1.Research progress on biological characteristics of calcium phosphate cement and its application in stomatology
Haochen GAO ; Jianjin WU ; Zhizhong CAO
Journal of Medical Postgraduates 2015;(8):880-884
Being a new non-ceramic hydroxyapatite artificial bone substitute with biological activity , calcium phosphate cement is widely applied in clinical therapy due to its biocompatibility , osteoconductivity , degradability and possibility of being drug carriers . In addition, it also has a good performance in root canal filling , apexification, plerosis of pulp floor perforation and alveolar defects . However , it is necessary to resolve the issues , such as selecting appropriate hardening time and how to enhance compressive strength . This article presents a review of the research progress on the biological properties and clinical application of calcium phosphate cement in stomatology .
2.Isolation, culture and osteogenic differentiation of mesenchymal stem cells from human umbilical cord: Ultrastructure of cell membrane observed using atomic force microscope
Guodong SUN ; Shixian WU ; Zhizhong LI
Chinese Journal of Tissue Engineering Research 2010;14(1):33-37
BACKGROUND: Mesenchymal stem cells (MSCs) are commonly observed under the scanning electron microscope, transmission electron microscope and inverted microscope. However, above-mentioned observation methods have disadvantages on observing ultrastructure of cell membrane and cytoskeleton. OBJECTIVE: To establish a more effective and appropriate method to isolate, culture and identification of human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and to study the ultrastructure of osteogenic differentiation with the atomic force microscope. METHODS: The hUCMSCs were isolated from human umbilical cord by digested with collagenase. After serial subcultivation in vitro, the stem cells were passaged, and osteogenic differentiation was determined by alkaline phosphatase calcium-cobalt staining and alizarin red staining. hUCMSCs immunophenotype was measured by Flow cytometry.The membrane surface ultrastructure of osteogenic differentiation was observed by Atomic Force Microscope before and after induction. RESULTS AND CONCLUSION: The isolated hUCMSCs by digested with collagenase was efficient. After seeded 24 hours, the adherent cell showed spindle shape, polygonal shape and fibroblast-cell-like shape and the size of hUCMSCs was homogeneous. Flow cytometry analysis revealed that CD29, CD44, CD105 were highly expressed on the surface of passages 3 cells, but there was negative for CD34, CD45 and HLA-DR. These cells were high positive for alkaline phosphate staining and also showed significant calcium node using alizarin red staining after 4 weeks culture induction of osteogenic differentiation. Under atomic force microscopy, undifferentiated stem cells demonstrated insignificant microtubule protrution in a parallel distribution following analysis of cytoskeleton before and after differentiation.
3.Change of hippocampal neurons cholinergic receptor in rats with vascular dementia
Yan XIAO ; Changxue WU ; Zhizhong GUAN
Journal of Clinical Neurology 1997;0(06):-
Objective To explore the change of hippocampal neurons cholinergic receptor in rats with vascular dementia(VD).Methods VD rat models were established by employing improved method of Pulsinelli's four-vessel occlusion.1 month later,the abilities of learning and memory of VD models were tested by Morris water maze.The activities of acetylcholinesterase (AChE) in plasma and hippocampus were detected by the improved Ellman's colorimetric methods.The expressions of hippocampal nicotinic acetylcholine receptor (nAChR) subunits ?3,?4 and ?7 proteins and mRNA were detected respectinely by Western Blotting and RT-PCR,respectively.The results were compared with sham operation group.Results Compared to the sham operation group,the escape latency of Morris water maze test and the first time crossing platform in VD group were significantly prolonged,the number of crossing platform was decreased (all P
4.Association of CYP2D6 gene polymorphism and therapeutic efficacy of venlafaxine on depression in southern region of Fujian
Jiaxin ZHANG ; Zhizhong XU ; Binbin CHEN ; Weige WU ; Jindong CHEN
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(8):687-690
Objective To explore the effect of CYP2D6 gene polymorphism on blood concentration,therapeutic efficacy and adverse effects of anti-depression drug venlafaxine.Methods 69 cases of patients with depression were selected randomly from southern region of FuJian,China.The blood concentration of venlafaxine was detected by HPLC,and the CYP2D6 genotype was determined by sequencing with the amplified PCR products from peripheral blood DNA.The therapeutic efficacy and adverse effects of venlafaxine were evaluated by Hamilton depression scale(HAMD) and treatment emergent symptom scale(TESS) respectively.Results Subjects was divided into CC,CT,TT three groups based on rs16947 locus genotyping.The blood concentrations of venlafaxine were (157.35±15.63) ng/ml,(70.17±5.11) ng/ml,(115.72± 10.2) ng/ml respectively and there was no significant difference (F=1.257,P=0.301).The dose-corrected concentrations and normalized concentrations were not significantly different (F=1.683,1.547,P> 0.05).Similarly,the reduction rate of HAMD (CC:(40.6 ± 7.23) %,CT:(51.7±7.09)%,TT:(42.8±14.1)%) and TESS scores (CC:1.3±0.21,CT:1.3±0.36,TT:1.2±0.28) were not significantly different either (P>0.05).In 69 samples in this study,genotyping of rs3892097 locus only found GG type,and no further examination was performed.Genotyping divided rs1065852 locus into CC,CT,TT three groups.The blood concentrations of venlafaxine were (42.87±9.9) ng/ml,(64.25 ± 13.59) ng/ml,(181.56± 14.15) ng/ml respectively,and there was significant difference among the three groups (F=4.893,P=0.016).The dose-corrected concentrations and normalized concentrations were also significantly different (F=3.985,3.648,P<0.05).The reduction rate of HAMD (CC:(42.6±8.23) %,CT:(48.8± 10.8) %,TT:(63.4±9.15) %) was not significantly different (F=2.961,P=0.07).The difference of TESS scores was similar to that of HAMD reduction rate.Conclusion Among the 3 loci studied,only rs1065852 locus within CYP2D6 gene can affect its enzyme activity and then influence the therapeutic efficacy and adverse effects of venlafaxine.
5.Granulocyte-colony stimulating factor for repair of injured arteries in rats
Jun WU ; Shaoliang CHEN ; Zhizhong LIU ; Xiaobo LI ; Xiaoye SU
Chinese Journal of Tissue Engineering Research 2007;11(23):4630-4633
BACKGROUND:It has been reported that treatment with granulocyte-colony stimulating factor (G-CSF) increases the abundance of circulating CD34+ cells in rats. Data from the study, more important, suggested that mobilized by G-CSF may enhance rapid reendothelization and reduce neointimal formation after vascular injury.OBJECTIVE: To evaluate whether BM-derived CD34+ cells could enhance rapid reendothelization and reduce neointimal formation after balloon-injured carotid artery in an intact rat model.DESIGN: Randomized control animal study.SETTING: Nanjing First Hospital Affiliated to Nanjing Medical University.MATERIALS: The experiment was carried out in Nanjing First Hospital from December 2005 to April 2006. A total of 40 male Sprague-Dawley rats, weighing 200-250 g and of SPF grade, were purchased from National Rodent Laboratory Animal Resources, Shanghai Branch. The recombinant human G-CSF was purchased from Qilu Pharmaceutical. The 2F Fogarty arterial embolectomy catheters were purchased from Edwards Lifesciences. Anti-human CD34 and anti-human CD45 were purchased from Multi Sciences.METHODS: SD rats were divided randomly into treated group (n =20) and control group (n =20). Subcutaneous injection of recombinant human G-CSF (100 μg/kg/day) once daily for 8 days for treated group. Control group as treated with subcutaneous injection of saline. Five days after initiation of G-CSF treatment or saline, the rats were anesthetized by intraperitoneal injection with ketamine. The left common carotid artery was exposed through a midline incision of the ventral side of the neck. A 2F Fogarty arterial embolectomy catheter was inserted through the external carotid artery,inflated with 200 μL air, and passed 3 times along the length of the segment, which was defined proximally by the carotid bifurcation and distally by the edge of the omohyoid muscle. After removal of the catheter, the proximal ligature of the external carotid artery was tied off. ① An average of 1 mL venous blood per rat was collected for enumeration of the white blood wells (WBCs) and CD34+ cells before and 5 days after initiating G-CSF or saline treatment. ② Ten rats in each group were killed with overdose ketamine at 14 and 28 days after balloon injury and left common carotid arteries were harvested. The luminal surface of carotid arteries (n =5, each group) was exposed to calculate the reendothelialized area, which was manually traced with software (Image ProPlus). Reendothelialized area = non-stained with Evans blue area/the total area of balloon-injuried. The cross sections of carotid arteries (n =5, each group) were stained with hematoxylin and eosin (HE) and calculated intima-to-media area ratio (I/M) with software (Image ProPlus) to assess the extent of neointimal thickening. ③ To evaluate the extent of reendothelialization of arteries injury, sections were stained with CD31 and vWF by immunohistochemistry analysis.MAIN OUTCOME MEASURES: ① The number of WBCs and CD34+ cells; ② the extent of reendothelialization of arteries injury; ③ the extent of neointimal hyperplasia (I/M); ④ CD31 + and vWF+ endothelial cells.RESULTS: A total of 40 rats were involved in the final analysis. ① The number of WBCs and CD34+ cells: After 5 days of treatment, the number of WBCs in the treated rats increased more than 2.7-fold compared with control group [(27.60±2.45) ×109 L-1, (10.11±1.81) ×109 L-1, P < 0.01], CD34+ cells increased more than 12.2-fold compared with control group (38.31×107 L-1, 3.14×107 L-1, P < 0.01). ② The extent of reendothelialization: At 14 and 28 days after balloon injury,carotid artery of reendothelialization in the treated group were (68.3±8.3)% and (97.6±4.1)%, superior than the control group (33.8±6.3)% and (76.1±5.2)% (P < 0.01). ③ The extent of neointimal hyperplasia: At 14 and 28 days after balloon injury, the neointima-media (I/M) ratios in the treated rats were 0.39±0.11 and 0.45±0.09, less than the control group 0.87±0.15,1.26±0.16 (P < 0.01). A highly significant inhibition of neointimal hyperplasia was observed in the treated group. ④ CD31+ and vWF+ endothelial cells: At 28 days after injury, sections from G-CSF treated group showed almost complete and continuous monolayer of CD31 and vWF positive cells.In contrast, a patchy and interrupted CD31 and vWF positive cells were found lining the lumen of control group.CONCLUSION:Treatment with G-CSF significantly increases the number of CD34+ cells and accelerates the rate of reendothelialization of injured vessels, leading to marked inhibition of neointimal formation after vascular injury in rats.
6.The clinical diagnostic value of SYT-SSX gene detection combined with immunohistochemistry in synovial sarcoma
Qinzhao YUAN ; Yueqin WU ; Gang XU ; Zhizhong LIANG ; Yanfeng XI
Cancer Research and Clinic 2016;28(4):226-230
Objective To analyse the clinicopathological characteristics of synovial sarcoma (SS) from histomorphology,immunohistochemical (IHC) and fluorescence in situ hybridization (FISH),and to investigate the related factors influencing the prognosis of SS patients.Methods 94 cases were collected,including 60 SS cases and 34 in dined to SS cases.The expressions of common antibodies related to the diagnosis of SS Vimentin,CD99,AE1/AE3,EMA,CD34,Calponin,Ki-67 were detected by IHC,and the SYT-SSX fusion gene was detected by FISH.The clinical pathologic factors that affecting the prognosis of patients were analyzed through the statistical method.Results The positive rates of Vimentin,CD99,AE1/AE3,EMA,CD34,Calponin,Ki-67 were 100.00 %,74.47 %,36.17 %,28.72 %,17.02 %,90.43 % and 60.64 % respectively.FISH results showed that 81 patients were tested with SYT gene translocation,including 54 cases who were considered as SS,and 27 cases who were suspected/inclined to SS,and the number of positive cells surpassed 80 %.The gene translocation was not related with the patient' s age,sex,tumor position,histological type,differentiation of the cancer (all P > 0.05).Single factor survival analysis showed that the degree of differentiation,tumor diameter,the recurrence and metastasis had certain influences on patients' survival time (all P < 0.05).Multiple factors regression analysis showed that the degree of differentiation and tumor diameter were the risk factors influencing the prognosis of SS (risk coefficients > 1,P < 0.05).Conclusions IHC combined with FISH SYT-SSX fusion gene detection have the vital significance in the diagnosis of SS.The degree of differentiation and tumor size are the risk factors influencing the prognosis of SS.
7.Analysis of DNA methylation in peripheral blood of patients with coal-burning-borne endemic fluorosis
Chunyan MAO ; Zhizhong GUAN ; Changxue WU ; Yi LI ; Xiaolan QI
Chinese Journal of Endemiology 2021;40(2):93-98
Objective:To explore the DNA methylation patterns and methylation differential genes of patients with coal-burning-borne endemic fluorosis, and to provide a basis for study of the pathogenesis of fluoride-induced body injury.Methods:A case-control study was conducted in Shuicheng County, Liupanshui, Guizhou Province, ten patients with severe fluorosis were selected as the fluorosis group in Douqing Township, where people burning high fluorine coal in open range all year round; and ten people without fluorosis phenotype were selected as the control group in Huaga Township, where firewood was the main fuel. Peripheral blood samples were collected from the two groups of people. Reduced representation bisulfite sequencing (RRBS) technique was used to detect the whole genome DNA methylation pattern ( n = 4) and DNA differentially methylated region (DMR), the DMR differential degree (log 2Ratio) and KEGG pathway enrichment analysis were used to screen the methylation differential genes, and real-time PCR was used to verify the mRNA expression levels of the candidate methylation differential genes( n = 10). Results:The methylation pattern analysis results showed that the methylation levels of all C bases in the genome DNA of the fluorosis group and the control group were (61.53 ± 0.59)% and (62.48 ± 1.53)%, respectively; among them, the methylated levels at CG sites were (63.75 ± 0.65)%, (64.36 ± 1.01)%, at CHG sites were (13.79 ± 0.72)%, (16.69 ± 4.06)%, and at CHH sites were (25.12 ± 1.72)%, (29.77 ± 3.97)%. Compared with the control group, patients in the fluorosis group had 1 000 DMR distributed on different autosomes; and the chromosome 19 was the most with 104 segments. There were 978 DMR-related genes, including 265 hypermethylation genes and 713 hypomethylation genes; KEGG pathway enrichment analysis showed that methylation differential genes were mainly involved in cell metabolism, cancers, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) and other signaling pathways; combined with the differential degree of DMR, the hypermethylated succinate dehydrogenase complex flavoprotein subunit A pseudogene 3 (SDHAP3, log 2Ratio = 3.487) and hypomethylated nuclear factor κB inhibitor kinase regulatory subunit γ (IKBKG, log 2Ratio =-4.436) were selected as the candidate genes. There were statistically significant differences in the mRNA expression levels of SDHAP3 (0.54 ± 0.08, 1.00 ± 0.00) and IKBKG (1.32 ± 0.39, 1.00 ± 0.00) between fluorosis group and control group ( F = 22.94, 15.09, P < 0.01 or < 0.05). Conclusion:There are a large number of methylation differential genes in the genomes of patients with coal-burning-borne endemic fluorosis and controls, the hypermethylated SDHAP3 and hypomethylated IKBKG may be involved in fluoride induced body injury.
8.136 cases of the kidney injury were analyzed retrospectively
Ke PAN ; Zhizhong TANG ; Kaizhong ZHANG ; Aiming WU
Chinese Journal of Primary Medicine and Pharmacy 2005;0(02):-
Objective To improve the diagnosis and treatment of the kidney injury.Methods 136 cases of the kidney injury were analyzed retrospectively.Results 5 cases died,the other cases were cured.69 cases were followed up for 1~3 years,the cystic kidney function was all normal.Conclusion On time,accurate proceed assessment for the kidney injury,controlling strictly the digit advertises for the operation or not operation is a key to treating kidney injury.
9.The prevention and cure that bleeding after transurethral resection of prostate
Ke PAN ; Zhizhong TANG ; Kaizhong ZHANG ; Aiming WU
Chinese Journal of Primary Medicine and Pharmacy 2006;0(07):-
Objective To explore the prevention and cure methods of the urethra prostate electricity cutting(TURP) on the bleeding after the benign prostatic hyperplasia(BPH) treating.Methods 35 cases with BPH of the bleeding patients after TURP were statistically analyzed.Results 26 examples turned for the better after general treatment;6 examples turned for the better after resectoscope wash gore and gave or got an electric shock the hemostasis;3 examples changed to the surgical operation hemostasis cured.Conclusion Accurate disposal inside operation and process behind operation is a key to decrease bleeding after the TURP of the BPH.
10.Percentage of primary open angle glaucoma in retinal vein occlusion patients
Ke, XU ; Lingling, WU ; Zhizhong, MA ; Yuling, LIU ; Fang, QIAN
Chinese Journal of Experimental Ophthalmology 2016;34(6):552-557
Background Researches determined that the pathogenesis of retinal vein occlusion (RVO) is associated with primary open angle glaucoma (POAG) in Caucasian population.However,the relationship between RVO and POAG in Asian population is unclear.Objective This study was to survey the percentage of POAG in RVO patients in China.Methods A prospective cross-sectional observational study based on hospital was performed.Three hundred and seventy five patients with newly diagnosed RVO by fluorescein fundus angiography (FFA) were consecutively enrolled in Peking University Third Hospital from October 2011 to May 2013,and gonioscopy was carried out in all the patients.Glaucoma was diagnosed according to the criteria of the International Society of Geographical and Epidemiological Ophthalmology (ISGEO).RVO was classified into central retinal vein occlusion (CRVO),hemicentral retinal vein occlusion (HRVO) and branch retinal vein occlusion (BRVO) based on fundus photograph and FFA,or classified into arteriovenous crossing RVO (AV-RVO),optic cup RVO (OC-RVO),optic nerve RVO without optical nerve head swelling (NONHS-RVO) and RVO with optical nerve head swelling (ONHS-RVO) based on the sites of venous occlusion.The percentages of POAG in different types of RVO were calculated.This study was approved by Ethic Committee of Peking University Third Hospital,and written informed consent was obtained from each patient before entering study cohort.Results Three hundred and seventeen RVO patients finished screening of glaucoma,with the response rate 84.5%.The percentage of POAG is 8.2% in RVO patients,including 7.1% in the CRVO patients,23.5% in HRVO patients and 6.2% in BRVO patients.The percentage of POAG was significantly higher in OC-RVO patients (27.9%) and NONHS-RVO patients (13.2%) than that in AV-RVO patients (4.7%) and ONHS-RVO patients (1.3%) (all at P<0.05).Conclusions The overall percentage of POAG in RVO patients in China is 3-4 times higher than general population.Higher percentage of POAG is found in OC-RVO patients and NONHS-RVO patients,suggesting that pathogenesis of RVO is probably associated with the glaucomatous anatomic changes of optical nerve head.