BACKGROUND:M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing.Therefore,how to regulate M2 polarization of macrophages has been a hot research topic in recent years,and some miRNAs have been found to have this function. OBJECTIVE:To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line. METHODS:The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ.Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages.By transfection with lentivirus as the vector of overexpression of miR-378a,the stable expression of miR-378a cell lines was screened.Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ.Macrophage M2 polarization was induced by interleukin-4.The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay.qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. RESULTS AND CONCLUSION:(1)The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization.(2)Compared with the non-transfected group,the expressions of proinflammatory cytokine-induced nitric oxide synthase,tumor necrosis factor-α,interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group(P<0.05);the levels of inducible nitric oxide synthase,tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased(P<0.05).(3)Compared with the non-transfected group,the expressions of CD206,interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased(P<0.05);the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased(P<0.05)in the miR-378a transfection group.(4)It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages.

Objective To compare the early embryonic developmental potential and clinical outcomes of oocytes matured in vivo and those matured by modified in vitro maturation(LVM)technology during the same controlled ovarian hyperstimulation(COH)cycle,and to explore the clinical application of melatonin-supplemented IVM technology.Methods 159 patients were recruited into the study.920 mature oocytes were collected during their COH cycles processed for conventional IVF/ICSI protocols,while 1 283 immature oocytes from the same cycles were matured in a melatonin-supplemented IVM medium before ICSI was performed.A retrospective analysis was conducted to compare the impact of conventional assisted reproductive technology and improved IVM technology on the outcomes of assisted reproductive therapy and pregnancy outcomes.Results Compared with mature oocytes collected from COH cycles treated with conventional IVF/ICSI,oocytes promoted by improved melatonin-supple-mented IVM technology had a lower rate of high-quality blastocyst formation.However,after embryo transfer,there was no significant difference in the clinical outcomes of mature oocytes obtained through two methods,including clinical pregnancy rate,full-term birth rate,neonatal length,and neonatal Apgar score.Conclusion The applica-tion of melatonin-supplemented IVM significantly increases the utilization of immature oocytes collected from COH cycles,improving the pregnancy outcomes of patients assisted by assisted reproductive technology.

Objective To investigate the mechanism underlying the inhibitory effects of Demethylzeylasteral(T-96)on non-small cell lung cancer(NSCLC)cells.Methods We first examined the effects of different concentrations(1,3,10,and 30 μmol/L)of demethylzeylasteral on morphology and cell number of A549 and H1299 cells.The changes in proliferation,cell viability,migration,invasion,and apoptosis of A549 and H1299 cells following demethylzeylasteral treatment were detected using clone formation,CCK-8,cell scratch,Transwell,and flow cytometric assays,and the effect of SC79 treatment against demethylzeylasteral-induced cell apoptosis was assessed.Western blotting was performed to detect the changes in expressions of E-cadherin,N-cadherin,vimentin,Bax,Bcl-2 and cleaved caspase-3 and phosphorylation of AKT/CREB in demethylzeylasteral-treated A549 and H1299 cells and the cellular expressions of apoptotic proteins following treatment with both demethylzeylasteral and SC79.Results T-96 treatment caused elongation of the cell body and widening of the intercellular space and significantly inhibited cell viability,proliferation,migration and invasion of A549 and H1299 cells(P<0.05).Flow cytometry showed that demethylzeylasteral induced apoptosis in both A549 and H1299 cells,whereas SC79 treatment obviously attenuated its pro-apoptotic effect(P<0.05).Western blotting revealed up-regulated expressions of Bax and cleaved caspase-3 proteins and lowered Bcl-2 expression level in demethylzeylasteral-treated A549 and H1299 cells,but co-treatment with SC79 obviously attenuated the expressions of the apoptotic proteins.T-96 significantly up-regulated the expression level of E-cadherin,down-regulated the expressions of N-cadherin and vimentin,and inhibited the phosphorylation of AKT and CREB in the two cell lines(P<0.05).Conclusion T-96 inhibits the proliferation,migration and invasion and induces apoptosis of NSCLC cells possibly by inhibiting the AKT/CREB signaling pathway.

Objective To investigate the role of miR-194-3p in regulating functional changes in osteoblasts in a simulated microgravity environment and to provide a theoretical foundation for understanding the mechanical response mechanisms of osteoblasts in extreme mechanical environments.Methods The effects of microgravity on osteoblasts were simulated by using a rotary cell culture system.MC3T3-E1 osteoblasts were transfected with an miR-194-3p inhibitor and changes in proliferation,differentiation,apoptosis,and mineralization were assessed using MTT assay,RT-PCR,Western blot,double fluorescence staining,and alizarin red staining.Results Elevated expression of miR-194-3p under simulated microgravity conditions led to the suppression of osteoblast proliferation,differentiation,and mineralization to a certain extent,while promoting osteoblast apoptosis.However,transfection with the miR-194-3p inhibitor significantly downregulated miR-194-3p expression and partially reversed the reduced osteoblast proliferation,decreased expression of osteogenic differentiation markers such as ALP,OCN,and COL-I genes and proteins,decreased bone mineralization nodules,and increased osteoblast apoptosis induced by microgravity exposure.These findings indicated that miR-194-3p effectively ameliorates abnormal osteoblast function under microgravity conditions.Conclusions MiR-194-3p acts as a negative regulatory factor in the mechanical responses of osteoblasts under simulated microgravity.

Objective To investigate the mechanism underlying the inhibitory effects of Demethylzeylasteral(T-96)on non-small cell lung cancer(NSCLC)cells.Methods We first examined the effects of different concentrations(1,3,10,and 30 μmol/L)of demethylzeylasteral on morphology and cell number of A549 and H1299 cells.The changes in proliferation,cell viability,migration,invasion,and apoptosis of A549 and H1299 cells following demethylzeylasteral treatment were detected using clone formation,CCK-8,cell scratch,Transwell,and flow cytometric assays,and the effect of SC79 treatment against demethylzeylasteral-induced cell apoptosis was assessed.Western blotting was performed to detect the changes in expressions of E-cadherin,N-cadherin,vimentin,Bax,Bcl-2 and cleaved caspase-3 and phosphorylation of AKT/CREB in demethylzeylasteral-treated A549 and H1299 cells and the cellular expressions of apoptotic proteins following treatment with both demethylzeylasteral and SC79.Results T-96 treatment caused elongation of the cell body and widening of the intercellular space and significantly inhibited cell viability,proliferation,migration and invasion of A549 and H1299 cells(P<0.05).Flow cytometry showed that demethylzeylasteral induced apoptosis in both A549 and H1299 cells,whereas SC79 treatment obviously attenuated its pro-apoptotic effect(P<0.05).Western blotting revealed up-regulated expressions of Bax and cleaved caspase-3 proteins and lowered Bcl-2 expression level in demethylzeylasteral-treated A549 and H1299 cells,but co-treatment with SC79 obviously attenuated the expressions of the apoptotic proteins.T-96 significantly up-regulated the expression level of E-cadherin,down-regulated the expressions of N-cadherin and vimentin,and inhibited the phosphorylation of AKT and CREB in the two cell lines(P<0.05).Conclusion T-96 inhibits the proliferation,migration and invasion and induces apoptosis of NSCLC cells possibly by inhibiting the AKT/CREB signaling pathway.

ObjectiveTo observe the clinical efficacy of Chinese medicine combined with indirect moxibustion plaster on corona virus disease 2019 （COVID-19） patients during recovery period. MethodNinety patients of COVID-19 during the recovery period were randomly divided into a Chinese medicine group， an indirect moxibustion plaster group， and a combination group，with 30 cases in each group. According to the 10th edition of COVID-19 Diagnosis and Treatment Protocol，patients in the Chinese medicine group received oral Chinese medicine based on syndrome differentiation，one dose per day， twice a day. Patients in the indirect moxibustion plaster group were treated with indirect moxibustion plaster at Zusanli （ST 36）， Pishu （BL 20）， Dazhui （GV 14）， Feishu （BL 13）， Kongzui （LU 6）， and Tiantu （CV 22），once a day，40 min each time. Patients in the combination group were treated with Chinese medicine combined with indirect moxibustion plaster. Treatment lasted two weeks. Before and after treatment，the traditional Chinese medicine （TCM） symptom score，pulmonary computed tomography （CT） score，St. George's Respiratory Questionnaire （SGRQ） score，blood routine indexes ［white blood cell count （WBC），neutrophil count （NEUT），and lymphocyte count （LYM）］， and inflammatory indexes ［C-reactive protein （CRP），serum ferritin， and interleukin-6 （IL-6）］ were observed in the three groups. The clinical efficacy was evaluated. ResultAfter treatment，the scores of TCM symptoms，pulmonary CT， and SGRQ，CRP，IL-6，and ferritin in the three groups decreased（P<0.05），while WBC and LYM increased（P<0.05）， but there was no significant difference in NEUT. The above indexes in the combination group were better than those in the other two groups（P<0.05）. After treatment， the cured and markedly effective rate was 76.7% （23/30） in the combination group， 50.0% （15/30） in the Chinese medicine group， and 46.7% （14/30） in the indirect moxibustion plaster group. The cured and markedly effective rate of the combination group was significantly higher than that of the Chinese medicine group （χ²=4.593， P<0.05） and the indirect moxibustion plaster group （χ²=5.711， P<0.05）. The total effective rate was 96.7 % （29/30） in the combination group， 93.3% （28/30） in the Chinese medicine group， and 86.7% （26/30） in the indirect moxibustion plaster group. The total effective rate of the combination group was higher than that of the Chinese medicine group and the indirect moxibustion plaster group， but the differences were not statistically significant. ConclusionChinese medicine combined with indirect moxibustion plaster can effectively improve the clinical symptoms，promote pulmonary inflammation，blood routine indexes， and inflammatory indexes， and improve the quality of life of COVID-19 patients during the recovery period，which is more advantageous than Chinese medicine alone or indirect moxibustion plaster.

Objective: To investigate the effects of over expression and low expression of antisense transcripts of circular RNA cerebellar degeneration associated protein 1 (CDR1as) in Balb/C mouse bone marrow mesenchymal stem cells (BMSCs) on factors related to osteogenesis and angiogenesis. Methods: BMSCs were cultured and identified in vitro. The lentiviral (LV) vector containing the overexpressed and silenced circRNA CDR1as genes and the control lentivirus were respectively transfected into mouse BMSCs, and stable cell lines were screened. The cells were divided into the circRNACDR1as over expression group and the over expression control group, and the CircRNACDR1as low expression group and the low expression control group. The components were stained with Alizarin Red S and alkaline phosphatase after 14 and 21 days of osteoinduction; qRT-PCR was used to detect the target genes circRNA CDR1as, osteogenic differentiation markers alkaline phosphatase (ALP), runt- related transcription factor 2 (RUNX2), osteocalcin (OCN), osteopontin (OPN), osterix(Osx), collagen I (COL-1), and the mRNA expression levels of vascular endothelial grown factor (VEGF) and angiogenin-1 (Ang-1). Results: The results of alizarin red staining and alkaline phosphatase staining showed that the extracellular matrix calcium precipitation and ALP staining area of the over expression experimental group was greater than its control group, and those of the low expression experimental group was less than its control group. As the number of days of osteogenic induction increased, the calcium precipitation and ALP staining in each group also increased. RT-PCR results showed that the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in the over expression experimental group BMSCs were significantly increased (P<0.001). In the low expression experimental group, the mRNA expression levels of circRNA CDR1as, ALP, RUNX2, OCN, OPN, OSX, COL-1, VEGF and Ang-1 in BMSCs were significantly reduced (P<0.001). Conclusion Over expression of the circRNA CDR1as gene promotes the osteogenic differentiation and angiogenesis of BMSCs. Low expression of the circRNA CDR1as gene inhibits the osteogenic differentiation and angiogenesis of BMSCs.

Objective:To improve the understanding of the diagnosis and treatment of early T-cell precursor acute lymphoblastic leukemia (ETP-ALL).Methods:The clinical data of a patient with ETP-ALL who was misdiagnosed as peripheral T-cell lymphoma-not otherwise specified (PTCL-NOS) admitted to the Second Hospital of Lanzhou University in October 2020 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient who presented "inguinal lymphadenopathy" as the first symptom underwent lymph node biopsy and pathological examination at local hospital, and he was diagnosed as PTCL-NOS according to the consultation of another 2 hospitals. After 2 courses of chemotherapy (CHOPE regimen, GLD regimen, unknown specific medication and dosage), the therapeutic efficacy was poor. For further diagnosis and treatment, this patient came to Lanzhou University Second Hospital. Flow cytometry found blast cells in the bone marrow, and then other related examinations were completed, he was finally diagnosed as ETP-ALL. The chemotherapy regimens of Hyper-CVAD and EA were alternatively used, progressive disease (PD) occurred after 3 courses of treatment, and chidamide was added in the 4th and 5th courses of treatment, the disease still progressed, and the patient died after follow-up. The disease course of the patient was about 12 months.Conclusions:ETP-ALL has unique immunophenotypic characteristics. ETP-ALL patients have a low remission rate after conventional induction therapy, high recurrence rate and poor prognosis. Currently, there is no effective standard treatment regimen, and allogeneic hematopoietic stem cell transplantation or timely addition of new drugs may improve the prognosis.

Objective:To express virus-like particles of poliovirus type 2 (PV2-VLP) in insect cells using a recombinant baculovirus expressing P1 and 3CD and to preliminarily evaluate its immunogenicity.Methods:Based on the codon preference of High 5 cells, the sequences of P1 gene and 3CD gene of PV2 were optimized and inserted into pUC57-Amp to construct pUC57-PV2-P1 and pUC57-PV2-3CD. UC57-PV2-P1s mutant that carried P1 gene mutation affecting thermostability was then constructed. Recombinant baculovirus strains of rBac-PV2-P1s-3CD and rBac-PV2-P1-3CD (wild type) were constructed using homologous recombination. The expression of target proteins was detected by Western blot. PV2-VLP was purified by ion exchange chromatography. The structure of VLP was observed under transmission electron microscopy to evaluate the assembly efficiency. The immunogenicity of PV2-VLP was assessed in a rat model.Results:The recombinant baculovirus with stable expression of P1s and 3CD proteins was successfully constructed. Western blot results showed that the yield of VLP was higher after thermostability mutation than that of the wild type. A three-dimensional structure with a diameter of about 30 nm was observed under electron microscopy, indicating that the VLP was successfully assembled. Animal experiment showed that the recombinant PV2-VLP had immunogenicity and could effectively induce the production of neutralizing antibodies.Conclusions:Effective VLP vaccines could be successfully prepared using the insect cell-baculovirus expression system, which provided reference for the development of polio VLP vaccine.

Objective To study urodynamic changes of urine at different degrees of hydronephrosis based on computational fluid dynamics (CFD) method, so as evaluate the influence of hydronephrosis degree on kidneys’ ability to discharge stones. Methods Twelve models, including the branched pelvis Model A (normal hydronephrosis A1, mild hydronephrosis A2, medium hydronephrosis A3, severe hydronephrosis A4 models), mature ampullary pelvis Model B (normal hydronephrosis B1, mild hydronephrosis B2, medium hydronephrosis B3, severe hydronephrosis B4 models), and embryo pot abdominal pelvis Model C (normal hydronephrosis C1, mild hydronephrosis C2, medium hydronephrosis C3, severe hydronephrosis C4 models) were established. The urine flow velocity and velocity vector at the neck of the kidney, the outlet of the renal pelvis were calculated by CFD method. Results As the degree of hydronephrosis increased, the flow velocity of urine at the neck of the kidney and the outlet of the renal pelvis decreased. The urinary shearing force of the stones and the kidney’s ability to discharge stones gradually decreased, whereas the circulatory stagnation zone and the velocity boundary layer in kidney aggregate system gradually increased. Conclusions Hydronephrosis can cause changes in urodynamics of the urine. Therefore, the effect of hydronephrosis with different degrees on the patient’s ability to discharge stones after surgery should be fully considered， so as to choose an appropriate treatment method for kidney stones in clinic.