patency (P < 0.05). Primary patency rates for cephalic, median antecubital, basilic, and deep were 47.9%, 48.6%, 47.7%, and 40.0% at 12 months and 12.5%, 13.9%, 22.7%, and 0% at 24 months, respectively. Secondary patency rates for cephalic, median antecubital, basilic, and deep were 75.0%, 69.4%, 75.0%, and 73.3% at 12 months and 39.6%, 30.6%, 38.6%, and 13.3% at 24 months, respectively. There was no significant difference in primary thrombosis among basilic, cephalic, median antecubital and deep. There were no significant differences observed in primary or secondary patency rates among all the groups. Stenoses in the venous anastomosis and outflow vein were frequently observed in all types of arteriovenous grafts. Central venous stenosis was most commonly seen in deep (26.67%). On average, 1.9 interventions per patient were performed on the graft to maintain function. CONCLUSION: Different venous outflow selections were not associated with long-term patency and the occurrence of thrombosis in hemodialysis forearm loop arteriovenous grafts.

Efficacy and Safety of Paclitaxel-Coated Balloon Angioplasty for Dysfunctional Arteriovenous Fistulas: A Multicenter Randomized Controlled Trial.

RATIONALE & OBJECTIVE: Previous studies have illustrated the potential superiority of drug-coated balloons (DCBs) in maintaining patency after initial angioplasty for arteriovenous fistula (AVF) dysfunction due to stenosis. Our trial evaluated the efficacy and safety of DCBs for preventing fistula restenosis in Chinese hemodialysis patients. STUDY DESIGN: Multicenter, prospective, randomized, open-label, blinded end point, controlled trial. SETTINGS & PARTICIPANTS: A total of 161 hemodialysis patients with fistula dysfunction from 10 centers in China. INTERVENTION: Participants were randomized 1:1 to treatment with initial dilation followed by DCB angioplasty or conventional high-pressure balloon (HPB) angioplasty. OUTCOMES: The primary end point was target lesion primary patency defined as the target lesion intervention-free survival in conjunction with an ultrasonography-measured peak systolic velocity ratio (PSVR) ≤2.0 at 6 months. The secondary end points included 1) device, technical, clinical, and procedural success; 2) major adverse events; 3) degree of target lesion stenosis at 6 months; and 4) clinically driven target lesion and target shunt revascularization within 12 months. RESULTS: The percentage with target lesion primary patency as defined by a PSVR ≤2.0 was higher in the DCB group than in the control group (65% vs 37%, respectively; rate difference, 28% [95% CI, 13%-43%]; P <0.001) at 6 months. The target lesion and target shunt intervention-free survival of the DCB group were not superior to those of the control group at 6 months (P = 0.3 and P = 0.2, respectively) but were superior at 12 months (target lesion intervention-free survival: 73% for DCB vs 58% for control [P = 0.04]; target shunt intervention-free survival: 73% for DCB vs 57% for control [P = 0.04]). The average degree of target lesion stenoses at 6 months was not significantly different between the 2 groups (44% ± 16% for DCB vs 49% ± 18% for control; P = 0.09). There were no significant differences in major adverse events or in device, technical, clinical, or procedural success rates between the groups. LIMITATIONS: Small sample size; short follow-up period; procedural differences between the 2 groups such as unequal inflation times and balloon lengths. CONCLUSIONS: Compared to conventional HPB angioplasty, DCB treatment achieved superior primary patency defined using PSVR measured at 6 months and superior intervention-free survival of both the target lesion and the target shunt at 12 months without evidence of greater adverse events. FUNDING: Funded by ZhuHai Cardionovum Medical Device Co., Ltd. TRIAL REGISTRATION: Registered at ClinicalTrials.gov with study number NCT02962141.

Sepsis-Induced Thymic Atrophy Is Associated with Defects in Early Lymphopoiesis.

Impaired T lymphopoiesis is associated with immunosuppression of the adaptive immune response and plays a role in the morbidity and mortality of patients and animal models of sepsis. Although previous studies examined several intrathymic mechanisms that negatively affect T lymphopoiesis, the extrathymic mechanisms remain poorly understood. Here, we report a dramatic decrease in the percentage of early T lineage progenitors (ETPs) in three models of sepsis in mice (cecal ligation and puncture, lipopolysaccharide continuous injection, and poly I:C continuous injection). However, septic mice did not show a decrease in the number of bone marrow (BM) precursor cells. Instead, the BM progenitors for ETPs expressed reduced mRNA levels of CC chemokine receptor (CCR) 7, CCR9 and P-selectin glycoprotein ligand 1, and exhibited impaired homing capacity in vitro and in vivo. Furthermore, RNA-Seq analysis and real-time PCR showed a marked downregulation of several lymphoid-related genes in hematopoietic stem and progenitor cells. Hematopoietic stem and progenitor cells differentiated into myeloid cells but failed to generate T lymphocytes in vitro and in vivo. Our results indicate that the depletion of ETPs in septic mice might be a consequence of an impaired migration of BM progenitors to the thymus, as well as a defect in lymphoid lineage commitment. Stem Cells 2016;34:2902-2915.

Arterial variations and hemodynamic impact in the upper limb: Insights from an observational study.

BACKGROUND: The incidence of arterial variations in the upper limb is variable among ethnic groups. However, there are only a few studies on the Asian population, particularly in China. OBJECTIVE: This research provided the first large data series on arterial variations in the upper limb of Chinese adults. METHODS: In this observational study, the arteries in the upper limb of consecutive patients were analyzed between July 2020 and June 2021, and the hemodynamic variables of each patient were collected. RESULTS: The brachial artery deviated in 6.60% of upper limbs (11.89% of patients), and the brachioradial artery (BRA) was the most frequent variation. In the wrists of all patients, the volume flow (VF) of the BRA was lower than that of the radial artery (RA). Besides, the VF of the BRA in patients with end-stage renal disease (ESRD) was lower than that in those without ESRD in the wrist. Additionally, in the forearms of patients without ESRD, the VF of the BRA was lower than that of the RA. In the upper arm of patients with ESRD, the VF of the brachial artery in patients with BRA was significantly lower than that in patients with RA. CONCLUSION: The incidence of arterial variations in the upper limb of Chinese adults was 6.60% (limbs) or 11.89% (patients) in this study. In our investigation, the BRA was the most common high bifurcation of the brachial artery type. Furthermore, the Chi-squared test revealed significant differences in the distribution, which was common in the right upper limb but not in sex.

Drug-Coated Balloon Angioplasty for Dysfunctional Arteriovenous Hemodialysis Fistulae: A Randomized Controlled Trial.

BACKGROUND: The aim of this study was to evaluate the efficacy and safety of paclitaxel-coated balloons (AcoArt Orchid) in treating dysfunctional arteriovenous fistulae. METHODS: The drug-eluting balloon for arteriovenous (AV) fistula in China trial was a prospective, multicenter, randomized controlled study. Patients who had ≥50% venous stenosis of the AV fistula and symptoms indicating significant hemodynamic changes were included. After successful predilation with a high-pressure balloon (residual stenosis ≤30%), patients were randomized 1:1 to either a paclitaxel-coated balloon or an uncoated control balloon. The primary efficacy outcome was assessed at 6 months, and safety assessment was conducted within 30 days of the procedure. The 12-month results were also analyzed. RESULTS: The study included 244 patients, equally distributed between the two groups. The primary target lesion patency was 91% (106/116) for the drug-coated balloon (DCB) group and 67% (79/118) for the plain balloon catheter group, representing a difference of 24.63% (95% confidence interval, 14.68 to 34.58; P < 0.001). The secondary efficacy end point was primary target lesion patency at 12 months, which was 66% (74/112) for the DCB group and 46% (52/112) for the plain balloon catheter group (95% confidence interval, 6.57 to 32.08; P = 0.004). The mean number of reinterventions per patient to maintain target lesion patency during the 12 months after the index procedure was 0.39 (48/122) in the DCB group and 0.77 (94/122) in the plain balloon catheter group ( P = 0.001). The primary safety end point did not differ between groups ( P = 0.25). CONCLUSIONS: AcoArt Orchid DCB showed better primary patency rates compared with plain balloon angioplasty for treating stenotic lesions in dysfunctional hemodialysis AV fistulae at 6 and 12 months. It required fewer repeated interventions and had comparable safety in 1 year. CLINICAL TRIAL REGISTRY NAME AND REGISTRATION NUMBER: AcoArt III/Arterio-venous Fistula in China, NCT03366727 .

Hypothermia predicts the prognosis in colon ascendens stent peritonitis mice.

BACKGROUND: The mortality rate of severe sepsis remains unacceptably high. It is difficult to make advances in the treatment of this problematic and increasingly frequent medical condition. In severe sepsis, hypothermia can be recognized as an important feature. The present study investigated the role of hypothermia in the prognosis of the colon ascendens stent peritonitis (CASP) model. METHODS: We employed the CASP model for wild-type C57BL/6 mice. We compared physiologic indices in survivor and non-survivor groups after CASP to test whether low temperature might be a helpful predictor in sepsis. To certify this hypothesis, we examined the survival rate, peritoneal leukocytes, and organ damage. We also measured the bacterial burden and inflammatory cytokine levels at different times. RESULTS: The temperature varied dramatically in the survivors' group compared with the non-survivors' group at 18 h. We divided the CASP models into a mild group and a severe group, based on temperatures above or below 32°C at 18 h. Mice in the severe group had a lower survival rate (0% versus 87.5%), more peritoneal leukocytes, more bacterial culture results, higher expressions of cytokines, and more classical features in pathology compared with the mild group. CONCLUSIONS: Hypothermia (below 32°C at 18 h) might be a predictor of prognosis in CASP-induced sepsis.

Resection of Carotid Body Tumors in Patients of Advanced Age: Experience From a Single Center.

INTRODUCTION: Resection of carotid body tumor (CBT) in patients of advanced ages has not been appreciated. OBJECTIVES: This study aims to assess the clinical characteristics and perioperative comorbidities for CBT resection in patients of advanced age and to validate the application of an "isolated island" technique for extirpation of CBT. METHODS: Eight patients of advanced age (≥60 years) who underwent CBT resection were enrolled as the study group (SG). Another 29 patients of younger age (<45 years old) underwent CBT extirpation were assigned as the control group (CG). The perioperative issues were compared between these 2 groups. RESULTS: The "isolated island" technique was successfully applied for resection of CBT in all 37 patients. The prevalence of Shamblin classification I, II, and III tumors in the SG was 12.5%, 62.5%, and 25%; whereas in the CG was 10.3%, 55.2%, and 34.5%, respectively. Bilateral CBT was observed in 7 patients of the CG and none in the SG. Vascular reconstruction was required for 1 (12.5%) patient in the SG, while it was required for 8 (27.6%) patients in the CG. Postoperative vocal cord palsy occurred in 37.5% of patients in SG, whereas the vocal cord palsy (34.5%) and dysphagia (6.9%) were commonly encountered in CG. In addition to postoperative length of stay (P = .004), no significant difference for operative time, intraoperative blood loss, or mortality were observed between these 2 groups (P > .05). CONCLUSION: Extirpation of CBT in patients of advanced age is rationale in appropriately selected patients. The "isolated island" technique is safe for CBT resection with seemingly low complication rates.

Sepsis induces non-classic innate immune memory in granulocytes.

Secondary infection in patients with sepsis triggers a new wave of inflammatory response, which aggravates organ injury and increases mortality. Trained immunity boosts a potent and nonspecific response to the secondary challenge and has been considered beneficial for the host. Here, using a murine model of polymicrobial infection, we find that the primary infection reprograms granulocytes to boost enhanced inflammatory responses to the secondary infection, including the excessive production of inflammatory cytokines, respiratory burst, and augmented phagocytosis capacity. However, these reprogramed granulocytes exhibit "non-classic" characteristics of innate immune memory. Two mechanisms are independently involved in the innate immune memory of granulocytes: a metabolic shift in favor of glycolysis and fatty acid synthesis and chromatin remodeling leading to the transcriptional inactivity of genes encoding inhibitors of TLR4-initiated signaling pathways. Counteracting the deleterious effects of stressed granulocytes on anti-infection immunity might provide a strategy to fight secondary infections during sepsis.

[Surgical management of penetrating cervical vascular trauma].

OBJECTIVES: To discuss diagnosis and surgical management of penetrating cervical vascular trauma. METHODS: A retrospective clinical analysis of 22 penetrating carotid artery injuries. RESULTS: Twenty-two patients presented 32 vascular injuries, including innominate artery (n = 1), innominate vein (n = 2), subclavian artery (n = 6), subclavian vein (n = 2), common carotid artery (n = 3), internal carotid artery (n = 3), external carotid artery (n = 4), jugular vein (n = 8) and vertebral artery (n = 4). There were 12 patients with stab wounds, 2 with blast wound and 8 with iatrogenic injuries. Of these, there were 12 zone-1 injuries (38%), 19 zone-2 injuries (59%) and 1 zone-3 injury (3%). The distribution of 4 vertebral artery injuries were V1 (n = 1), V2 (n = 2) and V3 (n = 1). All patient received surgical and endovascular managements and got survival. CONCLUSIONS: Patients with penetrating cervical vascular injuries have high rate of mortality. Emergent surgical exploration is necessary for patients with hard signs of vascular injury such as hemodynamic instability, exsanguinating hemorrhage, or expanding hematoma. Those patients that are hemodynamically stable and who are without respiratory compromise should undergo further diagnostic imaging evaluation.

TRIM21­a potential biomarker for the prognosis of thyroid cancer.

Thyroid cancer (THCA) is one of the commonest malignancies associated with increased recurrence. Therefore, identifying the putative molecular markers and therapeutic targets to improve the treatment of THCA is essential. The present study analyzed the potential role of tripartite motif-containing 21 (TRIM21), a member of the TRIM family belonging to the subfamily of E3 ubiquitin ligases, in the progression of THCA. Using bioinformatics analysis and immunohistochemistry of THCA tissues, it was observed that TRIM21 is overexpressed in THCA tissues. The present study also found that TRIM21 is associated with lymph node metastasis and high-risk recurrence of THCA. Furthermore, it identified a promotional role of TRIM21 in THCA cell migration and invasion. In addition, the present study analyzed TRIM21-enriched pathways and co-expressed genes in THCA. The present study suggested that TRIM21 may serve as a potential biomarker for THCA prognosis.

Acellular Vascular Scaffolds Preloaded With Heparin and Hepatocyte Growth Factor for Small-Diameter Vascular Grafts Might Inhibit Intimal Hyperplasia.

To develop small-diameter (<6 mm) scaffolds capable of accelerating rapid endothelialization and improving long-term patency rate, we created acellular vascular scaffolds preloaded with heparin and hepatocyte growth factor (HGF). Heparin was conjugated to suppress thrombogenic responses, and HGF was immobilized to induce endothelial cells (ECs) proliferation and migration. The scaffolds immobilized with heparin exhibited highly effective localization and sustained release of HGF for 30 days in vitro. We implanted this modified scaffold into the carotid artery of a rabbit model to investigate the efficacy in vivo. The acellular vascular scaffold with heparin only was used as control. After transplantation, the patency of this modified scaffold was 91.67% at 1, 3, 6, and 12 months, while the patency rate in the group with grafted heparin only was 83.33% at 1, 3, 6, and 12 months. This modified scaffold significantly stimulated ECs proliferation and the endothelium aligned in the direction of flow after 12 months. In addition, intimal hyperplasia was significantly reduced in the grafts coated with HGF compared with the control grafts. The small-diameter vascular grafts with an inner diameter of 2.5 mm preloaded with heparin and HGF may be a substitute for autologous blood vessels in clinic.

Mutant prominin 1 found in patients with macular degeneration disrupts photoreceptor disk morphogenesis in mice.

Familial macular degeneration is a clinically and genetically heterogeneous group of disorders characterized by progressive central vision loss. Here we show that an R373C missense mutation in the prominin 1 gene (PROM1) causes 3 forms of autosomal-dominant macular degeneration. In transgenic mice expressing R373C mutant human PROM1, both mutant and endogenous PROM1 were found throughout the layers of the photoreceptors, rather than at the base of the photoreceptor outer segments, where PROM1 is normally localized. Moreover, the outer segment disk membranes were greatly overgrown and misoriented, indicating defective disk morphogenesis. Immunoprecipitation studies showed that PROM1 interacted with protocadherin 21 (PCDH21), a photoreceptor-specific cadherin, and with actin filaments, both of which play critical roles in disk membrane morphogenesis. Collectively, our results identify what we believe to be a novel complex involved in photoreceptor disk morphogenesis and indicate a possible role for PROM1 and PCDH21 in macular degeneration.

Microbiological profile and clinical characteristics of diabetic foot infection in northern China: a retrospective multicentre survey in the Beijing area.

PURPOSE: We aimed to define the microbiological characteristics of diabetic foot infection in patients in the Beijing area and to explore the demographic and clinical factors correlated with pathogen distribution. METHODOLOGY: As part of a retrospective multicentre surveillance program conducted in eight hospitals in Beijing 2010-2014, we recruited all inpatients for whom bacterial culture had been performed. Demographic, clinical, laboratory and surgery data were obtained from medical records. Statistical analysis was performed to analyse data on microbiological and clinical characteristics.Results/Key findings. A total of 456 cases were included. The culture positivity was 95.4 %. Among all patients with positive cultures, 88 cases (20.2 %) had polymicrobial infections. Five hundred and fifty-one species were isolated from all specimens, including 39.6 % Gram-positive bacteria and 57.5 % Gram-negative bacteria. Enterobacteriaceae accounted for 41.0 % of all isolates. Staphylococcus aureus (17.1 %), Pseudomonas aeruginosa (13.1 %), Proteus spp. (9.8 %), Escherichia coli (9.3 %) and coagulase-negative Staphylococcus (8.3 %) were the most frequently isolated species. The rate of resistance to methicillin was 24.5 % for S. aureus. The susceptibility of P. aeruginosa to all antibiotics was over 60 %. The rate of extended-spectrum ß-lactamase production among E. coli was 52.6 %. P. aeruginosa and Enterobacteriaceae show high sensitivity to piperacillin/tazobactam, carbapenems and amikacin. Multivariate analysis showed that patient age >60 years was independently associated with Gram-negative rods. CONCLUSIONS: Enterobacteriaceae were the most frequently isolated organisms in our area. Older patients were more likely to suffer from Gram-negative rod infections. Gram-negative rods show high sensitivity to piperacillin/tazobactam, carbapenems and amikacin.

Increased levels of interleukin-10 in serum from patients with hepatocellular carcinoma correlate with profound numerical deficiencies and immature phenotype of circulating dendritic cell subsets.

Increased levels of interleukin (IL)-10 have been described as a negative prognostic indicator for survival in patients with various types of cancer. IL-10 exerts tolerogenic and immunosuppressive effects on dendritic cells, which are crucial for the induction of an antitumor immune response. Blood dendritic cell antigen (BDCA)-2 and BDCA-4 are specifically expressed by CD123(bright) CD11c- plasmacytoid dendritic cells; whereas BDCA-1 and BDCA-3 define 2 distinct subsets of CD11c+ myeloid dendritic cells. In this study, the T-helper cell (Th)1/Th2 cytokine serum profile of 65 hepatocellular carcinoma patients was assessed. We found that serum levels of IL-10 were substantially increased in hepatocellular carcinoma patients as compared with controls. Peripheral blood mononuclear cells from healthy volunteers were exposed to recombinant human (rh)IL-10 in vitro to additionally characterize its impact on distinct blood dendritic cell subsets. A dramatic decrease of all myeloid dendritic cell (MDC) and plasmacytoid dendritic cell (PDC) subsets was detectable after 24 hours of continuous rhIL-10 exposure. Moreover, the expression of HLA-DR, CD80 and CD86, was significantly reduced on rhIL-10-treated dendritic cell subsets. Direct ex vivo flow cytometric analysis of various dendritic cell subpopulations in peripheral blood from hepatocellular carcinoma patients revealed an immature phenotype and a substantial reduction of circulating dendritic cells that was associated with increased IL-10 concentrations in serum and with tumor progression. These findings confirm a predominantly immunosuppressive role of IL-10 for circulating dendritic cells in patients with hepatocellular carcinoma and, thus, may indicate novel aspects of tumor immune evasion.

A novel RDS/peripherin gene mutation associated with diverse macular phenotypes.

Pattern dystrophy is a heterogeneous group of retinal dystrophies of which butterfly-shaped pattern dystrophy (BPD) and adult-onset foveomacular dystrophy (AOFMD) are the two most common forms. BPD is characterized by a butterfly-shaped, irregular, depigmented lesion at the level of the retinal pigment epithelium. In contrast, AOFMD is characterized by the presence of slightly elevated, symmetric, solitary, round to oval, yellow lesions at the level of the retinal pigment epithelium. We identified three independent kindreds with pattern dystrophy, one with four patients affected with BPD and the other two with 14 affected patients with AOFMD. We performed complete ophthalmic examination, fluorescein angiography, linkage mapping, and mutational screening in the RDS/peripherin gene in the affected patients. Patients affected with BPD had a best-corrected vision of 20/20 to 20/25, whereas vision in the eyes of patients with AOFMD ranged from 20/20 to 20/400. In all three kindreds, sequence analysis identified an A-to-G change at nucleotide position 422 of the RDS/peripherin gene, predicting a novel Tyr-141-Cys substitution. A haplotype analysis revealed that these three kindreds shared an identical disease haplotype at the RDS/peripherin locus, indicating that the mutation reflects a founder effect. The sequence change that segregated with the disease phenotype was not observed in 200 control chromosomes. Our results identified a novel mutation in the RDS/ peripherin gene that can cause diverse macular phenotypes. Genetic and clinical investigation of pattern dystrophy may provide useful diagnostic tools and new treatment strategies for this disorder.

[Effect of ischemic pretreatment on expression of heat shock protein 70 and injury of spinal cord in canine].

OBJECTIVE: To evaluate effect of ischemic pretreatment on expression of heat shock protein 70 (HSP70) and injury of spinal cord in canine. METHODS: Fourty-one canine were divided into three groups: the sham-operative group, the pretreatment group and the control group. In the pretreatment group aorta was obstructed for 6 min, and then was opened for 6 min, this procedure was repeated twice, finally aorta was obstructed for 35 min. In the control group aorta was obstructed for 35 min. Nervous function were assessed and HSP70 expression were detected in tissue of spinal cord. RESULTS: In the pretreatment group, HSP70 expressed in cytoplasm and nucleus at 6, 24 hour after reperfusion, and intensity of HSP70 expression was stronger than that in the control group; The score of nervous function in the pretreatment group was higher than that in the control group. On 7 day after reperfusion the score of nervous function in pretreatment group had no obvious variation, and HSP70 expression was still observed. CONCLUSIONS: Ischemic pretreatment can improve ischemic tolerance of spinal cord; HSP70 expression in cytoplasm and nucleus may play a role in ischemic tolerance.

Identification of a novel VEGFR-3 missense mutation in a Chinese family with hereditary lymphedema type I.

A novel mutation of vascular endothelial growth factor receptor gene (VEGFR-3), was identified in a four-generation Chinese family with hereditary lymphedema type I (HL-I). Genetic linkage analysis was performed on the known genetic locus for HL-I with a panel of polymorphic markers, and then mutations were screened out by direct sequencing. By genotyping, the family showed the linkage to HL-I locus on 5q35.3. Mutation screening analysis of the exons encoding the intracellular kinase domains of VEGFR-3, revealed a novel missense mutation D1055V. This mutation cosegregated with the disease phenotype in the family and was not found in 100 normal controls. This finding has expanded the spectrum of the VEGFR-3 gene mutations causing HL-I, and will be useful for further genetic consultation and genetic diagnosis.

Increased frequencies of CD8+ T lymphocytes recognizing wild-type p53-derived epitopes in peripheral blood correlate with presence of epitope loss tumor variants in patients with hepatocellular carcinoma.

Wild-type (WT) sequence p53 peptides are attractive candidates for broadly applicable cancer vaccines. The aim of this study was to evaluate the potential of a WT p53-based immunotherapeutic approach for patients with hepatocellular carcinoma (HCC). Circulating CD8+ T cells specific for WT p53(149-157) and WT p53(264-272) HLA-A*0201 restricted epitopes were directly identified in the peripheral blood by the use of peptide/HLA-A2.1 tetramers in 24 HCC patients. Cytotoxic T lymphocyte (CTL) activity after WT p53 peptide-specific stimulation was assessed by analysis of granzyme B and interferon-gamma mRNA transcription, using a quantitative real-time polymerase chain reaction assay. Tumor immunophenotyping was performed to evaluate the p53 status, the expression of major histocompatibility complex (MHC) and costimulatory molecules in freshly isolated tumor cells. HCC patients exhibited significantly higher frequencies of WT p53-specific memory CD8+ T cells and stronger WT p53-specific CTL activity, when compared with healthy controls. Increased frequencies of p53-specific CD8+ T cells and their activity correlated with selective HLA-A2 allele loss and reduced costimulatory molecule expression of tumor cells. Moreover, augmented numbers of p53-specific T cells coincided with high MHC class II expression in tumor cells but were inversely related to the T status of the tumor node metastasis staging system. Our results indicate the existence of natural immunosurveillance and tumor immune evasion, involving a T cell response against WT p53 tumor antigen in patients with HCC. These findings may have important implications for the future development of cancer vaccines.