1.Expression,purification and biological activity assays of global human C1 q and tumor necrosis factor related protein-2
Hongbo LI ; Xing HU ; Na LI ; Donghai WU
Chinese Pharmacological Bulletin 2014;(7):1023-1025,1026
Aim To prepare soluble global human C1 q and tumor necrosis factor related protein-2 in Escherichia coli. Methods Recombinant expression plasmid was transformed into strain BL21-codonplus (DE3),and the recombinant protein of Trx-gH2 was expressed by IPTG induction and then purified by Ni-NTA affinity and gel filtration chromatography.Results The purified recombinant Trx-gH2 was shown to be active under in vi-vo and in vitro assay conditions.Conclusion Active recombi-nant global hCTRP2 is efficiently prepared from Escherichia coli protein expression system.
2.Expression of myeloperoxidase, soluble intercellular adhesion molecule-1 and pentraxin-3 in the gingival crevicular fluid during maxillary canine distal movement
Huanxi XU ; Hongbo XING ; Fang MIAO ; Ning LI ; Jingyi QIU ; Juan LI
Chinese Journal of Tissue Engineering Research 2016;(2):208-212
BACKGROUND:To dynamicaly monitor the varying levels of inflammatory factors in the gingival crevicular fluid is helpful to assess the early effect of orthodontic tooth movement. Myeloperoxidase, soluble intercelular adhesion molecule-1, pentraxin 3 are proven to be closely related to inflammation, but it is unclear about the levels of these three kinds of inflammatory factors as wel as association of these three kinds of inflammatory factors with orthodontic tooth. OBJECTIVE:To detect the expression levels of myeloperoxidase, soluble intercelular adhesion molecule-1 and pentraxin-3 in the gingival crevicular fluid during maxilary canine distal movement and to assess their correlation with periodontal disease, canine movement distance and orthodontic force. METHODS:Twenty-one orthodontic patients were enroled and assigned into 150 g (n=12) or 100 g (n=9) groups according to orthodontic force. The gingival crevicular fluid samples of orthodontic patients were colected before and at 4, 12, 24 hours, 7, 14 days after maxilary canine distal movement. Levels of myeloperoxidase, soluble intercelular adhesion molecule-1 and pentraxin-3 in the gingival crevicular fluid were measured and analyzed using ELISA assay. RESULTS AND CONCLUSION: During the distal movement of maxilary canine, under orthodontic force, the level of myeloperoxidase was peaked at 4 hours and then decreased, while the expression level of soluble intercelular adhesion molecule-1 was peaked at 12 hours, and then decreased. Both myeloperoxidase and soluble intercelular adhesion molecule-1 levels returned to normal at 7 days under orthodontic force. The expression level of pentraxin-3 was increased significantly under orthodontic force, peaked at 24 hours, and then decreased gradualy to the normal level at 7 days. In addition, the expression levels of myeloperoxidase, soluble intercelular adhesion molecule-1 and pentraxin-3 in the gingival crevicular fluid were significantly higher under 150 g force than under 100 g force. These findings indicate that detecting varying levels of myeloperoxidase, soluble intercelular adhesion molecule-1 and pentraxin-3 in the gingival crevicular fluid is useful to assess the efficiency of orthodontic treatment and prevent adverse reactions.
3.Role of S3I-201 in tubular interstitial lesion in lupus nephritis
Yunxia DU ; Wei ZHANG ; Hongbo LI ; Xiaojuan FENG ; Qinhua WANG ; Lingling XING ; Hui WANG ; Qingjuan LIU
Chinese Journal of Clinical and Experimental Pathology 2016;32(12):1357-1360
Purpose To investigate the role of S3I-201 on tubular interstitial lesion in lupus nephritis.Methods MRt/MpJ mice were designated as the control group.MRL/lpr nice were randomly divided into LN group,S3I-201 group and DMSO group.The serum and 24 h-urine were collected to detect the serum creatinine,blood urea nitrogen and urine protein.Immunohistochemistry was used to detect the expression of FN.Western blotting analysis was used to determine the expression of E-cadherin,α-SMA,MCP-1,ICAM1,STAT3 and p-STAT3.Results Compared with the expression level in control group,the protein level of α-SMA,MCP-1,ICAM1 and FN were increased in renal tissue of MRL/lpr mice,while the expression of E-cadherin was markedly decreased.And the STAT3 was activated in renal tissue of MRL/lpr mice.The administration of S3I-201 could inhibite the activation of STAT3 and ameliorate the expression of E-cadherin,α-SMA,MCP-1,ICAM-1 and FN.Conclusion S3I-201 can relieve the tubular interstitial leison,which maybe concerned with the phosphorylation of STAT3.
4.Coronary artery imaging with 64-slice spiral CT in atrial fibrillation patients:initial experience
Xuhui ZHOU ; Chaogui YAN ; Hongbo XIE ; Xiangmin LI ; Ziping LI ; Quanfei MENG ; Xing CHEN
Chinese Journal of Radiology 2008;42(5):493-497
Objective To discuss the clinical value of coronary artery imaging using 64-slice spiral CT in patient with atrial fibrillation.Methods The images of 31 patients with atrial fibrillation who underwent contrast-enhanced CT coronary angiography were evaluated.The presence of stenosis on each segment of coronary arteries was recorded and their degree of stenosis was measured using the vessel analysis software.Ten patients additionally underwent conventional coronary angiography.The results of conventional coronary angiography were compared with CT coronary angiography of the 10 patients.Results Image reconstruction was based on absolute timing.The image quality of 364 coronary vessel segments on the images from 31 patients was evaluated and defined as excellent,fine,moderate or poor.The image quality was excellent,fine,moderate and poor in 85,41,5,and 8 vessel segments respectively in patient group with heart rate between 47 beat per minent(bpm)and 69 bpm;and in 63,16,13,and 15 vessel segments respectively in patent group with heart rate between 70 bpm and 79 bpm;and in 46,25,23,and 24 vessel segments in patient group with heart rate between 80 bpm and 105 bpm.There was significant difference among the three patient groups(H=22.08,P<0.01).Comparison was carried out between CT angiographic findings and conventional angiographic findings of the 125 segments of the coronary arteries in the 10 patients who underwent conventional coronary angiography.The sensitivity and specificity of CT angiography for diagnosing vessel with significant coronary stenosis(≥50% narrowing)was 85.0%(17/20)and 95.2%(100/105),respectively.Positive predictive value was 77.3%(17/22),and negative predictive value was 97.1%(100/103).Coronary CTA underestimated the lesions of 3 vessel segments and overestimated the lesions of 5 vessel segments.Conclusion Coronary artery imaging with 64-slice row CT had clinical value for patients with atrial fibrillation.
5.Effect of DRD1, DRD3 gene knockout and double gene knockout on body weight in mice
Bao ZHANG ; Bo XING ; Shuguang WEI ; Xiaoni JIA ; Hongbo ZHANG ; Shengbin LI
Chinese Journal of Comparative Medicine 2014;(10):54-57
Objective To study the effect of dopamine receptors on neurological and physiological activities. Methods Dopamine D1 receptor gene (DRD1) knockout mice and dopamine D3 receptor (DRD3) gene knockout mice were introduced, and double gene knockout mice were bred in our lab.Seven SPF male mice in each group were used in this experiment.The food intake, water intake, body weight gain for 24 hours were tested on the age of 30 d, 50 d, and 70 d and were compared with those of wild type mice.Results DRD1 gene and DRD3 gene showed significant effect on the body weight in mice in age of 21 day and 35 day, but at the age of 90 day, the differences became insignificant among the mice of various genetypes.Conclusions Dopamine may effect on the foraging and satiety in newborn mice through regulating the hypothalamic-pituitary-adrenal ( HPA ) axis activity, and finally leads to a reduced body weight gain in newborn mice and puppies during lactation.Furthermore, DRD1 gene and DRD3 gene may influence on body weight of newborn mice through regulating mothers’ lactation, lead to a lower body weight at ablactation, and compensatory increase of body weight after ablactation.Our results provide a substantial foundation for studying the function and interaction of DRD1 and DRD3 genes.
6.Cellular uptake of Tumor-targeted nanoparticles derived from pullulan acetate conjugate
Yajing JIA ; Hongli CHEN ; Hongbo TANG ; Zhimin ZHOU ; Mingming ZHANG ; Xuekun XING ; Huigen FENG
Journal of Medical Postgraduates 2015;(2):127-130
Objective Folate receptors ( FRs) , overexpressed on the surface of a variety of tumor cells , are potential targets for tumor targeting therapy .This study aimed to prepare an FR-mediated drug nanocarrier with folate conjugated pullulan acetate ( FPA) to target chemotherapeutic agents to FR-overexpressed tumor cells and investigate its tumor-suppressing effect in vitro. Methods The cytotoxicity of epirubicin-loaded FPA nanoparticles ( FPA/EPI NP) against HepG2 and Hela cells was evaluated by MTS assay.The HepG2 and Hela cells were divided into five groups to be treated with NPs (NP control), chlorpromazine, chloro-quine, amiloride, and folate, respectively, followed by detection of the fluorescence intensity by flow cytometry . Results FPA/EPI NP was successfully formulated into NPs , with the mean particle size of (268.5 ±12.0) nm, by dialysis with an almost spherical shape . The drug-loading rate and entrapment rate of FPA/EPI NP were (6.45 ±1.04) and (72.45 ±11.50) %, respectively.The survival rates of the HepG2 and Hela cells were both >95%after 24 hours of incubation with FPA NP at 5, 40, 200, 400 and 1000μg/mL and 90.0%after 72 hours.The survival rates of the HepG2 cells treated with 5, 40, 200, 400 and 1000μg/mL FPA/EPI NP for 24 hours were (92.3 ±5.2), (70.4 ±4.6), (50.0 ±4.0), (41.1 ±4.1) and (27.0 ±3.6) %, respectively.Compared with the NP control group, the Hela cells of the chlorpromazine , amiloride and folate groups showed a significantly lower rate of NP uptake (P<0.05), and so did the HepG2 cells pretreated with chlorpromazine or amilo-ride (P<0.05).At 72 hours, the half maximal inhibitory concentrations (IC50) of FPA/EPI NP against HepG2 and Hela cells were 168 and 105μg/mL, respectively . Conclusion Clathrin-mediated endocytosis and macropinocytosis are involved in the internaliza-tion of FPA/EPI NP in HepG2 cells, while clathrin-and FR-mediated endocytosis in that of Hela cells .FPA NP may serve as a new drug carrier for tumor-targeted therapy .
7.Femoral neck fracture in acetabular protrusions secondary to rheumatoid arthritis: a case report and literature review
Hongbo ZHU ; Dan XING ; Yunfei HOU ; Rujun LI ; Bolong KOU ; Jianhao LIN
Chinese Journal of Orthopaedics 2019;39(3):189-192
The present study shows the case of a patient with acetabular protrusions secondary to rheumatoid arthritis progressing to femoral neck fracture.The patient,a 64 years female,had a history of rheumatoid arthtitis for 38 years.The left hip pain and abnormal sound occurred when hip flexion for picking up.She was diagnosed with rheumatoid arthritis secordary to acetabular pelvic retraction and left femoral neck fracture by medical history,physical examination and imaging.Total hip arthroplasty was performed after preoperative examination.Hip dislocation,femoral head removal,acetabular reconstruction;cup fixation,and bone mass assessment are technical challenges during surgery.Based on literature review,this case is belonging to secondary acetabular pelvic retraction,which may be related to acetabular softening caused by rheumatoid arthritis.Whenthe stress from the femoral head exceeds the endurance of the softened acetabulum,the acetabulum protrudes into the pelvis and gradually wraps around the femoral head.Based on the pathological characteristics,itis speculated that the cause of femoral neck fracture is the direct hit of the femoral neck-acetabular rim during hip flexion.In this case,spiral cup prosthesis was used to achieve both the initial stability of the prosthesis and saving bone mass around the acetabulum.The patient was followed up for 3 months with satisfactory position of prosthesis and joint function.
8.Sequence analysis of OTOF gene in a Chinese pedigree with autosomal dominant auditory neuropathy.
Shuai XU ; Guangqian XING ; Xin CAO ; Zhibin CHEN ; Hongbo CHENG ; Huiqin TIAN ; Qinjun WEI ; Xingkuan BU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2007;21(16):735-737
OBJECTIVE:
To investigate if the OTOF gene contributes to the non-syndromic hearing loss of a Chinese pedigree with dominantly inherited auditory neuropathy (AN).
METHOD:
The subjects included were 9 live individuals in an autosomal dominant AN pedigree, 3 sporadic AN patients and 3 normal-hearing controls. Genomic DNA was isolated from the peripheral leukocytes of the subjects using the Pure gene DNA Isolation Kits. Firstly, the whole coding sequence of OTOF gene of one family patient were PCR amplified using specific primers. Each fragment was purified and subsequently analyzed by direct sequencing in an Applied Biosystems 3 730 automated DNA sequencer. The resultant sequence data were compared with the standard sequence to identify deafness-associated mutations. Other DNA samples were then screened for these mutations by PCR amplification and sequence analysis.
RESULT:
PCR amplifications were successfully conducted in all the subjects. Comparison of the resultant OTOF sequence in one family patient with the standard sequence identified 10 nucleotide variants which do not lead to amino acid change. These mutations were also detectable in other family individuals, 3 sporadic AN patients and 3 normal-hearing controls.
CONCLUSION
The OTOF does not seem to contribute to the pathogenesis of this Chinese AN family, which suggest new gene(s) involvement.
Asian Continental Ancestry Group
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genetics
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Base Sequence
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Case-Control Studies
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Chromosome Disorders
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ethnology
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genetics
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DNA Mutational Analysis
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Female
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Genetic Testing
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Hearing Loss
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genetics
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Humans
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Male
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Membrane Proteins
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genetics
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Mutation
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Pedigree
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Sequence Analysis
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Vestibulocochlear Nerve Diseases
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ethnology
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genetics
9.Predictive value of cerebroplacental ratio for perinatal outcomes of induction of labor in prolonged pregnancy
Jianlin ZHAO ; E GONG ; Haijun SHI ; Lan ZHANG ; Xing WANG ; Hongli LIU ; Jie GAN ; Chiying CAO ; Shuai HUANG ; Junnan LI ; Hongbo QI
Chinese Journal of Perinatal Medicine 2021;24(3):209-213
Objective:To investigate the predictive value of cerebroplacental ratio (CPR) for adverse perinatal outcomes of induction of labor in prolonged pregnancy.Methods:This retrospective study recruited 315 singleton pregnant women who had induced labor due to prolonged pregnancy (≥41 gestational weeks) in the First Affiliated Hospital of Chongqing Medical University from January 1, 2019 to April 30, 2020. Based on the occurrence of adverse perinatal outcomes (emergency delivery due to persistent abnormal fetal heart rate monitoring, umbilical artery blood pH at birth <7.2, 5 min Apgar scores<7, transferring to neonatal intensive care unit after birth, chorioamnionitis and vaginal delivery converted to cesarean section), they were divided into two groups: case group ( n=76) and normal group ( n=239). Clinical features and umbilical artery blood flow, middle cerebral artery (MCA) flow and CPR measured in the last ultrasound scan before induction were compared between the two groups using student's t-test, Mann-Whitney U test and Chi-square test. Receiver operating characteristic (ROC) curve was used to analyze the predictive values of umbilical artery blood flow, MCA flow and CPR for the adverse perinatal outcomes. Multivariate logistic regression analysis was used to screen the meaningful predictors. Results:Compared with the normal group, the umbilical artery pulsatility index (PI) (0.9±0.1 vs 0.8±0.1, t=-5.458, P<0.001) and the percentage of abnormal CPR (<1.0) increased significantly [21.1%(16/76) vs 6.3%(15/239), χ2=14.190, P<0.001] in the case group, while the MCA-PI and CPR decreased significantly (1.1±0.2 vs 1.3±0.3, t=5.658, P<0.001; 1.2±0.3 vs 1.6±0.5, t=8.940, P<0.001). The areas under the ROC curves of umbilical artery PI, MCA-PI and CPR for predicting adverse perinatal outcomes were 0.71, 0.71 and 0.77, respectively. CPR had the highest sensitivity (0.74) compared with umbilical artery PI (0.68) and MCA-PI (0.71), but the specificity of them were similar (0.67, 0.66 and 0.66). Multivariate logistic regression analysis showed that only CPR was the independent risk factor for adverse perinatal outcomes ( OR=0.028, 95% CI: 0.010-0.080, P<0.001). Conclusions:As an indicator for early prediction of adverse perinatal outcomes of induction of labor in prolonged pregnancy, CPR was more sensitive but less specific.
10.Investigation of differences between cold and hot nature of Mahuang decoction and Maxing Shigan decoction based on cold/hot plate differentiating assay.
Lei JIA ; Yanling ZHAO ; Xiaoyan XING ; Jiabo WANG ; Wenjun ZOU ; Ruisheng LI ; Hongbo YANG ; Danhong CHENG ; Xiaohe XIAO
China Journal of Chinese Materia Medica 2010;35(20):2741-2744
OBJECTIVETo investigate the objectivity and authenticity of the difference of the Cold and Hot nature between Mahuang decoction and Maxingshigan decoction based on the cold/hot plate differentiating assay.
METHODThe measuring time and dosage of Mahuang decoction and Maxingshigan decoction were investigated before the compliance experiment. Meanwhile, the activities of adenosine triphosphatase were detected in order to probe into the mechanism of their action.
RESULTIt was found that the drug dose should be 10 times amount to human dose (0.80 g x kg(-1) of Mahuang decoction and 2.26 g x kg(-1) of Maxing Shigan decoction) and the measuring time should be from 30 min to 60 min after administering. Compared with the control group, the remaining ratio of the mice treated with Mahuang decoction on the hot pad significantly decreased (P < 0.01). Compared with Mahuang decoction, the remaining ratio of the mice treated with Maxingshigan decoction on the hot pad significantly increased (P < 0.05). Compared with the control group, the Na(+) -K(+) -ATPase activity of liver tissue of the mice treated by Mahuang decoction significantly increased (P < 0.05), while the activity of Na(+) -K(+) -ATPase and Mg(2+) -Ca(2+) -ATPase of liver tissue of the mice treated by Maxingshigan decoction reduced significantly (P < 0.05).
CONCLUSIONThis study showed that the differences of Cold and Hot nature of Mahuang decoction and Maxingshigan decoction were objectively existent from animal ethology, which was consistent with the traditional theroy of Chinese medicine. And the changes of ATPase activity of liver tissue might be the mechanism of drug action.
Animals ; Cold Temperature ; Drugs, Chinese Herbal ; pharmacology ; Ephedra sinica ; Hot Temperature ; Liver ; drug effects ; enzymology ; Male ; Medicine, Chinese Traditional ; Mice ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Thermosensing ; drug effects