1.Lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 aggravates cartilage damage in a rat model of osteoarthritis
Erping YANG ; Fei PENG ; Jie LIANG ; Yuanli DU
Chinese Journal of Tissue Engineering Research 2016;20(20):2979-2984
BACKGROUND:Emerging evidence demonstrates that low density lipoprotein receptor-related protein 1 (LRP1)isinvolved in lipid metabolism and regulation of inflammatory reaction.
OBJECTIVE:To explore the effectof lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 on cartilage damage and matrix metaloproteinase 13 in a rat model of osteoarthritis, so as to assess the role of low density lipoprotein receptor-related protein 1 in the pathogenesis of osteoarthritis.
METHODS:Sixty-fourSprague-Dawleyratswere included andramdomlydivided into four groups (n=16 for each): negative control group, no surgery; sham-surgery group, onlythearticular cavity of the knee was exposed; osteoarthritisplus shLRP1 group, rat osteoarthritis models were established by cutting anterior cruciate ligament and removing the medial meniscus partly folowed by an intra-articular injection of lentivirus-mediated siRNA at 2 days after surgery, once a week for 2 consecutive weeks; osteoarthritis group,an intra-articular injection of the negative control lentivirus was performed after surgery. Rats inthe four groups started running on theself-made electric treadmil from 5 days after modeling, 30 minutes per day,totaly 500 meters. Cartilage damage and matrix metaloproteinase 13 expression in cartilage tissues were determined at 2, 4, 6 weeks after surgery.
RESULTS AND CONCLUSION:Gross and pathological observations showed that lentivirus-induced knockdown of lowdensity lipoprotein receptor-related protein 1 aggravatedcartilage damage intherat model of osteoarthritis. At 6 weeks after surgery, Mankin’s scoreand matrix metaloproteinase 13 expression inthecartilage tissues in osteoarthritis plus shLRP1 groupweresignificantly increased compared with other three groups (P< 0.05). These findings indicate that a simulation model of osteoarthritis is developed by cutting anterior cruciate ligament and removing the medial meniscus partly combined with running onthe treadmil. Lentivirus-induced knockdown ofLRP1aggravates cartilage damage in a rat model of osteoarthritis
2.Knockdown of low density lipoprotein receptor-related protein 1 in chondrocytes
Erping YANG ; Fei PENG ; Jie LIANG ; Yuanli DU
Chinese Journal of Tissue Engineering Research 2016;20(15):2171-2177
BACKGROUND:Tumor necrosis factor α, as a pathogenic factor, induces the inflammatory reaction mainlyvia the activation of the nuclear factor kappa B signaling pathway. Low density lipoprotein receptor-related protein 1 (LRP1) is involved in the regulation of the inflammatory reaction induced by cytokines.
OBJECTIVE:To study the effect of knockdown of LRP1 on tumor necrosis factor α-induced inflammatory reaction.
METHODS: Primary cultured rat chondrocytes were transfected with lentivirus-mediated RNA interference to knockdown LRP1 gene. Three days after lentivirus transfection, chondrocytes were pretreated with Bay 11-7082 (10 μmol/L) for 30 minutes prior to the addition of tumor necrosis factor α (30 μg/L) for 30 minutes. Signaling protein and mRNA expressions in chondrocytes were detected by western blot assay and real-time PCR analysis, respectively. Chondrocytes were pretreated with or not Bay 11-7082 (10 μmol/L) 30 minutes prior to the addition of tumor necrosis factor α (30 μg/L) for 12 hours after starvation in DMEM for overnight, and the culture medium was colected for ELISA determination of matrix metaloproteinase 13 level.
RESULTS AND CONCLUSION:Tumor necrosis factor α receptor 1 expression was upregulated in chondrocytes after lentivirus-induced knockdown of LRP1. Increased expression of inducible nitric oxide synthase and activation of the nuclear factor kappa B signaling pathway were found after the addition of tumor necrosis factor α in shLRP1 group. Moreover, increased level of matrix metaloproteinase 13 was determined by ELISA. Taken together, knockdown of LRP1 up-regulates the expression of tumor necrosis factor α-induced inducible nitric oxide synthase and matrix metaloproteinase 13 through the activation of the nuclear factor kappa B signaling pathway.
3.Effects of Early Usage with Wulingjun Powder on the Behavior of Rats with Post-traumatic Stress Disor-der and the Level of IL-1βand IL-6 in Their Hippocampi
Fenglei LI ; Fei DU ; Di WU ; Zhengwu PENG
China Pharmacy 2016;27(25):3478-3480
OBJECTIVE:To discuss the effect of early usage with Wulingjun powder on the behavior of rats with post-traumat-ic stress disorder(PTSD)and the level of interleukin-1β(IL-1β)and IL-6 in their hippocampi. METHODS:50 SD rats were ran-domized into control group(normal saline),model group(normal saline),positive control group(paroxetine,10 mg/kg)and the groups of low and high-dose(0.4,0.8 g/kg)Wulingjun powder. The rats in all groups except for control group,were exposed to single prolonged stress for the establishment of PTSD models. Corresponding drugs were given ig immediately after such establish-ment,qd,for 7 consecutive days. An open field test(the times of the rats’entering the central area and the percentage of the time the rats spent in exploring the central area within 10 min were recorded)was conducted 7 days after giving drugs;and an elevated plus-maze test(the times of the rats’in opening arms and the percentage of the time the rats spent in opening arms within 5 min were recorded)was carried out 8 days thereafter,to observe the behavior of the rats. Then the rats were sacrificed to determine the level of IL-1β and IL-6 in their hippocampi by ELISA. RESULTS:Compared to the normal group,the behavioral index of the rats in model group was significantly lower,and the level of IL-1β and IL-6 in their hippocampi was significantly higher(P<0.05 or P<0.01). Compared to model group,the behavioral index of the rats in positive control group and the group of high-dose Wuling-jun powder was significantly higher,and the level of IL-1β and IL-6 in their hippocampi was significantly lower (P<0.05),but no statistically significant difference was shown in all indexes in the group of low-dose Wulingjun powder (P>0.05). CONCLU-SIONS:High-dose Wulingjun power can improve the anxiety-like behavior of the rats with PTSD,which may be related to the re-duction in the level of IL-1βand IL-6 in the hippocampus.
4.Treatment of Helicobacter pylori-infected gastritis in BALB/c mice by HP1188-IgY
Fei HAN ; Zhibang YANG ; Jianying LI ; Zheng ZHOU ; Fangyi PENG ; Hairong JIANG ; Hongxin DU
Chinese Journal of Pathophysiology 2015;(1):148-153
AIM:To evaluate the effects of treatment with HP 1188-immunoglobulin yolk ( HP1188-IgY) on Helicobacter pylori ( H.pylori)-infected gastritis in BALB/c mice.METHODS:BALB/c mice were used to establish an animal model of H.pylori-infected gastritis, and the mice were divided into 8 groups (10 mice per group).Oral antibiotics were used in group 1, 1 mg HP1188-IgY in group 2, 1 mg HP1188-IgY plus 30%sucralfate in group 3, 5 mg HP1188-IgY in group 4, 5 mg HP1188-IgY plus 30%sucralfate in group 5, PBS in group 6, and 30% sucralfate in group 7 with the treatment once per day for 10 d;and 2.5 mg HP1188-IgY was injected hypodermically twice with a 48-h interval in group 8.Another 10 mice were used as normal control in group 9.The planting of bacteria in the stomach was assayed by bacteri-al culture, rapid urease test, PCR and pathological sectioning .RESULTS:Intragastric administration with 1 mg HP1188-IgY plus 30%sucralfate per day effectively cured the injury of gastric mucosa caused by H.pylori infection, and the effect has no significant difference compared with antibiotics (P>0.05).CONCLUSION:We establish a BALB/c mouse mod-el infected with H.pylori successfully.Sucralfate (30%) is an ideal protectant for HP1188-IgY, which might decrease H. pylori infection in the stomach of BALB/c mice by oral inoculation .
5.Choice and Practice of the new transform moed of science and technology achievemevts of military medicine
Zhongwu LIN ; Yi PENG ; Bin TAN ; Shinan CAO ; Junjie MA ; Fei DU ; Zhongze WANG
Chinese Journal of Medical Science Research Management 2014;27(5):541-543,553
With the development of civil military integration,military scientific research institu tes are facing the challenge of constructing a new mode of translating scientific and technological achievements into practice and enhancing translational efficiency.This paper began with the evolution of translation mode in military institutes and discussed the flaws and insufficiency of current mode,then a triple helix translation mode,which encompass government,industry and research,was introduced and fully explained for future reference.
6.Enhancing chondrogenic differentiation in precartilaginous stem cells with 620 nm red light
Kunpeng LI ; Tao XU ; Yu DU ; Chen GONG ; Fei PENG ; Anmin CHEN ; Fengjin GUO
Chinese Journal of Physical Medicine and Rehabilitation 2012;34(3):172-176
Objective To investigate the effect of 620 nm red light on chondrogenic differentiation in rat precartilaginous stem cells (PSCs). Methods Rats' PSCs were isolated and purified using magnetically activated cell sorting and cultured in vitro.The PSCs were exposed once to 620 nm wavelength red light from a light-emitting diode (LED) with an irradiation energy of 0.5 J/cm2,1 J/cm2,2 J/cm2 or 4 J/cm2.Any effect was confirmed by Alcian blue staining,immunohistochemistry and observing histomorphological changes under a light microscope,as well as detection using a reverse transcription polymerase chain reaction (RT-PCR). Results After being induced for 14 d,the PSCs exhibited polygonal and round shapes. Alcian blue and type Ⅱ collagen immunohistoehemistry staining showed positive results,but the control group had no significant change.RT-PCR showed that the mRNA expression of Sox9 and type Ⅱ collagen increased significantly compared with the control group. Conclusion Low energy 620 nm red light can enhance chondrogenic differentiation in PSCs significantly.
7.A case report of Apert syndrome.
Hong-Hua LI ; Yun-Peng HAO ; Lin DU ; Fei-Yong JIA
Chinese Journal of Contemporary Pediatrics 2011;13(7):604-605
8.Studies on chemical constituents in bark of dictamnus dasycarpus.
Cheng-fang DU ; Xin-xin YANG ; Peng-fei TU
China Journal of Chinese Materia Medica 2005;30(21):1663-1666
OBJECTIVETo study the chemical constituents in bark of Dictamnus dasycarpus.
METHODIsolation and purification were carried out on silica gel column chromatography, prepared thin layer chromatography and sephadex LH - 20, et al. The structures were identified by spectral analysis.
RESULTTwelve compounds were obtained from bark of D. dasycarpus and the structures were determined as dictamnine (I), fraxinellone (II), skimmianine (III), gamma-fagarine (IV ), beta-sitosterol (V), obacunone (VI), limonin disophenol (VII), fraxinellonone (VIII), wogonin (IX), rutevin (X), kihadinin B (XI), dasycarine (XII).
CONCLUSIONCompounds IX and XI were isolated from genus Dictamnus for the first time, and compound VIII was isolated from the species for the first time.
Alkaloids ; chemistry ; isolation & purification ; Dictamnus ; chemistry ; Flavanones ; chemistry ; isolation & purification ; Limonins ; chemistry ; isolation & purification ; Molecular Structure ; Plant Bark ; chemistry ; Plants, Medicinal ; chemistry
9.Paroxysmal Supraventricular Tachycardia in an Infant with Hand, Foot, and Mouth Disease.
Peng HU ; Shu HOU ; Peng Fei DU ; Jia Bin LI ; Ying YE
Annals of Dermatology 2012;24(2):200-202
An 11-month-old male infant was admitted to our hospital with fever, fussiness, poor feeding, vomiting, and tachypnea for two days prior. Physical examination revealed sporadic papules and vesicles occurring on his hands, feet, face, and perianal mucosa. Enterovirus 71 was identified from both throat swab and vesicle fluid using virus isolation techniques. The patient's heart rate fluctuated in a very narrow range from 180~210/beats/min regardless of his physiologic state. An electrocardiogram showed P-waves buried within or occurring just after regular, narrow, QRS complexes. The patient was diagnosed as having hand, foot, and mouth disease in combination with paroxysmal supraventricular tachycardia (PSVT). The child recovered well with symptomatic treatment, including intravenous administration of acyclovir, glucocorticoids, immunoglobulin, adenosine, and sotalol. PSVT was terminated within 36 hours of hospitalization. The skin lesions became crusted on the third day, and then proceeded to heal spontaneously. Here we report on this unusual case and review the associated literature.
Acyclovir
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Adenosine
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Administration, Intravenous
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Animals
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Child
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Electrocardiography
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Enterovirus
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Fever
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Foot
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Foot-and-Mouth Disease
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Glucocorticoids
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Hand
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Heart Rate
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Hospitalization
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Humans
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Immunoglobulins
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Infant
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Male
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Mouth
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Mouth Diseases
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Mucous Membrane
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Pharynx
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Physical Examination
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Skin
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Sotalol
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Tachycardia, Supraventricular
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Tachypnea
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Viruses
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Vomiting
10.Migration and distribution of bone marrow stromal cells in injured spinal cord with different transplantation techniques.
Li FAN ; Fei DU ; Bang-chang CHENG ; Hao PENG ; Shi-qing LIU
Chinese Journal of Traumatology 2008;11(2):94-97
OBJECTIVETo study the regularity of migration and distribution of bone marrow stromal cells (BMSCs) in injured spinal cord with intradural space transplantation.
METHODSForty Wistar rats were randomly assigned into 4 groups. The spinal cord injury model was prepared according to the modified Allen method. BMSCs were labeled by CM-Dil. And 5.0 multiply 10(6) cells were transplanted by different channels including intraventricular injection (Group A),injured spinal cord intrathecally injection (Group B), remote intrathecally injection at the L(3)-L(4) level (Group C), and intravenous injection (Group D). Spinal cord was dissected at 24 hours, 1, 2, 3 and 4 weeks after transplantation. Sections of 4 micromolar were cut on a cryostat and observed under fluorescence microscopy.
RESULTSNo fluorescence was observed 24 hours after transplantation in spinal cord injury parenchyma except Group B. One week later, BMSCs in Groups A and C began to migrate to the injured parenchyma; 2-4 weeks later, BMSCs penetrated into the injured parenchyma except Group D. The number of BMSCs decreased at 3-4 weeks after transplantation. The number of cells in Group B decreased faster than that of Groups A and C.
CONCLUSIONSBMSCs transplanted through intraventricular injection, injured spinal cord intrathecally injection and remote intrathecal injection could migrate to the injured parenchyma of spinal cord effectively. The number of BMSCs migrated into injured spinal cord parenchyma is rare by intravenous injection.
Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; methods ; Cell Movement ; physiology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; pathology ; surgery ; Stromal Cells ; cytology ; transplantation