1.In vitro isolation and culture of primary blood-brain barrier endothelial cells by double filtering technique
Jiexiao LIU ; Chuanqiang PU ; Yanlei HAO
Chinese Journal of Tissue Engineering Research 2007;0(21):-
96% 7-10 days after culture. Immunohistochemistry of Ⅷ factor polyclone antibody showed positive in cytomembrane and cytoplasm, but negative in cell nucleus. These were cells with blood-brain barrier. CONCLUSION: Ideal endothelial cells of blood-brain barrier can be cultured by the improved cultural method of double filtering.
2.Treatment of elderly severe osteoporotic vertebral compressive fractures by kyphoplasty
Hai TANG ; Hao CHEN ; Bingqiang WANG ; Jinjun LI ; Pu JIA
Chinese Journal of Orthopaedics 2010;30(10):978-983
Objective To evaluate the feasibility and therapeutic effect of kyphoplasty in treating severe osteoporotic vertebral compressive fractures. Methods Thirty-five patients (48 vertebral bodies) with severe osteoporotic compressive fractures were included. There were 33 females and 2 males with the mean age of 74.2 years. The average compressive rate of the affected vertebral bodies was 77.0%. The thoracolumbar vertebrae were treated with kyphoplasties. Percutaneous puncture direction was adjusted according to compressive rate and shape of the vertebral bodies. The inflatable bone tamp was inserted into the fractured vertebral body. The balloon was inflated with low pressure and dilate-relieve-dilate method was applied. The balloon was deflated and withdrawn, leaving a cavity within the vertebral body, which then fulfilled with visualized bone cement. Preoperative and postoperative symptom level, complications and radiographic findings were recorded. Results All 35 patients tolerated procedure well. The mean heights of the anterior, mid and posterior vertebral body had improved from (0.8±0.1) cm, (0.8±0.2) cm, (2.1 ±0.8) cm preoperatively to (1.2±0.3) cm, (1.3±0.2) cm, (2.3±1.0) cm respectively after operation (P <0.05). There was significance difference between preoperative and postoperative heights of the anterior and mid vertebral body. The mean kyphosis was improved from 28.2°±5.2° before operation to 19.1°±4.9° after operation. Conclusion Kyphoplasty is feasible and effective for severe osteoporotic vertebral compressive fractures.
4.Endoplasmic reticulum stress participates in uric acid-induced phenotypic transformation of renal tubular epithelial cells
Pu WU ; Fei ZHAO ; Dan NIU ; Xinyang WANG ; Yaning HAO
Chinese Journal of Nephrology 2016;32(12):922-927
Objective To explore the effect of endoplasmic reticulum stress (ER stress) in uric acid?induced phenotypic change in renal tubular epithelial cells (HK?2). Methods (1) HK?2 cells were cultured with 0, 75, 150, 225, 300 mg/L uric acid for 24 h in vitro. (2) The cells were divided into normal control group, ER stress inhibitor 4?PBA (5 μmol/L) group, uric acid (150 mg/L) group and 4?PBA+uric acid group for 24 h. Morphological changes of HK?2 cells were observed under inverted microscope. MTT assay was used to detect the proliferation of HK?2 cells treated with 150 mg/L uric acid for 24, 48 and 72 h. The protein expressions of α?smooth muscle actin (α?SMA), vimentin, snail, glucose regulated protein 78 (GRP78) and the phosphorylation of eukaryotic initiation factor 2α(p?eIF2α) in HK?2 cells were measured by Western blotting. Results Compared with the control group, HK?2 cells in uric acid groups (150, 225, 300 mg/L) showed fibroblast?like appearance. The protein expressions of α?SMA, vimentin, snail, GRP78 and p?eIF2α in 150 mg/L and 225 mg/L uric acid groups were higher than those in the control group (all P<0.05). The proliferation of HK?2 cells in 150 mg/L uric acid group was lower than that in control group at 48 and 72 h (all P<0.01). Compared with the uric acid group, the cell morphology in 4?PBA+uric acid group was improved, and the protein expressions ofα?SMA, vimentin, snail, GRP78 and p?eIF2α were decreased (all P<0.05). Conclusions Uric acid may induce the phenotype transformation of renal tubular epithelial cell, and ER stress is involved. 4?PBA may inhibit the uric acid?induced ER stress response and phenotypic transformation, and may be beneficial in attenuating uric acid?induced renal tubular damage.
5.Effects of cisplatin combined with heparanase inhibitor on proliferation and invasion of human nasopharyngeal carcinoma cells.
Yang LI ; Hao LIU ; Yingying HUANG ; Longjian PU ; Xudong ZHANG ; Zhiwen JIANG ; Chenchen JIANG
Acta Pharmaceutica Sinica 2013;48(4):609-14
This study is to investigate the effects of cisplatin combined with heparanase inhibitor OGT2115 on proliferation, invasion and migration of human nasopharyngeal carcinoma cells CNE-2 and to provide a new target for the treatment of metastasis of nasopharyngeal carcinoma. MTT assay was used to detect the cell viability of CNE-2 after exposure to different concentrations of DDP (2, 4, 8, 16 and 32 micromol x L(-1)), different concentrations of OGT2115 (0.4, 0.8, 1.6, 3.2 and 6.4 micromol x L(-1)), and DDP combined with OGT2115. Transwell assay was applied to analyze the effects of drugs on invasion and migration of human nasopharyngeal carcinoma cells. Wound healing assay was performed to detect cell migration and heparanase activity was analyzed by ELISA. MTT results showed that DDP can inhibit the proliferation of CNE-2 cells in a time and concentration-dependent manner, with an IC50 of 24.03 micromol x L(-1) at 24 h (P < 0.05), low concentration of DDP has almost no inhibitory effect on cell invasion and migration. DDP combined with OGT2115 can significantly inhibit cell invasion and migration. Inhibition of heparanase can significantly enhance anti-invasion and anti-proliferation of DDP.
6.Regulation of miR-19a and miR-19b on tumor suppressor gene-SEPT7
Kun WANG ; Guangxiu WANG ; Peiyu PU ; Anling ZHANG ; Jianwei HAO ; Zhifan JIA
Journal of International Oncology 2011;38(3):233-236
objective To identify SEPT7as one of the target genes of miR-19a and miR-19b.MethodsmiR-19a inhibitor and miR-19b inhibitor mediated by lipofectamine2000,were transfected to SNB19 glioma cells for knocking down miR-19a/19b overexpression.Real time PCR was conducted to detect the expression of miR-19a/miR-19b in transfected cells.The expression of SEPT7was determined by Western blot analysis.RT-PCR was used to detect the mRNA expression of SEPT7; and Luciferase reporter assay was used to identify the direct regulation of miR-19a/19b on SEPT7.ResultsIn SNB19 glioma cells transfected with miR-19a/19b inhibitor,the expression of miR-19a/miR-19b was significantly reduced,whereas the protein expression of SEPT7 was upreguhtted; no significant change of SEPT7 mRNA level was found and luciferase activity became stronger as compared to control cells.ConclusionSEPT7 is the target negatively regulated by miR-19a and miR-19b.
7.Clinical features and outcomes of seronegative rheumatoid arthritis in western China after one-year treatment with traditional disease modifying anti-rheumatoid drugs
Jing LUO ; Yanhua WANG ; Bomiao JU ; Xiaohong Lü ; Dan PU ; Yining SUN ; Zhiming HAO ; Lan HE
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(1):88-91
ABSTRACT:Objective To investigate the clinical features of seronegative rheumatoid arthritis (RA)in western China and its outcomes after one-year treatment with disease modifying anti-rheumatoid drugs (DMARDs) so as to provide evidence for effective therapy.Methods We made a retrospective analysis of 240 RA patients treated in our department from May 2013 to June 2014.We compared the 47 seropositive and 25 seronegative RA patients in clinical features,laboratory parameters and outcomes after one-year DMARDs medication.Results The percentage of seronegative RA was 10.4% (25/240).The number of swollen small joints was significantly smaller in seronegative RA group (P<0 .0 1 ).Compared with those in seropositive RA,the level of hemoglobin was lower,the level of platelets was higher,and the level of alkaline phosphatase was lower in seronegative RA (P<0 .0 5 ).The remission rate was higher in seronegative RA group than in seropositive RA group after one-year DMARDs administration (P<0.05).Conclusion Seronegative RA is not rare in clinic.Even though seronegative RA patients often present fewer swollen small joints, it is difficult to distinguish between seronegative and seropositve RA just based on the clinical features.Besides,hematological damage is more severe in some patients with seronegative RA.Only after one-year treatment with DMARDs,the remission rate is higher in seronegative RA patients than in seropositve RA ones.
8.The clinical study of the spinous process fusion plate for lumbar disc herniation
Hai TANG ; Tidong ZHANG ; Hao CHEN ; Pu JIA ; Li BAO ; Fei FENG
Chinese Journal of Orthopaedics 2016;36(6):344-352
Objective To study the clinical effect of the BacFuse for the lumbar disc herniation(LDH) and to discuss the affect for symptom of patients and structure of spine.Methods Collected the patients suffered from LDH retrospectively who had been implanted the BacFuse.There were 25 patients (29 surgical levels) included in the study.The clinical and X-ray assessment have been done on preoperation,postoperation,3 month,6 month and 12 month.Clinical index used visual analogue scale (VAS) score,oswestry disability index (ODI) score and Japanese Orthopaedic Association (JOA) score assessed clinical symptom and physical function.The intervertebral angle,posterior disk height and foraminal height were measured in X-ray.The patients were taken CT at preoperation and postoperation.The area and distance of herniated disc were measured in CT scan.Results In the aspect of clinical effect,25 cases were finished operation successly.The VAS score decreased from (6.61±1.29) points on preoperation to (2.78±1.00) points on the last follow-up,the ODI score decreased from (31.9±8.3) points to (15.9±6.4) points,the JOA score increased from (9.6± 3.0) points to (18.8± 3.2) points.In the aspect of spine structure,the intervertebral angle decreased from 10.1°±5.4° on preoperation to 8.4°±4.9° on the last follow-up,the posterior disk height increased from (0.65±0.29) cm on preoperation to (0.78±0.27) cm on the last follow-up and the foraminal height increased from (2.01±0.35) cm on preoperation to (2.16±0.37)cm.There was significant difference on posterior disk height and foraminal height between preoperation and postoperation,while the posterior disk height lost (0.07±0.12) cm (9.3%) and foraminal height lost (0.06±0.19) cm (2.7%).The CT showed that area of herniated disc decreased from (4.10±0.78) mm to (3.72±0.66) mm (9.3%) and distance of herniated disc decreased from (96.4±37.8) mm2 to (83.8±31.1) mm2 (7.4%).Conclusion BacFuse can relieve symptom and affect the biomechanics of the spine,makes the herniated disc retract partially,playing a Internal Retractor role in spine,and it is a new effective therapy for LDH.
9.Breeding Actinobacillus succinogenes with Acid-tolerance by Genome Shuffling
Xuan LIU ; Pu ZHENG ; Ye NI ; Jin-Jun DONG ; Zhi-Hao SUN ;
Microbiology 2008;0(11):-
A strain Actinobacillus succinogenes CGMCC 1593 was selected as the parent strain.After UV-EMS and UV-DES treatments respectively,seven mutated strains with subtle improvements in acid tol-erance were obtained,and were subjected for recursive protoplast fusion.Through three rounds of genome shuffling,four shuffled strains with both higher yield and acid tolerance were obtained.The shuffled strain namely F3-21 could even survive at pH 5.2.The comparison of the shuffled strains and the parent strain for succinic acid production was also studied here.After 48 h of shake-flask fermentation,the succinic acid concentration of F3-21 was 48% higher than that of the parent strain.When F3-21 was carried out in a 5 liter stirred bioreactor with pH controlled 5.6~6.0,the accumulation of succinic acid in 48 h fermentation attained 38.1 g/L,which was increased by 45% compared with that of the parent strain(26.2 g/L).While pH was controlled at 6.5~7.0,the production of succinic acid in 32 h fermentation attained 40.7 g/L.When F3-21 was carried out in fed-batch fermentation,succinic acid concentration of 67.4 g/L was reached in 72 h fer-mentation.These results indicated that the genome shuffling could improve the acid tolerance and the suc-cinic acid production of A.succinogenes CGMCC 1593.
10.The therapeutic effects of Fuyanshu Capsule on chronic endometritis in rats
Pu HUANG ; Jun ZHOU ; Gang CUI ; Baode YANG ; Wenbao ZHAO ; Chuanhao LIU ; Hao HU
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(3):404-407,413
Objective To observe the effects of Fuyanshu Capsule on phenol mucilage-induced endometritis rats and the possible anti-inflammation mechanism of the therapeutic effects.Methods Chronic endometritis in rats was induced by injection of phenol mucilage suspension into the uterus.Sixty female SD rats were randomly divided into 6 groups,namely,sham-operation group (distilled water,10mL/kg),model group,Jinji capsule group (0.65 g/kg),and Fuyanshu Capsule groups (1.8 g/kg,0.9 g/kg and 0.45 g/kg).After the rats were treated 28 days with corresponding medicine by intragastric administration,the pathology of the endometrium and changes of tumor necrosis factor α(TNF-α)and interleukin-2 (IL-2)levels were detected to evaluate the effects of Fuyanshu Capsule. Results Fuyanshu Capsule (1.8 g/kg and 0.9 g/kg)ameliorated the body weight reduction caused by endometritis in rats.Fuyanshu Capsule (1.8 g/kg,0.9 g/kg and 0.45 g/kg)reduced the ratio of the swelling uterus and ovaries to body weight of the rats.It ameliorated obviously the hyperplasia,necrosis and degeneration of endometrial epithelia and infiltration of inflammatory cells.The capsule (1.8 g/kg)decreased the serum IL-2 level in the rats with phenol mucilage-induced endometritis. Conclusion The anti-inflammatory effect of Fuyanshu Capsule on chronic endometritis induced by phenol mucilage in rats may be related to the decrease of IL-2 level.