[Multicenter real world study on the efficacy and safety of eribulin for the treatment of advanced breast cancer].

Objective: To explore the efficacy and safety of real-world eribulin in the treatment of metastatic breast cancer. Methods: From December 2019 to December 2020, patients with advanced breast cancer were selected from Beijing Chaoyang District Sanhuan Cancer Hospital, Shandong Cancer Hospital, Peking University Cancer Hospital, Baotou Cancer Hospital, Shengjing Hospital Affiliated to China Medical University, and Cancer Hospital of Chinese Academy of Medical Sciences. Kaplan-Meier method and Log rank test were used for survival analysis, and Cox regression model was used for multivariate analysis. Results: The median progression-free survival (PFS) of 77 patients was 5 months, the objective response rate (ORR) was 33.8%, and the disease control rate (DCR) was 71.4%. The ORR of patients with triple-negative breast cancer was 23.1%, and the DCR was 57.7%; the ORR of patients with Luminal breast cancer was 40.0%, and the DCR was 77.8%; the ORR of patients with HER-2 overexpression breast cancer was 33.3%, and the DCR was 83.3%. ORR of 50.0% and DCR of 66.7% for patients treated with eribulin as first to second line treatment, ORR of 29.4% and DCR of 76.5% for patients treated with third to fourth line and ORR of 28.6% and DCR of 71.4% for patients treated with five to eleven line. The ORR of patients in the eribulin monotherapy group was 40.0% and the DCR was 66.0%; the ORR of patients in the combination chemotherapy or targeted therapy group was 22.2% and the DCR was 81.5%. Patients with a history of treatment with paclitaxel, docetaxel, or albumin paclitaxel during the adjuvant phase or after recurrent metastasis had an ORR of 32.9% and a DCR of 69.9% when treated with eribulin. The treatment efficacy is an independent prognostic factor affecting patient survival (P<0.001). The main adverse reactions in the whole group of patients were Grade Ⅲ-Ⅳ neutrophil decline [29.9% (23/77)], and other adverse reactions were Grade Ⅲ-Ⅳ fatigue [5.2% (4/77)], Grade Ⅲ-Ⅳ peripheral nerve abnormality [2.6% (2/77)] and Grade Ⅲ-Ⅳ alopecia [2.6% (2/77)]. Conclusions: Eribulin still has good antitumor activity against various molecular subtypes of breast cancer and advanced breast cancer that has failed multiple lines of chemotherapy, and the adverse effects can be controlled, so it has a good clinical application value.

[Phase â ¢ randomized controlled, multicenter, prospective study of recombinant anti-HER2 humanized monoclonal antibody (Cipterbin) combined with vinorelbine in patients with HER2 positive metastatic breast cancer: the HOPES Study].

Objective: To evaluate the clinical efficacy and safety of recombinant anti-HER2 humanized monoclonal antibody (Cipterbin) combined with vinorelbine in patients with HER2 positive metastatic breast cancer. Methods: Patients were randomized 2∶1 to test group and control group. Patients in test group received Cipterbin (4 mg/kg loading dose and 2 mg/kg maintenance dose each week, IV) combined with vinorelbine (25 mg/m(2) on days 1,8 and 15 of each 28 days, IV). Patients in control group received vinorelbine (25 mg/m(2) on days 1,8 and 15 of each 28 days, IV).The primary end point was progression free survival (PFS). Results: A total of 315 patients were enrolled from Jan 2009 to Jan 2013 (212 in test group and 103 in control group). The median PFS of test group was significantly longer than that of control group, 39.1 weeks vs 14.0 weeks (HR=0.24; 95%CI, 0.16-0.36; P<0.000 1). The objective response rate (ORR) and disease control rate (DCR) in test group were significantly higher than those in control group, ORR was 46.7% vs 18.45% (P<0.000 1) and DCR was 79.72% vs 45.63% (P<0.000 1). The incidence of neutropenia, leucopenia and erythrocytopenia were higher in both groups, but there was no significant difference between two groups.The most common adverse events associated with Cipterbin were infusion reactions. Left ventricular ejection fraction reduced to less than 50% in 5 patients, which were recovered. No serious cardiotoxicity. Conclusion: The recombinant anti-HER2 humanized monoclonal antibody (Cipterbin) combined with vinorelbine has significant efficacy and good safety. It is the optimized therapy regime for patients with taxane-pretreated HER2 positive metastatic breast cancer, which provides more targeted therapy opportunities for HER2 positive breast cancer patients in China.

Transplantation of Skin Precursor-Derived Schwann Cells Yields Better Locomotor Outcomes and Reduces Bladder Pathology in Rats with Chronic Spinal Cord Injury.

Cell transplantation for spinal cord injury (SCI) has largely been studied in sub-acute settings within 1-2 weeks of injury. In contrast, here we transplanted skin-derived precursors differentiated into Schwann cells (SKP-SCs) into the contused rat spinal cord 8 weeks post-injury (wpi). Twenty-one weeks later (29 wpi), SKP-SCs were found to have survived transplantation, integrated with host tissue, and mitigated the formation of a dense glial scar. Furthermore, transplanted SKP-SCs filled much of the lesion sites and greatly enhanced the presence of endogenous SCs, which myelinated thousands of sprouting/spared host axons in and around the injury site. In addition, SKP-SC transplantation improved locomotor outcomes and decreased pathological thickening of bladder wall. To date, functional improvements have very rarely been observed with cell transplantation beyond the sub-acute stage of injury. Hence, these findings indicate that skin-derived SCs are a promising candidate cell type for the treatment of chronic SCI.

[Efficacy and safety of oral pyrotinib in HER2 positive metastatic breast cancer: real-world practice].

OBJECTIVE: Pyrotinib, a novel irreversible pan-ErbB receptor tyrosine kinase inhibitor, showed promising antitumor activity and acceptable tolerability in phase II and phase III randomized clinical trials. We assessed the activity and safety of oral pyrotinib for human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer patients in the real world. METHODS: We retrospectively analyzed 72 HER2 positive metastatic breast cancer (MBC) patients who received oral pyrotinib based regimens at Beijing Cancer Hospital and other four hospitals (Peking University First Hospital, China-Japan Friendship Hospital, General Hospital of PLA, Peking University Third Hospital) from August 2018 to September 2019. Progression free survival (PFS), objective response rate (ORR), adverse events (AE) of pyrotinib were investigated. RESULTS: Seventy-two patients with HER2 positive MBC were enrolled. The median age of the patients was 55 years (range: 32-79 years). Sixty-nine (95.8%) patients had received anti-HER2 treatment in the metastatic and/or (neo) adjuvant settings; 61 (84.7%) patients had received anti-HER2 treatments in the metastatic setting in terms of trastuzumab 56 (77.8%) patients, lapatinib 36 (50.0%) patients, and T-DM1 4 (5.6%) patients. Among these 72 patients who received oral pyrotinib based regimens, 62 (86.1%) patients received pyrotinib (±trastuzumab) in combination with chemotherapy, 6 (8.3%) patients received pyrotinib (± trastuzumab) in combination with endocrine therapy and 4 (5.6%) patients received pyrotinib (±trastuzumab). Sixty-five (90.3%) patients received 400 mg pyrotinib once daily as initial dose, and 7 (9.7%) patients received 320 mg. OBJECTIVE response and safety to pyrotinib based therapy were evaluable in all the 72 patients. One (1.4%) patient achieved complete response (CR), 18 (25.0%) patients achieved partial response (PR), 41 (56.9%) patients had stable disease (SD), and 12 (16.7%) patients had progressive disease (PD). The ORR (CR+PR) was 26.4% and the median PFS was 7.6 months (95%CI: 5.5-9.7 months). Among the 36 patients with prior lapatinib therapy, the median PFS was 7.9 months (95%CI: 4.1-11.7 months). Among the 15 patients with brain metastasis, the median PFS was 6.0 months (95%CI: 2.2-9.8 months). The main toxicities related to pyrotinib were diarrhea in 57 (79.2%) cases, and 48 (66.7%) cases with grade 1-2 as well as 9 (12.5%) cases with grade 3. CONCLUSION: Pyrotinib based therapy is an effective treatment for patients with HER2 positive MBC, including patients with lapatinib treatment failure and brain metastasis, and the toxicities can be tolerated.

Effect of tannins and cellulase on growth performance, nutrients digestibility, blood profiles, intestinal morphology and carcass characteristics in Hu sheep.

Objective: This study was conducted to evaluate the effects of tannins and cellulase on growth performance, nutrient digestibility, blood profiles, intestinal morphology and carcass characteristics in Hu sheep. Methods: A total of 48 three-month-old meat Hu sheep (25.05 ± 0.9 kg) were blocked based on body weight, and randomly allotted to 4 treatments with 3 replicates of 4 sheep each. The experiment lasted for 80 d, and dietary treatments were as follows: (1) CON, control diet; (2) TAN, CON + 0.1% tannins; (3) CEL, CON + 0.1% cellulase; (4) TAN+ CEL, CON + 0.1% tannins and 0.1% cellulase. Results: Compared with CON, CEL and TAN+CEL had greater (P<0.05) final birth weight (FBW) and average daily gain but lower (P<0.05) F/G, while FBW of TAN+CEL was lower (P<0.05) than that of CEL. The apparent total tract digestibility (ATTD) of DM in TAN, CEL and TAN+CEL groups were higher (P<0.05) than that in CON. CEL and TAN+CEL groups had greater (P<0.05) ATTD of CF compared with TAN and CON, while TAN group had lower (P<0.05) ATTD of CP than other treatments. TAN, CEL and TAN+CEL groups increased (P<0.05) serum globulin and alkaline phosphatase but decreased (P<0.05) A/G. Serum total protein was greatest for TAN+CEL, intermediate for TAN and CEL and least for CON (P<0.05). TAN+CEL group increased (P<0.05) dressing percentage compared with CON, while the backfat thickness of CEL was lower (P<0.05) than that of CON. The villus height of jejunum and ileum in CEL and TAN+CEL groups were greater (P<0.05) than that in CON, and the crypt depth and villus height: crypt depth of jejunum were increased (P<0.05) in TAN, CEL and TAN+CEL groups. Conclusion: The addition of tannins and cellulase together promoted nutrient digestion, liver protein synthesis and intestinal development and thus improved growth performance and carcass characteristics.

[Epidemiological investigation of allergic rhinitis in Ningxia].

Objective: To investigate the epidemiological characteristics of allergic rhinitis (AR) in Ningxia and to analyze its related factors. Methods: From March to September of 2013, a multi-stage and cluster sampling method was used to investigate the diagnosis and treatment of AR in Ningxia Area (3 years and above). Guidelines for diagnosis and treatment of allergic rhinitis (2009, Wuyishan) was used as the basis for the diagnosis of adult AR, while Guidelines for diagnosis and treatment of pediatric allergic rhinitis (2010, Chongqing) was used as the basis for children. SPSS 16.0 software was used to complete the statistical analysis. Results: The total number of questionnaires was 6 000, and the number of effective questionnaire was 5 236, the recovery rate was 87.27%. With 684 cases diagnosed of AR, the prevalence of AR in Ningxia was 13.06% (684/5 236), including 13.40% (325/2 425) of males, 12.77% (359/2 811) of females. The difference was not statistically significant (χ(2)=0.456, P>0.05). There was significant difference in the prevalence between Hui and Han [14.35% (452/3 150) vs 11.12% (232/2 086), χ(2)=11.51, P<0.05]. According to ARIA criteria, persistent AR was 27.63% (189/684), intermittent AR was 72.37% (495/684). The month with highest incidence of AR in Ningxia Area was September, accounting for 71.78% (491/684). The prevalence of urban population was 14.54%, with the prevalence of rural population was 11.90%, and the difference was significant between urban and rural residents (χ(2)=7.90, P<0.05). The age group with highest prevalence rate was 21~30 years old. The main inhalation allergens were mugwort (68.42%), weeds (58.48%) and ragweed (55.56%). The main dietary allergens were wheat flour (14.33%), peanut (13.74%) and walnut kernel (11.99%). The most common complication was allergic conjunctivitis [82.02% (561/684)]. Conclusion: The epidemiology of AR in Ningxia Area is preliminarily understood, which will provide the epidemiological evidence for the prevention and treatment of AR and the formulation of public health policy.

Passive drug permeation through membranes and cellular distribution.

Although often overlooked, passive mechanisms can lead to significant accumulation or restriction of drugs to intracellular sites of drug action. These mechanisms include lipoidal diffusion of ionized species and pH partitioning according to the electrochemical potential and to pH gradients that exist across subcellular compartments, respectively. These mechanisms are increasingly being exploited in the design of safe and effective drugs for the treatment of a wide variety of diseases. In this work, the authors review these efforts and the associated passive mechanisms of cellular drug permeation. A generic mathematical model of the cell is provided and used to illustrate concepts relevant to steady-state intracellular distribution. Finally, the authors review methods for estimating determinant parameters and measuring the net effect at the level of unbound intracellular drug concentrations.

[Predictive and prognostic value of monitoring lymphocyte subsets in peripheral blood before and after chemotherapy in patients with metastatic breast cancer].

OBJECTIVE: To detect the proportion of lymphocyte subsets in peripheral blood of the advanced breast cancer patients before and after chemotherapy with docetaxel and thiotepa, as well as the association between the proportion of peripheral blood lymphocyte subsets with the response rate and prognosis. METHODS: The proportions of lymphocyte subsets (CD3+ T cell, CD3+/CD4+ T cell, CD3+/CD8+ T cell, CD3-/CD16+56+ NK cell, CD3+/CD16+56+ T cell, CD19+ B cell, CD4+/CD25+ T cell, CD8+/CD28- T cell, CD8+/CD28+ T cell) in the peripheral blood of 86 patients were analyzed with flowcytometry before and after chemotherapy. The result was analyzed in combination with clinicopathological data. RESULTS: The proportion of regulatory T cells (Treg) after chemotherapy in the disease control patients decreased significantly compared with that of the progressive patients (P=0.034). The difference of the proportions of Treg before and after chemotherapy affected significantly the overall survival (OS). The OS of the patients with decreased proportion of Treg was significantly longer than that of the patients with increased proportion of Treg, which was 23.5 and 9.4 months respectively (P<0.05). CONCLUSION: The patients with decreased proportion of Treg after chemotherapy had higher response rate and better survival benefit.

Radiographic evaluation of short-stem press-fit total shoulder arthroplasty: short-term follow-up.

BACKGROUND: Humeral loosening is an uncommon etiology for revision shoulder arthroplasty. We aimed to evaluate the radiographic and clinical outcomes of a short-stem press-fit humeral component after primary total shoulder arthroplasty. METHODS: We reviewed our patient database, from January 2008 to December 2011, for primary total shoulder arthroplasties performed with a short-stem press-fit humeral component. Radiographs and clinical outcomes were evaluated in the immediate postoperative period and at the most recent follow-up, with at least 24 months of data for all patients. RESULTS: There were 73 shoulders that met our inclusion criteria, but 4 underwent revision before 2 years' follow-up. Only 1 of these 4 was revised for aseptic humeral loosening. Sixty-nine shoulders had at least 24 months of radiographic follow-up, and 62 had radiographic and clinical follow-up. Of the 69 shoulders, 5 underwent revision for humeral loosening: 1 for aseptic loosening and 4 for infection. Two other shoulders with humeral loosening were asymptomatic, and the patients refused revision surgery. The overall revision rate for humeral loosening was 8.2% (6 of 73 shoulders). Radiolucent zones of any size were seen in 71.0%, with 8.7% of these shoulders identified as having humeral stems at risk of future loosening. Significant improvements were made in most of the measured clinical outcomes. CONCLUSIONS: A high percentage of radiolucency was seen around the short-stem press-fit humeral components evaluated in this study at short-term follow-up. The overall rates of loosening and revision for the humeral implant examined in this study are higher than those noted in other recent studies evaluating press-fit stems. The cause of radiolucency and humeral loosening for this implant is not fully understood.

Use of Raman spectroscopy to identify active spermatogenesis and Sertoli-cell-only tubules in mice.

Microdissection testicular sperm extraction (micro-TESE) has become the first line therapy to harvest spermatozoa for men with nonobstructive azoospermia. However, the pitfall is that the selection of seminiferous tubules depends on subjective assessment of the colour and size of tubules, which cannot guarantee successful retrieval of spermatozoa. The aim of this study was to determine whether Raman spectroscopy (RS) could distinguish tubules with spermatogenesis from Sertoli-cell-only (SCO) tubules, and potentially serve as a useful tool to improve sperm retrieval rates. Fourteen male adult mice were divided into two groups: SCO group received a single intraperitoneal injection of busulfan (40 mg per kg body weight), and the control group received a placebo dose of 0.9% saline solution. Mice were sacrificed after 4 weeks, and the testicular tissue was assessed by RS and then confirmed with histopathology. The results indicated that tubules with spermatogenesis had intensified Raman peaks at 748, 1124, 1309, 1446 and 1658 cm-1 compared to SCO tubules, except a decreased peak at 1582 cm-1 . RS was able to distinguish the two groups with a sensitivity of 91.2% and specificity of 82.9%. In conclusion, RS may serve as a useful diagnostic tool prior to sperm retrieval.

Prospective study of cyclophosphamide, thiotepa, carboplatin combined with adoptive DC-CIK followed by metronomic cyclophosphamide therapy as salvage treatment for triple negative metastatic breast cancers patients (aged <45)

Background. The recent immunotherapy treatment on triple-negative breast cancer (TNBC) leads to the breakthrough assignation. In this study, we have tried the new combinations of specific chemo with DC-CIKs immunotherapy to treat those patients. Patients and methods. Twenty-three metastatic anthracyclines and taxanes pretreated TNBC younger (mean 41.5 years) patients were initially mobilized with cyclophosphamide (3 g/m2) for the preparation of CD34+ peripheral blood mononuclear cells as the resources for generating DC/CIKs and marrow function supports. All cases were subsequently experienced 2 cycles of chemotherapy with cyclophosphamide 3 g/m2, thiotepa 150 mg/m2, and carboplatin AUC = 6, Q4w. The patients then received 3 infusions of DC-CIKs at the chemo intervals and followed by maintenance therapy with oral cyclophosphamide 50 mg daily. The endpoints were progression-free survival and overall survival. Results. The partial response rate was 13.0 %, stable and progressive disease rates were 56.5 and 30.4 %, respectively. The median PFS was 13.5 months (95 % confidence interval (CI) 10.1-16.9 months) and OS was 15.2 months (95 % CI 12.5-18.1 months). The most common serious adverse events were neutropenia (100.0 %) and anemia (69.7 %) but without treatment-related mortality. Conclusion. These data suggested that such combination therapy model be effective and safe for younger metastatic TNBC exposure to previous anthracyclines and taxanes based adjuvant chemotherapy (AU)

No disponible

Radiographic and clinical outcomes of total shoulder arthroplasty with an all-polyethylene pegged bone ingrowth glenoid component: prospective short- to medium-term follow-up.

BACKGROUND: Glenoid components often cause total shoulder arthroplasty failure. This study examines short-term to midterm radiographic and clinical results of a hybrid glenoid component with 3 cemented peripheral pegs and a central peg, which allows biologic fixation with use of native humeral head autograft. METHODS: In 4 years, 80 glenoid components were implanted during primary total shoulder arthroplasty with at least 2-year follow-up data. Within 12 months, 4 shoulders were revised and excluded from final analyses. Seven patients did not complete their questionnaires. Outcomes data included the American Shoulder and Elbow Surgeons (ASES) questionnaire, Constant score, and satisfaction score. A shoulder and elbow fellowship-trained surgeon, not involved in the care of these patients, analyzed radiographs for radiolucent lines, glenoid seating, and radiodensity in between the flanges of the central peg. RESULTS: Only 1 of 80 shoulders was revised for aseptic glenoid loosening. At final follow-up, 81.6% had a radiolucency grade of 0 or 1. Nearly 90% had a glenoid seating grade of A or B. Grade 2 or 3 bone around the central peg was seen in 88.2%. No statistical association existed between Walch glenoid types and radiolucency grades, bone grades around the central peg, perfect radiolucency grade, seating grade, and grade 3 bone around the central peg. There was significant improvement in mean ASES score, adjusted ASES pain score, Constant score, and satisfaction score as well as in forward flexion, abduction, and external rotation. CONCLUSIONS: The hybrid glenoid can produce stable radiographic and clinical outcomes at short- to medium-term follow-up.

Prospective study of cyclophosphamide, thiotepa, carboplatin combined with adoptive DC-CIK followed by metronomic cyclophosphamide therapy as salvage treatment for triple negative metastatic breast cancers patients (aged <45).

BACKGROUND: The recent immunotherapy treatment on triple-negative breast cancer (TNBC) leads to the breakthrough assignation. In this study, we have tried the new combinations of specific chemo with DC-CIKs immunotherapy to treat those patients. PATIENTS AND METHODS: Twenty-three metastatic anthracyclines and taxanes pretreated TNBC younger (mean 41.5 years) patients were initially mobilized with cyclophosphamide (3 g/m(2)) for the preparation of CD34(+) peripheral blood mononuclear cells as the resources for generating DC/CIKs and marrow function supports. All cases were subsequently experienced 2 cycles of chemotherapy with cyclophosphamide 3 g/m(2), thiotepa 150 mg/m(2), and carboplatin AUC = 6, Q4w. The patients then received 3 infusions of DC-CIKs at the chemo intervals and followed by maintenance therapy with oral cyclophosphamide 50 mg daily. The endpoints were progression-free survival and overall survival. RESULTS: The partial response rate was 13.0 %, stable and progressive disease rates were 56.5 and 30.4 %, respectively. The median PFS was 13.5 months (95 % confidence interval (CI) 10.1-16.9 months) and OS was 15.2 months (95 % CI 12.5-18.1 months). The most common serious adverse events were neutropenia (100.0 %) and anemia (69.7 %) but without treatment-related mortality. CONCLUSION: These data suggested that such combination therapy model be effective and safe for younger metastatic TNBC exposure to previous anthracyclines and taxanes based adjuvant chemotherapy.

CtBP maintains cancer cell growth and metabolic homeostasis via regulating SIRT4.

Cancer cells rely on glycolysis to maintain high levels of anabolism. However, the metabolism of glucose via glycolysis in cancer cells is frequently incomplete and results in the accumulation of acidic metabolites such as pyruvate and lactate. Thus, the cells have to develop strategies to alleviate the intracellular acidification and maintain the pH stability. We report here that glutamine consumption by cancer cells has an important role in releasing the acidification pressure associated with cancer cell growth. We found that the ammonia produced during glutaminolysis, a dominant glutamine metabolism pathway, is critical to resist the cytoplasmic acidification brought by the incomplete glycolysis. In addition, C-terminal-binding protein (CtBP) was found to have an essential role in promoting glutaminolysis by directly repressing the expression of SIRT4, a repressor of glutaminolysis by enzymatically modifying glutamate dehydrogenase in mitochondria, in cancer cells. The loss of CtBP in cancer cells resulted in the increased apoptosis due to intracellular acidification and the ablation of cancer cell metabolic homeostasis represented by decreased glutamine consumption, oxidative phosphorylation and ATP synthesis. Importantly, the immunohistochemistry staining showed that there was excessive expression of CtBP in tumor samples from breast cancer patients compared with surrounding non-tumor tissues, whereas SIRT4 expression in tumor tissues was abolished compared with the non-tumor tissues, suggesting CtBP-repressed SIRT4 expression contributes to the tumor growth. Therefore, our data suggest that the synergistically metabolism of glucose and glutamine in cancer cells contributes to both pH homeostasis and cell growth. At last, application of CtBP inhibitor induced the acidification and apoptosis of breast cancer cells and inhibited glutaminolysis in engrafted tumors, suggesting that CtBP can be potential therapeutic target of cancer treatment.

Usnic acid inhibits ER stress activation through AMPK signaling pathway in rat cardiomyocytes.

OBJECTIVE: In the present study, we investigated the effects of usnic acid (UA) on the endoplasmic reticulum (ER) stress processes in rat cardiomyocytes. MATERIALS AND METHODS: Gene expression of pro-inflammatory cytokines and activation of ER stress signaling were analyzed. Besides, levels of phosphorylated AMPK were measured to evaluate the mechanisms of UA. Finally, small interfering RNA (siRNA) oligos targeting AMPK subunits were used to determine the roles of AMPK in rat cardiomyocytes treated with UA. RESULTS: We found that UA treatment significantly reduced ER stress activation and expression of pro-inflammatory cytokines, via an AMPK signaling-dependent manner. CONCLUSIONS: UA might be useful to reduce the occurrence of adverse cardiovascular events.

Higher-order structure of the Rous sarcoma virus SP assembly domain.

UNLABELLED: Purified retroviral Gag proteins can assemble in vitro to form immature virus-like particles (VLPs). By cryoelectron tomography, Rous sarcoma virus VLPs show an organized hexameric lattice consisting chiefly of the capsid (CA) domain, with periodic stalk-like densities below the lattice. We hypothesize that the structure represented by these densities is formed by amino acid residues immediately downstream of the folded CA, namely, the short spacer peptide SP, along with a dozen flanking residues. These 24 residues comprise the SP assembly (SPA) domain, and we propose that neighboring SPA units in a Gag hexamer coalesce to form a six-helix bundle. Using in vitro assembly, alanine scanning mutagenesis, and biophysical analyses, we have further characterized the structure and function of SPA. Most of the amino acid residues in SPA could not be mutated individually without abrogating assembly, with the exception of a few residues near the N and C termini, as well as three hydrophilic residues within SPA. We interpret these results to mean that the amino acids that do not tolerate mutations contribute to higher-order structures in VLPs. Hydrogen-deuterium exchange analyses of unassembled Gag compared that of assembled VLPs showed strong protection at the SPA region, consistent with a higher-order structure. Circular dichroism revealed that a 29mer SPA peptide shifts from a random coil to a helix in a concentration-dependent manner. Analytical ultracentrifugation showed concentration-dependent self-association of the peptide into a hexamer. Taken together, these results provide strong evidence for the formation of a critical six-helix bundle in Gag assembly. IMPORTANCE: The structure of a retrovirus like HIV is created by several thousand molecules of the viral Gag protein, which assemble to form the known hexagonal protein lattice in the virus particle. How the Gag proteins pack together in the lattice is incompletely understood. A short segment of Gag known to be critical for proper assembly has been hypothesized to form a six-helix bundle, which may be the nucleating event that leads to lattice formation. The experiments reported here, using the avian Rous sarcoma virus as a model system, further define the nature of this segment of Gag, show that it is in a higher-order structure in the virus particle, and provide the first direct evidence that it forms a six-helix bundle in retrovirus assembly. Such knowledge may provide underpinnings for the development of antiretroviral drugs that interfere with virus assembly.

In Vitro Assembly of Retroviruses.

Assembly, part of the late stages of the retroviral life cycle, begins when the structural polyprotein Gag associates with viral genomic RNA. Ultimately, more than a thousand Gag molecules form a spherical immature virion. Maturation takes place soon after or concomitantly with virus budding and is initiated as Gag is cleaved by the retroviral protease into its constituent protein domains. The immature core is thought to disassemble and the liberated CA proteins to reassemble into a morphologically distinct mature capsid. In vitro assembly with derivatives of Gag and CA has been used to study retroviruses for over two decades. In this review, we examine the discovery and development of three major model systems [human immunodeficiency virus type 1 (HIV-1), Rous sarcoma virus (RSV), and Mason-Pfizer monkey virus (MPMV)] and discuss structural features and aspects of the retroviral assembly pathway that have been uncovered using in vitro assembly. We also put forward two major unresolved questions in the field and propose future avenues of research.

Plasma chemerin level in metabolic syndrome.

To investigate the chemerin level in the Chinese Han population with metabolic syndrome and its relationship with each metabolic syndrome component [body mass index (BMI), blood pressure, blood lipids, and blood glucose], we selected 30 patients with metabolic syndrome and 30 healthy control subjects. The chemerin level was measured by enzyme immunoassay in these 2 groups. The subjects' weight, blood pressure, BMI, waist circumference, fasting blood glucose, fasting insulin, lipids, and glycated hemoglobin were simultaneously detected. The t-test, correlation analysis, and multiple regression analysis were used to perform statistical analysis. We found that plasma chemerin level was higher in the metabolic syndrome group than that in the control group (97.61 ± 6.49 vs 70.26 ± 6.97, t = 15.73, P < 0.05). The plasma chemerin level was positively correlated with systolic blood pressure, waist circumference, BMI, waist-to-hip ratio, fasting blood glucose, fasting insulin, and glycated hemoglobin (r = 0.548, 0.442, 0.359, 0.556, 0.613, 0.581, and 0.572, respectively; all P < 0.05). However, it was negatively correlated with high-density lipoprotein cholesterol (r = -0.378, P < 0.05). Therefore, we concluded that plasma chemerin level was correlated with obesity, blood pressure, and high-density lipoprotein cholesterol, suggesting that it may play a role in the pathogenesis of metabolic syndrome.

Selections of appropriate regimen of high-dose chemotherapy combined with adoptive cellular therapy with dendritic and cytokine-induced killer cells improved progression-free and overall survival in patients with metastatic breast cancer: reargument of such contentious therapeutic preferences

BACKGROUND: We hypothesized that combination of dendritic cell (DC) with autologous cytokine-induced killer (CIK) immunotherapy in setting of high-dose chemotherapy (HDC) would be effective for selected metastatic breast cancer (MBC) patients. PATIENTS AND METHODS: Our previous work showed thiotepa could eradicate breast cancer stem cells. From 2004 to 2009, 79 patients received standard dose chemotherapy (SDC) of 75 mg/m(2) docetaxel and 75 mg/m(2) thiotepa versus 87 patients of HDC + DC/CIK: 120 mg/m(2) docetaxel to mobilize peripheral CD34(+) progenitor cells, a sequence of HDC (120 mg/m(2) docetaxel, plus 175 mg/m(2) thiotepa) + DC/CIK, with or without 400 mg/m(2) carboplatin depending upon bone marrow function. The endpoints were response rates (RR), progression-free survival (PFS), and overall survival (OS). RESULTS: Compared with SDC, PFS and OS were improved in HDC + DC/CIK (median PFS 10.2 vs. 3.7 months, P < 0.001; median OS 33.1 vs. 15.2 months, P < 0.001). Patients of pre-menopausal, HDC as first-line treatment after metastasis, or with visceral metastasis showed prolonged PFS and OS. SDC group also achieved the similar response as previous reports. CONCLUSION: Our study demonstrated the novel combination of HDC with DC/CIK to be an effective choice for the selected MBC population, in which choosing appropriate chemo regimens played important roles, and also specific HDC regimen plus DC/CIK immunotherapy showed the clinical benefits compared with chemotherapy alone (AU)

Specific genetic polymorphisms of IL 10-592 AA and IL 10-819 TT genotypes lead to the key role for inducing docetaxel-induced liver injury in breast cancer patients

AIM: This study was designed to explore the genetic polymorphism of IL-10 (-1082A/G, -592A/C, -819T/C), TNF-α (-308G/A) with susceptibility to docetaxel-induced liver injury (DILI) in Chinese breast cancer patients. METHODS: The targeted genetic polymorphisms of IL10-1082G/A, IL10-592A/C, IL10-819T/C, TNF-308G/A from 40 patients with DILI were assayed by matrix-assisted laser desorption/ionization-time of flight of Sequenom. RESULTS: AA genotype of IL10-592 and TT of IL10-819 significantly increased incidence of DILI (P = 0.005, OR = 3.137). No differences of TNF gene polymorphism between the two groups were seen. CONCLUSION: The genetic polymorphism of the IL10-592A/C AA genotype and IL10-819T/C TT genotype was predominantly conferred to the incidence of docetaxel-induced liver injury (AU)