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Zn 2+ -dependent histone deacetylases (HDACs) are enzymes that regulate gene expression by removing acetyl groups from histone proteins. These enzymes are essential in all living systems, playing key roles in cancer treatment and as potential pesticide targets. Previous phylogenetic analyses of HDAC in certain species have been published. However, their classification and evolutionary origins across biological kingdoms remain unclear, which limits our understanding of them. In this study, we collected the HDAC sequences from 1451 organisms and performed analyses. The HDACs are found to diverge into three classes and seven subclasses under divergent selection pressure. Most subclasses show species specificity, indicating that HDACs have evolved with high plasticity and diversification to adapt to different environmental conditions in different species. In contrast, HDAC1 and HDAC3, belonging to the oldest class, are conserved and crucial in major kingdoms of life, especially HDAC1. These findings lay the groundwork for the future application of HDACs.

Although electrospinning (e-spinning) has witnessed rapid development in recent years, it has also been criticized by environmentalists due to the use of organic solvents. Therefore, aqueous e-spinning (green e-spinning) is considered a more attractive technique. However, considering the poor water resistance and mechanical properties of electrospun (e-spun) nanofibers, cross-linking is a perfect solution. In this review, we systematically discuss the cross-linking e-spinning system for the first time, including cross-linking strategies ( in situ , liquid immersion, vapor, and spray cross-linking), cross-linking mechanism (physical and chemical cross-linking) of e-spun nanofibers, and the various applications ( e.g. , tissue engineering, drug delivery, water treatment, food packaging, and sensors) of cross-linked e-spun nanofibers. Among them, we highlight several cross-linking methods, including UV light cross-linking, electron beam cross-linking, glutaraldehyde (and other commonly used cross-linking agents) chemical cross-linking, thermal cross-linking, and enzymatic cross-linking. Finally, we confirm the significance of cross-linking e-spinning and reveal the problems in the construction of this system.

Radiation encephalopathy (RE) is an important potential complication in patients with nasopharyngeal carcinoma (NPC) who undergo radiotherapy (RT) that can affect the quality of life. However, a functional imaging biomarker of pre-symptomatic RE has not yet been established. This study aimed to assess radiation-induced gray matter functional alterations and explore fractional amplitude of low-frequency fluctuation (fALFF) as an imaging biomarker for predicting or diagnosing RE in patients with NPC. A total of 60 patients with NPC were examined, 21 in the pre-RT cohort and 39 in the post-RT cohort. Patients in the post-RT cohort were further divided into two subgroups according to the occurrence of RE in follow-up: post-RT non-RE ( n = 21) and post-RT REproved infollow-up ( n = 18). Surface-based and volume-based fALFF were used to detect radiation-induced functional alterations. Functional derived features were then adopted to construct a predictive model for the diagnosis of RE. We observed that surface-based fALFF could sensitively detect radiation-induced functional alterations in the intratemporal brain regions (such as the hippocampus and superior temporal gyrus), as well as the extratemporal regions (such as the insula and prefrontal lobe); however, no significant intergroup differences were observed using volume-based fALFF. No significant correlation between fALFF and radiation dose to the ipsilateral temporal lobe was observed. Support vector machine (SVM) analysis revealed that surface-based fALFF in the bilateral superior temporal gyri and left insula exhibited impressive performance (accuracy = 80.49%) in identifying patients likely to develop RE. We conclude that surface-based fALFF may serve as a sensitive imaging biomarker in the prediction of RE., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhang, Kang, Zeng, Li, Gao, Liu, Shi and Liao.)

Objective To investigate the expressions,roles,and clinical significance of microRNA-365(miR-365)and E74-like factor 4(ELF4)in cervical cancer. Methods The expressions of miR-365 in normal cervical tissues(n=34),cervical intraepithelial neoplasia 1(CIN 1)(n=31),cervical intraepithelial neoplasia2-3(CIN 2-3)(n=37),squamous cell carcinoma of the cervix(SCC)(n=33),and three cervical cancer cell lines(C33A cells,Hela cells,and SiHa cells)were detected by real-time quantitative polymerase chain reaction(qPCR).Bioinformatic prediction and luciferase reporter gene assay were performed to verify whether ELF4 was a direct target of miR-365.Western blot and immunohistochemistry were used to detect ELF4 expression in cervical cancer cells and in different pathological cervix tissues.CCK8 assay was used to detect the effect of overexpression or inhibition of miR-365 on the proliferation of cervical cancer cells at different time points.The relationships among the miR-365 expression,ELF4 expression,and clinicopathological parameters of cervical cancer were analyzed by correlation analysis. Results qPCR results showed that compared with the normal cervical cell HcerEpic,the expressions of miR-365 in CIN1,CIN2-3,and cervical cancer tissues gradually decreased with the increased pathologic grade,and its expressions also decreased in different cervical cancer cell lines.The luciferase reporter gene assay confirmed that ELF4 was the direct target of miR-365.Western blot showed that the expression of ELF4 increased in all three cervical cancer cell lines compared with normal cervical epidermal cell(P=0.013,P=0.002,P=0.004).Immunohistochemistry showed that ELF4 expression was up-regulated in CIN and cervical cancer tissues.CCK8 assay showed that overexpression of miR-365 inhibited cell proliferation,while inhibition of miR-365 promoted the proliferation of three cervical cancer cells(P<0.05).Further analysis confirmed that there was a negative correlation between the expression levels of miR-365 and ELF4 in CIN2-3 and SCC(r=-0.351,P=0.045;r=-0.349,P=0.035).Clinical analysis showed that the expressions of both miR-365 and ELF4 were correlated with tumor size,pathological grade,and clinical stage in SCC(all P < 0.05). Conclusion The decreased expression of miR-365 in human cervical cancer cells relieves its inhibitory effect on ELF4,which promotes the proliferation of cervical cancer cells and the formation of tumor.

Backgrounds: The present study aimed to evaluate benefit of hepatic arterial infusion chemotherapy (HAI) combined with systemic chemotherapy (SCT) for patients with colorectal liver metastases (CLMs) in a palliative setting., Methods: This was a retrospective single-center study including 43 consecutive patients with CLM after failure of standard SCT. Among them, 20 (47 %) patients underwent HAI combined with SCT (Group A) and 23 historical control patients who had received SCT with or without targeted agent treatment (Group B)., Results: The two groups had similar characteristics. Compared with SCT alone, HAI combined with SCT prolonged survival (median 19.8 vs. 9.0 months; P = 0.045). Median hepatic progression-free survival was significantly longer for HAI combined with SCT vs. SCT alone (median 8.1 vs. 4.7 months; P = 0.027), as were response rates (25 and 0 %; P = 0.038) and progression-free survival (median 5.7 vs. 3.0 months; P = 0.02). Three patients (15 %) achieved conversion to potentially curative surgery. Grade 3/4 toxicities for Group A and Group B were neutropenia (5 and 8.7 %, respectively), anemia (5 and 0 %, respectively), and hyperbilirubinemia (0 and 4.3 %, respectively). Other complications were mostly grade 1 or 2., Conclusions: HAI combined with SCT treatment can improve overall survival compared with SCT alone in highly advanced CLM refractory to intravenous chemotherapy.

Radiofrequency ablation (RFA) causes coagulative necrosis of tumor tissue and the production of local tumor protein debris. These fragments of tumor protein debris contain a large number of various antigens, which can stimulate a specific cellular immune response. In the present study, dendritic cells (DCs) were loaded with tumor protein lysate antigens that were produced in situ by RFA, and were used to treat murine colon carcinoma in combination with cytokine-induced killer (CIK) cells. Subsequent to the treatment of murine colon carcinoma by RFA, the in situ supernatant of tumor lysis was collected and the DCs were loaded with the lysate antigen to generate Ag-DCs. CIK cells induced from the spleen cells of mice were co-cultured with Ag-DCs to generate Ag-DC-CIK cells. The results revealed that the Ag-DC-CIK cells exhibited strong antitumor activity in vitro and in vivo . The morphology and immunophenotypes of these cells were determined using microscopy and flow cytometry, respectively. The cytotoxic activity of Ag-DC-CIK cells was determined using a CCK-8 assay. To establish a mouse model, mice were randomized into Ag-DC-CIK, DC-CIK, CIK and PBS control groups and monitored for tumor growth and survival time. ANOVA was used to compare the trends in the three groups for implanted tumor volumes. The log-rank test was used to compare the survival time. The present findings indicated that DCs loaded with the protein lysate antigens of tumors, produced in situ by RFA, combined with CIK cells may be a novel strategy for cancer treatment.

A drought-induced gene, DIP3, encoding a chitinase III protein was isolated from the roots of upland rice by reverse transcription-polymerase chain reaction (RT-PCR). Sequence analysis demonstrated that the cDNA and deduced protein showed high identity to Oryza sativa class III chitinase. The deduced protein contained a signal peptide sequence in the N-terminal region of 21aa and a conserved glycosyl hydrolase (GH) 18 domain. The secondary and 3D structures were analyzed and showed that it contained α-helix, β-sheets, extended strand and random coil structures and that it was approximately spheroidal. Real-time quantitative PCR analysis revealed that expression levels accumulated rapidly under different forms of abiotic stress (drought, salt and low temperature), peaked at different times and then decreased. These results implied that as a member of class III chitinases, DIP3 may function as a stress-induced protein involved in the regulation of plant stress response.

The effects of supplementing a barley-based diet for weaned piglets withexogenous beta-glucanase and xylanase on gastrointestinal digestiveenzyme activities were investigated. Thirty-six cross-bred weaned pigletswere randomly assigned to two groups with three pens based on sexand mass. Each group was fed on the diet based on barley with or withoutadded beta-glucanase and xylanase (0.15%) for a 4-week period. Theresults showed that enzyme supplementation improved growth performanceof piglets significantly (p < 0.05), but had no effect (p = 0.091)on average daily feed intake. The results also showed that supplementationof beta-glucanase and xylanase had no effect on pepsin activity in gastriccontents but slightly decreased (p = 0.092) the pepsin activity ingastric mucosa. Meanwhile, no effect of enzyme supplementation ontrypsin activity in duodenal contents was observed. However, the activitiesof amylase and lipase in duodenal contents were significantly(p < 0.05) decreased, whereas the activities of maltase, sucrase andgamma-glutamyl transpeptidase (gamma-GT) in jejunal and ileal mucosa wereenhanced significantly (p < 0.05). The improvement of disaccharidaseand gamma-GT activity may be attributed to the positive impacts of exogenousenzymes on digestion and absorption of the nutrients. In conclusion,the current results indicated that supplementation with enzymes in barley-based diets could improve the growth performance of piglets,decrease the activities of amylase and lipase in duodenal contents andincrease the activities of disaccharidase and gamma-GT in jejunal and ilealmucosa.

Objective: To study the expression of B7-H3 mRNA and B7-H3 protein in gastric carcinoma and their clinical significance., Methods: The expression of B7-H3 mRNA and B7-H3 protein in gastric carcinoma and the nearby normal tissue of 38 patients was detected by real-time RT-PCR and immunohistochemical assay respectively., Results: B7-H3 mRNA was expressed both in gastric carcinoma and nearby normal tissue, but the expression level in gastric carcinoma was much lower than that in nearby normal tissue. There were no significant differences of B7-H3 mRNA expression among gender, age, histological type, tumor size, lymph node metastasis and invasive depth (all P >0.05). The positive rate of B7-H3 protein expressed in gastric carcinoma was 39.5%. There were no significant differences of B7-H3 protein expression among gender, age, histological type, tumor size, lymph node metastasis and invasive depth (all P >0.05), but there were significant differences among groups of clinical stage (P=0.022) and pathological grade (P=0.039). Kaplan-Meier analysis revealed that disease-free survival or overall survival of the patients with positive B7-H3 expression were significantly longer than those with negative B7-H3 expression (P=0.009 and P=0.010 respectively)., Conclusion: Detection of B7-H3 expression in gastric carcinoma will be beneficial to the judgment of the prognosis of gastric carcinoma and the choice of individualized treatment.

Objective: To investigate the effects of Shenqi Fuzheng injection (SQFZI) combined with docetaxcl, flurouracil and calcium folinate in treating advanced gastric carcinoma, and to evaluate the action of SQFZI for enhancing therapeutic effect and alleviating adverse reaction of chemotherapy., Methods: Sixty advanced gastric cancer patients were assigned to two groups randomly, the control group treated with chemotherapy alone and the treated group treated with SQFZI combined chemotherapy. Chemotherapy regimens (TFC) was given to all patients consisting of docetaxel 75 mg/m2 intravenous dripping on the 1st day, flurouracil (5-FU) 350 mg/m2 and calcium folinate (CF) 120 mg/m2 intravenous dripping with micro-pump continuously from day 1 to 5, for 21-28 days as a cycle. To patients in the treated group, starting from 3 days before chemotherapy, SQFZI 250 mL was dripped every day for 14 successive days. The clinic effects were evaluated after two-cycle treatment., Results: The short-term effective rate was 40.0% in the treated group and 33.3% in the control group. As compared with those in the control group, patients in the treated group after treatment had a higher Karnofsky score (chi2 = 7.21, P < 0.05) and body weight (chi2 = 11.47, P < 0.05), lesser adverse reactions in decreasing of peripheral blood leucocyte, damage of peripheral nerve, adverse reaction of gastrointestinal tract, as well as better immune function., Conclusion: SQFZI could effectively improve the clinical symptoms induced by chemotherapy regimen TFC, alleviate the adverse reaction, raise patients' quality of life and their immune function.

After seedlings of Aeluropus sinensis var. sinensis Debeaux were treated with different NaCl concentrations (0-200 mmol/L) for 14 days, some physiological indexes were measured. The higher the NaCl concentration, the more the growth of A. sinensis was inhibited. The increase in root/shoot ratio suggests that the shoots are more sensitive to salinity than the roots. The diminished leaf area may reduce the transpiration rate, and the root mainly grew longitudinally, which may help the root to reach the water source under the high salinity conditions. Chl a contents increased, so did Chl b, but the Chl a/Chl b ratio declined, which implies the stimulation of Chl a accepted from NaCl is smaller than that of Chl b. After salt treatment, the organic solute contents increased (P<0.05), the proline, amino acids and soluble sugar contents increased more than organic acids, the increase in soluble carbohydrate may inhibit photosynthesis in feedback. The percentage of sucrose in soluble carbohydrates increased too. Although the organic dry weight of whole plant declined, the proportion of organic dry weight in total dry weight increased, and the osmotic potential of plant cell juice declined, which implies that the contribution of organic matter to osmotic adjustment increased with salinity. That is, A. sinensis had ability to tolerate salinity to a certain degree.

Aim: To construct and express a single chain fragment variable (scFv) fragment against human CD25 molecule and identify its bioactivity., Methods: V(H) and V(L) genes of anti-murine CD25 monoclonal antibody were cloned by RT-PCR from hybridoma cell WuTac secreting anti-CD25 mAb. scFv gene was spliced by sequence overlap extending (SOE) PCR, and then it was ligated into pMD18T vector to be identified by endonuclease digestion, PCR and sequencing. scFv gene was cloned into pBAD/gIIIA expression vector and transformed into TOP10 E.coli. After positive clones were induced by L-arabinose for 4 hours, the purity of protein was detected by SDS-PAGE and its bioactivity was identified by competitive inhibition ELISA test., Results: scFv genes of V(L)-(GGGGSGGGGSSGGGS)-V(H) was constructed successfully. The V(H) chain consisted of 351 bp and encoded 117 amino acids, which belonged to heavy chain subgroup III (C) of mouse immunoglobulin variable region. The V(L) chain consisted of 318 bp and encoded 106 amino acids, which belonged to light chain subgroup IV of mouse immunoglobulin variable region. The scFv antibody expressed by TOP10 fused with 6xHis and C-myc tag protein and the relative molecular mass of fusion protein was about 31,000. Competitive inhibition ELISA test indicated the scFv antibody had specific activity., Conclusion: The expressed product of the single-chain antibody shows some specific binding capacity, which provides a basis for the clinical application of anti-CD25 single-chain antibody.

The leaves of Aeluropus littoralis var. sinensis Debeaux were scanned with a scanning electron microscope, it showed that the upper-epidermis had almost the same number of salt glands as the lower-epidermis (Plate I-1, 2), and the salt gland is the typical bicelluar gland, which consists of a large basal cell inlaid into the epidermis and a small cap cell (Plate I-6). These salt glands were distributed mainly on the leaf veins, which favors the rapid collection of salts from the roots. Ion X-ray microanalysis indicated that the salt glands could effectively absorb Na(+) from the epidermal cells and mesophyllous cells (Table 1), then secreted Na(+) from the cap cells (Plate I-4), which would decrease the salinity of plant. After the plants were treated with various salts for 17 d, the ion contents of the leaves and the secretion were measured, and the results implied that salt glands had different selection in absorbing and secreting Na(+) , K(+) and Ca(2+), that is, the plants first selected K(+) when absorbing ions, while first selected Na(+) when sending ions out, but Ca(2+) was fewer in both absorbed and secreted, and the order of secretion of the three ions was found to be Na(+)>K(+)>Ca(2+) (Figs. 1-6). The secretion of Na(+) or three ions were respectively higher than that of leaves within 24 h (Figs. 1, 2, 7, 8), while K(+) situation was completely opposite with them (Figs. 3, 4). At the same time, the total ion content and composition inside the leaves remained more or less constant (Fig. 8).

Objective: To clone heavy-chain and light-chain variable region (VH and VL) gene of mouse-anti-human CD71 monoclonal antibody (mAb)., Method: One-step method was used to extract total RNA, and a set of oligonucleotide primers were designed to amplify the cDNAs with reverse transcriptase-polymerase chain reaction (RT-PCR), and the resultant products were respectively cloned into PMD18-T vector and their sequences analyzed., Results: The PCR product obtained with the oligonucleotide primers for the variable region of mouse immunoglobulin heavy chain was about 350 bp and that with oligonucleotide primers for the light chain was about 320 bp, and their DNA sequences were determined., Conclusion: The length of the cloned heavy chain variable region was 348 bp, belonging to mouse heavy-chain subgroup II(A); the light-chain variable region was 336 bp that belongs to mouse kappa light-chain subgroup II.

A selection of 16 monoclonal antibodies has been produced against a fresh Ewing's sarcoma (ES) tumor mixed with a permanent ES cell line. The majority of antibodies identify an 80-kDa molecule, which is not detected on healthy tissues except on certain cultured monocytes. One antibody recognizes the CD2 ligand MIC2 and 2 antibodies (numbers 13 and 16) define a higher-molecular-mass antigen. Antibody 16 is also expressed on mesenchymal fibroblasts of bone marrow or fetal origin. Tumor-specific antigen expression is potentially linked to the chromosome 22 abnormality described in Ewing's sarcoma, products altered expression in tumors with the chromosome 11/22 translocation has not been shown. The putative chimeric protein on chromosome 11 is apparently not expressed to a great extent, as tested by Northern blotting; however, the fusion protein initiated on chromosome 22 and ending on chromosome 11 is readily seen on Northern blots. The altered expression of a number of cellular genes in addition to a novel gene product(s) originating from translocation events were expected to be identified by monoclonal antibodies selected by their unique binding pattern to Ewing's sarcoma (ES) cells.

Pokeweed antiviral protein (PAP-s) was prepared from seeds of Phytolacca americana. Monoclonal antibody against human pan-T lymphocyte Wu71 was linked to PAP-s by a disulfide bond. The results of SDS-PAGE, double immunodiffusion of active monoclonal antibody and PAP-s showed that the conjugate was highly cytotoxic to the human T-leukemic cell line CEM, but not to antigen-negative cell line SP2/O. At a concentration of 10(-9) mol/L, 76.4% of the target cells were killed, as compared with 10.1% at 10(-9) mol/L of free PAP-s. Treatment of the CEM cells with conjugate at 10(-9) mol/L reduced their rate of protein synthesis by 72.4%, as determined with 14C-leucine incorporation. The immunotoxin may be useful for the in-vitro eradication of leukemic cells in autologous bone marrow transplantation to leukemia patients.

The specificity of a novel monoclonal antibody (moAB), TU69, directed to the interleukin 2 receptor (IL-2R) was verified by sequential immunoprecipitation with anti-Tac. TU69 cross-competed with anti-Tac in binding analyses. When TU69 was added during the sensitization of normal peripheral blood mononuclear cells (PBMC) to allogeneic HLA-class I or -class II mismatched stimulator PBMC, alloproliferative responses and specific cytotoxicity were no longer detectable and the generation of natural killer (NK)-like effector cells was partially inhibited. Remarkably, however, the generation of CD4+ nonspecific suppressor T cells in such mixed lymphocyte cultures (MLC) was not inhibited--but, in contrast, was strongly enhanced in the presence of TU69. These suppressor cells inhibited unrelated allospecific responses in vitro to background levels even at a ratio of 50:1 responder:irradiated suppressor T cell lines. Such a potent experimental suppressor system suggests a possible application of TU69 for in vivo tolerance induction after transplantation, by down-regulating allospecific effector cells and allowing the generation of tolerance to graft antigens.