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Objective: To improve the survival status of wild Rheum tanguticum, which is threatened to meet the market demand, Illumina high-throughput sequencing technologies is used to bring new directions for relevant research. Methods The complete chloroplast genome of Rheum tanguticum was constructed with Illumina high-throughput sequencing technologies in this paper. Results: The genome was 161 054 bp in length, and exhibited a typical quadripartite structure of the large (LSC, 86 441 bp) and small (SSC, 12 745 bp) single-copy regions, separated by a pair of inverted repeats (IRs, 30 934 bp each). The chloroplast genome contained 132 genes, including 88 protein-coding genes, 36 transfer RNA genes, eight ribosomal RNA genes and two pseudogenes, 19 genes located in each IR area. Conclusion: The phylogenetic tree constructed with 11 sequences of seven Polygonaceae species and four other family species demonstrated a close relationship between R. tanguticum and R. palmatum in Polygonaceae, which was coincided with their morphological similarity, in addition, there were certain SNP sites in rpl32 and other genes, which provided a new basis for the effective identification of related species.
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<p><b>OBJECTIVE</b>To detect the plasma levels of mannan?binding lectin (MBL) and MBL?associated serine protease?2 (MASP-2) in patients with hepatocellular carcinoma (HCC) and explore their role in the tumorigenesis and progression of HCC.</p><p><b>METHODS</b>The plasma levels of MBL and MASP?2 were detected by enzyme?linked immunosorbent assay in 64 HCC patients and 30 healthy control subjects. The correlation of MBL and MASP?2 with the clinical parameters of HCC patients were analyzed.</p><p><b>RESULTS</b>The plasma levels of MBL (P=0.014) and MASP?2 (P=0.002) were significantly higher in HCC patients than in the healthy controls, but the MBL?to?MASP?2 ratio did not differ significantly between the two groups. In HCC patients, plasma MBL level was positively correlated with vascular invasion (r=0.253, P=0.047) and total bilirubin level (r=0.283, P=0.024). The plasma level of MASP?2 was positively correlated with TNM stage (r=0.276, P=0.027) and negatively correlated with plasma albumin level (r=0.?0.317, P=0.015). ROC curve analysis revealed an area under curve of 0.665 for MBL (P=0.010) and 0.694 for MASP?2 (P=0.003). The sensitivities of MBL and MASP?2 were 50% and 89.1% in the diagnosis of HCC, respectively.</p><p><b>CONCLUSION</b>MBL and MASP?2 are associated with the inflammatory state and disease progression in patients with HCC.</p>
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OBJECTIVE: To explore the mechanism of bone marrow mesenchymal stem cells( BMSCs) in alleviating pulmonary alveolitis in mice exposed to silica dust. METHODS: Five specific pathogen free healthy male C57 BL /6 mice were used to isolate BMSCs using bone marrow adherent method. The poly-potent differentiation ability of BMSCs were identified by 3 differentiation-inducing experiments. Forty-five mice of similar background were randomly divided into 3groups: control group,silica group and BMSCs transplantation group. The mice of the control group were given 20. 0 μL of0. 90% sodium chloride solution by one time intratracheal injection. The mice of silica group and BMSCs transplantation group were first received 20. 0 μL( 250 g / L mass concentration) of silica dust suspension by one time intratracheal injection; followed by 500. 0 μL of 0. 90% sodium chloride solution or 500. 0 μL of BMSCs suspension( cell density 1 ×109/ L) by tail vein infusion 6 hours later. Mice were euthanized on the 3rd day of the experiment. Lung functional coefficient and pathologic changes in the lung were examined. The level of cytokines in bronchoalveolar lavage fluid( BALF) was detected by enzyme linked immunosorbent assay. Wright-Giemsa staining was used for staining cells in BALF for counting. Flow cytometry( FCM) was used to measure the percentage of macrophages of BALF in the mice. RESULTS: BMSCs were successfully induced to differentiate into osteogenic,adipogenic and chondrogenic cells and developed into osteoblast,adipogenic cells and chondroblast. On the 3rd day of the experiment,the mice in silica group showed histopathological changes similar to pulmonary alveolitis; while there was no obvious inflammatory change observed in the BMSCs transplantation group,and the structure of lung tissue appeared normal. The lung coefficient of the silica group was higher than that of the control group( P < 0. 05); the lung coefficient of BMSCs transplantation group was lower than that of the silica group( P < 0. 05),but it showed no significant difference when compared to the control group( P > 0. 05). The interleukin( IL)-1β,IL-6 and chemokine ligand 3 levels in BALF in the silica group were higher than those of the control group( P < 0. 05),and the above 3 indices in the BMSCs transplantation group regaining the level of the control group( P > 0. 05) were lower than those of the silica group( P < 0. 05). The level of tumor necrosis factor-α in BALF in silica group and BMSCs transplantation group were higher than that of the control group( P < 0. 05),but there was no significant difference between silica group and BMSCs transplantation group( P > 0. 05). The level of IL-10 in BALF showed no significant difference in these 3 groups( P > 0. 05). Wright-Giemsa staining results showed that the number of total cells and macrophages in BALF in the silica group was higher than that of the control group( P < 0. 05),and the above cell number of BMSCs transplantation was lower than that of silica group( P < 0. 05),but it showed no significant difference when compared to the control group( P > 0. 05). The FCM result showed that the percentage of macrophages was in accordance with that of the Wright-Giemsa staining. CONCLUSION: The BMSCs can alleviate pulmonary alveolitis in the mice exposed to silica dust by inhibiting the amounts and activity of alveolar macrophages and down-regulating the expression of IL-1β and IL-6 in BALF.
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OBJECTIVE: To study the effects of bone marrow mesenchymal stem cell( BMSC) transplantation on early stage of inflammation in mice exposed to silica dust. METHODS: Specific pathogen free healthy male C57 BL /6 mice were used.Five mice were used to isolate BMSCs using bone marrow adherent method. Thirty mice were randomly divided into 3groups: control group,silica group and BMSCs transplantation group. The mice of the control group were given 20. 0 μL of0. 90% sodium chloride solution by one time intratracheal injection. The mice of silica group and the BMSCs transplantation group first received 20. 0 μL( 250 g / L mass concentration) of silicosis dust suspension by one time intratracheal injection; followed by 500. 0 μL of 0. 90% sodium chloride solution in the silica group,and 500. 0 μL of BMSCs suspension( cell density 1 × 109/ L) by tail vein infusion in the BMSCs transplantation group 6 hours later. Mice were euthanized on the 7th day of the experiments. The histopathology changes in lung tissues were examined. The serum levels of interleukin( IL)-1β, IL-6 and IL-10 were detected by enzyme linked immunosorbent assay. Real-time quantitative polymerase chain reaction was used to detect the mRNA relative expression levels of above cytokines in the lung tissues. RESULTS: The positive rates of BMSCs surface molecules cluster differentiation( CD) 29,CD34,CD90,CD105 and CD106 were 67. 70%,0. 12%,39. 00%,37. 10% and 20. 10%,respectively. Histopathology examination showed the thickened alveolar walls,broadening alveolar septum and the damaged alveolar structure in silica group. In the BMSCs transplantation group,there was no obvious damage found in the lung tissue. There was no change in the alveolar cavity and alveolar structure was complete. The IL-1β and IL-6 levels in serum and mRNA relative expression of IL-1β and IL-6in lung tissue in the silica group were higher than those of the control group and BMSCs transplantation group( P < 0. 05).The IL-1β level in serum and mRNA relative expression of IL-1β in lung tissue in the BMSCs group were higher than those of the control group( P < 0. 05). The IL-10 level in serum and mRNA relative expression of IL-10 in lung tissue in all groups showed no statistical difference( P > 0. 05). CONCLUSION: The BMSCs can alleviate pulmonary inflammatory damage at early stage by down-regulating the expression of proinflammatory factors of IL-1β and IL-6.
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Objective: To investigate the chemical constituents of monoterpenes in the fruits of Gardenia jasminoides. Methods: Various column chromatographies were used in the isolation and purification, and the physicochemical constant determination and spectral analysis were adopted to identify the chemical structures of monoterpenes. Results: Twelve monoterpenes were isolated from G. jasminoides, such as jasminoside B (1), jasminoside G (2), jasminodiol (3), crocusatin-C (4), (7S)-6-(hydroxymethyl)-1, 1, 5-trimethylcyclohex-3-enone (5), bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside (6), (10R, 11R)-gardendiol (7), (10S, 11S)-gardendiol (8), (5S, 9S)-gardenate A (9), (5R, 9R)-gardenate A (10), jasminoside E (11), and 5, 6-dihydroxymethyl-1, 1-dimethylcyclohex-4-enone (12). Conclusion: Compounds 5, 6, 8, 10, and 12 are first isolated from this plant.
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<p><b>OBJECTIVE</b>To observe the maximal heart rate changes, atrioventricular (A-V) conduction block and atrial fibrillation (AF) inducibility in dogs with vagosympathetic trunk exposed to electromagnetic fields (EMFs).</p><p><b>METHODS</b>The vagosympathetic trunk of adult dogs was separated and exposed to EMFs 0.043 kHz (2.87 microG, n = 5) and to EMFs 2 kHz (0.34 microG, n = 6) for two to three hours. Simultaneously, the vagosympathetic trunk was stimulated with 20 Hz frequency and 1 - 8 V intensity for 0.1 ms. Heart rate, presence of A-V conduction block and AF inducibility were determined.</p><p><b>RESULTS</b>After 5-minutes exposure to EMFs 0.043 kHz (2.87 microG), the maximal heart rate decreased 29%, the voltage applied to vagosympathetic trunk required to induce A-V conduction block decreased by 60% in experimental group versus 5% increase in control group. This effect lasted 2 to 3 hours. While vagosympathetic trunk exposure to EMFs 2 kHz (0.34 microG) was associated with significant increase in the incidence of atrial premature beats, atrial tachycardia and AF, these effects could be blocked by propranolol and atropine.</p><p><b>CONCLUSIONS</b>Our results showed that 0.043 kHz (2.87 microG) EMFs exposure might reduce while 2 kHz (0.34 microG) EMFs exposure might increase AF inducibility. Our study thus suggested autonomic nervous system of dogs could be affected by EMFs exposure and 0.043 kHz (2.87 microG) EMFs exposure might be a novel option for AF prevention.</p>
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Animals , Dogs , Atrial Fibrillation , Heart Rate , Magnetics , Vagus NerveABSTRACT
<p><b>OBJECTIVE</b>To express the carbohydrate recognition domain (CRD) of Balb/C mouse mannan binding lectin A (MBL-A) in E.coli.</p><p><b>METHODS</b>The target gene fragment was obtained by PCR from the plasmid pmMBL-A harboring mouse MBL-A gene. The PCR product was recombined with the prokaryotic expression vector pET-41a(+) and the resulting recombinant plasmid was identified by PCR, restriction analysis and sequencing before transformation into E.coli BL21(DE3) cell for expression of the target protein. After washing and renaturation, the protein was purified on GST-Tag purification resins and analyzed by SDS-PAGE, Western blotting and enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>A DNA fragment of about 450 bp was amplified by PCR and the recombinant plasmid pET41a-mMBL-A-CRD was constructed by linking the fragment with pET41a(+) vector. The result of restriction enzyme analysis and sequencing of the selected clones were consistent with those by computer analysis. The recombinant vector was expressed in E.coli BL21(DE3), and the expressed protein existed mainly as inclusion bodies, whose relative molecular mass was about 47,000 by SDS-PAGE analysis. After washing, renaturation and purification, the purity of recombinant protein was about 90%. Western blotting suggested immunoreactivity of the purified protein with anti-GST antibody, and its sugar binding activity was verified by ELISA.</p><p><b>CONCLUSION</b>We have successfully obtained mouse MBL-A CRD protein, which provides the base for further functional study of the MBL-A molecule.</p>
Subject(s)
Animals , Mice , Carbohydrates , Chemistry , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Inclusion Bodies , Metabolism , Mannose-Binding Lectin , Chemistry , Genetics , Mice, Inbred BALB C , Recombinant Fusion Proteins , Chemistry , GeneticsABSTRACT
Objective To explore the relationship of dynamic changes between the iodine nutritional condition and growth of children born before and after the implementation of universal iodine.Methods By means of sampling investigation,urinary iodine and goiter,height,weight,intelligence quotient(IQ)were comprehensively evahated in children aged 7～14 years old born before and after the implementation of iodine in 10 counties and 2 cities of Qiannan.Results The median of urinary iodine of children Was 91.88μg/L in 1984,133.33μg/L in 1994,316.00μg/L in 2006;goiter rate was 21.40%(122/570)in 1984,12.86%(107/832)in 1994,5.28%(45/851)in 2006;the average IQ was 89.18±4.10 in 1994,94.26±0.94 in 2006;children in each age group in 2006 has a higher height and a heavier weight compared with those in 1984.Conclusion The physical development and intelligence of children in the areas of iodine deficiency have improved along with the correction of iodine deficiency.
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Objective To investingate the iodine-nutritional status of the rural children form 8 to 10 years old after salt iodzation had been implemented in Qiannan State.Methods The size of thyroid gland waft measured by palpation in 421 rural children aged 8～10 years old in Qiannan State.Calorimetric eerie-arsenic assay and vitriolic ammonium assimilmion were used for testing urinary iodine,while Rsvens test was used to test the intelligent quotient (IQ).Results The rate of goiter was 5.0%(21/421).The median of urinary iodine Was 331.2μg/L The average IQ was 93.32±18.68 of the children aged 8～10.The IQ of children with different iodine-nutritional level was significantly different(P＜0.05).Conclusions The children's iodine nutrition in Qiannan State has been improved after the salt adds the iodine.Their intelligence level is quite normally.Iodine dificiency disease has been eliminated according to the national standard in terms of 8～10 year old child ufinaty iodine level and goiter rate.
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<p><b>OBJECTIVE</b>To obtain highly purified tetanus toxin fragment C (TTC) with good immunogenicity.</p><p><b>METHODS</b>The gene fragment encoding TTC was amplified from Clostridium tetani plasmid DNA by PCR, inserted into the vector pET43.1a (+) and expressed in E. coli BL21(DE3)plysS. After purification using Ni2+-chelate affinity chromatography, the expressed fusion protein was digested by thrombin and the resultant TTC protein was purified with Ni2+-chelate affinity chromatography followed by identification with SDS-PAGE and Western blotting. The purifed TTC protein was then used to immunize mice to test its immunogenecity.</p><p><b>RESULTS</b>The 1373-bp gene fragment encoding TTC was obtained, and the constructed recombinant expression vector pET43.1a (+)-TTC was successfully expressed in E. coli BL21(DE3)plysS. SDS-PAGE identified a recombinant fusion protein with relative molecular mass (Mr) of 117 000, which accounted for 22% of the total bacterial protein. The TTC protein with Mr of 50 000 was obtained after purification of the thrombin digestion products of the fusion protein, with a purity reaching 95.5%. Both the fusion protein and TTC protein could be recognized by anti-tetanus toxin antibody as shown by Western blotting. The titer of the anti-serum from mice immunized with the TTC protein was 1:25 600, and the anti-serum could specifically bind to tetanus toxin.</p><p><b>CONCLUSION</b>Highly purified and immunogenetic TTC protein has been successfully obtained, which provides a good model antigen for studying antigen presentation and immune responses in vivo.</p>
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Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Peptide Fragments , Genetics , Allergy and Immunology , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Tetanus Toxin , Genetics , Allergy and Immunology , Tetanus Toxoid , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To clone the gene coding for the peptidoglycan recognition protein (PGRP) domain (PGRPd) of mouse long PGRP (mPGRP-L) and express the protein in E. coli.</p><p><b>METHODS</b>The cDNA fragment encoding PGRPd of mPGRP-L was obtained by RT-PCR from the total RNA of Balb/C mouse liver cells and cloned into pUCm-T vector. The recombinant plasmid were identified by PCR, restriction endonucleases and sequence analysis. The PGRPd gene fragment was amplified by PCR from the recombinant plasmid, inserted into pQE-30 vector and transformed into E. coli strain M15, and the expressed PGRPd protein was purified.</p><p><b>RESULTS</b>A cDNA fragment of about 500 bp was amplified by RT-PCR and the recombinant plasmid, pmPGRPd, was constructed by linking the fragment to pUCm-T vector. The results of restriction mapping of the recombinant vector were consistent with those of computer analyses. Sequence analysis showed that the cloned gene fragment (518 bp) had identical sequence with the gene encoding PGRPd of mPGRP-L gene in GenBank. The recombinant expression vector pQE-PGRPd was constructed and expressed in E. coli M15. SDS-PAGE showed that the expressed product existed mainly in the lysate supernatant as a soluble protein with relative molecular mass of 29 kD.</p><p><b>CONCLUSION</b>The PGRPd cDNA of mPGRP-L has been successfully cloned and expressed in E. coli, which provides the basis for further study of PGRP molecule.</p>
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Animals , Female , Mice , Base Sequence , Carrier Proteins , Genetics , Cloning, Molecular , DNA, Complementary , Genetics , Escherichia coli , Genetics , Metabolism , Liver , Chemistry , Mice, Inbred BALB C , Molecular Sequence Data , Plasmids , Genetics , Prokaryotic Cells , Metabolism , Protein Structure, Tertiary , Genetics , Recombinant Proteins , GeneticsABSTRACT
Objective To study the correlation of expression of N-myc downstream regulated gene 1(NDRG1) with invasion of breast invasive duct carcinoma and lymph node metastasis. Methods A total of 71 specimens including 26 case of primary breast invasive duct carcinoma with lymphnode metastasis,45 case of nonmetastasis breast invasive duct carcinoma were observed.NDRG1 was detected by immunohistochemistry in formalin-fixed and paraffin-embedded sections.At the same time,the correlations of NDRG1 with E-cad,MMP2,MMP9,and TIMP2 were investigated. Results The expression of NDRG1 in breast cancer with metastasis of lymph nodes(9/26) was lower than that of non-metastasis of lymph nodes(32/45)(P
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<p><b>OBJECTIVE</b>To investigate the essentials of operation and the postoperative effect of right hemicolonic carcinoma with anterograde clearance of lymph nodes.</p><p><b>METHODS</b>One hundred and thirty-five patients with right hemicolonic infiltrated carcinoma, who were eligible for radical excision (D(3)), were divided into 2 groups. Among them, the anterograde clearance of lymph nodes was performed on 56 cases; the retrograde clearance was performed on 79 cases. Both groups showed no significant difference in age, sex, Dukes' staging and pathological type.</p><p><b>RESULTS</b>The average time of operation: the anterograde group was (180 +/- 40) min; the retrograde group was (180 +/- 20) min. The average amount of bleeding: the anterograde group was (200 +/- 80) ml; the retrograde group was (200 +/- 30) ml. The cleared number of lymph nodes: the anterograde group were 6.3 +/- 4.2, 2.6 +/- 3.1, 1.5 +/- 2.3 in paracolon, middle and radicel of vasorum respectively, the total number was 11.4 +/- 8.6; the retrograde group were 6.4 +/- 2.2, 2.8 +/- 2.1, 1.1 +/- 1.1 respectively, the total number was 10.8 +/- 5.6 (P > 0.05). The postoperative metastasis to liver: the anterograde group was 8 cases (13.9%); the retrograde group was 21 cases (26.6%, P < 0.05). The 5-year survival rate: the anterograde group was 72.8% (41/56); the retrograde group was 65.5% (52/79) (P < 0.05).</p><p><b>CONCLUSIONS</b>The operative technique of the excision was little difficulty and complexity, and it could fit well with the requirement of non touch isolation, and act to cut down the postoperative metastasis to liver and to elevate 5-year survival rate.</p>
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Adult , Female , Humans , Male , Middle Aged , Colonic Neoplasms , Mortality , General Surgery , Follow-Up Studies , Lymph Node Excision , Methods , Retrospective Studies , Survival Rate , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To consider the relationship to survival rate and quality of life with pararectal lymphadenectomy for lower carcinoma of rectum.</p><p><b>METHODS</b>The radical operation was performed on 780 cases of progressive cancer located at peritoneal reflection or below it, Among them, 352 cases only cleared in abdominal cavity, 428 cases coupled with extra-peritoneal histopathological type.</p><p><b>RESULTS</b>Urinary function injured, the group cleared in abdominal cavity was 12 cases, accounted for 3.6%; the group coupled with extra-peritoneal clearance was 225 cases, for 52.5% (P < 0.01). Sexual function damaged (only for male), the abdominal cavity group was 23 cases, for 12.6% (23/185); the coupled group was 127 cases, for 53.4% (127/238), (P < 0.01). Local relapse rate, the abdominal cavity group was 15.8% (56/352); the coupled group was 8.6% (41/428), (P < 0.05). 5-year survival rate, the abdominal cavity group was 52.2%; the coupled group was 58.5% (P < 0.05).</p><p><b>CONCLUSION</b>By contrast, although abdominal cavity coupled with extraperitoneal lymphadenectomy acted to cut down local relapse and to elevate 5-year survival rate, the postoperative quality of life appeared to be seriously affected.</p>
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Female , Humans , Male , Lymph Node Excision , Methods , Proctocolectomy, Restorative , Quality of Life , Rectal Neoplasms , Mortality , Pathology , General Surgery , Retrospective Studies , Survival RateABSTRACT
<p><b>OBJECTIVES</b>To evaluate the effect and the indication of local resection for rectal cancer below the peritoneal reflection.</p><p><b>METHODS</b>One hundred and thirty-four patients with early rectal carcinoma whose masses were >or= 5 cm or <or= 7 cm apart from the anus border, eminence or surface shape, 1 - 3 cm in the diameter, and could be shook up and down or sideways at basal part. Trans-sacral local resection was performed in all patients.</p><p><b>RESULTS</b>No recurrence appeared in 47 patients with intra-mucosal carcinoma (M-Ca). In 87 patients with submucosal carcinoma (SM-Ca), 3 patients were positive for the cutting end, 8 showed vessel infiltration, and 5 showed low-differentiated adenocarcinoma. These patients had to accept mile's operation, but one patient had local recurrence and 2 patients had liver metastases. Two patients had local recurrence and 8 had metastasis of the liver and other sites. The total 5-year survival rate was 94%.</p><p><b>CONCLUSIONS</b>Trans-sacral local resection is an optimal operation since the operative range is small and artificial anus is avoided. However, it is only suitable for early rectal carcinoma. When vessel infiltration or low differentiation or positive cutting end are confirmed, radical operation should be added.</p>
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Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Follow-Up Studies , Neoplasm Invasiveness , Neoplasm Recurrence, Local , Neoplasm Staging , Rectal Neoplasms , Pathology , General Surgery , Rectum , Pathology , General Surgery , Survival Analysis , Treatment OutcomeABSTRACT
Objective To investigate the survival rate, voiding, sexual function after nerve-sparing surgery with extended systematic extraperitoneal lateral lymphadenectomy in lower rectal cancer. Methods Three hundreds and ninety-two cases with advanced lower rectal cancer who underwent nerve-sparing surgery with radical dissection from 1996 to 2000 were reviewed. Among them,173 cases only cleared in abdominal cavity,219 cases coupled with extraperitoneal lateral lymphadenectomy. Results The metastatic rate of lateral lymph node was 17.8%(39/219),the rate of non-modal foci of metastatic disease in lateral out of the abdominal cavity was 5.9%(13/219).Local recurrence rate, the abdominal cavity group was 16.2%(28/173); the coupled group was 9.6%(21/219), P