1.Clinical advancement of hypercoagulability in patients with ovarian cancer
Zheng FENG ; Hao WEN ; Xiaohua WU
China Oncology 2015;(3):231-234
The incidence of venous thromboembolism in patients with ovarian cancer is much higher than other gynecologic cancers. Approximate 20%of ovarian cancer patients have hypercoagulable status during different phases of their disease. Ovarian cancer itself can induce hypercoagulability, but meanwhile the over activated coagulation system may promote disease progression. Coagulation system disorder is one of the most important prognostic factors in ovarian cancer. Recently, hypercoagulability becomes a hot spot in the ovarian cancer research ifeld. This article reviews the mechanism of hypercoagulability, its clinical implication and correlated treatment in ovarian cancer patients.
2.Biological characteristics of human amniotic fluid-derived embryonic mesenchymal stem cells isolated and cultured using the two-step method
Jianxun FENG ; Xiaolin LA ; Yan MA ; Xiaojuan BI ; Hao WEN
Chinese Journal of Tissue Engineering Research 2009;13(45):8854-8860
BACKGROUND:In vitro isolation and purity technique of stem cells mostly depends on the identification of cell surface marker,such as monoclonal antibody adherent spreading method,flow cell sorting method and immunomagnetic beads sorting method,but the operation was complicated and the price was high.OBJECTIVE:To observe the biological characteristics of human amniotic fluid-derived embryonic mesenchymal stem cells,which were isolated and cultured using the two-step method.DESIGN,TIME AND SETTING:The opening study was conducted at the Stem Cell Research Room of Xinjiang Medical University from March 2008 to March 2009.MATERIALS:Totally 10 amniotic fluid specimens were obtained from pregnant women who underwent prenatal diagnosis following 16-22 weeks of gestation or voluntarily induced abortion.With ultrasonic guidance,amniocentesis was performed to collect 20-40 mL amniotic fluid.METHODS:Human amniotic fluid-derived embryonic mesenchymal stem cells were isolated and cultured using the two-step method.Amniotic fluid was first centrifuged and incubated till spindle-shape cells were seen,with the presence of flbroblast-tike cell colonies.Supematant was moved to a new 25 cm~2 culture flask for further culture till spindle-shape fibroblast-like mesenchymal stem cell colonies.When 70% confluence,cells were digested,and incubated in α-MEM,supplemented with basic fibroblast growth factor,served as the first passage.MAIN OUTCOME MEASURES:Morphological changes in human amniotic fluid-derived embryonic mesenchymal stem cells of primary culture and subculture were measured.Karyotype,cycle,growth curve and colony formation ability of human amniotic fluid-derived embryonic mesenchymal stem cells were measured.Surface antigen and cytokine were examined using flow cytometry,immunofluorescence and RT-PCR.RESULTS:Human amniotic fluid-derived embryonic mesenchymal stem cells were successfully isolated and subcultured.During metaphase,primarily cultured amniotic fluid cells presented scattered spindle cells and flbroblast-like mesenchymal stem cell colonies every 7 days.Passaged cells completely adhered in 12 hours.Following 1 or 2 days of latent period,cells proliferated rapidly.About 90% confluence was observed following 6 or 7 days of culture.Cell arranged regularly,showing whirlpool-shape,radiated shape.Cells were spindle-shape,with unclear boundary.Chromosome karyotype of human amniotic fluid-derived embryonic mesenchymal stem cells was normal diploid.Growth curve showed "S" shape,but the two-step method reached a peak at (6.1±0.5) days,which was significantly rapid compared with the one-step method (7.2±0.6) days (P=0.035).Flow cytometry analyses showed that P3 cells at S phase took up (14±2.3)% using the two-step method,which was more than the one-step method (9.0±1.4)% (P=0.031).Low-density human amniotic fluid-derived embryonic mesenchymal stem cells were incubated for 7 days prior to cells formed scattered cell colonies.However,colony forming efficiency using the two-step method (15.0±2.3)% were significantly more than the one-step method (10.0±1.8)% (P=0.021).Flow cytometry results showed that human amniotic fluid-derived embryonic mesenchymal stem cells expressed CD44,CD29 and CD105,but were negatively for CD45,CD34,HLA-DR.Immunofluorescence suggested that Oct-4-positive cells were observed in amniotic fluid.However,the proportion of Oct-4-positive cells using two-step method (1.2±0.3)% was significantly greater than the one-step method (0.9±0.2)% (P=0.041).RT-PCR suggested that human amniotic fluid-derived embryonic mesenchymal stem cells obtained using the two methods expressed Oct-4.CONCLUSION:Human multipotent mesenchymal stem cells are present in human amniotic fluid.The two-step culture protocol could be a kind of high performance and simple protocol which may not interfere with the normal prenatal diagnosis procedure.
3.Chemical constituents of Dalbergia odorifera.
Hao WANG ; Wen-Li MEI ; Zhi-Kai GUO ; Zhan-Feng XIA ; Hui-Min ZHONG ; Hao-Fu DAI
China Journal of Chinese Materia Medica 2014;39(9):1625-1629
Fourteen compounds were isolated from Dalbergia odoriferae and purified by repeated column chromatography on silica and sephadex LH-20 gel and structurally identified by spectral analysis. These compounds were identified as 4, 9-dimethoxy-3-hydroxypterocarpan (1), medicarpin (2), 2', 4', 5-trihydroxy-7-methoxyisoflavone (3), 2', 3', 7-trihydroxy-4'-methoxyisoflavan (4), formononetin (5), 3, 8-dihydroxy-9-methoxypterocarpan (6), koparin (7), 3-hydroxy-9-methoxypterocarp-6a-ene (8), 2'-hydroxyformononetin (9), stevenin (10), 2', 7-dihydroxy-4', 5'-dimethoxyisoflavone (11), lyoniresinol (12), 2, 4-dihydroxy-5-methoxy-benzophenone (13) and neokhriol A (14). Compounds 1, 3, 4, 6, 8, 12 and 14 were isolated from this plant for the first time. Antibacterial activity assay showed that compound 4 had inhibitory effect on Ralstonia solanacearum.
Anisoles
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chemistry
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isolation & purification
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pharmacology
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Anti-Bacterial Agents
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chemistry
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isolation & purification
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pharmacology
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Benzophenones
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chemistry
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isolation & purification
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pharmacology
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Chromatography
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methods
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Dalbergia
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chemistry
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Dextrans
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Gels
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Isoflavones
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chemistry
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isolation & purification
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pharmacology
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Microbial Sensitivity Tests
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Naphthalenes
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chemistry
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isolation & purification
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pharmacology
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Plant Extracts
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chemistry
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isolation & purification
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pharmacology
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Pterocarpans
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chemistry
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isolation & purification
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pharmacology
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Ralstonia solanacearum
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drug effects
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growth & development
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Silica Gel
4.Research on Relationship of Serum Urea Nitrogen Level and Upper Gastrointestinal Bleeding
Tingting HAO ; Yanli WEN ; Guangrong DAI ; Yichao FENG ; Li ZHANG ; Hua LI ; Xiaopeng MA
Journal of Modern Laboratory Medicine 2017;32(3):86-88,91
Objective To examine characteristics of patients with blood urea nitrogen (BUN) levels higher and lower than the normal limit.Methods During January 2012 to January 2015,116 patients with upper gastrointestinal diseases were selected to study,according to the patient's blood urea nitrogen level,all the patients were divided into high BUN group and low BUN group,and there were 76 patients in the high BUN group,and 40 patients in low BUN group,compared the biochemical indices,gastrointestinal bleeding forrest grading and disease severity of the two groups,and univariate logistic regression analysis.Results The serum white blood cell count,blood urea nitrogen,creatinine and glycated hemoglobin levels in patients of high BUN group [(9 593±5 012)× 102/μl,368.1±162.3 mg/L,11.2±3.7 mg/L and 6.38±1.08%] were significantly higher than that of low BUN patients [(6 804 ± 2 087) × 102/μl,121.0 ± 39.3 mg/L,8.1 ± 3.2 mg/L and 5.51 ± 0.42 %] (t =3.645~12.659,all P<0.05),and the hemoglobin levels (87.3±35.1 g/L) of the patients in high BUN group was significantly lower than that of the low BUN patients (108.0 ± 31.2 g/L) (t=3.252,P=0.032).Logistic regression analysis showed the presence of hemoglobin and glycosylated hemoglobin levelst of wo groups of patients was significantly different (P<0.05),and showed that showed the highest correlation with BUN.Gastrointestinal bleeding forrest hierarchical data of the two groups of patients showed no significant difference (P>0.05).The proportion of patients with gastric ulcers of high BUN patients was significantly higher than that of the low BUN patients (x2 =39.655,P=0.000).Conclusion Patients with high expression of serum urea nitrogen had more severe upper gastrointestinal bleeding,and it is worthy of attention in the process of clinical diagnostic.
5.Application of dynamic pupillometer in the evaluation of pupillary light reflex in C57BL/6 mouse
Rui, LIU ; Lei, ZHANG ; Hao, ZHANG ; Wen-zhi, HU ; Feng, XIA ; Jia, GENG ; Zuo-ming, ZHANG
Chinese Journal of Experimental Ophthalmology 2013;(1):75-80
Background Pupillary light reflex has been widely used in the diagnosis and evaluation of visual system and nervous system diseases.However,in animal experiments,functional evaluation of the visual system and nervous system needs more advanced technology and are affected by many factors.Objective This study was to explore the use of the dynamic pupillometer in evaluating pupillary light reflex and to discuss the influence of brightness of stimulate on relevant curve parameters in C57BL/6 mouse.Methods Ten healthy SPF male C57BL/6 mice were collected in this experiment.White light of five luminance levels (2,8,32,128,256 cd/m2) was used to stimulate the mice following a 2-hour dark adaptation.The stimulation was given at the 60-second intervals,for a duration of 100 ms at every stimulation.An infrared camera and video capture card were used to capture digital images during the measuring process in a scotopic environment,at a speed of 60 frames per second.Measuring outcome was saved in the*.AVI format.A software that was developed by our group was used to determine pupil diameter and output pupillary light reflex curve offline.Pupil initial diameter (R1),constriction amplitude (CA),constriction velocity (CV),latency (T1),time for maximum velocity (T2),time for maximum constriction (T3),time for maximun con-striction to 10.1% R1 re-dilation (RT)and re-dilation velocity (RV)were assessed,and the correlations between luminosity and measuring parameters were analyzed using the Spearman rank correlation.The use of animals followed the Regulations for thd Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results R1 values showed no statistically significant difference among the 5 different luminosity groups(F=1.117,P=0.361).A positive linear correlation was found between stimulating luminosity and CA(r=0.508,P< 0.01),but negative correlations were seen between stimulating luminosity and CV or RV (r=-0.625,-0.609,P<0.01).T1 and T2 values in the 5 different luminosity groups were not statistically significant (F =0.202,P =0.936 ; F =1.584,P =0.195).The different levels of stimulating luminosity showed positive linear correlations with T3 and RT values (r =0.791,0.609,P< 0.01).Conclusions The dynamic pupillometer can quantitatively measure the pupillary light reflex of C57BL/6 mice.The pupillary light reflex dynamic curve parameters of mouse were affected by stimulus luminosity levels.These outcomes offer a basis for the application of the dynamic pupillometer system for measuring pupillary light reflex in animal models.
6.Construction of lentiviral vector of peroxiredoxin 2 gene mediating RNAi and its effects on the proliferation of SW480 cell
Jihong FENG ; Zhongxue FU ; Kunming WEN ; Weidong LU ; Hao WANG ; Wangsheng CHEN ; Jinbao GUO ; Shouru ZHANG
Chongqing Medicine 2015;(1):14-17,20
Objective To construct a lentiviral expression vector of peroxiredoxin2(PRDX2) RNA interference (RNAi) and to investigate the effect of siRNA of PRDX2 genes on the proliferation of human colonrectal cancer SW480 cell .Methods RNAi tar‐get sequences were designed and synthesized towards the PRDX2 gene sequences .The lentiviral vector pGC‐EGFP‐shPRDX2 was constructed and identified .The vector was transformed into SW480 cells ,and the transfection efficiency was evaluated by fluores‐cence microscopy .The expression of PRDX2 was detected with Quantitative real‐time PCR (qRT‐PCR) and Western blot in the transfected cells .Cell growth and colony forming ability were detected with MTT and plate cloning technique .Results PRDX2 gene lentiviral vector was successfully established and was proved by gene sequencing .The expression of PRDX2 in mRNA and pro‐tein was significantly reduced(P<0 .05) .The PRDX2 mRNA and protein expression in SW480 transfected with lentiviral were sig‐nificantly reduced (P< 0 .05) ,and the ability of growth and proliferation were significantly reduced(P< 0 .05) .Conclusion PRDX2 gene lentiviral vector could be a stable and reliable tool .The proliferation and growth of SW480 cells transfected by pGC‐EGFP‐shPRDX2 could be effectively suppressed ,which could facilitate further investigation of the roles of PRDX2 gene in the de‐velopment and progression of colorectal cancer .
7.A control study of video -assisted thoracoscopic surgery (VATS)and conventional thoracotomy in treatment of thoracic esophagus cancer
Jian LI ; Xiaoming BAI ; Yun FENG ; Juntao HAO ; Lifeng MENG ; Wen ZHOU ; Jun MA
Chinese Journal of Primary Medicine and Pharmacy 2016;23(10):1463-1465,1466
Objective To investigate the effect of video -assisted thoracoscopic surgery (VATS)and conventional thoracotomy in treatment of thoracic esophagus cancer,to provide the reference for clinical.Methods 90 cases of thoracic esophageal cancer in our hospital from January 2012 to January 2015 were chosen as the research subjects.They were randomly divided into observation group(application of VATS treatment)and the control group (application of traditional open chest surgery).The quantity difference,chest drainage,hospitalization time,complica-tions,recovery conditions and other indicators of bleeding were compared in the two groups.Results The operation time of the observation group[(267.6 ±76.5)min]was shorter than that of the control group[(324.4 ±87.6)min]. The amount of intraoperative bleeding[(235.3 ±79.5)mL],drainage volume[(327.5 ±95.2)mL]of the observation group were less than the control group[(398.2 ±98.3)mL and (752.6 ±156.4)mL].Postoperative hospitalization time[(12.2 ±3.2)d]of the observation group was shorter than the control group[(15.8 ±4.4)d].The differences were statistically significant (t =3.276,8.644,15.575,4.439,all P <0.05).There were no significant differences between two groups in recurrence and metastasis rate,mortality rate and total survival rate (χ2 =0.123,0.212, 0.212,all P >0.05).The complication rate of the observation group was lower than that of the control group,the difference was statistically significant (χ2 =4.865,P <0.05).Conclusion In the treatment of thoracic esophageal carcinoma,VATS and conventional thoracotomy surgery has good effect,and VATS has small injury,less complication.
8.Application of detection of clonal immunoglobulin heavy chain gene rearrangement in paraffin-embedded tissues from B-cell non-Hodgkin lymphomas.
Xin-Xia LI ; Yun-Zhao CHEN ; Feng LI ; Wen-Hao HU ; Hong-An LI ; Jin-Fang JIANG
Chinese Journal of Pathology 2007;36(2):126-127
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Child
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Child, Preschool
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DNA, Neoplasm
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genetics
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Female
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Gene Rearrangement, B-Lymphocyte, Heavy Chain
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Humans
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Immunoglobulin Heavy Chains
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genetics
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Infant
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Lymphoma, B-Cell
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diagnosis
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genetics
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Lymphoma, Large B-Cell, Diffuse
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diagnosis
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genetics
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Male
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Middle Aged
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Paraffin Embedding
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Polymerase Chain Reaction
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Young Adult
9.Sequence analysis for full length genomes of human enterovirus 71 strains isolated in Linyi, Shandong Province
Hongling WEN ; Shubin HAO ; Feng GAO ; Li ZHAO ; Luying SI ; Xiaojing YUAN ; Dongxu WANG ; Zhiyu WANG
Chinese Journal of Microbiology and Immunology 2011;31(7):603-608
Objective To isolate enterovirus 71 from a death children,and analyze whether the neurovirulence was related to the variation of nucleotide and amino acid. Methods Enterovirus 71 was isolated from throat swabs which were colleted from Shandong Linyi People's Hospital. The full length genome was sequenced by amplification with RT-PCR and sequencing of 9 overlapped gene fragments covering full length of the genomes. The nucleotide and amino acid sequenced was aligned by BLAST, Bioedit and MEGA 4. Results A strain of enterovirus 71 was isolated and named as SDLY107. The full length was 7405 bp. The results of homology analysis of overall nucleotide sequence showed that strain Fuyang. Anhui. P. R. C/17.08/2 had highest homology (98.6%)with strain SDLY107, and the homology was 80.0% between strain SDLY107 with prototype strain BrCr/70,and 86. 5% between strain SDLY107 with nerve strain MS/87. Phylogenetic analysis showed that the phylogeny was close between SDLY107 with some isolated strains from Chinese Mainland, such as Beijing, Henan, Guangxi, Sbenzhen, Lanzhou, Fuyang, Chongqing and Zhejiang strains, which was clustered for C4 subtype. The results of amino acid sequence analysis showed that there were 2 mutations, E947D and K1873R, for strain SDLY107. Conclusion SDLY107 belonged to C4 subtype, amino acid mutations E947D and K1873R of which may be relevant to the pathogenicity of EV71.
10.Construction of Peroxiredoxin 2 gene lentiviral expression vector and effect on colorectal cancer SW480 cells
Jihong FENG ; Zhongxue FU ; Kunming WEN ; Shouru ZHANG ; Weidong LU ; Hao WANG ; Xingye WU
Chinese Journal of Immunology 2014;(8):1078-1082
To construct the lentiviral vector containing Peroxiredoxin 2(Prdx2) gene and the colorectal cancer cell line stably transduced with Prdx 2-containing vector , so as to provide a useful tool for studying the role of Prdx 2 in colorectal cancer.Methods: Prdx2 was amplified by PCR and inserted into lentiviral expression vector Ubi-MCS-EGFP-IRES-Puromycin (GV218) to generate Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector.The inserted Prdx2 gene was verified by double enzyme digestion and DNA sequencing.Subsequently ,lentiviruses were produced and transduced into SW 480 cells.EGFP expression was examined under fluorescence microscopy ,the expression of Prdx2 was detected with qRT-PCR and Western blot.Cell growth and colony forming ability were detected with MTT and plate cloning technique.Results: The lentiviral Prdx2 expression vector was successful construc-ted.Overexpression of Prdx2 was verified in SW480 cells with LV-Prdx2 vector.Prdx2 promoted SW480 cell growth and colony forming ability(P<0.05).Conclusion:Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector is successfully constructed,and the SW480/LV-Prdx2 cell line with stable transduction of Prdx2 containing vector is established.Overexpression Prdx2 can significantly promote the proliferation of colorectal cancer SW 480 cells.