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Sempervirine, a yohimbane-type alkaloid isolated from Gelsemium elegans, was found to significantly inhibit the cellular proliferation of U251 cells in vitro and in vivo in a dose-dependent manner. U251 cells were treated with 0-16 μmol·L-1 of sempervirine for 24, 48 or 72 h. An MTT assay and clone formation assay were used to investigate cell survival and clone formation. Hoechst staining and Annexin V-FITC/PI staining were used to measure cell apoptosis. The expression of PI3K, AKT, p-AKT, Bax, Bcl-2, caspase-3 and cleaved caspase-3 was determined by Western blot analysis. The antitumor effect of sempervirine in vivo was investigated by inoculating nude mice with U251 cells. All animal experiments were in strict accordance with the regulations of the Biomedical Ethics Committee of Fujian Medical University (Fujian, China). The results show that sempervirine significantly inhibits the proliferation and induces the apoptosis of U251 cells, promotes cleavage of caspase-3, down-regulates the protein expression of PI3K and Bcl-2/Bax, and inhibits phosphorylation of AKT in vitro. Intraperitoneal injection of 4 or 8 mg·kg-1·day-1 of sempervirine inhibits U251 cells tumor growth in the xenograft nude mice, and tumor weight decreased by 44.76% and 61.26%, respectively. Our study shows that sempervirine significantly inhibits the proliferation of U251 cells in vitro and in vivo, laying a foundation for further research and development of its anti-glioma effect.
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OBJECTIVE@#To summarize and critically assess the inhibitory effects of Chinese herbal medicine (CHM) on tumor volume and tumor weight for the treatment of osteosarcoma (OS) in mouse models.@*METHODS@#PubMed, Embase, Web of Science, China Knowledge Resource Integrated Database (CNKI), Wanfang Database, VIP Database, and Chinese BioMedical (CBM) were searched since their inception dates to March 10, 2016. Two reviewers independently selected the controlled studies estimating effects of CHM on mouse OS by administration in vivo. A pair-wise meta-analysis was performed. Twenty-five studies with adequate randomization were included in the systematic review.@*RESULTS@#CHM may significantly inhibit OS growth in mice, as assessed using the tumor weight [20 studies, n=443; 290 for CHM and 153 for the control: pooled mean difference (MD)=-2.90; 95% confidence interval (Cl): -3.50 to -2.31: P<0.01], tumor volume (16 studies, n=382; 257 for CHM and 125 for the control; pooled MD =-2.57; 95% Cl: -3.33 to -1.80; P<0.01) and tumor growth inhibition rate.@*CONCLUSION@#CHM could significantly inhibit the growth of OS in mouse models, which might be supportive for the design of preclinical and clinical trials in future.
Subject(s)
Animals , Mice , Drugs, Chinese Herbal , Therapeutic Uses , Osteosarcoma , Drug Therapy , Publication Bias , Risk Factors , Tumor Burden , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE@#To observe the clinical therapeutic effect of three-needle therapy combined with acupuncture on complex regional pain syndrome typeⅠ(CRPS-Ⅰ) after stroke.@*METHODS@#A total of 96 patients with CRPS-Ⅰ after stroke were randomized into an observation group and a control group, 48 cases in each one. In the control group, based on the routine treatment, acupuncture was applied at Neiguan (PC 6), Shuigou (GV 26), Sanyinjiao (SP 6) and other supplementary acupoints, once a day, 6 times a week for 3 weeks. Based on the treatment in the control group, three-needle therapy was added at coracoid process, greater tuberosity of humerus and infraglenoid tubercle, the muscular fascia was released by fan-shaped separation technique for 3-6 times, and the treatment was given once every 2 days, 3 times a week for 3 weeks. Visual analogue scale (VAS) score, hand swelling level, Fugl-Meyer assessment (FMA) score and Barthel index score were observed to evaluate the pain and swelling severity, motor function and living ability of patients before and after treatment.@*RESULTS@#① Compared before treatment, the VAS score, hand swelling level, the FMA score and the Barthel index after treatment and 40 days after treatment were improved in both of the two groups (0.05).@*CONCLUSION@# three-needle therapy combined with acupuncture can improve the pain severity and the motor function of affected limbs in patients with CRPS-Ⅰ after stroke, and the therapeutic effect may be sustained for a long term. However, the treatment seems to be ineffectual on extremity swelling.
Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , Complex Regional Pain Syndromes , Therapeutics , Needles , Stroke , Treatment OutcomeABSTRACT
objective To investigate the effect of toll-like receptor 4 (TLR4) in RAW264.7 macrophage inflammation induced by palmitic acid and its possible mechanisms thereof. Methods C57BL/6J male mice were fed normal (control group) or high-fat diet (high-fat group) for 18 weeks. The serum level of free fatty acids (FFA) and mRNA levels of inflammatory cytokines in visceral adipose (TNF-α, IL-6 and MCP-1) were detected. Then, palmitic acid (150μmol/L and 300 μmol/L) was applied to stimulate RAW264.7 macrophage, the mRNA expression and the secretion level of cytokines were detected. Meanwhile, TLR4 protein level, the nuclear translocation and intracellular level of NF-κB p65 were also detected. After TLR4 inhibition by siRNA transfection, the mRNA expression and the secretion level of cytokines were detected. Results The body weight and Lee's index were significantly higher in high-fat group than those of control group (P<0.05). The serum level of FFA and mRNA levels of inflammatory cytokines (TNF-α, IL-6 and MCP-1) in visceral adipose were significantly higher in high-fat group than those of control group (P<0.05). Compared with mice without FFA (BSA group), the mRNA expression and the levels of TNF-α, IL-6, and MCP-1 were significantly increased (P<0.05), and the protein expression of TLR4 and NF-κB phosphorylated protein were up-regulated (P<0.05) in mice treated with palmitic acid (150μmol/L and 300μmol/L). The nuclear translocation and intracellular level of NF-κB p65 were increased in mice treated with palmitic acid 300μmol/L than mice in BSA group (P<0.05). The mRNA levels of TNF-α, IL-6 and MCP-1 and their secretion level were significantly decreased after TLR4 siRNA transfection (P<0.05). Conclusion These results suggest that TLR4 may mediate the inflammation in RAW264.7 macrophage stimulated by palmitic acid by inducing inflammatory cytokines (TNF-α, IL-6 and MCP-1), and which may be related with the activation of NF-κB.
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Purpose To investigate the expression of forkhead box M1 (FOXM1) in human colorectal cancer(CRC) tissues,and to analyze the correlation between its expression and clinicopathological parameters and prognosis of CRC.Methods The expression of FOXM1 was examined in 297 cases of CRC tissues and 80 cases of normal peritumoral tissues by immunohistochemistry of SP.Western blot was used to detect the expression of FOXM1 in 20 cases of fresh CRC tissues and the corresponding normal peritumoral tissues.Results The expression of FOXM1 was significantly higher in CRC tissues (60.6％) compared with normal peritumoral tissues (17.50％,P ＜ 0.01).The positive expression of FOXM1 was related to the depth of invasion,lymph node metastasis,intravascular cancer embolus,distant metastasis and TNM stage (P ＜ 0.05).However,there was no correlation with age,sex,tumor location,tumor size,tumor differentiation degree,CEA and CA199 (P ＞0.05).The relative expression levels of FOXM1 protein was also significantly higher in CRC tissues (0.855 ± 0.063) compared with corresponding normal peritumoral tissues (0.150 ± 0.041,P ＜0.01).The 3-year overall survival of CRC patients with FOXM1 expression was significantly lower than that of patients without FOXM1 expression (P＜0.01),and FOXM1 is an independent prognostic factor for CRC.Conclusion The overexpression of FOXM1 protein in CRC tissues correlate with the clinicopathological characteristics and prognosis.FOXM1 may play an important role in the occurrence and metastasis of CRC,and it may be a new tumor marker and potential therapeutic target for CRC.
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AIM:To study the influence of lithium chloride (LiCl) on the neuronal differentiation of rat bone marrow mesenchymal stem cells (MSCs), and to explore whether autophagy was involved in this process .METHODS:MSCs were isolated and cultured in vitro.The cells were divided into LiCl group and control group .MSCs were treated withβ-mercaptoethanol as an inducer for triggering the cells to differentiate into neurons .The expression of neuronal markers-neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2), and autophagic marker-microtubule-associ-ated protein 1 light chain 3 ( LC3) were measured by immunofluorescence method and Western blot .An autophagy activator rapamycin and autophagy inhibitor 3-methyladenine (3-MA) were applied to modulate the autophagy in the LiCl treated-cells.The protein expression of NSE and MAP-2 were determined by Western blot .RESULTS: After induction, the ex-pression of NSE and MAP-2 were increased .The percentage of NSE-and MAP-2-positive cells and the expression of NSE and MAP-2 in the LiCl group were greater than those in control group (P<0.05).After induction, the number of LC3-positive dots and the expression of LC3-Ⅱin LiCl group were greater than those in control group (P<0.05).The expres-sion of NSE and MAP-2 increased when the autophagy was modulated by rapamycin in LiCl treated -cells, and on the contra-ry, the expression of NSE and MAP-2 were inhibited as autophagy was modulated by 3-MA.CONCLUSION: Lithium chloride may promote the neuronal differentiation of rat bone marrow mesenchymal stem cells by modulating autophagy .
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<p><b>OBJECTIVE</b>To understand the impact of effective interaction between public and media on the prevention and control of severe acute respiratory syndrome (SARS).</p><p><b>METHODS</b>Based on the theory of field epidemiology, the traditional and modern technologies were brought forward. In the late period of SARS epidemic, investigation on knowledge, attitude and practice on SARS prevention and control among general population was conducted.</p><p><b>RESULTS</b>During the 3 month period of SARS epidemic in Shenzhen, techniques of the "E-health and public health informatics" as an emerging fields were used to focus on population to eventually improve the health of entire population. Methods as television, radio, newspapers, magazines, printing and distribution of specific materials on SARS were included, and hotlines of telephone and mobile phone, web-based sites, electronic screening advertisements, and consultation or forum for the public etc. were used. Among 3405 subjects interviewed, more than 95% gave right answers on the SARS related questions. Most of them held the optimistic and scientific attitude. 80% of the population had taken the measures as recommended. 82.4% of the citizens were happy about the local government's action and 93.8% thought hospitals and healthcare workers should be respected and honored.</p><p><b>CONCLUSION</b>Dealing with the urgent public health problems and emerging infectious diseases outbreak, epidemiologists need to keep good relations with media, and making use of modern electronic technologies to communicate with the public.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , China , Epidemiology , Disease Outbreaks , Health Education , Health Knowledge, Attitudes, Practice , Infection Control , Methods , Information Dissemination , Methods , Mass Media , Public Health Informatics , Severe Acute Respiratory Syndrome , EpidemiologyABSTRACT
Five structural important residues of rice nonspecific lipid transfer protein LTP110 were mutated by site-directed mutagenesis. Sequence results showed that they were all mutated successfully. After trying various E. coli expression systems, thioredoxin fusion expression system was found to be a proper system to express wild type and mutant LTP110. cDNA sequences encoding wild type LTP110 and the mutants Y17A, P72L, R46A, D43A, C50A were cloned into two kinds of thioredoxin fusion expression vectors. The expression results were compared. In pTrxFus/GI724 expression system, wild type LTP110 and the mutants Y17A, P72L, R46A could be expressed at low level while D43A and C50A could not be expressed normally; in pET32a(+)/BL21 (DE3) trxB- expression system, wild type LTP110 and all mutant proteins could be expressed very well and the levels were higher than that in pTrxFus/GI724 system. LTP110 fusion protein expressed in pET32a(+) vector was purified and its activity was checked by fluorescence labeled fatty acid. Results indicated that the recombinant LTP110 fusion protein has lipid binding activity. This work provides good basis for the further study.