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Objective:To explore the effect of setting up an internal-cross disciplinary team (ICDT) in the intensive care unit (ICU) on a new model of overall treatment for patients with chronic critical illness (CCI).Methods:A 60-year-old male patient with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) admitted to ICU in the Second Affiliated Hospital of Fujian Medical University was introduced. The role of ICDT composed of physicians, nurses, respiratory therapists, physiotherapists, clinical dietitians and patients' family members in ventilator withdrawal and super-early rehabilitation was analyzed in this case.Results:The patient was diagnosed as AECOPD, type Ⅱ aspiration penumonia respiratory failure, septic shock. The ICDT in ICU carried out early rehabilitation treatment for the patient on the basis of traditional infection control and supportive treatment. Under the care of the ICDT, peripheral blood white blood cell count (WBC), neutrophil count (NEU), procalcitonin (PCT), arterial partial pressure of carbon dioxide (PaCO 2), maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP), right excursion of diaphragm, sputum viscosity, tidal volume (VT) and respiratory rate (RR) were improved. Subsequently, the ventilator mode was gradually changed and the ventilator parameters were down-regulated. The ventilator was successfully weaned on the 10th day of treatment. After weaning, the patient's bedside pulmonary function indicators improved, and he was transferred out of ICU on the 15th day of treatment and discharged on the 20th day. The mental state of the patients was good and the quality of life was greatly improved in CCI outpatient follow-up. Conclusions:ICDT cooperation is very important for monitoring and treatment of CCI patients, which is beneficial to the super-early rehabilitation and prognosis improvement of critically ill patients.
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Objective:To investigate the potential molecular mechanisms of liver cancer cell-derived secretory autophagosomes, extracellular vesicles expressing LC3B (LC3B + EVs), in promoting the exhaustion of CD8 + T cells. Methods:The proportions of LC3B + EVs and PD-1 + CD8 + T cells in peripheral blood and ascites of liver cancer patients were measured by flow cytometry. Spearman correlation test was used to analyze the correlation between the proportions of LC3B + EVs and PD-1 + CD8 + T cells. Peripheral blood mononuclear cells (PBMCs) from healthy donors were treated with LC3B + EVs or heat shock protein 90α (HSP90α) blocking antibody-pretreated LC3B + EVs for 72 h in the presence of αCD3/CD28 antibodies and IL-2 in vitro. The proportions of PD-1 + CD8 + T and IFN-γ + CD8 + T cells and the concentrations of IL-2, TNF-α and IFN-γ in the supernatants were all detected by flow cytometry. Results:The proportions of LC3B + EVs and HSP90α + LC3B + EVs in plasma and ascites from liver cancer patients were significantly higher than those in healthy control group and non-cancerous ascites group. The level of plasma LC3B + EVs, especially HSP90α + LC3B + EVs, was significantly correlated with the percentage of exhausted PD-1 + CD8 + T cells. In addition, LC3B + EVs from human liver cancer cells up-regulated the percentage of exhausted CD8 + T cells in vitro. However, LC3B + EVs pretreated with HSP90α blocking antibody could significantly inhibit LC3B + EVs-induced CD8 + T cell exhaustion. Conclusions:Liver cancer cell-derived LC3B + EVs could effectively induce CD8 + T cell exhaustion mainly through the membrane-bound HSP90α.
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ObjectiveTo investigate the effect of extracellular signal-regulated kinase 1/2 (ERK1/2) pathway inhibitor PD98059 on the proliferation and killing function of γδT cells cultured in vitro. MethodsMononuclear cells were separated from peripheral blood in healthy subjects and placed in RPMI 1640 complete medium containing zoledronic acid and interleukin-2 to obtain γδT cells through induction and culture. The ERK1/2 specific inhibitor PD98059 was used to block the ERK1/2 signaling pathway in γδT cells, and the proliferation of γδT cells was measured by cell counting, and the killing function of γδT cells was measured by CCK-8 method. Flow cytometry was used to measure the expression of granzyme B and perforin in γδT cells. The t-test was used for comparison of continuous data between groups. ResultsAfter 10 days of culture, the purity of γδT cells reached 87.94%±2.36%. The number of γδT cells treated with PD98059 was significantly lower than that of control cells ［(6.74±0.36)×105/ml vs (9.42±0.31)×105/ml, t=-12.708, P＜0.001］. Compared with the control cells, those treated with PD98059 had significantly higher positive expression rates of granzyme B and perforin (granzyme B: 48.89%±1.31% vs 41.58%±1.58%, t=7.582, P＜0.001; perforin: 65.92%±3.29% vs 33.49%±2.83%, t=15.478, P＜0001) and a significantly higher killing rate of HepG2 cells (69.28%±4.96% vs 48.34%±3.01%, t=11.201, P＜0.001). ConclusionThe ERK1/2 specific inhibitor PD98059 can inhibit the proliferation of γδT cells, but it can enhance the in vitro killing function of γδT cells.
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Objective:To investigate the effect of 2-deoxy-D-glucose (2-DG) on hunmanγδT cells on the expression of CCR5 and killing function in vitro.Methods:UsingγδT medium to cultivate peripheral blood mononuclear cell( PBMCs) in vitro.After co-cultured with various concentrations of 2-DG for 48 h,the expression of CCR5 and killing activities of γδT cells for each group were detected by flow cytometry and CCK-8 methods.Results: 2-DG could not promote the growth of γδT cells with the increase in concentration from 0 μmol/L to 1.0 μmol/L and decreased thereafter.The certain concentration ( 0-2.0 μmol/L ) of 2-DG could upregulate the expression of CCR5 in dose dependent manner.Besides,at 0.5μmol/L and 1.0μmol/L of 2-DG could increase the ex-pression of CD107a and perforin and have no effect on the granzyme B.Conclusion: Human γδT cells isolated from peripheral blood treated with 2-DG could promote the expression of CCR5 and increase the killing activities at certain concentration in vitro.
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Objective:To investigate the mechanisms of TWS119 induced CCR5 expression in hunman γδT cells. Methods:After treatment with various concentrations of TWS119 for 48h, the expression of CCR5 in γδT cells were detected by flow cytometry. The p-STAT3 and GAPDH expression were examined by Western blot analysis. Results: TWS119 could upregulate the expression of CCR5 in dose dependent manner. Western blot analysis revealed that TWS119 inhibit phosphorylation of STAT3,but had no significant impact on GAPDH. In addition, pretreatment of γδT cells with 0. 5 μmol/L STAT3 specific phosphorylation inhibitor Stattic could upregulate the expression of CCR5 and enhance the TWS119 induced CCR5 expression. Conclusion: TWS119 could upregulate CCR5 expression of γδT cells by inhibiting STAT3 phosphorylation in vitro.
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Objective To compare the clinical efficacies of electroacupuncture plus rehabilitation training versus electroacupuncture alone in treating lumbar intervertebral disc herniation and evaluate the therapeutic effect of electroacupuncture plus rehabilitation training. Method Seventy-two patients with lumbar intervertebral disc herniation were randomly allocated to a treatment group of 36 cases and a control group of 36 cases. The control group received electroacupuncture at Huatuo jiaji(Ex-B2) points. In addition to electroacupuncture, the treatment group received lumbodorsal muscle stretch training according to the outcome (forward flexion and backward extension) obtained using the Turgunmed testing and evaluating system. Isokinetic muscle strength and the activity were measured and the ODI lumbago score and the VAS pain score were recorded before and after treatment. Result There were statistically significant differences in isokinetic muscle strength and the VAS pain score (P0.05) between the treatment and control groups. Conclusion Electroacupuncture plus rehabilitation training is more effective than electroacupuncture alone in improving isokinetic muscle strength and the VAS pain score in patients with lumbar intervertebral disc herniation.
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Objective:To investigate the effect of glycogen synthase kinase-3β inhibitor TWS119 on hunman γδT cells growth and phenotypic characteristics. Methods:Using γδT medium to cultivate human peripheral blood γδT cells in vitro. After co-cultured with different concentrations of glycogen synthase kinase-3β( GSK-3β) inhibitor 4, 6-disubstituted pyrrolopyrimidine ( TWS119 ) for indicated time,growth curve and Wnt/β-catenin activation of in each group were determined by CCK-8 and Western blot assays. The CD62L and CD45RA expression theγδT cells were detected using flow cytometry. Results:Wnt/β-catenin pathway ofγδT cells could activate by TWS119. In the first group,TWS119 could upregulate the expression of CD62L and CD45RA in dose dependent manner while inhibit the growth and ratio of γδT cells. In the second group,TWS119 could promote the growth and ratio of γδT cells with the increase in concentration from 0 μmol/L to 4. 0 μmol/L and decreased thereafter. Besides,TWS119 could promote the expression of CD62L in a dose-dependent and had no effect on the CD45RA. Conclusion: Human γδT cells isolated from peripheral blood treated with TWS119 gave rise to two subsets of CD45RA+CD62L+γδT cells and CD62L+γδT cells.
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Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .
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To investigate the effects of Sangen Decoction, a compound Chinese herbal medicine, on osteoclastogenesis and bone resorption function of osteoclasts induced by polymethylmethacrylate particles in vitro.
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In China, patients with rheumatoid arthritis (RA) are often treated with traditional Chinese herbal medicine. There are certain advantages of traditional Chinese medicine therapy in treatment of RA.
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OBJECTIVE: To observe the effects of Yanghe Decoction on vascular endothelial growth factor (VEGF) in cartilage cells of osteoarthritis rabbits. METHODS: Fifteen New Zealand white rabbits were randomly divided into normal group, untreated group and Yanghe Decoction-treated group. The rabbit model of osteoarthritis was established according to Hulth's method. The rabbits were sacrificed at the 8th week after administration of Yanghe Decoction for 14 days, and then rabbit tibia articular cartilage was removed. Sections of the cartilage were stained with Safranin O for histological examination. The cartilage histological characteristics were observed according to the method of Mankin. Immunohistochemical staining was performed to investigate the expression of VEGF. Articular cartilages were observed with microscopy and image analysis method was used to measure the expression intensity of VEGF. RESULTS: There were significant differences in Mankin score between normal group and untreated group (P<0.01), and between untreated group and Yanghe Decoction-treated group (P<0.01). Immunohistochemical staining indicated that the expression intensity of VEGF in untreated group was significantly increased as compared with that in normal group (P<0.01), and also obviously higher than Yanghe Decotion-treated group. CONCLUSION: VEGF plays an important role during early stage of OA. Yanghe Decoction can protect the articular cartilage through suppressing the VEGF expression in chondrocytes and then suppress angiogenesis.
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0.05),but for patients with mild disease,active angle reproduction
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OBJECTIVE: To study the effect of melittin on apoptsis and necrosis of osteosarcoma cell line U2 OS in vitro. METHODS: Osteosarcoma cell line U2 OS was treated with melittin. The growth and proliferation was observed by MTT assay and cell counting, and the necrosis was estimated by Trypan blue staining. The cell apoptsis, Fas and Apo2. 7 expression were detected by cytometer. RESULTS: The data showed that melittin could inhibit the proliferation of U2 OS dose-dependently at 16 and 64 mg/L. Cell apoptsis was detected by cytometer, when the cells were treated by 16 mg/L and 32 mg/L of melittin respectively, and the percentages of Fas and Apo2. 7 positive cells were increased. CONCLUSION: Melittin inhibits the proliferation of osterosarcoma cell line through up-regulating Fas expression and inducing apoptsis.
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OBJECTIVE: To understand the relationship between porosity, collagen fiber orientation and strength of the plated bone after rigid plate fixation and removal. METHODS: Seventy-two New Zealand white rabbits were used in this experiment. Eight animals served as control and the other sixty-four were plated on their intact left tibiae with stainless steel (316L) 4-hole plates to induce early osteoporosis. The plates were removed 2 months after internal fixation in 40 plated animals, 8 of which were sacrificed immediately following plate removal and the other 32 were killed in successive groups with 8 in each group 1, 2, 3 and 4 months after plate removal. The remaining 24 plated animals were killed at 3, 4 and 6 months after plate fixation. After sacrifice, the samples of plated bone were prepared for light microscope, quantitative histological analysis, polarized light microscope and biomechanical test. RESULTS: The internal fixation with a rigid plate could induce the regional osteoporosis which manifested both bone loss and disorganized bone structure (loss of the orientation of the collagen fibers) leading to decreased strength of the plated bone. Although the regional osteoporosis could recover gradually after plate removal, the bone structure remained disorderly even when the bone mass returned to normal. Delayed restoration of bone structure was related to delayed restoration of bone strength. CONCLUSIONS: Besides the bone loss, the disorganized bone structure is the main cause of decrease of bone strength after rigid plate fixation and removal.
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Objective To evaluate osteogenetic effectiveness of hyaluronic acid (HA) mixed with adenovirus mediated human bone morphogenetic protein-2 gene(Adv-hBMP-2) transfected bone marrow derived mesenchymal stem cells(BMSCs) of rabbits in the repair of radial shaft defects of rabbits. Methods The BMSCs were collected from bone marrow of rabbits, cultured and transfected by Adv-hBMP-2 in vitro. The radial shaft defects (1.5 cm) models of 20 rabbits (40 sides) were created and then respectively injected the cells and/or materials according to the following four groups (n=10): Ⅰ, Adv-hBMP-2 transfected BMSCs+HA; Ⅱ, Adv-hBMP-2 transfected BMSCs; Ⅲ, HA; Ⅳ, control. Results The bone defects of 6 out of 8 sides in the group Ⅰ were repaired completely and partial bone marrow cavities were revasculized. Only 3 sides were repaired completely in the group Ⅱ and no side was repaired completely in the groups Ⅲ and control. There were statistic significance between each groups in X-ray scores. In the groups ofⅠand Ⅱ, much woven callus formed in the sites of the bone defects in the 4-8th week after injection. Partial bone marrow cavities were revasculized in the 12th week after injection; while in the groups of HA and control, the sites of bone defects were mainly full of fibrous tissue and no healing happened. Both ends of the bone defects were hardened. There was significant difference statistically in the new trabecular bone area in the groups ofⅠand Ⅱ. The maximum vertical loading and elastic modulus were significant differenct in the group ⅠandⅡ. The ratios of average maximum vertical loading/normal value of radial shaft in the groups ofⅠandⅡwere 75.86% and 45.45%, respectively. Conclusion The tissue-engineered bone made by HA mixed with Adv-hBMP-2 transfected BMSCs can repair the bone defects in the radial shaft of rabbits. HA is a safe, effective carrier for tissue engineering.