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1.
Chinese Critical Care Medicine ; (12): 152-157, 2023.
Article in Chinese | WPRIM | ID: wpr-991994

ABSTRACT

Objective:To observe the expression of deleted in malignant brain tumor protein 1 (DMBT1) in rat acute respiratory distress syndrome (ARDS) model induced by sepsis and its relationship with ARDS related biomarkers.Methods:Forty-eight healthy male rats were randomly divided into sham operation group (Sham group) and ARDS model group, and the rats in each group were further divided into three subgroups at 6, 12 and 24 hours after operation, with 8 rats in each subgroup. The rats in the Sham group were exposed to the cecum only, and sepsis induced ARDS model was reproduced by cecal ligation and puncture (CLP) in the ARDS model group. The general performance was observed at 6, 12, 24 hours after operation. Abdominal aortic blood of rats was collected, and the levels of DMBT1, surfactant-associated protein D (SP-D), vascular endothelial growth factor (VEGF), interleukins (IL-6, IL-10) in serum were determined by enzyme-linked immunosorbent assay (ELISA). The lung tissues were collected, and the lung wet/dry weight (W/D) ratio was determined. The lung tissue pathological changes were observed under light microscope after hematoxylin-eosin (HE) staining, and the lung tissue injury score was evaluated. The expression of DMBT1 protein in lung tissue was determined by Western blotting. The relationship between the serum DMBT1 and SP-D, VEGF, IL-6, IL-10, lung tissue injury score were analyzed by Pearson correlation analysis.Results:Rats in the ARDS model group showed obvious pathological manifestations after operation. The alveolar structure destruction, inflammatory cell infiltration, and alveolar hemorrhage were observed under microscope. Compared with the Sham group, the lung tissue injury score and the lung W/D ratio at 12 hours after operation in the ARDS model group were significantly increased (lung tissue injury score: 3.35±0.13 vs. 1.16±0.07, lung W/D ratio: 5.36±0.44 vs. 4.38±0.35, both P < 0.05), and pulmonary edema was present, which suggested that the ARDS model caused by CLP was successfully reproduced. The results of ELISA and Western blotting showed that the levels of serum DMBT1, SP-D, VEGF and IL-6 in the ARDS model group increased gradually with time, while the level of IL-10 increased first and then decreased. Compared with the Sham group, the levels of DMBT1 in serum and the expressions of DMBT1 protein in lung tissue in the ARDS model group were significantly increased from 6 hours after operation [serum (ng/L) : 231.96±19.17 vs. 187.44±10.19, lung tissue (DMBT1/β-actin): 2.05±0.19 vs. 0.93±0.25, both P < 0.05], and the levels of SP-D, VEGF, IL-6 and IL-10 in serum were significantly increased from 12 hours after operation [SP-D (ng/L): 73.35±8.05 vs. 43.28±5.77, VEGF (ng/L): 89.85±8.47 vs. 43.19±5.11, IL-6 (ng/L): 36.01±2.48 vs. 17.49±1.77, IL-10 (ng/L): 84.55±8.41 vs. 39.83±5.02, all P < 0.05]. Pearson correlation analysis showed that serum DMBT1 was positively correlated with serum SP-D, VEGF, IL-6, IL-10 and lung injury score at 12 hours and 24 hours in the ARDS model group (12 hours: r values were 0.946, 0.942, 0.931, 0.936, 0.748, respectively; 24 hours: r values were 0.892, 0.945, 0.951, 0.918, 0.973, respectively; all P < 0.05). Conclusion:DMBT1 is a novel early biomarker of ARDS by affecting alveolar epithelial cell, alveolar capillary permeability and inflammatory response.

2.
Chinese Journal of Microsurgery ; (6): 656-663, 2022.
Article in Chinese | WPRIM | ID: wpr-995462

ABSTRACT

Objective:To explore the protective effect and mechanism of improved St. Thomas solution on canine skeletal muscle ischemia-reperfusion injury (IRI).Methods:Between March 2021 and September 2021, in the experimental operating room at the Air Force Hospital of the PLA Eastern Theater Command, 16 Beagles were randomly divided into control group, IRI group, IRI+NS group, and improved St. Thomas group, 4 in each group. The canine skeletal muscle IRI model was established, and the canine vital signs were monitored by pre-perfusion with improved St. Thomas perfusate [potassium chloride (KCl), magnesium sulfate (MgSO 4), and NaHCO 3 (pH adjusted)]. The pathological damage of canine skeletal muscle was explored by hematoxylin eosin (HE) staining, electron microscope detection and tissue wet/dry weight ratio, and blood vessel density. Hypoxia performances were detected by labeling blood vessels and hypoxia-inducible factor-1α (HIF-1α). The IRI model of L6 rat myoblasts was established, and the components of St. Thomas perfusion solution were pre incubated to explore the effect on the inhibition of cell proliferation. And by detecting reduced nicotinamide adenine dinucleotide phosphate (NADPH), F2 isoprostane (F2-isoprostane), interleukin 1β(IL-1β), tumour necrosis factor alpha (TNF-α), myeloperoxide enzyme (MPO), glutathione peroxidase (GSH-Px), etc. to explore its protective mechanism. Statistical software SPSS 23.0 was used for statistical analysis, A P<0.05 was set as statistically significant. Results:In the improved St. Thomas group, the vital signs of the dogs were relatively stable, the amount of maintained dopamine was less, the histopathological structure of the gastrocnemius muscle tended to be intact, the swelling of tissue cells and mitochondria was significantly relieved, and the tissue wet/dry weight ratio was less than that in the IRI group ( P=0.046). Pre-incubated with therapeutic doses of MgSO 4 or NaHCO 3, the proliferation rate of L6 cells was higher than that of IRI group ( P<0.01, P=0.005), NADPH ( P=0.004, P=0.001), F2-isoprostane ( P<0.01, P=0.01), IL-1β ( P=0.02, P=0.015), TNF-α ( P<0.01, P<0.01), MPO ( P<0.01, P<0.01) were all lower than those in the IRI group, except GSH-Px that was higher than what in the IRI group ( P<0.01). Conclusion:Pre-perfusion of the improved St. Thomas solution can stabilise the vital signs of dogs in a short period of time. The solution can improve the state of skeletal muscle cells, improve tissue hypoxia, and reduce the damage of skeletal muscle tissue cells through anti-inflammatory and anti-oxidative stress.

3.
Chinese Journal of Trauma ; (12): 458-461, 2022.
Article in Chinese | WPRIM | ID: wpr-932266

ABSTRACT

Osteofacial compartment syndrome (OCS) is one of the serious complications in traumatic orthopedics. If not treated in time, OCS may result in irreversible damage to nerve and muscle,even amputation or death in serious condition. 5P presents to be the classic clinical diagnosis of OCS, but it is highly subjective and cannot timely and accurately judge the progression of the disease. Intracompartment pressure manometry is the main auxiliary method for the diagnosis of OCS. Although there are many manometry methods, there is still no authoritative pressure threshold as the diagnosis standard. Clinicians often aggressively perform fasciotomy to avoid serious complications, leading to unnecessary fasciotomy. The authors retrospectively reviewed the data of patients with OCS treated at Air Force Hospital of Eastern Theater of PLA from March 2010 to March 2020 and found that some patients with OCS had gradual alleviation of clinical symptoms after appropriate conservative treatments such as brace releasing, limb stabilization and swelling subsidence, with no need of fasciotomy. However, the symptoms of some patients progressively aggravated after the above-mentioned traditional treatments and timely fasciotomy was required. The authors graded the severity of OCS and proposed for the first time the OCS grading criteria according to quantitative clinical results and quantitative indicators such as ratio of mean blood flow velocity of bilateral arteries and pulse wave changes, aiming to take corresponding intervention measures for patients with different OCS classifications, carry out more precise treatment and avoid unnecessary fasciotomy.

4.
Chinese Journal of Trauma ; (12): 1078-1082, 2021.
Article in Chinese | WPRIM | ID: wpr-909979

ABSTRACT

Objective:To explore the efficacy of distracting external fixator for tibiofibular fractures combined with osteofascial compartment syndrome.Methods:A retrospective case-control study was conducted to analyze the clinical data of 62 patients with tibiofibular fractures combined with osteofascial compartment syndrome admitted to Air Force Hospital from Eastern Theater of PLA from March 2009 to March 2019, including 47 males and 15 females, aged 20-78 years[(47.1±13.4)years]. There were 30 patients with tibia shaft fractures, 17 with tibia plateau fractures and 15 with tibia distal fractures. The fractures were classified as type 4A in 18 patients, type 4B in 24 and type 4C in 20 according to AO/OT classification. Distracting external fixation was performed for 30 patients(Group A)and calcaneal tuberosity traction for 32 patients(Group B). Levels of alanine aminotransferase(ALT), urea nitrogen(BUN), creatine kinase(CK)and lactate dehydrogenase(LDH)of the injured limb were compared between the two groups during traction. Additionally, the fasciotomy rate, time of damage control treatment(observation interval from trauma to stage II definitive surgery), time of stage II definitive surgery, internal fixation modalities of stage II definitive surgery, rate of needle tract infection and rate of non-planned secondary surgery were compared between the two groups. The limb function was assessed using Johner-Wruhs scoring system at the last follow-up.Results:All patients were followed up for 12-22 months[(15.1±2.7)months]. Level of CK in Group A was 315.6(140.0, 531.5)U/L, significantly lower than that in Group B[465.5(277.0, 1240.5)U/L]( P<0.05). The two groups revealed no statistical differences in levels of BUN, CK and LDH( P>0.05). The fasciotomy rate in Group A[40%(12/30)]was higher than that in Group B[34%(11/32)], but the difference was statistically insignificant( P>0.05). The time of stage II definitive surgery in Group A was(68.5±17.1)minutes, significantly lower than that in Group B[(89.0±15.1)minutes]( P<0.05). The rate of non-planned secondary surgery in Group A[3%(1/30)]was lower than that in Group B[25%(8/32)]( P<0.05). There were no statistically significant differences in time of damage control treatment, internal fixation modalities of stage II definitive surgery and rate of needle infection between the two groups( P>0.05). According to Johner-Wruhs scoring system, the function in Group A were excellent in 17 patients, good in 5, fair in 2 and poor in 6 at the last follow-up, with the excellent rate of 73%. However, the difference was not statistically significant when compared to Group B: excellent in 13 patients, good in 3, fair in 7 and poor in 9, with the excellent rate of 50%( P>0.05). Conclusion:Compared with calcaneal tuberosity traction, the distracting external fixation of tibiofibular fractures combined with osteofascial compartment syndrome can attenuate soft tissue damage during the traction and shorten the time of stage II definitive surgery by maintaining intraoperative fracture reduction.

5.
Journal of Medical Biomechanics ; (6): E365-E370, 2021.
Article in Chinese | WPRIM | ID: wpr-904409

ABSTRACT

Objective To quantitatively judge the degree of tibial bone healing using the finite element wall thickness analysis method, so as to provide an intuitive diagnostic basis for clinical judgment of tibial union and delayed bone healing. Methods After three-dimensional (3D) modeling for the affected and healthy limb side of 48 patients, the maximum wall thickness (MWT) was calculated, and the ratio (B value) was used as a quantitative index of bone healing. When both BMWT2 and BMWT1 were greater than 0.9, bone healing could be judged. When BMWT2 was between 0.9 and 0.7, bone union was judged to be poor, and there was no significant increase in this value after regular reexamination. When BMWT3 was above 0.9 while both BMWT1 and BMWT2 were smaller than 0.7, it could be judged as internal fixation failure, which should be replaced during the second operation. The clinical diagnosis was revised twice, and the final clinical healing results were observed. Results Clinical diagnosis analysis and finite element wall thickness analysis were carried out in 48 patients during each review period, and 21 cases of delayed bone healing and 27 cases of bone nonunion were judged clinically. Among them, 2 cases were judged to be ineffective, and bone grafting intervention was adopted to replace the internal fixation, 12 cases were judged to be still effective, and all cases were finally healed by surgical intervention of bone grafting alone. By Bowker test, P=0.094 was obtained, indicating that the wall thickness analysis method was consistent with the clinical diagnosis. Conclusions The wall thickness analysis method can be used to quantitatively analyze the degree of bone healing at fracture end and realize the rapid calculation of bone healing degree. The case results in this study show that the finite element wall thickness analysis method is superior to the simple clinical diagnosis method, and has better differential diagnostic significance for early diagnosis of poor bone healing.

6.
Chinese Journal of Digestion ; (12): E002-E002, 2020.
Article in Chinese | WPRIM | ID: wpr-811653

ABSTRACT

The novel coronavirus pneumonia (NCP) has spread from Wuhan to all parts of China since December 2019, and the prevention and control of NCP is a top priority for medical staff. Now report three cases of NCP patients, whose viral nucleic acids still positive in stool after throat swab detection turned negative. In view of the highly homologous and similar clinical manifestations between the 2019 novel coronavirus (2019-nCoV) and the severe acute respiratory syndrome(SARS) related coronaviruses, it is recommended to attach great importance to the detection of the viral nucleic acids in stool, with the reference of SARS prevention and control experience. In order to minimize the risks of gastrointestinal spread, the detection of 2019-nCoV nucleic acids in stool may be recommended as the reference standard of disisolation and discharge.

7.
Chinese Journal of Orthopaedic Trauma ; (12): 746-752, 2020.
Article in Chinese | WPRIM | ID: wpr-867936

ABSTRACT

Objective:To explore the feasibility and effectiveness of four-dimensional analysis of wall thickness for judgment of femoral bone healing.Methods:The clinical data were retrospectively analyzed of the 29 patients who had been diagnosed with femoral bone malunion or bone nonunion at Department of Orthopaedics, Air Force Hospital of Eastern Military Theater of PLA from June 2014 to June 2019. They were 24 males and 5 females with an average age of 41.8 years (from 5 to 54 years). There were 25 cases of delayed union and 4 cases of nonunion (including 2 ones of hypertrophic nonunion and 2 ones of at-rophic nonunion). Mimics software 2.0 and the four-dimensional analysis of wall thickness were used to simulate and analyze the fracture bone healing of the patients at different time points. In comparison with the CT data of the contralateral healthy limb, the fracture bone healing was judged to assist the diagnosis of femoral union, delayed union and nonunion on the basis of the changing trends with time. The original clinical diagnoses were thus revised and the eventual clinical healing outcomes were observed.Results:All the patients were followed up for 1 to 4 years (mean, 14.9 months). Of them, bone union was clinically diagnosed in 13 (44.8%), delayed bone union in 12 (41.4%) and bone nonunion in 4 (13.8%). According to the judgment by four-dimensional analysis of wall thickness, the clinical diagnoses were modified as follows: 9 cases (31.0%) had bone union, 18 cases (62.1%) delayed bone union, and 2 cases (6.9%) bone nonunion. There were statistically significant differences between the 2 methods ( χ2=15.399, P=0.031). Conclusions:The four-dimensional analysis of wall thickness can be used to analyze quantitatively the femoral bone healing, providing a relatively objective basis for clinical diagnosis of bone nonunion and delayed bone union.

8.
Chinese Journal of Comparative Medicine ; (6): 53-59, 2018.
Article in Chinese | WPRIM | ID: wpr-703341

ABSTRACT

Objective To investigate the role and mechanism of PSGL-1 in development of salt-sensitive hypertension in mice. Methods PSGL-1 knockout(PSGL-1 -/-)and wild type(PSGL-1 +/ +)mice were fed a high salt (6% NaCl)or normal salt(0.4% NaCl)diet for three months. Blood pressure was measured under anesthesia via the carotid artery. The status of tissue inflammation and kidney injury was tested by flow cytometry, immunohistochemistry, and western blotting. Results Compared with mice fed a normal salt diet, PSGL-1 +/ +mice fed a high salt diet for three months showed high blood pressure, increased inflammatory cell infiltration in the aorta and skin, and increased inflammatory cytokine expression(interleukin-6, interleukin-1β, and tumor necrosis factor-α)in the kidney, as well as elevated expression of the kidney injury marker, connective tissue growth factor. In contrast, inflammation and kidney injury were not found in PSGL-1 -/-mice fed a high-salt diet. Conclusions In mice,PSGL-1 via inflammation plays a key role in development of hypertension and kidney injury caused by high salt intake.

9.
Chinese Journal of Comparative Medicine ; (6): 46-52, 2018.
Article in Chinese | WPRIM | ID: wpr-703340

ABSTRACT

Objective To explore the role of Sestrin2 in pulmonary alveolar type II epithelial cell injury induced by cigarette smoking and its mechanism of action. Methods The cell injury model was induced by cigarette smoke extract (CSE)in the human pulmonary alveolar type II epithelial A549 cells. The generation of ROS was detected by DCFDA fluorescence probe. The levels of inflammatory factors TNF-α and IL-8 were determined by ELISA, and the expression of Sestrin2 and the peroxiredoxin,Prx-SO2/3H,was detected by Western blot. In addition,all the events were also measured in the A549 cells which were transfected with Sestrin2 siRNA and treated with azithromycin. Results After the CSE treatment,the expression of Sestrin2 in the A549 cells was decreased, the expression of Prx-SO2/3H was increased, the ROS production,secretion of cytokines TNF-α and IL-8 were increased(P < 0.05). These changes were partly reducedby azithromycin, indicating that azithromycin significantly relieved CSE-induced oxidative stress and inflammatory injury.Silencing of Sestrin2 in the A549 cells result ed in an increase of Prx-SO2/3 H expression, ROS production and the secretionof the cytokines TNF-α and IL-8. However, oxidative stress and inflammatory injury were not alleviated with the addition ofazithromycin in the Sestrin2 siRNA silencing A549 cells. Conclusions Sestrin2 plays an protective role in the pulmonaryalveolar type II epithelial cell injury induced by cigarette smoking through negatively regulating the level of intracellularROS via catalyzing the reduction of the hyperoxidized peroxiredoxin Prx-SO2/3 H.

10.
Chinese Journal of Comparative Medicine ; (6): 39-45, 2018.
Article in Chinese | WPRIM | ID: wpr-703339

ABSTRACT

Objective To examine the role of β-tubulin on the interaction between the cholecystokinin B receptor (CCKBR),dopamine D5 receptor(D5R), and water-sodium metabolism. Methods Normotensive and hypertensive renal proximal tubular cells(RPTC)were equally randomized into three separate groups: a gastrin group, fenoldopam group,and gastrin+nocodazole group. Immunofluorescence was used to determine localization of β-tubulin,CCKBR,and D5R. Western blotting was used to detect CCKBR, D5R, and Na-K-ATP expression. Results Gastrin stimulation in normotensive RPTC increased D5R expression(P < 0.05)and decreased Na-K-ATP expression(P < 0.05). These changes were blocked by a tubulin inhibitor(P < 0.05). However, interaction between CCKBR, D5R, and Na-K-ATP expression was not significantly affected in hypertensive RPTC. Immunofluorescence showed that CCKBR and D5R can induce one another,followed by transport to the plasma membrane, which can prevented by a tubulin inhibitor. Further, tubulin is disordered in hypertensive RPTC,which cannot support intracellular CCKBR and D5R transport. Conclusions tubulin plays a key role in the interaction between CCKBR, D5R, and water-sodium metabolism by improving protein transfer from the cytoplasm to cell membrane.

11.
Chinese Journal of Radiation Oncology ; (6): 822-824, 2018.
Article in Chinese | WPRIM | ID: wpr-708271

ABSTRACT

Objective To evaluate the anxiety and explore relevant risk factors in patients receiving radiotherapy and their relatives,aiming to provide evidence for improving the quality of life. Methods Before radiotherapy,the self-rating anxiety scale (SAS) was utilized to evaluate the anxiety of patients and their relatives. The incidence rate of anxiety was analyzed under the influence from different risk factors. Results A total of 646 participants (463 patients and 183 relatives) were included in this study. The average SAS scores of all participants,patients and family relatives were 41.52±10. 08,41.02±19. 37 and 42.79±11. 56, significantly higher than 37.23±12. 58 for the healthy population in China (P= 0. 000. 0.000,0. 000).For patients aged 11-30,31-50 and 51-90 years,the incidence rate of anxiety was 26%(7/ 26),11. 0%(20/ 182) and 19. 1%(47/ 246),respectively (P= 0. 026).The incidence rate of anxiety for patients with and without tumor recurrence was 27% (13/ 48) and 15. 4% (64/ 415) (P= 0. 040).Seventeen of 63 patients (27%) with metastases experienced anxiety,whereas 60 of 396 patients (15. 2%) without metastases suffered from anxiety (P= 0. 020).The incidence rate of anxiety in patients with three or more chronic diseases was 26%(19/ 74),significantly higher compared with 15. 0%(58/ 387) in those with less comorbidities (P= 0. 024). Multivariate binary logistic regression analysis demonstrated that three or more chronic diseases or serious diseases were high risk factors of anxiety (OR= 1. 92,95%CI:1. 03-3. 567). Conclusions Patients who receive radiotherapy and their relatives are prone to anxiety. Young age,tumor recurrence or metastasis and≥ three comorbidities or severe diseases are the high risk factors of anxiety. It is necessary to evaluate the anxiety of patients and their family relatives before radiotherapy and deliver psychological counseling.

12.
Chinese Journal of Comparative Medicine ; (6): 23-30, 2017.
Article in Chinese | WPRIM | ID: wpr-610202

ABSTRACT

Objective To investigate the role and mechanism of platelet in the development of salt-sensitive hypertension.Methods 25 Dahl salt-sensitive rats (Dahl SS) were divided into three groups: low-salt diet (0.12% NaCl, LS), high-salt diet (8%NaCl, HS) and high-salt diet + platelet inhibitor (8%NaCl+busulfan, HS+bus).Blood pressures were measured by tail-cuff method.After six weeks, animals were sacrificed.Platelet p-selectin expression, platelet cytosolic Ca2+ concentration, platelet-leukocyte aggregation (PLA) in peripheral blood, and immune cells infiltrated on aortic walls were assessed by flow cytometry, and serum IL-6 level was tested by ELISA in vivo.Platelets purified from SD rats were treated with normal salt (0.9%NaCl) and high salt (1.3%NaCl), then the cytosolic Ca2+ concentration and p-selectin expression of platelet were detected.Results We found that Dahl SS rats with high-salt diet, relative to low-salt diet, presented with high blood pressure and increased the ratio of platelet p-selectin expression, Ca2+ concentration.IL-6 level and PLA in peripheral blood, and the number of infiltrated immune cells on aortic walls were also significantly elevated in high-salt diet group.The whole events were ameliorated by the platelet inhibitor busulfan.Cytosolic Ca2+ concentration and p-selectin expression were also increased in purified platelets treated with high salt than those treated with low salt (P < 0.05).Conclusions Our findings suggest that high salt induced platelet activation with increased Ca2+ concentration may play an important role in the development of salt-sensitive hypertension via vascular inflammation.However, the detailed mechanisms of platelet activation and development of high blood pressure via inflammation induced by high salt intake remain to be determined.

13.
Chinese Journal of Comparative Medicine ; (6): 24-28,19, 2015.
Article in Chinese | WPRIM | ID: wpr-600816

ABSTRACT

Objective To determine whether dopamine D5 receptor (D5R) regulates the development of dilated cardiomyopathy (DCM) by inhibiting oxidative stress.Methods We developed heart-specific hD5 mutant (α-MHC-hD5F173L) and wild type (α-MHC-hD5WT) transgenic mice.The NOX2 expression and ROS production were tested in the transgenic mice at three month of age.The α-MHC-hD5F173L mice were treated with either NADPH oxidase inhibitor Apocynin (1mmol/kg/day) or phosphate-buffered saline (PBS) as control by intraperitoneal injection for 4 weeks.After then, the indexes of heart function were measured.The hD5WT and hD5F173L were transfected respectively in rat H9C2 cells, in which ROS production and NOX2 expression were detected at basal level.Results The ROS production and NOX2 expression were higher in the heart of α-MHC-hD5F173L than α-MHC-hD5WT mice.Apocynin treatment improved the heart function of α-MHC-hD5F173L mice.NOX2 expression and ROS production were higher in hD5F173L than hD5WT transfected H9C2 cells.Conclusions Dopomine D5 receptor may prevent DCM development by inhibiting oxidative stress.

14.
Chinese Journal of Microbiology and Immunology ; (12): 806-811, 2015.
Article in Chinese | WPRIM | ID: wpr-483983

ABSTRACT

Objective To clone and express the extracellular portion of mouse Fcγreceptor Ⅱ-b ( FcγRⅡb) and to analyze the functions of the expressed proteins in a mouse model of systemic lupus ery-thematosus ( SLE) .Methods The gene fragment encoding FcγRⅡb was amplified by PCR, and then in-serted into the prokaryotic expression vector pET-32a(+) to construct the recombinant expression plasmid pET-FcγRⅡb.The expression plasmid was identified with restriction enzymes and then sent to the Shanghai Bio-Engineering Co.LTD for further sequencing analysis.The transformed Escherichia coli ( E.coli) BL21 ( DE3) strains carrying the recombinant expression plasmid pET-FcγRⅡb were induced by isopropylβ-D-1-thiogalactoside ( IPTG) .The expressed fusion proteins were analyzed by Western blot assay and purified with purification kits.The immune complex ( IC)-binding ability of FcγRⅡb was measured by ELISA.MRL/lpr mice with SLE in both the prevention (12 weeks old, n=40) and the treatment groups (19 weeks old, n=40) were randomly divided into four groups including 60 μl (4.8 μg) treatment group, 120 μl (9.6 μg) treatment group, 180μl (14.4μg) treatment group and PBS treatment group with 10 mouse in each group. The MRL/lpr mice with SLE were injected with the fusion protein through tail vein once a week for four con-secutive weeks.Serum samples were collected from each mouse after one week of observation.The levels of FcγRⅡb in soluble form in mice form both the prevention and treatment groups as well as the levels of anti-double stranded DNA antibodies were detected by ELISA.Results The gene encoding FcγRⅡb was ampli-fied and the recombinant expression plasmid pET-FcγRⅡb was successfully constructed.The recombinant proteins were expressed in the prokaryotic expression system, and then successfully purified.The recombi-nant proteins could bind to IC.Compared with the corresponding PBS control group, the levels of FcγRⅡb in soluble form were increased in mice from both prevention and treatment groups after treating with various concentrations of the recombinant protein (P<0.05).Significant differences in the levels of FcγRⅡb were found among mice of the same age after treating with different concentrations of the recombinant protein ( P<0.05).Compared with the corresponding PBS control group, the levels of anti-double stranded DNA anti-bodies were decreased in mice from both prevention and treatment groups after treating with various concen-trations of the recombinant protein (P<0.05).The levels of anti-double stranded DNA antibodies were grad-ually decreased along with the increasing dosage of protein (P<0.05).Conclusion The extracellular por-tion of murine FcγRⅡb in soluble form was successfully expressed.The recombinant proteins played a cer-tain role in the prevention and treatment of SLE in a mouse model.

15.
Chinese Journal of Tissue Engineering Research ; (53): 2789-2793, 2015.
Article in Chinese | WPRIM | ID: wpr-464291

ABSTRACT

BACKGROUND:Papain-induced rat knee osteoarthritis is a common modeling method, which can obtain a stable osteoarthritis model. OBJECTIVE:To observe the change of ultrastructure of chondrocytes in the early process of papain-induced rat knee osteoarthritis under transmission electron microscope. METHODS:A total of 18 Sprague-Dawley rats were randomly divided into three groups. Two rats were considered as a normal control group, without intervention. The mixture of papain and L-cysteine was injected in right knee joint cavity of 16 rats to induce osteoarthritis models (osteoarthritis model group). Physiological saline was injected in the left side (physiological saline control group). At 1, 2, 4 and 6 weeks after injection, samples were col ected. Transmission electron microscope was used to observe the change of cartilage ultrastructure of the medial femoral condyle joint. RESULTS AND CONCLUSION:For the normal control group and physiological saline control group, their cytoplasm contained abundant rough endoplasmic reticulum and mitochondria. After 1 week of injection, mitochondria vacuoles and light expanded rough endoplasmic reticulum were visible. Two weeks later, lipid droplets appeared, mitochondria degeneration was distinct, vacuolization was serious and its number was reduced, and rough endoplasmic reticulum expansion was obvious. Four weeks later, lipid droplets became increased, and the number of mitochondria decreased significantly. Most of the rough endoplasmic reticula were highly expanded, and part of the rough endoplasmic reticula were dissolved and fractured. Six weeks later, a number of lipid droplets were visible in cytoplasm, most of the mitochondria disappeared, only a smal number of mitochondria existed, and most of the rough endoplasmic reticula were dissolved and fractured. These results confirmed that cartilage ultrastructure changes gradual y in the early process of papain-induced rat knee osteoarthritis under transmission electron microscope.

16.
Chinese Journal of Comparative Medicine ; (6): 53-58, 2015.
Article in Chinese | WPRIM | ID: wpr-463258

ABSTRACT

Objective The goal of this study is to understand the function of FKBP51 in resistant to high fat diet-induced obesity using FKBP51 knockout ( KO) mice and in vitro adipocyte differentiation.Methods Four-week old male FKBP51 KO and wild type ( WT) mice were fed separately with regular or high fat diet for 6 weeks.The body weight and food consumption were recorded weekly, the energy expenditure differences ( O2 consumption, CO2 production, respiratory exchange ratio, and heat production) of each group were monitored using the MM-100 metabolism cages system for 24 hours, then the liver from the above animals were stained with the Oil red-O to detect the lipid accumulation and the expression of metabolic genes.In addition, induction of adipocyte differentiation of immortalized MEF cells from WT and FKBP51 KO mice were used to observe the effect of FKBP51 gene on lipogenesis.Results Compared to WT mice, FKBP51 KO mice has less weight increment, and less lipid accumulation in the liver, but with no difference on food consumption during high-fat diet fed.Moreover, FKBP51 KO mice exhibited more O2 consumption, CO2 production and heated production under both RD and HF diet conditions.The PEPCK, G6Pase and UCP-1 genes up-regulation.In addition, lipid content was reduced in FKBP51 gene deficient MEF cells after adipocyte differentiation.Conclusions The FKBP51 gene plays an important role in high fat diet-induced obesity through the energy metabolism enhancement and lipogenesis inhibition.

17.
Chinese Journal of Comparative Medicine ; (6): 42-46,41, 2014.
Article in Chinese | WPRIM | ID: wpr-599245

ABSTRACT

Objective To measure the organ weights , blood physiological and biochemical parameters , and immune cells of BALB/cA-nu mice .Methods BALB/cA-nu mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .The percentages of CD 3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were detected by FCM in BALB/cA-nu mice at six weeks of age in terms of its T, B lymphocyte cells and NK cell activity .Results At the same age, the weights of the body , liver, and kidney in male mice are significantly higher than females .The HGB、MCH、HCT in male mice are significantly higher than females (P <0.05).The weights of the lung, liver, and kidney, and the parameters of TP、GLOB、CHO、TBIL、UN are significantly lower at five weeks of age comparing with ten weeks (P <0.05).The BALB/cA-nu mice lacks T cells(0.18 ±0.06)%、CD4+T cells(0.26 ±0.08)%、CD8+T cells(0.13 ±0.04)%、The percentage of B cells is CD19+B cells (30.10 ±2.74)% and B220 +B cells(30.55 ±2.77)%.The percentage of NK cells is (1.35 ±0.29)%, and the percentage of granulocytes is (47.90 ±5.48)%.The BALB/cA-nu mice lack T cells and the group of cells .Conclusion The study suggests that the organ weights , blood physiological and biochemical parameters are affected by age and gender in BALB/cA-nu mice that lack T cells immune function .The studied parameters of BALB/cA-nu mice are similar with the same strain in other countries .

18.
Journal of Chinese Physician ; (12): 721-723, 2014.
Article in Chinese | WPRIM | ID: wpr-452862

ABSTRACT

Objective To investigate the association of hepatitis B virus ( HBV) precore ( preC) /C mutations with the pro-gression of liver disease .Methods The HBV genes of 50 chronic hepatitis B , including 30 HBV e-antigen ( HBeAg)-negative and 20 HBeAg-positive patients, were detected with real-time quantification polymerase chain reaction (RT-PCR) and the direct sequencing method.Results The mutations T1762/A176, T1766/A1768, A1896 , and the levels of HBV viral loads significantly correlated with the disease progression .The HBeAg-negative patients had a higher frequency of mutations at T 1762/A1764 , T1766/A1768 , and A 1896 relative to HBeAg-positive patients .Conclusions Patients with advanced liver diseases and with HBeAg-negativity possibly had multi-mutations at T1762/A1764, T1766/A1768, and A1896 in HBV genomes.

19.
Chinese Journal of Comparative Medicine ; (6): 52-55, 2014.
Article in Chinese | WPRIM | ID: wpr-451316

ABSTRACT

Objective To measure the organ weights , blood physiological and biochemical parameters of KK /Upj-Ay/J.Methods KK/Upj-Ay/J mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .Results Parts of the organ weights , blood physiological and biochemical parameters of different ages and sexes were significant differences .The fasted blood glucose of KK/Upj-Ay/J mice reached 7.0mmol/L at 10 weeks of age .Conclusion The results show that the organ weights , blood physiological and biochemical parameters are affected by age and gender of KK /Upj-Ay/J mice.The fasted blood glucose reached the diabetes level at 10 weeks of age .

20.
Chinese Journal of Comparative Medicine ; (6): 56-61, 2014.
Article in Chinese | WPRIM | ID: wpr-451315

ABSTRACT

Objective To measure the organ weights , blood physiological and biochemical parameters , and immune function of Specific Pathogen Free ( SPF) Rag2 knockout ( KO) mice.Methods Rag2 knockout mice were selected at five and ten weeks , and the organ weights , blood physiological and biochemical parameters were observed .The percentages of CD3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in Rag2 KO mice at six week of age in terms of its T , B lymphocyte function and NK cell activity .Results Among the same sex group of NOD/SCID mice, the weights of brain, lung, spleen, liver, heart, kidney, and the levels of TBIL, WBC in 10 weeks of age are higher than 5 weeks.At the same age, the weights of heart, kidney, liver, spleen, and the levels of AST, ALP, A/G, GLU, PLT, PCT, WBC, and LYM%in male mice are higher than females.The Rag2 KO mice lacks T cells(0.36 ±0.15)%、CD4+T cells(0.21 ±0.06)%、CD8+T cells(0.23 ±0.07)%、CD19+B cells(0.28 ±0.04)%、B220+B cells(2.03 ±0.42)%).The percentage of NK cells is(24.13 ±3.62)%, and the percentage of granulocytes is (57.20 ±3.85)%.Conclusion The study suggests that the organ weights , blood physiological and biochemical parameters are affected by age and gender in Rag2 KO mice, which main biological characteristics are similar with C 57BL/6J mice.The Rag2 KO mice show the deficiency of T , B cells function .

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