1.Effects of Yaotu Granules on nuclear factor-kappaB signaling pathway in human nucleus pulposus cells
Shenghua HE ; Weiwei TAN ; Zhitao SUN ; Yeguang WANG ; Jian WANG ; Juyi LAI
Chinese Journal of Tissue Engineering Research 2016;20(33):4933-4939
BACKGROUND:Nuclear factor-κB (NF-κB) is highly expressed in the degenerated intervertebral disc. The Herbal Compound formula Yaotu Granules for lumbar disc herniation has achieved good clinical results;however, the underlying mechanisms remain poorly understood. OBJECTIVE:To explore mechanisms underlying the effects of the drug serum of Yaotu Granules on NF-κB signaling pathway. METHODS:Eight New Zealand white rabbits were randomly divided into normal saline group, low-, middle-, and high-dose drug groups. Normal saline, 10, 20, and 40 mL of water decoction of Yaotu Granules were administered intragastrical y in the normal saline, low-, middle-, and high-dose drug groups for 7 days, respectively. The drug serum of Yaotu Granules was prepared using serum samples obtained from the common carotid artery at 2 hours after the last administration. Human nucleus pulposus cel s were maintained in DMEM containing 10%(v/v) drug serum for 24 hours. RESULTS AND CONCLUSION:The expression levels of interleukin (IL)-8, tumor necrosis factor (TNF)-α, p-P65, P50, IκB-α(the NF-κB inhibitor), and IκB kinaseβ(IKK-β) mRNA in human nucleus pulposus cel s in three drug serum groups were significantly decreased compared with the normal saline group;in addition, a similar change in the expression levels of p-P65, P50, IκB-α, and IKK-βprotein between groups were detected. Noticeably, the expression levels of IL-8, TNF-α, p-P65, P50, IκB-α, and IKK-βmRNA, as wel as the expression levels of p-P65, P50, IκB-α, and IKK-βprotein were gradual y decreased with the increased drug dose. Al these results suggest that the drug serum of Yaotu Granules can protect nucleus pulposus cel s and delay their degeneration through blocking NF-κB signaling pathway.
2.Effect of Bushen Huoxue Tongluo Capsule on miR-27a in the synovium and chondrocytes in Sprague-Dawley rats with osteoarthritis
Zhitao SUN ; Juyi LAI ; Shenghua HE ; Yeguang WANG ; Jian WANG ; Hualong FENG ; Feiqiang HUANG
Chinese Journal of Tissue Engineering Research 2017;21(16):2484-2488
BACKGROUND: Bushen Huoxue Tongluo Capsule, a well-known traditional prescription, exhibits remarkable treatment outcomes in knee osteoarthritis, but the underlying mechanism is not fully understood. OBJECTIVE: To explore the regulatory effect of Bushen Huoxue Tongluo Capsule on miR-27a of synovial cells in Sprague-Dawley rats with knee osteoarthritis. METHODS: The model of knee osteoarthritis was established in 120 Sprague-Dawley rats using Hulth method, and then these rat models were randomized into four groups, followed by given75 mg/kg normal saline (blank control group), 34 (low-dose group), 75 (medium-dose group) and 140 (high-dose group) mg/kg Bushen Huoxue Tongluo Capsule aqueous solvent via gavage according to body mass, respectively, twice daily, for consecutive 2 courses (15 days as a course) with 2 days in between. The synovium and cartilages were removed after each course, and the expression of miR-27a in the knee joint synovium and chondrocytes was determined by real-time quantitative PCR. RESULTS AND CONCLUSION: Gross observation found that the degeneration of synovium and cartilage in the Bushen Huoxue Tongluo Capsule groups was milder than that in the blank control group. Real-time PCR revealed that there was an increasing trend in the expression of miRNA-27a in the synovium and cartilage in each group, especially in the high-dose group, and there was a significant difference between groups (P < 0.05). Our findings show that Bushen Huoxue Tongluo Capsule can suppress the reduced expression of miRNA-27a in the synovium and chondrocytes in rats with knee osteoarthritis, further alleviating knee degeneration and protecting the knee from damage.
3.Preparation of intervertebral disc degeneration models by percutaneous acupuncture rotary-cut under different anesthesia methods
Juyi LAI ; Shenghua HE ; Zhitao SUN ; Yeguang WANG ; Jian WANG ; Hualong FENG ; Feiqiang HUANG
Chinese Journal of Tissue Engineering Research 2017;21(24):3839-3844
BACKGROUND:Different anesthesia methods significantly impact mean arterial pressure, systolic and diastolic blood pressure, arterial blood pH and blood viscosity, but what kind of anesthesia method is more suitable for preparation of animal model does not reach an agreement. OBJECTIVE: To compare the effectiveness of local anesthesia and general anesthesia for preparation of rabbit intervertebral disc degeneration model in procession of percutaneous acupuncture rotary cut. METHODS:A total of 48 New Zealand white rabbits were randomly divided into the local anesthesia group and the general anesthesia group. The local anesthesia group was anesthetized with 0.5% lidocaine. The general anesthesia group was intraperitoneally injected with 3% sodium pentobarbital (30 mg/kg). In the two groups, models of intervertebral disc degeneration were established by minimally invasive percutaneous acupuncture rotary cut at L4/5, L5/6 levels. Modeling time was observed in both groups. Disc degeneration was evaluated by the gross observation, MRI detection, and histopathological examination at 4, 8, 12, and 16 weeks after modeling. RESULTS AND CONCLUSION: (1) General observation: the nucleus pulposus gradually with darker color and elasticity reduced in two groups after modeling. (2) MRI T2-weighted image results showed no significant changes in disc signal intensity at early stage, but the signal strength showed a decreasing trend with time. (3) Disc degeneration was evaluated according to Pfirrmann grading. Disc degeneration was gradually increased with time in two groups (P< 0.05), but there was no significant difference at postoperative time points in both groups (P > 0.05). (4) Masson staining showed that different degrees of annulus irregular arrangement appear in the two groups 8 and 12 weeks after modeling, but the structure was still intact. 16 weeks later, annulus disorganized, or even breakage in the two groups; no significant difference was detected between the two groups. (5) Safranin O staining showed that nucleus pulposus cells were not significantly reduced in the two groups at 4 weeks, but were significantly reduced at 16 weeks. (6) The average modeling time of local anesthesia group [(15.24±2.67) minutes] was significantly shorter than that in the general anesthesia group [(25.64±6.85) minutes] (P < 0.05). These results indicate that intervertebral disc degeneration model can be successfully established by minimally invasive percutaneous acupuncture rotary cut by using local anesthesia and general anesthesia; however, local anesthesia has a shorter operating time and simple anesthetic procedure. The efficacy was identical between the two methods.
4.Molecular mechanism of Yaotu Granules in the prevention and treatment of lumbar disc degeneration based on RNA-seq technology
Shenghua HE ; Juyi LAI ; Zhitao SUN ; Yeguang WANG ; Jian WANG ; Hualong FENG ; Feiqiang HUANG
Chinese Journal of Tissue Engineering Research 2017;21(24):3778-3783
BACKGROUND:The Herbal Compound formula Yaotu Granules for lumbar disc degeneration has achieved satisfactory efficacy, but the underlying mechanism remains unclear. OBJECTIVE: To explore the possible molecular mechanisms ofYaotu Granules for lumbar disc degeneration based on transcriptome sequencing (RNA-seq). METHODS: Ten New Zealand white rabbits were randomly divided into control and experimental groups (n=5 per group), followed by intragastric injection of 20 mL of normal saline and 20 mL of water decoction of Yaotu Granules, respectively, twice daily for consecutive 1 week. The different drug serums were prepared using serum samples obtained from the common carotid artery at 2 hours after the last injection. Human nucleus pulposus cells were cultured in DMEM containing 10% different drug serums to observe the cellular morphology in each group. The cell viability was detected by trypan blue staining. The passage 3 human nucleus pulposus cells were pretreated for 48 hours, the mRNA level was detected using RNA-seq, and the differentially expressed mRNA was screened by RNA-seq. Subsequently, the differential gene expression, Gene Ontology (GO) enrichment analysis and Pathway enrichment analysis were performed. RESULTS AND CONCLUSION: In the experimental group, there were round or fusiform cells with abundant cytoplasm and clear nuclei that showed smooth surface and complete nuclear membrane; elongated rough endoplasmic reticulum arranged regularly, and less mitochondrion, scattered lysosomes and filaments were visible in the cytoplasm. In the control group, spindle and polygonal cells were found, and the large nuclei with 2 or 3 nucleoli and slightly rough surface were observed; there were mildly dilated endoplasmic reticulum, and few mitochondria with incomplete membrane. The adherent rate, generation time and cell viability of passage 1, 2 and 3 cells in the experimental group were significantly better than those in the control group (P < 0.05). Sequencing results found that there were 464 differentially expressed genes including 143 upregulated, and 321 downregulated genes. GO analysis revealed that the differentiated genes were mainly concentrated on cell regulation and metabolism. Pathway analysis found that mainly differentiated genes focused on the metabolism related pathways. These findings suggest that the differentially expressed gene profile of Yaotu Granules for lumbar disc degeneration is obtained by RNA-seq technology.Yaotu Granules mainly upregulate extracellular matrix metabolism-related genes and downregulate polysaccharide synthesis related genes in the prevention and treatment of lumbar disc degeneration, which provides basis for further research.
5.Zero-P anterior cervical fixation system for multilevel cervical myelopathy
Shenghua HE ; Juyi LAI ; Yeguang WANG ; Zhitao SUN ; Jian WANG ; Hualong FENG ; Feiqiang HUANG
Chinese Journal of Tissue Engineering Research 2017;21(11):1695-1700
BACKGROUND: Zero-P anterior cervical fixation system is an important means for treatment of cervical myelopathy in recent years. Zero-P fixation system can achieve effective decompression of spinal cord and nerve, and restore cervical curvature, with good stability.OBJECTIVE: To investigate the biocompatibility of Zero-P for multilevel cervical myelopathy.METHODS: Totally 62 patients with multilevel cervical disease who were treated by anterior cervical discectomy and fusion with Zero-P were selected, including double segments in 47 cases, three segments in 13 cases and four segments in 2 cases. Clinical efficacy was evaluated by pain visual analogue scale score, Japanese Orthopedic Association score, neck disability index score at postoperative 1, 6, 12, 24 months. Simultaneously, incidence of postoperative complications, cervical intervertebral space height and Cobb angle changes were observed.RESULTS AND CONCLUSION: (1) 54 patients were followed 2 years after treatment by clinic or telephone. Mean operative time was (102.00±32.41) minutes; average blood loss was (62.45±18.36) mL. (2) Two patients affected mild throat discomfort at one day after operation, and the symptoms went to lift six days after. The remaining patients did not experience any other complications. (3) Visual analogue scale score and neck disability index score at 1, 6, 12 and 24 months after operation were all less than preoperatively (P < 0.05); Japanese Orthopedic Association scores were higher than preoperatively (P < 0.05); the Cobb angle was improved significantly and greater than before treatment (P < 0.05).Postoperative cervical intervertebral space height was significantly higher than preoperatively (P < 0.05). Japanese Orthopedic Association results showed that among 54 patients, excellent was found in 34 cases, good in 14 cases,average in 6 cases and poor in 0 cases; the excellent and good rate was 89%. (4) Zero-P has a good clinical efficacy for multilevel cervical myelopathy, relieves symptoms significantly and restores cervical curvature and height, and has a good biocompatibility.
6.Analysis on association between emotional quotient,personality trait and interpersonal trust among students in higher university
Chao ZHANG ; Yeguang WU ; Guimei LI ; Xiaonan WEI ; Chaowei WANG ; Lan ZHENG
Journal of Jilin University(Medicine Edition) 2015;(6):1298-1302
Objective To explore the association between the students’emotional quotient (EQ),personality trait and interpersonal trust, and to clarify the potential function of interpersonal trust of higher students. Methods Interpersonal Trust Scale, Emotional Quotient Table and Eysenck Personality Questionnaire (EPQ) were used to investigate 364 students from Guangxi University and Guangxi Teachers Education University.SPSS 18.0 software was used for one-way ANOVA variance analysis,Pearson product-moment correlation analysis and stepwise Multiple Linear regression analysis.Results The differences in interpersonal trust,EQ and personality trait of EPQ had statistical significance between males and females,and the males’scores were higher than those of females (P <0.05,P < 0.01);the differencs in Lie of EPQ between nations had statistical significance and the scores of Dong, Hui and Tujia ethnic minority were higher than those of other nationalities (P < 0.01 ); the difference in personality trait of EPQ between educational background had statistical significances, and the undergraduates’scores were higher than those of master students (P <0.05).EQ was positively correlated with EPQ-E and EPQ-L (r=0.274,P <0.01;r =0.145,P <0.01),and was negatively correlated with EPQ-N and EPQ-P (r=-0.422,P <0.01;r = -0.202,P <0.01);interpersonal trust was negatively correlated with EQ and EPQ-E (r=-0.273,P <0.01;r=-0.169,P <0.01);special trust was negatively correlated with EQ and EPQ-E (r=-0.252,P <0.01;r = - 0.151,P < 0.01);interpersonal trust and general trust were negatively correlated with EPQ-P (r =0.313,P <0.01;r =0.338,P <0.01),and they were positively correlated with EPQ-L (r=-0.201,P <0.01;r = -0.217,P <0.01).Conclusion The students’EQ,personality trait are related to interpersonal trust of the students in Guangxi universities,and they have important prediction effects. Improving students’ EQ and interpersonal trust plays a positive role, whereas, the possible negative effects accompanying should never be neglected.
7.Experimental study on microvascular architecture of human and rabbit flaps with two methods.
Hongwei ZHANG ; Lin PAN ; Hui WANG ; Xiaochen TIAN ; Zhe CAI ; Yeguang SONG
Chinese Journal of Plastic Surgery 2002;18(2):72-73
OBJECTIVETo study effective method of showing the microvascular architecture of human and rabbit skin flaps.
METHODSAccording to endogenous alkaline phosphatase activity, Gomori-Takamatsu method and NBT/BCIP method were used to observe the microvascular architecture.
RESULTSBoth methods had capability to demonstrate the microvascular architecture of the human skin flap, whereas the examining results of rabbits were negative.
CONCLUSIONSGreat difference of endogeneous alkaline phosphatase activity exists in different animals. Gomori-Takamatsu method and NBT/BCIP method are useful in study of microvascular architecture of human skin flaps.
Alkaline Phosphatase ; metabolism ; Animals ; Humans ; Male ; Microcirculation ; anatomy & histology ; Rabbits ; Surgical Flaps ; blood supply