1.Bone morphogenetic protein-2 gene affects the histology and collagen type Ⅰ andⅡ expressions in degenerative intervertebral disc
Chinese Journal of Tissue Engineering Research 2013;(43):7501-7506
BACKGROUND:Studies have shown that the changes of extracellular matrix in degenerative intervertebral disc tissues mainly present as the decrease of col agen type Ⅱ and proteoglycan contents and the increase of col agen type Ⅰ content. OBJECTIVE:To explore the effect of adeno-associated virus-mediated bone morphogenetic protein 2 gene on nucleus pulposus Ⅰ and Ⅱ col agen levels in rabbit degenerative intervertebral disc tissues. METHODS:L 2-3 , L 3-4 , L 4-5 and L 5-6 lumbar discs of 12 New Zealand white rabbits were punctured to establish interverbral disc degeneration model. Subsequently, 12 rabbits were randomly divided into three groups, four rabbits in each group. The intervertebral discs in the adeno-associated virus-mediated bone morphogenetic protein 2 group were injected with the adeno-associated virus-mediated bone morphogenetic protein 2 gene, the intervertebral discs in the adeno-associated virus group were injected with adeno-associated virus only, while the discs in the normal saline group were injected with normal saline. Al rabbits were sacrificed after injected for 8 weeks, and the L 2-3 , L 3-4 , L 4-5 and L 5-6 lumbar discs of each rabbit were col ected, paraffin-embedded and sliced. The histological changes of nucleus pulposus were observed with hematoxylin-eosin staining, and the immunohistochemistry was used to detect the col agen type Ⅰ and Ⅱ expressions in nucleus pulposus. Semi-quantitative analysis was performed. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that the nucleus pulposus in the intervertebral disc tissues was less in the adeno-associated virus-mediated bone morphogenetic protein 2 group, the nucleus pulposus was in single or clustered distribution with clear nucleus structure and without fibrous tissue fil ing. The tissue structures of nucleus pulposus were the same in the adeno-associated virus group and normal saline group, the cellnumber in nucleus pulposus was smal , the nucleus pulposus was shrunken and shriveled, and the cells were fil ed with fibrous tissue and arranged disorderly. Immunohistochemistry staining showed the expression of col age type Ⅰ in the intervertebral disc nucleus pulposus of adeno-associated virus-mediated bone morphogenetic protein 2 group was higher than that of the adeno-associated virus group and normal saline group (P<0.05);the expression of col age typeⅠ in the intervertebral disc nucleus pulposus of adeno-associated virus-mediated bone morphogenetic protein 2 group was lower than that of the adeno-associated virus group and normal saline group (P<0.05). The results indicate that adeno-associated virus-mediated bone morphogenetic protein 2 can inhibit the expression of col agen type Ⅰ in the intervertebral disc nucleus pulposus, promote the expression of col agen type Ⅱ. Maintaining the content of col agen in intervertebral disc can keep the histological structure and morphology of intervertebral disc, stabilize the environment for nucleus pulposus cellgrowth, and delay the intervertebral disc degeneration.
2.Anatomy study and clinical applications of complex tissue flap pedicled with inferior gluteal artery perforator for repairing giant sacrococcygeal pressure sore
Yun XIE ; Jianhua LIN ; Junjian YE ; Heping ZHENG
Chinese Journal of Microsurgery 2014;37(4):373-376
Objective To discuss the technical feasibility and clinical effectiveness of using complex tissue flap pedicled with inferior gluteal artery perforator for repair giant sacrococcygeal pressure sore.Methods Thirty embalmed lower limbs of adult cadavers perfused with red latex were used for anatomical study,and the followings were observed:①The course,branche and distribution of gluteal artery.②The course and distribution of the posterior femoral cutaneous nerve.③Anastomosis between the posterior cutaneous branch of gluteal artery and nutrient vessels of the posterior femoral cutaneous nerve.8 cases aging from 17 years to 56 years were completed during May 2007 to July 2013,6 cases were males and 2 cases were females.The sizes of pressure sore with the depth to Ⅳ degree were ranged from 16 cm × 9 cm to 22 cm × 10 cm.The sizes of flaps were harvested from 32 cm × 10 cm to 25 cm × 9 cm.Results The gluteal artery crossed the edge of the piriformis,the main stem was (3.1 ± 0.4) mm in diameter and gave out 2-5 muscular branches to supply the gluteus maximus.The posterior femoral cutaneous nerve crossed the edge of gluteus maximus and descended between biceps femoris and semitendinosus.Perforating deep fascia point located was (5.9 ± 0.8) cm above the line between medial and lateral femoral epicondyle.The constant anastomosis were formed by the posterior cutaneous branch of gluteal artery,the obturator artery perforator and the direct popliteal artery perforator around the posterior femoral cutaneous nerve.The complex flap survived successfully in all patients.Sutures were removed at 14 days postoperatively and the wounds healed well.All supplied areas were closed by directly suturing.Recurrent sacrococcygeal pressure sore was not observed in all cases with satisfied appearance and normal color during the outpatient follow-up period from 5 months to 5 years.Conclusion The united flap of gluteal myocutaneous flap and the posterior femoral cutaneous neurovascular flap pedicled with inferior gluteal artery perforator can be used to primary repair giant sacrococcygeal pressure sore.Rich blood supply,simple operation technique and high rate survival rate was considered as advantages of the flap.The lower recurrence of pressure sore was due to nice wear resisting with rich layer of anatomical structure in the flap and strong ability of anti-infection.The clinical effect was satisfied.
3.Effect of hypothyroidism on c-fos and c-jun mRNA expression in testes of rats
Juwei WANG ; Zhonghui YE ; Panhong GOU ; Junjian MA ; Yanling WANG ; Junling WANG
Chinese Journal of Endemiology 2016;(1):27-31
Objective This study aims to define the effects of hypothyroidism on c-fos and c-jun mRNA expression in rat testes to provide a theoretical basis for prevention and control of iodine deficiency disorders (IDD). Methods According to body weight (200 - 240 g), 20 Wistar male rats were divided into control group and hypothyroidism group (1 ml/100 g, 0.1% propylthiouracil by intragastric administration) by digital table. There were 10 male rats in each group and body weight was observed every 3 days. After 60 days, all rats were killed. The levels of thyroid hormones [total triiodothyronine (TT3), total thyroxine (TT4), and thyroid stimulating hormone (TSH)] were measured by radioimmunoassay. The mRNA expression levels of c-fos and c-jun in testes were detected by real-time quantitative PCR. Results Compared with control groups [(298.20 ± 12.15), (344.00 ± 13.73) g], the weights of hypothyroidism groups in 30 days [(239.00 ± 15.02) g] and in 60 days [(232.67 ± 17.86) g] were significant decreased (t=7.704, 11.380, all P<0.05). The levels of TT3 [(373.32 ± 101.31) ng/L] and TT4 [(4.00 ± 0.89) × 103 ng/L] in serum of hypothyroidism group were found to be significantly decreased, whereas the level of TSH [(5.77 ± 0.89) × 103 U/L] was increased in comparison with those of the control groups [(1 000.01 ± 273.53) ng/L, (44.33 ±7.84) × 103 ng/L, (1.87 ± 0.70) × 103 U/L, t = 5.262, 12.520, 8.413, all P< 0.05]. Compared with control group (1.00 ± 0.08, 1.01 ± 0.04), the c-fos and c-jun mRNA expression (0.67 ± 0.03, 0.75 ± 0.02) of hypothyroidism group was significant decreased (t = 12.382, 13.784, all P < 0.05). Conclusion Hypothyroidism may reduce mRNA expression levels of testicular c-fos and c-jun, and then damage the reproductive system in male rats.
4.Expression of miR-17-92 cluster in osteosarcoma and its clinical significance
Jinluan LIN ; Fasheng WANG ; Junjian YE ; Zhaoyang WU ; Xiang LI ; Jianhua LIN
Chinese Journal of Clinical Oncology 2014;(23):1532-1535
Objective:To determine the expression of miR-17-92 cluster in osteosarcoma tissue samples and explore its associa-tion with clinical significance. Methods: Quantitative polymerase chain reactiom analysis was used to examine the expression of miR-17-92 cluster in osteosarcoma tissues. Normal bone tissues from 63 patients were matched, and the relationships between the ex-pression of miR-17-92 cluster and the clinicopathological features and prognosis of osteosarcoma were explored. Results:The relative expression of miR-17-92 cluster in osteosarcoma tissues was significantly higher than those in adjacent normal tissues (P<0.05). The high expression of miR-17-92 had a significant correlation with reduced survival (P=0.027). Conclusion:The expression of miR-17-92 cluster closely correlates with the occurrence and progress of osteosarcoma and may be used as an indicator for osteosarcoma prognosis.
5.Cloning and Expression of Various Deletants of Gsalpha Gene in Escherichia Coli
Jinxing LOU ; ; Junjian HUANG ; Yiling SI ; Jiezhi LI ; Yali ZHAO ; Qinong YE ; Cuifen HUANG
Journal of Experimental Hematology 2000;8(4):251-254
Gsalpha gene mutation has been discovered in some human tumors. In our previous studies, three novel deletants of Gsalpha gene, Gsalpha L-1(500 bp), Gsalpha L-2(300 bp), and Gsalpha L-3(200 bp), and wild type Gsalpha-4(1 200 bp) were found in human leukemia cell lines and detected in leukemic cells from patients with acute leukemia. To investigate the construction, function and biological significance of the deletants, the plasmids of Gsalpha L-1, Gsalpha L-2 and wild Gsalpha-4 were transformed into E. coli DH5, amplified by PCR, and cloned in expression vector pET22b(+), and then transformed into E. coli, respectively. As a result, higher levels of expression of three recombinants were obtained in form of inclusion bodies. The results suggested that these Gsalpha isoforms have an open reading frame of gene and can be expressed in vitro. The data lay a foundation to study the relation of Gsalpha gene to leukemogenesis.