Objective To investigate the effects of exosomal microRNA-1234-3p(miR-1234-3p)of lung cancer cells on the malignant behaviors of lung cancer cells,the polarization of tumor-associat-ed macrophages(TAMs),and the functions of T cells,as well as its potential molecular mecha-nisms.Methods Exosomes were extracted from lung cancer A549 cells,and their morphology was observed using transmission electron microscopy.The expression of exosomal marker proteins[tumor susceptibility gene 101(TSG101),CD63 and CD9]was detected by western blot.The relative ex-pression level of miR-1234-3p was measured using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The expression of exosomal miR-1234-3p was inhibited by transfecting,the proliferation,migration,and invasion abilities of A549 cells were detected using the CCK-8 method,wound healing assay,and Transwell invasion assay,respectively.The exosomes were co-cultured with macrophages and T lymphocytes separately.The expression of TAM polarization markers CD206 and CD163 was detected by western blot.The secretion levels of cytokines[interleukin(IL)-6,IL-10,and transforming growth factor-β(TGF-β)]were measured using an enzyme-linked immu-nosorbent assay(ELISA).The proliferation and apoptosis abilities of T lymphocytes,as well as the expression of functional molecules[γ-interferon(IFN-γ)and programmed death receptor 1(PD-1)]were detected through cell experiments.The expression levels of proteins related to the phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(AKT)signaling pathway were detected by western blot.Results The isolated exosomes were round double-layer vesicles with a diameter of 50 to 200 nm and expressed TSG101,CD63 and CD9 proteins.The expression level of miR-1234-3p in A549 cells was higher than that in human normal lung epithelial cells BEAS-2B,and the expression level of miR-1234-3p in A549 cell exosomes was higher than that in parental A549 cells,with statis-tically significant differences(P＜0.001).After inhibiting exosomal miR-1234-3p,the prolifera-tion,migration,and invasion abilities of A549 cells decreased,with statistically significantdifferenc-es(P＜0.001).After inhibiting exosomal miR-1234-3p,the levels of M2-type polarization marker proteins CD206 and CD163 in macrophages decreased,the levels of cytokines IL-10 and TGF-β de-creased,and the level of IL-6 increased,with statistically significant differences(P＜0.001),indi-cating that the polarization of macrophages toward the M2 type was inhibited.In T lymphocytes,af-ter inhibiting exosomal miR-1234-3p,cell viability increased,the apoptosis rate decreased,the ex-pression level of the functional molecule IFN-γ increased,and the expression level of PD-1 de-creased,with statistically significant differences(P＜0.01 orP＜0.001).After inhibiting exosom-al miR-1234-3p,the protein levels of phosphorylated(p)-PI3K and p-AKT in A549 cells de-creased,with statistically significant differences(P＜0.001).Conclusion Exosomal miR-1234-3p of lung cancer cells may promote the malignant behaviors of lung cancer cells,the polarization of TAMs toward the M2 type,and the dysfunction of T lymphocytes by activating the PI3K/AKT signa-ling pathway,thereby regulating the immunosuppressive microenvironment and promoting the pro-gression of lung cancer.

ObjectiveTo observe the effect of tendon clearing combined with Zizhu ointment on the serum levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） inflammasomes， interleukin （IL）-1β， and IL-18 in the treatment of non-ischemic diabetic foot ulcers. MethodA total of 106 patients with non-defective diabetic foot ulcers who attended the outpatient clinic and wards of the Vascular Surgery Department of Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine from March 2023 to March 2024 were selected. The patients with non-ischemic diabetic foot ulcers who met the inclusion criteria were assigned with the random number table method into an observation group and a control group， with 53 patients in each group. Patients in both groups received basic treatment. The local ulcers in the observation group received tendon clearing combined with Zizhu ointment， while those in the control group received conventional debridement combined with topical solution of bovine basic fibroblast growth factor. The serum NLRP3， IL-1β， and IL-18 levels， ulcer area， traditional Chinese medicine （TCM） symptom scores， visual analogue scale （VAS） score， and DMIST score were measured and recorded in the two groups before and after treatment. The ulcer healing rate and incidence of adverse reactions were compared between the two groups. ResultThe observation group had higher ulcer healing rate than the control group （P<0.01）. After treatment， both groups showed lowered serum NLRP3， IL-1β， and IL-18 levels， reduced ulcer area， and declined TCM symptom score， VAS score， and DMIST score （P<0.01）. Moreover， these indicators were lower in the observation group than in the control group （P<0.05， P<0.01）. Neither group showed significant changes in the liver and kidney function indicators after treatment. Significant adverse reactions occurred in neither group during the treatment course. ConclusionTendon clearance combined with Zizhu ointment was effective and safe in treating non-ischemic diabetic foot ulcers. It may exert the therapeutic effect by reducing the inflammation of the local ulcers.

BACKGROUND: Corneal confocal microscopy (CCM) is a noninvasive technique to detect early nerve damage of diabetic sensorimotor polyneuropathy (DSPN). Time in range (TIR) is an emerging metric of glycemic control which was reported to be associated with diabetic complications. We sought to explore the relationship between TIR and corneal nerve parameters in asymptomatic patients with type 2 diabetes (T2DM). METHODS: In this cross-sectional study, 206 asymptomatic inpatients with T2DM were recruited. After 7 days of continuous glucose monitoring, the TIR was calculated as the percentage of time in the glucose range of 3.9 to 10.0 mmol/L. CCM was performed to determine corneal nerve fiber density, corneal nerve branch density, and corneal nerve fiber length (CNFL). Abnormal CNFL was defined as ≤15.30 mm/mm 2 . RESULTS: Abnormal CNFL was found in 30.6% (63/206) of asymptomatic subjects. Linear regression analyses revealed that TIR was positively correlated with CCM parameters both in the crude and adjusted models (all P   <  0.05). Each 10% increase in TIR was associated with a 28.2% (95% CI: 0.595-0.866, P  = 0.001) decreased risk of abnormal CNFL after adjusting for covariates. With the increase of TIR quartiles, corneal nerve fiber parameters increased significantly (all P for trend <0.01). The receiver operating characteristic curve indicated that the optimal cutoff point of TIR was 77.5% for predicting abnormal CNFL in asymptomatic patients. CONCLUSIONS There is a significant independent correlation between TIR and corneal nerve fiber loss in asymptomatic T2DM patients. TIR may be a useful surrogate marker for early diagnosis of DSPN.
Humans ; Diabetes Mellitus, Type 2/complications* ; Cross-Sectional Studies ; Blood Glucose Self-Monitoring ; Blood Glucose ; Nerve Fibers ; Diabetic Neuropathies ; Cornea ; Microscopy, Confocal/methods*

Objective To summarize the experiences of aneurysmorrhaphy for arteriovenous fistula aneurysms with acute thrombosis in hemodialysis patients.Methods There were 7 cases of arteriovenous fistula with acute thrombosis from Nov 2015 to Feb 2017 at our department of vascular surgery,Longhua Hospital.Results In all cases thrombosis was secondary to proximal stenosis or occlusion.The stenosis and occlusion were corrected with embolectomy and aneurysmorrhaphy.The proximal part of the cephalic vein was translocated to the basilic vein in 1 case,axillary vein in 2 cases;autologous vein graft in 1 case;resection of the occlusion,end-to-end anastomosis in 1 case;autogenous patch in 1 case.No perioperative complications occurred.The operation site was cannulated within one month after operation in all cases.Patients were followed up for 7 months to 23 months,all cases were patent.Conclusions Aneurysmorrhaphy is effective,reliable and safe for arteriovenous fistula aneurysms with acute thrombosis in hemodialysis patients.

Objective To observe the clinical efficacy ofXing Nao Kai Qiao(brain-awakening orifice-opening) needling method plus Frenkel's balance rehabilitation exercise in treating lower-limb dysfunction due to different types of ataxia after stroke.MethodA total of 115 eligible patients were randomized into two groups. Fifty-seven cases in the treatment group were intervened by Xing Nao Kai Qiaoneedling plus rehabilitation training; 58 cases in the control group were intervened byXing Nao Kai Qiaoneedling method alone. Berg Balance Scale (BBS) was adopted to evaluate the symptoms, balance function, and therapeutic efficacy before and after the treatment.Result In the treatment group, the markedly effective rate was respectively 72.2%, 83.3% and 61.1% in the cerebella subtype, brainstem subtype and basal ganglia subtype, and the total effectiverate was respectively 100.0%, 94.4% and 94.4%; in the control group, the markedly effective rate was respectively 21.1%, 35.0% and 58.8%, and the total effective rate was respectively 94.7%, 85.0% and 82.4%, and the between-group differences were statistically significant (P<0.05). After the treatment, the BBS score showed significant intra-group differences in both groups (P<0.01); the between-group comparisons showed that there were significant differences in comparing the scores of cerebella and brainstem subtypes after the treatment (P<0.05).ConclusionXing Nao Kai Qiaoneedling method plus Frenkel's rehabilitation exercise can effectively improve the symptoms of poststroke lower-limb ataxia, especially for cerebella and brainstem subtypes.

To construct soluble TNF related apoptosis inducing ligand (TRAIL) expression system and investigate the effect of the expression product on tumor cell. It may provide valuable information for research into the immune system of the finless porpoise. The full-length cDNA of TRAIL (designated fTRAIL) was cloned from the total RNA of the finless porpoises blood using RT-PCR techniques and then the extracellular soluble fragments of fTRAIL (designated fsTRAIL) was ligated into pET43.1a. Recombinant soluble fTRAIL (pET43.1a-fsTRAIL) fused with Nus-his tag was efficiently expressed in Escherichia coli BL21 (DE3) and the Nus-His-fsTRAIL protein was purified. The expression of Nus-His-fsTRAIL was verified by Western blotting. In vitro, the effects of the purified Nus-His-fsTRAIL protein on Jurkat and HeLa cells were etected by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide (MTT) assay, TrypanBlue and Flow Cytometry analysis. The expression system pET43.1a-fsTRAIL was constructed and Nus-His-fsTRAIL protein was expressed successfully. In vitro, the Nus-His-fsTRAIL protein was able to inhibit the proliferation and induce apoptosis of Jurkat and HeLa cells in a dose-dependent manner. The Nus-His-fsTRAIL protein has anti-tumor activity against Jurkat and HeLa cells in vitro.
Animals ; Apoptosis ; Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; HeLa Cells ; Humans ; Jurkat Cells ; Porpoises ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis

Objective To analyze the clinical characteristics and recent curative effect of mantle cell lymphoma (MCL) after conventional treatment.Methods Clinical data of 15 MCL patients admitted in the Affiliated Hospital of Academy of Military Medical Sciences between August 2004 and October 2013 were retrospectively analyzed.Results The median age of those patients was 59 and the male to female ratio was 1.5∶1.Fourteen(93%)cases were in Ann-Arbor stages Ⅲ -Ⅳ, 15 cases (100%)primarily with lymph node involvement,7 cases (47%)with bone marrow involvement,4 cases (27%)with gastrointestinal involvement,and 3 cases (20%)with orbit involvement.Less than 40% expression of Ki-67 was observed in 9 cases (60%),while 6 cases were with more than 40% (40%).One case was blastic variant.First-line therapy was CHOP-like regimens,which were combined with rituximab in 8 of the 15 cases.In this study,the median survival time was 12 months (3 -64),and the overal response rate was 80%after induction chemotherapy.The current survival of 7 /9 cases with less than 40% expression of Ki-67 was 8 -64 months,2 /6 cases with more than 40% expression of Ki-67 was 8 and 9 months,respectively.Conclusion MCL mostly occurs in older males.Extranodal invasion is common in MCL as an aggressive tumor.The efficacy of traditional chemotherapy is currently limited.Blastic variant or high expression of Ki-67 is an adverse prognostic indicator.

Objective:The effect and side effect of the dose-adjusted EPOCH regimen were evaluated perspectively for the pre-liminarily diagnosed angioimmunoblastic T-cell lymphoma. Methods: Nine cases of untreated angioimmunoblastic T-cell lymphoma were diagnosed and enrolled in our department from September 2008 to September 2012. All patients received dose-adjusted EPOCH regimen as first-line chemotherapy. Results: The median age of 9 patients was 54 years. The male-to-female ratio was 2∶1. About 88.9%of all patients were at Ann Arbor stageⅢ/Ⅳ, and 77.8%presented with B symptoms. Anemia was found in 66.7%of 9 patients, and lactate dehydrogenase elevated in 55.6%of patients. After an average of 4.7 cycles of chemotherapy of dose-adjusted EPOCH regi-men, the complete remission rate was 22.2%, and the total response rate was 66.7%. With a median follow-up of 20 months, the 4-year progression-free survival rate was 11.1%, and the overall survival rate was 33.3%. The median survival time was 19 months. The most common adverse events of EPOCH chemotherapy were hematologic toxicity. Grades 3-4 neutropenia and thrombocytopenia were re-ported in 77.8%and 33.3%of patients. Febrile neutropenia was observed in 44.4%of patients. Non-treatment-related mortality was al-so noted. Conclusion: The results of our research showed no clear benefit of treating preliminarily diagnosed angioimmunoblastic T-cell lymphoma with dose-adjusted EPOCH regimen. The main adverse events were hematologic toxicity and could be tolerated.

Objective To study the early and mid-term outcome in terms of left ventricular(LV)geometry and function in patients with transposition of great arteries with intact ventricular septum (TGA/IVS) undergoing arterial switch operation.Methods Twenty patients were followed up and divided into 2 groups according to age:the earlier group and the later group.Age and sexual matched controls were also designed as control group 1 and control group 2.LV posterior wall thickness(PWTd),dimension(LVIDd)and length(L) at end diastolic phase,LV end-diastolic volume (EDV),end-systolic volume (ESV),ejection fraction (EF),mitral valve early (E) and late (A) inflow velocities and E/A,isovolumic relaxation time (IVRT) were obtained by two-dimensional echocardiography,normalized isovolumic relaxation time (IVRTn),wall thickness index (PWTd/LVIDd) and LV geometry index(LVIDd/L) were calculated,and these indexes between groups were compared.Results ①Compared with control group 1,PWTd in earlier group was higher [(0.41 + 0.06)cm vs (0.36 + 0.05)cm,P =0.022],but LVIDd had no significant statistical difference [(2.08 + 0.21)cm vs (2.21 + 0.23)cm,P =0.117],indicating PWTd/ LVIDd differed between groups(0.20 + 0.04 vs 0.16 + 0.01 P =0.001).②No abnormalities were observed in later group.③Compared with different age groups in TGA,EDV,ESV,L,LVIDd,PWTd were significantly higher in later group(P =0.000).There were obvious correlations between EDV,ESV,L,LVIDd and examined age (r =0.90,0.83,0.73,0.90,P =0.000,R2 =0.809,0.694,0.528,0.808) while there were no significant correlation between PWTd and examined age(r =0.56,P =0.000,R2 =0.309).Conclusions Early after operation,LV is undergoing hypertrophy or hyperplasia in TGA/IVS,but will recover in the mid-term period,and the function is normal all the time indicating that the LV myocardium may avoided irreversible pathological changes if operated in time.All in all,LV develops well in mid-term period.

Objective To isolate and identify leukemia stem cells from acute myeloid leukemia patients for further research. Methods By Ficoll density gradient centrifugation, mononuclear cells were firstly separated from bone marrow of patients. According to specific surface markers, CD+34 CD+123 of leukemic stem cells were sorted by flow cytometer. Their ability of self-renewal and differentiation were evaluated by colony formation and cobblestone forming ability. At the same time, the purity and cell morphology of CD+34 CD+123 cells was analysed. Results Comparared with total mononuclear cells, the proportion of the CD+34 CD+123 cells after sorting was 10.7 %, and these cells showed the ability of colony forming and cobblestone forming, and the purity proportion of CD+34 CD+123 cells was 62.1%. Conclusion The leukemia stem cells were isolated successfully and could be used in further study.