Objective: To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers...Objective: To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers. Methods: Tumor tissue samples of lung cancers and paired non-tumor tissues of the lung were obtaimed from 31 lung cancer patients. Total RNA was extracted and cDNA was synthesized. Nested polymernse chain reaction amplification using MAGE-A3 specific primer was performed to detect the expression of MAGE-A3. The 10 clones of 5 samples of MAGE-A3 mRNA positive PCR products were DNA sequenced by using DNAs sequencer (PE-377). Results: Of 31 lung cancers, 26 (83.9%) expressed MACE-A3 mRNA. The expression of MAGE-A3 gene was not detectable in the adjacent lung tissues. The DNA sequencing confirmed that the target gene fragment in all 5 samples of PCR products was MACE-A3 cDNA. Point nmtations occurred in 4 samples (8 clones) detected (C^2773→T^2773; G^2807→A^2807) resulting in alternation of amino acid residue in one position (E^143→K). Conclusion: (1) The MAGE-A3 gene was expressed exclusively in tumor tissues of the patients with lung cancer in China. This tumor rejection antigen may have potential to be used as a new peptide vaccine for immunotherapy for lung eancers. (2) There are two point mutations of MAGE-A3 gene sequence in some Chinese lung cancer patients.展开更多
AIM: To evaluate whether folate levels in mucosal tissue and some common methylenetetrahydrofolate reductase (MTHFR) variants are associated with the risk of gastric cancer through DNA methylation. METHODS: Real-time ...AIM: To evaluate whether folate levels in mucosal tissue and some common methylenetetrahydrofolate reductase (MTHFR) variants are associated with the risk of gastric cancer through DNA methylation. METHODS: Real-time PCR was used to study the expression of tumor related genes in 76 mucosal tissue samples from 38 patients with gastric cancer. Samples from the gastroscopic biopsy tissues of 34 patients with chronic superficial gastritis (CSG) were used as controls. Folate concentrations in these tissues were detected by the FOL ACS: 180 automated chemiluminescence system. MTHFR polymorphisms were analyzed by PCR-RFLP, and the promoter methylation of tumor-related genes was determined by methylation-specific PCR (MSP). RESULTS: Folate concentrations were significantly higher in CSG than in cancerous tissues. Decreased expression and methylation of c-myc accompanied higher folate concentrations. Promoter hypermethylation and loss of p16INK4A in samples with MTHFR 677CC were more frequent than in samples with the 677TT or 677CT genotype. And the promoter hypermethylation and loss of p21WAF1 in samples with MTHFR 677CT were more frequent than when 677CC or 677TT was present. The 677CT genotype showed a non-significant higher risk for gastric cancer as compared with the 677CC genotype. CONCLUSION: Lower folate levels in gastric mucosal tissue may confer a higher risk of gastric carcinogenesisthrough hypomethylation and overexpression of c-myc.展开更多
AIM:To prepare and identify specific anti-mouse caspase-12 hammerhead ribozymes in vitro, in order to select a more effective ribozyme against mouse caspase-12 as a potential tool to rescue cells from endoplasmic reti...AIM:To prepare and identify specific anti-mouse caspase-12 hammerhead ribozymes in vitro, in order to select a more effective ribozyme against mouse caspase-12 as a potential tool to rescue cells from endoplasmic reticulum stress induced apoptosis. METHODS: Two hammerhead ribozymes directed separately against 138 and 218 site of nucleotide of mouse caspase-12 mRNA were designed by computer software, and their DNA sequences were synthesized. The synthesized ribozymes were cloned into an eukaryotic expression vector-neorpBSKU6 and embedded in U6 SnRNA context for further study. Mouse caspase-12 gene segment was cloned into PGEM-T vector under the control of T7 RNA polymerase promoter (containing gene sequence from positions nt 41 to nt 894) as target. In vitro transcription both the ribozymes and target utilize T7 promoter. The target was labeled with [α-32P]UTP, while ribozymes were not labeled. After gel purification the RNAs were dissolved in RNase free water. Ribozyme and target were incubated for 90 min at 37 ℃ in reaction buffer (40 mmol/L Tris-HCL, pH 7.5, 10 mmol/L Mg2+). Molar ratio of ribozyme vs target was 30:1. Samples were analyzed on 6% PAGE (containing 8 mol/L urea). RESULTS: Both caspase-12 and ribozyme gene sequences were successfully cloned into expression vector confirmed by sequencing. Ribozymes and caspase-12 mRNA were obtained by in vitro transcription. Cleavage experiment showed that in a physiological similar condition (37 ℃, pH 7.5), Rz138 and Rz218 both cleaved targets at predicted sites, for Rz138 the cleavage efficiency was about 100%, for Rz218 the value was 36.66%. CONCLUSION: Rzl38 prepared in vitro can site specific cleave mouse caspase-12 mRNA with an excellent efficiency. It shows a potential to suppress the expression of caspase-12 in vivo, thus provided a new way to protect cells from ER stress induced apoptosis.展开更多
In-situ 2 vol.%TiB2 particle reinforced Al−xSi−0.3Mg(x=7,9,12,15 wt.%)composites were prepared by the salt−metal reaction,and the microstructures and mechanical properties were investigated.The results show that the T...In-situ 2 vol.%TiB2 particle reinforced Al−xSi−0.3Mg(x=7,9,12,15 wt.%)composites were prepared by the salt−metal reaction,and the microstructures and mechanical properties were investigated.The results show that the TiB2 particles with a diameter of 20−80 nm and the eutectic Si with a length of 1−10μm are the main strengthening phases in the TiB2/Al−xSi−0.3Mg composites.The TiB2 particles promote grain refinement and modify the eutectic Si from needle-like to short-rod shape.However,the strengthening effect of TiB2 particles is weakened as the Si content exceeds the eutectic composition,which can be attributed to the formation of large and irregular primary Si.The axial tensile test results and fractography observations indicate that these composites show more brittle fracture characteristics than the corresponding alloy matrixes.展开更多
Objective: Invasion and metastasis are the most significant and intrinsic biological characteristics of cancers, also which are main factors of malignant tumor causing treatment failure and death. Recent studies have...Objective: Invasion and metastasis are the most significant and intrinsic biological characteristics of cancers, also which are main factors of malignant tumor causing treatment failure and death. Recent studies have found that Fra-1 plays an important role on cell migration, invasion, and maintaining malignant phenotype of transformed cells. But there are few stud- ies about the expression and location of Fra-1 in breast tissues and cells being reported .This study just aims to discuss the expression and location of transcription factor Fra-1 in benign and malignant human breast tissues. Methods: The expression of Fra-1 was investigated by immunohistochemistry in neoplastic breast diseases ranging from benign fibroadenoma to very aggressive undifferentiated carcinoma. The correlations of Fra-1 expression with other indicators of breast carcinoma prog- nosis (ER, PR and ErbB2 receptors) were analyzed. Results: All neoplastic breast tissues, either benign or malignant breast tissues, were nuclear immunoreactive for Fra-l-recognizing antibody. In 85% of benign tumors (17/20), the immunoreactive for Fra-l-recognizing antibody as exclusively restricted to the nuclei. In three cases (3/20, 15%), focal unequivocal cytoplas- mic staining was also exhibited. Strong positive nuclear staining for Fra-1 was easily seen in all types of breast carcinomas. However the nucleaflcytoplasmic concomitant immunoreactivity was observed in all types of breast carcinomas. A clear shift in Fra-1 immunoreactivity, from an exclusively nuclear to a simultaneous nuclear and cytoplasmic localization was noticed in 90.2% (37/41) of breast carcinomas. No inverse relationship between Fra-1 and ER and PR protein levels was noticed in malignant tumors. The relative expression level of Fra-1 was not correlated with the expression of ErbB2. Conclusion: The overall expression, pattern and intensity of Fra-1 proteins were correlated with breast oncogenesis. Overexpression of Fra-1, leading to a persistent high cytoplasmic accumulation, may play a role in the process of breast carcinogenesis.展开更多
A novel biomaterial scaffold was created from collagen chitosan/GAG. Its tensile strength was 8.6MPa(wet state)and degree of swelling water was 60%~75% with higer ultimate elongation 300%. Rabbit corneas of collagen ...A novel biomaterial scaffold was created from collagen chitosan/GAG. Its tensile strength was 8.6MPa(wet state)and degree of swelling water was 60%~75% with higer ultimate elongation 300%. Rabbit corneas of collagen chitosan/GAG implantation samples in vivo for biodegradation showed that the inplantion samples was complets biodegrable and digested afere 120 day. There was enought time to maintain cell growth,immigrating and proliferation. This biomaterials scaffold can be used for cell culture and in various tissue engineering fields.展开更多
Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study...Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study investigated the expression of Elf-1 and Ki-67 in non-small-cell lung cancer(NSCLC) and studied their correlation with the clinicopathological features. Methods: Tissue microarray from 64 cases lung cancer tissue and 10 cases normal lung tissue was constructed, immunohistochemical method was used to evaluate the protein expressions of Elf-1 and Ki-67, correlations of the expression of Elf-1 and Ki-67 to clinicopathological features of NSCLC were analyzed. Results: Expression of Elf-1 and Ki-67 in NSCLC tissues were significantly higher than in normal lung tissues(P < 0.05), the positive rate of Elf-1 and Ki-67 was 73.44% and 64.06% in NSCLC group, Overexpression of Elf-1 in NSCLC was significantly related to histopathological grading, different clinical staging and the intensity of ELF-1 expression was significantly higher in the group with lymph node metastasis than that without(P < 0.05). Overexpression of Ki-67 was also closely related to tumor differentiation, clinical stages and lymph node metastasis(P < 0.05). In addition positive correlation was found between the expressive intensity of Elf-1 and Ki-67(τ = 0.295, P = 0.018). Conclusion: The high expression and positive correlation of Elf-1 and Ki-67 in NSCLC suggest that they probably play a role in onset and progression of lung cancer, united detecting their expression could be used as an valuable molecular biological index for predicting the malignant behavior and early diagnosis of NSCLC.展开更多
Objecrive: To investigate the relationship between apoptosis and proliferating cell nuclear antigcn (PCNA)expression of keratinocytes in Condylomata acuminata (CA). Methods: PCNA expression was observed byimmunohistoc...Objecrive: To investigate the relationship between apoptosis and proliferating cell nuclear antigcn (PCNA)expression of keratinocytes in Condylomata acuminata (CA). Methods: PCNA expression was observed byimmunohistochemistry technique (ABC method) in 51 CAspecimens and 1 normal specimens of foreskin or vaginalmucosae. 55 specimens (40 in the CA group and 15 in thecontrol group) were randomly sampled for in situ labelingof apoptotic cells using the TUNEL method. Results: Positive expression of PCNA in CA and controlgroups were 90.2% and 77.8%, respectively, and theproliferation index in CA group was significantly higherthan that in the control group (P<0.001). The positive rateof apoptosis was 42.5% in the LA group and 53.3% in thecontrol group, and there were no significant differences inthe apoptotic index and apoptosis-proliferation ratiobetween two groups (P>0.05). The proliferation indexshowed a significant negativc correlation with theapoptosis-proliferation ratio (r=-0.62, P=0.01) in the CAgrp. Conclusion: It is suggested that the proliferativeappearance of CA could be due to the imbalance betweencell growth and cell death which is caused by moreproliferation and less apoptosis in keratinocytes.展开更多
To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expres...To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. Results: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro α1(Ⅲ) in fibroblasts and some chondrocyte like cells were dominant; and at the end of second week high expression of type Ⅱ procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type Ⅰ procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus.Conclusions: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional technique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells.展开更多
基金This project was supported by the key project of Scientific Committee of Henan Province (No. 0124170232), the key project ofZhengzhou Scientific Committee (No. 04BA60ABYD18), and Tumor Biology Subproject of 211 project of zhengzhou Univevsity
文摘Objective: To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers. Methods: Tumor tissue samples of lung cancers and paired non-tumor tissues of the lung were obtaimed from 31 lung cancer patients. Total RNA was extracted and cDNA was synthesized. Nested polymernse chain reaction amplification using MAGE-A3 specific primer was performed to detect the expression of MAGE-A3. The 10 clones of 5 samples of MAGE-A3 mRNA positive PCR products were DNA sequenced by using DNAs sequencer (PE-377). Results: Of 31 lung cancers, 26 (83.9%) expressed MACE-A3 mRNA. The expression of MAGE-A3 gene was not detectable in the adjacent lung tissues. The DNA sequencing confirmed that the target gene fragment in all 5 samples of PCR products was MACE-A3 cDNA. Point nmtations occurred in 4 samples (8 clones) detected (C^2773→T^2773; G^2807→A^2807) resulting in alternation of amino acid residue in one position (E^143→K). Conclusion: (1) The MAGE-A3 gene was expressed exclusively in tumor tissues of the patients with lung cancer in China. This tumor rejection antigen may have potential to be used as a new peptide vaccine for immunotherapy for lung eancers. (2) There are two point mutations of MAGE-A3 gene sequence in some Chinese lung cancer patients.
基金Supported by the National Basic Research Funds of China 973 Project, No. 2005CB522400 grants from the National Natural Science Foundation of China, No. 30470781 grants from Shanghai Municipal Commission for Science and Technology, No. 04DZ14006 and Doctoral Funds from the Ministry of Education of China, No. 20050266013
文摘AIM: To evaluate whether folate levels in mucosal tissue and some common methylenetetrahydrofolate reductase (MTHFR) variants are associated with the risk of gastric cancer through DNA methylation. METHODS: Real-time PCR was used to study the expression of tumor related genes in 76 mucosal tissue samples from 38 patients with gastric cancer. Samples from the gastroscopic biopsy tissues of 34 patients with chronic superficial gastritis (CSG) were used as controls. Folate concentrations in these tissues were detected by the FOL ACS: 180 automated chemiluminescence system. MTHFR polymorphisms were analyzed by PCR-RFLP, and the promoter methylation of tumor-related genes was determined by methylation-specific PCR (MSP). RESULTS: Folate concentrations were significantly higher in CSG than in cancerous tissues. Decreased expression and methylation of c-myc accompanied higher folate concentrations. Promoter hypermethylation and loss of p16INK4A in samples with MTHFR 677CC were more frequent than in samples with the 677TT or 677CT genotype. And the promoter hypermethylation and loss of p21WAF1 in samples with MTHFR 677CT were more frequent than when 677CC or 677TT was present. The 677CT genotype showed a non-significant higher risk for gastric cancer as compared with the 677CC genotype. CONCLUSION: Lower folate levels in gastric mucosal tissue may confer a higher risk of gastric carcinogenesisthrough hypomethylation and overexpression of c-myc.
基金Supported by the National Natural Science Foundation of China, No. 30170850Shanghai Education Foundation
文摘AIM:To prepare and identify specific anti-mouse caspase-12 hammerhead ribozymes in vitro, in order to select a more effective ribozyme against mouse caspase-12 as a potential tool to rescue cells from endoplasmic reticulum stress induced apoptosis. METHODS: Two hammerhead ribozymes directed separately against 138 and 218 site of nucleotide of mouse caspase-12 mRNA were designed by computer software, and their DNA sequences were synthesized. The synthesized ribozymes were cloned into an eukaryotic expression vector-neorpBSKU6 and embedded in U6 SnRNA context for further study. Mouse caspase-12 gene segment was cloned into PGEM-T vector under the control of T7 RNA polymerase promoter (containing gene sequence from positions nt 41 to nt 894) as target. In vitro transcription both the ribozymes and target utilize T7 promoter. The target was labeled with [α-32P]UTP, while ribozymes were not labeled. After gel purification the RNAs were dissolved in RNase free water. Ribozyme and target were incubated for 90 min at 37 ℃ in reaction buffer (40 mmol/L Tris-HCL, pH 7.5, 10 mmol/L Mg2+). Molar ratio of ribozyme vs target was 30:1. Samples were analyzed on 6% PAGE (containing 8 mol/L urea). RESULTS: Both caspase-12 and ribozyme gene sequences were successfully cloned into expression vector confirmed by sequencing. Ribozymes and caspase-12 mRNA were obtained by in vitro transcription. Cleavage experiment showed that in a physiological similar condition (37 ℃, pH 7.5), Rz138 and Rz218 both cleaved targets at predicted sites, for Rz138 the cleavage efficiency was about 100%, for Rz218 the value was 36.66%. CONCLUSION: Rzl38 prepared in vitro can site specific cleave mouse caspase-12 mRNA with an excellent efficiency. It shows a potential to suppress the expression of caspase-12 in vivo, thus provided a new way to protect cells from ER stress induced apoptosis.
基金The authors are grateful for the financial supports from the National Natural Science Foundation of China(51804349)the China Postdoctoral Science Foundation(2018M632986)the Natural Science Foundation of Hunan Province,China(2019JJ50766).
文摘In-situ 2 vol.%TiB2 particle reinforced Al−xSi−0.3Mg(x=7,9,12,15 wt.%)composites were prepared by the salt−metal reaction,and the microstructures and mechanical properties were investigated.The results show that the TiB2 particles with a diameter of 20−80 nm and the eutectic Si with a length of 1−10μm are the main strengthening phases in the TiB2/Al−xSi−0.3Mg composites.The TiB2 particles promote grain refinement and modify the eutectic Si from needle-like to short-rod shape.However,the strengthening effect of TiB2 particles is weakened as the Si content exceeds the eutectic composition,which can be attributed to the formation of large and irregular primary Si.The axial tensile test results and fractography observations indicate that these composites show more brittle fracture characteristics than the corresponding alloy matrixes.
文摘Objective: Invasion and metastasis are the most significant and intrinsic biological characteristics of cancers, also which are main factors of malignant tumor causing treatment failure and death. Recent studies have found that Fra-1 plays an important role on cell migration, invasion, and maintaining malignant phenotype of transformed cells. But there are few stud- ies about the expression and location of Fra-1 in breast tissues and cells being reported .This study just aims to discuss the expression and location of transcription factor Fra-1 in benign and malignant human breast tissues. Methods: The expression of Fra-1 was investigated by immunohistochemistry in neoplastic breast diseases ranging from benign fibroadenoma to very aggressive undifferentiated carcinoma. The correlations of Fra-1 expression with other indicators of breast carcinoma prog- nosis (ER, PR and ErbB2 receptors) were analyzed. Results: All neoplastic breast tissues, either benign or malignant breast tissues, were nuclear immunoreactive for Fra-l-recognizing antibody. In 85% of benign tumors (17/20), the immunoreactive for Fra-l-recognizing antibody as exclusively restricted to the nuclei. In three cases (3/20, 15%), focal unequivocal cytoplas- mic staining was also exhibited. Strong positive nuclear staining for Fra-1 was easily seen in all types of breast carcinomas. However the nucleaflcytoplasmic concomitant immunoreactivity was observed in all types of breast carcinomas. A clear shift in Fra-1 immunoreactivity, from an exclusively nuclear to a simultaneous nuclear and cytoplasmic localization was noticed in 90.2% (37/41) of breast carcinomas. No inverse relationship between Fra-1 and ER and PR protein levels was noticed in malignant tumors. The relative expression level of Fra-1 was not correlated with the expression of ErbB2. Conclusion: The overall expression, pattern and intensity of Fra-1 proteins were correlated with breast oncogenesis. Overexpression of Fra-1, leading to a persistent high cytoplasmic accumulation, may play a role in the process of breast carcinogenesis.
文摘A novel biomaterial scaffold was created from collagen chitosan/GAG. Its tensile strength was 8.6MPa(wet state)and degree of swelling water was 60%~75% with higer ultimate elongation 300%. Rabbit corneas of collagen chitosan/GAG implantation samples in vivo for biodegradation showed that the inplantion samples was complets biodegrable and digested afere 120 day. There was enought time to maintain cell growth,immigrating and proliferation. This biomaterials scaffold can be used for cell culture and in various tissue engineering fields.
基金Supported by grants from the Science and Technology Development Project of Yantai(No.2008162)Natural Sciences Foundation of Shandong(No.ZR2013HL004)Science and Technology Important Project of Binzhou Medical College(No.BY20bkjzd04)
文摘Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study investigated the expression of Elf-1 and Ki-67 in non-small-cell lung cancer(NSCLC) and studied their correlation with the clinicopathological features. Methods: Tissue microarray from 64 cases lung cancer tissue and 10 cases normal lung tissue was constructed, immunohistochemical method was used to evaluate the protein expressions of Elf-1 and Ki-67, correlations of the expression of Elf-1 and Ki-67 to clinicopathological features of NSCLC were analyzed. Results: Expression of Elf-1 and Ki-67 in NSCLC tissues were significantly higher than in normal lung tissues(P < 0.05), the positive rate of Elf-1 and Ki-67 was 73.44% and 64.06% in NSCLC group, Overexpression of Elf-1 in NSCLC was significantly related to histopathological grading, different clinical staging and the intensity of ELF-1 expression was significantly higher in the group with lymph node metastasis than that without(P < 0.05). Overexpression of Ki-67 was also closely related to tumor differentiation, clinical stages and lymph node metastasis(P < 0.05). In addition positive correlation was found between the expressive intensity of Elf-1 and Ki-67(τ = 0.295, P = 0.018). Conclusion: The high expression and positive correlation of Elf-1 and Ki-67 in NSCLC suggest that they probably play a role in onset and progression of lung cancer, united detecting their expression could be used as an valuable molecular biological index for predicting the malignant behavior and early diagnosis of NSCLC.
文摘Objecrive: To investigate the relationship between apoptosis and proliferating cell nuclear antigcn (PCNA)expression of keratinocytes in Condylomata acuminata (CA). Methods: PCNA expression was observed byimmunohistochemistry technique (ABC method) in 51 CAspecimens and 1 normal specimens of foreskin or vaginalmucosae. 55 specimens (40 in the CA group and 15 in thecontrol group) were randomly sampled for in situ labelingof apoptotic cells using the TUNEL method. Results: Positive expression of PCNA in CA and controlgroups were 90.2% and 77.8%, respectively, and theproliferation index in CA group was significantly higherthan that in the control group (P<0.001). The positive rateof apoptosis was 42.5% in the LA group and 53.3% in thecontrol group, and there were no significant differences inthe apoptotic index and apoptosis-proliferation ratiobetween two groups (P>0.05). The proliferation indexshowed a significant negativc correlation with theapoptosis-proliferation ratio (r=-0.62, P=0.01) in the CAgrp. Conclusion: It is suggested that the proliferativeappearance of CA could be due to the imbalance betweencell growth and cell death which is caused by moreproliferation and less apoptosis in keratinocytes.
文摘To analyze the expression of procollagen gene in fracture callus, and to search for the technique of in situ hybridization for undecalcified skeletal tissue. Methods: In situ hybridization of procollagen gene expression was performed on the undecalcified cryosections of rat fracture callus at 7, 14, and 28 d. Results: The hybridization signals achieved were clear and easy to be localized with high specificity. On the 7th day, the expressions of pro α1(Ⅲ) in fibroblasts and some chondrocyte like cells were dominant; and at the end of second week high expression of type Ⅱ procollagen mRNA was observed in chondrocytes. At the end of fourth week, the cartilaginous callus was almost all replaced by woven bone tissue, and some type Ⅰ procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus.Conclusions: The modified method employed in this study is easier, quicker, and more sensitive with high specificity than the conventional technique for in situ hybridization of procollagen gene expression of decalcified rat fracture callus. The phenomenon of shared phenotype expression, which was demonstrated among cells engaged in fracture healing, indicates an important approach to reveal the mechanism of the origin, differentiation, and orientation of cells.