1.Effect of Tripterygium glycosides on expression of hypoxia inducible factor-1α and endothelin-1 in kidney of diabetic rats.
Wei-Dong CHEN ; Bao-Chao CHANG ; Yan ZHANG ; Ping YANG ; Lei LIU
Journal of Southern Medical University 2015;35(4):499-505
OBJECTIVETo observe the effect of Tripterygium glycosides (TG) on the expression of hypoxia-inducible factor-1α and endothelin-1 in the kidney of diabetic rats and explore the possible mechanism underlying the protective effect of TG against diabetic nephropathy.
METHODSSixty 8-week-old male SD rats were randomly divided into normal control group (n=10) and streptozotocin-induced diabetes mellitus (DM) model group (n=50). The diabetic model rats were then randomly divided into DM group, low-dose (8 mg/kg) and high-dose (16 mg/kg) TG treatment groups, and Irbesartan (50 mg/kg) treatment group. After 8 weeks, the levels of blood glucose (BG), 24-h urine protein (24 h Upro), serum creatinine (Scr) and blood urea nitrogen (BUN) were measured. The pathological changes in the renal tissues were examined by optical microscopy, and the mean glomerular area (MGA) and mean glomerular volume (MGV) were measured with pathological image analysis. Immunohistochemical and Western blotting were used to detect the expression of HIF-1α and ET-1 protein in the renal tissue, and their mRNA expressions were detected using real-time fluorescence quantitative PCR.
RESULTSHIF-1α and ET-1 expression increased in the kidney of diabetic rats. Compared with the diabetic model rats, the rats receiving TG and Irbesartan treatment showed decreased levels of Scr, BUN, 24h Upro, MGA and MGV, improved renal histopathology, and reduced expression of HIF-1α and ET-1 mRNA and protein in the renal tissue. These changes were more obvious in high-dose TG treatment group. Correlation analysis showed that the expression of HIF-1α was positively correlated with that of ET-1, and they were both positively correlated with kidney weight index (KW/BW), 24 h Upro, MGA, and MGV.
CONCLUSIONHIF-1α and ET-1 are overexpressed in the kidney of diabetic rats. TG can improve kidney damage in diabetic rats and delay the development of diabetic nephropathy by inhibiting the HIF-1α and ET-1 expression.
Animals ; Biphenyl Compounds ; pharmacology ; Blood Glucose ; Blood Urea Nitrogen ; Creatinine ; blood ; Diabetes Mellitus, Experimental ; metabolism ; Endothelin-1 ; metabolism ; Glycosides ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tetrazoles ; pharmacology ; Tripterygium ; chemistry
2.Key frames extraction and application in intravascular ultrasound pullback sequences based on manifold learning.
Hai-Qun MAO ; Feng YANG ; Mu-Dan LIN ; Zheng HUANG ; Kai CUI ; Xin-Xin WANG
Journal of Southern Medical University 2015;35(4):492-498
OBJECTIVEWe propose an image-based key frames gating method for intravascular ultrasound (IVUS) sequence based on manifold learning to reduce motion artifacts in IVUS longitudinal cuts.
METHODSWe achieved the gating with Laplacian eigenmaps, a manifold learning technique, to determine the low-dimensional manifold embedded in the high-dimensional image space. A distance function was constructed by the low-dimensional feature vectors to reflect the heart movement. The IVUS images were classified as end-diastolic and non-end-diastolic based on the distance function, and the IVUS images collected in end-diastolic stage constitutes the key frames gating sequences.
RESULTSWe tested the algorithm on 13 in vivo clinical IVUS sequences (images 915±142 frames, coronary segments length 15.24±2.37 mm) to calculate the vessel volume, lumen volume, and the mean plaque burden of the original and gated sequences. Statistical results showed that both the vessel volume and lumen volume measured from the gated sequences were significantly smaller than the original ones, indicating that the gated sequences were more stable; the mean plaque burden was comparable between the original and gated sequences to meet the need in clinical diagnosis and treatment. In the longitudinal views, the gated sequences had less saw tooth shape than the original ones with a similar trend and a good continuity. We also compared our method with an existing gating method.
CONCLUSIONThe proposed algorithm is simple and robust, and the gating sequences can effectively reduce motion artifacts in IVUS longitudinal cuts.
Algorithms ; Angiography ; methods ; Artifacts ; Electrocardiography ; Humans ; Motion ; Reproducibility of Results ; Sensitivity and Specificity
3.Activation of phospholipase C-independent protein kinase C signaling pathway of parathyroid hormone enhances CITED1 expression in mouse osteoblasts.
Song HAO ; Yue MENG ; Wei LI ; Shao-Yu HU ; De-Hong YANG
Journal of Southern Medical University 2015;35(4):486-491
OBJECTIVETo explore the functions of phospholipase C (PLC)-independent protein kinase C signaling pathway (PTH/nonPLC/PKC) of parathyroid hormone (PTH) and its role in bone metabolism.
METHODSOsteoblasts isolated from the calvaria of 2- or 3-day-old C57BL mice, identified by alkaline phosphatase staining and Alizarin red staining, were treated for 4 h with 100 nmol/L [Gly(1), Arg(19)]hPTH(1-28) plus 10 nmol/L RP-cAMP, 10 nmol/L [Gly(1), Arg(19)]hPTH(1-34) plus 10 nmol/L RP-cAMP , 10 nmol/L PTH(1-34), or and 0.1% trifluoroacetic acid (TFA). The total RNA was then isolated for screening differentially expressed genes related to PTH/nonPLC/PKC pathway using Affymetrix mouse 12x135K gene expression profile microarray, and the identified genes were confirmed by real-time quantitative PCR. MC3T3-E1 cells treated with [Gly(1), Arg(19)]hPTH(1-28)+RP-cAMP, [Gly(1), Arg(19)]hPTH(1-34)+RP-cAMP, [Gly(1), Arg(19)]hPTH(1-34)+ RP-cAMP +100 nmol/L Go6983, or 0.1% TFA were also examined for GR(1-28)- or GR(1-34)-mediated gene expression changes using real-time quantitative PCR.
RESULTSAlizarin red staining visualized red mineralized nodules in the osteoblasts at 28 days of culture. According to the genechip results, we selected 56 target genes related to PTH/nonPLC/PKC pathway, among which CITED1 showed higher expressions in [Gly(1), Arg(19)]hPTH(1-34)+ RP-cAMP group than in both the control group and [Gly(1), Arg(19)]hPTH(1-28)+RP-cAMP group (P<0.05), and its expression was the highest in PTH(1-34) group (P<0.05). RT-PCR of MC3T3-E1 cells yielded consist results with those in the primary osteoblasts, and the cells treated with Go6983 (a PKC inhibitor) did not show GR(1-28)- or GR(1-34)-mediated differential expression of CITED1.
CONCLUSIONThe activation of PLC-independent protein kinase C signaling pathway of PTH enhances the expression of CITED1 in mouse osteoblasts to mediate the effect of PTH on bone metabolism, and this pathway is not dependent on the activation of PLC or PKA signaling.
Animals ; Cells, Cultured ; Indoles ; Maleimides ; Mice ; Mice, Inbred C57BL ; Nuclear Proteins ; physiology ; Osteoblasts ; physiology ; Parathyroid Hormone ; physiology ; Protein Kinase C ; physiology ; Signal Transduction ; Skull ; Trans-Activators ; physiology ; Type C Phospholipases
4.Comparison of clinical outcomes of blastocysts derived from non-top quality embryos and cleavage-stage high-quality embryos in frozen-thawed embryo transfer cycles.
Li-Juan XU ; Xin CHEN ; Xiao-Long TIAN ; Yu-Dong LIU ; Nan WANG ; De-Sheng YE ; Ping-Ping GUO ; Shi-Ling CHEN
Journal of Southern Medical University 2015;35(4):481-485
OBJECTIVETo explore the developmental potential of embryos at different developmental days and provide evidence for blastocyst culture of non-top quality cleavage stage embryos in frozen-thawed embryo transfer (FET) cycles.
METHODSThe clinical data of 687 FET cycles were retrospectively analyzed. According to the embryo freezing time, the patients were divided into day 5 (D5) blastocyst group (n=87), day 6 (D6) blastocyst group (n=111) and day 3 cleavage-stage embryo (D3) group (n=489) with hormone replacement cycles or natural cycles for endometrial preparation. The clinical pregnancy rates, miscarriage rates, and implantation rates were compared between the 3 groups.
RESULTSThe clinical pregnancy rate, miscarriage rate and implantation rate per transfer were 58.6%, 9.8%, and 42.9% in D5 group, 32.4%, 19.4%, and 23.3% in D6 group, and 44.9%, 16.4%, and 26.9% in D3 group, respectively. The clinical pregnancy rate and implantation rate were significantly higher in D5 group than in the other two groups (P<0.05).
CONCLUSIONThe D5 blastocysts derived from non-top quality D3 embryos after cryopreservation can have better clinical outcomes than those derived from D3 cleavage-stage embryos and D6 blastocysts, and are therefore a better option than D3 cleavage-stage embryos in FET cycles.
Abortion, Spontaneous ; Blastocyst ; Cleavage Stage, Ovum ; Cryopreservation ; Embryo Implantation ; Embryo Transfer ; Female ; Humans ; Pregnancy ; Pregnancy Rate ; Retrospective Studies
5.Incorporation of TV regularization in deconvolution for partial volume correction in PET imaging.
De-Bin HU ; Li-Jun LU ; Yuan-Yuan GAO ; Hou-Jin ZHANG ; Yan-Jiang HAN ; Cheng-Wei GU ; Jian-Hua MA
Journal of Southern Medical University 2015;35(4):474-480
We propose a method using total variation (TV) regularization in deconvolution for partial volume correction in PET imaging. In the degraded image model, we used TV regularization procedure in Van Cittert (VC) and Richardson-Lucy (RL) deconvolution algorithms. These methods were tested in simulated NCAT images and images of NEMA NU4-2008 IQ phantom and tumor-bearing mouse scanned by Simens Invoen microPET. The simulated experiment and tumor-bearing mouse experiment showed that the algorithms using TV regularization provided superior qualitative and quantitative appearance compared with traditional VC and RL algorithms. When the mean intensity of the tumor increased by (10±1.8)%, the SD increase percentage was decreased from 49.98% to 14.26% and from 42.76% to 4.70%, suggesting the efficiency of the proposed algorithms for reducing PVEs in PET.
Algorithms
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Animals
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Image Processing, Computer-Assisted
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Imaging, Three-Dimensional
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Mice
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Phantoms, Imaging
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Positron-Emission Tomography
6.Role of CD44 in monocyte transmigration across Cryptococcus neoformans-infected blood-brain barrier in vitro.
Li-Ke ZHANG ; Jia-Wen QIU ; Xiao-Lu LIANG ; Bao-Yi HUANG ; Yan LI ; Lei DU ; Min LONG ; Jun LUO ; Sheng-He HUANG ; Hong CAO
Journal of Southern Medical University 2015;35(4):468-473
OBJECTIVETo explore the role of CD44 in monocyte adhesion to human brain microvascular endothelial cells (HBMECs) and monocyte migration across an in vitro model of blood-brain barrier (BBB) infected by Cryptococcus neoformans (Cn).
METHODSAn in vitro blood-brain barrier model was constructed using a transwell chamber covered with a HBMEC monolayer. The wild-type strain of Cn B4500FO2, TYCC645#32 strain with CPS1 gene deletion and PCIP strain with CPS1 complementation were chosen to infect the monolayer HBMECs. THP-1 cells were added to the upper chamber of transwell, and the relative migration rate was determined by counting the number of the cells entering the lower chambers. The inhibitory effects of anti-CD44 monoclonal antibody and the CD44 inhibitor bikunin were examined on THP-1 binding to and migration across HBMECs.
RESULTSCn infection of the HBMECs caused markedly enhanced THP-1 cell adhesion and migration across the monolyers (P<0.01) dependent on Cn concentration and exposure time. Addition of anti-CD44 monoclonal antibody and bikunin significantly lowered THP-1 adhesion and migration rates in the BBB model with Cn-infected HBMECs (P<0.01) with a dose dependence of the antibody (within 0-1 µg) and inhibitor (within 0-20 nmol/L). Both THP-1 adhesion rate and migration rate were lowered in the BBB model infected with CPS1 gene-deleted Cn but increased in the model infected with the complemented strain compared with those in the wild-type strain-infected model.
CONCLUSIONIn the in vitro BBB model, CD44 expressed on HBMECs may play an essential role in monocyte adhesion to and migration across the BBB. The capsular hyaluronic acid may mediate Cn-induced monocyte adhesion and migration.
Blood-Brain Barrier ; immunology ; microbiology ; Brain ; cytology ; microbiology ; Cell Line ; Cryptococcosis ; immunology ; Cryptococcus neoformans ; Endothelial Cells ; microbiology ; Humans ; Hyaluronan Receptors ; metabolism ; Monocytes ; cytology
7.Big data: the progress and applications of antibody repertoire sequencing.
Zai-Bao ZHANG ; Qian GUAN ; Zhen-Hai ZHANG
Journal of Southern Medical University 2015;35(4):463-467
Antibody, the major effector in adaptive immunity, plays key roles in protective and pathogenic immune responses. Integrative analyses of antibody development, differentiation, and maturation promote the research in immune mechanism, vaccine design, and therapies for autoimmune disorders. The development of next generation sequencing technologies has enabled large-scale characterization of functional antibody repertoires. With the advantages of next generation sequencing, antibody and antibody repertoire analysis have been successfully used in identification of HIV-1-broadly neutralizing antibodies, design of rationale structure-based vaccine, and development of immunology. With increasing sequence length and precision, improvement of experimental protocols and bioinformatics analyses, and development of single cell sequencing technology, antibody repertoire sequencing will expedite the research in antibody-related immune response, and thus facilitates vaccine design for infectious diseases, clinical diagnosis and interference of autoimmune diseases. This review introduces the technologies, progresses, applications, and caveats of antibody repertoire sequencing.
Antibodies
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chemistry
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Antibodies, Neutralizing
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Antibody Formation
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HIV-1
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High-Throughput Nucleotide Sequencing
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Humans
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Vaccines
8.Copeptin and ischemia modified albumin in early diagnosis and prognosis of myocardial damage in acute organic phosphorus pesticide poisoning.
Jing LI ; Jianjun ZHANG ; Na LI ; Jia LI ; Juan LIU ; Qian LIU
Journal of Southern Medical University 2015;35(3):459-462
OBJECTIVETo assess the value of combined detection of copeptin and ischemia modified albumin (IMA) in early diagnosis and prognostic evaluation of myocardial damage in patients with acute organic phosphorus pesticide poisoning (AOPP).
METHODSA total of 126 AOPP patients were examined for blood copepin and IMA levels and myocardial injury markers within 1 h after admission.
RESULTSCopeptin and IMA levels significantly increased in patients with AOPP compared with those in the control subjects. Copeptin and IMA levels were significantly higher in severe AOPP cases than in mild to moderate cases (P<0.05). Logistic regression analysis showed that increased copeptin and IMA levels and severe complications of AOPP were associated with an increased risk of cardiovascular events.
CONCLUSIONEarly detection of copeptin and IMA levels has important clinical value in early diagnosis and prognostic evaluation of myocardial damage in patients with AOPP, and their levels are positively correlated with the severity of the condition.
Biomarkers ; blood ; Early Diagnosis ; Glycopeptides ; blood ; Humans ; Organophosphate Poisoning ; diagnosis ; Pesticides ; poisoning ; Prognosis ; Serum Albumin ; Serum Albumin, Human
9.Preliminary experience with endoscopic gastric submucosal tumor resection through the submucosal tunnel using double tunnel and double flex endoscope.
Ying XIONG ; Haiqing HU ; Aimin WANG ; Enqiang LINGHU ; Yuanping LI ; Zhiwei ZHANG ; Yan GENG
Journal of Southern Medical University 2015;35(3):455-458
OBJECTIVETo examine the feasibility and safety of gastric submucosal tunnel dissection of gastric submucosal tumors (SMTs) by double tunnel and double flex endoscope.
METHODSFifty patients with gastric SMTs detected by gastric endoscopy and endoscopic ultrasonography between January, 2012 and August, 2013 were enrolled in this study. Using carbon dioxide throughout the procedure, the mucous in the arc was incised along the margins of the lesion to separate the submucosa and create a tunnel. The exposed SMTs were resected completely and the mucosa was covered by endoscopic forceps followed by clipping of the incision. The complication, clinical outcomes, hospital stays and operation time were evaluated.
RESULTSOf the 50 lesions, 50 were located in the gastric fundus, 17 in the gastric antrum and 5 in the gastric body. The lesions were completely resected in all the patients. The diameter of the resected lesions ranged from 0.5 to 2.5 cm (mean 1.1 ± 0.6 cm), and the operation lasted for 35.3 ± 16.2 min (range 23-76 min). In 5 cases (10%), perforation occurred during the operation and was closed by clipping the incision with endoclips after the lesion resection; these patients were discharged after conservative management. Intraoperative bleeding occurred in 16 cases and was successfully managed through endoscopic methods. No delayed postoperative bleeding or perforation occurred in these patients. None of the 48 patients followed up showed tumor recurrence at one year after the operation, and 2 patients were lost for follow up.
CONCLUSIONEndoscopic submucosal dissection of gastric SMTs is effective and safe using double tunnel and double flex endoscope.
Dissection ; Endoscopes ; Endoscopy ; Endosonography ; Gastric Mucosa ; pathology ; surgery ; Humans ; Neoplasm Recurrence, Local ; Stomach Neoplasms ; surgery
10.Expression of miR-200a in colorectal carcinoma cell lines and its effect on LoVo cells.
Gongfa WU ; Haiyan ZHAO ; Nan HE ; Huixia HAN
Journal of Southern Medical University 2015;35(3):450-454
OBJECTIVETo detect miR-200a expression in human colorectal carcinnoma (CRC) cell lines and explore the role of miR-200a in regulating the biological behavior of CRC cells.
METHODSReal-time quantitative RT-PCR (qRT-PCR) was used to detect miR-200a expression levels in 6 CRC cell lines (HCT116, HT29, LS174T, SW480, SW620 and LoVo). miR-200a mimics were transiently transfected into LoVo, and the changes in cell proliferation, apoptosis, migration, and cell-cell adhesion were assessed using CCK-8 assay, TUNEL assay, transwell migration assay, and homogenous adhesion experiment, respectively.
RESULTSThe expression of miR-200a was down-regulated in the 6 CRC cell lines, among which the highly metastatic LoVo cell line showed the lowest expression and the tumorigenic but non-metastatic CRC cell line HCT116 had the highest expression. Overexpression of miR-200a depressed cell proliferation and migration but promoted cell apoptosis and cell-cell adhesion in LoVo cells.
CONCLUSIONmiR-200a plays a role in regulating the invasiveness and metastasis of CRC, and overexpression of miR-200a causes a significant reduction of cell proliferation and migration and promotes apoptosis and cell-cell adhesion in LoVo cells.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; MicroRNAs ; metabolism ; Transfection