Effects of Acupuncture Based on Acupoint Selection of Myofascial Meridians Perspective on Regeneration of Nerve Cells and Recovery of Motor Function in Rats with Cerebral Palsy / 中国康复理论与实践

@#Objective To investigate the effects of acupuncture based on acupoint selection of myofascial meridians perspective on the regeneration of nerve cells and recovery of motor function in rats with cerebral palsy. Methods 20 healthy 21-day-old male Sprague-Dawley rats were divided into control group, acupuncture groups 1 (receiving traditional acupuncture) and 2 (receiving acupuncture based on acupoint selection of myofascial meridians perspective), and sham group. The control group and the acupuncture groups were subjected to ligation of left carotid artery (ischemia) and then put into a hypoxic environment. They were assessed with Basso-Beattie-Bresnahan (BBB) Scale and inclined plane test 3 d, 7 d and 14 d after the beginning of acupuncture. And 5-bromo-2'-deoxyuridine (BrdU) was used to label Sphase cells in the left striaturn and motor cortex in each group 14 d after the beginning of acupuncture. Results The BBB scores improved more in the acupuncture group 2 than in the acupuncture group 1 and the control group 7 d, 14 d after the beginning of acupuncture (P<0.01), as well as the score of inclined plane test 3 d, 7 d, 14 d after acupuncture (P<0.05). BrdU was more in the left motor cortex in the acupuncture group 2 than in the acupuncture group 1 (P<0.05). Conclusion Acupuncture based on acupoint selection of myofascial meridians perspective can promote the regeneration of nerve cells and recovery of motor function in rats with cerebral palsy, that is more effective than traditional acupuncture.

Effects of rehabilitation on the regeneration of nerve cells after experimental cerebral hemorrhage in rats / 中华物理医学与康复杂志

Objective To study the effects of rehabilitation training on the regeneration of nerve cells in rats after intracerebral hemorrhage (ICH).Methods A total of 75 male SD rats were randomized into a training group,a control group and a sham operated group,25 rats/group.The ICH models were induced by stereotactical injection of collagenase type Ⅶ into the globus pallidus.The training group was trained with grasp,balancing and rotating exercise every day,the control group was restricted to their cages,and the sham operated group received normal saline injections.Each group was further subdivided into 1,4,7,14 and 28 day subgroups.Neurological function was measured in each group.Bromodeoxyuridine (BrdU) was used to label S phase cells,immunohistochemical single and double staining with antibodies against BrdU,microtubal-associated protein (MAP) and neuronal nuclei (NeuN) were used to determine neuronal proliferation,migration and differentiation in the subventricular zone ( SVZ ) and subgranular zone (SGZ) in the training and control groups.Results The motor function scores of the animals in the rehabilitation group were significantly lower than those of the control group.Proliferative BrdU + cells of the SVZ and SGZ in the control group rats were clearly less than those in the rehabilitation training rats at all time points.The results of the immunohistochemical double staining indicated that one week after ICH BrdU +/MAP + cells in the SVZ had increased significantly in the training group compared to the control group,and then decreased two weeks later.At the same time,BrdU +/MAP cells were found in the striatal boundary on the hemorrhage side,in numbers up to 8 times that in the control group.In the rehabilitation group striatal neuron differentiation on the hemorrhage side was 2 to 3 times that in the control group.Conclusion Rehabilitative training can enhance nerve cell proliferation,regeneration and neuron migration after ICH.

TPA and BrdU promote CD133 expression in nasopharyngeal carcinoma cells / 肿瘤

Objective:To explore whether tumor-inducing agent 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and bromodeoxyuridine (BrdU) affect CD133 expression in nasopharyngeal carcinoma (NPC) 5-8F cells. Methods:NPC cell line 5-8F was treated with single TPA, single BrdU, or TPA plus BrdU, respectively. CD133 mRNA and protein expression levels were detected by real-time fluorogentic quantitative PCR and Western blotting, respectively. Flow cytometry was used to separate CD133-positive cells and determine their levels. Boyden chamber test was used to measure the invasion capability of the cells. Results:Compared with untreated group, CD133 mRNA levels were increased in single BrdU group and BrdU plus TPA group (P=0.037 and 0.003, respectively), and decreased in single TPA group. Western blotting indicated that the expressions of CD133 protein was increased in all the three treated groups, and FCM showed that the quantity of CD133-positive cells also increased. The invasion capability was enhanced, especially in BrdU plus TPA group. Conclusion:Both TPA and BrdU increased CD133 expression in NPC.The effects of TPA and BrdU are synergestic.

In vitro bromodeoxyuridine-labeled goat bone marrow mesenchymal stem cells / 中华创伤骨科杂志

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.

Kidney Tubular Cell Regeneration Starts in the Deep Cortex after Ischemia / 대한신장학회지

PURPOSE: In kidneys exposed to ischemia/reperfusion (I/R), the periodic and regional changes of loss and restoration of tubular epithelial cells and the influence of these processes for renal function remain to be defined. We investigated the loss and regeneration of tubular cells in each nephron segment at various times after I/R. METHODS: Mice were subjected to 30 min of bilateral renal ischemia and were administered 5-bromo-2'-deoxyuridine (BrdU) 20 hours before harvest kidneys. The numbers of tubular cell nuclei, BrdUincorporating cells and proliferative cell nuclear antigen (PCNA)-positive cells were analyzed by PASstaining and immunohistochemistry. RESULTS: Thirty minutes of ischemia induced loss of tubular epithelial cells in the outer stripe of the outer medulla. The loss of tubular epithelial cells peaked 24 hours after ischemia. After the maximum decrease, recovery of number of tubular epithelial cells was observed from 3 days after I/R in the outer medulla and from 5 days in the cortex. The tubular cell numbers were inversely correlated with the changes in concentrations of plasma creatinine and BUN by Pearson correlation, indicating that the decrease and increase of tubular epithelial cell numbers reflect functional failure and recovery, respectively. Cell proliferation as determined by BrdU-incorporating appeared in the deep cortex from 3 days after ischemia. CONCLUSION: The recovery of renal function was found to significantly correlate with the restoration of tubular cells. Furthermore, the regeneration of tubular cells started in the tubules of the deep cortex, suggesting that it may be a great proliferative cell niche.

Mesenchymal stem cells promote proliferation of endogenous neural stem cells and survival of newborn cells in a rat stroke model

Mesenchymal stem cells (MSCs) secrete bioactive factors that exert diverse responses in vivo. In the present study, we explored mechanism how MSCs may lead to higher functional recovery in the animal stroke model. Bone marrow-derived MSCs were transplanted into the brain parenchyma 3 days after induction of stroke by occluding middle cerebral artery for 2 h. Stoke induced proliferation of resident neural stem cells in subventricular zone. However, most of new born cells underwent cell death and had a limited impact on functional recovery after stroke. Transplantation of MSCs enhanced proliferation of endogenous neural stem cells while suppressing the cell death of newly generated cells. Thereby, newborn cells migrated toward ischemic territory and differentiated in ischemic boundaries into doublecortin+ neuroblasts at higher rates in animals with MSCs compared to control group. The present study indicates that therapeutic effects of MSCs are at least partly ascribed to dual functions of MSCs by enhancing endogenous neurogenesis and protecting newborn cells from deleterious environment. The results reinforce the prospects of clinical application using MSCs in the treatment of neurological disorders.

Study of Survival and Migration of Schwann Cells Transplanted to Central Nervous System for A Long-term / 中国康复理论与实践

@#ObjectiveTo investigate Schwann cells whether survival and migration after transplanted to central nervous system for a long-term.MethodsThe Schwann cells of rat were expended in vitro, the part of them were labeled with 5'-bromodeoxyuridine (BrdU) and transplanted to rat's middle brain injured by electric needle stimulus, the others were labeled with Hoechst 33342, seeded to PLGA scaffold, and transplanted to rat's transected spinal cord. 8 and 11 months later, rat brain and spinal cord were taken out respectively, examined by BrdU immunohistochemistry and fluorescence microscope.ResultsBrdU positive cells could be seen after 8 months and migrated toward cerebral cortex. Hoechst 33342 positive cells could be identified in scaffold and transected spinal cord after 11 months under fluorescence microscope.ConclusionGrafted Schwann cells can survive in central nervous system for a long-term and migrate toward distance.

Chronic mild stress decreases survival, but not proliferation, of new-born cells in adult rat hippocampus

New-born cells continue to proliferate and survive to become mature granule cells in adult rat hippocampus. Although this process, known as neurogenesis, is inhibited by acute stress, it is not clear whether chronic stress affects neurogenesis. To determine whether chronic mild stress (CMS) influences neurogenesis in the adult rat hippocampus, male Sprague-Dawley rats were exposed to CMS and administered bromodeoxyuridine (BrdU) before or after CMS to observe the survival/differentiation or proliferation of new-born cells, respectively. In addition, we measured brain-derived neurotrophic factor (BDNF) mRNA in the granule cell layer (GCL) of the hippocampus, because BDNF is known to play an important role in the survival of new-born cells. CMS significantly decreased the survival of newborn cells in the GCL, but did not influence the proliferation or differentiation of new-born cells. CMS did not affect the proliferation and survival of new-born cells in the hilus. In addition, CMS did not change BDNF mRNA levels in the GCL. These results demonstrate that CMS reduces the survival of new-born cells but not of their proliferation, suggesting that repeated mild stress could influence a part of neurogenesis, but not the whole part of neurogenesis. These results raise the possibility that the survival of new-born cells may be suppressed in the presence of normal BDNF mRNA levels in GCL.

Protective Effect of Fastigial Nucleus Electric Stimulation on Brain of Rats with Hypoxic-Ischemic Brain Damage / 实用儿科临床杂志

Objective To explore the protective effect and the mechanism of fastigial nucleus electric stimulation on brain of rats with hypoxic-ischemic brain damage(HIBD).Methods Ninety rats were randomly divided into 3 groups:sham operated group(n=30),model group(n=30)and electric stimulation group(n=30),every group was divided into group A(n=10),group B(n=10)and group C(n=10)again.The models of perinatal HIBD rats were prepared by ligation of left common carotid artery with a temporary systemic hypoxia(N2O2 was 928)for 2 hours.Electric stimulation group was used electric stimulation for 20 minutes,2 times everyday after surgery.The sham operated group and model group was not used electric stimulation but catched to fix in corresponding period.All of the group A would be injected bromodeoxyuridine(BrdU)to enterocoelia before 8 hours when the rats would be killed and the group B would be not injected it.The rats of the group A and B would be killed and got the brain tissue after cardiac perfusion 7 days later,then,consecutively coronal slice.The changes of BrdU and nestin levels in brain were observed by immunohistochemistry staining assay.And the study and memory ability of all the group C would be tested by maze test after 28 days.The brain tissue would be tested by hematoxylin and eosin stain at the same time.All of the data would be described and analyzed by SPSS 13.0.More than a few means would be compared by One-Way analysis of variance and the t-test between 2 groups would be used.Results The BrdU and nestin levels of the model group were lower than the sham operated group(Pa

Transplantation of Human Umbilical Cord Blood Improves Neurological Outcomes in the Rats after Traumatic Spinal Cord Injury

OBJECTIVE: Recent advances in stem cell biology make it possible to induce the regeneration of injured axons and to replace lost cells in the injured spinal cord. It has been found that stem cells in human cord blood differentiate into mature neurons and glial cells both in vitro and in vivo. These findings suggest that human umbilical cord blood cells(HUCBs) can be used as therapeutic donor cells in cases of spinal cord injury. METHODS: To attempt the repair an injured cord following spinal cord injury(SCI), we transplanted HUCBs into contused spinal cords. This was found to promote a long-term improvement in neurologic function relative to a lesion-control group. HUCBs were cultured in vitro for 7 days. Bromodeoxyuridine(BrdU) was added to the media to allow the BrdU to integrate into dividing cells. Cultured HUCBs(2x106 cells) were then injected into the injury epicenter 7 days after SCI. The Basso-Beattie-Bresnahan(BBB) locomotor rating system was used to score functional improvement in HUCBs transplanted rats. Immunohistochemical staining for neurofilament, macrotubule associated protein 2(MAP-2), glial fibrillary acidic protein(GFAP), and nestin was performed. RESULTS: Immunohistochemical analysis 5 weeks after SCI showed that gliogenesis of the transplanted donor HUCBs had occurred within the adult rat spinal cord. These donor-derived astrocyte-like cells extended their processes into the host tissues and integrated well. HUCBs derived neurons(neurofilament, MAP-2) and nestin expressing cells were also detected. Behavior analysis using BBB rating scores showed that functional improvement was greater in transplanted rats than in non-treated rats. CONCLUSION: HUCBs are one of the potential sources for transplantation material for the treatment of SCI.

Effect of noggin gene on proliferation of neural precursor cells from the adult rat hippocampus / 第三军医大学学报

Objective To investigate the effect of noggin on BrdU labeled cells in the adult rat hippocampus. Methods The expressions of noggin and bone morphogenetic protein 4 (BMP4) in rat hippocampus were detected using in situ hybridization histochemistry (ISHH) and reverse transcription polymerase chain reaction (RT PCR). By using antisense technique combined with bromodeoxyuridine!(BrdU) labeling, the effect of noggin on hippocampal neurogenesis in adult rats was explored. Results The number of noggin mRNA positive cells in the adult rat hippocampus decreased significantly after treatment with antisense noggin but no change was found in the number of BMP4 mRNA positive cells. In addition, the number of BrdU labeled cells decreased significantly in the adult rat hippocampus after treatment with antisense noggin, but the sense noggin had no such effect. Conclusion Noggin can promote proliferation of neural precursor cells in adult rat hippocampus.

Effects of ginsenoside Rgl on the proliferation of neural stem cells in rats with focal cerebral ischemia / 解放军医学杂志

Objective To investigate the influence of ginsenoside Rgl on the proliferation of neural stem cells in adult rats with fo- cal cerebral ischemia,and to explore the possible mechanism of ginsenoside Rg1 in brain protection and anti-aging action.Methods The animal model of left middle cerebral artery occlusion(MCAO)was reproduced in adult male Wistar rats.The thymidine analog bromode- oxyuridine(BrdU)was administered intraperitoneally(50mg/kg,Sigma,USA)every four hours for four times before executing the ani- mals to label the proliferating cells.By the employment of immunohistochemical single staining and double-immunofluorescence technique, the number of BrdU immunoreacted nuclei in the subventricular zone(SVZ),the number of nestin positive cells in the subgranular zone (SGZ)as well as the nestin/BrdU double-labeled positive cells were counted.The effect of ginsenoside Rg1 on the immunoreactivity for BrdU,nestin and nestin/BrdU at 1d,3d,7d and 14d after focal cerebral ischemia were also observed to compare with that of controls,Re- sults The immunoreacted cells of BrdU,nestin and nestin/BrdU were seen in SVZ and SGZ in the hippocampus following focal cerebral ischemia.The number of BrdU,nestin-positive cells and the double-labeled positive cells all reached the peak on 7d and decreased on 14d after MCAO,and the expressions markedly increased after the rats were given ginsenoside Rg1,compared with the control(P

FLOW CYTOMETRIC MEASUREMENTS OF CELL CYCLE KINETICS BY BROMODEOXYURIDINE(BRDU)/DNA BIP- ARAMETER ANALYSIS AND THEIR PROGNOSTIC VALUE IN ACUTE LEUKEMIA / 解放军医学杂志

To study cell cycle kinetics of acute leukemia cells and to find out its relationship with clinical treatment outcomes, flow cyto-metric measurements of cell cycle kinetics, especially percentage of S-phase cells (S%) and duration of S-phase(Ts), were made in bone marrow cells of 39 patients with acute leukemia, including 26 cases of ANLL and 13 cases of ALL. Among them 31 cases were at diagnosis and 8 were in relapes. Bromodeoxyuridine(BrdU) / DNA biparameter analysis was used, in which cells were stained with propidium iodide(PI) to estimate total cellular UNA content and a monoclonal antibody against BrdU as a probe for BrdU incorporated into DNA. The results showed that the mean S% of bone marrow cells was significantly lower than that of normal controls, while there was no significant difference between S% of patients at diagnosis and of those in relapse. However, the mean Ts was not significantly different in patients and in normal subjects, but Ts of patients in relapse was significantly shorter than that of the patients at diagnosis and that of normal controls. It is suggested that the proliferation rate of leukemic cells is lower than normal. Shorter Ts indicates faster regrowth of leukemic cells, which is closely related to relapse.

EFFECT OF SENSORY NEUROPEPTIDE ON MIGRATION OF EPIDERMAL STEM CELLS IN WOUND REPAIR / 解放军医学杂志

Objective To study the effect of neuropeptide on migration of epidermal stem cells in wound repair. Methods 90 new born rats (3～4d old) were randomly divided into three groups: SP group, capsaicin group, and normal group. BrdU labeling, combined with specific protein markers of epidermal stem cells, K19 and ?1 integrin, were used to identify epidermal stem cells. The migration of epidermal stem cells was observed in full thickness skin wound on back on 21 days after injury. Substance P was applied to the skin wound after injury in SP group. The status of migration of epidermal stem cells in SP group was compared with that in capsaicin group, in which capsaicin was injected to destroy sensory neuron before skin injury, and with normal group, in which the skin wound was not treated with any medication. Results Wounds of rats in SP group were healed 18 days after injury. It was shortened by three days compared with normal group. Only 25.54% of wound area was healed in capsaicin group on day 18. There were many epidermal stem cells in the edge of the wound and granulation tissue in Group SP. Only a small number of epidermal stem cells were seen in capsaicin group, where SP release was blocked by chemical destruction of sensory neurons, in the wound edge, but not in granulation tissue. The amount of epidermal stem cells as seen in the wound edge, but not in granulation tissue, in normal group was less than in the SP group but more than in capsaicin group. Conclusions SP obviously promotes wound healing and shortens the healing period. SP can induce epidermal stem cells to migrate into the skin wound edge and granulation tissue.

Stem cells marked with bromodeoxyuridine and telomerase reverse transcriptase in hyperoxia lung injury in neonatal rats / 中华围产医学杂志

Objective To investigate the characteristics of stem cells marked with bromode oxyuridine (Brdu) and telomerase reverse transcriptase (TERT) in lung tissue, as well as its effects on pulmonary development and injury-repair. Methods A model of hyperoxia in neonatal rats was established by exposed to 95% O2 for 7 days. Before executing rats, Brdu was injected peritoneally, then Brdu and TERT positive cells were detected by immunohistochemistry. Results (1)The positive staining cells of Brdu with large nuclear located in septa and submucose of series bronchia, scattering in epithelium of bronchia, and the number of positive cells were less. The positive staining cells of TERT located in the septa and alveolar walls of peripulmonary tissue and the number of which was less than that of Brdu. (2)The positive cells of SPC located in septa and alveolar walls. Staining with Brdu and TERT, small number of positive cells was observed. (3) In hyperoxia and normal oxygen group, integral of expression of Brdu, TRET and SPC had no differences. But integral of expression of Brdu in whatever hyperoxia (1. 61?0. 83) or normal oxygen group (1. 43?0. 85) were higher than TRET and SPC (P

Expression of Osteocalcin and Transglutaminase C during Fracture Healing and Distraction Osteogenesis in Rat's Tibia / 대한정형외과학회잡지

Incorporation of bromodeoxyuridine(BrdU) and expression of osteocalcin and transglutaminase C(TGase C) during fracture healing and distraction osteogenesis were investigated in the rat with immunohistochemical studies. Transverse osteotomy was made at the proximal tibia. Bilateral dynamic mini-fixator was applied to immobilize the fracture and also to lengthen the leg. Distraction was started, at the rate of 0.25 mm twice daily, from the 4th operative day and continued for 7 days. Animals were killed for immunohistochemical studies on the 1st, 3rd, 5th, 7th, 14th, 28th, 42nd, 56th, and 84th day after osteotomy or distraction. Longitudinal histologic sections of the healing bone were stained with monoclonal antibodies against BrdU, osteocalcin, and TGase C. Radiologically, complete fracture healing was achieved in 6 weeks after osteotomy, while neo-osteogenesis was successfully achieved in the distracted gap in 7 weeks after the completion of distraction, During active healing stage of the fracture and distraction osteogenesis, BrdU was mainly expressed in the perisoteal and endosteal osteoprogenitor cells while osteocalcin was expressed in the proliferating osteoprogenitor cells, osteoblast, osteocyte, osteoid matrix, and chondrocyte. The expression of BrdU and osteocalcin in the mesenchymal cells from the surrounding soft tissues around the osteotomy site was negligible. At the site of enchondral bone formation, TGase C was expressed in the cytomplasm of more centrally located and matured chondrocytes, while oseocalcin was mainly expressed in the cytoplasm of peripherally located chondrocyte. These findings may suggest that osteocalcin participates in early phase of enchondral bone formation, while TGase C in the late phase, suggesting the role of TGase C in matrix stabilization. At the site of intramern-branous bone formation, the expression of TGase C was weakly positive in both osteoprogenitor cell and osteoblast. The reason of the difference in the expression of TGase C between the enchondral bone formation and intrarnembranous bone formation should be further investigated. Fracture healing and distraction osteogenesis was predominantly induced by intramembranous ossification rather than enchondral ossification. Periosteal osteoprogenitor cells appeared to initiate and to lead bone formation after osteotomy and distraction. Active proliferation and differentiation of osteoprogenitor cell ocurred during entire periods of distraction. Also, active osteoid matrix formation and mineralization was started from the 5th day of distraction and continued thereafter for further 4 weeks after completion of the lengthening. These findings indicate that preservation of the periosteum is essential to achieve successful fracture healing and distraction osteogenesis.

Cell Kinetic Evaluation of Bromodeoxyuridine and Proliferating Cell Nuclear Antigen Expressed Cells in Experimental Bladder Tumorigenesis Using Color-Image Analysis / 대한비뇨기과학회지

A study was carried out to define the role of color-image analysis in evaluation of immunohistochemically labeled cells using anti-BrdU monoclonal antibody and anti-PCNA/cyclin monoclonal antibody, and to clarify the characteristic of nodular or papillary hyperplasia lesion using color-image analysis. Sequential cellular changes(normal, simple hyperplasia, nodular or papillary hyperplasia, and transitional cell carcinoma-Ta, T1) were observed in Fisher 344 rat urinary bladder depending on the duration of BBN administration. The mean BrdU labeling index obtained using color-image analysis and visual analysis showed statistically significant difference between normal and simple hyperplasia, and nodular or papillary hyperplasia and bladder tumor, but no difference between simple hyperplasia and nodular or papillary hyperplasia. The mean PCNA labeling index obtained by color-image analysis and visual analysis in normal, simple hyperplasia, nodular or papillary hyperplasia and bladder tumor. showed statistically significant difference in each histological group. But coefficient of variation of mean BrdU labeling index obtained by color-image analysis was higher than that by visual analysis only in normal bladder mucosa, and was lower in simple hyperplasia, nodular. or papillary hyperplasia, and bladder tumor. Coefficients of variation of mean PCNA labeling index obtained by color-image analysis were lower than those using visual analysis in all lesions. Higher BrdU and PCNA labeling indices indicated greater biological malignant potential,and mean labeling index in each histological group obtained by both color-image and visual analyses progressively increased with tumorigenesis. The coefficients of variation with color-image analysis were lower than those with visual analysis in general, therefore. the evaluation of labeling indices obtained using color-image analysis was more reliable than that obtained using visual analysis. Nodular or papillary hyperplasia showed rather a premalignant lesion in dynamic process than the previous concept of two separated entities, reversible or irreversible lesion, when evaluated more reliably by color-image analysis. The data suggest that cell kinetic evaluation using color-image analysis provide more reliable information to evaluate immunohistochemical staining in terms of quantitation and reproducibility and play an important role in predicting proliferative activity and malignant potential for bladder tumor.

Comparison of brain tumor growth kinetics by proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU) labeling

The bromodeoxyuridine (BrdU) labeling study provides valuable cell kinetic information for individual tumors that could suggest the prognosis of each patient who had a tumor. Recently, a monoclonal antibody against the proliferating cell nuclear antigen (PCNA or cyclin), a nuclear protein expressed in proliferating cells, was developed which could be used on formalin fixed, paraffin embedded tissue. The purpose of this study was to compare the cell kinetic data obtained by the BrdU labeling study and the PCNA method in the same patient. The relationship between labeling indices of BrdU incorporated into S-phase and PCNA expressed by cycling cells was investigated in 31 patients with brain tumors. Both of the labeling indices showed good correlation with histological grade of the tumor. The values of the PCNA labeling index (LI) were parallel but higher than those of the BrdU LI, and the relation PCNA LI = 2.2 x BrdU LI + 0.8 (r2 = 0.86) was obtained. The results of this study show that PCNA could replace the BrdU method for identifying the proliferating cells, and the major advantages of PCNA method is that it could be done without any pretreatment and avoid injection of the teratogenic agent for diagnostic purpose.

Cell kinetics of human transitional cell carcinoma of the bladder by in vitro bromodeoxyuridine labeling / 대한비뇨기과학회지

We labeled S-phase cell in human bladder tumors using anti-bromodeoxyuridine (BrdU) antibody. The thymidine analogue, BrdU, was incorporated into DNA synthesis phase by in vitro labeling technique. Separate samples of 27 transitional cell bladder cancers were labeled, fixed with 7O per cent ethanol, embedded in paraffin, sectioned and stained by an immunoperoxidase method with anti-bromodeoxyuridine monoclonal antibody as the first antibody. The bromodeoxyuridine labeling index (LI), S phase fraction, was determined by counting the number of bromodeoxyuridine labeled cells in the tissue sections. The average LI in normal and bladder tumors were 2.99% and 11.8% respectively. All grade I and grade II tumors had a LI lower than 10%. while the average LI in grade III tumors was 2l.46%. The average LI for noninvasive (Ta and T1) and invasive tumors were 7.75 % and 26% respectively. The average LI in recurrent tumors were 15.37% compared to 7.76% in initial tumors. The high labeling index in tumors tissue appears to indicate a more malignant potential and thus a poor prognosis. Although further studies are required for its full significance, the measurement of BrdU labeling index by in vitro method may be a quantitative assay of biological potential of individual tumors and has practical value in the management of transitional cell carcinoma of the bladder.

DNA Ploidy and S-Phase Fraction in Proliferative Hepatic Lesions of Rat Liver Induced by Dietylnitrosamine and Partial Hepatectomy

We have investigated the changes of DNA ploidy and S-phase fraction in proliferative lesions of rat liver. Proliferative lesions were induced by diethylnitrosamine and partial hepatectomy. DNA ploidy was measured by flow cytometer, and S-phase fraction was measured by in situ bromodeoxyuridine(BRdU)-anti BRdU monoclonal antibody techniques. Normal liver and initiated lesion revealed DNA diploidy or DNA tetraploidy. Hepatocyte nodule (NODULE) and hepatocelular carcinoma (HCC) revealed DNA diploidy, tetraploidy or aneuploidy. S-phase fraction was 1.0+/-0.9, 1.0+/-0.9m 3.7+/-2.3, 5.5+/-4.9, and 13.8+/-11.6 in normal liver, initiated lesion, NODULE not associated with HCC, NODULE associated with HCC, and HCC, respectively. In NODULE associated with HCC, it was widely distributed, ranging from 0.8 to 15.5%. In conclusion, S-phase fraction appeared to be increased as the hepatocarcinogenesis proceeded, but DNA ploidy did not. There was a heterogeneity of DNA ploidy and S-phase fraction in the proliferative hepatic lesions.