Usage of mobile health interventions among overweight/obese PCOS patients undergoing assisted reproductive technology treatment during the COVID-19 pandemic.

Objective: In the context of the coronavirus disease 2019 (COVID-19) pandemic, telemedicine is a promising tool for providing clinical care for patients. Since the first-line treatment for infertile women with polycystic ovarian syndrome (PCOS) is lifestyle modification, a mobile-based service that provides lifestyle modification education would be helpful in the treatment of PCOS patients. In this observational study, the effect of a mobile Health (mHealth) application for lifestyle modification on PCOS patients undergoing assisted reproductive technology (ART) treatment was evaluated.Methods: A total of 79 overweight/obese patients (40 in the paper group and 39 in the WeChat application group) with PCOS from the First Affiliated Hospital of University of Science and Technology of China were enrolled in the study. The changes in the outcomes of BMI and ART treatment were analyzed between the two groups.Results: After three months of intervention, the BMIs in the control and mHealth groups were 24.5 ± 3.3 and 23.7 ± 3.1, respectively. The percentage of patients who lost weight was higher in the WeChat group than in the control group (87.2% vs. 67.5%). Furthermore, PCOS patients in the WeChat group were found to have a higher live birth rate than those in the control group (p = 0.005).Conclusion: Lifestyle modifications for PCOS patients undergoing ART treatment using the WeChat application improved weight loss and oocyte quality. Infertile patients with PCOS were more likely to make lifestyle modifications based on the usage of mobile applications during the COVID-19 pandemic.

Real-Time Analysis of the Stability of Oil-In-Water Pickering Emulsion by Electrochemical Impedance Spectroscopy.

In this paper, electrical impedance spectroscopy (EIS) was applied to investigate the stability of oil-in-water (O/W) Pickering emulsions prepared with negatively charged silica nanoparticles in combination with a trace amount of redox switchable fluorescent molecules, ferrocene azine (FcA). Electrical impedance values of emulsions obtained at different emulsification speeds were estimated according to the frequency response data with frequencies ranging from 1 MHz to 1 Hz. The equivalent circuit model of toluene-in-water emulsion was established by the resistor (RO/W) and capacitor (CO/W) in parallel connection. Nyquist diagrams for the emulsions prepared by toluene and water were characterized by the formation of one semi-circle. The droplet size distribution is one of the important factors that affect the stability of the emulsion, except for the volume fraction of water and oil, the size of stabilizing particles, etc. The average particle size of the emulsion droplets decreased as the emulsification speed increased, indicating the higher stability of the emulsion. It was found that the fitted impedance value RO/W of the emulsion decreased with decreasing particle size prepared at different emulsification speeds and storage time by performing real-time EIS detection techniques. The results suggested that EIS could be used to characterize the stability of a toluene-in-water emulsion stabilized by FcA modified silica nanoparticles. Moreover, based on the good electrochemical activity of the FcA molecule, the stability of the Pickering emulsion can be modulated by adding oxidant and reductant and detected by EIS in real-time.

Redox-Responsive Pickering Emulsions Based on Silica Nanoparticles and Electrochemical Active Fluorescent Molecules.

In this paper, we report a novel redox-responsive water-in-oil Pickering emulsion stabilized by negatively charged silica nanoparticles in combination with a trace amount of redox switchable fluorescent molecule ferrocene azine (FcA), in which ferrocene serves as a redox-sensitive group and anthryl unit serves as a fluorescence emission center. By alternately adding oxidants and reducing agents at a moderate condition, the amphiphilicity of silica nanoparticles changes because of the adsorption of Fc+A and the desorption of FcA on the silica surface. On the one hand, the stability of emulsions can be transformed between stable and unstable at ambient temperature via redox trigger and the regulation process can be cycled at least three times. On the other hand, the fluorescent intensity of the FcA molecule can be regulated by redox stimuli; thus, the change in fluorescent behavior of the emulsion droplets is observed upon redox cycles, which makes it useful in the fluorescent label of stimuli-responsive Pickering emulsions. This work provides a deep understanding of the regulation mechanism of Pickering emulsions upon redox stimuli and opens the new way for in situ fluorescent label of stimulus-responsive Pickering emulsions without introducing additional fluorescent molecules.

Preparation of Core-Shell Silver Nanoparticles@Mesoporous Silica Nanospheres with Catalytic Activities.

Core-shell silver nanoparticles@mesoporous silica spherical nanoparticles (Ag NPs@MSNs) were prepared by a two-step method. First, Ag NPs were synthesized by chemical reduction using silver nitrate (AgNO3) as the precursor, cetyl trimethyl ammonium bromide (CTAB) as the stabilizer, and sodium borohydride (NaBH4) as the reductant. Then, MSNs were obtained by employing CTABstabilized Ag NPs as the template and hydrolyzing tetraethoxysilane (TEOS) precursor in the presence of the alkaline precipitant, triethanolamine (TEOA). The effects of different preparation routes (core-first vs. shell-first), type of reductants as well as extraction methods and agents were studied. The obtained core-shell NPs were characterized by infrared spectroscopy (IR) and transmission electron microscopy (TEM). Our results showed that the core-first route was viable to produce uniform Ag NPs@MSNs with ordered mesostructures. Afterwards, those NPs were used as the catalyst to catalyze the reduction of rhodamine, a model dye compound representing organic pollutants in waste water, in the presence of NaBH4. It was found that Ag NPs@MSNs not only were efficient catalysts but also participated as coreductants in the reaction. Moreover, they exhibited almost no loss of catalytic efficacy after several reduction cycles, which indicated their promising future use as efficient recyclable catalysts for organic pollutant treatments.

Thermally Stable Metallic Nanoparticles Prepared via Core-Cross-linked Block Copolymer Micellar Nanoreactors.

Thermally stable metallic nanoparticles (MNPs) are highly desirable for the melt processing of polymer nanocomposites. However, due to the high surface energy penalty and decreased melting temperature, MNPs are easy to agglomerate and lose their unique properties if there is no protection or confinement layer. In this work, we report a facile and efficient way to synthesize thermally stable MNPs using core-cross-linked polystyrene-b-poly(4-vinylpyridine) (PS-b-P4VP) reverse micelles as nanoreactors. From infrared results, gold, silver, and palladium ions exhibited distinctive coordination to the 4VP groups with varying chelation strengths. Compared to the non-cross-linked micelles, 1,4-dibromobutane (DBB)-cross-linking of the P4VP cores provided several advantages. First, it prevented severe swelling of the P4VP cores caused by the reducing agents and subsequent merger of swollen micelles. Second, the quaternized P4VP with hydrophilicity enhanced the uptake speed of precursor metal ions into the cores. Third, the cross-linked cores greatly stabilized the MNPs against the high-temperature environment (e.g., 110 °C for 40 h in toluene). In addition, the solubility of the reducing agents also played an important role. Anhydrous hydrazine could swell the P4VP cores and concentric core-shell particle morphology was obtained. On the contrary, triethylsilane could not swell the P4VP cores and thus eccentric core-shell particle morphology was observed. Only the concentric core-shell MNPs exhibited good thermal stability, whereas the eccentric core-shell MNPs did not. This work suggested that these thermally stable MNPs could be good candidates for the melt processing of functional polymer nanocomposites.

Fentanyl inhibits the invasion and migration of colorectal cancer cells via inhibiting the negative regulation of Ets-1 on BANCR.

BACKGROUND: Recent studies have shown the potential anti-tumor effect of fentanyl on colorectal cancer (CRC). However, its underling mechanism is still unclear. Since studies indicates the abnormal expression of transcription factor Ets-1 and BRAF-activated lncRNA (BANCR) in CRC progress, the relationship between Ets-1 and BANCR was investigated here to illustrate the fentanyl-induced mechanism on CRC in vitro. METHODS: The expression levels of Ets-1 and BANCR were first detected in fentanyl-treated CRC cells. The interaction between Ets-1 and BANCR promoter was verified with chromatin immunoprecipitation assays, as well as corresponding acetylation of histones. The regulation of Ets-1 on BANCR expression was confirmed through luciferase assays and RT-PCR analysis. And, cell clone formation, cell migration and invasion were observed to evaluate the anti-tumor effects of fentanyl. Ets-1 overexpression or co-overexpression with BANCR was further performed by plasmids transfection to show the regulatory role of Ets-1 in fentanyl-induced mechanism. RESULTS: Fentanyl induced BANCR upregulation and Ets-1 downregulation in CRC cells. Further studies showed that Ets-1 negatively regulated BANCR expression via the deacetylation of histones H3 within BANCR promoter. Moreover, fentanyl induced less cell clone formation, as well as inhibited cell migration and invasion in vitro, while Ets-1 overexpression inhibited fentanyl-induced effects that could be reversed by BANCR co-overexpression. CONCLUSION: Fentanyl showed anti-tumor like effects on CRC cells, including less cell clone formation and inhibited cell migration and invasion. Furthermore, the regulatory role of Ets-1 on BANCR influenced fentanyl-induced mechanism, indicating their potential application in the therapeutic treatment of CRC.

Renshen Yangrong decoction for secondary malaise and fatigue: network pharmacology and Mendelian randomization study.

Background: Renshen Yangrong decoction (RSYRD) has been shown therapeutic effects on secondary malaise and fatigue (SMF). However, to date, its bioactive ingredients and potential targets remain unclear. Purpose: The purpose of this study is to assess the potential ingredients and targets of RSYRD on SMF through a comprehensive strategy integrating network pharmacology, Mendelian randomization as well as molecular docking verification. Methods: Search for potential active ingredients and corresponding protein targets of RSYRD on TCMSP and BATMAN-TCM for network pharmacology analysis. Mendelian randomization (MR) was performed to find therapeutic targets for SMF. The eQTLGen Consortium (sample sizes: 31,684) provided data on cis-expression quantitative trait loci (cis-eQTL, exposure). The summary data on SMF (outcome) from genome-wide association studies (GWAS) were gathered from the MRC-IEU Consortium (sample sizes: 463,010). We built a target interaction network between the probable active ingredient targets of RSYRD and the therapeutic targets of SMF. We next used drug prediction and molecular docking to confirm the therapeutic value of the therapeutic targets. Results: In RSYRD, network pharmacology investigations revealed 193 possible active compounds and 234 associated protein targets. The genetically predicted amounts of 176 proteins were related to SMF risk in the MR analysis. Thirty-seven overlapping targets for RSYRD in treating SMF, among which six (NOS3, GAA, IMPA1, P4HTM, RB1, and SLC16A1) were prioritized with the most convincing evidence. Finally, the 14 active ingredients of RSYRD were identified as potential drug molecules. The strong affinity between active components and putative protein targets was established by molecular docking. Conclusion: This study revealed several active components and possible RSYRD protein targets for the therapy of SMF and provided novel insights into the feasibility of using Mendelian randomization for causal inference between Chinese medical formula and disease.

Design of Proanthocyanidins and TiO2 Nanoparticles-Based Novel Emulsions as a Platform for UV Protection.

A white oil-in-water novel emulsion stabilized by TiO2 nanoparticles with UVB shielding properties and proanthocyanidins with antioxidant activity was prepared, where the proanthocyanidins aggregated at the oil-water interface to reduce interfacial tension while TiO2 nanoparticles were dispersed in the continuous water phase to hinder droplet coalescence. It was found that the average oil droplet size was less than 10 µm and decreased with the increase of proanthocyanidins concentration, but the increase of the content of TiO2 nanoparticles had little effect on it. The combination of TiO2 nanoparticles and proanthocyanidins was versatile for oil phases with different polarities, and the resulting emulsion exhibited high stability in the face of centrifugation, heating and prolonging storage time. After encapsulating the UVA filter avobenzone in white oil, the emulsion was endowed with the ability to resist UVB and UVA. Further, the emulsion showed great free radical scavenging ability for superoxide anion radical (⋅O2 - ), hydroxyl radical (⋅OH) with the clearance rate of over 70 %, indicating the good antioxidant activity. The ingenious combination of UVB, UVA filter and antioxidant with emulsion as carrier provides a new idea for the preparation of full-band sunscreen emulsion.

Improvements of Solubility and Bioavailability of Lutein Through Grafting with Hydrophilic Polyacrylic Acid.

In this study, polyacrylic acid grafted lutein (PAA-g-lutein) was prepared by hydrophilic modification of lutein with polyacrylic acid (PAA) through Steglish esterification method. The unreacted lutein was loaded in micelles formed by self-assembly of graft copolymers in water to form composite nanoparticles. The bioaccessibility and bioavailability of lutein nanoparticles were studied by in vitro and in vivo digestion experiments. Compared with free lutein, the saturated solubility and bioaccessibility of lutein nanoparticles were increased by 78 times and 3.6 times, respectively. The pharmacokinetics results in the mice model showed that the maximum concentration (Cmax) and area under concentration-time curve (AUC) of plasma of mice were increased by 3.05 and 6.07 times with lutein nanoparticles compared with free lutein. Meanwhile, the prepared lutein nanoparticles also promoted the accumulation of lutein in the liver, mesenteric adipose, and eyeballs. These results indicate that graft copolymerization of lutein with water-soluble polymers to form nanoparticles is an effective method to promote the bioavailability of lutein in vivo. Moreover, this method is simple and applicable, and can also be used for the modification of other bioactive molecules.

A prognostic model of drug tolerant persister-related genes in lung adenocarcinoma based on single cell and bulk RNA sequencing data.

Background: Acquired resistance to targeted drugs is a major challenge in cancer. The drug-tolerant state has been proposed to be an initial step towards acquisition of real drug-resistance. Drug tolerant persister (DTP) cells are purported to survive during treatment and stay dormant for several years. Single cell sequencing can provide a comprehensive landscape of gene expression in DTP cells, which can facilitate investigation of heterogeneity of a drug tolerant state and identification of new anticancer targets. Methods: The genetic profiling of DTPs was explored by integrating Gene Expression Omnibus (GEO) datasets, and a prognostic signature of DTP-related genes (DTPRGs) in lung adenocarcinoma of TCGA LUAD cohort was constructed. The scores of infiltrating immune cells were calculated and activity of immune-related pathways was evaluated by single-sample gene set enrichment analysis (ssGSEA). Functional enrichment analysis of the DTPRGs between low- and high-risk groups was performed. Immune cell subtypes and immune-related pathways were analyzed. Results: An 11-gene panel (MT2A, UBE2S, CLTB, KRT7, IGFBP3, CTSH, NPC2, HMGA1, HNRNPAB, DTYMK, and IHNA) was established. DTPRGs were mainly correlated with nuclear division, chromosome segregation, and cell cycle pathways. Infiltration of immune cells was lower in the high-risk group while the inflammation-promoting and MCH-class I response pathway had higher activity in the high-risk group. A nomogram was generated with prognostic accuracy, further validated using clinical outcomes following therapy with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs). Discussion: A prognostic model of lung adenocarcinoma based on DTPRGs was constructed. Targeting DTP cells is a potential therapeutic approach to prevent a drug tolerant state.

Propofol exerts neuroprotective functions by down-regulating microRNA-19a in glutamic acid-induced PC12 cells.

BACKGROUND: Propofol, a kind of intravenous sedative drug, is certified that exerts anti-inflammation and antitumor functions. However, the influence of propofol in cerebral injury and the corresponding mechanism remains unexplained, that our article focuses on. METHODS: PC12 cells were treated with propofol and exposed in glutamic acid (Glu) solutions. Cell viability, apoptotic potential, apoptosis-related and autophagy-linked proteins were tested via CCK-8, flow cytometry, and western blot assays. Reverse transcription-quantitative real-time PCR was utilized to test miR-19a expression in Glu-stimulated cells. Next, miR-19a mimic transfection was used to assess the effects of miR-19a on cell apoptosis and autophagy in Glu or propofol treated cells. Finally, western blot was performed to test AMPK and mTOR pathways. RESULTS: Glu exposure promoted cell apoptosis and autophagy of PC12 cells, while propofol attenuated cell apoptosis and autophagy triggered by Glu. Additionally, propofol decreased the miR-19a expression in Glu-stimulated PC12 cells. Meanwhile, over-expression of miR-19a reversed the effects of propofol on Glu-induced cell apoptosis and autophagy. Moreover, propofol potentiated AMPK and mTOR pathways in Glu-stimulated PC12 cells via impeding miR-19a expression. CONCLUSIONS: These finding revealed that propofol relieved Glu-triggered apoptosis and autophagy of PC12, and activated AMPK and mTOR pathways by suppressing miR-19a expression.

The efficacy of combined adductor canal block with local infiltration analgesia for pain control after total knee arthroplasty: A meta-analysis.

BACKGROUND: This meta-analysis aimed to evaluate the efficiency and safety of the combined adductor canal block (ACB) with local infiltration anesthesia (LIA) versus LIA alone for pain control after total knee arthroplasty (TKA). METHODS: We searched PubMed, Medline, Embase, Web of Science, the Cochrane Library and Google databases from inception to August 2017 to selected studies that comparing the combined ACB with LIA and LIA alone for pain control after TKA. Only randomized controlled trials (RCTs) were included. Outcomes included visual analogue scale (VAS) with rest or mobilization at 8 h, 24 h and 48 h, total morphine consumption at 6 h, 24 h and 48 h, distance walked at 24 h and 48 h and the length of hospital stay. RESULTS: Seven randomized controlled trial (RCTs) were finally included in this meta-analysis. The present meta-analysis indicated that, compared with LIA alone, combined ACB with LIA was associated with a reduction of VAS with rest at 24 h and 48 h and VAS with mobilization at 24 h. Additionally, combined ACB with LIA was associated with an increase of the distance walked at 24 h and a reduction of the length of hospital stay. CONCLUSION: Combined ACB with LIA could significantly reduce pain scores and morphine consumption compared LIA alone after TKA. Further multimodal large sample RCTs are needed to identify the optimal drug of ACB and LIA.

Development of bupivacaine decorated reduced graphene oxide and its local anesthetic effect-In vivo study.

The present works aims to develop bupivacaine modified reduced graphene oxide (BPV/RGO), and comparative evaluation of their anesthetic effect with free bupivacaine (BPV). The prepared BPV/RGO was studied by using various spectroscopic and microscopic characterization studies. In vitro drug release from BPV/RGO was studied using HPLC analysis. The cytotoxicity of BPV/RGO was studied against fibroblast (3T3) cells. In vivo evaluation of anesthetic effects was performed on animal models. BPV/RGO showed a prolonged in vitro release and lower cytotoxicity when compared to free BPV. Also, BPV/RGO showed a significantly prolonged analgesic effect when compared to free BPV. Further, the prepared BPV/RGO drug delivery system demonstrated to function as gifted to overcome the drawbacks of free BPV and other available drug delivery systems by prolonging the anesthetic effect with poor cytotoxicity.

Evaluation of local anesthetic effects of Lidocaine-Ibuprofen ionic liquid stabilized silver nanoparticles in Male Swiss mice.

A simple approach for the synthesis of Lidocaine-Ibuprofen ionic liquid stabilized silver nanoparticles (IL-AgNPs) was reported in this work. The shape, size and surface morphology of the Lidocaine-Ibuprofen ionic liquid stabilized AgNPs were characterized by using spectroscopic and microscopic techniques such as Ultraviolet-visible spectroscopy (UV-Visible), X-ray diffraction (XRD) analysis, Selected area electron diffraction (SAED), Transmission electron microscopy (TEM). TEM analysis showed the formation of 20-30nm size of IL-AgNPs with very clear lattice fringes. SAED pattern confirmed the highly crystalline nature of fabricated IL stabilized AgNPs. EDS results confirmed the formation of nanosilver. The fabricated IL-AgNPs were studied for their local anesthetic effect in rats. The results of local anesthetic effect showed that the time for onset of action by IL-AgNPs is 10min, which is significantly higher than that for EMLA. Further, tactile test results confirmed the stronger and faster local anesthetic effect of IL-AgNPs when compared to that of EMLA.

The efficiency of intravenous acetaminophen for pain control following total knee and hip arthroplasty: A systematic review and meta-analysis.

BACKGROUND: This meta-analysis aimed to evaluate the efficiency and safety of intravenous acetaminophen as an adjunct to multimodal analgesia for pain control after total joint arthroplasty (TJA). METHODS: PubMed, Embase, Web of science, Medline, and Cochrane library databases were systematically searched. Randomized controlled trials (RCTs) and non-RCTs were included. Fixed/random effect model was used according to the heterogeneity tested by I statistic. Meta-analysis was performed using Stata 11.0 software. RESULTS: Four studies including 865 patients met the inclusion criteria. The present meta-analysis indicated that there were significant differences between groups in terms of pain scores at 24 hours (weighted mean difference [WMD] = -0.926, 95% confidence interval [CI]: -1.171 to -0.681, P = .000), 48 hours (WMD = -0.905, 95% CI: -1.198 to -0.612, P = .000), and 72 hours (WMD = -0.279, 95% CI: -0.538 to -0.021, P = .034). Significant differences were found regarding opioid consumption at 24 hours (WMD = -4.043, 95% CI: -5.041 to -3.046, P = .000), 48 hours (WMD = -5.665, 95% CI: -7.383 to -3.947, P = .000), and 72 hours (WMD = -6.338, 95% CI: -7.477 to -5.199, P = .000). CONCLUSION: Intravenous acetaminophen was efficacious for reducing postoperative pain and opioid consumption than the placebo following total joint arthroplasty. Due to the limited quality of the evidence currently available, more RCTs are needed.

Preoperative intravenous glucocorticoids can decrease acute pain and postoperative nausea and vomiting after total hip arthroplasty: A PRISMA-compliant meta-analysis.

BACKGROUND: A systematic review and meta-analysis of published randomized controlled trials (RCTs) were performed to assess the efficacy and safety of preoperative intravenous glucocorticoids versus controls for the prevention of postoperative acute pain and postoperative nausea and vomiting (PONV) after primary total hip arthroplasty (THA). METHODS: A computer literature search of electronic databases, including PubMed, Embase, the Cochrane Central Register of Controlled Trials (CENTRAL), Web of Science, China National Knowledge Infrastructure (CNKI), and China Wanfang database, was conducted to identify the relevant RCTs comparing preoperative intravenous glucocorticoids versus placebos for reducing acute pain and PONV in THA patients. The primary outcomes included the use of the visual analog scale (VAS) with rest or mobilization at 6, 24, 48, and 72 hours and the occurrence of PONV. The secondary outcome was total morphine consumption. We calculated the risk ratio (RR) with a 95% confidence interval (95% CI) for dichotomous outcomes, and the weighted mean difference (WMD) with a 95% CI for continuous outcomes. RESULTS: Pooled data from 7 RCTs (411 THAs) favored preoperative intravenous glucocorticoids against acute pain intensity at 4, 24, and 48 hours (P < .05). There was no significant difference between the VAS with rest or mobilization at 72 hours (P > .05). Subsequently, preoperative intravenous glucocorticoids provided a total morphine-sparing effect of 9.36 mg (WMD = -9.36, 95% CI = -12.33 to -6.38, P = .000). In addition, preoperative intravenous glucocorticoids were associated with a significant reduction of the occurrence of PONV (RR = 0.41, 95% CI = 0.30-0.57, P = .000). CONCLUSION: Intravenous glucocorticoids can decrease early pain intensity and PONV after THA. However, the low number of studies and variation in dosing regimens limits the evidence for its use. Thus, more high-quality RCTs are still needed to identify the optimal drug and the safety of intravenous glucocorticoids.

Study on blocking the leukemia immune escape after BMT by Fas-Fas ligand pathway.

BACKGROUND: To investigate if bone marrow transplantation (BMT) with bone marrow mononuclear cells (BMMCs) transducted with murine soluble Fas gene (sFas) using adenovirus vector could block the immune escape of leukemia cells eliminate the residual leukemia cells and reduce their relapse. METHODS: The recombinant adenovirus vector with murine sFas, adsFas, and the control vector adEGFP were constructed using homologous recombination between two plasmids in Escherichia coli. BMT was carried out after the BMMCs were infected with Adenoviruses. The mice models of leukemia/lymphoma were constructed by inoculating female C57BL/6 mice (H-2b) with 10(5) EL4 cells/mouse through caudal vein. Donors of bone marrow grafts were syngeneic male mice. BMMCs were infected with AdsFas or AdEGFP 24 hours before (Group D or E). The following three groups were simultaneously used: Group A, no BMMCs transplanted; Group B, transplanted with BMMCs not infected with adenoviruses; Group C, only transfusing EL4 cells, neither irradiation nor BMT. The hematopoietic reconstitution, generation of leukemia/lymphoma and the survival rate were observed in all groups after BMT. RESULTS: The adenovirus vectors were successfully constructed. The titre of virus after purification was up to 2.5 x 10(11) pfu/ml. Spleen indices examined 11 days after BMT were not obviously different among Group B, D and E (P > 0.05), but indices in Group A were significantly lower than those in the latter three groups (P < 0.01). Counts of leukocytes and platelets on +30 day showed mice were reconstituted satisfactorily in Group B and D, but very low in Group C and E. The Y-chromosomes existed 2 months after BMT and examination of bone marrow cytology showed that Group B and D were almost normal, but Group C and E had plenty of lymphoblast-like tumor cells. Tumors were obviously observed in the mice of Group C and E by histopathological examination, but the mice in Group B and D were normal. The survival rates were 0 (0/4) in Group A, 100% in Group B (6/6) and D (16/16), 12.5% (2/16) in Group C and 6.25% (1/16) in Group E respectively. It is demonstrated that, in contrast with the control (Group EGFP), survival rate was significantly increased in the sFas Group (P < 0.01). CONCLUSIONS: The transfer of sFas gene by adenovirus changed the prognosis state of leukemia/lymphoma mice after auto-BMT. The transduction of sFas might block the effect of the immune escape of EL4 cells through FasL. These results could thus provide a new direction to find a way to treat the leukemia and its recurrence after BMT.

Cloning and expression of the cDNA of a murine soluble Fas.

In order to regulate the apoptosis induced by Fas-FasL system, a soluble isoform of mouse Fas was cloned from thymocytes of immature mice with the primers designed according to the full-length Fas cDNA sequence in the GeneBank. It was directionally inserted into the intermedium vector pUC19. DNA sequencing proved that it was consistent with the expected sequence. Then it was subcloned into the eukaryotic expression vector pCA13, which was used to construct the recombinant vector pCA13-FasC. By lipofectamine (LF2000)-mediated transfection, pCA13-FasC was transfected into the 293 cells. RT-PCR and Western blot indicated that the murine soluble Fas C protein was expressed in the 293 cells. Apoptosis inducing test showed that the expression of this murine Fas C could block the Fas-induced apoptosis, which confirmed the biological activity of the recombinant Fas C.

[FasL-cDNA transfected into mouse bone marrow cells ex vivo to prevent graft versus host disease].

To explore the new approach to prevent graft versus host disease (GVHD) by purging ex vivo T lymphocytes of bone marrow graft through Fas-FasL way, FasL-cDNA was transfected into BALB/c mouse bon e marrow cells by liposome ex vivo. The transfected cells were cultured together with BAC (BALB/c x C57BL/6) mouse bone marrow graft. The mixing bone marrow graft was infused into BALB/c mouse recipients after 60Co-gamma irradiation. The mortality, manifestation and pathologic change of GVHD in recipient mice were observed. The CFU-S and Y chromosome from donor mice were detected. The results showed that compared with control group, the mortality in 60 days of the recipients in the experimental group decreased (20% vs 70%, P < 0.01) and the morbidity of GVHD lowered (40% vs 100%, P < 0.01). The CFU-S counts for all groups were at normal level on 20 days after transplantation. The Y chromosome from donor mice was discovered in 70% bone marrow nucleated cells of recipient mice survived over 2 months in the experimental group. It is concluded that mFasL-cDNA transfected mouse bone marrow cells prevent GVHD after culturing together with bone marrow graft, and accelerate hematopoietic reconstitution in recipient mice.

[Study on expression and function of fas ligand in human myeloid leukemia cells].

To determine whether the Fas receptor-Fas ligand (FasR-FasL) system, which triggers apoptosis in sensitive cells, is an important mechanism of cytotoxicity in myeloid leukemia. FasL expression was investigated in myeloid leukemia cells and its upregulation by a combination of IL-2 and INF-gamma, +/- as well as its function of inducing Jurkat cells to apoptosis mainly by flow cytometry (FCM). Results showed that leukemia cells expressed more FasL (3.59 +/- 1.05)% than that expressed in the healthy individuals (0.36% +/- 0.16)%, P < 0.001 and the FasL was upregulated (7.78 +/- 3.40)%, P < 0.01 when treated with IL-2 and IFN-gamma. Leukemia cells were co-cultivated with Jurkat cells for 24 hours. Then Jurkat cells were labeled with FITC-annexin V and PE-CD3 to assess apoptosis by FCM. The leukemia cells, which had been incubated with IL-2 and IFN-gamma, induced more Jurkat cells to apoptosis than the ones that freshly isolated from the peripheral blood mononuclear cells, which raised the figure from (8.28 +/- 1.61)% to (10.73 +/- 2.16%). And the supernatant derived from the former killed more Jurkat cells than the latter. It was concluded that human myeloid leukemia cells expressed high levels of functional FasL that can kill Jurkat T-cells by apoptosis. FasR-FasL sys tem could play a role in the "immune escape" and relapse of the leukemia. The induction of apoptosis through the Fas pathway might be a novel and effective approach for leukemia immunotherapy.