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2009, Volume 40 Issue 6
06 December 2009 Previous Issue    Next Issue
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P>Stereological study of the age-related changes of the myelinated nerve fibers in the hippocampal formation of rat /P> 2009, 40 (6):  851-856.  doi: 10.3969/j.issn.0529-1356.2009.06.001 Abstract ( )   Objective To explore the changes of the hippocampal formation and the myelinated nerve fibers in the hippocampal formation of aged female Long-Evans rat. Methods The hippocampal formation and the myelinated nerve fibers in the hippocampal formation of 5 young (6-month old), 5 middle-aged (18-month old) and 6 aged (28-month old) female Long-Evans rats were quantitatively investigated with the stereological techniques and transmission electronic microscope technique. Results There were no significant changes in the volume of hippocampal formation, the volume density, the total volume, the length density and the mean diameter of the myelinated nerve fibers in the hippocampal formation among young, middle-aged and aged rats. When compared to that of young rats, the total length of the myelinated nerve fibers in the hippocampal formation of middle-aged rats was significantly increased by 63.6%. When compared to that of middle-aged rats, the total length of the myelinated nerve fibers in the hippocampal formation of aged rats was significantly decreased by 47.5%. When compared to that of young rats, the total length of the myelinated nerve fibers in the hippocampal formation of aged rats was non-significantly decreased by 13.8%. Conclusion Although the reason why the total length of the myelinated nerve fibers in the hippocampal formation of middle-aged rats was longer than that of young rats needs further investigations, the present results together with our previous findings in white matter and cortex further suggest that there are age-related changes of the myelinated nerve fibers in the normal aged brains. Related Articles | Metrics

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Expression of phosphoryl extracellular signalregulated kinases in the differentiation of NSCs into neurons induced by the extracts of the fimbria-transected hippocampi in rat 2009, 40 (6):  857-861.  doi: 10.3969/j.issn.0529-1356.2009.06.002 Abstract ( )   Objective To investigate the effect of extracellular signal-regulated kinases(ERK) signal transduction in the process of NSCs differentiating into neurons in the fimbria-transected hippocampi’s extracts. Methods Twelve Sprague-Dawley rats’right fimbrias were transected. The extracts were gained from the fimbria-transected hippocampi at the 14th day normal rat, and the extracts supernatant fluid was collected after centrifugal process, then the protein concentration in the extracts was determined. In the serum-free medium，NSCs from the fetal hippocampus were planted on 24 well culture plate, then were divided into three group and eight wells for each group as follows: the transected group contained the extracts of the fimbria-transected hippocampi; the normal group contained the extracts of the normal hippocampi; the pure control group have no extracts. After cultured for 14 days,the cells were detected by using MAP-2 and p-ERK immunofluorescence. Result The number, area, perimeter of MAP-2 positive neurons were all declined in transected group, the normal group and the control group orderly. Statistic results showed significant difference between every two groups. The number of MAP-2/p-ERK double-positive neurons were decreased in transected group, the normal group and the control group orderly, but the percentage of double-labeled neurons in total MAP-2 positive neurons were increased in turn. In these two aspect, there were also significant difference between every two group. And most of the MAP-2/p-ERK double-positive neurons were immature. Conclusion The extracts of the fimbria-transected hippocampi had obvious effects on promoting NSCs differentiating into neurons and speeding up the maturation of neurons than those of the normal hippocampi. The morphological results showed that ERK signal transduction might be related to the differentiation of NSCs i Related Articles | Metrics

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DIV>Effects of heroin and ephedrine on the histological structure and ChAT activity of hypothalamus and hippocampus of filial mice /DIV> 2009, 40 (6):  862-868.  doi: 10.3969/j.issn.0529-1356.2009.06.003 Abstract ( )   Objective To explore effects of heroin and ephedrine on the histological structure and ChAT activity of hypothalamus and hippocampus of filial mice. Expression of Bcl-2 associated X protein(Bax protein) and keratinocyte growth factor(KGF) of hypothalamus and hippocampus were measured. Methods One hundred and eight filial mice were given intraperitoneal injection of heroin and ephedrine by gradually increase of doses, apoptosis and expression of Bax protein and KGF of hypothalamus and hippocampus were observed by Giemsa staining and immunohistochemistry, and the ChAT activity was detected by colorimetry. Results After administration of heroin and ephedrine at 5,10,15,20 days, the number of apoptotic cells and expression of Bax protein and KGF of hypothalamus and hippocampus were significantly increased and ChAT activity was lower than those of the control group(EM>P/EM><0.05 or EM>P/EM><0.01). There were differences between heroin group and the ephedrine group in the abovementioned four indexes (EM>P/EM><0.05 or EM>P/EM><0.01). The number of apoptotic cells and Bax protein and KGF immunopositive neurons of hypothalamus and hippocampus increased by the increase in dose of heroin and ephedrine. ConclusionsHeroin and ephedrine had great effect on the histological structure and ChAT activity of hypothalamus and hippocampus of filial mice, and this effects would be related to the cell apoptosis of hypothalamus and hippocampus. Related Articles | Metrics

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Expression and significance of cytochrome C and Bax in hippocampus of posttraumatic stress disorder rat 2009, 40 (6):  869-872.  doi: 10.3969/j.issn.0529-1356.2009.06.004 Abstract ( )   Objective To observe the expression of cytochrome C （Cyt C）and apoptosis-related gene Bax in hippocampus of posttraumatic stress disorder（PTSD）rats in order to investigate their effects and relationships. Methods Sixty male Wistar rats were used in the present study. The single-prolonged stress（SPS）-method was carried out to set up the rat PTSD models. There were six groups：after SPS 1，4，7，14，28 days groups and control group. The expression of Cyt C and Bax proteins were detected with immunohistochemistry, double immunofluorescence, confocal laser scanning microscopy and Western blotting. Results The expression of Cyt C peaked at 4 days and maintained higher level at 7 days after SPS. At 7 days after SPS, strong immunoreactivity of Bax was noticed,At 14 days after SPS, the expression of Bax was down-regulated and then gradually decreased. Conclusion Bax is Related Articles | Metrics

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Changes of nuclear receptors-glucocorticoid receptors in locus ceruleus neurons of posttraumatic stress disorder-like rat 2009, 40 (6):  873-876.  doi: 10.3969/j.issn.0529-1356.2009.06.005 Abstract ( )   Objective To investigate expression changes of nuclear receptors-glucocorticoid receptors (GR) in the locus ceruleus neurons of posttraumatic stress disorder(PTSD)like rats. Methods Sixty adult healthy male Wistar rats were enrolled in the experiment. PTSD-like rat model was established by single prolonged stress (SPS). There was no SPS stimulation in the control group and the rats of the other five groups were undisturbedly raised for 24 hours, 4 days,7 days,14 days and 28 days respectively after SPS stimulation.The expressions of glucocorticoid receptors in the locus ceruleus nucleus of all groups were detected with Nissl staining, immunohistochemistry,Western blotting and PCR, and image analysis and statistical analysis were performed. Results GR was distributed in the nucleus of coeruleus neurons, GR expression was showed after 24 hours, 4 days, 7 days treatment and gradually increased, restorative downregulation was seen after 14 days and 28 days, but still high(EM>P/EM><0.05). Conclusion After SPS, the GR locus ceruleus temporarily increased and then lowered, suggesting that PTSD rat locus coeruleus neurons in nuclear receptor-GR expression changes may directly involve in the continuing spirit of PTSD symptoms occurred in the development process Related Articles | Metrics

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Expressions of CaSUP>2+/SUP>/calmodulin in hippocampus of rats with posttraumic stress disorder 2009, 40 (6):  877-880.  doi: 10.3969/j.issn.0529-1356.2009.06.006 Abstract ( )   Objective To observe the changes of intracellular free calcium and the expression of CaM in the hippocampal neurons of posttraumatic stress disorder (PTSD) rats and to further investigate the neurobiological mechanisms. Methods The SPS-method was used to set up the rat PTSD models. A total of sixty male Wistar rats were randomly divided into 12 hours,1 day,4 days,7 days groups of SPS and normal control group. The intracellular free calcium was examined by fluorescence spectrophotometer. The expression of CaM was detected by using immunohistochemistry, Western blotting and RT-PCR. Results The intracellular free calcium level in the hippocampus of experimental rats was markedly increased 12 hours after SPS stimulation,and reached the peak after 1 day, then gradually decreased to normal level after 7 days. The expression of CaM in the hippocampus 1day after SPS was also the highest and then gradually decreased.Conclusion The lasting dysfunction of CaSUP>2+/SUP>CaM signaling cascades in hippocampal may play important roles in the pathogenesis of PTSD rats. Related Articles | Metrics

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Changes of behavior, CaSUP>2+/SUP>/CaMKⅡ in hippocampus of rats with chronic forced swimming stress model 2009, 40 (6):  881-885.  doi: 10.3969/j.issn.0529-1356.2009.06.007 Abstract ( )   Objective To observe the changes of behavior, intracellular free calcium and the expression of calmodulin dependent protein kinase Ⅱ(CaMKⅡ) in the hippocampal neurons of chronic forced swimming stress rats. Methods Male Wistar rats were randomly divided into control group and chronic forced swimming stress group. The behavior was examined using sucrose preference test, open-filed test and Morris water maze. The intracellular free calcium was examined by fluorescence spectrophotometer. The expression of CaMKⅡ was detected using colloidal gold immunoelectron microscopy technique, Western blotting and RT-PCR. Results The consumption of sucrose and erect quantity of chronic forced swimming stress group were lower than those of control group（EM>P/EM><0.01, EM>P/EM><0.05）. The escape latency time in Morries water maze test of chronic forced swimming stress group was higher than that of control group（EM>P/EM><0.01）. The intracellular free calcium level and the expression of CaMKⅡ in the hippocampus was higher than that of control group（EM>P/EM><0.01）.Conclusion The lasting dys Related Articles | Metrics

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Study of the expression of cyclooxygenase-2, malonic dialdehyde and the protective effect of Tetramethylpyrazine after cerebral ischemic-reperfusion in rat 2009, 40 (6):  886-890.  doi: 10.3969/j.issn.0529-1356.2009.06.008 Abstract ( )   Objective To investigate the expression, relationship, significance of cyclooxygenase-2 (COX-2) and malonic dialdehyde (MDA) after cerebral ischemic-reperfusion injury and provide basis of treatment. Methods The focal cerebral ischemic-reperfusion model was established with thread embolish of middle cerebral artery. Western blotting, barhituric acid method and neurological evaluation were used to examine the expression of COX-2, MDA in cortex and the changes of neurological function； TTC staining was used to observe the changes of cerebral infarction volume. Results COX-2 prorein expression was correclated well with the MDA（EM>r/EM>=0.910，EM>P/EM>0.01). The content of COX-2 and MDA was very low in sham operation group, they were increased significantly at I2h/R6h model group, with the increase of reperfusion time,they remarkably reached its peak at I2h/R24h,they were slightly lower at I2h/R48h, but still maintained at a high level;Compared with model group, in tetramethylpyrazine(TMP) treatment group, the content of MDA and cerebral infarction volume were markedly decreased（EM>P/EM>0.01). Conclusion The expression of COX-2 and MDA increases in cerebral ischemic-reperfusion injury.It indicates they may play an important role in the mechani Related Articles | Metrics

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Construction of a lentivirus vector for RNA interference of myosin Va and its effect on motility and migration of PG cells 2009, 40 (6):  891-896.  doi: 10.3969/j.issn.0529-1356.2009.06.009 Abstract ( )   Objective To construct a lentivirus vector for RNA interference targeting myosin Va gene and to observe its effect on motility and migration of human pulmonary giant cell carcinoma PG cells. Methods Based on the efficient target sequence for myosin Va RNAi, two pairs of oligo DNA containing myosin Va RNAi target sequence or scramble sequence were synthesized and inserted into pSuper vector, followed by sequence analysis. The expressing cassette H1 promoter-shM5A/shCON from the recombinant pSuper plasmid was then transferred to the lentivirus vector plenti4, and the recombinant lentivirus was packaged. PG cells were transduced with the packaged lentivirus and the positive cells were screened by zeocin selection. RT-PCR was performed to determine the myosin Va RNAi efficiency in zeocin-resistant PG cells, and wounding assay and Boyden chamber assay were utilized to examine the capabilities of motility and migration in myosin Va RNAi PG cells. Results Restriction enzyme digestion and sequencing confirmed the successful construction of the lentivirus vector containing myosin Va RNAi target or scramble sequence. RT-PCR result showed that myosin Va mRNA levels were remarkably reduced in lentivirus-based myosin Va RNAi PG cells. The abilities of motility and migration were also significantly inhibited in lentivirus-based myosin Va RNAi PG cells, as demonstrated in wounding assay and Boyden chamber assay.Conclusion Myosin Va RNAi lentivirus vector was successfully constructed and efficiently repressed myosin Va expression in PG cells. Repression of myosin Va by RNAi led to the inhibition of PG cells motility and migration, indicating that there might exist correlation between the expression of myosin Va and cancer progression. Related Articles | Metrics

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Study on the erythroid differentiation on K562 cells induced by angelica polysaccharides and its signal transdution pathway 2009, 40 (6):  897-901.  doi: 10.3969/j.issn.0529-1356.2009.06.010 Abstract ( )   Objective To explore the relative signal transduction pathway of angelica polysaccharide(APS) inducing K562 cells toward erythroid differentiation. Methods K562 cells were divided into control group and ASP group, the control group cells were routinely cultured and APS group cells were treated with APS(final concentration is 100500mg/L). By using of benzidine staining and spectrophotometry, the characteristics of erythroid differentiation of K562 cell induced by APS were detected; By using of laser confocal microscopy, the distribution of JAK-2 and STAT-5 in K562 cells was observed; By using of Western blotting, the expression of JAK-2 and STAT-5 in nucleus and cytoplasm of K562 cells was detected, By using of imm~uno~pre~cipi~ta~tion, the phosphorylation change of JAK-2 in cytoplasm was tested. Results After being induced by APS, the benzidine staining positive rate of K562 was increased.With increasing the concentration of APS, hemoglobin synthesis in K562 cells was promoted accordingly. After being cultured for 12 hours, 24 hours and 48 hour, the expression of STAT-5 in nucleus of K562 cells induced by APS was significantly higher than that of control group, however, expression of STAT-5 in cytoplasm of K562 cell induced by APS was obviously lower. The expression of JAK-2 in K562 cells was not different between APS group and control group, but the JAK-2 phosphorylation level of APS group was much higher than that of control group.Conclusion APS can induce erythroid differentiation of K562 cells, and the mechanisms may be that APS can promote the phosphorylationthe of J Related Articles | Metrics

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Expression of PCNA, lymphatic vessel density, lymphatic vessel area and their relationship with lymphaticmetastasis in the hamster tongue cancer development 2009, 40 (6):  902-908.  doi: 10.3969/j.issn.0529-1356.2009.06.011 Abstract ( )   ［Abstract］ Objective To investigate the relationship of the lymphangiogenesis and lymphatic metastasis by observing the expression of proliferating cell nuclear antigen in the hamster tongue cancer development and analyzing the correlations of lymph vessel density (LVD), lymphatic vessel area (LVA) and malignance or lymphatic metastasis of tongue cancer cells. Methods Forty-two hamster tongue cancer models were induced by painting 1.5% 7,12-dimethylben(a) anthracene (DMBA) acetone solution after scratching, 7 models were sacrificed every two weeks. The pathological changes of tongue cancer were determined by the HE staining. The lymphatic hyperplasia was determined by PCNA and Desmoplakin(DP I +Ⅱ) Immunohistochemical double staining. The tumors and lymphatic endothelial cell proliferation index (proliferation index, PI), lymphatic vessel density (LVD) and lymphatic vessel area (LVA) were measured and statistically analyzed. Results The oncogenesis of tongue was divided into three stages: atypical hyperplasia, carcinoma EM>n situ/EM> and early invasive stage; atypical hyperplasia group：PI (91.55), LVA (7570.23), LVD (2.50); carcinomaEM>in situ/EM> group: PI (113.36), LVA (12105.45), LVD (3.73); early invasive group: PI (124.67), LVA (14524.33), LVD (5.33). The PI, LVA and LVD were compared among groups above (EM>P/EM><0.05), and the correlation of PI with LVA and LVD was positive (EM>P/EM><0.05); Lymphatic metastasis group：PI (130.50), LVA (15430.67), LVD (6.17), non lymphatic metastasis group：PI (113), LVA (12711.67), LVD (3.67), the PI, LVA and LVD were compared between the two groups above (P<0.05).Conclusion With the development of hamster tongue cancer, lymphatic endothelial cell proliferation activity increased, suggesting that the lymphangiogenesis existed in the tongue carcinogenesis process; the values of the LVD and LVA increased in the hamster tongue cancer Related Articles | Metrics

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Effect of fibrinogen gel and chitin on BMSCs to chondrocytes differentiation 2009, 40 (6):  909-913.  doi: 10.3969/j.issn.0529-1356.2009.06.012 Abstract ( )   Objective We compare the effects of fibrinogen gel and chitin on BMSCs to chondrocytes differentiation in order to explore the relationship between threedimensional scaffold and cartilage tissue engineering seed cells BMSCs differentiation. Methods BMSCs together with chitin and fibrin gel complexes were cultured EM>in vitro/EM> and implanted into rat’s articular cartilage defect location. After 14 days EM>in vitro/EM> culture, HE staining, toluidine blue staining and type II collagen immunohistochemical staining were performed; After transplantation EM>in vivo/EM>, for 2 weeks, 4 weeks and 6 weeks, morphology observations, expression of cartilagespecific protein analysis and BMSCs EM>in vivo/EM> tracer method were performed. We analyzed the differentiation of BMSCs into chondrocytes by statistical methods. Results The comparison of positive cell rate of type II collagen immunohistochemical staining in BMSCs-fibrin gel group and BMSCs-chitin group which were cultured EM>in vitro/EM>, showed no significant difference with control group. Integrated absorbance(IEM>A/EM>) change rate of toluidine blue staining and type II collagen immunohistochemical staining in BMSCsfibrin gel group which was cultured EM>in vivo/EM>, showed significantly different with other groups and control group.Conclusion The results showed that EM>in vitro/EM> fibrin gel or chitin has very weak induction of BMSCs to cartilage Related Articles | Metrics

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Inhibition effects of nuclear factor-κB on cytokines releasing from human corneal fibroblasts 2009, 40 (6):  914-918.  doi: 10.3969/j.issn.0529-1356.2009.06.013 Abstract ( )   Objective To investigate the inhibition effects of nuclear factor-κB(NF-κB) on cytokine expression of cultured human corneal fibroblasts in vitro. Methods The cultured human corneal fibroblasts were used in the present experiment. Cell viability insensitive to emodin on cultured human corneal fibroblasts in vitro was assessed using the MTT assay.Cells were randomly divided into three groups：control group（group 0 hour，EM>n/EM>=6）,lipopolysaccharidel (LPS)group （EM>n/EM>=24）and emodin pretreatment group（EM>n/EM>=24）. Cells of the emodin pretreatment group were incubated with emodin for 30 minutes before LPS challenged. The cultured human corneal fibroblasts were then challenged with LPS, the expression of interleukin-8(IL-8) mRNAs was detected by reverse transcription polymerase chain reaction analysis (RT-PCR) and the proteins of IL-8 secreted from these cells were assessed by enzymelinked immunosorbent assays (ELISA). Western blotting analysis was used for detecting the protein expression of inhibitor of nuclear factor-κB-α（IκB-α）induced by LPS.At the same time，the effects of emodin on these expressions were also assessed in fibroblasts. Results The concentration of emodin used in this study was safe to these cultured human corneal fibroblasts in vitro.Both the protein release and mRNA expression of IL-8 in corneal fibroblasts increased significantly after challenged with LPS, however, the protein level of IκB-α significantly decreased after treated with LPS. These results indicated that LPS could mediate the activation of NF-κB and upregulate the expression of cytokines in corneal fibroblasts. Pretreated with emodin 30 min before challenged with LPS, the activation of NF-κB induced by LPS was markedly inhibited, and the Related Articles | Metrics

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ERK signal is pathway involved in mechanical stretch induced HMGB1 expression in alveolar epithelial cells 2009, 40 (6):  919-922.  doi: 10.3969/j.issn.0529-1356.2009.06.014 Abstract ( )   Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10SUP>5 /SUP>cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (EM>P/EM><0.05) and decreased in group C (EM>P/EM><0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (EM>P/EM><0.05) and decreased in group C (EM>P/EM><0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (EM>P/EM><0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway. Related Articles | Metrics

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Fetal rat liver filtrate induces the differentiation of rat bone marrowderived mesenchymal stem cells into hepatocytes 2009, 40 (6):  923-927.  doi: 10.3969/j.issn.0529-1356.2009.06.015 Abstract ( )   Objective To explore the possibility that rat bone mesenchymal cells (BMSCs) can differentiate into hepatocytes under the affection of fetal liver filtrate. Methods PAS and green indigo dye were used to detect glycogen and differential level of hepatocytes, respectively. The concentration of ALT, AST, ALP in the culture supernatant were served as markers of hepaocyte function. Results Fourteen days after induced by the fetal liver filtrate, BMSCs changed their shapes into polygon, oval or round. Some of BMSCs were positive for AFP and ALB at 7 days after induction, then the number of positive cells increased, and most of BMSCs expressed AFP and ALB till 21days. The PAS reaction and indocyanine green(ICG) intaking also appeared at 7days. Enzyme in supernatant such as ALT, AST, ALP were fristly detected at 7days and peaked at 14days，then the level declined. Conclusion The fetal rat liver filtrate was able to induce BMSCs into cells with function and characteristics of hepatocytes. Related Articles | Metrics

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Effect of Grp78 on the activation and expression of ERK 1/2 in human hepatocellular carcinoma tissues 2009, 40 (6):  928-932.  doi: 10.3969/j.issn.0529-1356.2009.06.016 Abstract ( )   Objective We examined the Grp78, ERK1/2 and phospho-ERK1/2 expressions in hepatocellular carcinoma(HCC) tissue samples EM>in vitro/EM>, we interfered the expression of Grp78 in SMMC-7721 cells to explore whether Grp78 is involved in ERK1/2 signal pathway. Methods The Grp78, ERK1/2 and phospho-ERK1/2 expressions were detected by immunohistochemistry and confirmed by Western blotting in 47 HCC tissue samples. The Grp78 expression in SMMC-7721 cells was interfered by plasmid transfection and siRNA, ERK1/2 phosphorylation and expression were determined by Western blotting. Results The Grp78 expression was significantly correlated with ERK1/2 and phospho-ERK1/2 in HCC tissue samples. Overexpression of Grp78 promoted ERK1/2 phosphorylation in SMMC-7721 cells and the increased ERK1/2 phosphorylation was inhibited by Grp78 knockdown. Conclusion Grp78 is involved in the regulation of ERK1/2 signal pathway and might be a potential target for the comprehensive therapy of HCC. Related Articles | Metrics

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Expressions of hepatocyte nuclear factor -1α and hepatocyte nuclear factor -4α in human hepatocellular carcinoma 2009, 40 (6):  933-937.  doi: 10.3969/j.issn.0529-1356.2009.06.017 Abstract ( )   Objective To investigate the expression of hepatocyte nuclear factor-1α (HNF-1α) and hepatocyte nuclear factor-4α (HNF-4α) in human hepatocellular carcinoma (HCC) and explore the function of HNF-1α and HNF-4α during HCC carcinogenesis and development. Methods Twenty-six specimens of hepatocellular carcinoma were collected. The expressions of HNF-1α and HNF-4α in HCC tissues and adjacent non-cancerous tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry staining. Results The mRNA levels of HNF-1α and HNF-4α were significantly lower in HCC tissues than that in adjacent non-cancerous tissues (0.818±0.371 vs. 0.383±0.102 for HNF-1α, P0.05; 0.846±0.384 vs. 0.397±0.105 for HNF-4α, P0.05）.The positive rates of HNF-1α and HNF-4α protein were significantly lower in HCC tissues than in adjacent non-c Related Articles | Metrics

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Effects of Aurora A silence by RNA interference on the apoptosis and proliferation of glioma cells 2009, 40 (6):  938-942.  doi: 10.3969/j.issn.0529-1356.2009.06.018 Abstract ( )   Objective To investigate the inhibitory effect of RNA interference on the expression of Aurora A in U251 cells, and the influence on proliferation and apoptosis of U251 cells. Methods The siRNA specific for Aurora A was synthesized and transfected into U251 cells EM>in vitro/EM>. Aurora A mRNA expression and protein content were detected by RT-PCR and Western blotting respectively. The cell proliferation and apoptosis were observed by methyl thiazolyl tetrazolium(MTT) and flow cytometry(FCM). Transmission electron microscope was used to observe the ultrastructural changes of U251 cells. Results After transfection, the expression level of Aurora A mRNA was significantly decreased(EM>P/EM><0.01), and the protein content of Aurora A was also obviously reduced. The inhibitory rate of cell proliferation reached up to 67.57% 72 hours after transfection, which was significantly higer than that of normal control group(EM>P/EM><0.01). The apoptosis rate of U251 cells was significantly increased from (3.69±0.87)% to (15.34±2.16)% (P<0.01). Under the transmission electron microscope, it was observed that the U251 cells showed typical morphologic changes of apoptosis after transfection, such as karyopyknosis, chromatin condensation and margination, intracytoplasmic vacuoles formed, and apoptotic bodies formed. Conclusion The expression of Aurora A gene can be inhibited by siRNA successfully, and it results in the suppression of cell growth and induce apoptosis of human glioma cells EM>in vitro/EM>. Aurora A may become a new target fo Related Articles | Metrics

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Transgenic mouse fetus generated from embryonic stem cells by tetraploid embryo complementation 2009, 40 (6):  943-948.  doi: 10.3969/j.issn.0529-1356.2009.06.019 Abstract ( )   Objective To use tetraploid embryo complementation combined with gene transfer to produce genetically modified embryonic stem cells (EsCs) clones. Methods In this study, EGFP was introduced into ESCs by electroporation, and transfected positive cells were selected by G418 resistance. The tetraploid embryos were obtained from diploid blastomere electrofusion which preformed at 2-cell stage. Afterwards, 19-21 EGFP-ESCs were inserted into each tetraploid blastocyst cavity by piezo drilled microinjection,then the injected blastocysts were transferred into the uterus of pseudo-pregnancy at 2.5-day or the oviduct of 0.5-day female mice. Results The transfected ESCs maintained normal karyotype even after long-term passage (2EM>n/EM>=40). The rate of fusion was 95.07%, and the developmental rate of tetraploid blastocyst was 95%.Totally 410 injected blastocysts were obtained. Unfortunately, we have not got any vital offsprings, except 151 implantation sites (pseudo-pregnancy 2.5 days:29.41%;the oviduct of half one day:64.37%). Furthermore, scattered EGFP expressions in transgenic fetus were observed under invert fluorescent microscope. Conclusion The transfected ESCs were observed in transgenic fetus, and the implantation rate in oviduct was higher than that in uterine. Related Articles | Metrics

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Pathological changes of diabetic rat thyroid ultrastructure and intervention effects of insulin and aminoguanidine 2009, 40 (6):  949-953.  doi: 10.3969/j.issn.0529-1356.2009.06.020 Abstract ( )   Objective To investigate the thyroid ultrastructural pathological changes of diabetes mellitus (DM) rats as well as the intervention effects of insulin and aminoguanidine. Methods Totally 87 rats were treated with streptozotocin to establish DM animal models and divided into DM group（EM>n/EM>=27）,insulin intervention group(EM>n/EM>=32) and aminoguanidine intervention group(EM>n/EM>=28),25 rats were taken as normal controls. Twelve and 20 weeks after the animal model establishment, animals were sacrificed, thyroid tissue was taken and ultrastructure was observed. Results In the thyroid of DM rats, follicular epithelial cells present as applanate shape, microvilli were depleted, rough endoplasmic reticulum dilated to irregular vesicular. None pinocytotic vacuole and casual primary or secondary lysosome were seen. Follicular cavity was dilated, colloid in the cavity had higher electronic-density. Interstitial edema, capillary base lamian was thickened at different stage. Proteo-substance deposition with granulo-shape, cloud shape or homogeneity appeared. The number of thyroid parafollicular cells increased. But endocrine granule in parafollicular cells was few. When compared with DM group, the thyroid tissue injury of insulin intervention group and aminoguanidine intervention group were lessened to different degree. Conclusion The hypofunctional thyroid follicular cells, large quantity of proteo-substance deposition in the interstitium and increased parafollicular cells of DM rats may be related with hyperglycemia toxicity. Insulin and aminoguanidine treatment have some protection effects. Related Articles | Metrics

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Expression of transforming growth factor-β receptors in the outflow tract of mouse embryonic heart 2009, 40 (6):  954-957.  doi: 10.3969/j.issn.0529-1356.2009.06.021 Abstract ( )   Objective To investigate the spatiotemporal expression patterns of transforming growth factor-β(TGF-β) receptors type Ⅰ and type Ⅱ and their relationships with development of outflow tract(OFT) in mouse embryonic heart. Methods Serial sections of mouse embryos from embryonic day 9 (E9d) to embryonic day 14 (E14d) were stained using PAP immunohistochemical methods. Results Expressions of TGF-β receptors type Ⅰ (TGF-βRⅠ) and type II(TGF-βRⅡ) in the myocardial wall of OFT started at E10d, reached the reflection with splanchnic epithelium on the dorsal wall of the pericardial cavity at E11d. At E12d, expression intensity of TGF-βRⅠ and TGF-βRⅡ in myocardium increased to its highest level, and TGF-βRⅡ positive mesenchymal cells in OFT ridges could be detected. After E13d the staining intensity of TGF-βRⅠ and TGF-βRⅡ decreased rapidly， Related Articles | Metrics

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Compatibility of human umbilical vein endothelial cells and surface-modified PHBHHx film EM>in vitro/EM> 2009, 40 (6):  958-962.  doi: 10.3969/j.issn.0529-1356.2009.06.022 Abstract ( )   Objective To evaluate the growing condition of human umbilical vein endothelial cells (HUVECs), which were cultured on the membrane of different component, such as Chinese medicine, before or after the alkali treatment of 3-hydroxybutyrate-co-3-hydroxyhexanoate copolyesters (PHBHHx) and the biocompatibility between PHBHHx flim and endothelial cell. Methods The HUVECs were harvested by Baudine method,and identified by immunohistochemical method.Then the HUVECs of third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. After that, the morphology of HUVECs on different surfaces were observed by scanning electron microscopy, the distribution condition on different membranes were compared by cell-labeling immunofluorescence, and the cell viabilities of all groups were detected by MTT method. Results The HUVECs were successfully separated,and immunohistochemistry staining of FLK-1 and factor Ⅷ was positive.The result of HUVECs culture showed that cells on the material surface growed well, and proliferated significantly. The MTT analysis showed that the PHBHHx film of surface modification and adding some certain proportion of Chinese medicine could promote the growth and proliferation of HUVECs EM>in vitro/EM>,and the cells were thriving, full shape, distribution on the surface by scanning electron microscopy and fluorescence microscopy.Conclusion The PHBHHx film of surface modification and containing certain proportion of Chinese medicine coating had good compatibility of HUVECs, which was favourable to cell growth, adherence and proliferation EM>in vitro/EM>. Related Articles | Metrics

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P>Effect of illumination stress on the expression of GnRH receptor in stomach of SD rat/P> 2009, 40 (6):  963-968.  doi: 10.3969/j.issn.0529-1356.2009.06.023 Abstract ( )   Objective To study the expression of gonadotropin releasing hormone receptor (GnRH receptor) in the stomach of Sprague-Dawley rat exposed to light stress. Methods We established illumination stressed models that the rats were exposed to continuous light(totally 64 rats, 32 rats for experiment and control group, respectively). Then,stomachs were taken from the rats when the rats exposed to continuous light for 1 day, 2 days, 3 days, 4 days, 1 week, 2 weeks, 3 weeks, 4 weeks and their control groups, respectively.The localization and the expression of GnRHR and GnRHR mRNA in stomachs mucous membrane were detected using immunohistochemistry, Western blotting and realtime PCR. Results Immunoreactivity was displayed mostly in the parietal cells in gastric gland. There were not differences of this distribution in all groups of constant illumination and their control. The immunoreactive materials were distributed on membrane and in cytoplasm of all positive cells, but not in nuclei. The level of GnRHR in rats exposed to continuous illumination were higher from one week to four weeks compared with that in control(EM>P/EM><0.05), and the level of GnRHR reached the peak when rats were exposed to constant illumination for two weeks (EM>P/EM><0.01). The quantity of GnRH mRNA in rats exposed to continuous illumination were higher from three weeks to four weeks compared with that in control（EM>P/EM><0.05）. Conclusion The expression of GnRHR in digestive tract was effected by illumination stress.GnRH might regulate digestive function by interaction with GnRHR, suggesting that GnRH may be acted as a hormone, not only responded to normal physiological function of digestive tract but also responded to stre Related Articles | Metrics

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Expression and its significance of Dishevelled2 and Vangl2 proteins in the developing mouse palate 2009, 40 (6):  969-973.  doi: 10.3969/j.issn.0529-1356.2009.06.024 Abstract ( )   Objective To explore the relationship between the expression of Dishevelled2 and Vangl2 proteins and the development of mouse palate. Methods Twenty-four pregnant mice were randomly divided into eight groups, and the mouse embryos were obtained at eight clock of the pregnant day of thirteen(p13d8h), p13d14h，p13d22h，p14d8h，p14d14h，p14d22h，p15d8h and p15d22h respectively, then paraffin sections were made conventionally.The distrubution and dynamic changes of Dishevelled2 and Vangl2 proteins in the embryonic palatal shelves were detected by immunohistochemistry and image analysis. Results It was found that the two kinds of proteins expressed in the epithelium and mesenchyma of the mouse palatal shelves at different development stages. The expression levels of the Dishevelled2,in both of the epithelium and mesenchyme of the palatal shelves, increased first (p13d8h-p13d22h）,then decreased rapidly（p13d22h-p14d14h）, and then increased again（p14d14h-p15d22h）. The expression of Vangl2 protein in the mesenchyma showed a similar trend to that of the Dishevelled2, but there was no obvious regularity in the epithelium. In addition, the expressive levels of both kinds of proteins in the epithelium were significantly higher than those in mesenchyma of the palatal shelves. C Related Articles | Metrics

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Development of early human fetal testes after xenografting into mice 2009, 40 (6):  974-978.  doi: 10.3969/j.issn.0529-1356.2009.06.025 Abstract ( )   Objective To investigate the developmental feasibility of early human fetal testes (<3 months) using xenografting technique and to acquire an accessible donor derivation that is essential for studying human germ cell development. Methods Nine testes from 10-13 weeks aborted fetus were grafted under the back skin of 6 castrated nude mice. Grafts were collected at different time point according to the growth of the donor tissues and the health condition of the recipients. Morphological and histological analyses were performed for the observation of the development of grafted immature testicular tissues. Results The mass of grafts was increased from about 5-7mg to 84.1mg (the biggest). Six of 9 testes were to be in developing. Histological observations showed a significant expansion of seminiferous tubules from (44.26±3.14)μm to (77.69±7.47)μm. Cells dispersedly distributed in seminiferous cords at the time of grafting migrated towards the basal part of seminiferous epithelium. Some germ cells with spermatogonium-like characteristics located on the basement membrane. Sertoli cells were in stages from immature into matured with abundant cytoplasm which were orderly arranged around spermatogonia forming a niche-like structure. Conclusion Testes from early aborted human fetus grafted under the back skin of castrated nude mice showed further development and therefore could be used as an easier accessible donor tissues for the investigation of hu Related Articles | Metrics

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Fine structure and circulation of spleen ellipsoid in EM>Pelodiseus sinensis/EM> 2009, 40 (6):  979-983.  doi: 10.3969/j.issn.0529-1356.2009.06.026 Abstract ( )   Objective To examin and analyze the structure, ultrastructure and the circulation of the spleen ellipsoid in the soft-shelled turtle, EM>Pelodiseus sinensis/EM>.  Methods Twelve turtles were used and observed by light- and electron-microscopy and injection of ink suspension in this study. Results The spleen white pulp was consisted of the periarteriolar lymphatic sheath (PALS) and periellipsoidial lymphatic sheath (PELS). There was no lymphoid nodule in the spleen. Red pulp included splenic cords and splenic sinusoids. The marginal zone was not found in the turtle spleen. When the central arteriole left out of the PALS, it divided into several ellipsoid capillaries which were surround by the PELS. The end of the ellipsoid capillary opened directly to the splenic cord and the blood cells then entered into the splenic sinusoid through the gap between the endothelial cells. The ellipsoid capillary was consisted of simple cuboidal epithelium with an uncompleted basement membrane. The ellipsoid wall was consisted of supporting cells,ellipsoid-associated cells and reticular fibres. Lymphocytes and red cells were always found on the ellipsoid wall. After 40min of the injection of ink suspesion, much carbon particles of ink were restricted on the wall. Conclusion The ellipsoid capillary in the soft-shelled turtle, just like the high endothelial venule, was the important passage of the lymphocytes and blood cells going out and into the lymph tissue. The splenic circulation in the turtle belongs to the opening model. Related Articles | Metrics

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Microsurgical anatomic studies of interforniceal diaterma keyhole approach to interpeduncular cistern 2009, 40 (6):  984-987.  doi: 10.3969/j.issn.0529-1356.2009.06.027 Abstract ( )   Objective To explore the feasibility and operation methods of interforniceal diaterma keyhole approach for operative therapy of apex basilar artery aneurysm. Methods Interforniceal diaterma keyhole approach was designed to interpeduncular cistern with diaterma incision from tuber cinereum to posterior perforated substance and between bilateral mammillary bodies. The simulation operations of interforniceal diaterma keyhole approach were performed in 16 cadaveric heads by assisting with Stryker neuronavigation. Anatomic structures were observed by surgical microscope and measured by Stryker neuronavigation in the keyhole approach operations. Results The operations of interforniceal diaterma keyhole approach could be accomplished successfully in 16 cadaveric heads. The distances from bregma to superior margin of interventricular foramen, superior margin of adhaesio interthalamica, mammillary body, superior margin of aqueduct of midbrain and bifurcation of basilar artery were （68.4±4.6）mm, (66.3±6.0)mm,(86.3±5.3)mm, (82.0±7.6)mm and (91.8±5.0)mm respectively. The length of surgical window of diaterma was （9.5±2.6）mm from tuber cinereum to posterior perforated substance between bilateral mamillary bodies. The apex of basilar artery, P1 and P2 of posterior cerebral artery, superior cerebellar artery, posterior communicating artery and perforating branches from them could be exposed distinctly in interpeduncular cistern. The scope of operative exposure region was front to clivus and dorsum sellae by dissected the Liliequist panniculus, lateral to oculomotor nerve and posterior to interpeduncular fossa. The bifurcation of basilar artery apex was deviation to left in 68.8%. The bilateral posterior cerebral arteries were oblique to the anterolateral in 68.8%. There were 1-4 perforating branches from the apex of basilar artery in the included angle of bilateral posterior cerebral arteries. Conclusion Interforniceal diaterma keyho Related Articles | Metrics

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Microanatomy，histology and CT of arachnoid granulationsin middle cranial fossa 2009, 40 (6):  988-991.  doi: 10.3969/j.issn.0529-1356.2009.06.028 Abstract ( )   Objective To further improve the morphological materials of AGs by microdissection, histology and CT, we observed the arachnoid granulations (AGs) in middle cranial fossa. Methods Thirty-three adult cadaveric heads were used for microsurgical dissection; Histological sections of AG specimens from 3 cadaver heads were examined. Forty patients who had both normal conventional brain CT and computed tomographic venography (CTV) were retrospectively reviewed. Results In middle cranial fossa the AGs occur in the following situations in order of frequency: the middle meningeal sinus, sphenoparietal sinus, lateral foramen rotundum and cavernous sinus. AGs usually show round, oval in shape and irregular in shape. AGs can be divided into individual type and leaflet type under light microscope. The numbers of AGs were observed by microanatomy and CTV were 8.72 and 3.52 respectively. The AGs of cavernous sinus was not localized precisely on CTV. Conclusion Study of the AGs in the middle cranial fossa systematically and comprehensively enriches anatomy and image knowledge. It is helpful in neurosurgical planning and choosing operalion procedure to avoid postoperative complications. Related Articles | Metrics

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Imaging and clinical study of the location relation between vertical facial nerve canal and external acoustic meatus in normal people 2009, 40 (6):  992-996.  doi: 10.3969/j.issn.0529-1356.2009.06.029 Abstract ( )   Objective To observe and measure the anatomical structure of approach of vertical facial nerve canal and put forward the normal measurement range and the location relationship among the vertical segment of facial nerve canal, the posterior wall of external acoustic meatus and the rear edge of external ear，and discuss the relationship and clinical significance between the mastoid gasification and the vertical segment of facial nerve canal. Methods 1. Evaluate the accuracy of CT image of related structure, using spiral CT in scanning four skull specimens, get the horizontal distances of the vertical segment of facial nerve canal to the posterior wall of external acoustic meatus and the rear edge of external in the axial position, and get the sagittal diameter of mastoid (the horizontal distance from the lowest point of external auditory inferior canal to the rear edge of mastoid) and the height (the vertical distance between the lowest points of the external auditory canal wall to the mastoid tip) in the sagittal position. And then saw the skull specimens to measure the distance in the same lay with CT image, and discuss the statistics difference of the distance between the values of CT imaging measurements and the dry entities cranial measurements on hand. 2. Study on people: 118 patients (236 sides) with nonear disorders were randomly selected, among which there were 63 females (126 sides) and 55 males (110 sides). They were subjected to maxillofacial CT scan in the same layer that used above, and the horizontal distances of the facial nerve canal to the posterior wall of external acoustic meatus and the rear edge of external ear were measured. In addition, half of the product of diameter and height of the mastoid was defined as mastoid area, which was used to define the extension of mastoid gasification. Then related analysis and regression analysis were done between the vertical segment of facial nerve canal and the posterior wall of external acoustic meatus, as well as the rear edge of external ear. Results 1.Part of the experiment: There was no significantly different on the indicator values between CT image the entity measurements among the four skull specimens (P>0.05). 2. Study on people: There was no significantly different between left side and right side(EM>P/EM>>0.05), but significantly different between genders(EM>P/EM><0.05). Between mastoid area and the distance from the vertical segment of facial nerve canal to the posterior wall of external acoustic meatus there is inverse correlation, and the relevance has the remarkable significance. However, there was no correlation between mastoid area and the vertical segment of facial nerve canal to the rear edge of external ear. Conclusion There was some relationship between the location of the vertical segment of facial nerve canal and external acoustic meatus. Anatomic position of vertical facial nerve cancal and the posterior wall of external acoustic meatus can be showed clearly. CT and in combination with primitive axial images may provide reliable evidence for the diagnosis facial nerve dieases and the choice of ear surgery route. Related Articles | Metrics

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MRI measurement of the epicondyles of the distal femur 2009, 40 (6):  997-1000.  doi: 10.3969/j.issn.0529-1356.2009.06.030 Abstract ( )   Objective Measure the epicondyles of the distal femur on magnetic resonance image(MRI), in order to locate presicely the surgical transepicondylar axis(STEA) in total knee arthroplasty(TKA) and to provide theoretical basis for the designing of the size of the femoral component. Methods Totally 78 normal knees of Chinese individuals were studied. The images of coronal, sagittal and traverse sections of the knee were obtained by magnetic resonance image. Measurement included the width of the STEA, distance between the epicondyles and the joint line, anterior posterior width of the medial and lateral epicondyles, posterior condyle angle. Results The width of the STEA averaged(79.55±4.90)mm in males, and femles(71.18±4.22)mm. The distance from the epicondyles to the joint line was correlated with the width of the STEA, so was the anterior posterior width of epicondyles. PCA averaged(4.22±2.07)°. Conclusion The size of the epicondyles in Chinese is significantly smaller than that of the Westerns. The ratio between anterior posterior width of the medial epicondyle and the width of the STEA is 0.84, and is 0.87 between anterior posterior width of the lateral epicondyle and the width of the STEA. The distance from the epicondyles to the joint line is helpful to locate the STEA. The reliability is poor to lo Related Articles | Metrics

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Sex differences in the length ratios of metapodials inEM>Macaca mulatta/EM> from the Taihang mountains 2009, 40 (6):  1001-1004.  doi: 10.3969/j.issn.0529-1356.2009.06.031 Abstract ( )   Objective Sex difference of the length ratios of metacarpals and metatarsals in Macaca mulatta from the Taihang mountains was studied in our laboratory. Methods The lengths of 27 metacarpals (10 males, 17 females) and 30 metatarsals(12 males, 18 females) were measured from the skeletons of 30 adult EM>Macaca mulatta/EM>. Length ratios were constructed for all possible pairings of the five bones in each individual hand and foot. EM>One-Way ANOVA/EM>  adopting SPSS13.0 for windows was used to study the sex differences of length ratios of metacarpals and metatarsals. Results For EM>Macaca mulatta/EM>, several of these lengths ratios exhibited substantial differences between the sexes. The metacarpal(Mc) length ratios showing the largest sex differences were 2Mc∶5Mc and 4Mc∶5Mc in both hands (EM>P/EM><0.01), and the metatarsal(Mt) length ratios showing the largest sex difference was 1Mt:3Mt in both feet (EM>P/EM><0.05). Conclusion The sex differences of metacarpals and metatarsals remained when specimens of similar size were compared. It showed that body size was not the basis for these sex differences. Various facts suggested that the sex difference of length ratios in primate metapodials was associated with sex hormones exposure, possibly Related Articles | Metrics

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Distributions of purinergic P2XSUB>3/SUB> receptor immunoreactive afferent fibers in the rat pharyngeal mucosa 2009, 40 (6):  1005-1007.  doi: 10.3969/j.issn.0529-1356.2009.06.032 Abstract ( )   Objective To study the distribution of purinergic P2XSUB>3/SUB> receptor immunoreactive afferent fibers in the rat pharyngeal mucosa and provide morphological evidence for exploring the role of ATP in the signal transduction of sensory stimuli. Methods Twelve adult Wistar rats were used and immunofluorescenthistochemical double-labeling technique combined laser confocal scanning microscope was applied in the present study. Results 1.P2XSUB>3/SUB> immunoreactive fibers were observed on sections from all parts of pharyngeal mucosa. Two types of positive fibers were found. One was free nerve fibers covered with many varicosities. Another one ramified in mucosa and showed complex arborization endings. Nerve plexus in mucosa were formed by P2XSUB>3/SUB> immunoreactive fiber ramifications. 2.Most calcitonin generelated peptide (CGRP) positive fibers intermingled with P2XSUB>3/SUB> immunoreactive fiber arborizations. A few P2XSUB>3/SUB> immunoreactive fibers were covered with many varicosities co-localized with CGRP. 3.In petrosal ganglion, most neurons were stained with P2XSUB>3/SUB> or CGRP immunoreactivity and small number of P2XSUB>3/SUB> immunoreactive neurons costained with CGRP. Conclusion These results indicated that the different types of afferent fibers in rat pharyngeal mucosa expressed purinergic P2XSUB>3/SUB> receptor immunoreactivity and ATP might be related to nociceptive or physiological signals transduction in rat pharyngeal mucosa. Related Articles | Metrics

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Anatomical study of the femoral and tibial insertions of the anterolateral and posteromedial bundles of human posterior cruciate ligament 2009, 40 (6):  1008-1011.  doi: 10.3969/j.issn.0529-1356.2009.06.033 Abstract ( )   Objective To provide an anatomic evidence for the double-bundle posterior cruciate ligament (PCL) reconstruction, the sizes and locations of the attachments of the PCL to the tibia and the femur were measured. Methods We studied 30 cadaveric knees. PCLs were divided into anterolateral and posteromedial bundles to the insertion footprint, and those locations were measured and described. Results The distances from the center of the femoral insertions of the anterolateral and posteromedial bundles to the anterior margin of the medial femoral condyle were (8.52±1.81)mm and (11.63±1.81)mm. The vertical distances from the center of the femoral insertions of the double-bundle to the intercondylar roof were (4.67±0.55)mm and (10.32±1.23) mm. The vertical distances from the tibial insertion of the center of the double-bundle to the plane of the tibial articular surface were (8.43±1.21)mm and (14.52±2.31)mm. The distances from the medial margin of the articular cartilage of the tibial plateau to the center of the tibial insertions of double-bundle were (47.44±6.23)mm and (45.95±6.32)mm. The areas of the insertions of the anterolateral and posteromedial bundles on the femur were (107.12±15.25)mmSUP>2/SUP> and (65.35±10.27)mmSUP>2/SUP>. The areas of the insertions of the double-bundle on the tibia were (50.07±11.33)mmSUP>2/SUP> and (51.08±10.22 Related Articles | Metrics