1.Exploration on the mechanism of Xuanfei Dahe Decoction in the treatment of asthmatic mice based on network pharmacology and experimental validation
Mengting DAI ; Youlan LIN ; Huan YU ; Mengqing WANG ; Yunfei SHUAI
International Journal of Traditional Chinese Medicine 2025;47(9):1264-1271
Objective:To explore the mechanism of Xuanfei Dahe Decoction in treating asthmatic mice through network pharmacology and animal experiments.Methods:The active components and their targets of Xuanfei Dahe Decoction were retrieved from TCMSP, and the asthma targets were obtained by searching the GeneCards, DisGeNet, OMIM, TTD and DrugBank databases. PPI network of intersection targets was constructed using string database and Cytoscape 3.9.0 software. Go function and KEGG pathway enrichment were analyzed by metascape database. The mice were divided into a blank group of 6 mice and a model group of 30 mice according to the random number table method. The asthma model was prepared in the model group. Totally 30 successfully modeled mice were divided into the model group, the dexamethasone group, and Xuanfei Dahe Decoction low-, medium- and high-dosage groups according to the random number table method, with 6 mice in each group. On the 5th day, gavage was initiated. Xuanfei Dahe Decoction low-, medium- and high-dosage groups were respectively gavaged with Xuanfei Dahe Decoction liquid at concentrations of 6.84, 13.68 and 27.36 g/kg. The dexamethasone group was gavaged with dexamethasone acetate tablets at concentrations of 2.73 mg/kg. The blank group and the model group were gavaged with the same volume of sterile normal saline once a day for 14 consecutive days. The pathological changes of lung tissue were observed by HE staining, the level of serum IL-17 was detected by ELISA, and the expression of IL-17 in lung tissue was detected by immunohistochemistry.Results:120 asthma targets and 13 key targets were obtained from Xuanfei Dahe Decoction. Pathway enrichment analysis suggested that IL-17 signaling pathway was one of the key pathways. Compared with the model group, the levels of IL-17 in the serum of the low-dose, medium-dose and high-dose Xuanfei Dahe Decoction groups and the expression of IL-17A in the lung tissue of the medium-dose Xuanfei Dahe Decoction group decreased ( P<0.05). Conclusion:Xuanfei Dahe Decoction may treat asthma by restrain airway inflammation mediated by Th17/IL-17.
2.Analysis on allergic diseases from the "lung being responsible for regulating visceral activities"
Huan YU ; Wenyuan GUO ; Yunfei SHUAI ; Leqin XIONG ; Mengqing WANG
International Journal of Traditional Chinese Medicine 2025;47(12):1664-1669
The symptoms of allergic diseases are complex and changeable, and the pathogenesis is complex and difficult to distinguish, and the prevalence of allergic diseases has gradually increased in recent years. Due to the relative limitations of Western medical treatment, in order to further meet the clinical treatment goals, the "lung being responsible for regulating visceral activities" is now used as the theoretical basis, combined with modern medicine, to explore the pathogenesis and etiological mechanism of this disease. Based on the classics, it is proposed that allergic diseases have four characteristics: "onset of allergies, seasonality of disease onset, diversity of symptoms, and recurrence of recovery", and summarized the etiology and pathogenesis as "stagnation of incubative pathogenic factors and disharmony between the nutritive and defensive levels, imbalance of waterways and obstruction of phlegm and dampness, a dysfunction in the pivotal qi flow leading to malnourishment of the skin and hair, the intermingling of cold and heat leads to the dysfunction of the orifices, a disorder of the blood-governing organs leads to extravasation of blood", and advocated that the treatment should take into account the symptoms, syndromes, viscera, and seasons, and clarify the syndromes, which would provide a new research idea for the exploration of the TCM connotation of this disease.
3.Efficacy comparison of percutaneous vertebroplasty in the treatment of osteoporotic lumbar compression fracture via the approach through midpoint transverse process-transition zone of articular process and the unilateral transpedicular approach
Jiang WANG ; Jinpeng DU ; Shuai LI ; Xiaohui WANG ; Yunfei HUANG ; Mingzhe FENG ; Zechao QU ; Zhiyi TANG ; Baorong HE
Chinese Journal of Trauma 2023;39(5):427-434
Objective:To compare the efficacy of percutaneous vertebroplasty (PVP) in the treatment of osteoporotic lumbar compression fracture (OLCF) via the approach through midpoint transverse process-transition zone of articular process and the unilateral transpedicular approach.Methods:A prospective cohort study was conducted to analyze the clinical data of 794 patients with OLCF treated in Honghui Hospital affiliated to Xi′an Jiaotong University School of Medicine from January 2017 to December 2019. The patients were divided into transitional-zone puncture group (400 patients, 400 vertebrae) and pedicle puncture group (394 patients, 394 vertebrae) according to the envelope method. The transitional-zone puncture group was treated with PVP via the approach through midpoint transverse process-transition zone of articular process, and the pedicle puncture group was treated with PVP via the unilateral transpedicular approach. The operation time and radiation dose were documented. The visual analogue score (VAS) and Oswestry dysfunction index (ODI) were evaluated before operation and at 1 day, 3 months, 1 year after operation. The cement distribution and the incidence of complications such as cement leakage, re-fracture of the injured vertebra, spinal cord nerve injury and facet joint injury were detected.Results:The patients were composed of 270 males and 524 females, at the age of 68.9-78.5 years [(73.7±4.8)years]. All patients were followed up for 12-14 months [(13.4±0.8)months]. The operation time and radiation dose in transitional-zone puncture group were reduced compared with pedicle puncture group ( P<0.01). There was no significant difference in VAS and ODI between the two groups before operation (all P>0.05). The VAS between transitional-zone puncture group [(2.1±0.9)points, (2.3±1.1)points, (2.7±1.3)points] and pedicle puncture group [(2.3±0.7)points, (2.5±0.9)points, (2.9±1.1)points] was obviously reduced from that before operation (all P<0.01), significantly different at 1 day, 3 months, 1 year after operation ( P<0.05 or 0.01). The ODI between transitional-zone puncture group (14.3±1.8, 13.6±3.4, 11.3±4.4) and pedicle puncture group (25.5±5.7, 20.7±6.3, 20.6±6.9) was significantly different at 1 day, 3 months, 1 year after operation (all P<0.01), and all were obviously reduced from that before operation (all P<0.01). With regard to the cement distribution, the number of bilaterally cemented vertebrae in transitional-zone puncture group (324) was more than that in pedicle puncture group (94) ( P<0.01). The incidence of cement leakage, re-fracture of the injured vertebra and facet joint injury was 8.25%(22/400), 0.00%(0/400) and 3.25%(13/400) in transitional-zone puncture group, significantly different from 20.81%(82/394), 2.03%(8/394) and 9.90%(39/394) in pedicle puncture group ( P<0.05 or 0.01). There was no significant difference in spinal cord nerve injury between the two groups ( P>0.05). Conclusion:For OLCF, PVP via the approach through the midpoint transverse process-transition zone of articular process and the unilateral transpedicular approach are both effective, but the former has advantages of shorter operation time, smaller radiation dose, greater dispersion of bone cement and lower incidence of cement leakage, re-fracture of the injured vertebra and facet joint injury.
4.Efficacy of O-arm combined with CT three-dimensional navigation system assisted versus manual screw placement in the treatment of lower cervical fracture and dislocation
Shuai LI ; Jinpeng DU ; Jiang WANG ; Yunfei HUANG ; Zhigang ZHAO ; Zhen CHANG ; Xuefang ZHANG ; Liang YAN ; Hua HUI ; Xiaobin YANG ; Zhongkai LIU ; Lingbo KONG ; Bolong ZHENG ; Baorong HE
Chinese Journal of Trauma 2023;39(8):712-720
Objective:To compare the clinical efficacies of O-arm combined with CT three-dimensional navigation system assisted screw placement versus manual screw placement in treating lower cervical fracture and dislocation.Methods:A retrospective cohort study was used to analyze the clinical data of 41 patients with lower cervical fracture and dislocation, who were treated in Honghui Hospital, Xi′an Jiaotong University from May 2021 to February 2022. The patients included 26 males and 15 females, aged 31.5-48.6 years [(41.5±15.0)years]. The injured segments were C 3 in 3 patients, C 4 in 12, C 5 in 13, C 6 in 10 and C 7 in 3. Nineteen patients were treated with cervical pedicle screws by O-shaped arm combined with CT three-dimensional navigation system (navigation group, 76 screws) and 22 by bare hands (traditional group, 88 screws). The total operation time, effective operation time, single nail placement time, single screw correction times, screw distance from anterior cortex, intraoperative blood loss, intraoperative fluoroscopic radiation dose, incision length and length of hospital stay were compared between the two groups, and the height of intervertebral space, Cobb angle, interbody slip distance and American Spinal injury Association (ASIA) grade were compared before operation and at 3 days after operation. Visual analogue score (VAS), Japanese Orthopedic Association (JOA) score, and neck dysfunction index (NDI) were evaluated before operation, at 3 days, 3 months after operation and at the last follow-up. Accuracy of screw placement and incidence of complications (adjacent facet joint invasion, infection, screw loosening) were detected as well. Results:All the patients were followed up for 11.1-13.9 months [(12.5±1.4)months]. The total operation time, intraoperative blood loss, intraoperative fluoroscopic radiation dose and incision length in the navigation group were more or longer than those in the traditional group (all P<0.05). The effective operation time, single nail placement time, single nail correction times and screw distance from anterior cortex in the navigation group were markedly less or smaller than those in the traditional group (all P<0.05). There was no significant difference in the length of hospital stay between the two groups ( P>0.05). There were significant improvements in the height of intervertebral space, Cobb angle and interbody slip distance between the two groups at 3 days after operation (all P<0.05). There was no significant difference in the height of intervertebral space, Cobb angle, interbody slip distance or ASIA grade between the two groups before operation or at 3 days after operation (all P>0.05). Compared with pre-operation, the VAS, JOA score and NDI were significantly improved in both groups at 3 days, 3 months after operation and at the last follow-up (all P<0.05), with further improvement with time. There was no significant difference in VAS between the two groups before operation or at 3 months after operation (all P>0.05), but it was markedly lower in the navigation group compared with the traditional group at 3 days after operation and at the last follow-up (all P<0.05). There were no significant differences in JOA score or NDI between the two groups before operation or at 3 days and 3 months after operation (all P>0.05), but both were lower in the navigation group compared with the traditional group at the last follow-up (all P<0.05). The accuracies of placement of grade 0 and grade 0+1 screws were 92.0% (70/76) and 96.6% (73/76) in the navigation group, respectively, which were markedly higher than 88.7% (78/88) and 93.5% (82/88) in the traditional group (all P<0.05). The rates of adjacent facet joint invasion of A, B, and C degrees were 71.2% (54/76), 28.8% (22/76) and 0% (0/76) in the navigation group, respectively, while the invasion rates were 60.5% (53/88), 32.3% (28/88) and 7.3% (7/88) in the traditional group ( P<0.05). No screw loosening was noted in the navigation group, but the screw loosening rate was 9.1% (8/88) in the traditional group ( P<0.01). Conclusion:Compared with manual screw placement, O-arm combined with CT three-dimensional navigation system assisted screw placement for lower cervical fracture and dislocation has the advantages of shorter effective operation time, quicker screw placement, stronger screw holding force, better cervical stability, slighter postoperative pain, higher screw placement accuracy, and lower facet joint invasion and screw loosening rates.
5.Altered fear engram encoding underlying con-ditioned versus unconditioned stimulus initi-ated memory updating
Shuaiwen TENG ; Xinrong WANG ; Bowen DU ; Xiaolin CHEN ; Guanzhou FU ; Yunfei LIU ; Shuqi XU ; Jiachen SHUAI ; Zheyu CHEN
Chinese Journal of Pharmacology and Toxicology 2023;37(z1):64-64
OBJECTIVE It's known that post-retrieval extinction but not extinction alone could erase fear memory.However,whether the coding pattern of original fear engrams is remod-eled or inhibited remains largely unclear.Here we try to investigate whether the coding pattern of memory engrams is altered during post-retrieval extinction induced memory updating.METHODS To answer the question,by using activity-depen-dent neuronal-tagging technology,neuronal trac-ing technique combined with optogenetic manipu-lation and in vivo calcium imaging,we identified the fear and extinction cells in PrL and BLA and investigated the dynamic encoding of memory engram ensembles in the PrL and BLA during CS versus US initiated memory updating.RESULTS We found increased reactivation of engram cells in the prelimbic cortex and basolat-eral amygdala during memory updating.More-over,conditioned stimulus and unconditioned stimulu sinitiated memory updating depend on the engram cells reactivation in the prelimbic cor-tex or basolateral amygdala respectively.Finally,we found memory updating causes increased overlapping between fear and extinction cells and the original fear engrams encoding was altered during memory updating.CONCLUSION Our data provide the first evidence to show the overlapping ensembles between fear and extinc-tion cells and functional reorganization of original engrams underlying conditioned stimulus and unconditioned stimulus initiated memory updating.
6.Comparison of the efficacy of cervical decompression performed at different times in the treatment of incomplete cervical spinal cord injury
Shuai LI ; Yuan HE ; Yanzheng GAO ; Dianming JIANG ; Jun SHU ; Jian CHEN ; Jinpeng DU ; Lei ZHU ; Yunfei HUANG ; Zhen CHANG ; Liang YAN ; Hua HUI ; Xiaobin YANG ; Lingbo KONG ; Baorong HE
Chinese Journal of Trauma 2023;39(12):1070-1078
Objective:To compare the efficacy of cervical decompression performed at different times in the treatment of incomplete cervical spinal cord injury.Methods:A multicenter retrospective cohort study was conducted to analyze the clinical data of 96 patients with incomplete cervical spinal cord injury admitted to six hospitals including Honghui Hospital affiliated to Xi'an Jiaotong University, etc, from May 2018 to May 2021. There were 36 females and 60 males, aged 28-42 years [(35.2±6.7)years]. The injured segments were at C 3 in 7 patients, C 4 in 15, C 5 in 20, C 6 in 23 and C 7 in 31. According to the American Spinal Injury Association (ASIA) scale, there were 59 patients with grade B, 27 grade C, and 10 grade D. A total of 36 patients underwent cervical decompression within 24 hours after injury (early group), 33 patients within 24-72 hours after injury (late group), and 27 patients within 4-14 days after injury (delayed group). The operation time, intraoperative blood loss, postoperative drainage, length of hospital stay, Cobb angle, height of intervertebral space and space occupation of the spinal canal before surgery and at postoperative 3 days, and ASIA score, ASIA motor score, ASIA light tactile score, ASIA acupuncture sensation score, visual analog scale (VAS) score, Japanese Orthopedic Association (JOA) score, neck dysfunction index (NDI) before surgery and at postoperative 3 months, 1 year and at the last follow-up and incidence of complications were compared among the three groups. Results:All the patients were followed up for 12-21 months [(16.4±4.2)months]. There was no significant difference in the operation time among the three groups (all P>0.05). The intraoperative blood loss and postoperative drainage volume in the early group were (312.5±5.2)ml and (165.3±45.8)ml, which were higher than those in the late group [(253.5±40.0)ml, (120.4±60.6)ml] and the delayed group [(267.3±36.8)ml and (130.4±38.6)ml] (all P<0.01). There was no significant difference between the late group and the delayed group (all P>0.05). The length of hospital stay in the early group was (5.2±1.6)days, which was shorter than that in the late group [(7.6±2.3)days] and the delayed group [(8.0±1.3)days] (all P<0.05), but there was no significant difference between the late group and the delayed group ( P>0.05). There was no significant difference in the Cobb angle, height of intervertebral space and space occupation of the spinal canal among the three groups before and at postoperative 3 days (all P>0.05). There was no significant difference in the ASIA score, ASIA motor score, ASIA light tactile score, ASIA acupuncture sensation score, VAS score, JOA score and NDI among the three groups before surgery (all P>0.05). At postoperative 3 months, 1 year and at the last follow-up, the ASIA grading of the early group was better than that of the late group and the delayed group ( P<0.05 or 0.01), but there was no statistically significant difference between the late group and the delayed group (all P>0.05). The ASIA motor scores of the early group were (56.4±4.5)points, (76.3±3.6)points and (85.4±6.5)points at postoperative 3 months, postoperative 1 year and the last follow-up, respectively, which were higher than those in the late group [(52.3±2.4)points, (60.3±8.6)points and (72.3±2.4)points] and the delayed group [(51.9±2.3)points, (62.8±4.6)points and (71.9±1.3)points]; the ASIA light tactile scores of the early group were (70.2±2.9)points, (72.6±4.3)points and (78.3±2.3)points, which were higher than those in the late group [(66.2±3.7)points, (68.3±1.6)points and (73.3±1.6)points] and the delayed group [(65.2±2.1)points, (67.8±1.9)points and (72.3±2.5)points]; acupuncture sensation scores of the early group were (71.9±3.1)points, (80.1±3.8)points and (89.1±7.6)points, which were higher than those in the late group [(67.4±2.7)points, (72.6±3.7)points and (77.9±1.8)points] and the delayed group [(68.3±2.2)points, (72.6±3.1)points and (77.2±1.9)points] (all P<0.05). VAS scores of the early group at postoperative 3 months, 1 year and at the last follow-up were (4.3±0.6)points, (2.4±0.3)points and (1.6±0.2)points, which were lower than those in the late group [(5.1±1.3)points, (4.1±0.6)points and (3.0±0.6)points] and the delayed group [(5.0±1.7)points, (4.0±0.8)points and (3.1±0.2)points]; JOA scores of the early group were (12.8±1.6)points, (14.4±2.6)points and (17.9±3.3)points, which were higher than those in the late group [(11.9±1.9)points, (13.3±1.6)points and (8.9±1.3)points] and the delayed group [(11.6±1.8)points, (13.2±1.4)points and (9.3±2.1)points]; NDI scores of the early group were 12.1±3.3, 10.1±2.1 and 7.3±1.4, which were lower than those in the late group (14.4±3.1, 12.3±1.6 and 8.9±1.3) and the delayed group (14.1±2.3, 12.9±1.9 and 9.5±2.1) (all P<0.05). There was no significant difference in all the above-mentioned scores at postoperative 3 months, 1 year and at the last follow-up between the late group and the delayed group (all P>0.05). The incidence of complications was 25.0% (9/36) in the early group, 27.3% (9/33) in the late group and 37.0% (10/27) in the delayed group (all P>0.05). Conclusion:Compared with within 24-72 hours and 4-14 days after injury, cervical decompression performed within 24 hours after injury for patients with incomplete cervical spinal cord injury can shorten the length of hospital stay, improve the function of the spinal cord nerves and relieve pain, with no increase of the incidence of complications.
7.miRNA-6516-5p regulates the proliferation and migration of renal cancer cells by targeting ODC1
Geng HUANG ; Dingwen GUI ; Zuwei XU ; Jinlun FU ; Shuai LUO ; Yunfei ZHAO ; Jinghua WAN
International Journal of Surgery 2022;49(3):194-198,C3
Objective:To explore the expression of microRNA (miRNA)-6516-5p in renal cancer cell lines and the molecular mechanisms regulating the proliferation and migration of renal cancer cells.Methods:quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-6516-5p in renal cancer cell lines and normal proximal renal tubular epithelial cell lines. The liposome method was used to transiently transfect miR-6516-5p mimic and nonsense sequence (NC) into renal cancer cells with the lowest expression of miR-6516-5p, namely miR-6516-5p group and NC group. qRT-PCR was used to detect the expression of miR-6516-5p in transfected cells. CCK-8 and Transwell migration experiment were used to detect the proliferation and migration of transfected cells. Bioinformatics software and dual luciferase gene report experiment were used to predict and verify the regulation of miR-6516-5p on target gene, respectively. qRT-PCR and Western blotting were used to detect the expression of target gene in transfected cells. Measurement data were expressed as mean±standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The expression of miR-6516-5p in renal cancer cell lines was significantly lower than that of normal proximal tubular epithelial cells ( P<0.01), and the expression of miR-6516-5p in 786-O cells was the lowest ( F=27.69, P<0.01). The expression of miR-6516-5p in 786-O cells in NC group and miR-6516-5p group was 1.01±0.08 and 9.91±1.16, respectively. Compared with the NC group, the expression of miR-6516-5p in 786-O cells in the miR-6516-5p group was significantly increased ( t=7.63, P<0.01). Up-regulation of miR-6516-5p can significantly inhibit the proliferation of 786-O cells ( P<0.05). The migration numbers of NC group and miR-6516-5p group were 85.65±8.77 and 28.05±6.20, respectively. Overexpression of miR-6516-5p could inhibit the migration of 786-O cells ( t=5.36, P< 0.01). The target gene of miR-6516-5p may be ornithine decarboxylase 1 ( ODC1), miR-6516-5p can significantly inhibit the luciferase activity of wild-type ODC1-3′UTR ( t=9.83, P<0.01). Up-regulation of miR-6516-5p can reduce the expression of ODC1 mRNA and protein in 786-O cells ( P<0.01). Conclusion:The expression of miR-6516-5p is reduced in renal cancer cell lines, miR-6516-5p inhibits the proliferation and migration of renal cancer 786-O cells by targeting ODC1, miR-6516-5p may become a potential molecular target of renal cancer.
8.Physcion regulates the cell cycle and proliferation of prostate cancer through miR-380-3p
Zuwei XU ; Dingwen GUI ; Jinlun FU ; Shuai LUO ; Yunfei ZHAO ; Geng HUANG ; Jinghua WAN
International Journal of Surgery 2022;49(3):198-202,C4
Objective:To investigate the mechanism of physcion affecting the cell cycle and proliferation of prostate cancer DU145 cell line by regulating the expression of miR-380-3p.Methods:Prostate cancer DU145 cells were treated with 50 μg/mL physcion as physcion group, and normal cultured DU145 cells without any treatment were used as control group. Flow cytometry was used to detect DU145 cell cycle changes. MTT proliferation test was used to detect the proliferation of DU145 cells. quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-380-3p in DU145 cells. The bioinformatics software RNAhybrid was used to predict the target genes of miR-380-3p. qRT-PCR and Western blotting methods were used to detect the expression of miR-380-3p target gene. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups. Results:Compared with the control group, DU145 cells in the physcion group were blocked in the G 0/G 1 phase ( P<0.01), and the proliferation ability of DU145 cells was significantly inhibited ( P<0.05). The expression of miR-380-3p in DU145 cells in the control group and physcion group was 8.36 ± 1.42 and 1.08 ± 0.39, respectively. Physcion could promote the expression of miR-380-3p ( t=4.96, P<0.01). The functional target gene of miR-380-3p may be UHRF1. The relative expression levels of UHRF1 mRNA in DU145 cells in the physcion group and control group were 0.23±0.06 and 1.04±0.15, respectively. Compared with the control group, the expression of UHRF1 gene in DU145 cells in the physcion group was decreased ( t=4.55, P<0.01). Conclusion:Physcion can inhibit the proliferation of prostate cancer DU145 cells and induce G 0/G 1 block in DU145 cells, which may be closely related to the regulation of miR-380-3p.
9.lncRNA AC068768.1 regulates the cycle and proliferation of renal cancer cells by targeting miR-21-5p
Zuwei XU ; Dingwen GUI ; Jinlun FU ; Shuai LUO ; Yunfei ZHAO ; Geng HUANG ; Jinghua WAN
International Journal of Surgery 2021;48(6):387-391,F4
Objective:To explore the effect of long non-coding RNA (lncRNA) AC068768.1 on the cycle and proliferation of renal cancer cells and its molecular mechanism.Methods:Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression of AC068768.1 in renal cancer cell lines. The OS-RC-2 cells with the lowest expression of AC068768.1 were used as the transfection objects, OS-RC-2 transfected with the negative control plasmid was set as the control group, and the cells transfected with the AC068768.1 plasmid were set as the AC068768.1 group. qPCR was used to detect the expression of AC068768.1 in transfected OS-RC-2 cells. The effects of AC068768.1 on the cell cycle and proliferation of OS-RC-2 were detected by flow cytometry and tetramethylazazole blue colorimetric (MTT) proliferation experiments. Using bioinformatics methods to predict the microRNA (miRNA) that AC068768.1 may bind. qPCR was used to detect the expression of miRNA and downstream gene mRNA, and Western blot was used to detect the expression of downstream gene protein.The measurement data were expressed as mean±standard deviation ( Mean± SD), the comparison between the two groups adopts the t-test, and the comparison among multiple groups adopts the One-way analysis of variance. Results:Compared with normal renal tubular epithelial cells, the expression of AC068768.1 in renal cancer cell lines was significantly reduced, the difference was statistically significant ( P<0.01). The expression of AC068768.1 in OS-RC-2 cells in the AC068768.1 group was significantly higher than that in the control group, the difference was statistically significant ( P<0.01). Up-regulating the expression of AC068768.1 can inhibit the cycle ( P<0.05) and proliferating ability ( P<0.05) of renal cancer cells. miR-21-5p may be the functional target gene of AC068768.1. Up-regulation of AC068768.1 can significantly inhibit the expression of miR-21-5p ( P<0.01) and promote the expression of tissue inhibitor of metalloproteinase 3 (TIMP3) ( P<0.01). Conclusion:AC068768.1 promotes the expression of TIMP3 gene by regulating the expression of miR-21-5p, thereby inhibiting the cell cycle and proliferation of renal cancer OS-RC-2 cells.
10.Rapid cryopreservation for Siraitia grosvenorii cells based on cells' capacitance detection.
Jiarui LI ; Zejian WANG ; Meijin GUO ; Yuanxin GUO ; Shuai HUANG ; Yunfei SONG ; Zhen SUN ; Yangyang SUN ; Fanjing KONG ; Yingping ZHUANG
Chinese Journal of Biotechnology 2017;33(5):817-827
A rapid quantitative evaluation method for Siraitia grosvenorii cells was successfully developed based on plant cells' capacitance value detected by a viable cell mass monitor and the cryopreservation of S. grosvenorii suspension cells was optimized. The survival rate of S. grosvenorii cells was quantitatively measured by viable cell mass monitor and 2, 3, 5-triphenyltetrazolium chloride (TTC). An optimum cryoprotectant recipe is that the growth medium contained 10% sucrose and 10% DMSO. The experimental results also showed higher cell survival rates and cell viabilities were achieved when suspension cells were treated with pretreatment of 0.2 mol/L sucrose. With the increase of concentration of sucrose, however, the cell survival rate was decreased. And the cell survival rate represented a bell shape with the increase of pretreatment time. The highest cell survival rate and cell viability were obtained with the 9 h' s pretreatment. In addition, there was a good correlation between the cell survival rate measured by cell recovery test and that measured by viable cell mass monitor, while there were no significant differences in the cell morphology and the ability of mogrosides V production by S. grosvenorii cells cultured in suspension after cryopreservation. Therefore, the evaluation method developed based on the viable cell mass monitor has good feasibility and reliability.

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