1.Optimisation of CUBIC tissue clearing technology based on perfusion methods
Chuan-Hui GONG ; Jia-Yi QIU ; Ke-Xin YIN ; Ji-Ru ZHANG ; Cheng HE ; Ye YUAN ; Guang-Ming LÜ
Acta Anatomica Sinica 2024;55(3):363-370
Objective In order to shorten the transparency time of clear,unobstructed brain imaging cocktails and computational analysis(CUBIC),improve the transparency efficiency,and explore the possibility of applying hydrophilic tissue transparency technique,this study was conducted to optimize the perfusion of CUBIC technique and compare it with four hydrophilic tissue clearing method in terms of tissue transparency effect,transparency time,area change,volume change and adeno-associated virus(AAV)fluorescence retention.Methods Brain,liver,spleen and kidney of 6 adult Institute of Cancer Research(ICR)mice were subjected to clearing treatment by SeeDB,FRUIT,ScaleS and CUBIC method,respectively.The area and gray value of the samples were measured by Image J 1.8.0,and the volume before and after transparency was measured by drainage method to compare the transparency effect,time and size deformation of each group.Perfusion optimization of the CUBIC was performed by improving the perfusion rate with the optimal perfusion dose,each group of the experimental sample size was 6.Fluorescence preservation by different techniques was evaluated by injecting AAV in the motor cortex of 16 adult mice and taking the cervical spinal segments for transparency treatment after four weeks,and the fluorescence photographs were measured by Image J 1.8.0 to measure the mean fluorescent intensity.Results The optimal perfusion rate and dose of CUBIC was 15 ml/min and 200 ml respectively.For transparency ability and speed,the perfusion CUBIC had the lowest mean gray value and took the shortest time,while CUBIC consumed the longest time,and SeeDB,FRUIT,and ScaleS did not show good transparency ability.In terms of area and volume changes,several techniques showed different degrees of expansion after transparency of tissues or organs.In terms of fluorescence retention,perfusion CUBIC showed the best retention of green fluorescent protein(GFP)fluorescence signal,followed by CUBIC,ScaleS,FRUIT,and SeeDB.Conclusion Perfusion CUBIC technique shows the best tissue transparency,the shortest transparency time,and the most AAV fluorescence retention compared with other techniques.
2.Effects of Polybrominated Diphenyl Ethers on the Human Body Exposure in E-Waste Dismantling Region.
Yan ZHOU ; Shao Min LÜ ; Jian Peng XIAO ; Tao LIU ; Wen Jun MA ; Ling Chuan GUO
Journal of Forensic Medicine 2020;36(4):453-460
Objective To discuss the effects of polybrominated diphenyl ethers (PBDEs) exposure in e-waste dismantling region on the human body and provide data support for the identification of environmental health damage to residents in the e-waste dismantling region. Methods Adults in an e-waste dismantling region (exposed group, 54 participants) and a control region (control group, 58 participants) were selected, questionnaires were carried out and blood and urine samples were collected. Blood PBDEs, blood lipids, blood routine, blood lead, urine cadmium, urine chromium and urine nickel were detected. T-test was utilized to compare the differences of PBDEs between the two groups. Multivariate analysis were utilized to compare the differences between the two groups in blood routine indexes. Linear regression was used to analyze the relationship between PBDEs and blood routine. Results Exposure levels of PBDEs were significantly higher in the exposed group (240.00 ng/g, adjusted mass fraction of blood lipids, thereafter) than in the control group (93.00 ng/g, P<0.05). There was no statistical significance in the differences in most blood routine indexes of the two groups ( P>0.05), and their reference values were all within normal ranges. Mean platelet volume, plateletcrit, basophils percentage, absolute value of basophils, and mean corpuscular hemoglobin concentration were higher in the exposed group than in the control group (P<0.05). Platelet distribution widths were lower in the exposed group than in the control group and below the normal reference range (P<0.05). Conclusion PBDEs exposure in e-waste dismantling region tend to change platelet morphology, the number of basophils, and mean corpuscular hemoglobin concentration, and may pose potential health hazards to local residents.
Adult
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China
;
Electronic Waste/analysis*
;
Environmental Monitoring
;
Halogenated Diphenyl Ethers/toxicity*
;
Human Body
;
Humans
3.Impact of blood glucose level on recurrence of liver cancer after laparoscopic surgery
Chuan JIANG ; Chuxiao SHAO ; Jinde ZHU ; Chaoyong TU ; Xiliang Lü ; Qiaomei LIN ; Kun ZHANG
China Journal of Endoscopy 2017;23(8):66-70
Objective To investigate the impact of blood glucose level on the recurrence of liver cancer after laparoscopic surgery. Methods The clinical data of 98 patients with primary hepatocellular carcinoma from January 2012 to January 2015 were retrospectively analyzed. All patients were treated by laparoscopic radical resection of hepatocellular carcinoma. Patients were divided into elevated blood glucose group (n = 23) and control group (n = 75) according to whether the fasting blood glucose was ≥6.1 mmol/L. The recurrence of liver cancer in 1 year and 2 years after operation was compared. The factors influencing the recurrence of liver cancer were analyzed by univariate and multivariate analysis. Results The recurrence rates were 47.82% and 21.33% respectively in the patients with elevated blood glucose and the control group. The recurrence rates were 73.91% and 36.00%respectively in the 2-year postoperative patients with blood glucose and 1 year and 2 years. The recurrence rate was higher than that of the control group, the difference was statistically significant (P < 0.05). Logistic multivariate analysis showed that fasting blood glucose was high, Child-Pugh grade B, intraoperative blood transfusion, lymphatic invasion, high clinical pathology stage, postoperative alpha-fetoprotein (AFP) high, no postoperative adjuvant therapy (P < 0.05). Conclusion The recurrence rate of patients with elevated liver cancer after laparoscopic surgery is high, and fasting blood glucose is high, Child-Pugh grade is B grade, blood transfusion is high, there is lymphatic invasion, high clinical pathology stage after AFP high, no postoperative adjuvant therapy for its postoperative recurrence of risk factors, should strengthen the monitoring of high-risk patients, reduce postoperative recurrence rate.
4.Transperitoneal versus extraperitoneal robot-assisted radical prostatectomy for localized prostate cancer.
Chen-Zhao HUA ; Zhong-Lin CAI ; Wen-Juan LI ; Chuan ZHOU ; Xu-Pan WEI ; Hai-di LÜ ; Feng-Hai ZHOU
National Journal of Andrology 2017;23(6):540-549
Objective:
To compare the clinical effects of transperitoneal (Tp) versus extraperitoneal (Ep) robot-assisted radical prostatectomy (RARP) in the treatment of localized prostate cancer.
METHODS:
We searched PubMed, EMBASE, Web of Science, EBSCO, Cochrane Library, Wanfang, CNKI, and CBM for the articles comparing the clinical effect Tp-RARP with that of Ep-RARP in the treatment of localized prostate cancer published from January 2000 to November 2016. All the articles must meet the inclusion criteria, that is, dealing with at least one of the following aspects: operation time, intraoperative blood loss, postoperative catheterization time, length of bed confinement, perioperative complications, positive surgical margins, bowel-related complications, postoperative anastomotic leakage, and postoperative urinary continence. We subjected the data obtained to statistical analysis using the RevMan5.3 software.
RESULTS:
Two randomized controlled trials and six case-control studies were included in this meta-analysis, involving 451 cases of Tp-RARP and 676 cases of Ep-RARP. Compared with Tp-RARP, Ep-RARP showed significantly shorter operation time (WMD = 21.39, 95% CI: 7.54-35.24, P = 0.002), shorter length of bed confinement (WMD = 0.85, 95% CI: 0.61-1.09, P <0.001), and lower rate of bowel-related complications (RR = 9.74, 95% CI: 3.26-29.07, P <0.001). However, no statistically significant differences were found between the two strategies in intraoperative blood loss (WMD = -8.12, 95% CI: -27.86-11.63, P = 0.42), postoperative catheterization time (WMD = 0.17, 95% CI: -0.55-0.21, P = 0.38), or the rates of perioperative complications (RR = 1.34, 95% CI: -0.97-1.87, P = 0.08), positive surgical margins (RR = 1.24, 95% CI: 0.95-1.61, P = 0.12), anastomotic leakage (RR = 0.98, 95% CI: 0.46-2.10, P = 0.95), urinary continence at 3 months (RR = 0.96, 95% CI: 0.91-1.00, P = 0.05) and urinary continence at 6 months (RR = 1.00, 95% CI: 0.97-1.02, P = 0.82).
CONCLUSIONS
Ep-RARP has the advantages of shorter operation time, shorter length of bed confinement and lower rate of bowel-related complications over Tp-RARP, and therefore may be a better option for the treatment of localized prostate cancer. However, more multi-centered randomized controlled clinical trials are needed for further evaluation of these two approaches.
Blood Loss, Surgical
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Case-Control Studies
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Humans
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Male
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Margins of Excision
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Operative Time
;
Postoperative Complications
;
Prostatectomy
;
adverse effects
;
methods
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Prostatic Neoplasms
;
pathology
;
surgery
;
Randomized Controlled Trials as Topic
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Robotic Surgical Procedures
;
adverse effects
;
methods
;
Treatment Outcome
5.MiR-124 suppresses the proliferation of human prostate cancer PC3 cells by targeting PKM2.
Lei LÜ ; Jing-Dong YUAN ; Zuo-Liang CAO ; Tao HUANG ; Chuan-Hua ZHANG ; Liang WANG ; Fu-Qing ZENG
National Journal of Andrology 2014;20(6):495-499
OBJECTIVETo explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells.
METHODSLuciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay.
RESULTSThe expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05).
CONCLUSIONmiR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.
Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Hormones ; genetics ; metabolism ; Transfection
6.Effects of Helicobacter pylori on the expression of catalytic subunit of the DNA-dependent protein kinase and Ku70/Ku80 heterodimer in gastric mucosa epithelial cells in vivo and in vitro
Wei LI ; Chuan XIE ; Zhen YANG ; Nonghua Lü
Chinese Journal of Digestion 2013;33(10):675-679
Objective To explore the effects of Helicobacter pylori (H.pylori) on the expression of catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs) and Ku70/Ku80 heterodimer in gastric mucosa epithelial cells in vivo and in vitro.Methods After treated with H.pylori for one,three,six,12 and 24 hours,the expressions of DNA-PKcs and Ku70/Ku80 heterodimer in gastric epithelial cells (GES) 1 and gastric adenocarinoma cells (AGS) were detected by Western blot.Mongolian gerbils were gavaged with H.pylori,and were sacrificed after infected for six and 12 months.The gastric mucosa tissues were taken for immunohistochemistry to detect the expressions of DNA-PKcs and Ku70/Ku80 heterodimer at protein level.The data were analyzed by t test and chi-square test.Results After H.pylori infection for one hour,the relative quantity of the expression of DNA-PKcs in GES-1 was 1.16±0.09,which was higher than that of non infected group (1.04±0.31) and the difference was statistically significant (t=4.67,P<0.05).After infected by H.pylori for one,three,six,12 and 24 hours,the relative quantities of the expressions of Ku70/Ku80 heterodimer in GES-1 were 1.58±0.32,1.84±0.40,1.97±0.35,3.72±1.42 and 3.74±1.56,respectively,all were higher than that of non infected group (1.24±0.31) and the differences were statistically significant (t=3.57,4.20,5.03,8.11 and 8.14,all P<0.05).The relative quantities of the expressions of Ku70/Ku80 heterodimer in AGS were 4.69 ± 0.87,3.67 ± 0.67,2.41±0.24,1.35±0.35 and 1.32±0.10 after H.pylori infected for one,three,six,12 and 24 hours,respectively,all were lower than that of no H.pylori infected group (4.84 ± 0.76) and the differences were statistically significant (t=34.13,27.68,19.81,4.47 and 5.69,all P<0.05).In Mongolian gerbil models,DNA-PKcs did not express in H.pylori negative group (0/25),the total positive rate of H.pylori infected group was 98.1% (53/54),the difference between the two groups was statistically significant (x2 =74.55,P<0.01).The total positive rate of Ku70/Ku80 heterodimer in H.pylori negative group was 92.0% (23/25) and in H.pylori infected group was 68.5% (37/54),the difference between the two groups was statistically significant (x2=5.16,P<0.05).Conclusion H.pylori infection affected cellular DNA damage repair through changing the expression of DNA-PKcs and Ku70/Ku80 heterodimer in gastric mucosa in vivo and in vitro,which may cause gastric mucosal lesions.
7.Echinococcus multilocularis infection leads to increased expression of proliferation and anti-apoptosis factors in liver cells in BALB/c mice.
Chuan-shan ZHANG ; Jun-hua WANG ; Guo-dong LÜ ; Liang LI ; Hao WEN ; Ren-yong LIN
Chinese Journal of Hepatology 2013;21(3):228-233
OBJECTIVETo investigate the effects of Echinococcus multilocularis on host liver cell proliferation in vivo using a BALB/c mouse alveolar hydatid infection model.
METHODSSixty-five 8-10-week-old female BALB/c mice were randomly divided into an experimental group (n = 40) and a control group (n = 25) and administered an abdominal injection into the left liver lobe of E. multilocularis protoscolices in saline solution or saline solution alone, respectively. At post-injection day 2, 8, 30, 60, and 90, liver samples were collected for analysis of lesions and lesion-adjacent tissue by hematoxylin-eosin staining and differential expression of proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin A, and cyclin B1 by immunohistochemical staining. The significance of intergroup differences was assessed by Student's t-test.
RESULTSThe control group showed normal liver histology at all time points. The experimental group developed E. multilocularis lesions that showed increased severity of pathological features, such as inflammatory cell invasion, steatosis and fibrous connective tissue hyperplasia, over time. At post-injection days 2 and 8, enlarged, binuclear and apocyte hepatocytes were observed close to the lesions. At post-injection days 30, 60, and 90, the number of hepatocytes expressing PCNA progressively increased in the experimental group, and the numbers were significantly higher than in the control group (7.01 +/- 1.89 vs. 1.03 +/- 0.52, 8.41 +/- 2.80 vs. 0.93 +/- 0.31, and 13.4 +/- 4.43 vs. 1.07 +/- 0.94; all P < 0.05). The same progressively increasing trend was seen in the number of hepatocytes expressing CyclinD1, but was only significantly different from controls at post-injection days 30 and 60 (6.73 +/- 2.52 vs. 0.48 +/- 0.43 and 8.22 +/- 3.09 vs. 0.55 +/- 0.34; both P < 0.05). In contrast, the number of hepatocytes expressing cyclin A was significantly increased at post-injection day 30 and then showed a decreasing trend at days 60 and 90, although the numbers of expressing cells remained significantly higher than control levels at all time points (7.75 +/- 3.05 vs. 0.69 +/- 0.36, 3.42 +/- 1.80 vs. 1.14 +/- 0.42, and 3.03 +/- 1.50 vs. 0.69 +/- 0.31; all P < 0.05). The number of hepatocytes expressing CyclinB1 in the experimental group was less robust than the other cyclins (with a general temporal trend of increase followed by decrease), but the differential expression was not significantly different from the control levels at any time point.
CONCLUSIONE. multilocularis infection may promote the expression of host factors related to proliferation and anti-apoptosis in liver. This pathogen-mediated modulation of host cell-survival mechanisms may provide a rationale explanation for the clinical observations of hepatomegaly and the unexpected survival of alveolar echinococcosis patients following major hepatic resection.
Animals ; Apoptosis ; Cell Cycle ; Cell Proliferation ; Echinococcosis ; pathology ; Echinococcus multilocularis ; Female ; Hepatocytes ; cytology ; pathology ; Liver ; pathology ; Mice ; Mice, Inbred BALB C
8.Silencing pyruvate kinase M2 sensitizes human prostate cancer PC3 cells to gambogic acid-induced apoptosis.
Lei LÜ ; Liang WANG ; Guo-Song JIANG ; Chuan-Hua ZHANG ; Fu-Qing ZENG
National Journal of Andrology 2013;19(2):102-106
OBJECTIVETo study the effect of silencing pyruvate kinase M2 (PKM2) on gambogic acid (GA)-induced apoptosis of human prostate cancer PC3 cells.
METHODSThree specific PKM2 siRNAs and one negative control siRNA (si-NC) were transfected into PC3 cells. The silencing effect of PKM2 siRNAs was determined by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, and the effects of PKM2 siRNA on the vitality and apoptosis of GA-stimulated PC3 cells detected by MTT and AO/EB double staining, respectively. The mRNA and protein levels of c-myc and cyclin D1 were analyzed by qRT-PCR and Western blot, respectively.
RESULTSAll the 3 PKM2 siRNAs effectively reduced the mRNA and protein expressions of PKM2, and PKM2 siRNA-1 exhibited the strongest silencing effect. At 24 h after transfection, the expression levels of PKM2 mRNA and protein were reduced by 70% and 85%, respectively (P < 0.05). Twenty-four hours of treatment with GA (0.5 micromol/L) following transfection with PKM2 siRNA-1 inhibited the vitality of the PC3 cells by 68%, increased their apoptosis, and significantly down-regulated the mRNA and protein levels of c-myc (50% and 35%) and cyclin D1 (60% and 20%) (P < 0.05).
CONCLUSIONInhibition of PKM2 sensitized PC3 cells to GA-induced apoptosis, suggesting that PKM2 may be a potential therapeutic target for sensitizing human prostate cancer to GA.
Apoptosis ; drug effects ; Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA Interference ; RNA, Small Interfering ; Thyroid Hormones ; genetics ; metabolism ; Xanthones ; pharmacology
9.The changes of peripheral blood lymphocyte subsets in patients with acute hydrogen chloride gas poisoning.
Ya-li LÜ ; Chuan-jun CHEN ; Bi-xia LIANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(5):375-376
OBJECTIVETo explore the changes of peripheral blood lymphocyte subsets in patients with acute hydrogen chloride gas poisoning.
METHODSWhen the patients were admitted or on the secondary day, the percentages of total T-cell lymphocyte subsets (CD(3)(+)CD(19)(-)), CD(4)(+)T cells (CD(3)(+)CD(4)(+)), CD(8)(+)T cells (CD(3)(+)CD(8)(+)), B cells (CD(3)(-)CD(19)(+)) and NK cells (CD(3)(-)CD(16)(+)CD(56)(+)), and the ration of CD(4)(+)/CD(8)(+) in 37 cases with acute hydrogen chloride gas poisoning and 49 healthy controls were detected with flow cytometer.
RESULTSThe total T-cell percentage and total CD(4)(+)T cell percentage in 37 cases were significantly lower than those in 47 controls (P < 0.05). The percentages of NK cells and B lymphocytes in 37 cases significantly increased, as compared with controls (P < 0.05). The ration of CD(4)(+)/CD(8)(+) in 37 cases significantly decreased, as compared with controls (P < 0.05).
CONCLUSIONThe lymphocyte subsets in the patients with acute hydrogen chloride gas poisoning changed, which could influence the immune function of patients.
Adolescent ; Adult ; Case-Control Studies ; Female ; Gas Poisoning ; blood ; Humans ; Hydrochloric Acid ; poisoning ; Lymphocyte Count ; Lymphocyte Subsets ; cytology ; Male ; Middle Aged ; Young Adult
10.Estimation of postmortem interval by detecting thickness of cornea using ultrasonic method.
Guo-Li LÜ ; Fu-Xue JIANG ; Xin-Shu XU ; Yong-Jun JIANG ; Zhi-Gang LI ; Xin WANG ; He SHI ; Li-Cong YU ; Chuan-Chao XU
Journal of Forensic Medicine 2012;28(2):89-91
OBJECTIVE:
To explore the postmortem changes of cornea thickness measured by ultrasonic pachymetry.
METHODS:
Eleven rabbits were randomly divided into two groups: one group with intact corneal epithelium and another group without intact corneal epithelium. In the later group, the corneal epithelium of the rabbit was scraped using mechanical elimination method. The corneal thickness was monitored continuously by ultrasonic pachymetry at several postmortem interval points in rabbits of the two groups. The changes of corneal thickness and postmortem interval were explored by relative regression analysis.
RESULTS:
The thickness of the cornea showed a strong non-linear correlation with the postmortem interval in the group with intact corneal epithelium. The group with intact corneal epithelium showed the correlation coefficient 0.922 and the group without intact corneal epithelium showed the correlation coefficient 0.822, respectively.
CONCLUSION
The corneal thickness measured by ultrasonic pachymetry shows a potential value for estimating early postmortem interval. The intact corneal epithelium is a crucial factor for the measurement of cornea thickness by ultrasonic pachymetry.
Animals
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Cornea/pathology*
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Corneal Topography/methods*
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Epithelium, Corneal/ultrastructure*
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Female
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Forensic Pathology/methods*
;
Male
;
Postmortem Changes
;
Rabbits
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Regression Analysis
;
Reproducibility of Results
;
Time Factors
;
Ultrasonography

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