Objective Zinc finger protein 335 (Zfp335) plays a crucial role in the early development of thymic T cells and the differentiation of peripheral T cell subpopulations. The objective of this study is to investigate the role and underlying mechanisms of Zfp335 in the regulation of regulatory T cell (Treg) within tumor immunity. Methods The Zfp335 gene was specifically knocked out in Treg using tamoxifen (Zfp335^fl/fl FOXP3^creERT2), and the MC38 tumor model was established. On the 7th day after tumor inoculation, tumor size was observed and measured. Tumor size was monitored and recorded daily starting from day 7 post-inoculation. On day 12, tumors were harvested, and the proportions of CD4⁺ T cells, CD8⁺ T cells, and Treg were analyzed by flow cytometry. Additionally, the mitochondrial function of effector regulatory T cell (eTreg) was assessed. Results From day 10 post-tumor inoculation, tumor volume in the Zfp335^CKO group was significantly reduced compared to that of the wild-type (WT) group. Furthermore, the infiltration of CD4⁺ and CD8⁺ T cells, along with their respective effector cells, was significantly higher in the Zfp335^CKO group than in the WT group. The proportions of CD4⁺ and CD8⁺ T cells producing interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were also significantly increased in the Zfp335^CKO group compared to that of the WT group. In addition, the percentage of CD8⁺ T cells secreting granzyme B (GzmB) was significantly higher in the Zfp335^CKO group than that in the WT group. In contrast, the proportion of Treg and inducible T cell co-stimulator (ICOS)⁺ Treg in the Zfp335^CKO group was significantly lower than that in the WT group. Finally, the expression level of Mitotracker Deep Red in eTreg from the Zfp335^CKO group was significantly reduced compared to that in the WT group. Conclusion During tumorigenesis, the specific deletion of Zfp335 impairs Treg activation, which is related to decreased mitochondrial function in eTreg. In Zfp335^CKO mice. Tumors exhibit increased infiltration of effector T cells, accompanied by elevated levels of cytotoxic cytokines, ultimately enhancing resistance to tumor progression.
Animals ; T-Lymphocytes, Regulatory/metabolism* ; Mice ; CD8-Positive T-Lymphocytes/immunology* ; Neoplasms/genetics* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice, Knockout ; DNA-Binding Proteins/genetics* ; Female

ObjectiveThis study aims to investigate the changes in fungal community diversity and volatile components during the aging process of Aquilariae Lignum Resinatum and explore the internal relationship between them. MethodAquilariae Lignum Resinatum samples with different aging years were collected. High-throughput sequencing was employed to analyze the fungal diversity and abundance， and α and β diversity indicators were calculated to reveal the composition and dynamic changes of the fungal community. In addition， the essential oils of Aquilariae Lignum Resinatum with different aging years were extracted， separated， and identified by two-dimensional gas chromatography-high resolution time-of-flight mass spectrometry. ResultA total of 61 compounds were identified from the volatile components of five groups of Aquilariae Lignum Resinatum samples， including 2 monoterpenes， 24 sesquiterpenoids， 1 diterpene， 13 aromatic hydrocarbons， 9 alkanes， and 12 other compounds. Among them， the volatile compounds isolated from the sample aged for 1 year had the largest number， and those from the sample aged for 2 years accounted for the largest proportion of the total components. The internal transcribed spacer（ITS） amplicon sequencing revealed that the fungi in the five groups of samples belonged to 162 genera. Kirschsteiniothelia， Aspergillus， Lasiodiplodia， Phaeoacremonium， and Trichoderma were the dominant fungal genera. The fungal diversity in the Aquilariae Lignum Resinatum sample aged for 4 years was significantly higher than that in the samples aged for 0 to 3 years. ConclusionThe volatile component content and composition of Aquilariae Lignum Resinatum altered dramatically during aging. The aging of Aquilariae Lignum Resinatum was accompanied by the increasing fungal diversity， decreasing relative content of aromatic hydrocarbons， and increasing relative content of sesquiterpenoids. In general， aging was beneficial to the transformation of sesquiterpenoids and the enrichment of fungi.

【Objective】 To evaluate the effect of Arntl on T cell development and T cell-mediated anti-viral immunity. 【Methods】 Arntl^F/FCD4cre⁺(KO) in mice was constructed to delete Arntl gene specifically in T cells. We examined the percentage and number of T cell subsets in the thymus and spleen by flow cytometry (FCM). At day 8 after lymphocytic choriomeningitis virus (LCMV) infection, the proportions of T cell subsets, virus-specific CD8⁺ T cells and IFN-γ secreting T cells were analyzed. The viral load in the spleen was measured using qPCR. Naive CD4⁺ T cells (CD4⁺CD25^-CD44^-CD62L⁺) were sorted by flow cytometry to perform T helper cell differentiation in vitro. 【Results】 The percentage and number of T cells in the thymus and spleen of KO mice showed no significant change compared with those in the control group (Arntl^F/FCD4cre^- mice, WT) (P>0.05). Acute LCMV infection did not cause observable changes in effector T cell proportion in the spleen of KO mice compared to that in WT mice (P>0.05), but KO mice showed a higher proportion of IFN-γ secreting T cells (P<0.05) and better virus clearance (P<0.05). In addition, naive CD4⁺ T cells from KO mice were more prone to differentiate into Th1 cells in vitro (P<0.05). 【Conclusion】 Arntl deletion in T cells does not affect T cell development, but enhances their ability to defend against viral infection by promoting Th1 cell differentiation and response.

Objective To evaluate the effects of nasolabial flap with facial artery and its branches perforator for reconstruction of nasal defect.Methods Between March 2013 and April 2017,21 patients underwent operations for the reconstruction of nasal defect,caused by trauma,surface tumors,moles and infection.The size of the defect was 1.5 cm × 2.0 cm to 3.0 cm × 3.0 cm.Designed various nasolabial perforator flap was pedicled with the facial artery.The pulsed blood flow detector determined the location of the facial artery and its perforation position,which was the rotation point,and the rotation of the nasolabial fold flap covered the nasal defect area to repair.Results 21 flaps survived.Surface artery perforation nasolabial fold flap was good blood supply,of which 1 case of flap was congested and recovered after treatment.After 1 month to 3 years follow-up on 21 cases,20 cases showed good results and 1 case had generally accepted.The color,shape and function of the flap were significant,similar to the normal skin.Conclusions A small area defect in the nose is preferred by using facial arterial perforation nasolabial fold flap repair,which does not need secondary repair,and is worthy of clinical application.

Three new lignan glucosides, baicalensinosides A-C (1-3), were isolated from the roots of Scutellaria baicalensis. The structural elucidation was achieved by in-depth spectroscopic examinations and qualitative chemical test. Structurally, these compounds belong to the 3,4-dibenzyltetrahydrofuran-type lignan glycoside with a mono-hydroxyl substitution at the 7'-position of benzylidene group on the numbering system of lignans being one of their shared critical features. The anti-osteoporotic activity of the isolated compounds was assessed in an in vitro osteoprotegerin (OPG) transcriptional activity assay using dual luciferase reporter detection. At 10 μmol/L, compounds 1-3 increased the relative activating ratio of OPG transcription to 1.83, 0.84 and 0.98 times that of the control group, respectively.

Ongoing study on the chemical constituents of the roots of Macleaya microcarpa led to the isolation of eight compounds of derivatives of triterpenes and organic acids in addition to some previously identified benzophenanthridines. The eight compounds were identified by spectroscopic methods as well as comparison with literature values as 1-oxo-2, 22 (30)-hopandien-29-oic acid (1), 3-oxo-12-oleanen-30-oic acid (2), 3α-hydroxy-12-oleanen-30-oic acid (3), 3β-hydroxy-12-oleanen-30-oic acid (4), ferulic acid (5), ferulic acid 4-O-β-D-glucoside (6), 3-O-feruloylquinic acid (7), and methyl 3-O-feruloylquinate (8). Of which, 1 is a new triterpenoid of hopanes and 2-8 are isolated from M microcarpa for the first time. In order to discover natural active compounds as potential agents of anti-ulcerative colitis (UC), an in vitro drug high-throughput screening model targeted x-box-binding protein 1 (xbp1) was employed to evaluate the activity of the major chemical constituents of M microcarpa. The result confirmed that two dihydrobenzophenanthridines, dihydrosanguinarine (9) and dihydrochelerythrine (10), showed a certain activity on activating the transcription of xbpl, a transcription factor (TF) associated with the occurrence, development, and potential treatment of UC, with their relative activating ratios being 1.76 and 1.77 times, respectively, as compared with control group.

Objective The aim of this study was to establish rat models of immunosuppression and immune hyper-function.Methods Sixty-four SPF Sprague-Dawley rats were randomly divided into groups A, B, and C.All rats were immunized with intraperitoneal injection of 100 μg ovalbumin ( OVA) .The group A was used as control.At 6 hours after immunization, the rats of group B were injected with different doses of cyclophosphamide (Cy) at different time points.The rats of group C were injected with Cy in different ways at 3 days before immunization.Results Immunosuppressed rats were successfully induced by Cy (125 mg/kg or 100 mg/kg) at 6 h after immunization and also by injection of 225 mg/kg Cy at 3 days before immunization with ovalbumin.Small dose (20 mg/kg) of Cy injected once or a smaller dose (5 mg/kg/d) injected once a day for consecutive 3 days can also result in immune hyperfunction.Conclusions Rat models of immunosuppression and immune hyperfunction are successfully established, which provide methodological and data support for establishment of such animal models and useful reference for related research.

OBJECTIVETo investigate the cytotoxicity of normal CD8(+) T lymphocytes retrovirally transduced with WT1 peptide-specific T-cell receptor (TCR) genes against human lung cancer cells.

METHODSHLA-A*2402-restricted and WT1 peptide-specific TCR-α/β genes were cloned from a cytotoxic T lymphocyte clone and inserted into a retroviral TCR expression vector. The cytotoxicity of normal peripheral CD8⁺ T cells transduced with the WT1-TCR genes against human lung cancer cells was evaluated using a standard ⁵¹Cr release assay.

RESULTSThe WT1-TCR gene-modified T cells recognized the peptide-pulsed target cells but not the non-pulsed cells. TCR-redirected CD8⁺ T cells lysed WT1-overexpressing human lung cancer cells in an HLA-A*2402-restricted manner, but did not kill normal cells positively expressing HLA-A*2402.

CONCLUSIONThese data demonstrate the feasibility of adoptive immunotherapy with TCR-redirected T cell for the treatment of lung cancer.

CD8-Positive T-Lymphocytes ; cytology ; Cell Line, Tumor ; Genes, T-Cell Receptor ; Humans ; Immunotherapy, Adoptive ; Lung Neoplasms ; pathology ; Peptides ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; Retroviridae ; T-Lymphocytes, Cytotoxic ; cytology ; Transduction, Genetic ; WT1 Proteins ; genetics

Objective To investigate the glycosidic linkage and immunomodulating activities of crude polysaccharides from the seeds of Zea mays. Methods Extraction, isolation and purification of polysaccharides were carried out with boiling-water extraction plus resolving deposit repeatedly. The structure was elucidated on basis of physicochemical properties and spectral data, and the biological activities were evaluated by means of Immunopharmacological examination. Results The structure of polysaccharides from the seeds of Z. mays exhibited identical structure with rice bran polysaccharides, i.e., a kind of glucan withα-1,4 andα-1,6 glucosidic bonds as the main frame. Conclusion Polysaccharides was obtained from the seeds of Z. mays for the first time, and it showed significant immunomodulating activity in mice.

Objective:To study the protective effects of collagen peptide-chromium (Ⅲ) complex (CPCC) on alloxan-induced hepatic injury in mice. Method:The mice were divided into three groups (normal, model, CPCC) and given (po.) water, water and CPCC (Cr3+40?g/kg?d) respectively once a day. After 4 weeks, the model and CPCC groups were injected with alloxan. Then the levels of liver index, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) were measured. Activities of superoxide dismutase (SOD) ,glutathione peroxidase (GSH-Px) and contents of malondialdehyde (MDA) in liver were tested. Also liver pathological changes were observed. Results:CPCC could reduce the levels of liver index, serum levels of AST,ALT and ALP as well as contents of MDA and activities of SOD and GSH-Px in liver. Hepatocytes lesion was alleviated markedly. Conclusion:CPCC has protective effects on alloxan-induced hepatic injury in mice.