Impact of cytotoxic T lymphocytes immunotherapy on prognosis of colorectal cancer patients.

Background: Expansion and activation of cytotoxic T lymphocytes (CTLs) in vitro represents a promising immunotherapeutic strategy, and CTLs can be primed by dendritic cells (DCs) loaded with tumor-associated antigens (TAAs) transformed by recombinant adeno-associated virus (rAAV). This study aimed to explore the impact of rAAV-DC-induced CTLs on prognosis of CRC and to explore factors associated with prognosis. Methods: This prospective observational study included patients operated for CRC at Yan'an Hospital Affiliated to Kunming Medical University between 2016 and 2019. The primary outcome was progression-free survival (PFS), secondary outcomes were overall survival (OS) and adverse events. Totally 49 cases were included, with 29 and 20 administered rAAV-DC-induced CTL and chemotherapy, respectively. Results: After 37-69 months of follow-up (median, 54 months), OS (P=0.0596) and PFS (P=0.0788) were comparable between two groups. Mild fever occurred in 2 (6.9%) patients administered CTL infusion. All the chemotherapy group experienced mild-to-moderate adverse effects, including vasculitis (n=20, 100%), vomiting (n=5, 25%), nausea (n=17, 85%) and fatigue (n=17, 85%). Conclusions: Lymphatic metastasis (hazard ratio [HR]=4.498, 95% confidence interval [CI]: 1.290-15.676; P=0.018) and lower HLA-I expression (HR=0.294, 95%CI: 0.089-0.965; P=0.044) were associated with poor OS in the CTL group. CTLs induced by rAAV-DCs might achieve comparable effectiveness in CRC patients compare to chemotherapy, cases with high tumor-associated HLA-I expression and no lymphatic metastasis were more likely to benefit from CTLs.

Elevated CTSL Gene Expression Correlated with Proinflammatory Cytokines in Omental Adipose Tissue of Patients with Obesity.

Purpose: Cathepsin L (CTSL) and B (CTSB) were lysosomal proteases, and their expression and activity contribute to the progression of inflammation in obese rodents. Our aim was to investigate CTSB and CTSL expression in omental adipose tissue (AT) of patients with obesity and to correlate CTSB and CTSL expression with proinflammatory cytokines (CCL-2, IL-6 and IL-1ß). Patients and Methods: A total of 12 patients without obesity (NOB) and 51 patients with obesity (OB) were involved in this study. Omental AT was collected from all the participants for RNA extraction. Expressions of CTSB, CTSL and proinflammatory cytokines (CCL-2, IL-6 and IL-1ß) were qualified with qRT-PCR. BMI (body mass index) and metabolic parameters were measured. Results: The mRNA expression levels of both CTSB and CTSL were upregulated in the OB group (t = 2.693, P < 0.05; t = 2.849, P<0.01) and were related to TC levels (Std.ß=0.443, P<0.05; Std.ß=0.439, P<0.05). However, only the CTSB level was related to BMI (Std.ß=0.261, P<0.05). In multiple regression analysis, CTSL was independently associated with CCL-2, IL-6 and IL-1ß levels (Std.ß=0.352-0.462, P<0.05). Conclusion: CTSB and CTSL gene expressions were elevated in the omental AT of OB group. CTSL, but not CTSB, was positively correlated with proinflammatory cytokines independently, suggesting that the dysregulation of CTSL may play a significant role in the inflammatory process.

EPHX2 Inhibits Colon Cancer Progression by Promoting Fatty Acid Degradation.

Tumor cells use metabolic reprogramming to keep up with the need for bioenergy, biosynthesis, and oxidation balance needed for rapid tumor division. This phenomenon is considered a marker of tumors, including colon cancer (CRC). As an important pathway of cellular energy metabolism, fatty acid metabolism plays an important role in cellular energy supply and oxidation balance, but presently, our understanding of the exact role of fatty acid metabolism in CRC is limited. Currently, no lipid metabolism therapy is available for the treatment of CRC. The establishment of a lipidmetabolism model regulated by oncogenes/tumor suppressor genes and associated with the clinical characteristics of CRC is necessary to further understand the mechanism of fatty acid metabolism in CRC. In this study, through multi-data combined with bioinformatic analysis and basic experiments, we introduced a tumor suppressor gene, EPHX2, which is rarely reported in CRC, and confirmed that its inhibitory effect on CRC is related to fatty acid degradation.

Repeat hepatic resection versus percutaneous ablation for the treatment of recurrent hepatocellular carcinoma: meta-analysis.

BACKGROUND: The efficacy of repeat hepatic resection (rHR) in the treatment of recurrent hepatocellular carcinoma compared with radiofrequency or microwave ablation after resection of the primary tumour remains controversial. A systematic review and meta-analysis were performed to compare the safety and efficacy of these procedures. METHODS: PubMed, Embase, Scopus, Cochrane Library, and China National Knowledge Infrastructure databases were systematically searched to identify related studies published before 10 October 2021. Overall and recurrence-free survival after different treatments were compared based on pooled hazard ratios with a random-effects model. RESULTS: Two randomized clinical trials and 28 observational studies were included, involving 1961 and 2787 patients who underwent rHR and ablation respectively. Median perioperative mortality in both groups was zero but patients in the rHR group had higher median morbidity rates (17.0 per cent) than those in the ablation group (3.3 per cent). rHR achieved significantly longer recurrence-free survival than ablation (HR 0.79, 95 per cent c.i. 0.70 to 0.89, P < 0.001), while both groups had similar overall survival (HR 0.93, 95 per cent c.i. 0.83 to 1.04, P = 0.18). CONCLUSION: rHR and ablation based on radio- or microwaves are associated with similar overall survival in patients with recurrent hepatocellular carcinoma after resection of the primary tumour.

An Amorphous Peri-Implant Ligament with Combined Osteointegration and Energy-Dissipation.

Progress toward developing metal implants as permanent hard-tissue substitutes requires both osteointegration to achieve load-bearing support, and energy-dissipation to prevent overload-induced bone resorption. However, in existing implants these two properties can only be achieved separately. Optimized by natural evolution, tooth-periodontal-ligaments with fiber-bundle structures can efficiently orchestrate load-bearing and energy dissipation, which make tooth-bone complexes survive extremely high occlusion loads (>300 N) for prolonged lifetimes. Here, a bioinspired peri-implant ligament with simultaneously enhanced osteointegration and energy-dissipation is presented, which is based on the periodontium-mimetic architecture of a polymer-infiltrated, amorphous, titania nanotube array. The artificial ligament not only provides exceptional osteoinductivity owing to its nanotopography and beneficial ingredients, but also produces periodontium-similar energy dissipation due to the complexity of the force transmission modes and interface sliding. The ligament increases bone-implant contact by more than 18% and simultaneously reduces the effective stress transfer from implant to peri-implant bone by ≈30% as compared to titanium implants, which as far as is known has not previously been achieved. It is anticipated that the concept of an artificial ligament will open new possibilities for developing high-performance implanted materials with increased lifespans.

Decreased m6A Modification of CD34/CD276(B7-H3) Leads to Immune Escape in Colon Cancer.

Previous studies have reported that m6a modification promotes tumor immune escape by affecting tumor microenvironment (TME). Due to the complexity of TME, a single biomarker is insufficient to describe the complex biological characteristics of tumor and its microenvironment. Therefore, it is more meaningful to explore a group of effective biomarkers reflecting different characteristics of cancer to evaluate the biological characteristics of solid tumors. Here, the immune gene CD34/CD276 with different m6A peak was obtained by m6A sequencing (MeRIP-seq) of colon cancer (CRC)clinical samples and combined with MsIgDB database, which was used to perform cluster analysis on TCGA-COAD level 3 data. The CD34/CD276 as a molecular marker for CRC prognosis was confirmed by survival analysis and immunohistochemical assay. Further bioinformatics analysis was carried out to analyze the molecular mechanism of CD34/CD276 affecting the TME through m6a-dependent down-regulation and ultimately promoting immune escape of CRC.

Pyrazinamide Resistance and Mutation Patterns Among Multidrug-Resistant Mycobacterium tuberculosis from Henan Province.

PURPOSE: This study was designed to identify the phenotypic and genotypic characteristics of pyrazinamide (PZA) resistance among multidrug-resistant Mycobacterium tuberculosis (MDR-TB) from Henan and to evaluate the efficacy of pncA, rpsA, and panD mutations in predicting PZA resistance. MATERIALS AND METHODS: A total of 152 MDR strains were included in this study. The Bactec MGIT system was used to determine PZA susceptibility for all strains. The pncA, rpsA, and panD genes were sequenced to identify any mutations, and the sequences were then aligned with the sequence of standard strain H37Rv. Moreover, the correlations between PZA-resistant phenotypes and treatment outcomes were analysed. RESULTS: Of the152 strains, 105 had a PZA-resistant phenotype, and 102 harboured the pncA mutation. The PZA resistance rate was higher in the strains with resistance to all four first-line drugs and those that were pre-extensively drug-resistant (pre-XDR) and extensively drug-resistant (XDR). A total of 100 different pncA mutation patterns were identified, including 80 point mutations and 20 insertions/deletions, and 32 new pncA mutation patterns were detected. In this study, 13 strains had multiple mutations. Of the11 PZA-resistant strains without pncA mutations, two harboured the rpsA mutation, and one harboured the panD mutation. With PZA susceptibility results as the reference, single-gene pncA sequencing had sensitivity of 89.52% and specificity of 89.36%. With the combination of rpsA and panD, the sensitivity increased to 92.38%, and the specificity remained the same. No significant differences were observed in the sputum smear/culture conversion rate between PZA-resistant patients and PZA-sensitive patients. However, PZA resistance was related to the time to sputum smear/culture conversion (P = 0.018). CONCLUSION: The combination of pncA, rpsA, and panD was beneficial for the timely diagnosis of PZA resistance and could provide a laboratory basis for customizing treatment regimens for MDR-TB patients.

Adaptive structural reorientation: Developing extraordinary mechanical properties by constrained flexibility in natural materials.

Seeking strategies to enhance the overall combinations of mechanical properties is of great significance for engineering materials, but still remains a key challenge because many of these properties are often mutually exclusive. Here we reveal from the perspective of materials science and mechanics that adaptive structural reorientation during deformation, which is an operating mechanism in a wide variety of composite biological materials, functions more than being a form of passive response to allow for flexibility, but offers an effective means to simultaneously enhance rigidity, robustness, mechanical stability and damage tolerance. As such, the conflicts between different mechanical properties can be "defeated" in these composites merely by adjusting their structural orientation. The constitutive relationships are established based on the theoretical analysis to clarify the effects of structural orientation and reorientation on mechanical properties, with some of the findings validated and visualized by computational simulations. Our study is intended to give insight into the ingenious designs in natural materials that underlie their exceptional mechanical efficiency, which may provide inspiration for the development of new man-made materials with enhanced mechanical performance. STATEMENT OF SIGNIFICANCE: It is challenging to attain certain combinations of mechanical properties in man-made materials because many of these properties - for example, strength with toughness and stability with flexibility - are often mutually exclusive. Here we describe an effective solution utilized by natural materials, including wood, bone, fish scales and insect cuticle, to "defeat" such conflicts and elucidate the underlying mechanisms from the perspective of materials science and mechanics. We show that, by adaptation of their structural orientation on loading, composite biological materials are capable of developing enhanced rigidity, strength, mechanical stability and damage tolerance from constrained flexibility during deformation - combinations of attributes that are generally unobtainable in man-made systems. The design principles extracted from these biological materials present an unusual yet potent new approach to guide the development of new synthetic composites with enhanced combinations of mechanical properties.

Multiscale designs of the chitinous nanocomposite of beetle horn towards an enhanced biomechanical functionality.

Operating mainly as a type of weapon, the beetle horn develops an impressive mechanical efficiency based on chitinous materials to maximize the injury to opponent and simultaneously minimize the damage to itself and underlying brain under stringent loading conditions. Here the cephalic horn of the beetle Allomyrina dichotoma is probed using multiscale characterization combined with finite element simulations to explore the origins of its biomechanical functionality from the perspective of materials science. The horn is revealed to be highly regulated from the macroscopic shape, geometry, and connection with the body to the meso- and microscopic architecture, moisture content, and chemical and structural characteristics. Varying kinds of gradients are integrated at all length-scales. Such designs are demonstrated to benefit the mechanical performance by mitigating stress concentrations, retarding crack propagation, and modulating local properties to better adapt to stress. Enhanced rigidity, robustness and stability are additionally generated from the constrained flexibility endowed by the nanocomposite plywood structure through the reorientation of chitin nanofibrils within the proteinaceous matrix. These findings shed light on the intriguing materials-design strategies of nature in creating synergy of offence and persistence. They may even offer inspiration for the synthesis of high-performance materials and structures, in particular beams to resist bending and torsion.

Study on the metabolites of DH-e, a Halomonas marine bacterium, against three toxic dinoflagellate species.

Algicidal bacteria play an important role in mitigating harmful algal blooms (HABs). In the study, five bacterial strains were isolated from the East China Sea. One strain of algicidal bacterium, named DH-e, was found to selectively inhibit the motor ability of Prorocentrum donghaiense, Alexandrium tamarense (ATDH-47) and Karenia mikimotoi Hansen. Both 16S rDNA sequence analysis and morphological characteristics revealed that the algicidal DH-e bacterium belonged to Halomonas. Furthermore, results showed that the metabolites in the DH-e cell-free filtrate could kill algae directly, and the minimum inhibitory concentrations (MICs) of the bacterial metabolites on the cells of the three dinoflagellate species ranged from 35.0-70.0 µg/mL. Following short-term inhibitory tests, the dinoflagellates in mixed crude extract solution (0.7 mg/mL) ceased movement after 5 min. The algicidal mechanism of the metabolites was investigated through enzyme activities, including that of catalase (CAT), alkaline phosphatase (AKP), acetone peroxide (T-ATP) synthetase and nitrite reductase (NR). Results indicated that metabolites did not disrupt the energy or nutrient routes of the algae (P > 0.05), but did initiate an increase in free radicals in the algal cells, which might explain the subsequent death of sensitive algae. Thus, the metabolites of the DH-e bacterium showed promising potential for controlling HABs.

Role of MIRU-VNTR and spoligotyping in assessing the genetic diversity of Mycobacterium tuberculosis in Henan Province, China.

BACKGROUND: Tuberculosis remains a serious threat to human health as an infectious disease in China. Henan, a most populated province in China, has a high incidence of tuberculosis (TB). Though the genetic diversity of Mycobacterium tuberculosis (MTB) has been investigated in many regions, there have been only a few studies on the molecular characteristics and drug resistance phenotypes in Henan. This is the first study on the genetic profile of MTB from Henan. METHODS: A total of 668 MTB isolates from various areas were genotyped with spoligotyping and 26-locus MIRU-VNTR (classical 24-locus MIRU-VNTR and 2 other loci). The association between TB spoligotype signatures and drug-resistant profiles was analysed. RESULTS: Our data revealed that MTB isolates circulating in Henan had a high degree of genetic variation. The Beijing family was the most predominant genotype (83.53%,n = 558), and the typical Beijing type(ST1) was the major sublineage (81.73%,n = 546). In total,668 isolates were divided into 567 different types, forming 38 clusters (2-15 isolates per cluster), and 529 unique types by 26-locus MIRU-VNTR analysis. There was no correlation between the Beijing family and gender, age at diagnosis or treatment history, whereas the Beijing family was significantly associated with all four first-line drug resistance and multidrug-resistant phenotypes. For these samples, 15 of 26 MIRU-VNTR loci had high or moderate discriminatory power according to the Hunter-Gaston discriminatory index. A combination of the 10 most polymorphic loci had similar discriminatory power as the 26-locus set. CONCLUSION: The Beijing genotype is the most prevalent family. Ten-locus MIRU-VNTR in combination with spoligotyping can efficiently classify the molecular type of MTB in Henan Province.

MiR-330-3p inhibits gastric cancer progression through targeting MSI1.

Increasing evidences demonstrated that microRNAs (miRNAs) play critical roles in the human tumor development and progression. In our study, we found that miR-330-3p expression was downregulated in gastric cancer cell lines and tissues. Ectopic expression of miR-330-3p suppressed the gastric cancer cell proliferation, colony formation and migration. Overexpression of miR-330-3p promoted E-cadherin expression and inhibited the expression of N-cadherin, vimentin and snail. We identified Musashi-1 (MSI1) as a direct target gene of miR-330-3p in gastric cancer cell. In addition, MSI1 was upregulated in gastric cancer cell lines and tissues and the MSI1 expression was inversely correlated with miR-330-3p expression in gastric cancer tissues. MiR-330-3p expression was increased in gastric cancer cells after treated with histone deacetylase inhibitor trichostatin A (TSA) and DNA methylation inhibitor 5-aza-CdR (AZA). These indicated that downregulated expression of miR-330-3p was partly mediated by gene promoter region hypermethylation. These results suggested that miR-330-3p acted as a tumor suppressor gene in GC.

Enhanced protective role in materials with gradient structural orientations: Lessons from Nature.

UNLABELLED: Living organisms are adept at resisting contact deformation and damage by assembling protective surfaces with spatially varied mechanical properties, i.e., by creating functionally graded materials. Such gradients, together with multiple length-scale hierarchical structures, represent the two prime characteristics of many biological materials to be translated into engineering design. Here, we examine one design motif from a variety of biological tissues and materials where site-specific mechanical properties are generated for enhanced protection by adopting gradients in structural orientation over multiple length-scales, without manipulation of composition or microstructural dimension. Quantitative correlations are established between the structural orientations and local mechanical properties, such as stiffness, strength and fracture resistance; based on such gradients, the underlying mechanisms for the enhanced protective role of these materials are clarified. Theoretical analysis is presented and corroborated through numerical simulations of the indentation behavior of composites with distinct orientations. The design strategy of such bioinspired gradients is outlined in terms of the geometry of constituents. This study may offer a feasible approach towards generating functionally graded mechanical properties in synthetic materials for improved contact damage resistance. STATEMENT OF SIGNIFICANCE: Living organisms are adept at resisting contact damage by assembling protective surfaces with spatially varied mechanical properties, i.e., by creating functionally-graded materials. Such gradients, together with multiple length-scale hierarchical structures, represent the prime characteristics of many biological materials. Here, we examine one design motif from a variety of biological tissues where site-specific mechanical properties are generated for enhanced protection by adopting gradients in structural orientation at multiple length-scales, without changes in composition or microstructural dimension. The design strategy of such bioinspired gradients is outlined in terms of the geometry of constituents. This study may offer a feasible approach towards generating functionally-graded mechanical properties in synthetic materials for improved damage resistance.

The feasibility of laparoscopic splenectomy for ITP patients without preoperative platelet transfusion.

BACKGROUND/AIMS: The aim of this study was to investigate the feasibility and safety of laparoscopic splenectomy for idiopathic thrombocytopenic purpura patients without platelet transfusion before surgery. METHODOLOGY: From 2004 to 2010, laparoscopic splenectomy was performed in 30 patients with ITP (idiopathic thrombocytopenic purpura, ITP) in our department and clinical information was recorded. Inclusion criteria is patient with a preoperative platelet count of <20x109/L. All of the 30 patients retrospectively reviewed were divided into 2 groups, according to whether they received platelet transfusion preoperatively or not. Perioperative data were analyzed. RESULTS: The clinical efficacy of treatment was significant (obvious response rates of the two cohorts were 17/19 and 9/10). There was no significant difference in the surgical procedure or postoperative efficacy of laparoscopic splenectomy for ITP patients, regardless of whether there was preoperative platelet transfusion, including operating time, ratio of intraoperative blood transfusion, estimated blood loss, the rate of conversion, postoperative platelet counts, amount of postoperative drainage and postoperative hospital stay. CONCLUSIONS: Laparoscopic splenectomy without platelet transfusion can be a feasible and safe procedure in patients with a platelet count lower than 20x109/L, with a normal prothrombin time and without any preoperative bleeding tendency or severe coagulopathy.

Skewed T-cell subsets and enhanced macrophages phagocytosis in the spleen of patients with immune thrombocytopenia failing glucocorticoids.

T-cell abnormalities are considered important in the pathogenesis of immune thrombocytopenia (ITP). Th1 polarization of the immune response in peripheral blood mononuclear cells (PBMC) has been observed in ITP patients. However, whether the polarization of T-cell subsets exists in splenocytes remains unclear. In the present study, we detected T-cell subsets and macrophage phagocytosis capacity in the spleens of ten ITP patients failing first-line treatment. Compared with traumatic patients, ITP patients had similar percentages of Th1, Tc1, and T1 (Th1 + Tc1) cells, but lower percentages of Th2 and Tc2 cells, which did not, however, reach significant difference. ITP patients had significantly lower percentages of T2 (Th2 + Tc2) cells than the control group (P = 0.035). The ratios of Th1/Th2 and T1/T2 were significantly higher in ITP patients than the control group (P = 0.025 and 0.027, respectively). Splenic macrophages from ITP patients presented significant increase in phagocytic capacity (P < 0.001). The phagocytosis rate in splenic macrophages positively correlated with the ratio of Th1/Th2 (r = 0.767, P = 0.010), but negatively correlated with percentage of Th2 cells (r = -0.804, P = 0.005). The skewed T-cell subsets and enhanced macrophage phagocytosis in the spleen may be closely related to the disease onset and status of ITP.