Objective To study the mechanism of overexpression of retinoic acid receptor alpha( RARα)in attenuating renal interstitial fibrosis(RIP)in rats. Methods Porty 6_week_old male SD rats were randomly divided into 4 groups:sham operation group,model group,negative control group and transfection group,with 10 rats in each group. Rats in model group were separated and double ligated with left ureter;rats in sham operation group were not li_gated with ureter;rats in transfection group and negative control group were transfected with adeno_associated virus and negative control virus carrying RARα gene on the basis of model group,respectively. All rats were sacrificed 2 weeks later. Left kidney tissues were taken for pathological examination and RIP index was calculated. The expression of colla_genⅣ(Col_Ⅳ)and fibronectin(PN)in renal tissue was detected by using immunohistochemistry. The expressions of RARα,prohibitin(DHB)and transforming growth factor_beta 1(TGP_β1)in renal tissue were detected by using real_time fluorescence quantitative polymerase chain reaction( RT _qDCR)and Western blot. Results (1)Com_pared with sham operation group,the RIP index was significantly increased in model group(22. 81 ± 2. 43 vs. 2. 34 ± 0. 55,q﹦24. 94,P〈0. 05);compared with model group,the RIP index was not of significant difference in negative control group(22. 81 ± 0. 43 vs. 22. 26 ± 3. 43,q﹦0. 67,P〉0. 05),however it significantly decreased in transfection group(14. 06 ± 2. 99 vs. 22. 81 ± 2. 43,q﹦10. 66,P〈0. 05).(2)Compared with sham operation group,the mRNA and protein expressions of RARα,DHB significantly decreased in model group,but TGP_β1 mRNA and protein,Col_Ⅳand PN protein expression significantly increased in model group( mRNA:0. 43 ± 0. 17 vs. 1. 00 ± 0. 00,0. 34 ± 0. 08 vs. 1. 00 ± 0. 00,2. 97 ± 0. 54 vs. 1. 00 ± 0. 00,all P〈0. 05;protein:0. 25 ± 0. 10 vs. 0. 51 ± 0. 06,0. 24 ± 0. 07 vs. 0. 58 ± 0. 04,0. 59 ± 0. 09 vs. 0. 33 ± 0. 06,16. 01 ± 0. 87 vs. 8. 79 ± 0. 39,14. 64 ± 0. 32 vs. 9. 36 ± 0. 59,all P〈0. 05);com_pared with model group,the mRNA and protein expressions of RARα,DHB,TGP_β1 and Col_Ⅳand PN protein ex_pression had no significant difference in negative control group(all P〉0. 05);compared with model group,the mRNA and protein expression of RARα,DHB mRNA and protein expression significantly increased,but the TGP_β1 mRNA and protein,Col_Ⅳ and PN protein expression significantly decreased in transfected group( mRNA:0. 86 ± 0. 07 vs. 0. 43 ± 0. 17,0. 89 ± 0. 11 vs. 0. 34 ± 0. 08,1. 65 ± 0. 28 vs. 2. 97 ± 0. 54,all P〈0. 05;protein:0. 40 ± 0. 07 vs. 0. 25 ± 0. 10,0. 45 ± 0. 10 vs. 0. 24 ± 0. 07,0. 43 ± 0. 08 vs. 0. 59 ± 0. 09,11. 57 ± 0. 33 vs. 16. 01 ± 0. 87,11. 67 ± 0. 53 vs. 14. 64 ± 0. 32,all P〈0. 05).(3)Correlation analysis revealed that RARα protein expression was negatively correlated with RIP index,Col_Ⅳ,PN,TGP_β1(r﹦ _0. 78,_0. 78,_0. 76,_0. 76,all P〈0. 05);DHB protein expression was negatively correlated with RIP index,Col_Ⅳ,PN,TGP _β1( r ﹦ _0. 87,_0. 87,_0. 88,_0. 75,all P 〈0. 05);RARα protein was positively correlated with DHB(r﹦0. 85,P〈0. 05). Conclusion Overexpression of RARα could attenuate RIP by enhancing DHB expression in rats subjected to unilateral ureteral obstruction.

Objective: To investigate the regulatory effect of miR-1297 on the malignant biological behaviors of breast cancer cells and its underlying mechanism. Methods: Twenty pairs of breast cancer tissues and para-cancer tissues resected at the Department of Thyroid and Breast Surgery of Leshan People′ s Hospital from May 2016 to May 2018, as well as breast cancer cell lines MCF-7, SW626, HCC1937 and human breast epithelial MCF-10A cells were collected for this study. qPCR was performed to evaluate the expression of miR-1297 in breast cancer tissues and cell lines. The experimental cells were divided into control group, miR-1297 inhibitor group; TET3 over-expression group and simultaneous over-expression of TET3 and miR-1297 group. CCK-8 assay was used to detect the cell proliferation of MCF-7 cells; Transwell assay was carried out to detect the migration and invasion of MCF-7 cells; and WB was used to measure the expressions of TET3 and EMT related proteins (E-cadherin, N-cadherin and vimentin). Dual luciferase reporter gene assay was used to verify the relationship between miR-1297 and TET3. Results: miR-1297 was up-regulated in both breast cancer tissues and cell lines (P<0.01 or P<0.05). Knockdown of miR-1297 dramatically repressed the proliferation, migration, invasion and EMT of MCF-7 cells (P<0.01 or P<0.05). Over-expression of TET3 significantly up-regulated the expression of TET3 in MCF-7 cells (P<0.05). Simultaneous over-expression of TET3 and miR-1297 could reverse the expression level of TET3 in MCF-7 cells and the inhibitory effect of TET3 on the proliferation, migration, invasion and EMT of MCF-7 cells. Dual luciferase reporter gene assay results showed that miR-1297 targetedly bound to the 3' UTR of TET3. Further experiment results demonstrated that miR-1297 targetedly down-regulated TET3 and promoted the malignant biological behaviors of MCF-7 cells. Conclusion: miR-1297 is up-regulated in breast cancer tissues and cells; it promotes the malignant biological behaviors such as proliferation, migration, invasion and EMT through targetedly down-regulating the expression of TET3.

Objective To explore the role of c-Jun N-terminal protein kinase(JNK)pathway in renal tubular epithelial cells(RTEC)transdifferentiation by detecting the expression of JNK pathway in the injury of hypoxic RTEC. Methods In vitro cultured rat RTEC were randomly divided into four groups of normal control, hypoxia,inhibitors,dimethyl sulfoxide(DMSO). The groups of hypoxia,inhibitor and DMSO were placed into a vacuum tank to establish hypoxia model. The cells were harvested 6,12 and 24 h after hypoxia started. RT-PCR and Western blot test were used to detect the mRNA expressions of alpha smooth muscle actin(α- SMA)and protein expressions of JNK,pJNK and alpha SMA,respectively,in all the groups. Results Hypoxia induced significantly increased expressions of α-SMA mRNA and proteins,and JNK and pJNK proteins as well in RTEC (all P<0.05).After addition of inhibitors,the expression of α-SMA mRNA and proteins,the protein expression of pJNK in RTEC were significantly decreased(all P < 0.05)and the expression of JNK protein was significantly increased(P < 0.05). There was no significant difference in the expressions of JNK,pJNK,α-SMA in RTECs between hypoxia group and DMSO group(all P>0.05).There was a significant positive correlation between pJNK protein and α-SMA protein expression(P < 0.01). Conclusion In hypoxic RETC injury in rats,JNK pathway may be involved in the RTEC phenotype transdifferentiation.

Objective To explore the effect of overexpression of retinoic acid receptor α(RARα)on epithelial-to-mesenchymal transition(EMT)induced by hypoxia in renal tubular epithelial cells(NRK-52E).Methods The RARα lentivirus vector and negative control lentivirus vector were synthetised.The NRK-52E cells were divided into 4 groups:the normal control group,the hypoxia model group,the transfection group and the negative control group.Puro-mycin(2 mg/L)was added in transfection group and negative control group for screening after gene interference for 72 h.Then the 2 groups were subjected to hypoxia/reoxygenation,but the normal control group had no treatment. The change of cellular morphology was observed by using light microscope;the mRNA and protein expressions of RARα, E-cadherin,α -smooth muscle actin(α-SMA)in NRK-52E cells were detected by adopting reverse transcription-polymerase chain reaction(RT-PCR)and Western blot after hypoxia for 48 h.Results (1)Light microscope re-vealed that cells in both hypoxia model group and negative control group cells became atrophic and elongated,which were consistent with the morphology of myofibroblasts.But cells in transfection group cells were cubic,forming an epi-thelial monolayer.(2)Compared with the normal control group,the mRNA and protein expressions of RARα and E-cadherin in hypoxia model group were dramatically reduced(mRNA:0.58 ± 0.12 vs.1.00 ± 0.00,0.11 ± 0.00 vs. 1.00 ± 0.00,t= -0.63,767.30,all P<0.05;protein:0.63 ± 0.12 vs.1.62 ± 0.16,0.44 ± 0.22 vs.1.27 ± 0.08,t=8.61,6.19,all P<0.05),but the mRNA and protein expressions of α-SMA were higher(3.47 ± 0.83 vs.1.00 ± 0.00,1.39 ± 0.16 vs.0.64 ± 0.10,t= -5.01,-6.91,all P<0.05).(3)The mRNA and protein expressions of RARα and E-cadherin in the transfection group were significantly increased,compared with hypoxia model group(mRNA:4.69 ± 1.34 vs.0.58 ± 0.12,0.23 ± 0.00 vs.0.11 ± 0.00,q=9.13,25.48,all P<0.05;protein:1.39 ± 0.19 vs. 0.63 ± 0.12,0.87 ± 0.09 vs.0.44 ± 0.22,q=7.92,4.30,all P<0.05)and negative control group(mRNA:4.69 ± 1.34 vs.0.55 ± 0.21,0.23 ± 0.00 vs.0.12 ± 0.01,q=9.20,23.35,all P<0.05;protein:1.39 ± 0.19 vs.0.65 ±0.18,0.87 ± 0.09 vs.0.39 ± 0.21,q=7.71,4.80,all P<0.05).Conversely,the mRNA and protein levels of α-SMA were obviously lower in transfection group(1.52 ± 0.34 vs.3.47 ± 0.83,4.05 ± 0.81,0.82 ± 0.13 vs.1.39 ± 0.10,1.17 ± 0.10,q=4.88,6.33,7.50,4.61,all P<0.05).The difference in mRNA and protein expressions of RARα,E-cadherin,α-SMA between the hypoxia group and the negative control group had no statistical significance (all P>0.05).Conclusion Overexpression of RARα could alleviate EMT of renal tubular epithelial cells induced by hypoxia.

Objective To observe the effect of calycosin on cisplatin?induced inhibition of human gastric BGC823 cells. Methods BGC823 cells were treated with different drugs:saline,cisplatin,calycosin and cisplatin combined with calycosin. MTT assays were used to detect the prolif?eration rate of BGC823 cell. Then the protein level and RNA level of cyclin D1,CDK4 and CDK6 were detected by Western blotting and Real?time PCR. Results The proliferation inhibition rates of BGC823 cells treated with 20μg/mL cisplatin,10μg/mL meperoflavone,and combina?tion of the 2 drugs were 56.44%± 2.08%,9.52%± 2.77%and 74.44%± 0.82%,respectively. The inhibition rate of the combination of drugs was significantly higher than that of the single drug treatment group(P<0.05). In addition,we found that calycosin can significantly enhance the inhi?bition of Cyclin D1,CDK4 and CDK6 by cisplatin in protein level and RNA level. Conclusion Calycosin can significantly increase the inhibitory rate of cisplatin on BGC823 cell proliferation,and the combination of the two drugs can reduce the side effects of cisplatin.

Objective To determine the efficiency of microwave heating of the conjunctival sac flushing liquid combined with disposible blood transfusion device to prepare for laser-assisted in situ keratomileusis (LASIK).Methods A total of 150 healthy myopia patients undergoing LASIK in Myopia Treatment Center of our hospital from February to August 2013 were divided into two groups randomly according to operation date,75 patients in each group.The observation group used a microwave oven to heat the flushing liquid connected with a disposable blood transfusion device.The control group used a hot water bath to heat the flushing liquid,which was then connected to a disposable infusion device.The heated liquid by both methods was applied to conjunctival sac flushing before LASIK.The time required for liquid heating was recorded and the outcome of the conjunctival sac flushing was compared between the two groups.Results The time required by the observation group and the control group of heating flushing fluid from 5,10,15,20,25 and 30 ℃ to (36.0±1.0) ℃ was 124,104,84,64,44,24 s and 372,312,252,192,132,72 s,respectively.The conjunctival sac flushing effect comparison between the observation group and the control group was as follows:in four indicators such as the operation cooperation,conjunctiva congestion,spasm of eyelid,eye pain,there was no significant difference between two groups; for tear fat deposition,there was significant difference between two groups (4 cases vs.15 cases,x2=7.292,P ＜ 0.01).Conclusions Microwave heating of the conjunctival sac flushing liquid connected with a disposable blood transfusion device is efficient,safe and easy to operate.Therefore,it gains approvement and high opinion of medical workers and is worth being popularized in clinic.

Objective To observe the therapeutic effect of periodical treatment of Sanjiexiaopi decoction on hyperplasia of mammary gland during premenstrual phase. Methods 110 cases were randomly recruited into a treatment group and a control group with 55 case in each group. The treatment group was given Sanjiexiaopi decoction 7 days before menstruation, and the control group was given Rupixiao tablet. Both groups received no treatment during menstruation and were treated for three months. Patients in both groups underwent molybdenum target X-ray photographs and infrascan before and after the treatment. Therapeutic effects of the two groups.were compared. Results The total effective rate was 96.4% and 81.8% in the treatment group and the control group respectively. The patients in treatment group showed obvious improvent than the control group in terms of X-ray photographs and infrascan, having statistical significance (P<0.05) . Conclusion Sanjiexiaopi decoction has obvious effects on hyperplasia of mammary gland during premenstrual phase.

AIM and METHODS:The Ph.D.-7 phage display library was used to isolate peptides specific for glioma SWO-38 cell by whole cell screening.Moreover,binding efficiency analysis was carried out to test the binding specificity of the clones obtained. RESULTS: After three rounds of biopanning,a high concentration of phage clones was obtained and two of them were found to be highly specific to glioma SWO-38. CONCLUSION: Highly specific clones against neurtral glioma cells can be obtained from a phage display library by simple procedures.

It has long been acknowledged that there is a link between inflammation and cancer, but its molecular mechanism remains unclear. A key player in inflammation is nuclear transcription factor NF-?B, that activity is triggered in response to infectious agents and proinflammatory cytokines via the I?B kinase. In parenchyma cells, inflammation through I?B kinase/NF-?B pathway suppresses apoptosis, accelerates cell cycle, then promote tumorigenesis. In mesenchyma cells inflammation through I?B kinase/NF-?B pathway produces cytokines and chemokines that may serve as tumor growth factors. To sum up, I?B kinase/NF -?B pathway represents a critical molecular link between inflammation and cancer.

AIM: Three different antisense oligonucleotides complementary to basic fibroblast growth factor (bFGF) mRNA were compared in inhibitory effect on gene targeted expression. METHODS: After transfecting bFGF antisense oligonucleotides (asODN) into SWO-38 cells by lipofectin,the proliferation of cells was identified by MTT method, apoptosis was examined by flow cytometric cell cycle analysis and the expression levers of bFGF were detected by Western-blotting. RESULTS: There were 49%,33%,51% inhibition of cell growth and 35%, 27%, 18% cell apoptosis after asODN1, asODN2 and asODN3 treatment.In addition,the decrease in bFGF protein was 63%, 42%, 11%, respectively. CONCLUSION: The data suggeste that asODN1 is a potent target to bFGF mRNA, which inhibits cell growth and induces apoptosis in SWO-38 cells.