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Objective:To observe the clinical effect of hormone replacement therapy(HRT) on patients with cervical squamous cell carcinoma after operation and its effect on sex hormone, blood fat, bone content and tumor markers, and to evaluate the clinical significance of HRT therapy.Methods:From January 2016 to January 2019, a total of 100 patients with cervical squamous cell carcinoma admitted to Zhuhai Maternal and Child Health Hospital were analyzed retrospectively.After surgical treatment, 50 patients were willing to receive hormone replacement therapy as the experimental group and 50 patients received routine treatment as the control group, Kupperma score was used to evaluate the severity of perimenopausal symptoms, and the serum levels of sex hormones, blood fat, bone content and tumor markers in the two groups before and after treatment were detected and the occurrence of adverse reactions were evaluated.Results:Before treatment, there was no significant difference in kupperma score, estradiol, follicle-stimulating hormone, luteinizing hormone level, lipid level, bone content and tumor marker level between the two groups(all P>0.05). After HRT treatment, kupperma score in the experimental group was significantly lower than that before treatment((11.47±5.12) vs.(20.46±7.52), t=7.262). In the detection of sex hormones, the levels of estradiol in the experimental group was significantly increased((39.26±7.43) ng/L vs.(13.78±7.52) ng/L, t=12.345), and the levels of follicle stimulating hormone((34.25±7.61) U/L vs.(62.18±19.12) U/L, t=8.245)and luteinizing hormone((20.31±6.25) U/L vs.(35.08±6.27) U/L, t=5.452, P=0.004) in the experimental group were significantly decreased.In the detection of blood lipid level, bone content and tumor markers, high-density lipoprotein((1.62±0.33) mmol/L vs(1.34±0.26) mmol/L, t=4.592, P=0.008) and alkaline phosphatase levels((66.21±25.75) U/L vs.(46.88±9.06) U/L, t=5.912, P=0.001)was significantly increased, the low density lipoprotein((2.78±0.43) mmol/L vs.(2.87±0.78) mmol/L, t=4.265, P=0.012)and total cholestenone((4.02±0.45) mmol/L vs.(4.23±0.91) mmol/L, t=5.761, P=0.002) levels were significantly decreased in the experimental group, with statistically significant differences(all P<0.05). There was no significant difference in the level of serum SCC antigen between the experimental group and the control group before and after treatment ( P> 0.05); there was no significant difference in the above indicators between the experimental group and the control group before and after treatment ( P> 0.05); there was no significant increase in adverse reactions such as breast swelling and pain (3 cases), body mass increase (2 cases), bleeding (1 case), body pain (4 cases), vomiting (4 cases), etc. between the experimental group and the control group (2, 1, 2, 3, 1 case) There was no statistical significance ( P> 0.05). Conclusion:HRT treatment of cervical squamous cell carcinoma patients after surgery can significantly improve the peri menstrual syndrome caused by low estrogen level, and did not significantly increase the risk of recurrence of cervical cancer patients and adverse reactions.
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Objective To explore the best technology to produce recombinant mouse endostatin by thioredoxin fusion expression system.Methods The recombinant plasmid pThioHis-endo was further transformed into different E.coli.,including BL21,JM109,DH5?.After induction with IPTG of different concentration or for different time period,thioredoxin-endo fusion protein was expressed in E.coli.and the product was identified by SDS-PAGE.The recombinant endostatin expression and extraction,wash,degeneration and refolding of inclusion bodies were carried out by the optimized route,and the product was further purified by affinity chromatography through Ni~(+) column and identified by SDS-PAGE.Results No difference of endostatin expression in BL21,JM109,DH5? was found.The optimal concentration of IPTG was 0.9 mmol/L and optimal inductive phase was 5 h.The soluble recombinant endostatin could be obtained by affinity chromatography through Ni~(+) column.Conclusion Soluble endostatin recombinant fusion protein of high purity and yield could be obtained by the optimized technique route.
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AIM: To investigate the changes of cytosolic free calcium concentration ([Ca 2+ ] i) and expression of Bcl-2 in HL-60 cells treated by 6F isolated from Pteris semipinnata L. (PSL), and to discuss the relations between calcium ion and cytotoxicity and DNA fragment induction effects of 6F. METHODS: HL-60 cells were used as in vitro model. [Ca 2+ ] i was measured on fluorescent spectrophotometry using Fura-2/AM as Ca 2+ indicator. Bcl-2 expressing level was measured by flow cytometry. Tetrazolium salt (MTT) and diphenylamine staining methods were applied for cytotoxicity assay and DNA fragmentation detection, respectively. RESULTS: [Ca 2+ ] i increased obviously in a dose and time dependent manner after treated HL-60 cells with 6F. 6F decreased the expressing level of Bcl-2. Adding 2 mmol/L Ca 2+ to the medium, or 1 mmol/L EDTA to chelate Ca 2+ , or 4 ?mol/L calcium ionophore A 23187 to increase the concentration of cytosolic Ca 2+ , the DNA fragment induction by 6F was not affected, whereas the cytotoxicity of 6F was enhanced. 250 ?mol/L Zn 2+ attenuated the DNA fragment induction, and the cytotoxicity of 6F against HL-60 cells was enhanced significantly. CONCLUSION: It was speculated that the decreased expressing of Bcl-2 by compound 6F was related to increased [Ca 2+ ] i in HL-60 cells, and DNA fragment induction was possibly catalyzed by Ca 2+ - independent DNase.
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AIM To investigate the effects of tranfection of IL 2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS Spleen cells were transfected with IL 2R antisense RNA eukaryotic expression plasmids using adhesion assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL 2R mRNA and protein expression level were measured by slot blot hybridization assay and flow cytometry method respectively. RESULTS The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti mIL 2R?? and pciAnti mIL 2R?? transfected group was higher than that of pcAnti mIL 2R? and pcAnti mIL 2R? transfected group; the inhibitory rate of pcAnti mIL 2R? tranfected group was higher than that of pcAnti mIL 2R? tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL 2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION IL 2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL 2R?? chimeric antisense RNA showed higher inhibitory rate than ? or ? antisense RNA. IL 2R? antisense RNA was more effective than ? antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL 2R antisense RNA was exclusively on the growth of cells functionally expressing IL 2R. The inhibitory effect on the spleen cells proliferation was likely due to the blocking of IL 2R expression by antisense RNA.