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Purpose@#Osteosarcoma (OS) universally exhibits heterogeneity and cisplatin (CDDP) resistance. Although the Wee1/CDC2 and nuclear factor кB (NF-κB) pathways were reported to show abnormal activation in some tumor cells with CDDP resistance, whether there is any concrete connection is currently unclear. We explored it in human OS cells. @*Materials and Methods@#Multiple OS cell lines were exposed to a Wee1 inhibitor (AZD1775) and CDDP to assess the half-maximal inhibitory concentration values. Western blot, coimmunoprecipitation, confocal immunofluorescence, cell cycle, and Cell Counting Kit-8assays were performed to explore the connection between the Wee1/CDC2 and NF-κB pathways and their subsequent physiological contribution to CDDP resistance. Finally, CDDP-resistant PDX-OS xenograft models were established to confirm that AZD1775 restores the antitumor effects of CDDP. @*Results@#A sensitivity hierarchy of OS cells to CDDP and AZD1775 exists. In the highly CDDP-tolerant cell lines, Wee1 and RelA were physically crosslinked, which resulted in increased abundance of phosphorylated CDC2 (Y15) and RelA (S536) and consequent modulation of cell cycle progression, survival, and proliferation. Wee1 inhibition restored the effects of CDDP on these processes in CDDP-resistant OS cells. In addition, animal experiments with CDDP-resistant PDX-OS cells showed that AZD1775 combined with CDDP not only restored CDDP efficacy but also amplified AZD1775 in inhibiting tumor growth and prolonged the median survival of the mice. @*Conclusion@#Simultaneous enrichment of molecules in the Wee1/CDC2 and NF-κB pathways and their consequent coactivation is a new molecular mechanism of CDDP resistance in OS cells. OS with this molecular signature may respond well to Wee1 inhibition as an alternative treatment strategy.
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OBJECTIVE：To study the improvement effects of Weijing deco ction on AECOPD model rats and its possibile mechanism. METHODS ：Totally 55 male SD rats were randomly divided into normal group ，model group ，Weijing decoction low-dose and high-dose groups （8.37，16.74 g/kg，by crude drug ），dexamethasone group （positive control group ，0.09 mg/kg），with 11 rats in each group. Except for normal group ，AECOPD model was induced by cigarettes combined with lipopolysaccharide in other groups. After modeling ，normal group and model group were given constant volume of water intragastrically ，and other groups were given relevant medicine intragastrocally ，twice a day ，for 14 days. After last intragastric administration ，the serum level of IL- 1 β was determined，and pathological changes of lung tissue and bronchus were observed in each group ；mRNA expression of MMP-9 and TIMP- 1 genes in lung tissue were detected ；protein expression of Ras homologous gene family member （RhoA）， dishevelled associated activator of morphogenesis- 1（DAAM1）and hyperplasic suppress gene （HSG）in lung tissue were also determined. RESULTS ：Compared with normal group ，the levels of IL- 1β in serum，mRNA expression of MMP- 9 and TIMP-1 as well as protein expression of RhoA and DAAM 1 in lung tissue were increased significantly in the model group（P＜0.05），while protein expression of HSG in lung tissue was decreased significantly （P＜0.05）；there were many chronic inflammatory cells infiltrating around the bronchus ，some airway mucosa epithelium exfoliating ，alveolar compensatory dilation，pulmonary septal capillary dilation and hyperemia. Compared with model group ，the levels of IL- 1β in serum，mRNA expression of MMP- 9 and TIMP- 1 in lung tissue were decreased significantly in Weijing decoction high-dose group （P＜0.05）；the protein expression of RhoA and DAAM 1 in lung tissue were decreased significantly in Weijing decoction low-dose and high-dose groups（P＜0.05），while the protein expression of HSG in lung tissue was increased （P＜0.05）；the pathological changes of Weijing decoction high-dose group ，such as inflammatory cells infiltrating around the bronchus and shedding of airway mucosa ， were improved significantly ， and there was complete alveolar epithelium structure but no obvious pulmonary dilation. CONCLUSIONS：Weijing decoction can improve AECOPD model rats to certain extent ；its mechanism may be associated with down-regulating mRNA expression of MMP- 9 and TIMP -1 as well as protein expression of RhoA and DAAM 1 in lung tissue ， up-regulating protein expression of HSG in lung tissue so as to inhibit the airway remodeling.
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Objective To construct a recombinant human adenovirus type 3 ( HAd3 ) vector ex-pressing one major epitope of dengue virus type 1.Methods The gene encoding the envelope protein (304-314 aa) of dengue virus type 1 was inserted into the hypervariable region 1 ( HVR1 ) of HAd3 hexon by using overlap PCR.The recombinant gene was cloned into the shuttle plasmid, then linearized with AsisⅠrestriction enzyme and co-transformed into Escherichia coli BJ5183 strains with the digested backbone plas-mid for homologous recombination.The recombinant plasmid pBRAdΔE3GFP-DENV1 was transfected into AD293 cells to rescue recombinant adenovirus strains (rAdΔE3GFP-DENV1).ELISA and Western blot as-say were performed to evaluate the humoral responses induced in BALB/c mice after the immunization with rAdΔE3GFP-DENV1 strains.Results The recombinant adenovirus strains were successfully rescued. ELISA and Western blot assay showed that the antibodies in serum sample could recognize dengue virus type 1 strains.Conclusion The recombinant adenovirus strains expressing the epitope of dengue virus type 1 were successfully constructed.This study provided evidence for the development of multivalent vaccines against dengue virus.
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OBJECTIVE:To investigate the effect of Changji’an capsules on abdominal pain and the molecular mechanism re-lated to calcitoningene-related peptide(CGRP)and corticosterone(CORT)in diarrhea-predominant irratable bowel syndrome(IBS-D)model rats. METHODS:After the rat models of IBS-D were established by the method of separation of breast mick combined with stimulation with acetic acid,the rats were randomly divided into model group(isometric normal saline),pinaverium bromide group(0.018 g/kg),and Changji’an capsules high,medium and low dose groups(2.812 g/kg,1.406 g/kg and 0.703 g/kg),and another SD rats were included in the normal control group(isometric normal saline). The drugs were given to the rats once a day for consecutive 14 d,ig. Injection of normal saline method was adopted to determine the rat’s sensibility to abdominal pain. The en-zyme-linked immunosorbent assay(ELISA)was adopted to determine the content of CORT in the serum of the rats. Subjected to re-verse transcription polymerse chain reaction(RT-PCR)was adopted to determine the expression of CGRP mRNA in the hypothala-mus and colon tissues of the rats. RESULTS:Compared with normal control group,threshold values of arching the back and stick-ing out the abdomen were decreased,the content of CORT in serum and expression of CGRP mRNA in the hypothalamus and co-lon tissues in model group were increased,with significant difference(P<0.01). Compared with model group,threshold values of arching the back and sticking out the abdomen were increased,the content of CORT in serum and expression of CGRP mRNA in the hypothalamus and colon tissues in pinaverium bromide group and Changji’an capsules high and medium dose groups were de-creased;the threshold value of arching the back in Changji’an capsules low dose group were increased,with significant difference (P<0.01 or P<0.05). CONCLUSIONS:Changji’an capsules can improve the abdominal pain in rats with IBS-D by a mechanism that may be related to the decrease in the expression of CGRP mRNA in the hypothalamus and colon tissues and the reduc-tion of the content of CORT in serum.
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Objective:To explore the reversal effect of Docetaxel and Capecitabine on rat breast precancerous from angiogenesis and expression of related regulatory factors VEGF mRNA in rats.Methods: 350 SD rats were divided into 7 groups.Model of rats mammary was induced by DMBA and was treated by Docetaxel and Capecitabine for 4 weeks.All of them were killed from 8th week.The microvascular were detected in the specimens, examination of histopathology was performed and the expression of VEGF mRNA was measured by in situ hybridization.Results:The rat mortality and the incidence of precancerous lesions increased obviously, and the incidence of precancerous lesion of the high-dose group and the middle dose reduced.The positive rate of the expression of MVD,TGF-αand VEGF mRNA tend to increase in all the groups.The positive-cell rate of VEGF mRNA of Docetaxel,Capecitabine, Docetaxel and Capecitabine in ADH was lower than in the model group,and the positive-cell rate of VEGF mRNA of Docetaxel and Capecitabine was lower than Capecitabine and Docetaxel separately.Conclusion: Combination of Docetaxel and Capecitabinecan decrease the expression of VEGF mRNA in precancerouslesion of rats mammary.
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Objective To establish a diarrhea rat model using multiple-stimulating factors and choosing the best indexes to assess whether the model is consistent with the disease characteristics of liver -QI stagnation with spleen deficien-cy in traditional Chinese medicine .Methods Newborn SD rats were randomly divided into model group ( n=20 ) and control group (n=10).The rats of model group were stimulated by maternal separation , restraint stress and rectum acetic acid irritation, while the rats in control group were fed as normal .Weight changes, rectal sensitivity, Bristol scores and wa-ter content of feces and histology of the colon tissues were used as evaluation indexes to assess whether the model meets the demands for further studies .Results The rats in the model group showed loss of appetite , increase of water intake and u-rine reduction .Some rats showed increased activity , and even mania .Bristol scores and water content of feces were signifi-cantly higher than that of the control group , and the rectal sensitivity was significantly increased .The colon mucosa showed slightly thickened submucosal layer and mild inflammatory cell infiltration in the model rats .Conclusions The rat model established in this study is better to simulate the clinical manifestation of liver -QI stagnation with spleen deficiency in Chi-nese medicine , and may meet the demands of related researches of this disease .
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Objective To evaluate narrow band imaging (NBI) assisted argon plasma coagulation (APC) in treatment of Barrett's esophagus (BE). Methods Suspected BE lesion was observed under white light, NBI and magnification, biopsies were taken at the site with characteristic pit pattern and capillary architecture of BE. A total of 86 patients with pathologically confirmed BE were randomly divided into NBI group (n= 42) to receive APC under NBI, or control group (n= 44) to receive APC under whit light. For APC procedure, the probe was inserted through biopsy channel to reach 1 cm beyond the endoscope tip, and was located 1-2 cm from the lesion to assure safe use. All patients were followed up with endoscopy and biopsy at 3 and 6 months after APC, respectively. Results There was no significant difference between 2 groups in effective rate of BE mucosal eradication at 3 and 6 months after APC procedure (P > 0.05). Conclusion NBI assisted APC is safe and effective in eradication of BE epithelium, in reducing procedure time and in relieving of functional gastrointestinal symptoms related with BE.