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Objective:To investigate the mechanism of neurofibromin 1 (NF1) in gallbla-dder cancer.Methods:The experimental study was conducted. Human gallbladder cancer cell lines, including GBC-SD, NOZ, SGC996, EH-GB1, ZJU0428, human embryonic kidneys cell line 293T and human cervical cancer cell line HELA, were cultured. The recombinant plasmids (mRFP-YAP1 FL-FLAG and eGFP-MYC-NF1 2650?2750-HA) were constructed for co-immunoprecipitation experiment. The truncated Yes associated protein 1(YAP1) and NF1 recombinant proteins were purified in vitro. The interaction between NF1 and YAP1 in vitro or in vivo were verified by isothermal titration calori-metry (ITC) assay, GST pull-down experiment, co-immunoprecipitation, immunofluorescence, laser confocal microscopy, and the expression of NF1 protein in different gallbladder cancer cell lines was verified by Western blot experiments. Observation indicators: (1) interaction between NF1 and YAP1 in vitro; (2) interaction between NF1 and YAP1 in cells; (3) expression of NF1 protein in different human gallbladder cancer cell lines. The dissociation constants were exported from ITC 200 software and represented as Mean± SD. Count data were represented as absolute numbers. Results:(1) Interaction between NF1 and YAP1 in vitro. ① Results of ITC assay showed that there was interac-tion between PPQY and YAP1-WW1, between PPQY and YAP1 (Amino acid residues 162?275), and the dissociation constants between PPQY and YAP1-WW1, between PPQY and YAP1(Amino acid residues 162?275) were (0.42±0.06)mmol/L, (0.69±0.14)mmol/L, respectively. ② GST pull-down results indicated that the target protein His-Sumo-YAP1 WW1 was obviouly observed in protein lane of reaction system between GST-PPQY recombinant protein and His-Sumo-YAP1 WW1, relative to the reaction system between GST protein and His-Sumo-YAP1 WW1. The target protein His-Sumo-YAP1 WW2 was obviouly observed in protein lane of reaction system between GST-PPQY recombinant protein and His-Sumo-YAP1 WW2, relative to the reaction system between GST protein and His-Sumo-YAP1 WW2. (2) Interaction between NF1 and YAP1 in cells. ① Co-immunoprecipitation results indica-ted that NF1 protein was observed in cell lysis solution which was incubated by FLAG gel beads and cotransfected with mRFP-YAP1 FL-FLAG and eGFP-MYC-NF1 2650?2750-HA. ② Immuno-fluorescence and laser confocal microscopy results indicated that YAP1 and NF1 with obvious fluorescence were co-localized in the cytoplasm of human gallbladder cancer NOZ cells. However, YAP1 with obvious fluorescence was localized in the nucleus of human gallbladder SGC996 cells and NF1 showed weak fluorescence. (3) Expression of NF1 protein in different human gallbladder cancer cell lines. Western blot results showed that with the expression level of NF1 protein in HELA cell line as the standard, the relative expression levels of NF1 protein in EH-GB1, GBC-SD, NOZ, SGC996, ZJU0428 cell lines were 1.28, 0, 1.01, 0, 0, respectively. Conclusion:NF1 affects the gallbladder cancer by directly acting on YAP1 protein.
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OBJECTIVE：To systematically evaluate clinical efficacy of L-carnitine in the treatment of male infertility ，and to provide evidence-based reference for clinical treatment of male infertility. METHODS ：Retrieved from CNKI ，Wanfang database ， VIP，CBM，PubMed，Embase and the Cochrane library ，randomized controlled trials （RCTs）about L-carnitine and other chemical drugs in the treatment of male infertility were collected during the inception to Apr. 12th，2020. After data extraction of included literatures and quality evaluation with modified Jadad scale ，Meta-analysis was conducted by using RevMan 5.3 software. RESULTS：A total of 8 RCTs were included ，with 520 patients. The results of Meta-analysis showed that compared with other chemical drugs ，L-carnitine could significantly enhance the semen volume [MD＝0.55，95％CI（0.20，0.91），P＝0.002] and sperm mortality rate [MD＝1.60，95％CI（0.50，2.69），P＝0.004] of male infertility patients ，with statistical significance. There was no statistical significance in sperm count [MD＝4.00，95％ CI（－3.15，11.15），P＝0.27]，the percentage of forward motile sperm [MD＝12.58，95％CI（－3.87，29.03），P＝0.13]，and the percentage of inducing pregnancy rate [OR＝0.85，95％CI（0.47，1.52），P＝ 0.58] of male infertility patients. CONCLUSIONS ：L-carnitine can significantly improve the semen volume and sperm mortality of male infertility patients ，and has the same effects as other drugs on improving sperm count ，percentage of forward motile sperm and percentage of inducing pregnancy rate.
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AIM: To investigate the effect of curcumin on oxidized low-density lipoprotein (ox-LDL)-induced injury of human aortic endothelial cells (HAECs).METHODS: HAECs were pre-treated with curcumin at different concentrations and then treated with ox-LDL.The cell viability was assessed by MTT assay.The cell proliferation ability was analyzed by EdU assay.ELISA was used to determine the concentrations of interleukin-6 (IL-6), transforming growth factor β1 (TGFβ1), high mobility group box-1 protein (HMGB1) and secretory receptor for advanced glycation end products (sRAGE) in the HAEC culture medium.The binding activity of peroxisome proliferator-activated receptor γ (PPARγ) was evaluated by electrophoretic mobility shift assay.The protein levels of HO-1, HMGB1, RAGE,IL-6,TGFβ1 and phosphorylated PPARγ in the HAECs were determined by Western blot.RESULTS: The viability and the proliferation ability decreased significantly in the HAECs treated with ox-LDL.The PPARγ/HO-1 signaling pathway was inhibited while its down-stream HMGB1/RAGE signaling pathway was activated by ox-LDL.The levels of IL-6, TGFβ1, HMGB1 and sRAGE were increased.Pre-treatment with curcumin activated PPARγ/HO-1 signaling pathway and inhibited HMGB1/RAGE signaling pathway in ox-LDL treated HAECs in a concentration-dependent manner.The levels of IL-6, TGFβ1, HMGB1 and sRAGE were also decreased dramatically by pre-treatment of curcumin in a concentration-dependent manner.CONCLUSION: ox-LDL induces HAEC damage by inhibiting PPARγ/HO-1 to activate HMGB1/RAGE inflammatory signaling.Curcumin exerts protective effect on ox-LDL treated HAECs via activating PPARγ/HO-1 signaling pathway.
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OBJECTIVE:To explore the effects of dual-phase insulin aspart with different dosing regimens on the related indica-tors of type 2 diabetic (T2DM) patients with poor glycemic control. METHODS:70 T2DM patients with poor glycemic control were randomly divided into group A and group B. All patients were given metformin and stopped other antidiabetic drugs;based on it,group A was additionally given Dual-phase insulin aspart injection,0.5 U/(kg·d),in the morning and evening before a meal by subcutaneous injection;group B was given Dual-phase insulin aspart injection,0.5 U/(kg·d),once before lunch time for 6-10 U and other twice in the morning and evening before a meal by subcutaneous injection. Both groups were treated for 12 weeks. Glu-cose control rate,glucose control time,and glucose indicators,daily fluctuations of glucose before and after treatment and incidenc-es of hypoglycemia and adverse reactions in 2 groups were observed. RESULTS:Glucose control rate in group B was significantly higher than group A,glucose control time was significantly shorter than group A,incidence of hypoglycemia was significantly low-er than group A(P<0.05). After treatment,glucose indicators and daily fluctuations of glucose in 2 groups were significantly lower than before,and group B was lower than group A(P<0.05). There were no obvious adverse reactions during treatment. CONCLU-SIONS:Conpared with 2 times a day,Dual-phase insulin aspart with 3 times a day for administration can effectively improve the glucose control rate and glucose levels in the treatment of T2DM patients with poor glycemic control,with good safety.
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Objective:To evaluate the efficacy and safety of capecitabine-based regimens in treating patients with liver metastases from breast cancer. Methods:A total of 163 patients with liver metastases from breast cancer who received capecitabine-based regimens between January 1, 2000 and Dec 31, 2011 were retrospectively reviewed. The clinicopathological characteristics and treatment outcomes of these patients were evaluated. Results:Of the 163 patients retrospectively analyzed, 109 received docetaxel plus capecitabine (TX) and 54 received vinorelbine plus capecitabine (NX). TX treatment achieved objective responses in 59 patients (54.1%), including complete response in four patients, partial response in 55 patients, and stable disease in 25 patients. In patients who received NX, the objective response and clinical benefit rates were 50.0%and 70.4%, respectively;one patient had a complete response, 26 patients had a partial response, and 11 patients had a stable disease. The safety profiles of TX treatment were more favorable and predictable compared with NX treatment, with a low incidence of grade 3/4 adverse events in hematological and non-hematological toxicities. Results showed that median overall survival after liver metastases (LMS), progression-free survival (PFS), and post-metastasis survival (MSR) were 26, 8, and 28 months in the TX arm. LMS, PFS, and MSR were longer in the TX arm than in the NX arm. Conclusion:Capecitabine-based regimens showed tolerance in patients with liver metastases from breast cancer. TX treatment had a tendency of lower toxicity and was more effective compared with NX treatment.
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Objective To study the effect of camphor oil in killing Demodex in vitro and to further analyze the killing mechanism.Methods The mites were collected with adhesive cellophane tape and randomly divided into six groups.The killing effect of camphor oil was investigated with different concentrations against Demodex in vitro.The reaction progress was pictured using Motic Images software.Results Camphor oil had better killing effect on Demodex folliculorum than on D.brevis in vitro.The death rate increased with the drug concentration and time.The most suitable and effective concentration of killing both Demodex folliculorum and D.brevis in vitro was 12.5%.After the drug was given,the mites contracted and twisted,and then relaxed and died.Conclusion Camphor oil has strong killing effect on Demodex in vitro.The main mechanism of camphor oil may be related to direct contact and neuromuscular toxicity.