1.Change of Peripheral-type Benzodiazepine Receptors in Brain Mitochodria and Platelet Membrane in Aging Rat
Yue ZHAO ; Nan YANG ; Chao JI ; Bo SUN ; Pingping ZUO
Chinese Journal of Rehabilitation Theory and Practice 2010;16(4):305-307
ObjectiveTo evaluate the quantitative and qualitative changes of peripheral-type benzodiazepine recepors (PBRs) in brain mitochodria and in platelet membrane in aging rats. MethodsMale Sprague-Dawley rats were divided into 3- and 24-month groups. All animals were sacrificed by decapitation and the brains were immediately removed. Mitochondrial components from dissected cerebral cortex were isolated. The membrane of platelets from venous blood was prepared by the method of hypotonic hemolysis. The specific binding assay of the radioactive PBRs antagonist [3H]PK11195 to membrane was performed. Scatchard analysis was performed to estimate the equilibrium dissociation constant (Kd) and the maximal binding site density (Bmax). ResultsA significant increase in [3H]PK11195 binding activity in the mitochodria from cerebral cortex in 24-month rats was observed compared to that in 3-month rats(P<0-001). Meanwhile, the Scatchard analysis revealed that there was an increase in Bmax, with a significant increase in Kd in 24-month rats. The same change of [3H]PK11195 binding activity was noted for platelet membrane in 24-month rats(P<0-001).ConclusionThe density of PBRs increases in cortex mitochondria in aging rats, but the binding affinity of PBRs decreases which may be attributable to the progressive pathogenesis of aging in rats. [3H] PK11195 binding activity of platelet membrane might reflect the change of PBRs in the brain tissue.
2.Study for cardiac systolic synchronization in patients with ischemic cardiomyopathy
Chao WAN ; Pin SUN ; Yang JI ; Shanglang CAI
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(3):326-329
Objective:To explore value of pulsed wave (PW) and tissue synchronization imaging (TSI) evaluate cardi-ac systolic synchronization in patients with ischemic cardiomyopathy (ICM) .Methods:A total of 50 ICM patients were enrolled as ICM group and 35 healthy volunteers without organic heart disease were regarded as healthy control group ,PW was used to measure mitral diastolic blood flow duration/RR interval (LVFT/RR) to evaluate synchroni-zation of left atrial and left ventricular synchronization ;interventricular mechanical delay (IVMD) was measured to evaluate left and right ventricular synchronization ;TSI software was used to measure mitral annular mean systolic peak velocity (LV-Sm) ,systolic time to peak (Ts) and Ts standard deviation (Ts-SD) of all segments in order to e-valuate left ventricular systolic synchronization .Results:Compared with healthy control group ,there were signifi-cant reductions in LVFT/RR [ (44.74 ± 1.58)% vs .(41.08 ± 4.65)% ] and LV-Sm [ (9.72 ± 0.53) ms vs .(4.09 ± 1.06) ms] ,and significant rise in IVMD [ (15.51 ± 5.52) ms vs .(41.96 ± 4.20) ms] and Ts-SD [ (16.47 ± 4.16) ms vs .(34.13 ± 11.68) ms] in ICM group , P<0.01 all;Ts of anterior wall ,inferior wall et .6 positions were significantly longer ,and its synchronization significantly reduced , P<0. 05 all ,percentages of the most de-layed systolic part of left ventricle were no significant differences between two groups (P> 0.05) .Conclusion:Compared with healthy control group ,left ventricular total systolic function ,left atrial&ventricular ,left&right ventricular synchronization and systolic synchronization within left ventricular significantly reduce in ICM patients .
3.Creation of an animal model for post-operative adhesion prevention.
Chang REN ; Lan ZHU ; Ji-chao SUN
Acta Academiae Medicinae Sinicae 2012;34(2):109-115
OBJECTIVETo establish a rabbit double uterine horn model for assessing the time-course of pelvic adhesions and evaluating the effectiveness of different anti-adhesive materials in reducing adhesions.
METHODSA total of 330 Japanese white rabbits underwent laparotomy, followed by uterine horn incision. Animals were euthanized after 3, 7, 14, 28, and 42 days, respectively.
RESULTSThe surgical procedure was smooth. Rabbit double uterine horn model was applied for the evaluation of pelvic adhesions in a three-dimension fashion. Each of the three means-gauze abrasion, needle holder clamping, and direct uterine incision induced postoperative pelvic adhesions, among which direct uterine incision was the best to mimic conditions after gynecological operations. Under normal circumstances, degradation of filmy fibrinous adhesions by locally released proteases of the fibrinolytic system occurred within 3 days of injury. The regeneration of the mesothelium was completed within 7 days. Collagen reached its peak by day 14. Anti-adhesive materials were supposed to be completely absorbed by day 28, and their effectiveness in preventing pelvic adhesions was confirmed at this time point. Whether their effect can be maintained after the absorption of the anti-adhesive materials was investigated in 42 days.
CONCLUSIONSThe animal model was successfully established. It well mimics the postoperative pelvic adhesions after direct uterine horn injury and thus is a suitable model for studying site-specific adhesions. Observations on the 3rd, 7th, 14th, 28th, and 42nd post-operative days provided a full picture of the adhesion formation process.
Animals ; Biocompatible Materials ; Disease Models, Animal ; Female ; Laparotomy ; Postoperative Complications ; prevention & control ; Rabbits ; Tissue Adhesions ; etiology ; prevention & control ; Uterus ; surgery
4.Cloning, expression and purification of Nocardia brasiliensis proteion P61 with biological activity
Xingzhao JI ; Lu TANG ; Xuexin HOU ; Lina SUN ; Chao WEI ; Shuai XU ; Chenchen SI ; Zhenjun LI
Chinese Journal of Zoonoses 2017;33(3):260-263
We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.
5.Internal fixation with one-hole microplate for the treatment of collateral ligament injuries of the metacarpophalangeal joint of the thumb combined with fracture.
Xi-Xun WANG ; De-Tao SUN ; Xu-Hui CHEN ; Jun LI ; Yan CUI ; Ji-Chao HU ; Zheng-Hua SHU ; Jian HE ; Chao-Qi DING ; Bo CHEN
China Journal of Orthopaedics and Traumatology 2015;28(3):214-218
OBJECTIVETo study clinical effects of one-hole microplate internal fixation for the treatment of collateral ligament injuries of the metacarpophalangeal joint of the thumb combined with fracture.
METHODSTwenty-two patients (16 males, 6 females) with collateral ligament injuries of the metacarpophalangeal joint of the thumb combined fracture were treated with one-hole microplate internal fixation. The age of the patients ranged from 18 to 53 years old with a mean age of 28.5 years old. The duration from injury to surgery ranged from 2 hours to 2 months, and the mean time was 6 days. All the patients had collateral ligament injuries combined with fracture of the metacarpophalangeal joint of the thumb. Thirteen patients had injuries in the right hand and 9 patients had injuries in the left hand. There were 18 cases of closed wound and 4 cases of open wound. Eighteen patients had fresh injuries (< 2 weeks) and 4 had old injuries (> 2 weeks). Sixteen patients had injuries in the ulnar collateral ligament of the thumb combined with fracture, 6 patients had radial collateral ligament injuries of the thumb combined with fracture, 4 cases of which were complicated with injuries of abductor pollicis brevis and the end of the flexor pollicis brevis tender. The size of the avulsed fragment was about 3.0 mm x 4.0 mm to 6.0 mm x 7.0 mm.
RESULTSThe incisions of 22 patients healed by first intention. The follow-up periods ranged from 6 months to 5 years old,with an average of 2.5 years old. The thumb function was evaluated by Saetta and other evaluation criteria, and 20 patients got an excellent result and 2 good.
CONCLUSIONThe application of one-hole microplate internal fixation in treating collateral ligament injuries with fracture of the metacarpophalangeal joint of the thumb is an effective method.
Adolescent ; Adult ; Bone Plates ; Collateral Ligaments ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Metacarpophalangeal Joint ; injuries ; surgery ; Middle Aged ; Thumb ; injuries ; surgery ; Young Adult
6.Progress in the study of Velvet and LaeA proteins and their relation to the development and bioactive compounds in medicinal fungi.
Zhi-chao XU ; Chao SUN ; Jiang XU ; Xin ZHANG ; Hong-mei LUO ; Ai-jia JI ; Yuan-lei HU ; Jing-yuan SONG ; Shi-lin CHEN
Acta Pharmaceutica Sinica 2014;49(11):1520-1527
The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.
Fungal Proteins
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metabolism
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Fungi
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chemistry
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Gene Expression Regulation, Fungal
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Genes, Regulator
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Protein Structure, Tertiary
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Secondary Metabolism
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Structure-Activity Relationship
7.Progress in the study of Velvet and LaeA proteins and their relation to the development and bioactive compounds in medicinal fungi.
Zhichao XU ; Chao SUN ; Jiang XU ; Xin ZHANG ; Hongmei LUO ; Aijia JI ; Yuanlei HU ; Jingyuan SONG ; Shilin CHEN
Acta Pharmaceutica Sinica 2014;49(11):1520-7
The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.
8.Analysis of protease-activated receptor 2 expression and function in cultured human keratinocytes
Zhonglan SU ; Zhigang BI ; Meihua ZHANG ; Chao JI ; Bin CHEN ; Jiping XIA ; Weiling SUN ; Qian GAO ; Hongwei WANG
Chinese Journal of Dermatology 2012;(12):886-890
Objective To assess the expression pattern of protease-activated receptor 2 (PAR2) in human keratinocytes and to characterize its biological functions in the regulation of skin barrier.Methods Primary human keratinocytes and human N/TERT keratinocytes were used as the subject of this study.The expression and distribution of PAR2 in the keratinocytes were analyzed by using immunoflorescence staining and Western blot.Two different PAR2 agonists,trypsin and a PAR2-activating peptide (AP),as well as a PAR2-antagonistic peptide (H2N-FSLLRY-COOH) and a control peptide were used to induce the activation of PAR2 in the keratinocytes.Then,a fluorescence-based calcium mobilization assay was performed to evaluate the biological function of PAR2.Data were statistically analyzed by one-factor analysis of variance.Results Under normal culture conditions,PAR2 was weakly expressed in keratinocytes,and the expression was unaffected by culture medium composition or culture duration.Calcium mobilization was induced by trypsin of 50-250 nmol/L and the PAR2-activating peptide in a dose-and time-dependent pattern.The maximal activation of PAR2 was observed in keratinocytes treated with the PAR2 agonist HAN-SLIGKV-COOH of 75-250 μmol/L.The PAR2-antagonistic peptide (H2N-FSLLRY-COOH) obviously suppressed the increase in calcium mobilization induced by trypsin,while the control peptide PAR-RAP showed no inductive effect on the PAR2 activation based on the absence of calcium mobilization.The substrate-induced calcium release was complete within 250 seconds,and peaked at 50 seconds after the initial trypsin or PAR-AP stimulation.Moreover,the activation of PAR2 was accompanied by an increase in ERK phosphorylation and elicitation of MAPK signaling pathway in keratinocytes.Conclusions Human keratinocytes positively express PAR2,which can be activated by trypsin and PAR2-activating peptides,and the activation of PAR2 may influence the physiological function of keratinocytes by inducing intracellular calcium release.
9.Advantages and disadvantages of the donor site renovation after the wrap-around flap transfer.
Xi-xun WANG ; Jun LI ; Wen-hai SUN ; Bo CHEN ; Ji-chao HU ; Yong WEI ; Zhe TONG ; Zheng-hua SHU ; Yue PAN ; Chao-qi DING
China Journal of Orthopaedics and Traumatology 2010;23(8):604-605
Adolescent
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Adult
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Female
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Humans
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Male
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Middle Aged
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Surgical Flaps
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Thumb
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surgery
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Toes
10.Somatic mutations of mitochondrial DNA in thyroid papillary carcinoma.
Tuan-qi SUN ; Yi WU ; Qing-hai JI ; Jian-chao BIAN ; Zhuo-ying WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2006;41(10):782-785
OBJECTIVETo analyze the somatic mutations in the D-loop of mtDNA and further evaluate the possibility of mitochondrial genetic instability in thyroid papillary carcinoma.
METHODSHypervariable regions ( HVR-I and HVR-II) in the D-loop of mtDNA from the specimen of 35 thyroid papillary cancers and matched lymphocytes were amplified by PCR, and then were sequenced.
RESULTSComparing the sequences of tumors to those of matched lymphocytes and normal thyroid tissues, 5 somatic mutations in 2 patients (5.7%) were found. Two mutations were insertions of C in a poly-cytidine (nt303) microsatellite, and 3 at positions 73, 152 and 194 in HVR-II. In addition, of the 294 genetic variants detected, 292 were previously recorded polymorphisms, whereas 2 were new polymorphisms (nt324:C-->G, nt16092:T-->A).
CONCLUSIONSMutations in the D-loop of mtDNA were found in thyroid papillary cancers, this mutation rate was lower than the reported rate of alteration in tumors of epithelial origin, and further work is required to elucidate the relationship between this mutations and the development of thyroid papillary carcinoma.
Adenocarcinoma, Papillary ; genetics ; Adolescent ; Adult ; Aged ; Base Sequence ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Polymorphism, Single Nucleotide ; Thyroid Neoplasms ; genetics ; Young Adult