1.Expression of a Novo Human Osteoprotegerin-Mycobacterial Heat Shock 70 Fusion Protein with Baculovirus-Insect Expression System
Shu LIU ; Quan-Geng ZHANG ; Yue ZHANG ; Jing MA ; Zhen-Long LIU ; Wen-Ming ZHAO ;
China Biotechnology 2006;0(05):-
Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P
2.Protective effect of rhein lysinate on blood vessel damage induced by oxidative stress in mice and its mechanism
Qiusheng FENG ; Quan KAN ; Cuiping LYU ; Ran LI ; Jingbo WEI ; Yufang ZHAO ; Yongzhan ZHEN
Journal of Jilin University(Medicine Edition) 2015;(6):1171-1175
Objective To investigate the protective effects of rhein lysinate (RHL)on the blood vessel damage induced by oxidative stress in the mice,and to explore its mechanism.Methods The mouse models of oxidative damage were established by intraperitoneal injection of paraquat.30 C57 mice were randomly divided into control, paraquat model,and RHL prevention groups.The mice in RHL prevention group were given RHL by gavage for one week before performing model.The mice in other two groups were given equal volume of distilled water.For making model,paraquat was intraperitoneally injected in the mice in paraquat model and RHL prevention groups once a week for two weeks.The activities of superoxide dismutase (SOD)and glutathione peroxidase (GSH-Px) and the content of serum malonaldehyde (MDA) of the mice were detected 2 weeks after modeling. The pathological profile of blood vessel was observed by hematoxylin and eosin (HE)staining and the level of reactive oxygen species was observed by DCFH-DA staining.The expressions of genes related to blood vessel damage were detected by Western blotting method.Results Compared with control group,the activities of SOD and GSH-Px were decreased and the content of MDA was increased in paraquat model group (P < 0.05 ). Compared with paraquat model group,the activities of SOD and GSH-Px were increased and the content of MDA was decreased in RHL prevention group (P <0.05).The pathological examination indicated the structure of blood vessel of the mice was damaged and the level of reactive oxygen species of blood vessel was increased (P <0.05)in paraquat model group.The pathological changes were significantly improved and the level of reactive oxygen species of blood vessel of the mice was decreased (P < 0.05 )in RHL prevention group. The Western blotting analysis showed that compared with control group,the expression levels of nitric oxide endothelial synthase (eNOS)and caspase-3 of the mice in paraquat model group were decreased (P < 0.05),however the expression level of cleaved fragment of caspase-3 was increased (P < 0.05).Compared with paraquat model group,the expression levels of eNOS and caspase-3 of the mice in RHL prevention group were increased (P < 0.05 )and the expression level of cleaved fragment of caspase-3 was decreased (P <0.05).Conclusion Paraquat could induce vascular cell damage in vivo through increasing the levels of reactive oxygen species, and RHL could antagonize the effects of paraquat by scavenging reactive oxygen species, and up-regulating the eNOS expression and reducing the expression of the cleaved fragment of caspase-3.
3.Comparative observation on efficacy of jieyu pill and maprotiline in treating depression.
Zhen-ming SHEN ; Mei-lan ZHU ; An-quan ZHAO
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(5):415-417
OBJECTIVETo investigate the clinical efficacy, adverse reaction and safety of Jieyu pill (JYP) in treating depression.
METHODSThe randomized controlled trial was conducted in 28 patients in the treated group and 29 patients in the control group treated with maprotiline (Map). The efficacy of treatment was evaluated before treatment and 14, 28 and 42 days after treatment, with Hamilton depression rating scale (HAMD), self-rating scale for depression (SDS), self-rating scale for anxiety (SAS) and clinical global impression (CGI), the adverse reaction was assessed by Asberg Rating Scale (ARS).
RESULTSJYP was effective in treating depression, the markedly effective rate being 78.8%, corresponded to that of Map (82.8%, P > 0.05). After treatment, the scores assessed by HAMD, SDS and SAS were all lower than those before treatment (P < 0.01) respectively, but comparison between the two groups showed insignificant difference (P > 0.05). However, scores of ARS were significantly lower in the treated group than that in the control group, and the efficacy index of JYP was significantly higher than that of Map (P < 0.01).
CONCLUSIONJYP in treating depression shows the efficacy corresponded to that of Map and with less adverse reaction.
Adolescent ; Adult ; Aged ; Antidepressive Agents ; therapeutic use ; Antidepressive Agents, Second-Generation ; therapeutic use ; Depressive Disorder ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Maprotiline ; therapeutic use ; Middle Aged ; Phytotherapy ; Psychiatric Status Rating Scales
4. Detail repair technique for alar deformity after the first stage reconstruction of alar defect
Qinqin ZHAO ; Xiaobo YOU ; Zhen CAI ; Quan LIU
Chinese Journal of Plastic Surgery 2018;34(11):935-938
Objective:
To sum up the repair techniques of secondary alar deformity after the first phase reconstruction of alar defect.
Methods:
From January 2010 to December 2017, 17 patients with secondary alar deformity were included. Secondary alar deformities after the first stage reconstruction of alar defect included the abnormalities of alar groove line, absence of alar-cheek groove and the notch of alar rim. Z-plasty, V-Y advanced flaps, local skin flap and other technologies were introduced to repair details.
Results:
Infection was found in one case of the reconstruction of the alar-cheek groove and the wound was healed by treatment. There was no necrosis occurred in other flap. The shape of the alar was satisfactory and the anterior nostril was not narrow. Patients were followed up for 1 month to 1 year. The average follow-up time was 5 months. No recurrence of nasal alar tumor was found. The patients were satisfied with the appearance.
Conclusions
The symmetrical and satisfactory nasal shape can be obtained with local flaps such as Z-plasty, V-Y advancement of flaps for secondary alar deformities.
5.Progress in preventive effects of cordyceps sinensis on ischemic brain
BAI XUE ; ZHAO YU-QING ; LI CHAO-FENG ; TANG YI-BO ; LIU ZHEN-QUAN
Chinese Journal of Pharmacology and Toxicology 2017;31(10):985-986
Cerebral ischemia is due to cerebral blood supply disorders caused by ischemia and hypoxia resulting in localized ischemic brain necrosis or brain softening of the disease, leading to irreversible brain damage and subsequent loss of neuronal function is a serious threat to human health One of several diseases. For patients with cerebral ischemia, often the lack of effective and extensive treatment. In addition, cerebral ischemia with morbidity, morbidity and mortality are characterized by high, so by the medical profession at home and abroad attention. As a traditional Chinese medicine, cordyceps sinensis (CS) is a complex of ergot fungus, which is parasitized on the larvae of the bat-moth family. The compound is composed of cordycepin, cordyceps polysaccharide, cordyceps sinensis peptides, ergosterol, mannitol, fatty acids and trace elements such as a variety of ingredients, with a wide range of pharmacological effects. Over the years, domestic and foreign scholars on the pharmacological effects of cordyceps sinensis were more comprehensive study of its prevention and treatment of cerebral ischemia is also deepening, found that cordyceps sinensis on cerebral ischemia with anti-inflammatory, reduce oxidative damage and neuronal ischemia damage, reduce neuronal apoptosis, improve memory cognition, reduce thrombosis, inhibit NO production, enhance mitochondrial energy supply, scavenging free radicals and other prevention and treatment. But no relevant review. In this paper, the domestic and foreign literatures on the prevention and treatment of cerebral ischemia by cordyceps sinensis were summarized, analyzed and summarized in order to provide useful information for the research and further development of cordyceps sinensis.
6.Effect of Chinese drugs for strengthening Pi, harmonizing Wei, and dispersing blood stasis on the expression of gastric mucosal heat shock protein 70 in chronic atrophic gastritis patients.
Jia-he THOU ; Zhi-quan FU ; Jian-ping DENG ; Chun-xia LI ; Zhen QIAO ; Wei-qin ZHU ; Hong-wen ZHAO ; Zhen LI
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(4):406-410
OBJECTIVETo observe the effect of Chinese drugs for strengthening Pi, harmonizing Wei, and dispersing blood stasis (CDSPHWDBS) on the expression of gastric mucosal heat shock protein 70 (HSP70) in chronic atrophic gastritis (CAG) patients.
METHODSA total of 100 CAG patients were assigned to the control group and the treatment group by random digit table, 50 in each group. Patients in the control group took Folic Acid Tablet 10 mg each time, 3 times per day. Those in the treatment group took CDSPHWDBS, 100 mL each time, once per day. The treatment course was 6 months for all. Clinical symptoms and signs, endoscopic and histopathological changes were observed before and after treatment in the two groups. The expression of gastric mucosal HSP70 in CAG patients was determined using SP immunohistochemistry. Data were collected by HPIAS-1 000 pathological graphic analysis system, and its expression semi-quantitatively analyzed.
RESULTSThe total effective rate of clinical Chinese medical symptoms and signs was 88. 0% (44/50 cases) in the treatment group and 56. 0% (28/50 cases) in the control group, with significant difference between the two groups (P <0. 01). The improvement rate of endoscopic manifestations such as congestion and edema, erosion, bile regurgitation, pale gastric mucosa, exposed blood vessels, particles proliferation in the treatment group were superior to those in the control group (P <0. 05). The total effective rate of atrophy was 80. 0% (40/50 cases) in the treatment group and 54. 0% (27/50 cases) in the control group, with significant difference between the two groups (P<0. 01). The effective rate of intestinal metaplasia was 75. 0% (12/16 cases) in the treatment group and 33.3% (5/15 cases) in the control group, with significant difference between the two groups (P < 0. 05). The optical density value of gastric mucosal HSP70 was significantly elevated in the two groups after treatment (both P <0. 05). It was higher in the treatment group than in the control group after treatment with significant difference (P <0. 01).
CONCLUSIONCDSPHWDBS had obvious effect in treatment of CAG and could improve pathological changes of precancerous lesions possibly by promoting the expression of gastric mucosal HSP70 in CAG patients.
Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis, Atrophic ; drug therapy ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Immunohistochemistry ; Medicine, East Asian Traditional
7.Expression of FLT3 internal tandem duplication in pediatric patients with acute myeloid leukemia and its correlation with multidrug resistance.
Jiang-Ning ZHAO ; Zhen-Hua QIAO ; Lian-Rong XU ; Quan-Yi LU ; Xiao-Qing NIU ; Peng ZHANG ; Zhao WANG
Journal of Experimental Hematology 2009;17(1):23-26
This study was aimed to investigate the expression of FLT3 internal tandem duplication (FLT3-ITD) in pediatric patients with acute myeloid leukemia (AML) and to analyse the clinical features of patients with mutations and the relation of FLT3-ITD with multidrug resistance gene 1 (mdr1). RT-PCR was used to determine the expressions of FIT3-ITD and mdr1 gene in bone marrow samples from 81 new diagnosed pediatric patients with AML, the cytogenetics and immunophenotypes of bone marrow cells were routinely examined. The results indicated that the FLT3-ITDs were detected in 8 out of 81 pediatric patients (9.88%) and all mutations detected were hybrid, while less frequently this mutation was detected in adult patients. Although they were irrelevant with sex and immunophenotypes, the mutations seemed predominant in older pediatric patients. The leukocyte counts and bone marrow blast cell counts in pediatric patients with FLT3-ITD at diagnosis were higher than those in pediatric patients without FLT3-ITD (p = 0.001 and p = 0.041 respectively), but the normal chromosomes were found in most pediatric patients with FLT-ITD. The patients with FLT3-ITD had lower induction remission rate (only 25%), but the patients without FLT3-ITD had higher remission rate (76.1%). According results detected by RT-PCR, the mdr1 gene was found in 27 pediatric patients, but only 3 out of 8 pediatric patients with FLT3-ITD were detected to express both FLT3-ITD and mdr1, which suggests unrelation between FLT3-ITD occurrence and mdr1 expression. It is concluded that the FLT3-ITD is frequent mutation in pediatric patients with AML, the prognosis is worse and the induction remission rate is lower in these patients, but the FLT3-ITD not relates with the mdr1, which suggests that the common MDR modulators may be un effective for therapy of the patients with FLT3-ITD.
ATP Binding Cassette Transporter, Sub-Family B
;
ATP-Binding Cassette, Sub-Family B, Member 1
;
genetics
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Adolescent
;
Child
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Child, Preschool
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Drug Resistance, Multiple
;
genetics
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Drug Resistance, Neoplasm
;
genetics
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Female
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Gene Duplication
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Humans
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Leukemia, Myeloid, Acute
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genetics
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Male
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Mutation
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Prognosis
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Tandem Repeat Sequences
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fms-Like Tyrosine Kinase 3
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genetics
8.Allergy caused by minidose and low concentration Pingyangmycin: a case report.
Bo-quan SHOU ; Zhao MAO ; Shen-lin ZHANG ; Zhen YANG
West China Journal of Stomatology 2009;27(5):572-573
The emergence of allergy caused by Pingyangmycin is rare. A case of allergy caused by minidose and low concentration Pingyangmycin was reported in this article.
Antibiotics, Antineoplastic
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Bleomycin
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analogs & derivatives
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Humans
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Hypersensitivity
9.Revision of infected total hip replacement:a report of 30 cases
Yu-Feng ZHAO ; Ai-Min WANG ; Hong-Zhen SUN ; Quan-Yin DU ; Qing-Shan GUO ; Zi-Ming WANG ; Si-Yu WU ; Ying TANG ; Yu WANG
Chinese Journal of Trauma 2003;0(11):-
Objective To retrospectively study revisions for infected total hip replacements in 30 cases and discuss the bacteriological characters of the infected total hip replacements,difficulties and strategies in the revision.Methods Thirty revisions of infected total hip replacements were reviewed retrospectively.There were 12 males and 18 females,with mean age of 62.5 years(31-86 years)at revi- sion surgery.Infection was presented one month to four years(mean seven months)after THA operation. The diseases for initial operation included femoral neck fractures in 12 cases,femoral head necrosis in 11,hip osteoarthritis in five and rheumatoid arthritis in two.Twelve eases were treated by one-stage revi- sion and 18 by two-stage revision.Results Before the revision operation,the hip infection were diag- nosed by bacterial culture in 18 cases including five with Staphylococcus epidermidis,four with Staphylo- coccus aureus and nine with other bacteria.Bacteria growth appeared on the specimens from 23 hip joints during the revision surgery but not on the specimens from seven hip joints.Of 12 one-stage revisions,10 cases were followed for mean 16 months,which showed infection recurrence in two eases.Of 18 two-stage revisions,13 cases were followed for mean 20 months,which showed one case with infection recurrence. The mean Harris hip score was improved from preoperative 44 to 84 at follow-up.Conclusions 1) The main bacteria in the infected hip are antibiotic resistant Staphylococcus.2)Because the revision op- eration is difficult,careful preparation before revision is important for success.The fresh surgeon should not attempt.3)The revision strategies should vary according to specific status of the cases.The infection recurrence rate is lower when using a two-stage revision strategy.4)Application of antibiotic bone cement can help improve treatment effect and facilitate functional recovery of the joints.5)The scientific rehabil- itation after operation is very important to functional recovery.
10.Application of microsurgical technique in joint replacement
Yu WANG ; Ai-Min WANG ; Hong-Zhen SUN ; Quan-Yin DU ; Qing-Shan GUO ; Zi-Ming WANG ; Yu-Feng ZHAO ; Si-yu WU ; Ying TANG
Chinese Journal of Microsurgery 2000;0(02):-
Objective To report the experience of the application of microsurgery in joint replace- ment.Methods There were 22 cases,10 cases with segmental acetabular defects treated with the pedicle sartorius muscle iliac bone grafting,5 cases with vascular repair following major vascular injury of extremity during operation,6 cases with neural repair following neural injury during operation,1 case with serious injury reconstruction by elbow joint replacement and free flap.Results The operations succeeded in 22 cases without any postoperative infection.The mean follow-up was 40.1 months (3-72 months) in 22 cases,in which the joint function improved and the operative result was satisfactory with no joint pain.Conclusion Microsurgical technique can reconstruct bone and tissue defect effectively in joint replacement.