2.Distribution, isolation and culture of epidermal stem cells of rats in vitro
Wei CHEN ; Tian YANG ; Xiaohua LIAN ; Ke YANG ; Enyi HUANG
Journal of Third Military Medical University 1988;0(05):-
Objective To investigate the distribution, isolation and culture of the epidermal stem cells from rats. Methods Immnohistochemical methods were used to confirm the location of the epidermal stem cells. The skin of neonatal rats were dissociated into single cells by dispaseⅡ and trypsin solution,the rapidly adherent cells to collgenⅣ were cultured with KSFM,and those no rapidly adherent cells were regarded as control. Immunohistology and flow cytometry were conducted to identify the epidermal stem cells. Results ? 6-integrin and K15 were expressed in the basal layer cells and hair follicle bulge cells, while the CD71 was negative negatively expressed. CD34 were expressed in the hair follicle bulge cells while not in the basal layer cells. The epidermal stem cells isolated by collgenⅣ had higher colony forming efficiency. Immunocytochemical staining showed that ? 6-integrin and K15 were strongly expressed in the cultured epidermal stem cells. Flow cytometry indicated that 84% cultured epidermal stem cells were expressed ? 6-integrin. Conclusion The epidermal stem cells of rats are located at the basal layer of epidermis and the hair follicle bulge.
3.Feasibility of diagnosing and staging liver fibrosis with diffusion weighted imaging.
Nai-Yi ZHU ; Ke-Min CHEN ; Wei-Min CHAI ; Wei-Xia LI ; Lian-Jun DU
Chinese Medical Sciences Journal 2008;23(3):183-186
OBJECTIVETo assess the clinical feasibility of diagnosing and staging liver fibrosis by apparent diffusion coefficient (ADC).
METHODSTotally, 43 patients (mean age 29.3 years) with chronic hepatitis by liver biopsy and 7 healthy controls (mean age 39.9 years) underwent liver diffusion weighted imaging (DWI) with four b values: 0, 200, 500, and 1000 s/mm2 respectively. The liver fibrosis was staged according to Ishak fibrosis stage. The ADC value of liver fibrosis patients and healthy controls was compared. The correlation of ADC value and liver fibrosis staging was analyzed.
RESULTThe histological staging showed 8 stage 1 patients, 10 stage 2 patients, 6 stage 3 patients, 9 stage 4 patients, 8 stage 5 patients and 2 stage 6 patients. The mean ADC value of liver fibrosis patients was significantly lower than that of healthy controls except for stage 1 group (P < 0.05). There was a negative correlation between liver fibrosis staging and ADC value (r = -0.697 with b=500 s/mm2, P < 0.01). Receiver operating characteristic (ROC) curve of ADC value of advanced liver fibrosis (Ishak stage F3 and higher) showed that area under curve = 0.913, 0.825, and 0.794 with b = 500, 1000, and 200 s/mm2, respectively (95% confidence interval: 83.6%-99.0%, 70.7%-94.3%, 66.5%-92.4%; P < 0.05). When b value was 500 s/mm2, the sensitivity (84%) and specificity (80%) of DWI for diagnosis of advanced liver fibrosis were the highest.
CONCLUSIONDWI is proved to be a useful clinical tool in the quantitative evaluation of liver fibrosis and in the prediction of the process of liver fibrosis with the recommendable b value (500 s/mm2).
Adult ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; pathology ; Male ; Middle Aged ; ROC Curve ; Young Adult
4.Time course of Q value after myopic laser-assisted in situ keratomileusis.
Zheng-Wei ZHANG ; Wei-ran NIU ; Ming-ming MA ; Ke-li-mu JIANG ; Bi-lian KE
Chinese Medical Sciences Journal 2011;26(3):141-145
OBJECTIVETo assess the time course of Q value after myopic laser-assisted in situ keratomileusis (LASIK) and preliminarily evaluate the determinants of the difference of Q value between before and after LASIK.
METHODSWe performed a retrospective, longitudinal investigation on patients undergoing wavefront optimized LASIK therapy for emmetropization. A total of 418 eyes from 222 cases were examined preoperatively, and partly followed up at one week (172 eyes), one month (134 eyes) and three months (51 eyes) after surgery. The horizontal, vertical and total Q values of cornea were calculated from eccentricity measured at the central 6-mm corneal zones respectively. Potential determinants of the change of Q value were analyzed using multiple linear regressions.
RESULTSThe mean Q value was -0.17 +/- 0.13 preoperatively, and 0.99 +/- 0.70, 0.97 +/- 0.66, and 0.86 +/- 0.41 one week, one and three months postoperatively, respectively. One way analysis of variance (ANOVA) demonstrated significant differences between measurements made before surgery and at all postoperative times (at one week, one and three months; all P<0.0001, Bonferroni post hoc), but no significant differences were found among postoperative groups. Significant differences of Q values between horizontal and vertical meridians were found before surgery and at all postoperative times (all P<0.0001). Multiple regression analysis revealed that change of Q value significantly correlated with manifest refraction spherical equivalent (r=0.116, P<0.0001) and axial length (r=0.264, P<0.0001).
CONCLUSIONSOver the study period, the primary changes in Q value occur within 1 week after surgery, and then become slightly decreased and nearly stable. Manifest refraction spherical equivalent and axial length play a significant role in the change of postoperative Q value.
Adolescent ; Adult ; Cornea ; surgery ; Female ; Humans ; Keratomileusis, Laser In Situ ; methods ; Male ; Middle Aged ; Myopia ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult
5.Correlation between expression of forkhead box M1 (FOXM1) and clinicopathological features and prognosis in patients with non-small cell lung cancer (NSCLC).
Yi-qian LIU ; Ren-hua GUO ; Lian-ke LIU ; Wen GAO ; Cheng-jun ZHU ; Juan WEI ; Yong-qian SHU
Chinese Journal of Oncology 2011;33(6):426-430
OBJECTIVETo investigate the expression of forkhead box M1 (FOXM1) and its correlation with clinicopathological features and prognosis in patients with non-small cell lung cancer (NSCLC).
METHODSThe expression of FOXM1 in 68 cases of NSCLC was detected by immunohistochemistry. The FOXM1 expression in 6 tumor tissues (3 cases with negative and 3 cases with positive expression of FOXM1) was analyzed by Western blotting to confirm the immunohistochemical results. The correlation of the expression of FOXM1 with clinicopathalogical features and overall survival of the NSCLC patients was analyzed.
RESULTSThe expression of FOXM1 protein was detected in the nuclei or cytoplasms of the tumor cells. The positive expression rate of FOXM1 was 36.8% (25/68). Western blotting confirmed the immunohistochemical results. The expression level of FOXM1 in advanced stage cancer was significantly higher than that in early stage NSCLC (P = 0.001). The median OS was 23.0 months in patients with negative expression of FOXM1 and 13.0 months in those with positive expression (P = 0.001). Univariate analysis revealed that histological grade, lymph nodes status, TNM stage and FOXM1 expression were significantly associated with prognosis in the NSCLC patients (P < 0.05). The Cox multivariate analysis demonstrated that lymph nodes status, TNM stage and FOXM1 expression were independent poor prognostic factors (P < 0.05).
CONCLUSIONThe expression status of FOXM1 in NSCLC is an independent prognostic factor and negatively correlated with prognosis.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Follow-Up Studies ; Forkhead Box Protein M1 ; Forkhead Transcription Factors ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; Proportional Hazards Models
6.Method performance verification of the clinical chemiluminescence immunoassay
Xiu-Ming ZHANG ; Jun-Hua ZHUANG ; Song-Bai ZHENG ; Jian-Hua XU ; Ji MA ; Lei SUN ; Lian-Ying LIN ; Pei-Feng KE ; Wei-Xiong LIANG
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To establish a method performance verification project and experimental method for the clinical chemiluminescence immunoassay.Methods Referring to CLSI evaluation protocols and pertinent literature,and by combining our actual works,we designed a verification procedure and experimental method.By Using these above,the precision,accuracy,analytical sensitivity,analytical measurement range,clinical reportable range and biotic interval of AFP on the Bayer Centaur 240 chemiluminescence immunoassay system were verificated.Results would be compared with the declaration of the manufacturer or desirable specifications derived from biologic variation.Results The results showed that the between-day inaccuracy on AFP levels at 77.4 ng/ml and 168.0 ng/ml was 5.70% and 4.84% respectively,these were consistent with manufacturer's inaccuracy claimed.The relative bias between the results measured for calibrator at four levels and target value was less 5.0%,and the relative bias between the results measured for EQA control sample at five levels and target value was-3.4% to 11.9%.Lower limit of detection was 1.04 ng/ml,lower slightly manufacturer's analytical sensitivity claimed.Biologic limit of detection was 2.65 ng/ml-3.53 ng/ml,functional sensitivity was 3.53 ng/ml.Analytical measurement range was 3.53-912.00 ng/ml,within manufacturer's liner range claimed.Clinical reportable range was 3.53-182 400.00 ng/ml.Reference interval was 0.6-7.7 ng/ml,within manufacturer' s claimed.Conclusions The main performances of the detection system are accorded with the declaration of the manufacturer.The performance verification procedure and experimental method of our research ars simple and practical,which has important significations for building medical laboratory and laboratory accreditation, improving quality of the chemiluminescence immunoassay.
7.MR diffusion tensor imaging and MRI diffusion-tensor tracking of medial structures of the temporal lobe in the diagnosis of temporal lobe epilepsy
Zhi-Ming LI ; Xian-Yue QUAN ; Li-Lian TAN ; Yong HUANG ; Ke LIU ; Wen LIANG ; Wei WANG
Chinese Journal of Neuromedicine 2011;10(12):1274-1278
Objective To investigate the parameter values and changes of local fiber bundles of the medial structures of the temporal lobe in patients with temporal lobe epilepsy(TLE) with MR diffusion tensor imaging(DTI)and MRI diffusion-tensor tracking(DTT),and evaluate the diagnostic value of DTI in the localization of seizure focus in patients with TLE:Methods Sixteen patients with TLE,admitted to our hospital from December 2010 to February 2011,and 20 healthy volunteers received conventional MRI and DTI.The values of apparent diffusion coefient(ADC),fractional anisotropy(FA)and relative anisotropy(RA) of the bilateral medial structures of the temporal lobe (including the amygdala,hippocampus and cortex)were measured,respectively.The differences of the fiber bundles between the epilepsy patients and the controls were observed with DTT.Results No significant differences in measured values of ADC,FA and RA were noted between the left and right medial structures of temporal lobe in the controls(P>0.05).The values of ADC,FA and RA in the left/right medial structures of temporal lobe in TLE patients had significant differences as compred with those in controls(P< 0.05); the values of ADC showed gradual reduction in the ipsilateral side,contralateral side and controls,successively; and the changes in the hippocampus tail was the most obvious.The values of FA and RA showed gradual increment in the ipsilateral side,contralateral side and controls successively;the changes of parameters values in the amygdala and the hippocampus body were more remarkable than those in the hippocampus tail.Conclusion The DTI can show the micromechanics′ characteristics of the medial structures of the temporal lobe in patients with TLE,which can help the location of seizure focus in patients with TLE.Increasing the understanding of the medial structures of the temporal lobe will help the preoperative integral assessment and improve the surgical effect.
8.Effect of lncRNA MIR17HG on the malignant biological behavior of liver cancer cells by regulating the miR-214-3p/RNF38 signaling axis
Wei-Tao SUN ; Yan-Ke SHI ; Jun-Lian FENG ; Zhi-Fei CHEN ; Cun-Ling ZHANG
Modern Interventional Diagnosis and Treatment in Gastroenterology 2024;29(5):565-571
Objective To explore the effect of long non-coding RNA(lncRNA)microRNA 17-92 cluster host gene(MIR17HG)regulating microRNA(miR)-214-3p/ring finger protein 38(RNF38)signal axis on the malignant biological behavior of liver cancer cells.Methods The cancer tissues and adjacent tissues of 46 patients with liver cancer who underwent surgical resection in our hospital from May 2022 to October 2023 were collected to detect the expression of lncRNA MIR17HG,miR-214-3p and RNF38.HepG2,Bel-7402,SMMC-7721 and HL-7702 cells were cultured in vitro,and the expression of lncRNA MIR17HG,miR-214-3p and RNF38 was compared,Bel-7402 cells were selected for further study,and randomly divided into sh-NC group,sh-MIR17HG group,anti-NC group,anti-miR-214-3p group and Bel-7402 group.The proliferation,apoptosis,invasion and migration of Bel-7402 cells in each group were investigated,the expression of RNF38,caspase-3(caspase-3),B cell lymphoma-2(Bcl-2),matrix metalloproteinase-2(MMP2)and matrix metalloproteinase-9(MMP9)protein was analyzed by western blotting,the relationship between lncRNA MIR17HG and miR-214-3p and the relationship between miR-214-3p and RNF38 were verified by double luciferase.Results The mRNA expression of lncRNA MIR17HG and RNF38 in liver cancer tissues was higher,the mRNA expression of miR-214-3p was lower,and the positive expression rate of RNF38 protein was higher(P<0.05).The expression of lncRNA MIR17HG mRNA,RNF38 mRNA and RNF38 protein in SMMC-7721,HepG2 and Bel-7402 cells was higher than that in HL-7702 cells,and the expression of miR-214-3p mRNA was lower than that in HL-7702 cells(P<0.05).Compared with Bel-7402 group and sh-NC group,the OD450nm value,the number of cloned cells,the number of invasive cells,the number of migrated cells and the expression of RNF38,MMP2,Bcl-2 and MMP9 in sh-MIR17HG group decreased,while the apoptosis rate and the expression of caspase-3 increased(P<0.05).Compared with sh-MIR17HG group and anti-NC group,the OD450nm value,the number of cloned cells,the number of invasive cells,the number of migrated cells and the expression of RNF38,MMP2,Bcl-2 and MMP9 in anti-miR-214-3p group increased,while the apoptosis rate and the expression of caspase-3 decreased(P<0.05).LncRNA MIR17HG and miR-214-3p,and miR-214-3p and RNF38 have targeted relationships respectively.The luciferase activity in miR-214-3p+WT-MIR17HG group was lower than that in miR-NC+WT-MIR17HG group(P<0.05),and the luciferase activity in miR-214-3p+WT-RNF38 group was lower than that in miR-NC+WT-RNF38 group(P<0.05).Conclusion LncRNA MIR17HG may promote the malignant biological behavior of liver cancer cells by regulating the miR-214-3p/RNF38 axis.
9.Changes of motor evoked potentials after spinal cord injuries in rats.
Ke-Wei YU ; Xiao-Jian YE ; Jia-Shun LI ; Wei-Fang RONG ; Lian-Shun JA ; Hai-Long HE
Chinese Journal of Applied Physiology 2002;18(1):14-17
AIMStudy on the relationship between the degraded spinal cord injuries and the changes of the motor evoked potentials (MEP) to prove the diagnosis and prognosis value of MEP.
METHODSAfter injury at T8-T9 cord using modified Allen's weight-drop method, 27 male SD rats were divided randomly into control group (n = 5), group A (50 gcf, n = 8), group B (70 gcf, n = 8) and group C (100 gcf, n = 6). MEPs elicited by monopolar transcortical stimulation were recorded continuously before injury, just after injury, 15 minutes, 1 hour, 3 hours and 6 hours after injury. The rate of the size of the bleeding or necrosis area to the total cord was also calculated.
RESULTSMEP had no significant change in the control group. The amplitude of MEP's early components in group A or group B decreased or even obliterated after SCI, and then partially recovered, while the late components were lost without any recovery signals. All animals in group C showed no MEP waves excepting 2 rats had recovery signals. The size of the cord injuries area increased according to the dropping force and was correlated significantly with the amplitude of the largest peaks of scMEP 1 hour after SCI (r = -0.821).
CONCLUSIONThe scMEP changes after SCI are correlated with the injury forces and the pathological changes in the cord, which indicates that scMEP can be used as an objective index for the cord functional monitoring.
Animals ; Electric Stimulation ; Evoked Potentials, Motor ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; pathology ; physiopathology